»«. U1 ■-S*, ». '•V-.. i.-«x>- ' .^^t. .„,5 "--u.^- V ■'> ■< ■;'f^^- > _;■) ^«.^ " m-^. t-K ■'-mm irjL^... -V-^ ■^:r4 ^t.i'& Mxm BIOCHEMICAL BULLETIN ISSUED QUARTERLY BY THE COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION PRESS OF THE NEW ERA PRINTING COMPANY LANCASTER, PA. BIOCHEMICAL BULLETIN VOLUME III Nos. 9-12 1913-1914 WITH THREE PORTRAITS AND SEVEN PLATES NEW YORK Columbia University Biochemical Association 1914 Entered as second-class matter in tbe Post Office at Lancaster, Pa. /^ h^ EDITORIAL COMMITTEE: Herman M. Adler, John S. Adriance, David Alperin, Carl L. Aisberg, D. B. Armstrong, George Baehr, Louise C. Ball, Charles W. Ballard, Louis Baumann, George D. Beal, S. R. Benedict, William N. Berg, Josephine T. Berry, Robert Bersohn, Isabel Bevier, Louis E. Bisch, A. Richard Bliss, Charles F. Bolduan, Samuel Bookman, Sidney Born, O. C. Bowes, William B. Boyd, J. Bronfenbrenner, Jean Broadhurst, Leo Buerger, Gertrude Burlingham, J. G. M. Bullowa, R. Burton-Opitz, A. M. Buswell, R. P. Calvert, A, T. Cameron, Herbert S. Carter, Arthur F. Chace, Ella H. Clark, Ernest D. Clark, Alfred E. Cohn, Burrill B, Crohn, Louis J. Curtman, William D. Cutter, C. A. Darling, William Darrach, Norman E. Ditman, Eugene F. DuBois, James G. Dwyer, Walter H. Eddy, Gustave Egloff, A. D. Emmett, Allan C. Eustis, Benjamin G. Feinberg, Ruth S. Finch, Harry L. Fisher, Mabel P. FitzGerald, Nellis B. Foster, Fred D. Fromme, C. Stuart Gager, Mary C. de Garmo, Helen Gavin, Mary E. Gearing, George A. Geiger, William J. Gies, Samuel Gitlow, A. J. Goldfarb, H. D. Goodale, H. B. Goodrich, F. G. Goodridge, Ross A. Gortner, Mark J. Gottlieb, Isidor Greenwald, James C. Greenway, Louise H. Gregory, Abraham Gross, B. C. Gruenberg, Marston L. Hamlin, Frederic M, Hanes, R, F. Hare, Tula L. Harkey, T. Stuart Hart, E. Newton Harvey, P. B. Hawk, Harold M. Hays, M. Heidelberger, Joseph S. Hepburn, Alfred F. Hess. L. J. Hirschleifer, Benjamin Horowitz, Homer D. House, Paul E. Howe. Louis Hussakof, Roscoe R. Hyde, Henry H. Janeway, Max Kahn, John L. Kantor, Edward C. Kendall, J. E. Kirkwood, Israel J. Kligler, Arthur Knudson, Mathilde Koch, Walter M. Kraus, Alfred H. Kropff, Marguerite T, Lee, Victor E. Levine, Charles C. Lieb, •> B. E. Livingston, Leon Loewe, Alfred P. Lothrop, Daniel R. Lucas, Wm. H, McCastline, Mary G. McCormick, Louise McDanell, Grace MacLeod, C. A. Mathewson, H. A. Mattill, Clarence E. May, Gustave M. Meyer, E. G. Miller, Jr. Sergius Morgulis, Max Morse, H. O. Mosenthal, Hermann J. Muller, Archibald E. Olpp, iv B. S. Oppenheimer, Raymond C. Osburn, Reuben Ottenberg, A. M. Pappenheimer, Olive G. Patterson, W. A. Perlzweig, W. H. Petersen, Louis Pine, E. R. Posner, P. W. Punnett, Jessie Moore Rahe, A. N. Richards, Anna E. Richardson, Winifred J. Robinson, Anton R. Rose, Jacob Rosenbloom, William Salant, W. S. Schley, Oscar M. Schloss, H. von W. Schulte, Fred W. Schwartz, C. A. Schwarze, Emily C. Seaman, Fred J. Seaver, A. D. Selby, A. Franklin Shull, Clayton S. Smith, Edward A. Spitzka, Matthew Steel, Ralph G. Stillman, Charles R. Stockard, Edward C. Stone, Mary E. Sweeny, Arthur W. Thomas, Helen B. Thompson, F. T. Van Buren, Eliz. G. Van Hörne, Philip Van Ingen, Charles H. Vosburgh, Hardolph Wasteneys, Edwin D. Watkins, William Weinberger, F. S. Weingarten, J. W. Weinstein, Charles Weisman, William H. Welker, C. A. Wells, Harry Wessler, H. L. White, Geo. H. Whiteford, David D. Whitney, Ethel Wickwire, Herbert B. Wilcox, Guy W. Wilson, Louis E. Wise, William H. Woglom, L. L. Woodruff, H. E. Woodward, R. M. Yergason, Hans Zinsser. MEMBERS OF THE COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION Honorary Members PROF. R. H. CHITTENDEN, First Director of the Columbia University De- partment of Biological (Physiological) Chemistry and Director of the Sheffield Scientific School of Yale University ♦PROF. HUGO KRONECKER, Director of the Physiological Institute, Uni- versity of Bern, Switzerland. (Died, June 6, 1914-) PROF. SAMUEL W. LAMBERT, Dean of the Columbia University School of Mediane DR. JACQUES LOEB, Member of the Rockef eller Institute for Medical Re- search and Head of the Department of Experimental Biology PROF. LAFAYETTE B. MENDEL, Professor of Physiological Chemistry, Sheffield Scientific School, Yale University PROF. ALEXANDER SMITH, Head of the Department of Chemistry, Co- lumbia University Corresponding Members PROF. EMIL ABDERHALDEN, University of Halle, Gennany PROF. LEON ASHER, University of Bern, Switzerland PROF. FILIPPO BOTTAZZI, University of Naples, Italy PROF. ROBERT B. GIBSON, University of the Philippines, P. I. PROF. VLADIMIR S. GULEVIC, University of Moscow, Russia PROF. W. D. HALLIBURTON, King's College, London PROF. S. G. HEDIN, University of Upsala, Sweden PROF. FREDERICO LANDOLPH, University of La Plata, Argentina PROF. A. B. MACALLUM, University of Toronto, Canada PROF. D. McCAY, Medical College, Calcutta, India PROF. W. A. OSBORNE, University of Melbourne, Austraiia. PROF. C. A. PEKELHARING, University of Utrecht, Holland PROF. S. P. L. SÖRENSEN, Carlsberg Laboratory, Copenhagen, Denmark Members Resident in New York City Brooklyn Botanic Garden. — C. Stuart Gager. College of the City of New York. — ^Wm. B. Boyd, Louis J. Curtman, Benj. G. Feinberg, A. J. Goldfarb. Columbia University: Departments. — Agriciilture : O. C. Bowes; Anat- omy: Alfred J. Brown, H. von W. Schulte; Bacteriology: James G. Dwyer, Reuben Ottenberg, Hans Zinsser; Biological Chemistry: Walter H. Eddy, William J. Gies, F. G. Goodridge, Tula L. Harkey, Paul E. Howe, Arthur Knudson, Victor E. Levine, Alfred P. Lothrop, Sergius Morgulis, H. O. Mosen- thal, W. A. Perlzweig, Emily C. Seaman, Chris Seifert, Charles Weisman, Ethel Wickwire; Botany: E. R. Ahenburg, C. A. Darling; Cancer Research: W. H. Woglom; Chemistry: A. M. Buswell, R. P. Calvert, Gustave Egloflf, H. L. Fisher, P. W. Punnett, A. W. Thomas; Clinical Pathology: Edward Cussler, Peter Irving, Arthur W. Swann; Diseases of Children: Chas. Hendee Smith, Herbert B. Wilcox; Gynecology: Wilbur Ward; Mediane: T. Stuart Hart, I. Ogden Woodruff; Pathology: B. S. Oppenheimer, Alwin M. Pappen- heimer; Pharmacology: Charles C. Lieb; Physiology: Russell Burton-Opitz, Donald Gordon, Leander H. Shearer, Wm. K. Terriberry; Surgery: Hugh Auchincloss, William Darrach, Rolfe Kingsley, Adrian V. S. Lambert, F. T. Van Buren, Jr. ; Therapeutics: Maximilian Schulman; University Physician: Wm. H. McCastline; Vaudcrbilt Clinic: F. Morris Class, Julius W. Weinstein; Zoology: H. B. Goodrich, John D. Haseman, H. J. Muller, Charles Packard, Colleges. — Barnard: Helene M. Boas, Ruth S. Finch, Louise H, Gregory; College of Pharmacy: Charles W. Ballard; Teachers College: Louis E. Bisch, Jean Broadhurst, Ada M. Field, Mary G. McCormick, Mrs. A. P. McGowan, Sadie B. Vanderbilt. Advanced students. — Graduate (Coli, of P. and S.): Robert Bersohn, Isabel S. Dougherty, Frank R. Eider, H. Gertrude Gates, Hattie L. Heft, Mildred A. Hoge, Jacob Hoffman, Karl J. HoUiday, Margaret F. Kelley, Alexander Lowy, Darwin O. Lyon, Edward Plaut, Michael Puorro, G. S. Rosenthal, Fred L. Thompson, W. H. Schliffer, Jr., Arthur P. Tanberg, Jennie A. Walker. — Grad- uate (Teach. Coli.): Lucy Gillett, Greta Gray, Madelain Hahn, Hildegarde Knee- land, Helen McClure, Marguerite McLean, Ethel Ronzone, Margaret Stanton, Mary B. Stark. — Medical: M. W. Astarita, Louis Berman, Ernst Boas, David C. Bull, Will H. Chapman, Frederick Eberson, Joseph Felsen, L. H. Ferguson, Wm. Finkelstein, Joseph Goldstone, Julius Gottesman, Martin Holzman, W. S. Horton, Walter F. Hume, H. T. Hyman, Jerome Kohn, Jacob Lattman, J. A. Lazarus, Arnold Messing, M. V. Miller, F. B. Orr, H. J. Pyle, A. V. Salomon, Isidore Steinman, Harry J. Seiff, Jacob Shulansky, T. F. X. Sullivan, Henry A. Sussman, W. W. Tracey, J. L B. Vail, E. R Van Derwerker, E. R. Ware, Joseph Yampolsky, F. D. Zeman. College of Dental and Oral Surgery of N. Y. — Louise C. Ball, H. H. Janeway. CoRNELL University Medical College. — Stanley R. Benedict, Robert A. Cooke, Nellis B. Foster, Leon Loewe, Jessie Moore Rahe, Charles R. Stockard, Geo. W. Vandegrift. EcLECTic Medical College. — David Alperin. FoRDHAM University Medical College. — Benjamin Horowitz. Harriman Research Laboratory. — Isidor Greenwald. Hospitals. — Babies': Morris Stark; Bellevue: Edward C. Brenner, Edward M. Colie, Jr., W. M. Kraus, R. W. Lobenstine; Beth Israel: Charles J. Brim, Max Kahn, Alfred A. Schwartz; City: Louis Pine; Flower: Henry L. Weil; Flushing: Eimer W. Baker; General Memorial: Clinton B. Knapp; Gertnan: H. G. Baumgard, Melvin G. Herzfeld, Frederick B. Humphries, Charles H. Sanford, Fred S. Weingarten ; Hudson Street: Robert T. Corry ; Jewish: Abraham Ravich, Lebanon: Samuel Gitlow, M. J. Gottlieb, William Wein- berger; Lutheran: Daniel R. Lucas; Mt. Sinai: George Baehr, Samuel Book- man, Leo Buerger, Burrill B. Crohn, Simon S. Friedman, David J. Kaliski, John L. Kantor, Leo Kessel, Nathan Rosenthal, Harry Wessler; A''. Y.: Helen B. Davis, James C. Greenway, Ralph G. Stillman; A^^. Y. Nursery and Child's: Oscar M. Schloss; Presbyterian: Herbert S. Carter, Russell L. Cecil, Calvin B. Coulter; Roosevelt: J. Buren Sidbury; St. Luke's: Norman E. Ditman, W. S. Schley. Hunter College. — Beatrix H. Gross. Long Island Medical College. — Matthew Steel. Museum of Natural History. — Louis Hussakof, Israel J. Kligler. N. Y. Aquarium. — Raymond C. Osburn. N. Y. Association for Improving the Condition of the Poor. — Donald B. Armstrong. N. Y. BoTANicAL Garden. — Fred J. Seaver. N. Y. City Department of Education. — Boys' High School: Frank T. Hughes ; Brooklyn Training School: C. A. Mathewson ; Cotnmercial High School: W. J. Donvan, B. C. Gruenberg, Edgar F. Van Buskirk; DeWitt Clinton High School: Frank M. Wheat; Eastern District High School: Gertrude S. Burling- ham; Girls' High School: Marguerite T, Lee; High School of Commerce: Harvey B. Clough, Fred W. Hartwell; Jamaica High School: Ella A. Holmes, vi Charles H. Vosburgh; Manual Training High School: Anna Everson; Morris High School: Charles A. Wirth; Newtown High School: Nellie P. Hewins; Wadleigh High School: Helen Gavin, Elsie A. Kupfer, Helen G. Russell, Helen S. Watt. N. Y. City Department of Health. — Charles F. Bolduan, Alfred F. Hess. N. Y. Eye and Ear Infirmary. — Harold M. Hays. Members resident in New York (con.) N. Y. Medical College for Women. — Ella H. Clark. N. Y. Milk Committee, — Philip Van Ingen. N. Y. PoLYCLiNic Medical School. — Jesse G. M. Bullowa, Mabel C. Little. N. Y. State Food Inspection Laboratory. — Louis J. Hirschleifer. Post Graduate Medical School. — Arthur F. Chace. Pratt Institute. — Grace MacLeod. RoCKEFELLER INSTITUTE. — Alfred E. Cohn, George W. Draper, Frederic M. Hanes, Michael Heidelberger, Gustave M. Meyer, Hardolf Wasteneys. Russell Sage Institute of Pathology. — Eugene F. DuBois. TuRCK Institute. — Katherine R. Coleman, Anton R. Rose. Vettin School. — Laura I. Mattoon. E. V. Delphey, 400 West 57th Street, Manhattan; Leopold L. Falke, 5316 Thirteenth Avenue, Brooklyn; Mabel P. FitzGerald, 416 East 65th Street, Man- hattan; Abraham Gross, c/o Arbuckle Sugar Co., Brooklyn; Leon M. Herbert, 29 Canal Street, Manhattan ; Julius Hyman, 62 East poth Street, Manhattan ; Alfred H. Kropff, 619 Kent Avenue, Brooklyn; Shojiro Kubushiro, 86 Lexington Avenue. Manhattan, Non-Resident Members Agnes Scott College (Decatur, Ga.). — Mary C. de Garmo. Albright College (Myerstown, Pa.). — George H. Whiteford. Allegheny General Hospital (Pittsburgh). — James P. McKelvy. Carnegie Institution (Cold Spring Harbor, L. L). — Ross A. Gortner. Clark University (Worcester, Mass.). — Sidney Liebovitz. Drake University Medical School (Des Moines, la.). — E, R. Posner. Forest School (Biltmore, N. C). — Homer D. House. Georgia Experiment Station (Experiment). — C. A. Wells. Indiana Agricultural Exp. Sta. (LaFayette). — Fred D. Fromme. Iowa University Hospital (Iowa City). — Louis Baumann. Isolation Hospital (San Francisco, Cal.). — L. D. Mead. Jefferson Medical College (Phila.). — P. B. Hawk, Edward A. Spitzka. Johns Hopkins University (Baltimore). — John Howland, Burton R Livingston, Edwards A. Park. Kansas State Agricultural College (Manhattan). — Ula M. Dow. MacDonald College (Quebec). — Kathryn Fisher. Mass. Agricultural College (Amherst). — H. D. Goodale. Mayo Clinic (Rochester, Minn.).— Edward C. Kendali. Milwaukee County Agric. School (Wauwatosa, Wis.). — Alice H. McKinney. New Hampshire College (Durham). — Helen B. Thompson. New Mexico Agricultural College (State College). — R. F. Hare. N. J. Agricultural Experiment Station (New Brunswick). — Carl A. Schwarze, Guy West Wilson. N. Dakota Agricultural College (Agricultural College).— H. L. White. North Hudson Hospital (Weehawken, N. J.). — A. E. Olpp. Ohio Agricultural Experiment Station (Wooster), — A. D. Selby. Princeton University. — E. Newton Harvey. Psychopathic Hospital (Boston). — Herman M. Adler. Rensselaer Polytechnic Institute (Troy, N. Y.). — Fred W. Schwartz. vii Rochester A and M Institute. — Elizabeth G. Van Home. Rockford College (Rockford, 111.)- — Anna M. Connelly. Secondary Schools. — Brock Port State Normal School (N. Y.) : Ida C. Wads- worth; Itidiaua State Normal School (Tcrre Haute): Roscoe R. Hyde; Inglc- side School (New Milford, Conn.) : Mary L. Chase; Kiiox School (Tarrytown, N. Y.) : Clara G. Miller; New Bedford Jiidustrial School (Mass.): Constance C. Hart; North Texas State Normal School (Benton): Blanche E. Shaflfer; Fassaic High School (N. J.) : Hazel Donham, Helene M. Pope; Rochester High School (N. Y.) : David F. Renshaw; State Normal School (Truro. N. S.) : Blanche E. Harris. Texas A and M College (College Station). — M. K. Thornton. Trinity College (Hartford, Conn.).— Edward C. Stone, R. M. Yergason. TuLANE University (Ncw Orleans, La.). — Allan C. Eustis. U. S. Department of Agriculture (Wash.). — Carl L. Aisberg, W. N. Berg, H. E. Buchbinder, George A. Geiger, William Salant, Clayton S. Smith. U. S.FooD and Drug Inspection Laboratory (Phila.). — Harold E. Woodward. U. S. Food- Research Laboratory (Phila.). — Ernest D. Clark, Joseph S. Hepburn. University of Alabama Medical School (Birmingham). — Richard A. Bliss. University of California (Berkeley). — William T. Home. University of Chicago. — Mathilde Koch. University of Georgia Medical School (Atlanta). — ^William D. Cutter. University of Illinois (Urbana). — George D. Beal, Isabel Bevier, A. D. Emmett; College of Medicine (Chicago). — Edgar G. Miller, Jr., Grover Tracy, William H. Welker. University of Indiana (Bloomington). — Clarence E. May. University of Kentucky (Louisville). — Mary E. Sweeny. University of Manitoba (Winnipeg, Can.). — A. T. Cameron. University of Michigan (Ann Arbor). — A. Franklin Shull. University of Minnesota (Minneapolis). — Josephine T. Berry, Louise McDanell. University of Missouri (Columbia). — Louis E. Wise. University of Montana (Missoula). — J. E. Kirkwood. University of Pennsylvania (Phila.). — A. N. Richards. University of Porto Rico (Rio Piedras). — L. A. Robinson. University of Tennessee (Memphis). — Edwin D. Watkins. University of Texas (Austin). — Mary E. Gearing, Anna E. Richardson. University of Toronto (Canada). — Olive G. Patterson. University of Utah (Salt Lake City). — H. A. Mattill. University of Washington (Seattle). — Elizabeth Rothermel. University of Wisconsin (Madison). — Max Morse, W. H. Peterson. Vassar College (Poughkeepsie, N. Y.). — Cora J. Beckwith. Wesleyan University (Middletown, Conn.). — David D. Whitney. West Pennsylvania Hospital (Pittsburgh). — J. Bronfenbrenner, Jacob Rosenbloom. Williams College (Williamstown, Mass.). — John S. Adriance. Women's Affiliated Colleges of Delaware (Newark). — Winifred J. Robinson. Yale University (New Haven, Conn.). — Lorande Loss Woodruff. Sidney Born, Lemp Brewing Co., St. Louis, Mo.; Emma A. Buehler, New- ark, N. J. ; Edward G. Griffin, Albany, N. Y. ; Marston L. Hamlin, Yonkers, N. Y. ; F. C. Hinkel, Utica, N. Y. ; Cavalier H. Joüet, Roselle, N. J. ; Emile F. Kropf, Standard Chemical Co., Pittsburgh, Pa. ; Adeline H. Rowland, Pittsburgh, Pa. ; H. J. Spencer, Factoryville, Pa. ; William A. Taltavall, Redlands, Cal. ; David C. Twichell, Saranac Lake, N. Y. ; Isabel Wheeler, Toledo, Ohio. • •• Vlll EDITORS OF THE BIOCHEMICAL BULLETIN The editorial committee with the coUaboration of the members and the SPECIAL CONTRIBUTORS: DR. JOHN AUER, Rockefeiler Institute for Medical Research, N. Y. PROF. WILDER D. BANCROFT, Cornell University, Ithaca DR. A. M. BANTA, Carnegie Sta. for Exp. Evolution, Cold Spring Harbor, L. I. DR. N. R. BLATHERWICK, Yale University, New Haven, Conn. MR. H. C. BIDDLE, Urhana, III. DR. WALTER L. GROLL, Elisabeth Steel Magee Hospital, Pittsburgh, Pa. DR. CHARLES A. DOREMUS, 55 W. 53d St., New York City DR. ARTHUR W. DOX, Iowa State College Agric. Experiment Station, Arnes PROF. JOSEPH ERLANGER, Washington Univ. Medical School, St. Louis DR. J. M. EVVARD, Iowa State College Agric. Exp. Station, Arnes. DR. EPHRAIM M. EWING, N. Y. Univ. and Bellevue Hosp. Med. College DR. LEWIS W. FETZER, U. S. Dep't of Agriculture, Washington, D. C. PROF. MARTIN H. FISCHER, University of Cincinnati DR. MARY LOUISE FOSTER, Smith College, Northampton, Mass. PROF. J. E. GREAVES, Utah Agricultural College, Logan DR. S. C. GUERNSEY, Iowa State College Agric. Exp. Station, Arnes. DR. FRANK L. HALEY, Univ. of Alabama Med. Seh., Birmingham. DR. V. J. HARDING, McGill University, Montreal, Canada DR. R. H. M. HARDISTY, McGill University, Montreal, Canada DR. J. A. HARRIS, Carnegie Sta. for Exp. Evolution, Cold Spring Harbor, L. I. DR. K. A. HASSELBALCH, Finsen Institute, Copenhagen, Denmark PROF. G. O. HIGLEY, Ohio Wesleyan University, Delaware DR. S. L. JODIDI, U. S. Dep't of Agriculture, Washington, D. C. DR. VERNON K. KRIEBLE, McGill University, Montreal, Canada PROF. FRANCIS E. LLOYD, McGill University, Montreal, Canada PROF. JOHN A. MANDEL, N. Y. Univ. and Bellevue Hospital Med. College PROF. WILLIAM MANSFIELD, College of Pharmacy, Columbia University PROF. ALBERT P. MATHEWS, University of Chicago PROF. SHINNOSUKE MATSUNAGA, University of Tokyo, Japan DR. S. J. MELTZER, Rockef eller Institute for Medical Research, N. Y. PROF. VICTOR C. MYERS, N. Y. Post-Graduate Med. School and Hospital DR. RAY E. NEIDIG, Iowa State College Agric. Experiment Station, Ames DR. THOMAS B. OSBORNE, Conn. Agric. Experiment Station, New Haven MR. EMIL OSTERBERG, Cornell University Medical College, New York City DR. AMOS W. PETERS, The Training School, Vineland, N. J. DR. I. K. PHELPS, U. S. Dep't of Agriculture, Washington, D. C. PROF. R. H. A. FLIMMER, University College, London DR. W. EUGENE RUTH, Iowa State College Agric. Experiment Station, Ames PROF. R. F. RUTTAN, McGill University, Montreal, Canada DR. JESSE A. SANDERS, University of Indiana, Bloomington DR. J. J. SKINNER, Bureau of Soils, U. S. Dep't of Agric, Wash., D. C. DR. E. E. SMITH, 50 East 4ist St., New York City DR. A. E. SPAAR, City Hospital, Trincomalee, Ceylon PROF. UMETARO SUZUKI, University of Tokyo, Japan DR. CLARENCE J. WEST, Rockefeiler Institute for Medical Research, N. Y. MISS ANNA W. WILLIAMS, State Agric. Coli., Manhattan, Kan. PROF. E. WINTERSTEIN, Polytechnic Institute, Zürich, Switzerland DR. JULES WOLFF, Pasteur Institute, Paris ix OFFICERS OF THE COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION 1913--1914 HONORARY OFFICERS Fast Presidents: i9io-'i2: Prof. A. N. Richards, University of Penn., Phila. 1912-13: Prof. P. B. Hawk, Jefferson Medical College, Phila, President: Dr. Carl L. Alsberg, U. S. Bureau of Chemistry, Washington, D. C. Vice Presidents: Dr. Hugh Auchincloss, Columbia University Dr. William B. Boyd, College of the City of New York Prof. Mary E. Gearing, University of Texas, Austin Dr. James C. Greenway, New York Hospital, New York City Prof. Mary E. Sweeny, University of Kentucky, Louisville ACTIVE OFFICERS President, Prof. S. R. Benedict, Cornell University Medical School Vice President, Dr. F. T. Van Beuren, Columbia University Sccretary, Dr. Alfred P. Lothrop, Columbia University Treasurer, Prof. William J. Gies, Columbia University Executive committee — Prof. S. R. Benedict, Prof. William J. Gies, Dr. F. G. Goodridge, Prof. Paul E. Howe, Dr. A. P. Loth- rop, Dr. H. O. Mosenthal, Dr. F. T. Van Beuren. Editorial Committee: See the back of the title page. SUMMARY OF CONTENTS: VOL. III, Nos. ^12 No. 9. October, 1913 PAGE A MoDiFiED Hempel Gas Pipette. Stanley R. Benedict i The Influence of Arsenic upon the Biological Transformation of NiTROGEN IN SoiLS. /. E. Grcaves 2 The Nature of Humus and Its Relation to Plant Life. 5". L. Jodidi. . . 17 Cleavage of Benzoylalanine and Acetylglycine by Mold Enzymes. Arthur W. Dox and W. Eugene Ruth. 23 A Color Reaction of Glycine when Boiled with Chloral Hydrate. Edwin D. IVatkins. 26 Studies on Water Drinking: 15. The Output of Fecal Bacteria as Influenced by the Drinking of Distilled Water at Meal Time. N. R. Blatherwick and P. B. Hawk. 28 A Note on the Determination of Ammonia in Urine. Stanley R. Benedict and Emil Osterberg. 41 Studies of Aeration Methods for the Determination of Ammonium Nitrogen : 3. The Ammonium Nitrogen in Beef. Jacob Shulansky and William J. Gies. 45 A Study of the Influence of Cold-storage Temperatures upon the Chemical Composition and Nutritive Value of Fish. Clayton S. Smith. 54 A Further Study of the Chemical Composition and Nutritive Value of Fish Subjected to Prolonged Periods of Cold Storage. William A. Perlzweig and William J. Gies. 69 The Influence of Chronic Undernutrition on Metabolism. Sergius Morgulis. 72 Nitrogen Metabolism During Chronic Underfeeding and Subsequent Realimentation. Sergius Morgulis 74 Proceedings of the Biological Section of the American Chemical Society, Rochester, N. Y., September 10-12, 1913 : I.Executive Proceedings. /. K. Phelps, Secretary 76 2. Chairman's Address. Carl L. Aisberg, Chairman 77 3- Scientific Proceedings (Abstracts). /. K. Phelps, Secretary 80 The Biochemical Society, England 96 The Agricultural Colleges and Experiment Stations in the United States (continued on page 275) . A. C 98 Biochemical Bibliography and Index. William A. Perhweig 103 Biochemical News, Notes and Comment: General 112 Columbia University Biochemical Association 129 Columbia Biochemical Department 131 Editorials : Sir Oliver Lodge on " Continuity " 133 The Mathews Plan for an American Biological Society 134 The Mathews Plan : A Summary of Published Opinions 142 " Hormones " 148 xi No. 10. January, 1914 PAGE Dinner to Henry Hurd Rusby : The Alumni Association of the College OF Pharmacy of the City of New York Honors Dean Rusby (with Portrait) . William Mansficld MQ View-points in the Study of Growth. Lafayette B. Mendel 156 The Physico-chemical Basis of Striated-muscle Contraction : 3. The Maximum Surface Tension in Striated Muscle. William N. Berg. 177 4. Sources of Surface Tension in Striated Muscle. William N. Berg... 187 Researches on the Physico-chemical Properties of Vegetable Saps: 2. Note on a Comparison of the Physico-chemical Constants of the Juice of Apples and Pears of Varying Size and Fertility. /. Arthur Harris and Ross Aiken Gortner. 196 Studies of Plant Growth in Heated Soil. Guy West Wilson 202 A RE\^Ew of Methods for the Isolation and Identification of the Organic Constituents of Soils. A. W. Thomas 210 A Review of Regent Investigations on the Mineral Nutrition of Fungi. Arthur W. Dox. 222 A Review of Willstätter's Researches on Chlorophyll. Clarence J. West. 229 Tables of the Relative Depression of the Freezing Point, 1860/A, to Facilitate the Calculation of Molecular Weights. /. Arthur Harris and Ross Aiken Gortner. 259 The Influence of Underfeeding and of Subsequent Abundant Feeding on the Basal Metabolism of the Dog. Sergius Morgulis 264 The Ninhydrin Reaction. Paul E. Howe 269 A Rapid Clinical Test for Hyperglycemia. S. Gitlow and B. Horowitc. 272 The Agricultural Colleges and Experiment Stations in the U. S. (continued f rom page 98) . A. C 275 Proceedings of the First Annual Meeting of the Federation of Ameri- can Societies for Experimental Biology, in Philadelphia, December 2&-31, 1913. Paul E. Howe 276 Proceedings of Societies Meeting in Conjunction with the Federation of American Societies for Experimental Biology; and of the So- ciety OF American Bacteriologists. Paul E. Howe 294 The Biochemical Society, England. R. H. A. Plimmer, Secretary 301 Scientific Procfjedings of the Columbia University Biochemical Asso- ciation. Alfred P. Lothrop, Secretary 302 Biochemical Bibliography and Index. William A. Perlzweig 315 Biochemical News, Notes and Comment: General 323 Columbia University Biochemical Association 329 Columbia Biochemical Department , 335 Editorials : Federation of American Societies for Experimental Biology Z37 The Mathews plan for an American Biological Society 344 " Stimulants " 344 xü Nos. II-I2. April and July, 1914 PAGE Professor Hugo Kronecker (with portrait). S. J. Meltzer 345 The Viscosity of Bile. R. Burton-Opits 35i Notes on the Toxicity of Dilute Solutions of Certain Phenolic Com- pounds, AS Indicated by their Effect on Amphibian Eggs and Em- bryos, Together with References on Modifications of Pigment De- velopment. Ross Aiken G ortner and Arthur M. Banta 357 The Digestibility of Maize Consumed by Swine. S. C. Guernsey and John M. Evvard. 369 Experience with the Abderhalden Serum Test for Pregnancy. Jacob Rosenbloom. 373 A Note on the Use of Purified Antigen of Besredka in the Serum Diagnosis of Tuberculosis. /. Bronfcnbrenner and J. Rockman 375 The Diagnostic Value of the Landau Test for Syphilis. /. Bronfcnbrenner and J. Rockman. 377 Further Studies on Besredka Tuberculin. /. Bronfenbrenner and J. Rockman. 381 Studies on So-called Protective Ferments : I. The Sensitization of Substratum for the Abderhalden Test. J. Bronfenbrenner, W. T. Mitchell, Jr., and M. J. Schlesinger. 386 Effect of Salicylic Aldehyde on Plants in Soil and Solution Cultures. /. /. Skinner. 390 On the Phosphorus Content of Starch. A. W. Thomas 403 A Standard for the Determination of Ammonia by Means of Nessler Solution. Anton R. Rose and Katherine R. Coleman 407 A Micro-urease Method for the Determination of Urea. Anton R. Rose and Katherine R. Coleman. 411 Fasting Studies: 14. The Elimination of Urinary Indican During Two Fasts of Over One Hundred Days Fach. Carl P. Sherwin and Philip B. Hawk 416 Studies in Water Drinking : 20. The Relationship of Water to Certain Life Processes and More Es- pecially to Nutrition. P. B. Hawk 420 Muscular Work and the Respiratory Quotient. Sergius Morgulis 43S Bleached Flour. Frank L. Haley 440 Meetings of the Biological Division of the American Chemical Society, Cincinnati, Ohio, April 8 and g, 1914. Isaac King Phelps, Secretary. 444 The Biochemical Society, England. R. H. A. Plimmer, Secretary 452 Scientific Proceedings of the Columbia University Biochemical Asso- ciation. Alfred P. Lothrop, Secretary 454 Doctorates in Biological Chemistry. P. H. D 472 Biochemical Bibliography and Index. W. A. Perlsweig 475 Biochemical News, Notes and Comment: General 489 Journals 501 Institutes 505 War Notes 507 Columbia University Biochemical Association 511 Columbia Biochemical Department 517 xui Editorials : Hugo Kronecker (with portrait) 523 Delay in the Issue of the Biochemical Bulletin 524 Causes of the Clotting of Blood 524 Lipins : a Matter of Terminology 525 Creatin Content in Muscle 527 On the Constitution of Matter 529 Remarks on Research 531 BooKs Received 537 Index: Volume III. (Includes names of authors, and impersonal and per- sonal subjects) 545 Title Page for Vol. III, with Summary of Contents, List of Illustra- TioNS, etc i-xvi ZIV Alphabetic list of authors named in the foregoing summary of Contents (See author index — page 545 — for additional names of authors of abstracts, quotations, comment, etc.) A. C, 98, 275 Alsberg, CL, 77 Banta, am, 357 Benedict, SR, i, 41 Berg, WN, 177, 187 Blatherwick, NR, 28 Bronfenbrenner, J, 375, 377, 381, 386 Burton-Opitz, R, 351 CoLEMAN, KR, 407, 411 Dox, AW, 23, 222 EvvARD, JM, 369 GiES, WJ, 45, 69 GiTLOw, S, 272 GORTNER, RA, 196, 259, 357 Greaves, je, 2 GUERNSEY, SC, 369 Haley, FL, 440 Harris, JA, 196, 259 Hawk, PB, 28, 416, 420 HOROWITZ, B, zjz HowE, PE, 269, 276, 294 JoDiDi, SL, 17 LoTHROP, AP, 302, 454 Mansfield, W, 149 Meltzer, SJ, 345 Mendel, LB, 156 Mitchell, WT, 386 Morgulis, S, T2, 74, 264, 435 Osterberg, E, 41 Perlzweig, WA, 69, 103, 315, 475 P. H. D, 472 Phelps, IK, 76, 80, 444 Plimmer, RHA, 301, 452 RocKMAN, J, 375, 377, 381 Rose, AR, 407, 411 rosenbloom, j, 373 Ruth, WE, 23 Schlesinger, MJ, 386 Sherwin, CP, 416 Shulansky, J, 45 Skinner, JJ, 390 Smith, CS, 54 Thomas, AW, 210, 403 Watkins, ED, 26 West, CJ, 229 Wilson, GW, 202 XV VAGE LIST OF ILLUSTRATIONS Three portraits and seven plates No 9. OCTOBER, 1913 Plate I. A modified Hempel gas pipette (Benedict) 1 No. 10. JANUARY, 1914 Portrait. Henry H. Rusby I49 Plate 2. Comparison of physico-chemical constants of the Juices of apples and pears of varying size and fertility (Harris and Gortner) 201 Plates 3-5. Plant growth in heated soil (Wilson) 204 Nos. 11-12. APRIL AND JULY, 1914 Portrait. Hugo Kronecker 345 Plates 6-7. Effect of salicylic aldehyde on plants in soil and Solution cul- tures (Skinner) 39° Portrait. Professors Hugo Kronecker and Leon Asher, and some of their pupils, in the Physiological Institute, Bern ; July, 1899 523 XVl Vol. III October, 1913 No. 9 Biochemical Bulletin Edited, for the Columbia University Biochemical Association, by tho EDITORIAL COMMITTEE: HERMAN M. ADLER, JOHN S. ADRIANCE, DAVID ALPERIN. CARL L. ALSBERG, D. B. ARMSTRONG, GEORGE BAEHR, CHAS. W. BALLARD, LOUIS BAUMANN, GEORGE D. BEAL, S. R. BENEDICT, WM. N. BERG, JO'PHINE T. BERRY, ISABEL BEVIER, LOUIS E. BISCH, A. RICHARD BLISS, CHAS. F. BOLDUAN, SAMUEL BOOKMAN, SIDNEY BORN, WM. B. BOYD. J. B'FEN BRENNER, JEAN BROADHURST, LEO BUERGER, GER'DE BURLINGHAM J. G. M. BULLOWA, R. BURTON-OPITZ, A. M. BUSWELL, R. F. CALVERT, A. T. CAMERON, HERBERT S. CARTER, ARTHUR F. CHACE, ELLA H. CLARK, ERNEST D. CLARK, BURRILL B. CROHN, LOUIS J. CURTMAN, WILLIAM D. CUTTER, C. A. DARLING, WILLIAM DARRACH, NORMAN E. DITMAN, EUGENE F. DuBOIS, JAMES G. DWYER, WALTER H. EDDY, GUSTAVE EGLOFF, A. D. EMMETT, ALLAN C. EUSTIS, BENJ. G. FEINBERG, RUTH S. FINCH, HARRY L. FISHER, KATHRYN FISHER, NELLIS B. FOSTER, FRED D. FROMME, C. STUART GAGER, MARY C. de GARMO, MARY E. GEARING, WILLIAM J. GIES, Treasurer, SAMUEL GITLOW, A. J. GOLDFARB, H. D. GOODALE, H. B. GOODRICH, F. G. GOODRIDGE, ROSS A. GORTNER, ISIDOR GREENWALD, JAMES C. GREENWAY, LOUISE H. GREGORY, MARSTON L. HAMLIN, FREDERIC M. HANES, R. F. HARE, TULA L. HARKEY, T. STUART HART, E. NEWTON HARVEY, P. B. HAWK, M. HEIDELBERGER, JOSEPH S. HEPBURN, ALFRED F. HESS, L. J. HIRSCHLEIFER, BENJ. HOROWITZ, HOMER D. HOUSE, PAUL E. HOWE, Secretary, LOUIS HUSSAKOF, ROSCOE R. HYDE, HENRY H. JANEWAY, MAX KAHN, JOHN L. KANTOR, EDW. C. KENDALL, J. E. KIRKWOOD, ISRAEL J. KLIGLER, ARTHUR KNUDSON, MATHILDE KOCH, MARGUERITE T. LEE, VICTOR E. LEVINE, CHARLES C. LIEB, B. E. LIVINGSTON, ALFRED P. LOTHROP, Chairman, DANIEL R. LUCAS, WM. H. McCASTLINE, MARY G. McCORMICK, LOUISE McDANELL, GRACE MacLEOD, C. A. MATHEWSON, H. A. MATTILL, CLARENCE E. MAY, GUSTAVE M. MEYER, E. G. MILLER, JR., SERGIUS MORGULIS, MAX MORSE, H. O. MOSENTHAL, HERM'N J. MULLER, ARCHIBALD E. OLPP, B. S. OPPENHEIMER, RAY'D C. OSBURN, R. OTTENBERG, OLIVE PATTERSON, W. A. PERLZWEIG, W. H. PETERSON, LOUIS PINE, E. R. POSNER. P. W. PUNNETT, JESSIE MOORE RAHE, A. N. RICHARDS, ANNA RICHARDSON, WIN'F'D J. ROBINSON, ANTON R. ROSE. JAC. ROSENBLOOM, WILLIAM SALANT, OSCAR M. SCHLOSS, H. von W. SCHULTE, FRED W. SCHWARTZ, C. A. SCHWARZE, EMILY C. SEAMAN, FRED J. SEAVER, A. D. SELBY, A. FRANKLIN SHULL, CLAYTON S. SMITH, EDWARD A. SPITZKA, MATTHEW STEEL. RALPH G. STILLMAN, CHAS. R. STOCKARD, EDWARD C. STONE, MARY E. SWEENY, ARTHUR W. THOMAS, F. T. VAN BUREN, ELIZ. G. VAN HÖRNE, PHILIP VAN INGEN, CHAS. H. VOSBURGH, HAR'PH WASTENEYS, EDWIN D. WATKINS, WM. WEINBERGER, F. S. WEINGARTEN, J. W. WEINSTEIN, CHARLES WEISMAN, WM. H. WELKER, C. A. WELLS, HARRY WESSLER, H. L. WHITE, DAVID D. WHITNEY, ETHEL WICKWIRE, HERBERT B. WILCOX, GUY W. WILSON, LOUIS E. WISE, WM. H. WOGLOM, L. L. WOODRUFF, H. E. WOODWARD, HANS ZINSSER. NEW YORK Entered as second-clasa matter in the Post Office at Lancaster, Pa. MEMBERS OF THE COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION Honorary Members PROF. R. H. CHITTENDEN, First Director of the Columbia University De- partment of Biological (Physiological) Chemistry and Director of the Sheffield Scientific School of Yale University PROF. HUGO KRONECKER, Director of the Physiological Institute, Uni- versity of Bern, Swiiserland PROF. SAMUEL VV. LAMBERT, Dean of the Columbia University School of Mediane DR. JACQUES LOEB, Member of the Rockefeller Institute for Medical Re- search and Hcad of the Department of Expcrimental Biology PROF. LAFAYETTE B. MENDEL, Professor of Physiological Chemistry, Sheffield Scientific School, Yale University PROF. ALEXANDER SMITH, Head of the Department of Chemistry, Co- lumbia University Corresponding Members PROF. LEON ASHER, University of Bern, Switzerland PROF. FILIPPO BOTTAZZI, University of Naples, Italy PROF. ROBERT B. GIBSON, University of the Philippines, P. I. PROF. VLADIMIR S. GULEVIC, University of Moscow, Russia PROF. W. D. HALLIBURTON, King's College, London PROF. S. G. HEDIN, University of Upsala, Sweden PROF. FREDERICO LANDOLPH, University of La Plata, Argentina PROF. A. B. MACALLUM, University of Toronto, Canada PROF. D. McCAY, Medical College, Calcutta, India PROF. C. A. PEKELHARING, University of Utrecht, Holland PROF. S. P. L. SÖRENSEN, Carlsberg Laboratory, Copenhagen, Denmark Members Resident in New York City Brooklyn Botanic Garden. — C. Stuart Gager, College of the City of New York. — Wm. B. Boyd, Louis J. Curtman, Benj. G. Feinberg, A. J. Goldfarb. Columbia University: Departments.— Anatomy: Alfred J. Brown, H. von W. Schulte; Bacteriology: James G. Dwj'er, Reuben Ottenberg, Hans Zinsser; Biological Chemistry: Walter H. Eddy, William J. Gies, F. G. Goodridge, Tula L. Harkey, Paul E. Howe, Arthur Knudson, Alfred P. Lothrop, Sergius Morgulis, H. O. Mosenthal, W. A. Perlzweig, Emily C. Seaman, Chris Seifert, Ethel Wick- wire; Botany: E. R. Altenburg, C. A. Darling; Cancer Research: W. H. Wog- lom; Chemistry: Sidney Born, A. M. Buswell, R. P. Calvert, Gustave Egloflf, H. L. Fisher, P. W. Punnett, A. W. Thomas; Clinical Pathology: Edward Cussler, Peter Irving, Arthur W. Swann; Diseases of Children: Chas. Hendee Smith, Herbert B. Wilcox; Gynecology: Wilbur Ward; Mediane: T. Stuart Hart, I. 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J. ; Emile F. Kropf, Standard Chemical Co., Pittsburgh, Pa. ; Adeline H. Rowland, Pittsburgh, Pa. ; H.J.Spencer, Factoryville, Pa. ; William A. Talta- vall, Redlands, Cal. ; David C. Twichell, Saranac Lake, N. Y. ; Isabel Wheeler, Toledo, Ohio., ANNOUNCEMENT. Professional Assistance Offered to Biological Chemists The members of the Columbia University Biochemical Association will coöperate confidentially with any one who desires the Services of biological chemists or who seeks a position in biological chemistry. Address inquiries to William J. des, 457 West sgth St., New York. EDITORS OF THE BIOCHEMICAL BULLETIN The editorial committee with the collaboration o£ the members and the SPECIAL CONTRIBUTORS: DR. JOHN AUER, Rockefeiler Institute for Medical Research PROF. WILDER D. BANCROFT, Coniell Uitiversity, Ithaca MR. N. R. BLATHERWICK, Yale Univcrsify, Nezv Havcn, Conn. MR. H. C. BIDDLE, Urbana, III. DR. WALTER L. GROLL, Elisabeth Steel Magee Hospital, Pittsburgh, Pa. DR. CHARLES A. DOREMUS, 55 W. 53d St., New York City DR. ARTHUR W. DOX, Iowa State College Agric. Experiment Station. Arnes PROF. JOSEPH ERLANGER, Washington Univ. Medical School, St. Louis DR. EPHRAIM M. EWING, A^. Y. Univ. and Bellevue Hosp. Med. College DR. LEWIS W. FETZER, U. S. Dep't of Agriculture, Washington, D. C. PROF. MARTIN H. FISCHER, University of Cincinnati DR. MARY LOUISE FOSTER, Smith College, Northampton, Mass. PROF. J. E. GREAVES, Utah Agricultural College, Logan DR. V. J. HARDING, McGill University, Montreal, Canada DR. R. H. AI. HARDISTY, McGill University, Montreal, Canada DR. J. A. HARRIS, Carnegie Sta. for Exp. Evolution, Cold Spring Harbor, L. I. DR. K. A. HASSELBALCH, Einsen Institute, Copenhagen, Denmark PROF. G. O. HIGLEY, Ohio Wesleyan University, Delaware DR. S. L. JODIDI, U. S. Dcp't of Agncultnre, Washington, D. C. DR. VERNON K. KRIEBLE, McGill University, Montreal, Canada PROF. FRANCIS E. LLOYD, McGill University, Montreal, Canada PROF. JOHN A. MANDEL, N. Y. Univ. and Bellevue Hospital Med. College PROF. ALBERT P. MATHEWS, University of Chicago PROF. SHINNOSUKE MATSUNAGA, University of Tokyo, Japan PROF. VICTOR C. MYERS, N. Y. Post-Graduate Med. School and Hospital DR. RAY E. NEIDIG, Iowa State College Agric. Experiment Station, Arnes DR. THOMAS B. OSBORNE, Conn. Agric. Experiment Station, New Haven MR. EMIL OSTERBERG, Corncll University Medical College, New York City DR. AMOS W. PETERS, The Training School, Vineland, N. J. DR. I. K. PHELPS, U. S. Dep't of Agriculture, Washington. D. C. DR. W. EUGENE RUTH, Iowa State College Agric. Experiment Station, Arnes PROF. R. F. RUTTAN, McGill University, Montreal, Canada DR. JESSE A. SANDERS, University of Indiana, Bloomington DR. E. E. SMITH, 50 East 4ist St., New York City DR. A. E. SPAAR, City Hospital, Trincomalee, Ceylon PROF. UMETARO SUZUKI, University of Tokyo, Japan MISS ANNA W. WILLIAMS, University of Illinois, Urbana, III. PROF. E. WINTERSTEIN, Polytechnic Institute, Zürich, Switzerland DR. JULES WOLFF, Pasteur Institute, Paris BiocHEMiCAL Bulletin (Vol. ilh Plate 1 BENEDICT: A MODIFIED HEMPEL GAS PIPETTE. BiocHEMiCAL Bulletin Volume III OCTOBER, 191 3 No. 9 A MODIFIED HEMPEL GAS PIPETTE STANLEY R. BENEDICT (Department of Chemistry,Cornell University Medical College, New York City) (WITH PLATE l) In the use of the Hempel pipette the shaking necessary to obtain complete absorption of a gas is undesirable from two Standpoints. It is time-consuming and, unless the connections are exactly right, air is apt to enter the apparatus during the protracted period of vigorous shaking which is often necessary. Van Slyke, in his latest apparatus for amino nitrogen determina- tion, employs a small motor for shaking the pipette, and thus obvi- ates the awkward procedure of shaking by hand. The modified form of apparatus here described was designed to eHminate entirely any necessity for shaking in the Hempel appa- ratus to secure complete absorption. The figure is almost self-explan- atory (Plate i). At p is a three-way stop-cock, through which the gas is allowed to enter b through the tube h. The gas thus bubbles through the Solution contained in b, collects at the top, and is drawn out through the tube d_, after turning p. The intimate mixture of gas and liquid effected by bubbling the gas through the Solution is sufficient to ensure a complete reaction if the process is repeated once er twice. We have used the apparatus in this laboratory in a large number of determinations, with highly satisfactory results. THE INFLUENCE OF ARSENIC UPON THE BIOLOG- ICAL TRANSFORMATION OF NITROGEN IN SOILS J. E. GREAVES (Utah Experiment Station, Logan) Arsenic occurs in many virgin soils and is repeatedly added to others in insecticides, in commercial fertilizers, or in the flue dust from various smelters. Some investigators hold that arsenic may accumulate in soil in quantities sufficient to injure Vegetation grown upon such soil, and that the plants may in turn poison animals which feed upon them. The work described in this paper was undertaken to see if the lower plants, the ammonifiers and nitrifiers, of the soil are injured by arsenic, for they are readily experimented with and yield results in much shorter time than higher plants. Such results may also indicate the way in which a study of the higher plants should be pursued, for it is probable that substances which influence the soil organisms may also influence higher plants, differing only in degree ; but results of this kind must be accepted only as indicative, not as absolute proof, of what may occur with higher plants, for as pointed out by Guthrie and Helms, ^ all plants are not equally :sensitive to poisons. I. INFLUENCE OF ARSENIC UPON AMMONIFICATION Method of experimentation. The work was carried on with a typical bench soil, a sandy loam, fairly high in calcium and iron content, and supplied with an abundance of all the essential ele- ments of plant food with the exception of nitrogen which was low, a characteristic of arid soils. The soil was air dried, sieved and stored in a large box, so that all determinations could be made on the same soil. Determination of the ammonifying powers was made by Lip- 1 Guthrie and Helms : Agr. Gas. New South Wales, 1903, xiv, p. 114. 2 I9I3] /. E. Greaves man's^ method, with slight modifications. Beakers covered with Petri dishes were sterilized and into these were weighed loo-gm. portions of the air dried soil and i gm. o£ dry blood. Sodium arsenate was added from a Standard Solution with the proper proportion of sterile water. The dry insoluble Compounds were thoroughly mixed with the soil and then the water content of the soil made up to i8 percent with distilled sterile water. The samples were incubated at 28° C. to 30° C. for four days and the ammonia determined by transferring to Kjeldahl flasks with 250 cc. of distilled water, adding 2 grams of magnesium oxide and distilling into n/io sulfuric acid sol. The determinations were made in duplicate and compared with sterile blanks, so that each reported result is the average of two er more closely agreeing determinations. The results for soluble arsenic (sodium arsenate) are given in Table i. TABLE I Data pertaining to the ammonia produced in 100 grams of soil containing dif- ferent amounts of arsenic in the form of sodium arsenate Arsenic; parts Milligrams of 1 Arsenic; parts Milligrams of Arsenic ; parts Milligrams of per raillion NH3 formed 1 per million NH3 formed per million NH3 formed None 48.02 I 48.24 10 4506 55 32.12 2 52.72 15 45-06 60 32.78 3 58.12 20 36.54 65 34-82 4 53-36 25 38.90 70 34-00 5 52.68 30 38.22 75 34-40 6 51.02 35 36.00 80 32.12 7 51.66 40 32.78 85 32.68 8 46.76 45 34-80 90 33-60 9 46.76 50 32.60 From the above results it may be seen that, in dilute Solution, sodium arsenate stimulated the activity of the ammonifying organ- isms of the soil, the greatest influence having been exerted at a con- centration of 3 parts per million. The eff ect was quite marked up to a concentration of 7 parts per million. Part of this stimulating action, however, may have been due to the anion and not to the cation, as Lipman in his work has noted a stimulating effect with some sodium Compounds. No retarding influence was noted untilthe concentration of the sodium arsenate reached 20 parts per million, 2 Lipman : Centralbl. f. BakterioL, 1912, xxxii, p. 8. 4 Influence of Ar senk in Solls [Oct. after which there was a marked retarding influence on the ammoni- fying powers of the soll. From this point the quantity of ammonia formed was nearly constant; only a shghtly greater retarding effect having been noted when the concentration of the arsenic was 90 parts per milHon than when it was 20. This is probably due to the fact that the soil was comparatively rieh in calcium and iron, and that the sodium arsenate was changed into the comparatively insol- uble calcium and iron arsenates. At the concentration of 20 parts per million the water in the soil was saturated with these slightly soluble Compounds, so that even though there was an increase in the quantities added, the active masses remained about constant. These results, therefore, cannot be taken to indicate what may happen in a soil devoid of calcium and iron, and in which the arsenic may re- main in a soluble form. They show that large quantities of soluble arsenic, even up to 90 parts per million, may be added to a soil rieh in calcium and iron without stopping ammonification. The influence of insoluble arsenic Compounds. The Com- pounds selected for the tests are in use as sprays, and hence find their way into the soil in greater or less quantities. They were lead arsenate, Paris green, zinc arsenite and arsenic trisulfide. The tri- sulfide, while seldom used, is of special interest in this work in that it permits the application of arsenic free from the metallic elements which accompany each of the others, and therefore gives more nearly the influence of the arsenic than do the other Compounds. In each case the quantity of the Compound taken was such as to give equivalent amounts of arsenic. The results reported are in milli- grams of ammonia formed per 100 grams of soil. These results are reported in Table 2. On examining the data in Table 2 we find that arsenate stim- ulated ammonification in the lowest concentrations and that the tox- icity gradually increased as the amount of lead arsenate in the soil was increased. For no concentration tested was there any marked retarding action over the previous concentration, but a gradual de- cline in the ammonifying efiiciency. The ammonia was not re- duced to one-half of its original amount until the soil contained a concentration of 760 parts of arsenic per million. Even at a con- centration of 1,120 parts per million of arsenic, there was produced I9I3] /. E. Greaves TABLE 2 Data pertaining to the ammonia produced in wo grams of soll containing dif- ferent amounts, and different forms, of arsenic Lead arsenate ; Paris green ; Zinc arsenite ; Arsenic trisulfide ; Arsenic added ; milligrams NH3 milligrams NH3 milligrams NH milligrams NH3 parts per million formed formed formed formed None 40.46 39-28 34-25 39-44 20 41.82 32.64 35-02 43-46 40 38.42 28.22 33-32 46.92 80 38.41 26.52 28.22 42.16 120 35-02 26.86 24.82 42.16 160 35-36 20.74 22.44 40.80 200 35-70 21.42 21.12 39-78 240 36.72 19.72 18.02 40.13 280 33-32 19.38 17-34 41.08 320 33-66 19.38 17-34 40.44 360 28.22 15.98 17.00 40.80 400 27.28 15-30 17.80 40.46 440 27.20 13-94 17-34 40.79 480 28.80 10.20 17.00 39-79 520 25.16 10.54 17-34 40.11 560 24-34 6.12 17.68 38.77 600 24-34 4.42 18.02 40.12 640 23-46 5-10 18.02 39-77 680 22. 78 4.42 17.68 39-44 ♦ 720 21.76 5-10 17-85 38.41 760 20.40 4-42 17.68 38.7s 800 20.00 4.42 17-51 39.10 840 20.40 4.42 17-51 38.41 880 20.40 4-42 17.68 38.72 920 20.40 4.76 17.68 38.08 960 19.09 4-42 17-34 37-74 1,000 19.04 4.76 15.64 37-40 1,040 18.36 3-16 15.64 36.71 1,080 19.02 2.72 15-81 36.70 1,120 15.20 0.68 15-30 36.38 over one-third of the amount of ammonia formed in the total absence of lead arsenate, showing that even this large quantity was not sufficient to kill or even to entirely stop the activity of the soll organisms. Paris green, on the other hand, acts as a strong poison to the ammonifying organisms. Before the concentration of arsenic in this form in the soil reached a concentration of 240 parts per mil- lion, the ammonifying efficiency of the soil was reduced one-half. Even 20 parts per million of arsenic retarded very materially the ammonifying powers of the soil. At the highest concentration tested (1,120 parts per million) practically no ammonia was formed. Arsenic trisulfide in the lowest concentration had a marked 6 Influence of Arsenic in Solls [Oct. stimulating effect on ammonification and no toxic influence was noted until the highest proportions were present. Even 1,120 parts of arsenic per million exerted little toxic influence. The zinc arsenite and lead arsenate are very similar in their action, for they both apparently stimulate the ammonifying organ- isms of the soil when used in small quantities ; and while in greater concentration they exert a certain toxic influence upon the soil or- ganisms, this influence increases slowly with the added quantity of the Salt. The power of the soil to produce ammonia, after 800 parts of arsenic in the form of lead arsenate had been added, was 49.43 percent of the power of the original soil, while with the same arsenic concentration in the form of zinc arsenite, it was reduced to 51. II percent. At the highest concentration tested, 1,120 parts per million, the lead arsenate reduced the ammonifying efficiency to 37-57 percent of the original soil while the zinc arsenite reduced it to 44.68 percent of the original soil. Furthermore, it may be Seen that neither of these salts retarded very materially the ammon- ifying powers of the soil when present in a quantity equal to that known to occur in soils.^^ This, however, is not the case when the arsenic is applied in the form of Paris green. This substance, even in the lowest concentration, retards very materially the ammon- ifying powers of the soil. II. INFLUENCE OF ARSENIC UPON NITRIFICATION Method of experimentation. Soil similar to that used in the ammonification tests was employed. The method was that of Lip- man.^** Beakers covered with Petri dishes were sterilized and into these were weighed loo-gm. portions of the air-dried soil and 2 gm. of dry blood. Sodium arsenate was added from a Standard Solu- tion with the proper proportion of sterile water, and the mixture thoroughly stirred with a sterile spatula. The so-called insoluble arsenates were added in the form of the dry powders and then thor- oughly mixed with a sterile spatula. Sufficient sterile distilled water was added to make the moisture content of the soil 18 percent. These portions were weighed and the moisture content made up weekly to 18 percent. 2aGreaves: Biochemical Bulletin, 1913, ii, p. 519. 21) Lipman : Centralhl. f. BakterioL, 1912, xxxii, p. 8. I9I3] /. E. Greaves The mixtures were incubated at 28° C. to 30° C. for three weeks and the nitrates determined by transferring the soil, by means of 250 c.c. of distilled water, to a mortar containing 2 gm. of quick lime. The soil was triturated for 2 minutes and then transferred to a closed bottle and allowed to stand for 12 hours. At the end of this time an aHquot portion, 25 c.c, was measured into a 100 c.c. beaker and evaporated to dryness. The residue was treated with 2 c.c. of phenol-disulphonic acid equally distributed over all the res- idue and then allowed to stand for 10 minutes. The resulting Solu- tion was diluted with water and the excess of acid neutralized with dil. ammonium hydroxid sol. The ensuing color was compared with that produced by a Standard Solution of potassium nitrate treated in the same manner. The determinations were all made in duplicate and compared with sterile blanks, so that each reported result is the average of two or more closely agreeing determina- tions. The results obtained for soluble arsenic (sodium arsenate) are given in Table 3. TABLE 3 Data pertaining to nitric nitrogen produced in 100 grams of soil containing dif- ferent amounts of arsenic in the form of sodium arsenate Arsenic; parts per million Milligrams of nitric nitrogen formed Arsenic; parts per million Milligrams of nitric nitrogen formed Arsenic ; parts per million Milligrams of nitric nitrogen formed None 9-7 10 9-5 55 9.8 I 9.6 IS 9.2 60 9.6 2 9.6 20 9.8 65 9.7 3 9.4 25 10.4 70 9-5 4 9.0 30 10.3 75 II. I 5 9.2 35 9.0 80 14-5 6 9.8 40 9.0 85 15.6 7 95 45 9.0 90 9.4 8 10.2 50 9.6 100 6.0 9 9.8 A striking general similarity is seen to exist between the results here presented and those found for the ammonification series. No toxicity was noted until 100 parts per million were present. It is likely that the main part of the soluble arsenic was transformed into the comparatively insoluble iron and calcium arsenates, hence the great similarity throughout the entire series. The Stimulation caused when the concentration was from 75 to 85 parts per million 8 Infliience of Arsenic in Solls [Oct was probably diie partly to the sodium ion. As Lipman^ has noted, there is a marked Stimulation of nitrification in soils when small quantities of either sodium sulphate or sodium chloride are applied to them. TABLE 4 Data pertaining to nitric nitrogen produced in wo grams of soll containing dif- ferent amounts, and different forms, of arsenic Lead arsenate ; Paris green ; Zinc arsenite; Arsenic trisulfide ; Arsenic added ; milligrams of nitric milligrams of nitric milligrams of nitric milligrams of nitric parts per million nitrogen formed nitrogen formed nitrogen formed nitrogen formed None 10.5 10.4 10.2 9-5 20 17 9 10.8 lO.S 10.2 40 18 7 9.4 10.8 "•3 80 14 5 9.6 10. 13-2 I20 13 S lO.O 9.8 II-3 i6o 13 5 9-7 10.3 11.9 200 12 5 9.9 9-3 10.2 240 II 5 10.4 10.3 10.2 280 9 5 13-4 9.7 11.6 320 10 2 15-4 9.4 9-7 360 10 3 14.4 9-9 9.2 400 9 8 13.4 10.8 9.6 440 9 7 14.0 9.0 10. 480 9 6 14.1 14.0 8.8 520 10 5 13-4 14.4 8.3 600 10 3 13-4 15.0 6.3 640 10 12.4 16.8 5-4 680 9 8 10.4 12. 1 6.5 720 9 5 11. 10. S-o 760 9 4 9.9 9.0 4.0 800 9 3 9-7 8.6 4.0 840 8 5 9.4 7.4 3-9 880 8 4 9.4 6.8 2.2 920 8 2 9.4 6.0 2.2 960 8 8.4 6.8 2.0 1,000 7 9 7-4 7.0 I.O 1,040 7 8 7.4 6.0 I.O 1,080 7 5 5-4 S-2 1.0 1,120 7 I 3-4 5-3 0.8 The influence of insoluble arsenic Compounds. The Com- pounds used in the ammonification tests were employed for this series : lead arsenate, Paris green, zinc arsenite and arsenic trisulfide. In each case the quantity of the Compound taken was such as to give equivalent amounts of arsenic. The results, reported as milligrams of nitric nitrogen per 100 grams of soil, are given in Table 4. These results bring out some very interesting facts, which are 3 Lipman : Centralbl. f. Bakteriol., 1912, xxxii, p. 8. 1913] J- E. Greaves 9 of considerable theoretical and practical importance. We note a marked stimulating effect with each of the substances used, which varied greatly with the different Compounds. For instance, there was great Stimulation with lead arscnate in the lowest concen- tration tested. At this concentration the quantity of resultant nitric nitrogen was nearly twice that produced in the untreated soil. As the concentration of the arsenic increased above240 parts per million there was a sHght decrease in the production of nitric nitrogen, but a concentration of even 1,120 parts of arsenic in the form of lead arsenate caused the production of nearly as much nitric nitrogen as was formed in the untreated soil. It is rather hard to decide, f rom these results, whether the observed Stimulation was due to the chem- ical applied or to a change in the physical condition of the soil. Probably both are concerned but only a small portion can be attrib- uted to a purely physical change in the soil. Examining the results obtained where the Paris green was used, we find a stimulating effect with the lower concentrations of arsenic, which reached its maximum when 320 parts per million of arsenic were present. Above this concentration it decreased and at a con- centration of 760 parts per million of arsenic the effect commenced to be toxic. The toxicity increased with the concentration until, at the highest concentrations, the production of nitric nitrogen was reduced to 3.4 mg. of nitric nitrogen per 100 gm. of soil. Zinc arsenite exerted no apparent influence on the formation of nitric nitrogen in the soil until the concentration was 480 parts per million; after which a notable stimulating influence was observed. This Stimulation continued until the concentration was 680 parts per million. Above this concentration there was a marked retarding influence upon nitrification. Arsenic tristdfide stimulated the formation of nitric nitrogen slightly until the concentration of the arsenic trisulfide exceeded 280 parts per million ; after which there was a marked falling off in the production of nitric nitrogen and at the highest concentration tested (1,120 parts per million), the formation of nitric nitrogen practically ceased. Paris green, zinc arsenite and arsenic trisulfide after their first slight stimulating influence, which varied in intensity with the dif- 10 Influence of Arsenic in Soils [Oct. ferent Compounds, exerted very marked toxic influences. This was greatest for arsenic trisulfide and least for zinc arsenite. It is im- probable that sufficiently large quantities of any of these Compounds (lead arsenate, zinc arsenite, arsenic trisulfide, Paris green), would be added to a soll under natural Systems of agriculture to very mate- rially retard nitrification. On the other band, these results Iudicata that a marked stimulating influence may be exerted. III. GENERAL CONSIDERATIONS Throughout this work there has been noted a wide difference in the quantitative action of arsenic Compounds, but a striking simi- larity in qualitative influence. Some exerted little influence in the lowest concentrations on either ammonification or nitrification, but in the higher concentrations exerted a very marked toxic influence, e. g., Paris green. All, at some concentration, exerted either a slight or very great stimulating influence. This fact suggests that arsenic in small quantities tends to stimulate the bacterial activity of the soil as measured by ammonification and nitrification. On the other band, the absence of any great toxic influence where the sol- uble arsenic was applied and the great variations in results noted with other Compounds, raise the question whether the toxic influence is exerted by the anion or cation. In Order to gain some light on this subject, determinations were made of the water-soluble arsenic in soil to which the various forms of arsenic were added. These determinations were made as fol- lows : Quantities of lead arsenate, Paris green, zinc arsenite, and arsenic trisulfide were added to loo-gm. portions of soil in quanti- ties sufiicient to give 1,120 parts of arsenic per million of soil. The soil and arsenic, together with 2 gm. of dry blood, were placed in sterile tumblers; the water content was made up to 18 percent and then incubated at 28° C. for three weeks. At the end of this time the soil was transferred by means of 1,000 c.c. of carbon dioxide-free, distilled water to large acid bottles. The mixture was left in these bottles with occasional shaking for eight days, then filtered, and the arsenic determined^ in an aliquot part. In another set the vari- ous forms of arsenic were mixed with lOO-gm. portions of soil * Greaves : Jour. Amcr. Chem. Soc, 1913, xxxv, p. 150. I9I3] /. E. Greaves II and 2 gm. of dry blood, and the water-soluble arsenic determined as above, without incubating. The average of the two sets of results should give a very close approximation of the quantity of water-soluble arsenic existing in the soil during the activity of the organisms. The results are given in Table 5, as milligrams of water-soluble arsenic occurring in 100 gm. of the soil (before and after three weeks' incubation), to which 1,120 parts per million of the various forms of arsenic were added. Each is the average of two or more closely agreeing determinations. TABLE 5 Data pertaining to water-soluble arsenic in wo grams of soil to which 1120 parts per million of arsenic were added Treatment Incubated three weeks. Water-soluble arsenic determined (mg.) Water-soluble arsenic determined directly (mg.) Average (mg.) Lead arsenate Paris green Zinc arse- nite Arsenic trisulfide 14-3 20.2 17-3 80.0 82.0 81.0 36.9 31-7 34-3 50.0 5.6 27.3 These results show that there are great differences in the quan- tities of water-soluble arsenic in a soil to which various forms of arsenic have been added, and that even soil rieh in iron and calcium to which arsenic has been added in large quantities may have a high water-soluble arsenic content. This content is highest when the arsenic is added in the form of Paris green. Paris green ap- parently becomes less soluble, while arsenic trisulfide becomes much more soluble, as nitrification takes place. Comparing these results with the ammonia and nitrates formed under the several conditions, we note a general relationship between the toxicity of the substance added and the quantity of water-soluble arsenic present. The great- est toxic effect is noted with Paris green and this gives the greatest amount of water-soluble arsenic. Part of this toxicity may be due to the copper ion, as it is well known that this substance acts as a strong poison to many of the lower plants. Miss Benchley^ found it to be toxic to higher plants sBenchley: Annais of Botany, 1910, xxiv, p. 571. 12 Influcnce of Arsenic in Solls [Oct. also, when present to the extent of only i part per ten million of water. Although Russell" states that it is not as toxic in soll as in water, he and Darkshire^ found it to be toxic in soils; and they failed to get a stimulating influence witli it. Montemartini^ has noted a Stimulation with copper sulphate when used in dilute Solu- tions. This, however, may have been due to the anion and not to the cation, as sulphates do stimulate plant growth by their action on insoluble constituents of the soil.^ Clark and Gage^° have found that very dilute Solutions of copper have an invigorating influence upon bacterial activity. In order that any Stimulation may be noted, the copper must be present in very small quantities. Jackson^ ^ found that i part of copper sulphate in 50,000 parts of water killed B. coli and B. typhosus. Kellerman and Beckwith^^ found that the common saprophytic bacteria are more resistant to copper than is B. coli; their results also show stimulating influences at some con- centrations. So it is likely that both the copper and arsenic stim- ulated in these experiments. With all the other Compounds tested there was either a marked or very slight Stimulation. This is in keeping with the findings of other investigators. Johnson^ ^ found that arsenic, in dilute Solutions, stimulated seeds, while Bouilhac^^ found that it stimulated algae. Stimulation of nitrification may be due either to the arsenic inhibit- ing or killing injurious species, or to a direct Stimulation. It seems remarkable that bacteria should function in the presence of the large quantities of water-soluble arsenic which were present in some of these tests. One would be prone to ascribe the nitrifica- tion to something other than a biological influence, were it not for the fact that Gosio^^ grew moulds in organic matter containing ß Russell : Soil Conditions and Plant Growth ; New York and London, 1912, p. 47. '' Russell and Darkshire : Jour. Agr. Sei., 1905, i, p. 261. 8 Montemartini : Bull. Agr. Bur. Intel, and Plant Diseases, 191 1, ii, p. 2467. 8 Greaves : Jour. Biological Chem., 1910, vii, p. 298. ^•^ Clark and Gage: Jour. Inject. Diseases, 1906, ii, p. 175. 11 Jackson : Jour. Anter. Chem. Soc, 1905, xxvii, p. 675. 12 Kellerman and Beckwith : U. S. Dep. Agr., Bur. Plant Ind., Bull. 100, p. 57. 13 Johnson: Exp. Sta. Record, i896-'97, viii, p. 232. i*Bouilhac: Ibid., iSg^-'oo, xi, p. 1916. i^Gosio: Lafar's Technical Mycology; Trans, by Salter, 1911, ii, p. 37. 1913] J- E,. Greaves 13 arsenic. In fact one, Penicilliiim hrevicaiile, was so adapted to grow in it and evolve diethylarsine that he proposed this as a means of detecting arsenic. Furthermore, one could easily pick out the mixtures containing the greater quantity of arsenic in this work, from the large quantities of mould upon their surfaces. The very great stimulating effect of the zinc arsenite in the nitri- fication series was probably due as much to the stimulating influ- ence of the zinc as to that of the arsenic. Lathan^® found that small quantities of zinc stimulated algae. The same results have been obtained by Silberberg^^ working with higher plants. Ehren- berg's^^ work is of special interest in this connection, as he found that zinc stimulated plant growth in soils ; but when the soil was ster- ilized, the zinc became toxic. This would indicate that the Stimula- tion which has been noted by many investigators is probably only an indirect infiuence; and when the soil organisms were killed by heat, the toxic influence on the plant became perceptible. Lead arsenate stimulates very greatly the nitrification in soil. Part of this Stimulation is most likely due to the arsenic while some is due to the lead. Stoklasa^*^ has shown that lead, when present in small quantities, stimulates the growth of higher plants. It is pos- sible that the amorphous lead arsenate improves the texture of the soil and in so doing increases nitrification in it. Little if any effect could be attributed to the purely catalytic influence of lead, as Russell and Smith^*^ found this to be very small. Again there is the possi- bility that some of the Compounds inhibit the activity of injurious species. But these possibilities, and the question as to how much the results would vary in other soils, are problems which can be solved only by further work. Stoklasa^^ ascribed the observed Stimulation of growing sugar beets, when arsenic and lead were applied to the soil, to the catalytic action of these Clements on the chloroph>dl apparatus of the plants. These results indicate that it was due to their influence on the biological transformation of the nitrogen in the soil. 16 Lathan : Bul. Torrey Bot. Club, 1909, xxxvi, p. 285. 17 Silberberg : Ibid., 1909, xxxvi, p. 480. 18 Ehrenberg: Landw. Vers. Stat., 1910, Ixxii, p. 15. 19 Stoklasa : Compt. rend., 1913, elvi, p. 153. 20 Russell and Smith : Jour. Agr. Sei., 1906, i, p. 444. 21 Stoklasa : Expt. Sta. Rec, 1912, xxvi, p. 225. 14 Influence of Ar senk in Solls [Oct. Both ammonification and nitrification were greatly stimulated by arsenic trisidfide. It is likely that the sulfur played some part in this result. Demolon-^ attributed much of the fertilizing action of sulfur to its action on bacteria. The results which Russell and Hutchinsons^ obtained with calcium sulfide are interesting in this connection, They found that after thirty days there were five times as many organisms in soil to which calcium sulfide had been added as in the untreated soil, and that the yield of ammonia and nitrates in this time was one third greater in the treated soil than in the un- treated soil. These results show considerable similarity to the data reported here for arsenic trisulfide ; and most of the results obtained in this study may be interpreted by their theory^^ — that the soil con- tains another group of organisms which are detrimental to bacteria. If this theory be accepted, we must conclude that the soil bacteria are much more resistant to arsenic than are these other organisms, and that the soil organisms are able to function in the presence of very much larger quantities of arsenic than is this other class of organisms. For, while the antiseptics used by Russell and Hutch- inson in their experiments were subsequently removed, this was not the case with the arsenic in this work. As large a quantity of water-soluble arsenic was found in the soil at the end as at the beginning of an experiment. Some of the observed Stimulation was doubtless due to copper, lead, zinc, and sulfur, or whatever other foreign material was present in the insecticides. The con- stant ocurrence of a Stimulation in all the series points conclu- sively, however, to the fact that arsenic in some concentrations stim- ulates bacterial action. It must evidently occur in soil in large quan- tities before it becomes very toxic to the soil organisms. IV. SUMMARY One hundred parts per million of sodiiim arsenate may be ap- plied to a soil rieh in calcium and iron without materially decreasing the ammonifying or nitrifying powers of that soil. Smaller quan- tities may stimulate these activities. 22 Demoion : Compt. rend., 1913, elvi, p. 725. 23 Russell and Hutchinson : Jour. Agr. Sei., 1913, v, p. 173. 2* Russell and Hutchinson: Ibid., 1909, iii, p. m. 1913] J- E- Greaves 15 Zinc arsenite, lead arsenate and arsenic trisulfide stimulate the ammonifying activities of a soil, and their toxicity is not very marked until comparatively large quantities of arsenic are present, The two former reduce the ammonifying and nitrifying activities only one-half when 1,120 parts per million of arsenic are present, while arsenic trisulfide exerts a stimulating influence upon the am- monifying activities of the soil in the lower concentrations and does not become very toxic even in the highest concentrations. Paris green exerts marked toxicity on the ammonifiers, even "when present in small quantities. When present in large quantities it practically stops ammonification in soil. All these Compounds stimulated nitrification, the Stimulation being least for Paris green and greatest for lead arsenate. Arsenic trisulfide and Paris green, when present in large quan- tities, nearly stopped nitrification. Arsenic stimulated ammonification and nitrification, when it was present in soils in small quantities, but in very large quantities it was toxic. It is improbable, however, that lead arsenate, zinc arsenite, or arsenic trisulfide, will ever be applied to agricultural soil in quantities sußicient to become injurious to soil bacteria. Paris green may, but the quantity added would have to be large. The stimulating activity of the various Compounds added to the soil, upon ammonifying organisms and especially upon the nitrifying forms, is partly due to the anion and partly to the cation. Much of their action may be due to their influence upon injurious species. Water-soluble arsenic may exist as such in soils to the extent of 82 parts per million without entirely stopping ammonification and nitrification. Large quantities of ammonia and nitric nitrogen may be produced in a soil containing 50 parts per million of water- soluble arsenic, which is a greater quantity than any ever found in an agricultural soil. Measured in terms of their influence upon ammonification and nitrification as it takes place in soil, the toxicity of lead arsenate is the least. Next come zinc arsenite and arsenic trisulfide. The greatest toxicity is exerted by Paris green. From the results re- ported in the literature on the subject, this seems to be the sequence of toxicity when tests are made on the higher plants. i6 Influence of Arscnic in Solls [Oct, There is nothing in these results to indicate that arsenic trisulfide, or zinc arsenite, is as safe as lead arsenate for use as an insecticide. Arsenic trisulfide may be safer when first added to the soil. as is shown by its being almost insoluble when first applied and having practically no toxic influence upon ammonification; but, as bacterial action takes place in the soil, the arsenic of the arsenic trisulfide is much more soluble than that of lead arsenate, and becomes toxic to the nitrifying organisms when it is present in large quantities. THE NATURE OF HUMUS AND ITS RELATION TO PLANT LIFE* S. L. JODIDI {Office of Physiological and Fermentation Investigations, Bureau of Plant Industry, U. S. Department of Agriculture, Washington, D. C.) Considerable progress, recently, in connection with our knowl- edge of the chemical nature of "humus" makes it desirable briefly to review the results obtained. Up to a few years ago the generally accepted idea was that humus consisted of but a few organic Compounds. This was largely due to the work of a number of investigators, such as BerzeHus/ Detmer,^ Braconnot,^ Malaguti,^ Terreil,^ and especially Mulder^ and his school, who held that humus consists of a few simple organic substances which are chiefly acid in their nature (or at least can be converted into acids by treatment with alkahes), and which are closely related to each other. Thus, according to Mulder, ulmic acid, the first product in the decomposition of organic matter, is gradually converted into humic acid, geic acid, apocrenic and crenic acids, in the order named, all of which consist of but three Clements : carbon, hydrogen, and oxygen. This conception of Mulder's and of contemporary writers may have been due, in part, to the facts that protein matters were then assumed to have a uniform" composition; that the known carbohy- * Published by permission of the Secretary of Agriculture. Based largely upon the work conducted during the last six years by the writer while he was connected with the Michigan and Iowa Agricultural Experiment Stations. Re- ported to the Biological Division of the American Chemical Society (see p. 89). 1 Berzelius: Lehrbuch d. Chemie, 3. Aufl., 8 (1839). 2Detmer: Landw. Versuchsstationen, 14 (1871). ^Braconnot: Ann. chim. phys., 12, 191 (1819). 4Malaguti: Ibid. [3] 54. 407 (1858); Ann. (Liebig), 17, 52 (1836). ^Terreil: Bul. de la soc. chim. [2] 44, 2 (1885). 6 Mulder: Ann. (Liebig), 36, 243 (1840) ; Chemie der Ackerkrume; Jour. f. pract. ehem., 21, 343 (1840). ■^ Kossei: Berichte d. d. ehem. Ges., 34, 3245 (1901). 17 i8 Natur c of Hiwiiis [Oct. drates and their disintegration products were comparatively few (which was also true of the protein products) ; and last, but not least, that the methods of research in organic and biological chem- istry were inadeqnate. Recent investigations have thrown enough light upon the chem- ical natiire of humus or hiimus organic matter in the soil to demon- strate that it is a very complex material, which, in addition to dark colored hiimin substances, contains a large number of organic Com- pounds displaying acid, basic, neutral and amphoteric characters. The development of the idea of the chemical nature of humus Stands in a certain relation to the development of the chemistry of carbohydrates and proteins out of which humus is formed in the soil. Of the carbohydrates, mankind for centuries knew only cane sugar, which was originally obtained exclusively from the sugar cane. The same sugar was later discovered in the sugar beet (Marggraf, 1747), sorghum, maple tree and other plants. In addi- tion to this sugar there were discovered, also, lactose (Bartoletti, 1615), glucose (Lowitz, 1792), fructose (Dubrunfaut, 1847), ^^^^ so forth. At present we know a considerable number of sugars in the form of bioses, trioses, tetroses, etc., up to nonoses, i. e., sugars which contain in their molecules from two to nine carbon atoms, respectively, and which occur, in part as such, in nature ; the pentoses and hexoses and the corresponding Polysaccharides being the most important. As was demonstrated by many researchers, such sugars and, generally speaking, carbohydrates,^ when treated with acids or alka- lies, yield brown or black humin substances, whose physical and chemicarproperties remind one of soil humus to such a degree, that «Malaguti: Ann. (Liebig), 17, 52 (1836) ; Berzelius, Lehrbuch d. Chemie, 3. Aufl. 8 (1839) ; Conrad and Guthzeit, Ber. d. d. ehem. Ges., 18, 439 (1885) ; 19, 2850 (1886) ; Sestini, Gas. ehim. iL, 10, 121, 355. Grote and Tollens, Ann. (Liebig), 176, 181 (1875) ; 202, 226 (1880) ; Peligot, Ann. chim. phys. [2], 73, 208; Mulder, Ann. (Liebig), 36, 243 (1840); Chemie der Ackerkrume; O. Schmiedeberg: Arch. expt. Path. und PharmakoL, 39, l (1897); Hoppe-Seyler : Zeit. f. physiol. Chetn., 13, 66 (1889); Samuely: Beitr. ehem. Physiol. und Path., 2, 355 (1902). 1913] «S". L. Jodidi 19 the artificial and natural products were considered by some as closely related,^ by others even as identical. When, however, dilute acids are applied, the carbohydrates yield a number of well defined intermediary products. Thus, the Poly- saccharides furnish first monosaccharides, these latter yield organic acids, etc. For raffinose, e. g., we have: Raffinose (melitriose)-» melibiose (+ J-fructose)-»(i-glucose + galactose. The resulting monosaccharides can yield, e. g., lactic acid, butyric acid, alcohol, citric acid, etc., depending upon the conditions of transformation. So far as proteins are concerned, it was first thought that they have a uniform composition and constant properties. Modern re- searches revealed the fact that the various proteins have different chemical composition and structure. Their gradual decomposition leads (through the stages of proteoses and peptones) chiefly to diamino and monoamino acids. And it is the latter Compounds particularly that play a role in the formation of humin substances. A number of investigators^° have found that proteins when treated with acids yield humus-like substances. A further study of this phenomenon showed that it is particularly the diamino acid lysin,^^ and the monoamino acids tryptophan^^ and ty rosin (and glucosamin) that participate in the production of the melanoidins.^^ Hence, it is evident that plants containing proteins rieh in tyrosin, tryptophan, lysin and glucosamin radicals will, everything eise being equal, yield more humus than plants poor in those Compounds. When pure proteins are subjected to the influence of enzymes or to the activity of microorganisms, they are first hydrolyzed, chiefly to diamino acids and monoamino acids. The resulting pri- mary amino acids are, especially under the influence of microbes, subjected to secondary changes which lead to the formation of humin substances, f atty and hydroxy acids, phenols, basic substances, ^Sostegni: Landw. Vers.-Stat., 32, 9 (1885); Andre: Bull. soc. chim. [3] 21, 497 (1899) ; Eggertz: Chem. Centralbl., 343 (1889). 10 Mulder: Jour. f. pract. Chem., 21, 343 (1840); The Chemistry of Vegetable and Animal Physiology, trans. by Fromberg (Edinburgh and London), 1849, p. 153; Schmiedeberg: Arch.f.exper.Pathol.u.Pharmakol, 39,65 (1897); Panzer: Zeit. f. physiol. Chem., 33, 131 (1901). 11 Hart: Zeit. f. physiol. Chem., 33, 355 (1901). 12 Hopkins and Cole: Journ. of Physiol., 27, 418 (1901) ; 2g, 451 (1903). ^^Samuely: Hofmeiste/s Beiträge, 2, 355 (1902). i- 20 Nature of Humus [Oct. etc. Thus, the Splitting off of carbon dioxid from the diamino acids as well as of the NHj group from the monoamino acids, leads to the formation of amins and acids respectively, as illustrated by the following equations : NH2-CH2-CH2-CH2-CH2-CH-NH2-COOH -^ CO2 + NH2-CH2-CH2-CH2-CH2-CH2-NH2 Lysin Cadaverin CH CH CH^\C C-CH,-CH-NH,-COOH CH^\C C-CH.-CH^-COOH CH CH ^'H CH NH CH Tryptophan Indolpropionic acid In the case of nucleoprotelns there result, in addition to protein products, purin derivatives and pyrimidin bases. When carbohydrates are subjected to the influence of enzymes and bacteria, they also, as the result of primary hydrolysis, are first split into smaller carbohydrate molecules, which then may be further decomposed to alcohol, organic acids, etc. In the same manner, f ats, under the influence of similar agencies, are split into fatty acids and glycerol. There is no reason to assume that the proteins, nucleoprotelns, carbohydrates and fats in vegetable and animal remains, when sub- jected to the action of such agencies in soil, would not yield decom- position products similar to those that result in the case of pure proteins, nucleoprotelns, carbohydrates and fats. Recent investigations show that humus, using this word in its widest sense, is very complex, consisting of a large number of Com- pounds. Contrary to the findings of earlier writers (see p. 17) humus contains, in addition to carbon, hydrogen and oxygen, also nitrogen, sulfur and phosphorus. It contains acids of well known composition and Constitution, e. g., monohydroxystearic,^^ dihydrox- ystearic, oxalic, succinic, acrylic, saccharic. It contains basic sub- 1* Schreiner and Shorey: Bul. Nos. 53 and 74, Bureau of Solls, U. S. Dept. Agr.; Jour. Amer. Chem. Soc, 32, 1674 (1910) ; Proc. Eighth Intern. Congr. Appl. Chem., 15, 247 (1912). 1913] S. L. Jodidi 21 stances, ^. ^r., diamino acids (Kossel's hexon bases)/^ purin^^ bases, and amins.^^ It contains a variety of "neutral" Compounds, e. g., hydrocarbons/^ esters, aldehydes, as well as amphoteric substances like amino acids,^^ etc.^° Moreover, there is very little doubt that the number of definite organic Compounds which can be extracted from " soil organic matter " will be considerably increased within the next few years. Mention may be made here of the fact that, of the Compounds found in " soil organic matter," the amino acids and acid amides play a prominent role, for the reason that they are contained in pre- dominant proportions in acid extracts of soils ; and for the further reason that they represent an important source for the production in the soil of ammonia,^^ and hence of nitrates. Considering that certain constituents (the ten well known Cle- ments) are absolutely indispensable for plant life, it is easy to un- derstand why humus is called by many the "life of the soil." Not only does it contain most of the elements which are necessary for plant life, like nitrogen, phosphorus, sulfur, etc., but, what is of equally great importance, it affords a means for rendering more of the necessary inorganic elements available. The carbon dioxide, nitric acid and sulfuric acid which result through oxidation of the elements carbon, nitrogen and sulfur in humus, are powerful agents for the extraction of indispensable ele- ments (potassium, calcium, magnesium and others) from the rocks, thus Converting rocks and rocky land into fertile arable soil. 15 Jodidi: Techn. Bul. No. 4 {1909), Mich. Agr. Expt. Station; Research Bul. No. I, Iowa Agr. Expt. Sta. (1911) ; Jodidi and Wells: Research Bul. No. 2, Iowa Agr. Expt. Sta. (1911). 18 Schreiner and Shorey: Jour. Biol. Chem., 8, 385 (1910). 1'' Shorey: Proc. Eighth Intern. Congr. Appl. Chem., 15, 249 (1912). 18 Shorey: Proc. Eighth Intern. Congr. Appl. Chem., 15, 248 (1912). lö Jodidi: Jour. Amer. Chem. Soc, 32, 396 (1910) ; 33, 1226 (1911) ; 34, 94 (1912); Schreiner and Shorey: Jour. Biol. Chem., 8, 381 (1910) ; Robinson: Techn. Bul. No. 7, Mich. Agr. Expt. Sta. (1911) ; Jour. Amer. Chem. Soc, 33, 564 (1911). 20 A good account of the Compounds extracted from soils is contained in Proc. Eighth Intern. Congr. Appl. Chem., 15, 248-250 (1912). 21 Jodidi, Kellogg and Snyder: Research Bul. No. g, Iowa Agr. Expt. Sta- tion (1912) ; Jodidi: Jour. of the Franklin Institute, 175, 245 (1913) ; Ibid., 175, 483 (1913) ; Proc. Eighth Intern. Congr. Appl. Chem., 26, 119 (1912), 22 Natur e of Humus [Oct. It is well known that humus improves the physical condition of the soil. It increases, for instance, a soil's capacity to hold water, to retain valuable nitrogenous constituents, to resist corrosion. It binds the particles of sandy soils. It makes clayey soils friable, increasing at the same time their capacity to absorb the sun's raya and rendering the soil-temperature more uniform. In other words, humus makes the soil a more habitable and suitable home for the Performance of the life functions of plants. Thus, we find the humification process inserted into the chaln of nature's cycles as a necessary link, without which the perpetual con- tinuance of plant life cannot be conceived. Again, man's food, whether of vegetable or animal origin, is composed chiefly of pro- teins, fats, carbohydrates and mineral substances, all of which are contained in the plant and animal bodies. However, animal life is, in the last analysis, based upon the presence of plants, the digestion and assimilation of which give the material for the formation in the animal body of its organs and tissues. Again, the plants need for their life certain elements which are present in humus. Here we have, then, a cycle in which the physical life of man, as well as the existence of the animal and vegetable kingdoms, are brought into close connection with humus. So close is this relation thatconditions indispensable for life are also necessary for decomposition of humus materials; extremes that exclude life also render decomposition im- possible. There is practically no plant life in arctic regions or throughout the winter, and there is no decomposition under the same conditions. On the other hand, plant life is luxuriant in the tropics. There, too, decay is very rapid. The plant and the animal kingdoms need air for their life. The same air is indispensable for decomposition. Equally, neither plant life nor decomposition of organic substances is possible without water. In the process of humification, nature has a powerful means for the utilization of vast amounts of waste materials for purposes of life. Or to put it in other words: It is the humification of vege- table and animal remains that makes them available for new genera- tions of plants. CLEAVAGE OF BENZOYLALANINE AND ACETYL- GLYCINE BY MOLD ENZYMES* ARTHUR W. DOX and W. EUGENE RUTH (Chemical Section of the Iowa State College, Arnes, Iowa) The readiness with which certain aromatic acids, in which the carboxyl group is attached to a cydic nucleus, conjugate with gly- cine in the animal body is well known. Among the substances which undergo this synthesis the most familiär is benzoic acid and its Substitution products. The reaction is, however, not limited to homocyclic Compounds, since both five and six membered heterocyc- lic derivatives, among which may be mentioned pyromucic, thio- phenic and pyridinic acids, also unite with glycine. A reversal of this reaction takes place under the influence of certain enzymes. Such enzymes occur in plants as well as in animal tissues. One of us, in collaboration with Neidig, has shown that enzyme preparations from some of the lov/er fungi can bring about the hydrolysis of hippuric acid^ and also that of pyromucuric acid.^ It was thought possible, however, that the enzymic hydrolysis might be of wider application than the corresponding synthesis. In the animal organism the best known conjugations are those in which the substance to be excreted is united with glycine, glucuronic acid, sulfuric acid or cysteine. Homologues of glycine do not enter into synthetic reactions with the formation of excretory products, nor do acid radicals of the aliphatic series unite with glycine. Consider- ing the diversity of aromatic radicals capable of combining in this way with glycine and not with the other amino acids, the question arises as to what determines the specificity of the reaction, and whether the corresponding hydrolysis would be equally specific. For instance, is the cleavage of such Compounds dependent upon the ♦Read at the Rochester meeting of the American Chemical Society, Sep. 10, 1913. (Page 83.) ^Dox and Neidig: Zeitschr. f. physiol. Chem., 1913, Ixxxv, p. 68. 2 Dox and Neidig : Biochem. Bull., 1913, ii, p. 407. 23 24 Cleavage of Benzoylalanine and Acetylglycine [Oct. presence of the glycine group or the presence of an aromatic acid radical? Or does hippuric acid simply represent a type of substi- tuted amino acids, all of which are hydrolyzed by the same enzyme? With the view of throwing some light on this question, ben- zoylalanine and acetylglycine were subjected to the action of en- zyme preparations capable of hydrolyzing hippuric acid. Benzoyl- alanine may be regarded simply as a homologue of hippuric acid. Neither this substance nor acetylglycine, as far as \ve are aware, occurs in nature nor does either result from any known biological process. Mold cultures were made as previously described^ and the press Juice allowed to act upon the sodium salt of benzoylalanine and acetylglycine respectively. Since the experiments were conducted exactly as outlined in our previous paper, the details will not be re- peated here. The formol titration was made after the enzyme and Substrate had been in contact for two weeks at room temperature. The results are given in the accompanying tables. TABLE I Data on the cleavage of hensoylalanine Source of enzyme Titration : «/ioBa(OH)2, c.c. Control : «/ioBa(OH)2, c.c. Difference, c.c. Cleavage, As-bereillus nieer 7.41 13.17 8.46 12.95 10.66 11.31 9-96 4-45 10.00 5.28 10.14 8.09 9-65 8.13 2.96 3-17 3-18 2.81 2-57 1.66 1.83 22 8 Aspergillus clavatus 24.4 24-5 21.6 19.8 12.8 Aspergillus fumigatus Cladosporium herbarum Fusarium oxysporium Penicillium roqueforti Penicillium expansum 14.1 TABLE 2 Data on the cleavage of acetylglycine Source of enzyme Titration : «/ioBa(0H)2, c.c. Control : «/ioBa(OH)2, c.c. Difference, c.c. Cleavage, As-bereillus nieer 28.92 20.05 10.79 1-38 22.56 13-73 25-33 10.62 6-79 2.78 1.76 6-43 2.03 6.53 18.30 13.26 8.01 86.93 62.99 38.09 none Aspergillus clavatus As-bereillus fumieatus Cladosporium herbarum Fusarium, oxvs-öoriufn 16.13 11.70 18.80 76.62 55-58 89-31 Penicillium roqueforti Penicillium expansum 3 Dox and Neidig: Loc. cit. 1913] Arthur W. Dox and W. Eugene Ruth 25 Both benzoylalanine and acetylglycine are hydrolyzed by an en- zyme present in lower fungi. In the case of benzoylalanine the cleavage is considerably less than in that of acetylglycine. A possi- ble explanation of this difference may be the fact that the racemic mixture was employed and the enzyme was specific for only one isomer. This, however, cannot be stated with any certainty since optical studies were not made. It appears probable from the above results that the enzymic cleavage of substituted amino acids is not limited to Compounds strictly analogous to hippuric acid, which occur as excretory products. The reaction is therefore not specific for glycine derivatives, nor for benzoyl or similar radicals contain- ing a cyclic nucleus. A COLOR REACTION OF GLYCINE WHEN BOILED WITH CHLORAL HYDRATE EDWIN D. WATKINS (University of Tennessee, Memphis) Considering the color reaction of triketohydrinden hydrate when boiled with amino acids, I was led to investigate the possible color reactions of other substances supposed to have two — OH groups attached to one carbon atom, when boiled with Solution of amino acid. Only glycine has been studied. This report is a prelimi- nary one. The materials used in the experiments were the purest procur- able in the market. An aqueous Solution of glycine in a beaker was treated with chloral hydrate in substance and boiled for five minutes. The Solu- tion assumed a dark red color, which was very marked. Aqueous Solutions of glycine and chloral hydrate boiled separately remained colorless. Glycine in dilute aqueous Solution (i to 5,000), treated in this way, yielded a distinct dark red color. A weaker Solution of gly- cine (i to 10,000) yielded faint amethyst color. Phenol, glycerol, resorcin, acetone, ethyl alcohol, glyoxylic acid, orthophosphoric acid, and chloral itself, when boiled with aqueous Solution of glycine, yielded no color. Acetone boiled with barium hydroxid sol. and then with glycine sol., yielded a green color which changed to dark red in thirty minutes. A cold saturated Solution of glycine in water was treated with chloral hydrate in substance and boiled to half the original volume. A port-wine colored Solution resulted which, when treated with ether and extracted, yielded its color in part to the ether. The ether Solution was evaporated over an incandescent lamp; there resulted a dark oily liquid which gave off pungent fumes. This oily liquid, when cooled rapidly on a glass slide with solidi- 26 1913] Edwin D. Watkins 27 fied carbon dioxide, showed microscopic needles and leaf crystals which rapidly llquefied. The oily liquid yielded a green Solution with strong hydrochloric acid. This Solution was evaporated over a water bath. Portions of the syrupy fluid remaining just before dryness were placed on a slide with a platinum loop and allowed to cool. There formed microscopic crystals of brown leaves and balls. A control was run by boiling chloral hydrate and extracting with ether. The ether Solution was evaporated over an incandescent lamp, when typical chloral hydrate leaves and clumps crystal- lized out. The nature of the various products has not been determined, They are now under investigation. STUDIES ON WATER DRINKING 15. The Output of fecal bacteria as influenced by the drinking of distilled water at meal time N. R. BLATHERWICK AND P. B. HAWK (Laboratories of Physiological Chemistry of the Jefferson Medical College and the University of Illinois) Introduction. The first study of bacterial growth under the influence of water-ingestion was made by Fowler and Hawk (i), and somewhat later a more extended study of the problem was made by Mattill and Hawk (2). In the latter investigation it was found that the Ingestion of large amounts (1,000 c.c.) of water with meals caused the protein constituents of the food to be more fully utilized as shown by a decrease in all forms of nitrogen in the feces, including bacterial nitrogen. When 500 c.c. of water were taken with meals no significant changes in protein utilization were evident. However, the data admitted of the negative conclusion that no undesirable efTects followed the water-ingestion. In each of the experiments mentioned, softened water (2^) had been employed. The beneficial influence exerted by the drinking of softened water at meal time having been indicated by these ex- perimenters, the question naturally arose whether the drinking of distilled water at meal time would have a similar influence. It has been quite generally supposed that distilled water, on account of its lack of salts, would consequently have an untoward influence on the processes of digestion and absorption. So far as we know, no experiment to prove the truth or falsity of this belief has ever been performed. Findlay (3) wTOte as follows concerning the influence of dis- tilled water upon the tissues. "If tissues or cells are placed in dis- tilled water, passage of water into the cells occurs owing to the difference of osmotic pressure. The cells swell up and may finally burst and die. A similar poisonous action on cells is observed when 1913I N' R' Blatherwick and P. B. Hawk 29 distilled water is drunk. In this case the surface layers of the epithelium of the stomach undergo considerable swelling; salts also pass out, and the cells may die and be cast off. This may lead to catarrh of the stomach. It is to this action of pure water that the harmful effects of melted snow or ice is due, since freezing puri- fies the water. For this reason also, one of the Springs of Gastein has come to be known as the Poison Spring, although its water is purer than ordinary distilled water." If Findlay's argument regarding the pernicious influence of dis- tilled-water Ingestion is true, a very decided exception was noted in one of the fasting experiments made at the University of Illinois. In this instance a dog was fasted 117 days (4) and received, by means of a stomach tube, a daily ration of 700 o.e. of distilled water. The animal was then fed and brought back to normal weight, and again fasted for 104 days. At the end of this ex- tremely long period of inanition, the organs and tissues of the animal were carefully examined. No signs of a deranged gastric mucosa were in evidence. If the toxic influence of distilled water is as pro- nounced as Findlay would have us believe, certainly a period of 117 days is a sufficiently long interval in which to demonstrate such an influence. This would appear to be particularly true in the case of a fasting animal, whose resistance to such toxic influence may have been lowered somewhat. Granting the validity of Findlay's claim, his contention cannot be advanced as evidence of the harmful influence of drinking dis- tilled water with meals. Because of the electrolyte content of the average diet, the distilled water would cease to act as distilled water soon after its entrance into the stomach. If distilled water is to be considered as having a toxic influence upon the gastric mucosa, such toxic effect must of necessity be more pronounced when the distilled water is introduced into an empty stomach. It will be apparent from the discussion which follows that we were able to detect no harmful influence exerted by the distilled water in the experiments herewith described. Koeppe (5) has voiced the opinion that the catarrh of the stom- ach which may follow the excessive Ingestion of ice arises because of the lack of salts in the water from the melting ice. Nocht (6) 30 Studies on Water Drinking [Oct. and Winkler (7), on the contrary, cite several cases in which pro- longed, distilled-water ingestion was unaccompanied by any harm- ful influences. Harlow (8) considers that the ingestion of water from glaciers leads to irritation of the mucosa of the gastrointestinal tract. Spitta (9) has recently expressed the opinion that the ques- tion whether distilled-water ingestion is harmfnl mtist be considered an open one. Very recently Oehler (10) has ptiblished data from a Short series of tests upon white mice from which he draws the conclusion that distilled-water ingestion by these animals causes hemoglobinuria. The introdtiction of distilled water into the cir- cidation will, of course, be followed by a transient hemoglobinuria. It is a little difficult to see, however, how the introduction of the fluid mto the stomach can bring about such a condition. We expect to investigate the question shortly. Incomplete digestion and absorption of the protein part of the food causes an increased elimination of nitrogen in the feces, or may result in an increased grow^th of bacteria in the lower intestine. Hence any experiment to show the course of bacterial development may have a direct bearing on the problems of digestion and absorp- tion. It may be argued in a general w^ay, for example, that an increase of bacteria shows a decreased digestive or absorptive effi- ciency, and a decreased bacterial content of the feces indicates a more efficient functioning of the organs of digestion and absorption. Description of the experiments. Methods. Fresh samples of feces w^ere used for all analyses. Duplicate analyses were made in all cases, approximately 2 gm. of fecal matter being used in each determination. Total nitrogen was determined by the Kjeldahl method. The method used for bacterial nitrogen was in general the modification of MacNeal's method described by Mattill and Hawk (11). It differed in some of the details from the directions given by these workers. Two grams of fresh feces were weighed by difference into a 50 c.c. centrifuge-tube, rubbed up with 0.2 percent hydrochloric acid sol., and the bacterial matter brought into Sus- pension in the usual manner. When the first serial centrifugation was completed, about one-half the Suspension was transferred to a 100 c.c. centrifuge-tube. The experiment was then continued, using the larger tubes throughout the remainder of the experiment. The 1913] N- R- Blatherwick and P. B. Hawk 31 use of the larger tubes shortens the time of the Sedimentation, at the same time making as good a Separation as is procured by the use of the smaller tubes. Another point of difference was the manner of transferring the bacterial substance to the Kjeldahl flasks after the final Sedimentation. It was found, after some experimentation, that the bacterial substance settled more completely to the bottom of the centrifuge tube when the bacteria were suspended in sulphuric ether instead of ethyl alcohol. In all these analyses the sediment from the alcohol was therefore centrifuged with ether, the sediment being transferred to the Kjeldahl flask and the nitrogen determined in the usual way. The centrifuge used in this experiment was run by an electric motor, which developed a speed of 1,400 revolutions per minute. It will be noted here that this is a slower rate of speed than is gener- ally used. More will be said of this in a later connection. SuBjECTS, PLAN, DiET, ETC. Two studcuts, normal in every respect, were given a uniform diet for several days or until nitro- gen equilibrium was reached, as shown by analyses of foods and excreta. During this period (preliminary), 100 c.c. of distilled water were taken with each meal. Besides this, 200 c.c. of distilled water were ingested by subject V at 10 A. M., at 3 P. M., and at 8.30 P. M. Subject C ingested a total of 400 c.c. at these hours. After the subjects had reached nitrogen equilibrium, they were placed on the " moderate water " diet, which differed from the pre- liminary only by the addition of 500 c.c. of distilled water at each meal. This made for subject V a total water-ingestion of 900 c.c. during the preliminary period, and of 2,400 c.c. during the "moderate water" period. The values for subject C were 700 c.c. and 2,200 c.c. respectively. The duration of the "moderate water" period was ten days. The subjects were then returned for an interval of five days to the diet of the preliminary period, this period being called the intermediate period. The " copious water " period was then begun, and continued five days. The diet during this period differed from that of the preliminary and intermediate periods by the addition of 850 c.c. of distilled water at each of the three meals, making a total daily ingestion of 3,450 c.c. for sub- ject V, and of 3,250 c.c. for subject C. The diet of the preliminary 32 Studies on Water Drinking [Oct and intermediate periods was then resumed for five days, this con- stituting the final period. In all cases charcoal capsules were used to separate the feces of the different periods. Analyses were always made on fresh indi- vidual stools, unless the amount was too small, in which case it was placed in a refrigerator in an air-tight receptacle; and upon the following day was mixed with the succeeding stool and analysis made on the composite sample. Subject V was 24 years of age and weighed 58 kg. Subject C was 29 years of age and weighed 60 kg. The hours for meals were as follows: breakfast 7-7.30; dinner 12—12.30 and supper 6-6.30. The temperature of the water ingested by subject V was 14° C. whereas that ingested by subject C was 22° C. The diet was the same for each subject and consisted of the fol- lowing constituents, which were fed at each of the three daily meals : Graham crackers, 100 gm.; peanut butter, 15 gm.; butter, 25 gm.; milk 400 c.c. Discussion of results. Moderate water-drinking. Subject C. The decrease of fecal nitrogen from 1.236 gm. in the pre- liminary period (see Table i) to 0.943 gm. in the "moderate water" period, with the corresponding drop in bacterial nitrogen from 0.729 gm. to 0.544 gm., was unexpected, since previous experi- ments in this laboratory (2) showed moderate water-drinking to have very little effect on the fecal-nitrogen excretion. In fact, the alterations in this excretion were heretofore so small that it was impossible to draw any positive conclusions. In this case, however, we have clear evidence to show that the processes of digestion and absorption have been improved by the drinking of an additional 1,500 c.c. of distilled water at meal time. The beneficial efifects were not confined to the water period, but were carried over into the intermediate period. Although there was but 0.047 gm. decrease in bacterial nitrogen from the "moderate water" period to the intermediate period, the percent of nitrogen occurring as bacterial nitrogen was decreased from 57.67 percent to 53.78 percent. This confirms the previous findings that the influence of the "high water" ingestion was not confined alone to the interval during which it was being ingested. I9I3] N. R. Blatherwick and P. B. Hawk 33 TABLE I Subject C Preliminary Period. Number of stool Weight of stool, grams Dry matter, per Cent. Amount of dry matter, grams Fecal nitro- gen, grams Bac- terial ni- trogen, grams Bacterial dry sub- stance (cal- culated),* grams Dry bacteria in dry feces (calcu- lated),* per Cent. Bacterial nitrogen in fecal nitrogen, per Cent. I 2 3 4 5 6 41.0 103.0 83.0 60.5 179-5 53-5 27.85 27.86 26.40 25-77 26.68 25-77 11.42 28.70 21.91 15.60 47-90 13-78 0.819 1.911 1.079 0.737 2.158 0.713 0-505 1.184 0.634 0.422 1.219 0.413 4.608 10.803 5-785 3-854 II. 122 3-768 40-35 37-64 26.40 24.68 23.22 27-34 61.66 61.96 58.76 57.26 56.49 57-92 Total 520.5 139-31 7-417 4-377 39-936 Average 86.75 26.76 23.22 1.236 0.729 6.656 28.67 59.01 "Moderate Water' • Period I 103-5 23-85 24.70 1-339 0.769 7-0x6 28.40 57-43 2 65-5 25-11 16.4s 0.830 0.528 4.817 29.28 63-61 3 130-0 24.60 32.00 1.872 1.004 9.160 28.63 53-63 4 44-5 22.95 10.21 0.617 0.397 3-622 35-48 64-34 S 70-5 23-86 16.82 0.907 0.545 4-973 29-57 60.09 6 56.0 22.37 12.53 0.750 0.442 4-033 32.19 58.93 7 83-5 26.16 21.84 1.223 0.620 5-657 25.90 50.70 8 50.0 26.61 13-30 0.664 0.378 3-449 25-93 56.93 9 97-0 27-35 26.53 1.227 0.755 6.889 25-97 61.53 Total 700.5 174-38 9.429 5-438 49.616 Average 70.1 24.89 17.44 0.943 0.544 4.962 28.44 57.67 Intermediate Period. z 43-5 23.42 10.19 0.535 0.299 2.728 26.77 55-89 2 43-5 26.68 11.60 0.612 0.311 2.838 24.47 50.82 3 60.5 30.37 18.37 0.779 0.421 3-841 20.91 54-04 4 73-5 29-75 21.87 0.933 0.523 4-772 21.82 56.06 S 136.0 27.69 37.66 1.760 0.930 8.485 22.53 52-84 Total 357-0 99-69 4.619 2.484 22.664 Average 71.4 27.92 19.94 0.924 0.497 4.533 22.74 53.78 "Copious Water" Period I 57-5 23-77 13-67 0.800 0.413 3-768 25-56 51-63 2 43-5 29.67 12.91 0.644 0.344 3-139 24.31 53-42 3 56.5 25.60 14.46 0.784 0.423 3-859 26.69 53-95 4 86.5 28.25 24-44 1-153 0.666 6.077 24.86 57-76 5 93-5 28.72 26.85 1.240 0.687 6.268 23-34 55-40 Total 337-5 92.33 4.621 2.533 23.111 Average 67.5 27.34 18.47 0.924 0.507 4.622 25.03 54.81 Final Period. I 2 3 4 5 72.0 66.5 72.5 74-5 72-5 27.20 27.17 25-18 23.20 22.68 19-58 18.079 18.25 17.28 16.44 0.998 0.941 0.937 0.948 l.OOI 0.536 0.541 0.540 0.539 0.571 4.890 4-845 4.927 4.918 5-210 24-97 25-81 26.00 28.46 31.69 53-71 55-95 57-63 56.86 57-04 Total 358.0 89.62 4-833 2.717 24.790 Average 71.6 25.03 17.92 0.967 0.543 4.958 27.66 56.22 * Previous tests showed that dry bacteria contain 10.96 percent of nitrogen. 34 Studies on Water Drinking [Oct. Siihject V. The differences here are not so great as in the case of subject C, but it will be seen, by examining Table 2, that the values for the intermediate period are in all cases less than for the preliminary period, Copious WATER-DRiNKiNG. It will be recalled that this experi- ment was a continuation of the former, the intermediate period serving as the final period for the "moderate water" experiment, and as the preliminary period for the experiment on copious water- drinking at meal time. As was mentioned before, the "copious water" period differed from the intermediate and final periods in that 850 c.c. of distilled water were added to the water-ingestion of each of the daily meals. This made a total water-ingestion of 3,450 and 3,250 c.c. per day, for subject C and for subject V, respectively. Siihject C. It is interesting to note that the values for total nitrogen, bacterial nitrogen and percent of bacterial nitrogen in fecal nitrogen, were less in all cases in the "copious water" period than in the "moderate water" period; also, that the values for the above were less in the final period than they were in the preliminary period. This would seem to indicate clearly that the drinking of moderate quantities of water with meals, 2,400 c.c. daily, had a beneficial influence, while the beneficial influence of larger quantities, 3,250 c.c. daily, was still more pronounced. The most significant point regarding the bacterial-nitrogen excretion of subject C is the fact that the value for the preliminary period was much higher than that for any one of the four periods which followed. In other words the daily Ingestion of 1,500 c.c. of water during the period of moderate water-ingestion caused a very pronounced reduction in the growth of intestinal bacteria ; and this lowered development continued throughout the remainder of the experiment. The daily output of dry bacterial substance for the preliminary period (6.65 gm.) when compared with similar values for the other periods of the experiment (4.96, 4.53, 4.62, and 4.95 gm.) demonstrates this point very nicely. Siihject V. Another interesting comparison may here be made between the two subjects. As with subject C, the values for daily excretion of total fecal nitrogen, bacterial nitrogen and percent of I9I3] N. R. Blatherwick and P. B. Hawk 35 TABLE 2 Subject V Preliminary Period. Bac- Bacterial Dry bacteria Bac- terial Dry Amount of Fecal terial dry sub- in dry nitrogen Number Weight of matter. dry matter. nitrogen, nitro- stance (cal- feces in fecal of stool stool, grams per Cent. grams grams gen, grams culated),* grams (calcu- lated),* per Cent. nitro- gen, per Cent. I 51-5 21.25 10.94 0.842 0.466 4.247 38.82 55-32 2 189.5 17.96 34-04 2.274 1.489 13-586 39-91 65-47 3 198.5 17.42 34-58 2.370 1-366 12.463 36-04 57-65 4 75-0 16.64 12.48 0.887 0.574 5-234 41.94 64.68 5 86.0 19-54 16.80 1.069 0.687 5-269 37-32 64.27 6 85.0 21.11 17-94 1.056 0.635 5-797 32.31 60.17 7 84-S 21.90 18.50 1.072 0.609 5-558 30.04 56.83 8 240.0 17-32 41-57 2.530 1.364 11-445 29-94 53-91 Total 1,010.0 186.85 12.098 7.190 65-599 A verage 126.3 18.50 23.3s I.512 0.899 8.200 35.11 59-43 'Moderate Water" Period. I 92.5 12.98 12.01 0.868 0.395 3.608 30.04 45-57 2 102.5 16.23 16.63 1-137 0.721 6.579 39-56 63.42 3 101.5 20.42 20.72 1.202 0.738 6.737 32.51 61.43 4 153-5 17.61 27-03 1.710 i.iii 10.137 37-50 64-97 5 2350 12.50 29-37 2-341 1-349 12.308 41.91 57-62 6 115. 5 15-14 17-49 1.288 0-754 6.876 39-31 58.51 7 93-5 20.02 18.72 1.080 0.822 7-503 40.08 76.14 8 III. 18.15 20.14 1.342 0.856 7-813 38-79 63.81 9 92.5 21.13 19-54 I-152 0.679 6.194 31-70 58-93 IG 105.0 19-39 20.34 1-305 0.789 7.210 35-45 60.47 Total 1.202.5 201.99 13-425 1.343 8.215 74-965 A verage 120.3 16.80 20.20 0.822 7.497 37.11 61.19 Intermediate Period. I 48.5 20.98 10.17 0.625 0.358 3.270 32.15 57-37 2 56.5 24.17 13-65 0.803 0.463 4.224 30.95 57-75 3 244-5 19.26 47-09 2.698 1.639 14-954 31-76 60.74 4 154-5 17.60 27.19 1.601 1.014 9-252 34.02 6333 5 115-5 19-25 22.23 1-515 0.900 8.209 36.93 59-39 Total 619-5 120.33 7.241 4-374 39.909 Average 123.9 19.42 24.07 1.448 0.875 7-982 33-17 60.41 'Copious Water" Period. I 56.0 19.02 10.65 0.616 0.383 3-496 32.83 62.22 2 75-5 22.39 16.90 X.084 0.588 5-366 31-75 54-25 3 44-5 15-84 7-05 0.625 0.360 3.288 46.64 57.66 4 120.0 22.29 26.75 1.500 0.856 7.812 29.20 57.08 5 171-S 22.51 38.60 2.024 1.189 10.848 28.10 58.75 Total 467-5 99-95 S.849 3.376 30.810 Average 93.5 21.38 19.99 1.170 0.675 6.162 30.83 57.72 Final Period. I 2 3 4 162.5 124.5 56.5 214.0 20.29 19.19 21.07 16.13 32.97 23.89 11.90 34-52 1.890 1-432 0.714 2.222 1.004 0.781 0.420 1.258 9.160 7.122 3.835 11.478 27.78 29.81 32.22 33.25 53-12 54-51 58-87 56.61 Total 557.5 103.28 6.258 3.463 31.595 Average 111.5 18.53 20.66 1.252 0.693 6.319 30.59 55.34 * See note, bottom of Table i. 36 Studies on Water Drinking [Oct. bacterial nitrogen in fecal nitrogen, were less in the period of copiotis water-ingestion than in the "moderate water" period. Also, the values for these forms of nitrogen were less in the final period than in the preliminary period. Thus, data obtained from two subjects, while differing in detail, show the same general fea- tures, moderate amounts of water producing desirable results, while larger amounts have an augmented effect for the better. General discussion. Bacteria-excretion. As Ehrenpfordt (13) has shown, the different values for bacterial nitrogen are caused by a diversity in the centrifugation procedure. With a high rate of speed the lighter bacterial substances are thrown out of Sus- pension, while a slower rate of speed will cause more of these par- ticles to remain in Suspension during the course of a given length of time. Thus, it would appear to be almost impossible to stop at just the moment when all the non-bacterial substances have been removed and all of the bacteria are still in Suspension. Ehrenpfordt has further stated that entirely comparable and trustworthy results are obtained by any given worker, using the same technic throughout. The absolute values are not so important as are the relations be- tween values of different parts of an experiment obtained by a single experimenter. Harris (12) as the result of recent experiments wrote as fol- lows regarding the centrifugation procedure for bacteria deter- mination: "I am of the opinion that if workers in this country, at least, are to continue using the method, some endeavor ought to be made to unify or standardize the technic; otherwise interlabora- tory results cannot be considered comparable." Siibjcct V. Thirty-two stools were examined and analyzed for fecal bacteria-nitrogen, the method described by Mattill and Hawk (11) being employed with modifications as stated above. The percent of dry bacteria in the dry feces was found to vary from 27.78 percent to 46.64 percent, with an average of 33.36 per- cent. The ratio of bacterial nitrogen to fecal nitrogen varied from 53.12 percent to 76.14 percent, the average being 58.82 percent. The average daily excretion of bacterial dry substance was 7.232 gm. Suhject C. Thirty stools were examined and produced the foUowing results: 26.51 percent of dry bacteria were found in the 1913] N- R- Blatherwick and P. B. Hawk 37 dry feces, The values varied from 20.91 percent to 40.35 percent. The average percentage of bacterial nitrogen was 56.25, the low value being 50.70 percent and the high value 64.34 percent, The amount of bacterial dry substance excreted daily was 5.146 gm. The above facts may be summarized as follows : Bacterial dry substance, g^ams Dry bacteria dry feces, per cent. Bacterial nitrogen in fecal nitrogen, per Cent. Subiect V 7.232 5.146 33.36 26.51 58.82 56.25 Subject C Average 6.189 29.94 57-54 Nutrition experimenters have obtained the following results for the amounts of dry bacteria excreted daily : Strasburger (14) 8.0 grams Sato (15) 8.54 grams Berger and Tsuchiya (16) 3.023 grams MacNeal, Latzer and Kerr (17) 5.34 grams Mattill and Hawk (2, 11) 8.27 grams Blatherwick and Hawk 6.189 grams Values obtained for the percentage of dry bacteria in dry feces are as follows: Strasburger (14) 24.3 percent Schittenhelm and Tollens ( 18) 42.0 percent Lissauer (19) 8.67 percent Harris (12) 9.18 percent Tobaya (20) 1 1.22 percent Sato (15) 24.39 percent Berger and Tsuchiya (16) 12.6 percent MacNeal, Latzer and Kerr (17) 26.9 percent Mattill and Hawk (2, 11) 27.95 percent Blatherwick and Hawk 29.94 percent It is interesting to note, here, that the values for bacterial dry matter and for percent of dry bacteria in dry feces obtained from subject C are almost identical with those reported by MacNeal, Latzer and Kerr. Their values were 5.34 gm. and 26.9 percent respectively, and ours 5.146 gm. and 26.51 percent. 38 Studics on Watcr Drinking [Oct. Since these analyses were all made at the same time, using the same centrifuge and the same technique, and uniformly higher re- sults were obtained in the case of subject V throughout, it would appear that no hard and fast value can be laid down for the daily excretion of fecal bacteria. The questions of individuality, diet, and speed of centrifugation are important factors in bringing about the final results. The relationship of urinary indican to fecal bacteria. Urinary indican is considered to be an index of intestinal putre- faction. If it is a true index then the relationship of the urinary indican excretion to the Output of fecal bacteria ought to be of inter- est. For this reason the indican content of the urines from the sub- jects of this investigation was determined (21). A modification of Ellinger's method was used. Table 3 summarizes the corresponding values for indican (mg.) and bacterial nitrogen (gm.). TABLE 3 Relation between urinary indican and fecal-bacteria nitrogen Experimental period Constituent determined Preliminary " Moderate water" Inter- mediate " Copious water" Final Subject C. Indican (mg.) 25-3 0.729 21.3 0.544 21. 2 0.497 15-5 0.507 24.6 Bacterial nitrogen (gm.) 0.543 Subject V. Indican fme.) 72.2 0.899 79.2 0.822 77-4 0.875 64.9 0.675 92.7 Bacterial nitrogen (gm.) 0.693 The data in Table 3 indicate that copious water-drinking pro- duced a marked reduction in the processes of intestinal putrefaction, as measured by the urinary indican Output. The excretion of bac- terial nitrogen in the feces was practically at its minimum simulta- neously with the low indican values. In the period following the "high water" Ingestion, this uniformity of relationship is lost inas- much as the indican values undergo a sharp rise, whereas the bac- terial values are increased only slightly. 1913] N. R. Blatherwick and P. B. Hawk 39 It is evident that any uniform relationship between urinary in- dican and fecal-bacteria excretions, over any period of time and under various dietary conditions, is probably accidental. Indican, of course, has its origin in the indole which is produced f rom protein in the intestine through the activity of the indole-forming bacteria. If all the bacteria present in the intestine were indole- formers, then some definite relationship could reasonably be expected between the urinary indican and the fecal-bacteria nitrogen, provided that indican is a reliable putrefaction-index. However, inasmuch as there are several species of intestinal bacteria which are not classed as indole- organisms, it is readily seen how variations in the growth and development of these types of bacteria will influence the fecal- bacteria nitrogen values, but will have no influence upon the Output of urinary indican. Conclusions. When 500 c.c. of distilled water were added to the usual water-ingestion at each meal (100 c.c), a decrease was noted in the amount of bacterial nitrogen excreted daily in the feces. This held true for two subjects. One subject responded more freely to the influence of the water than did the other. When the water-ingestion (100 c.c.) was increased by 850 c.c. per meal, a more pronounced decrease in the daily excretion of bacterial nitro- gen was observed. This was more emphasized in the one case than in the other, but was very obvious in both. Since the amount of bacterial nitrogen occurring in the feces may, in a way, be considered an index of the utilization of the pro- tein in the food, we are led to conclude that there was a more effi- cient utilization of the proteins and hence better digestion and absorption when water was taken with meals. In both cases the beneficial results were not confined to theperiodsof increased water- intake, but continued into the periods following. Two subjects fed upon a uniform diet for a period of slightly more than one month were found to have an average content of 57.54 percent of bacterial nitrogen in the fecal nitrogen. The average amount of dry bacteria excreted per day was 6.189 gm. The Proportion of dry bacteria in dry feces was found to be 29.94 percent, A decreased Output of urinary indican was observed to accom- 40 Studies on Water Drinking [Od pany the copious water-ingestion. There was, however, no definite relationship betvveen the values for urinary indican and fecal- bacteria nitrogen under all conditioiis. A definite relationship would probably be accidental. BIBLIOGRAPHY 1. FowLER and Hawk: Jour. Exper. Med., 12, 388-410 (1910). 2. Mattill and Hawk: Jour. Amer. Chem. Soc, 33, 1999-2019 (1911). 2^ Bergeim and Hawk: Ibid., 35, 1049 (1913). 3. Findlay: Physical Chem. and its Applications in Medical and Biological Science, London, 1905. 4. HowE, Mattill and Hawk: Jour. Biol. Chem., 11, 103 (1912). 5. Koeppe: Deut. med. Woch., 624 (1898). 6. Nocht: Hyg. Rund., 273 (1892). 7. Winkler: Zeit, physikal. diät. Ther., 8, 671 (1905). 8. Harlow: Quoted by Oehler (10). 9. Spitta: Rubner's Handbuch Hygiene, Leipzig, 191 1, p. 28. 10. Oehler: Münch. med. Woch., 59, No. 50 (1912). 11. Mattill and Hawk: Jour. Exper. Med., 14, 433 (1911). 12. Harris: Jour. Am. Med. Assn., 59, 1344 (1912). 13. Ehrenpfordt: Zeit. f. exper. Path. u. Therap., 7, 455 (1909). 14. Strasburger : Zeit. f. klin. Med., 46, Nos. 5 and 6. 15. Sato: Zeit. f. exper. Path. u. Therap., 7, 427 (1909). 16. Berger and Tsuchiya: Zeit. f. exper. Path. u, Therap., 7, 439 (1909). 17. MacNeal, Latzer and Kerr: Jour. Infect. Dis., 6, 147 (1909). 18. Schittenhelm and Tollens: Centbit. f. Inn. Med., No. 30 (1904). 19. Lissauer: Arch. f. Hyg., 58 (1906). 20. Tobaya: Iji Shimbun (Med. Zeit), 758 (1908). 21. Sherwin and Hawk: Unpublished data. A NOTE ON THE DETERMINATION OF AMMONIA IN URINE STANLEY R. BENEDICT and EMIL OSTERBERG (Department of Chemistry, Cornell University Medical College, New York City) Since the proposal by Folin^ of his air-current method for the distillation of ammonia, this procedure has been almost universally adopted for estimating ammonia in urine, and in other fluids or mixtures where unstable nitrogenous substances may be present. The accuracyof this method, as employed under ordinary conditions, has never heretofore been questioned. Steel,^ however, in a de- tailed examination of the FoHn method under certain conditions, has called attention to the fact that the alkah employed by FoHn (sodium carbonate) is incapable of completely decomposing mag- nesium ammonium phosphate, and in case the urine contains appre- ciable quantities of this substance, results are low and very irregulär. As urines are not infrequently met with which contain magne- sium ammonium phosphate crystals, Steel proposed the modified technique of using one gram of sodium hydroxide and fifteen grams of sodium chloride, in place of the carbonate recommended by Folin. Steel shows conclusively that this mixture of sodium chloride and hydroxide will liberate ammonia quantitatively from magnesium ammonium phosphate, and details a number of experiments to show that substances other than ammonium salts which might be expected to occur in urine are not decomposed by this treatment so as to yield any ammonia. Steel also gives figures of numerous determinations made on urines which contained no preformed magnesium ammonium phos- phate crystals, to show that results are identical under such condi- tions, whether sodium carbonate or sodium hydroxide and sodium chloride be used to liberate the ammonia. 1 Folin: Zeit. f. physiol. Chetn., 37, p. 161 (1902). 2 Steel: Jour. of Biol. Chem., 8, p, 365 (1910). 41 42 Determination of Ammonia in Urine [Oct. During the past two years we have made it a practice to run two ammonia determinations on each sample of urine, in one of which sodium carbonate was employed to liberate the ammonia, while in the second one Steel's mixture of sodium hydroxide and sodium Chloride was used. Several hundred determinations hiave been made in this way, including both dog and human urines, and the figures thus obtained do not corroborate those reported by Steel for similar determinations. In every sample of urine analyzed we have obtained slightly higher figures where the hydroxide-chloride mixture was used, than where carbonate was employed. There would be little object in including here our large mass of figures obtained in this connection. We may summarize them by stating that normal and pathological urines were used, very few of which were not strongly acid in reaction, and none of which showed the presence of any magnesium ammonium phosphate crystals. Where Steel's hydroxide-chloride mixture was used results were always higher by o.i to 0.8 c.c. of n/io Solutions, amounting usu- ally to from one to seven percent of the total ammonia present, than where carbonate was used. The difference in the absolute amount of ammonia obtained by the two processes is obviously small, but the constant results raise a distinct question as to the essential accuracy of the two procedures employed. Does Folin's original process fail to yield all the ammonia from ordinary urines, or does Steel's modification decom- pose some additional urinary constituent in amount sufficient to yield distinctly measurable quantities of ammonia? This question puz- zled US for a long time, but we believe that we have found the cor- rect answer to it in a simple fact which has apparently been over- looked by both Steel and Folin in their work in this connection. If carbonate be added according to Folin's directions to a sample of urine, and the aeration process be carried out for a few minutes, and a few drops of the mixture be then examined under the micro- scope, ahiindant masses of magnesium ammonium phosphate crys- tals will he found. We have obtained this result with every sample of urine examined. It can be readily verified by shaking a sample of urine with a little sodium carbonate for about two minutes, and examining the mixture under the microscope. Whether air passes 1913] Stanley R. Benedict and Emil Osterberg 43 through the mixture is immaterial. As aeration proceeds, however, the phosphate is gradually decomposed, even by the carbonate, so that at the end of an hour the phosphate crystals may no longer be readily found. But once having had these crystals formed in con- siderable quantity we must conclude that the ammonia precipitated in this form is not quantitatively liberated again, because, as Steel has shown, starting with larger amounts o£ magnesium ammonium phosphate, one can scarcely recover fifty percent of the ammonia where carbonate is used. The abundant formation of the triple phosphate in the early stage of Folin's original method accounts fully for the slightly higher figures we have obtained where hydrox- ide was the alkali employed. It also accounts for the long time fre- quently necessary to obtain a maximal yield of ammonia in the Polin process. Using the same air-current, we have observed that either carbonate or hydroxide would give theoretical results at the end of an hour, using pure ammonium chloride Solution, whereas in the case of urine, sodium hydroxide would still give a maximal yield in one hour, while sodium carbonate required nearly two hours to give its maximal yield. In the light of the above-mentioned facts it is obvious that if Steel's Observation that magnesium ammonium phosphate is not quantitatively decomposed by carbonate be correct (and we have ourselves repeatedly verified this conclusion) , it f ollows as a matter of course that Steel's modification must give higher figures with all urines, and this is exactly what we have found to be the case. The difference appears to be in favor of Steel's modification. We cannot account for Steel's results in this connection. As a conclusion we wish to point out that the question of triple phosphate formation, and its incomplete decomposition by carbonate, is one which applies to all urines in varying degrees, and we are of the opinion that whatever procedure is used, one should be adopted which will ensure complete decomposition of the phosphate whether originally present in the urine or not. In discussing Steel's findings Folin^ recommended that urines which contained crystalline mag- nesium ammonium phosphate should first be treated with sufficient acid to dissolve the crystals, after which seven to ten grams of potas- 3Folin: Jour. Biol. Chem., 8, p. 497 (1910). . 44 Determination of Ammonia in Urine [Oct. sium Oxalate and one gram of carbonate are added to twenty-five c.c, and the aeration process carried out as usual. This procedure will prevent reformation of the triple phosphate. Folin preferred this procedure to that suggested by Steel because he believes that carbonate is a far safer alkali to employ than is hydroxide. If, however, as we have pointed out above, a method should be em- ployed which will prevent interference by magnesium ammonium phosphate in all urines, the addition of the large quantity of Oxalate (seven to ten grams) in every determination, is somewhat of a draw-back. We are inclined to regard Steel's modification as a safe procedure until someone shows that it actually decomposes other urinary constituents to yield ammonia. We feel that it is distinctly preferable to carbonate alone, as advised in the original Folin process. We may add that in his latest " microchemical " processes for ammonia, Folin and his collaborators^ have provided for the addi- tion of Oxalate in all instances. The small volumes dealt with keep down the quantity of Oxalate necessary. The figures reported by Folin and Macallum by the colorimetric method for ammonia in urine do not seem to us very satisfactory. While in many instances the agreement between the figures obtained by the old process and by the new is very close, in several cases the colorimetric values ex- ceed the others by from five to twelve percent of the total ammonia present. Folin and Macallum State that "a trace of something capable of giving a color with Nessler's Solutions continues to come long after all the ammonia has been removed," but that "the effect of this substance in actual ammonia determinations is so small as to be hardly, if at all, perceptible." So long as this substance capable of giving a color with Nessler's Solution remains an unknown factor and where results may be as much as twelve percent higher by the colorimetric procedure, we should hesitate to regard this latter proc- ess as of equal accuracy with the older. * Folin and Macallum: Jour. Biol. Chem., ii, p. 523 (1912). Folin and Denis: Ibid., II, p. 532 (1912). STUDIES OF AERATION METHODS FOR THE DE- TERMINATION OF AMMONIUM NITROGEN 3. The ammonium nitrogen in beef* JACOB SHULANSKY and WILLIAM J. GIES {Biochemical Laboratory of Columbia University, at the College of Physicians and Surgeons, New York) I. Introduction. A modification of the original Folin METHOD. Several years ago, during the course of a series of €x- periments on the effects of magnesium sulfate on metabolism, Steel^ obtained anomalous results with the Polin method for the determi- nation of urinary ammonium. The senior author, who was directing the experiments, suggested that these anomalous results might be due to the formation of ammonio-magnesium phosphate and to in- complete ejection, by the Folin method, of the ammonium from this Compound. With Steel,^ he then ascertained that the Folin method was inadequate for the determination of the yield of ammonia from urine that contained crystalline ammonio-magnesium phosphate. They found that sodium carbonate, in the proportions employed for the Folin method, was unable to disengage more than about 40 percent of the ammonium nitrogen from moderate quantities of ammonio-magnesium phosphate, whereas 0.5-1 gram of caustic alkali (instead of carbonate) promptly ejected all of it from large amounts of such phosphate. " Even in the presence of 50 times its weight of sodium carbonate and with 10 hours of thorough aeration, crystalline ammonio-magnesium phosphate cannot be completely decomposed by the Folin method of ammonia determination" (p. 81). Steel and Gies wrote as follows at the conclusion of their paper : " We hope in the near f uture to report a simple modification * The previous papers in this series were those by (i) Steel and Gies: Jour. Biol. Chem., 1908, v, p. 71, and (2) Steel: Ibid., 1910, viii, p. 365. 1 Steel : Ibid., 1908, v, p. 85. (Dissertation submitted in partial fulfilment of the requirements for the degree of Ph.D. at Columbia University.) 2 Steel and Gies: Ibid., 1908, v, p. 71. 45 46 Studies of Aeration Methods [Oct of the Polin method that will yield all the ammonia (nitrogen) from ammonio-magnesiiim phosphate in the urine without produc- ing ammonia from non-ammoniacal radicals" (p. 83). The modification Steel and Gies had in mind when this inten- tion was expressed, was, as their preliminary results had suggested, the Substitution of caustic alkali for sodium carbonate. For some time, thereafter, Steel continued the work along these lines, in this laboratory, and found^ that 0.5-1 gram of sodium hydroxid plus about 15 grams of sodium chlorid may be substituted for the sodium carbonate and sodium chlorid in the Folin method, for the com- plete ejection of ammonium from crystalline ammonio-magnesium phosphate in urine, without producing ammonia from amino radicals. This modified Folin method has been in regulär use in this laboratory ever since, for the determination of urinary ammonia, because it appears to be quite as efficient as the original Folin proc- ess in all respects and is certainly much more accurate when an appreciable quantity of ammonio-magnesium phosphate is present. Folin's criticism of the proposed modification. Shortly after the publication of this finding, Folin wrote, in part, as fol- lows:^ "The possible occurrence of minute amounts of ammonium magnesium phosphate hardly Warrants the Substitution of sodium hydrate and sodium chloride for sodium carbonate and sodium Chlo- ride in all ammonia determinations as recommended by Steel. There can scarcely be any doubt but ( !) that the carbonate is the safer reagent, and therefore to be preferred, unless weightier reasons can be found against it than the possible ( !) occurrence of traces of the triple phosphate." In expressing these opinions, Folin appears to have ignored the data which showed that ammonia was not evolved, by the NaOH- NaCl process, from such urinary constituents as allantoin, creatin, Creatinin, glycocol, guanin, hippuric acid, leucin, taurin, tyrosin, Urea, uric acid. The only support that Folin published for his opinion against the use of sodium hydroxid appears to be the fol- lowing (p. 497) : "The use of sodium hydrate in connection with the air current method for determining ammonia in urine was advo- 3 Steel : Jour. Biol. Cheni., 1910, viii, p. 365. * Folin: Jour. Biol. Chem., 1910, viii, p. 497. 1913] Jacob Shulansky and William J. Gies 47 cated by Moritz several years ago, but it appears to have failed to meet with much approval : Arch. f. klin. Med., Ixxxiii, p. 567, 1905." This quotation virtually begs the question. Results of experiments shozving " that the carbonate is the safer reagent," instead of com- ment on the failure of Moritz's Suggestion "to meet with much approval," would have been convincing. With all due respect for Folin's opinion, we must await a demonstration of the superiority of sodium carbonate over sodium hydroxid in the aeration process, before we can agree with Polin that the modified method is not de- cidedly better than the old. Application of the modified method to beef. In October 19 IG, soon after Dr. Steel's departure from this laboratory to ac- cept a professorshipat the University of Missouri, we began a series of experiments to determine the efficiency of the NaOH-NaCl method when applied to the determination of the ammonium nitro- gen in beef. It appeared probable that the protein in meat would yield, under such conditions, particularly large proportions of am- monia as a result of hydrolysis by the caustic alkali ; and we desired to ascertain the degree of deficiency, if any, of the method from this Standpoint. Most of the data recorded in this paper were obtained by the junior author in iqig-'ii, but verifications were occasionally con- ducted by the senior author during that year and the ensuing one. Publication of the results has been purposely delayed in order that the data might accompany the succeeding paper by Dr. Smith,^ whose work was begun in September, 191 1. 2. Comparative efficiency of the NaaCOg-NaCl and NaOH- NaCl aeration methods for the determination of ammonium( nitrogen. I. Fresh beef. In our direct comparison of the effi- ciency of the two aeration methods when applied to meat, we con- sidered it possible that (A) the protein of meat might yield consid- erable ammonia by hydrolysis, and (B) that some of this ammonia might combine in part with magnesium and phosphate in the meat to produce ammonio-magnesium Phosphate ;*5 (C) that the ammonium 5 Smith : Biochemical Bulletin, 1913, iii, p. 54. « Crystals of ammonio-magnesium phosphate form promptly in muscle fibers exposed to ammonia fumes or immersed in ammoniacal Solutions. 48 Studies of Aeraiion Methods [Oct in normal muscle plasma occurs, in part, in the form of ammonio- magnesium phosphate or may readily pass into that Compound ;'^ (D) that the protein of meat, by combining with the alkali, might reduce the power of the latter to eject the nitrogen from ammonio-mag- nesium phosphate in the aeration process; and (E) that sodium hydroxid and sodium carbonate would behave very differently in de- grees of hydrolytic influence on the protein or other unstable con- stituents, as well as in degrees of decomposing action on any am- monio-magnesium phosphate present to begin with or produced by the treatment. The work was planned under the influence of these possibilities. General method. Hashed fresh beef (15-25 gm.), after accu- rate weighing, was thoroughly triturated with sand and water in a mortar, and transferred to a tall cylinder of suitable width. The total amount of added water was 100 c.c. About 2-3 gm, of sodium hydroxid or 4 gm. of sodium carbonate were added, kero- sene promptly poured in, and strong aeration immediately hegiin and continued 8-12 hours. The force of the current was practically the same for each sample. Each alkali was used without addition of sodium chlorid, so that maximal effects, good and bad, might be elicited. The essential data are summarized in Table i. TABLE I Data showing the yields of ammonium nitrogen from hashed beef, in compara- tive aerations with sodium hydroxid and sodium carbonate t»« Date, 1911 Aeration with NaOH Aeration with NajCOs nee in favor he NaOH method 0^ Meat taken Ammonium N IC30 g^. me«t Meat taken Ammonium N icx> gm. meat I 2 I 2 Av. I 2 I 2 Av. rt - - Grams Milligrams !« Grams Milligrams ?< i A A A B C C Mar. 15 Mar. 16 Mar. 18 Mar. 25 May 3 May 5 26.896 24.230 19.146 19.596 25-797 16.085 27-379 27-257 17.470 16.526 27.204 15-969 36.44 28.89 21.21 50.368 28.437 30.463 27.61 29-79 28.05 50-998 35-612 31-561 0.032 0.029 0.025 0.051 0.032 0.031 21.727 27.261 20.932 27.220 22.424 26.552 21.912 15-878 24.694 22.234 16.75 1387 16.72 26.847 25.410 13-71 15-33 41.79 21.03 28.90 0.017 0.014 0.016 0.042 0.024 0.027 o.ois o.ois 0.009 0.009 0.008 0.004 ^ In the putrefaction of meat, crystals of ammonio-magnesium phosphate appear early and accumulate rapidly. * Each reserve supply of hashed beef was kept frozen during the experiment. 1913] Jacoh Shulansky and William J. Gies 49 In repetitions of the tests, with sodium carbonate and sodium chlorid in accord with Folin's original directions, and with sodium hydroxid and sodium chlorid plus moderate quantities of alcohol to reduce frothing, the senior author has obtained somewhat lower values for each method. Our results in the latter connection will be published in a discussion of another phase of the work. It is obvious, from the data in Table i, that the sodium hydroxid treatment yields somewhat more ammonia than the treatment with sodium carbonate. Whether some or all of the surplus was due to ammonia that had been produced from other sources than ammo- nium radicals will be discussed in a future consideration of related data. It may be said, in anticipation, however, that such surpluses appear to be derived wholly from ammonium radicals, especially when a moderate quantity of sodium chlorid is present and the periods of aeration are not excessive — ^they were intended to be in these tests. The sodium hydroxid data are somewhat higher than those already reported, for "fresh" meats, as obtained by methods that do not yield the füll proportion of ammonia from triple phos- phate (see page 65).^ II. Ammonio-magnesium PHOSPHATE. In Table 2 we give re- sults, obtained independenfly by the junior author, Avhich confirm the earlier observations by Steel and Gies. III. "MoDiFED"''' BEEF. A. Hashed heef kept at refrigerafion temperatures. " Fresh " beef was prepared and ref rigerated by our own methods, as for use in nutrition experiments.^ Five prepara- tions from as many independent supplies of fresh beef were made successively on Oct. 29, Oct. 31, Nov. i, Nov. 3, and Nov. 5, 1910. Table 3 gives the analytic data obtained by the junior author with the NaOH-NaCl method. Occasional comparative determinations with both methods by the senior author not only checked, in a general way, the data in Table 3, but also indicated that the higher the yield of ammonia with the NaOH-NaCl method, the lower the comparative yield with 8 By "fresh" meat we mean, in the description of our general method (p. 48), beef that was purchased as such. The periods and conditions of previous refrigeration were unknown to us. 8 Gies : Amer. Jour. PhysioL, 1901, v, p. 235 : Biochemical Researches, 1903, i, repr. i ; Proc. Soc. Exp. Bio!. Med., 1908, vi, p. 27. 50 Studies of Aeration Mcthods [Oct. TABLE 2 Data pertaining to the comparative yields of ammonium nitrogen, by ihe NaOH-NaCl and NdCOs-NaCl aeration methods, from crystalline ammonio-magnesium Phosphate* Aeration with NaOH Aeration with NajCOg Phosphate taken Total nitrogen Phosphate taken Total nitrogen Found Per gram Found Per gram Gram Milligrams Gram Milligrams 0.2695 0.3676 0.4383 0.5053 0.4718 0.4447 22.39 29-94 36-93 42.63 40.82 ■iO.lA 83.09 81.45 86.54 84.38 86.51 88.02 0.2724 0.1564 0.3109 0.3968 0.3394 7.14 3-92 8.269 8.689 8.409 26.21 25.06 26.60 21.81 24.78 Average 84.99 Average 24.89 Average loss, per gram of phosphate, by the Na2C03 method 6o.iot the NaaCOg-NaCl process — the greater the proportion o£ ammonio- magnesium phosphate in the refrigerated meat, the smaller the pro- portion of ammonia ejected by the NaaCOg-NaCl method. The data for the latter method are accordingly omitted from the tables. TABLE 3 Data pertaining to the yield of ammonium nitrogen, by the NaOH-NaCl method, from five preparations of hashed fresh heef, at intervals during prolonged periods of storage at different refrigeration temperatures Preparation I. A. Oct. 29, 1910-Jan. 5, 191 1 at 26° F. Meat taken, grams Ammonium nitrogen, milligrams Date I 2 Average I 2 Average Per Cent. 10-29, '10. . . 33-730 27.820 30-775 3-54 3.26 3-40 O.OII 11-5, *I0... 25-393 28.377 26.885 4.96 3-54 4-25 0.016 11-15, '10.. . 25-635 25-258 25-447 3-97 4-39 4.18 0.016 12-1, '10... 19-125 19.025 19-075 4-25 3.83 4.04 0.021 1-5. 'II--- 18. 119 23.942 21.031 6.66 6-94 6.80 0.032 B. Jan. S-Feb. 13, 191 1 in ordinary refrigeratorj 1-23. II. 2-13. 'II. 31-973 23-647 33-185 22.398 32.579 23-023 57.39 75-70 57-25 70.85 57-32 73-28 0.174 0-313 * Data pertaining to total nitrogen in the phosphate, as determined by the Kjeldahl method (0.3883, 0.2439 and 0.3465 gm. of phosphate taken) : total nitrogen per gram of phosphate— 87.34, 89.64 and 88.10 (av. 88.36) mg. t A loss of 70.7 per cent. t The temperature in the ordinary house refrigerator varied between 47-58° F. I9I3] Jacob Shulansky and William J. Gies 51 TABLE 3 (continued) Preparation II. A. Oct. 31, 1910-Jan. 9, 191 1 at 26° F. Meat taken, grams Ammonium nitrogen, milligrams Dale I 2 Average I 2 Average Per Cent. II-I, '10... II-II, '10... 11-16, '10. . . 12-2, '10... 1-9, '11... 20.665 20.059 23-580 21.447 20.276 32.607 20.042 29.109 26.283 23-522 26.636 20.051 26.345 23-863 21.899 4.26 4-25 10.36 4.67 6.43 9.08 3-83 4.86 6.52 6.16 6.67 4-04 7.61 5-59 6.29 0.024 0.020 0.028 0.023 0.029 B. Jan. 9-Feb. 14, 191 1 in ordinary refrigerator§ 1-23, II. 2-14, '11. 33-195 27.245 32.280 32.908 32-738 30.078 58.02 65.24 53-12 128.85 55-57 97-05 0.170 0-315 Preparation III. A. Nov. i, 1910-Jan. 12, 1911 at 26° F. II-I, •10... 30.158 35-551 32.855 12.90 8.65 10.77 0.034 II-II, '10... 19.254 25.082 22.168 5-22 5-39 5-30 0.024 II-2I, '10. . . 27.610 28.740 28.175 6-23 6.80 6.52 0.023 I2-S, '10. . . 17-832 21.147 19.490 4.67 5-39 5-03 0.026 I-I2, 'II... 20.716 22.932 21.824 10.07 8.93 9-50 0.044 B. Jan. I2-Feb. 15, 191 1 in ordinary refrigerator 1-27, II. 2-15, 'II. 35.051 28.932 24.491 27.583 31.982 26.037 50.29 87.36 77-44 85-82 63-87 86.59 0.200 0.333 Preparation IV. A. Nov. 3, 1910-Jan. 13, 191 1 at 26° F. II-3. 'lO... 27-485 36.518 32.002 4-39 5-52 4-96 0.016 II-I4, '10... 19-523 24.396 21.960 2.98 9-77 6.38 0.028 11-22, 'lO. . . 20.725 22.260 21.493 5-53 3-97 4-75 0.022 12-6, '10... 22.824 27-757 25.296 4-39 4.82 4.61 0.018 I-I3. 'II... 28.519 J 41-341 34-930 10.48 12.86 11.67 0.034 B. Jan. 13-Feb. 20, 1911 in ordinary refrigerator 1-27, II.. 2-20, '11 . . 28.059 31-995 28.155 32.596 28.107 32.296 55-78 96.74 45-01 102.20 50.40 99-47 0.179 0.308 Preparation V. A. Nov. 5, 1910-Jan. 17, 191 1 at 26° F. ii-S, '10... 28.163 25-348 26.756 3-82 5-52 4.67 0.018 11-14, '10. . . 18.544 36.604 27-574 1.84 5-38 3-62 0.012 11-28, 'lO... 20.423 19-965 20.194 3-12 2.98 3-05 o.ois , 12-8, 'lO... 23.780 21. 211 22.496 5-38 3-97 4-67 0.021 I-I7, '11. . . 31.804 24.966 28.385 8.81 6-57 7-69 0.027 B. Jan. 17-Feb. 25, 191 1 in ordinary refrigerator. 1-24, II. . . 2-25, '11. -■ 21.247 22.062 19-134 25-177 20.191 23.620 23-63 82.75 17-89 112.42 20.76 97-58 0.102 0.4II § The temperature in the ordinary house refrigerator varied between 47-58° F. 52 Studies of Aeration Methods [Oct. B. Hashed heef with definite additions of ammonio-magnesium Phosphate of known nitrogen content. The meat preparations re- ferred to in Table 3 acquired a content of er y stalline ammonio- magnesium phosphate after their storage in an ordinary refrigerator. We attributed the increased yield of ammonia nitrogen, as the examinations progressed (Table 3), to the accumulation of am- monium in the form of triple phosphate and other Compounds, as a result of bacterial activity.^^ The capacity of the NaOH-NaCl method to eject all the ammo- nium nitrogen from crystalline ammonio-magnesium phosphate in meat was tested directly by adding weighed amounts of such phos- phate, with known nitrogen content, to meat whose yield of am- monia had been determined, in other portions, before the triple phosphate was admixed. The analytic data are presented in Table 4. TABLE 4 Data pertaining to the yield of ammonium nitrogen, by the NaOH-NaCl method, from meat and triple phosphate mixtures of known independent yields of ammonium nitrogen by the same method Date Meat taken A Phosphate taken Total yield of ammonium nitrogen Prepa- ration* B A+B Grams Gram Found, mg. Calculated, mg.f Loss, mg. I II 311 JV y 12-9, '10 1-5. 'II 1-9. '11 1-12, '11 1-12, '11 1-13. 'II 12-12, '10 12-12, '10 1-17, '11 1-17. '11 26.886 32.513 25-942 28.703 24.428 24.018 25-957 24.489 28.243 29.584 0.2398 0.3763 0.3081 0.3514 0.4566 0.5781 0.2954 0.2734 0.3604 0.3393 18.4 39-3 31-3 35-1 47-2 48.9 28.2 21.8 35-3 31-7 26.0 42.4 33-6 42.3 49-5 57-2 30-4 28.2 38-2 36.5 7.6 3-1 2.3 7.2 2-3 8.3 2.2 6.4 2.9 4.8 10 The meat was ordinary hashed beef, which had been thoroughly mixed by hand before refrigeration. No attempt was made to prevent introduction of bacteria. * The preparations referred to in Table 3 were used. t The calculated total was derived, for the meat, from the data f or the nearest date (Table 3) ; for the phosphate, from the aeration data in Table 2. Thus, for the first total (26.0 mg.) we calculated: (a) 26.886X0.21 = 5.646; (fe) 0.2398X85 = 20.383; (c) 5.646 + 20.383 = 26.029. It is obvious that such calculations are at best close approximations and that the average "loss" may be due, in part, to overcalculation. 1913] Jacoh Shulansky and William J. des 53 The data in Table 4 suggest that the protein of the meat inter- fered with complete liberation of the ammonium nitrogen from the very large proportions of triple phosphate present in each case, but the probabihty of overcalculation of the amounts of ammonium ni- trogen in the mixtures is against that indication. (See footnote. Table 4. ) The data in Table 4 also suggest that ammonia was not produced from the protein of the meat. 3. General conclusions. Ignoring, for the present, certain considerations which the senior author intends to discuss in a sub- sequent issue of the Biochemical Bulletin^ we conclude that the NaOH-NaCl method for the determination of ammonium nitrogen in meat is more accurate than the NagCOs-NaCl process. This greater degree of accuracy we attribute to the capacity of the NaOH-NaCl method to eject, as ammonia, all the nitrogen in ammonio-magnesium phosphate, whether crystallized or dissolved. It is evident, also, that the NaOH-NaCl method is particularly suitable for the study of meat subjected continuously to prolonged periods of cold storage, because of the completeness with which the nitrogen of ammonio-magnesium phosphate may be removed and ob- tained as ammonia, and because any deficiencies of the method, espe- cially as compared with the NasCOg-NaCl process, induce plus errors — errors of the kind that would tend to suggest putrefactive changes very early, if any such changes were in progress. The ammonia determinations in the work on fish described by Smith^^ and by Perlzweig and Gies,^^ in the two succeeding papers, were based on these findings. Their results with the NaOH-NaCl method give special emphasis to the foregoing conclusions. [The preceding paper by Benedict,^^ which was received on the day the manuscript of this number of the Bulletin was forwarded to the printer, also has an important bearing on these deduc- tions. Ed.'\ 11 Smith : Biochemical Bulletin, 1913, iii, p. 54. 12 Perlzweig and Gies : Ibid., p. 69. 13 Benedict and Osterberg: Ibid., p. 41. A STUDY OF THE INFLUENCE OF COLD-STORAGE TEMPERATURES UPON THE CHEMICAL COMPOSITION AND NUTRITIVE VALUE OF FISH CLAYTON S. SMITH (Biochemical Laboratory of Columbia University, at the College of Physicians and Surgeons, New York) I. INTRODUCTION With the invention of the ammonia machine for the production of low temperatures, the cold-storage industry may properly date its beginning. Although the use of low temperatures for the preservation of foodstuffs has long been recognized, it is only recently that careful studies have been made of the effects upon food products of long periods of cold storage. Among the first to note the effect of cold upon foods was Tel- lier/ who observed that meat stored at from — 2° to +3° C. retained its freshness. Bouley^ also found that at a temperature of — 2° to +3° C, meat kept for an indefinite period so far as putres- cence was concerned, but not from the Standpoint of edibility. Grassman^ found that meats kept for eight months at a temperature of — 2° C. to — 4° C. did not deteriorate. He claimed that refrig- erated meats may be cooked in less time than the fresh materials. A comparative study of the chemical composition of fresh and cold-stored foods was made by Girard,"* who studied the phos- phorus content of vegetables and some animal products, such as pork, mutton, beef, eggs and milk. Gautier^ made a very detailed study of the difference between fresh and cold-stored beef and iTellier: Rev. d. Hyg., 1897, xix, p. 298. 2 Bouley : Compt. rend., 1874, Ixxix, p. 739. 3 Grassman : Landw. Jahrb., 1892, xxi, p. 467, * Girard : Compt. rend., 1896, cxxii, p. 1387. 5 Gautier : Rev. d. Hyg., 1897, xix, p. 289. 54 1913] Clayton S. Smith 55 mutton. He found i per cent. less moisture in the cold-stored products than in the f resh, but no difference in digestibility. The effect of cold-storage upon bacteria and enzymes in meat was studied by both Mai^ and Müller,'^ who showed that putrefac- tion was prevented but that the action of the enzymes was not in- hibited. In the more recent investigations of the effects of cold- storage, emphasis has been placed upon its influenae on meat, poultry and game, and to some extent upon vegetables. Little, however, has been said concerning cold-stored fish. The gap in our knowledge at this point led to this investigation. In 1861, Enoch Piper^ established a plant in Beekman Street, New York City, for the freezing of fish by means of ice and salt. Davis,^ in 1868, invented special pans for the freezing of fish, but used the same refrigerating agent. The first carload of frozen fish was shipped from Oregon to New York in 1883, but the quantity was very small compared with the large shipments today. As early as 1888, Russia had an important frozen-fish industry. Sturgeons and dolphins were the principal fish used and the freezing was generally conducted in cellars at the sea-shore. The frozen-fish industry began in America in the early 90's. The first plant was established at Sandusky,^*^ Ohio, in 1892. The industry progressed slowly at first because of a strong prejudice against cold-stored products and particularly against frozen fish. Salmon was at first practically the only fish frozen, but at the pres- ent time many varieties are refrigerated. Each refrigerating concern may have its own particular method of freezing fish but the general practice seems to be to freeze the fish, dip them in water, and re freeze in order that they may be com- pletely encased in ice. They are then stored at a temperature of — 16° C. The coating of ice prevents loss of water due to surface evaporation. This coating is renewed as occasion requires. Much has appeared in the literature concerning processes for the production of low temperatures and methods for handling cold- ß Mai : Zeit. Nähr. u. Genus., 1901, iv, p. 18. ^Müller: Arch. f. Hyg., 1903, xlvii, p. 127. 8 See Loverdo : Le Froid Artificiel, 1903, p. 401. ö Davis : Ice and Refrigeration, 1901, xxi, p. 93. 10 Ibid. 56 Cold-Storage of Fish [Oct. stored products. Little, however, has been written concerning the effect of cold-storage upon the chemical composition of the flesh of fish. The Report of the U. S. Commission of Fish and Fisheries, for 1888, contains data of analyses of American food fishes. The specimens were, for the most part, fresh fish, a few being pre- served but none were cold-stored. More recent analyses of fresh fish have been made by WiUiams and by Ulrich.^ ^ WilHams'^^ work was conducted from an economic Standpoint, while that of Ulrich was a purely chemical study of the composition of fish flesh. It happens that both of these authors have analyzed specimens of fish belonging to species similar to those analyzed by us. Mention will be made of their results when our own are discussed. The work here reported was undertaken with the hope of ascer- taining what change, if any, fish muscle undergoes during long periods of cold-storage. In order that our experiments might be properly controlled, a preliminary study was made of the muscle of fresh fish. II. EXPERIMENTAL Preliminary handling of the fish. The fish used were the fluke, also known as summer flounder {Paralithys dentatus Linn.), and the winter flounder {Pseudoplenronectes americanus Walb.), both of which were furnished by a reliable dealer. These fish were selected because their habits imply that they might be particularly prone to bacterial decomposition in cold-storage. The flounder is peculiarly a "bottom fish," in fact is in the mud or sand most of the time. The various lots of fish were taken from the dealer's ordinary commercial products, which had been handled from the water to Fulton Market (N. Y.), and in the cold-storage plant, in accordance with the practical methods of the trade. As soon as a catch arrived at the wharf, three fish were sent to the laboratory and twenty-four others put into a cold-storage plant. At the plant the fish were suitably dipped, f rozen and cold-stored as usual. Those which came to the laboratory arrived packed with cracked ice in an 11 Ulrich: Arch. Pharm., 191 1, ccxlix, p. 68. 12 Williams: Chetn. News, 191 1, civ, p. 273. 1913] Clayton S. Smith 57 ordinary willow basket. The basket was lined with a water-proof paper, covered with burlap and the whole wrapped in heavy manila paper. The time in transit from the wharf to the laboratory was a little over an hour. Very Httle of the ice melted en route and, with one exception, the fish were never in contact with free water in the basket. Upon requisition, fish were taken from storage, wrapped sepa- rately in paper, and sent at once to the laboratory, where they arrived in a short time. No appreciable thawing took place in transit. The storage samples, if received during the winter months, were kept over night under paper Covers in shallow pans at room temperature. For the summer months this method of thawing was modified by placing the fish in an ordinary refrigerator over night and then allowing them to remain for an hour at room temperature the next morning. In either case, as soon as the fish were com- pletely thawed, the analyses were begun. Each lot of fish furnished by the dealer was given a number by him. The tag also bore the date and, in the case of stored fish, both the date of receipt at, and the date of delivery from, the cold- storage plant. In every instance before the fish was prepared for analysis, the general external appearance was observed and, upon dissection, the color and texture of the muscle were also noted. Fresh fish which have been out of the water for some time often show "clots" of slime over their bodies, and in their mouths and gills. Under such conditions the gills may be pale and the fins slightly reddened. When a frozen fish is allowed to thaw at room temperature its skin becomes distinctly dry after the water has evaporated, no slime ever forming. Its gills may be pale but the fins are not red. A slight yellowish brown discoloration, just under the skin, was noted in the case of stored fish ; in fresh fish there were certain gray subcu- taneous areas. A difference in the consistency of the muscle of the stored fish was also noticed, that of the fresh fish being the firmer. These observations apply only to fresh fish kept on ice from 48-72 hours and to cold-stored fish immediately after thawing. Analytic determinations and methods. The analytic determina- tions may be conveniently considered under four headings : First — 58 Cold-Storage of Fish [Oct. water, total solids, organic matter, inorganic matter ; second — ammonium nitrogen, total nitrogen, " soluble nitrogen," " insoluble nitrogen," " co- agulable nitrogen," " non-coagulable nitrogen;" third — lipins (per cent and acidity) ; fourth — reducing substances and acidity of aqueous extract. 1. The determination of ivater was made in the following manner: An accurately weighed sample, approximately 5 gm., was dried in a large porcelain crucible on a water bath for from three to four hours, after which it was placed in an air bath, at 110° C, and dried to con- stant weight. In the determination of ash, the following precaution was taken to prevent volatilization of Chlorides. After the organic matter had been completely charred over a low flame, an aqueous extract was made of the residue. This liquid was brought to the boiling point and allowed to cool somewhat, when it was decanted as caref ully as possible through an ashless filter, most of the residue remaining in the crucible. After drying, the residue and filter were ignited over a bunsen burner to a white ash. The extract was then added to the residue in the crucible and evaporated to dryness on a water bath. Finally the crucible was heated over a low flame to remove residual carbonaceous matter. By this method the soluble salts were not subjected to the high heat of ignition. Both total solids and organic matter were calculated from data obtained by the foregoing methods. 2. Ammonium nitrogen was determined by the following modifica- tion of Folin's method.^^ Fifty grams of fish muscle were ground in a mortar with sand. The finely ground meat was suspended in a mix- ture of equal volumes of 95 per cent alcohol and water in an aeration cylinder. The mixture of alcohol and water was used in order to prevent excessive frothing during aeration. The volume in each case was approximately 200 c.c. Fifty gm. of pure sodium chloride and 4 gm. of pure sodium hydroxide were added to each 200 c.c. of Suspen- sion. As soon as the alkali was added, very vigorous aeration was begun and continued for at least four hours. Total nitrogen was determined by the Kjeldahl process. Oxidation was facilitated by the addition of a small piece of crystallin copper Sul- fate to the sulfuric acid. 13 Steel and Gies : Journal of Biological Chemistry, 1908, v, p. 71 ; Steel : Ibid., 1910, viii, p. 365 ; Shulansky and Gies : Biochemical Bulletin, 1913, iii, p. 45. 1913] Clayton S. Smith 59 " Soluhle nitrogen" was determined by the Kjeldahl method, in 10 c.c. of an aqueous extract prepared in the foUowing manner : Twenty gm. of water were added to each gm. of fish taken, but in prepar- ing the extracts, an allowance was made for the water content of the flesh, which was found to average 78.48 per cent. Approximately 50 gm. of flesh were used. After the extract-mixtures were prepared, they were shaken thirty times and allowed to stand over night. Then they were again shaken, allowed to settle and filtered. During filtra- tion care was taken to prevent losses by evaporation. " Non-coagidahle nitrogen" was determined directly as follows: 100 c.c. of extract, prepared by the foregoing method, were heated gently to boiling and then treated with 2 c.c. of a 2 per cent Solution of acetic acid. When the liquid had again been carried to the boiling point, the Solution was filtered directly into a Kjeldahl flask, Into the beaker in which the precipitation had been made, were poured 50 c.c. of distilled water. This was brought to the boiling temperature and then used at once to wash the precipitate on the filter. The precipitate was washed twice in this manner, after which the total ("non-coagu- lable") nitrogen in the combined filtrate and washings was determined as usual. From data obtained by the foregoing methods, the " insoluhle ni- trogen" and " coagxtlahle nitrogen" were determined by difference. 3. To prepare flesh for the determination of its content of lipins, muscle was quickly removed from the fish, passed through a hashing machine, and the hash dried at room temperature before an electric fan, after which the residue was pulverized in a drug mill. Mixtures of aliquot portions of each powder were used in the extractions, which were made by the Soxhlet method upon 60 gm. charges, whenever practicable. The acidity of the lipin mixture was determined by shaking the sample with 50 c.c. of 95 per cent alcohol to which i c.c, of i per cent Phenolphthalein Solution in 95 per cent alcohol had been added and titrating with n/$ alkali Solution. A blank was always run simulta- neously on the alcohol. 4. The reducing power of aqueous extracts of fish muscle, after removal of the protein, was determined by Benedict's^* method. The extract was made by treating each gram of fish with 4 gm. of water. Coagulable protein was removed by the method described above. We always ascertained the degree of acidity of the aqueous extracts, as prepared for the various nitrogen determinations. Fifth normal 1* Benedict: Joitrn. Am. Med. Assn., 1911, Ivii, p. 1193. 6o Cold-Storage of Fish [Oct. sodium hydroxide Solution was titrated against loo c.c. of the extract, using Phenolphthalein as the indicator. Conduct of the examinations. Our examinations of the fish were conveniently divided into three series: (I) On fresh fish, (II) on fish TABLE I Series I. Fresh flounders. Percentage data, except as noted Water Total solids Or- ganic mat- ter Ash Am- monium N Fish No. Nitrogen Reac- Fish No. Sol- uble Insol- uble Coagru- lable Non-co- agulable Total tion of aqueous extract * Ai 78.35 78.80 79.01 78.74 78.62 79.10 76.81 79-52 79-76 77-88 75-78 80.40 81-03 82.49 81.56 81.51 80.13 81.66 82.49 82.22 81.32 82.78 80.32 80.87 81.46 82.14 83.00 82.42 81.27 82.91 21.65 21.20 20.99 21.26 22.38 20.90 23.19 20.48 20.24 22.12 24.22 19.60 20.36 19.97 19-77 19.97 21.09 19.62 21.96 19.23 19.04 20.90 22.66 17-37 1.29 1.23 1.22 1.29 1.29 1.28 1.23 1.25 1.20 1.32 1.56 1.23 0.014 0.014 0.012 0.025 0.022 3.26 3.21 A2 A3 B4 BS B6 3.27 3.51 3-32 3-58 3-21 3-21 3-49 4.04 3-70 3-44 3.80 3.26 3.42 3-47 3.24 2.87 2.76 2.52 2.72 3.03 2.90 2.69 2.81 2.88 2.85 2.99 317 3-11 3.08 3-10 C7 C8 O.OII 0.026 0.023 0.025 0.029 Cg Dio Du Dl2 1-34 L3S L36 E13 i.ana 2.361 2.716 2.336 2.409 2. 511 2.282 1.752 1.695 1.774 I.921 2.018 1.727 1.838 1.958 2. 121 1.998 2.244 2.238 2.025 1.562 2.275 0.705 O.711 0.552 0.612 0.501 0.562 0.772 0.724 0.429 0.485 0.629 0.737 0.426 0.453 0.351 0.447 0.363 0.554 0.374 0.373 0.372 0.399 0.458 0.396 0.346 0.341 0.317 0.314 0.383 0.436 0.426 0-399 0.408 0.405 0-383 0-378 1.7; 00 P46 00 P47 126 0.852 127 0.759 J28;0.852 J29 ^0.746 130:0.932 K31 I0.985 E:32|i.si8 K33 0.825 20 P48 TO R49 3S R50 IG Rsi !■> 28 I-I37 0.445 0.381 0.380 63 48 Average 80.15 21.52 20.16 1.2S 0.0195 0.964 2.084 0.579 0.383 3.18 I 37 Lipins — Fish L34-R51, in three groups; aliquot portions of the fish in each groups were extracted together : Total amount of lipins in the fresh flesh (per cent) — 0.386, 0.360, 0.392; average, 0.379. Acidity (mg. of KOH to neutralize i gm. of lipins)— 131, I47, 132; average, 136. * Expressed as c.c. of n/5 sodium hydroxid Solution required to neutralize 100 c.c. of extract. 1913] Clayton S. Smith 61 stored for six months, and (III) on fish stored for nine months. The analytic determinations upon each series of fish were of the same kind, but for convenience they may be considered in three different groups. TABLE 2 Series II. Flounders after six months of cold storage. Percentage data, except as noted Water Total solids Or- ganic mat- ter Ash Am- monium N Fish No. Nitrogen Reac- Fish No. Sol- üble Insol- uble Coagu- lable Non-co- agulable Total tion of aqueous extract* A70 A71 A73 B74 B75 B76 C77 C78 C79 D80 78.00 78.28 78.02 77.06 77.82 77.80 79-57 79.72 78.62 79.29 80.85 79-33 22.00 21.72 21.98 22.94 22.18 22.20 20.43 20.28 21.38 20.71 19-15 20.67 20.70 19.47 20.73 21.59 20.82 20.93 19.17 1.30 1.25 1.25 1-35 1-36 1.27 1.26 0.006 O.OII 0.009 0.034 0.022 O.OIO O.OII 0.016 0.008 0.015 0.008 0.008 0.022 0.017 0.015 3.57 3-31 3-47 3.53 3-43 3.34 3.19 3.21 3-41 3-23 3.09 3-38 20.12 19.41 17.92 19-39 1.26 1.30 1-23 1.28 D81 D82 D83 D84 1 D85 L108 80.39 81.62 81.02 81.81 82.02 81.64 82.5s 80.30 81.04 82.58 79.78 82.41 82.40 82.54 82.49 80.54 80.53 E86 E87 £88 F99 Figo Fioi G102 G103 G104 Hios H106 H107 I-I4S 0.799 2.395 2.:;8t 0.714 0.430 0.572 0.586 0.597 0.545 0.437 0.483 0.431 0.469 0.373 0.373 0.362 0.362 0.309 0.341 3.54 3.38 3.43 3.06 3-21 3.22 2.99 2.84 2.95 2.79 2.54 2.61 !.'?•? L109 1-43 Liio 0.9451 2.485 0.959J 2.101 0.959 2.251 0.907 2.313 0.746 2.2AA 1.38 Mm I.2S M112 I.IO M113 1.20 N114 1.30 N115 0.825 2.015 I.IO N116 I.IO O117 0.864 I.Q26 0.566 0.474 0.588 0.298 0.296 0.245 I.OO O118 0.770 T.870 I.OO O119 0.833 1.777 0.90 P120 P121 P122 R123 R124 Average 80.13 21.30 20.01 1.29 0.0142 O.887J 2.179 0.545 0.351 3.19 I.17 Lipins — Fish L108-R124, in three groups; aliquot portions of the fish in each group were extracted together : Total amount of lipins in the undried flesh (per cent) — 0.362, 0.458, 0.277; average, 0.366. Acidity (mg. of KOH to neutralize i gm. of lipins) — 150, 138, 120; cverage, 136. * Expressed as c.c. of n/5 sodium hydroxid Solution required to neutralize 100 c.c. of extract. 62 Cold-Storage of Fish [Oct, The first groiip of determinations included water, ash, ammonium nitrogen and total nitrogen. The determinations were made upon fish from four of the dealer's lots in each of the three series of observations. The second groiip of determinations, comprising total, soluble and non-coagulable nitrogen, together with the reaction of the aqueous ex- tract, was made upon fish from seven of the dealer's lots (Series I). TABLE 3 Series III. Flounders after nine months of cold storage. data, except as noted Percentage Water Tota solids Or- ganic mat- ter Ash Am- monium N Fish No. Nitrogen Reac- Fish No. Sol- uble Insol- uble Coagu- Non-co- lable agulable Total tion of aqueous extract* A125 A126 78.06 77-97 78.55 79-48 77-82 79.14 78.70 77-65 79-37 76-74 79-00 78-97 82.45 84.24 80.55 73-98 80.87 81.49 83.02 81.76 81.68 81.82 81.69 82.38 81.90 82.18 82.51 21.99 22.03 21.45 20.52 22.18 20.86 21.30 22.35 20.63 23.26 21.00 21.03 20.59 20.76 20.31 19-34 20.89 19.67 20.05 21.12 X.4O 1.27 I.I4 I.18 1.29 I.I9 1.25 1.23 0.027 0.026 0.013 0.022 O.OIO 0.019 0.024 0.032 0.032 0.017 0.021 0.012 3.39 3-44 3.29 3-34 3-55 3-31 3.28 3.48 3.54 3-79 3-35 3-33 3-12 3-61 3-55 3-08 3-33 2.99 2.98 2.99 2.23 2.87 2-93 A127 B128 B129 B130 C131 C132 C133 D134 D135 D136 L149 L150 Lisi M152 M153 M154 N155 N156 NiS7 21-73 20.73 19.72 1-53 1.27 1.31 E137 E138 E139 F140 F141 F142 G143 G144 G145 H146 H147 H148 1.074 I.127 1.043 0.820 0.913 0.966 2.046 2.483 2.507 2.260 2-364 0.741 0.719 0.592 0.466 0.542 0.603 0.333 0.408 0.451 0.354 0-371 0.363 1.87 1.85 1.63 0.0"? 1.35 I.IO o.go 0.859 0.895 0.966 0.806 0.859 2. 121 2.095 1.264 2.064 2.071 0.548 0.519 0-655 O.511 0.548 O.311 0.376 O.311 0.295 O.31I 0.90 0.97 O158 O159 O160 0.97 0.88 1.20 P161 P162 P163 Average 80.24 21.55 20.45 1.28 0.0213 0-937 2.128 0.586 1 0.353 3." 1.23 Lipins — Fish L149-P163, in four groups; aliquot portions of the fish in each group were extracted together : Total amount of lipins in the undried flesh (per cent) — 0.442, 0.497, O.391, 0.442; average, 0.443. Acidity (mg. of KOH to neutralize l gm. of lipins) — 11 1, 130, I39. 130; average, 127. * Expressed as c.c. of n/5 sodium hydroxid Solution required to neutralize 100 c.c. of extract. 1913] Clayton S. Smith 63 When this group of determinations was repeated on Series II and III, four dealer's lots were used for each series. The third group consisted of determinations of lipins, water and reducing power. Because of the small percentage of fat in the fish, all of the dealer's lots (six) in this group were drawn upon in each of the three series of observations. In performing a particular set of analyses, samples of flesh were uniformly taken, as nearly as possible, from the same region. The selected parts were in the right angles made by a line running from the head to the tail and a line perpendicular to it about midway between the mouth and the tail. The skin on the back was opened, turned over, and flesh taken from all depths to the bone. Flesh near the viscera, as well as remote from them, were included. The skin was not analyzed. For convenience in Consulting the data, the following plan of tabu- lation was adopted. Each capital letter in the accompanying tables represents a dealer's lot of fish. The numeral following the capital letter indicates an individual fish. Thus, B5 (Table i) represents a particular fish that was fresh when analyzed. B75 (Table 2) indicates a particular fish of the same lot which had been placed in cold storage and analyzed six months later. The capital letter also indicates, in terms of the appended summary, the date (in 191 1) when a particular fresh fish was subjected to analysis (or placed in cold storage) : A-Sept. 9- E-Oct. 18. I-Nov. 27. M-Dec. 6, B-Sept. 29. F-Nov. I. J-Nov. 29. N-Dec. 8, C-Oct. 3- G-Nov. 22. K-Dec. I. O-Dec. II, D-Oct. IG. H-Nov. 24. L-Dec. 4- P-Dec. R-Dec. 13 19 III. DISCUSSION OF RESULTS Water. From the tables it will be seen that, beginning with lot L, there is an increase in content of water. The fish in lots A to D were flukes or summer flounders, while those in lots L to R were winter flounders. Apparently the water-content of the winter flounder is about 3 per cent. higher than that of the summer variety. Williams finds the water-content of the plaice (Plenronectes pla- tessa) to be 79.86 per cent., while in the report of the U. S. Com- missioner of Fish and Fisheries the value for the same fish is 77.39 per cent. The government analyst also reports 85.04 per cent. of water for the summer flounder and 84.35 P^r cent. for the winter 64 Cold-Storage of Fish [Oct. flounder {Pleiironectes americanns) . It will be seen that the gov- ernment figures accord neither with Williams' nor our own. The government values are based lipon the analysis of but one fish in each case. Ulrich did not analyze a fresh specimen, but reports the water-content of smoked flounder to be 71.66 per cent. On comparing the results for the three series, it will be noted that there was practically no change in the water-content. This would be naturally expected, because of the care taken to keep the fish completely encased in ice during the storage period. In the case of beef, however, Emmett and Grindley^^ report a loss of 1.3 per cent. of moisture after a forty-three days' period of storage. Total solids. As the value for total solids was determined by difference, it varied inversely as the water-content. Our results show that there was no change in the water-content. Consequently, the value for total solids remained unchanged. Inorganic matter. There was no reason to believe that pos- sible changes in the flesh during cold-storage would affect the ash- yield, yet for an adequate analysis of the fresh specimen it was desirable to make this determination. By repeating the determina- tion on the cold-stored products we were able to obtain a closer value for the ash-yield and at the same time detect any unexpected change. Ash determinations were made only upon summer flounders. The government analyst in the report already quoted gives the following percentage values for the ash content of three related fish : summer flounder, 1.29; winter flounder, 1.20; and plaice, 1.46. As was expected, the results obtained by us showed that cold-storage was without effect on the yield of ash. Organic matter. As in the case of total solids, organic matter was determined by difference. Its variations were dependent upon variations in water-content and ash-yield. There being practically no variations in these, the percentage of organic matter remained unchanged. Ammonium nitrogen. Without doubt this was our most im- portant determination, especially from the Standpoint of detection of bacterial influences. Our method, as already described, differs from that used by Pennington and Greenlee^^ in that we substituted 15 Emmett and Grindley : 7. Ind. Eng. Chetn., 1909, i, p. 413. 18 Pennington and Greenlee: Journ. Am. Chetn. Soc, 1911, xxxii, p. 561. 1913] Clayfon S. Smith 65 sodium hydroxid for sodium carbonate. After a long series of experiments, Pennington and Greenlee found that the results ob- tained with the modified Folin method agreed very well with those obtained when the older magnesium oxid method was used. By using sodium hydroxid we were open to the criticism that our results might be higher than actual ammonium values. Yet granting this possibility, we found that the proportionof ammonium nitrogen was very low, even after a nine months' period of storage. If, therefore, by using a method which might give high results, no in- crease in ammonium nitrogen was found, after six months of storage and only a trivial increase after nine months of storage, it is fair to conclude that the fish were practically unchanged at the end of the last named period of storage. [See the preceding paper by Shulansky and Gies (p. 45) and the succeeding one by Perlzweig and Gies (p. 69).] Pennington and Greenlee found the ammonium nitrogen content of fresh chicken meat to be 0.012 per cent. Houghton^'^ reports 0.021 per Cent, of ammonium nitrogen in fresh light chicken meat and 0.039 P^^ Cent, in the same kind of meat after a period of five months of storage. For dark chicken meat he reports 0.019 P^^ cent. ammonium nitrogen in the fresh sample and 0.026 per cent. after a period of five months of storage. It would appear, then, that so far as the production of ammonium nitrogen is concerned, fish in cold-storage change more slowly than chickens. Total nitrogen. Unless some ammonia was formed in the fish, and escaped into the air, there would be no chance for any diminu- tion in the nitrogen content. Total nitrogen remained unchanged. It will be noticed that there is a diflference of about i per cent. be- tween the nitrogen content of flukes and flounders, This Variation is partly explained when one takes into consideration the difference in Contents of water. " Soluble nitrogen." If any hydrolytic changes took place during the cold-storage period it would be natural to suppose that one or more of the nitrogenous constituents of the muscle became more soluble, or, in other words, that there was an increase in the "soluble nitrogen." Our results indicate that there was no in- i'^Houghton: /. Ind. Eng. Chem., 191 1, iii, p. 497. 66 Cold-Storage of Fish [Oct. creased solubility of nitrogeneous substances. In the case of chicken, Hoiighton reports a slight increase in "soluble nitrogen" for light meat and a slight decrease for dark meat. " Coagulable nitrogen " and " non-coagulable nitrogen." From the data in the tables it appears that there was a very slight increase in " coagulable nitrogen " and a corresponding decrease in "non-coagulable nitrogen." These dififerences are too slight to Warrant any inferences. Lipins. The term lipins is used to indicate the fats and fat-like substances in the "ethereal extract."^^ The flesh of the winter flounders that were used for the determinations was comparatively poor in ether-soluble constituents. The actual weight of lipins neu- tralized at any time was always less than one gram. This admits of relatively large degrees of experimental error; according to Allen, ^^ 5 to 50 gm. of material should be used for this determina- tion. Other authors recommend at least 4-5 gm. In our work such large samples were not available. While the values here reported are possibly somewhat too high, it is significant that there is no increase in acidity during a nine months' period of storage. It is difficult to believe that the lipins would undergoany changes, significant of deterioration in nutritive value, which would not be shown more strikingly by the protein constituents of the flesh. The negative findings in this particular connection accord with such a view of the matter. Reducing substances. We determined the reducing power of aqueous extracts, in order to detect any sugar which might have resulted from the hydrolysis of glycogen during the cold-storage period. A similar determination was made by Williams,^*^ but her method involved the hydrolysis of all carbohydrate-yielding sub- stances. She treated fish-powder with boiling dilute hydrochloric acid Solution under a reflux condenser for three hours; proteins were removed by precipitation with lead acetate ; then, after removal of the excess of lead, the reducing power of the filtrate was de- termined by the Fehling method and reported as percent. of glucose. 18 Rosenbloom and Gies: Biochemical Bulletin, 1911, i, p. 51. 18 Allen : Com. Organic Anal. (3 ed.). Vol. 2, pt. i, p. 105. 20 Williams: Trans. Chem. Soc, 1897, Ixxi, p. 651. I9I3] Clayton S. Smith 67 By this method a reducing power of 2.32 per cent. was reported for the plaice (Pleuronectes platessa). Our own method failed to show any reducing power. Artificial hydrolysis was avoided. We were unable to get satisfactory responses to our qualitative tests for the presence of sugar in protein-free extracts, both from fresh and storage fish. For a com- parative test, glucose, to the amount of o.oi per cent., was added to one of the extracts, and a characteristic reduction obtained. For the quantitative determination of small amounts of sugar in meat, Bauer^^ recommends a spectroscopic method, since the Polarimetrie and titrimetric processes are unreliable for amounts less than 0.5 per cent. As no sugar was detected qualitatively, either before or after cold-storage, it seemed quite certain that no appreciable hydrolysis of glycogen had taken place. Quantitative reaction of aqueous extracts. The reaction of aqueous extracts was always found to be acid to both litmus and Phenolphthalein. From the data in the tables it is evident that the acidity of the aqueous extract was not materially affected by long periods of cold-storage. In several instances the same extract was titrated after Standing for from two to four days in an ordinary house refrigerator. In each case the Variation in acidity was within the Hmits of error of the method itself. The general analytic results of our work are summarized in Table 4. TABLE 4 Summary of general average data Summer flounder (fluke) Winter flounder. Menth s in Water N itrogen Ash Water Nitrogen Lipins cold storage Total Am- monium 5« Total Soluble 5« Non-coag- ulable 1k Per Cent Acidity Nene Six Nine 78.56 78.69 78.45 3-44 3-33 3-40 0.0195 0.0142 0.0213 1.28 1.29 1.28 81.75 81.58 82.04 2.89 2.79 2.83 0.964 0.887 0.937 0.383 0.351 0.353 0.379 0.366 0.443 136 136 127 21 Bauer: Arb. a. d. kais. Gesundhsmt, 1908, xxx, p. 63. 68 Cold-Storage of Fish [Oct IV. SUMMARY OF CONCLUSIONS 1. The proportions of water in, and yield of ash from, the flesh of flounders were unaffected by a nine months' period of cold- storage. 2. The changes in the proportions of soluble, coagulable and non-coagulable nitrogenous constituents were negligible. 3. During a nine months' period of cold-storage, there was prac- tically no change in the content of ammonium nitrogen. 4. For fish with a low content of lipins, there was apparently no increase in the acidity of the muscle lipins during a nine months' period of cold-storage. 5. There was no production of reducing substance from any constituent of the flesh during any of the storage periods. 6. There was no evidence, whatever, of any depreciation in the nutritive value, or any change in the sanitary character, of the fish at any time during nine months of cold-storage. The writer wishes to record his indebtedness and to express his thanks to Prof. William J. Gies under whose direction this work was done.^^ 22 See the succeeding paper by Perlzweig and Gies (p. 69), for additional data on this subject. A FURTHER STUDY OF THE CHEMICAL COMPOSI- TION AND NUTRITIVE VALUE OF FISH SUBJECTED TO PROLONGED PERIODS OF COLD STORAGE i WILLIAM A. PERLZWEIG and WILLIAM J. GIES (Biochemical Laboratory of Columbia University, at the College of Physicians and Surgeons, New York) The study described in the preceding paper* was interrupted in October, 1912, by Dr. Smith's appointment to his present position in the Bureau of Chemistry at Washington. It has given us pleasure to proceed with the work to the end of the second year of storage. As the experiments by Dr. Smith were conducted under the guid- ance of the senior author, it has been a simple matter to continue the study without deviation from the plans and procedures of the preHminary part of the work. The methods of analysis, as well as the detailed conduct of the work, were strictly in accord with the descriptions at page 57 of Dr. Smith's paper. The averages of our analytic data are recorded in the accompanying table, which, for convenience of comparison, includes the analogous figures from Dr. Smith's table on page 6y. The data in the accompanying table indicate very clearly that the fish under examination did not undergo any chemical change of importance, from the standpoint of nutritive value, at any time dur- ing a storage period of two years. General microscopic examination of the flesh indicated that there had been no material alteration of the fibers in any instance. Crystals of triple phosphate were never detected in or on any of the muscle fibers, a finding in accord with the data for ammonium nitro- gen in the flesh. Kept in an ordinary refrigerator after their de- livery to us, these fish, like f resh ones, appreciably deteriorated in a few days and crystals of ammonio-magnesium phosphate could then 1 Smith : Biochemical Bulletin, 1913, iii, p. 54. 69 70 Cold-Storage of Fish [Oct. be detected in the flesh at all exposed surfaces. The whole fish and all portions of the flesh, in every instance (12 hours after thawing), were devoid of any odor that might have indicated significant bac- terial change (comparisons were made with fresh fish of the same kind). The stomachs of the fish, even at the end of two years of General average data pertaining to the composition of flounders in cold storage A. Average analytic data quoted from Smith's paper, p. 67 (Table 4) Summer flounder (fluke) Winter flounder MoDths in cold Water Ni trogen Ash Water Nitrogen Lipins Reac- tion of aqueous extractf storage Total Am- monium Total Soluble Non-co- agulable Per Cent. Acid- ity* 6 9 % 78.56 78.69 78.45 % 3-44 3-33 3-40 % 0.0195 0.0142 0.0213 % 1.28 1.29 1.28 % 81-75 81.58 82.04 % 2.89 2.79 2.83 % 0.964 0.887 0.937 % 0.383 0.351 0.353 0.379 0.366 0.443 136 136 127 1-37 I.17 1-23 B. Averages of our own analytic data 17 78.91 3-46 0.0177 1-23 20 79-14 0.41 1 134 1.28 21 79.24 3-51 0.0205 1.26 22 76-93 81.26 76.32 3-39 0.876 0.433 1.20 23 79.19 75-74 3-26 3.28 0.0231 0.0217 1-25 0-301 147 24 3-55 1.008 0.379 I.'?2 storage, sometimes contained large masses of undigested matter — in a number of instances small fish that had been swallowed were almost wholly undigested!^ The gastric and intestinal membranes were intact, and withstood a surprising degree of tension. The ab- dominal viscera in general were sound and, when handled, emitted no odor that is not common to them when exposed in fresh flounders. The constancy in the data for the yield of ammonium nitrogen, ♦Expressed as mg. of KOH required to neutralize i gm. of the total lipin mixture. t Expressed as c.c. of n/s NaOH sol. required to neutralize 100 c.c. of extract. These data for the first three records {A) were compiled from Tables 1-3 in Dr. Smith's paper. 2 In one case, 2 years and 19 days after the flounder was sent to storage, the stomach contained two fishes, each of which was about 5 in. long and 1% in. wide at the middle of the body. Digestion was advanced at the caudal end of one of these fishes, in the abdominal region of the other, and it was apparent that digestion of the skin had occurred here and there, but it was surprising to note how little change had occurred. There was no putrefactive odor. I9I3] William A. Perlzweig and William J. Gies 71 for the reaction of the aqueous extracts, and for acidity of the lipins, show conclusively that there was no appreciable alteration of the flesh of the fish through bacterial influences. The uniformity in the data for "soluble" and for " non-coagulable " nitrogen (making due allowance for the gradual loss of water from most of the fish as the storage period lengthened) shows that there were no appre- ciable autolytic changes. Some of the fish that had been subjected to analysis, includlng three in storage for two years, were served with meals in conven- tional ways to a number of people, the authors among them. These portions were palatable and entirely acceptable. The taste was sHghtly dififerent, perhaps somewhat more " fishy," though not unpleasantly so, but otherwise there was nothing to suggest a lack of freshness. The data in Dr. Smith's paper and this one pertain to floun- ders that were sent to cold storage very soon after the fish had been caught. These fish were not removed from cold storage before our Order was given for their shipment to this laboratory. They were delivered within an hour afterward, and analysis was begun within 12 hours after their delivery to us, We do not suggest that our findings would apply in any degree to fish that were not strictly fresh and unspoiled before they were put in cold storage. It is obvious, also, that these results have no bearing on the condition of fish which have been removed from cold storage and kept a week or more in a shop, exposed, until sold, to public inspection during market hours, and iced or kept in a common refrigerator at night. It is equally obvious that these data have no material bearing on the cold storage of anything except fish. The results of our studies convince us that fresh fish, similar in general character to flounders, may be preserved frozen, by the best cold storage processes, for at least two years without undergoing any important chemical alteration, and without materially depreciating in nutritive value. THE INFLUENCE OF CHRONIC UNDERNUTRITION ON METABOLISM Prof. N. Ziintz recently reported^ the results of an investigation which the writcr of this note undertook under his personal guidance at the Tierphysiologisches Institut, Berlin, and which was later con- tinued by Dr. Diakow of St. Petersburg. The subject of these experiments, a female mongrel weighing lo kg., had been kept on a controlled diet ( 1 50 gm. of horse meat and 80 gm. of rice) for nearly a month, during which period three res- piration experiments were performed with the Regnault-Reiset appa- ratus. Although the food was not quite sufficient to completely Cover the energy requirement of this dog, the latter had been putting on nitrogen, but towards the end of the preliminary period a State of equilibrium was attained. Beginning February 23, 1912, the diet had been considerably reduced (60 gm. of meat and 35 gm. of rice) and for over a year, until the animal's death, it remained below the actual need of the organism. A respiration experiment, performed about three weeks after the insufficient feeding had been begun, showed a shortage of 220 cal. per day, which had been drawn f rom the dog's substance. Con- sidering that, as the nitrogen determination in the urines showed, only 7.5 cal. had been derived from the protein material (corre- sponding to 9 gm. of flesh), the remaining 212.5 cal. must have been furnished by 22.4 gm. from the fat depot of the organism. The total metabolism had changed, in the meantime, from 553 cal. to 394 cal. per day. The limits of this review will not permit discussion of each ex- periment. The average number of cal. liberated by our dog, in the course of the subsequent six weeks, was 149 cal. in excess of the supply per day. As there had been very little loss of nitrogen, it may be assumed that all the extra energy was derived from body 1 Zuntz (Versuche von S. Morgulis and M. Diakow), Einfluss chronischer Unternährung auf den Stoffwechsel : Biochemische Zeitschrift, 1913, Iv, p. 341. 72 1913] Sergius Morgulis 73 fat, in which case a loss of 770 gm. should have been occasioned. In other words, the body weight would have changed from 7.53 kg, to 6.76 kg.; the weight actually observed at that time was 6.96 kg. Likewise, on the 20th of July, when the body weight had diminished to 6.36 kg., the observed loss fully agreed with the expectation based upon resuhs of the respiration experiments. After I left Beriin, the dog was maintained on the same diet and, in January 1913, my colleague, Dr. Diakow, continued the series of experiments which had been begun a year ago. In the concluding period of 82 days, i. e., until the animal died, Dr. Diakow per- formed five respiration experiments. The excess of energy pro- diiced has been 117 cal. per day, which is equivalent to 11.7 gm. of fat. Since 1,400 gm. were lost during that period, or 17.1 gm. per day, it is evident that some of that energy must have been derived from protein. Unfortunately, Dr. Diakow made no determinations of the total nitrogen in the food and in the urines, but Professor Zuntz computes that an average of at least 0.2 gm. must have been lost daily. The most remarkable fact in connection with the second part of this research is the rise in the energy requirement which, in spite of the fact that the body temperature was very low, increased to the same level as before the underfeeding. Professor Zuntz's conjec- ture that this rise in the metabolism was coincident with an increased nitrogen elimination, as the fat was being exhausted, is entirely borne out by the results of my recent unpublished work on the metabolism of chronic underfeeding wherein this matter has been studied from a much wider point of view. Sergius Morgulis College of Physicians and Surgeons, New York NITROGEN METABOLISM DURING CHRONIC UN- DERFEEDING AND SUBSEQUENT REALIMENTATION In the course of an extended physiological investigation of the effects of chronic underfeeding, a number of observations on the nitrogen metabolism have been made which will be briefly discussed in this preliminury note. The experiments were made with a dog weighing 14 kg. The animal was fed a very liberal, but not excessive, diet for ten days. During this time it was found that 78.7 percent of the absorbed nitrogen had been eliminated by the kidneys. The quantity of food was then reduced to one-third, so that in the course of several weeks the dog was obliged to live at the expense of its own tissues. The amount of nitrogen in the food was very low, which circumstance caused a continuous waste of the nitrogenous material of the body. Correspondingly, the nitrogen eliminated in the urine for the first week of underfeeding was 85.4 percent above the amount actually absorbed. The supply of nitrogen during the period of chronic starvation varied from 21.7 to 23.1 gm., per w-eek. The nitrogen elimination remained very uniform until a late period in the experiment, when quite abruptly it increased about 25 percent. This increased nitro- gen elimination coincided with a series of interesting changes in the animal which cannot, however, be alluded to here. It was occa- sioned by a greater combustion of protein, due to the exhaustion of the fat of the body. The specific gravity of the daily urines throws some light on this matter. The average sp. gr. of the normal urine was 1.0141, while during the early part of the underfeeding it was 1.0124. In the concluding two weeks the sp. gr. rose to 1.0186 (the volume being practically the same), owing to the liberation of inorganic substances by the protein decomposition. The study of the gaseous exchange and the respiratory quotients furnished fur- ther unequivocal proof that the organism derived its energy from the oxidation of the muscular substance. After the animal's weight had diminished over 40 percent, and its physical strength had been reduced to such an extent that it was unable to enter its cage but had to be lifted into it, the dog was again given a rieh diet ; and in f our weeks time it f ully recovered the 74 1913] Sergius MorguHs 75 original weight. The general physiological transformation of the animal during that short time has been studied carefully and will be related in detail when the entire work is published. Nitrogen was avariciously retained by the organism during realimentation, so that in these four weeks the organism was en- riched by 191 gm. of protein after it reclaimed what had been lost during the chronic underfeeding. In the first week only 58 percent of the absorbed nitrogen appeared in the urines, but this relation gradually changed; and as early as the fourth week, 73.8 percent was eliminated, or nearly as much as for the preliminary period. With the resumption of feeding a striking change occurred in the reaction of the urine. The urines were tested every day and were invariably acid to litmus paper. On the third day of refeed- ing with superabundant quantities of food, it was observed that the urine lost its clearness, having become strongly alkaline. It seemed at first that the alkalinity was caused by bacterial contamination, but preservation of the urine with thymol over night did not prevent its being alkaline. There was no indication of disease in the dog, although persistent alkalinity of the urine is generally regarded as a Symptom of cystitis. The alkalinity was due to an excess of ammonium carbonate in the urine, as could be shown by a very simple experiment. Two different strips of litmus paper were dipped in the urine, whereupon the red became dark blue, but the blue Strip remained unaffected. The two strips were then allowed to dry, when the red color was restored to the former and the latter turned red. The strong ammoniacal smell of the urines left no doubt as to the true cause of the alkalinity. This condition lasted for only a f ew days, when the urines cleared up again ; and the nor- mal acid reaction returned, and remained undiminished thereafter. Unfortunately, I was unable to study the nitrogen partition, but I would venture to suggest that the great influx of phosphates and acid cleavage products of the protein digestion, coupled with a generally impaired condition of the liver and of the whole organism, for that matter, resulted in a rapid elimination of ammonium car- bonate before its transformation into urea. Sergius Morgulis College of Physicians and Surgeons, New York. PROCEEDINGS OF THE BIOLOGICAL SECTION OF THE AMERICAN CHEMICAL SOCIETY, ROCHESTER, NEW YORK, SEPTEMBER 10-12, 1913 I. EXECUTIVE PROCEEDINGS The Section met with the secretary acting as the chairman at the Session of the first day. Announcements were made to the effect that the chairman, Dr. C. L. Aisberg, was in attendance at the meeting of the American Public Health Association, at Colorado Springs and hence would not be present at the first day's session of the Section ; that the Council of the Society had approved the by- laws as proposed by the committee for the proposed formation of the Section into a Division ;^ that a copy of the by-laws, which would be presented for consideration and final action at the last session, was available for inspection ; and that a nominating committee con- sisting of P. Rudnick, M. X. Sullivan, and P. A. Kober had been appointed. The Section then proceeded with the reading of papers, of which abstracts are given on pages 80-95. Following the reading of papers, the chairman addressed the Section. (See page y].^ The session closed with the business meeting. The by-laws, as recommended by the committee and approved by the Council, werc unanimously adopted. The nominating committee reported their recommendations : Chairman, C. L. Aisberg; vice-chairman and secretary, 1. K. Phelps; members of the executive committee — W. D. Bancroft, chairman, Edward Kremers, A. W. Dox, A. D. Emmett, and D. D. Van Slyke. The gentlemen nominated were unanimously elected the officers for the ensuing year. The secre- tary read a Statement drawn up by W. A. Noyes concerning the advisability of the Division taking some action toward securing a special Journal restricted to organic and biological papers, and pub- 1 Aisberg: Biochemical Bulletin, 1911, i, p. 94. 76 I9I3] I- K. Phelps 77 lished by the Society. Considerable interest in the possibility of such a Journal was shown, but, as no practical method of financing the project was presented, no action was taken. I. K. Phelps, Secretary. Bureau of Chemistry, U. S. Department of Agriculture, Washington, D. C. II. CHAIRMAN'S ADDRESS, SEPTEMBER 12, 1913 Gentlemen, I did not come to Rochester with the intention of making a speech, but find — I am sorry to say — that Professor Cham- bers expects me to talk. He made the request — or, shall I say, demand — as we came into this room. I find that I am driven to the usual refuge of those who have to speak when they would rather be silent — that is, I will take refuge in the history of my subject. This subject has, I think, some general interest because originally no very definite distinction was made between biochemistry and any other kind of chemistry. One of the first real biochemists was Lavoisier, whom all matter, whether living or dead, interested. He performed the first calorimetric experiments. He was the inventor of the ice calorimeter, and showed that animal heat was the result of oxidation. All the chemists of that generation and the imme- diately succeeding one did biochemical work. I need only cite Liebig, who is perhaps in some ways the greatest of all biochemists. Unfortunately, about the latter part of Liebig's life chemists lost in- terest in biochemistry. This was due very largely to the sudden and tremendous development of organic chemistry, which was brought about by the discoveries of men like Hof mann and Kekule. It was so easy to make new synthetic substances and, thereby, gain a sort of immortality, even though the main result of putting a chlorine atom here and a bromine atom there was to fill up Beilstein. In consequence, thoroughly trained chemists did not busy themselves with subjects that w^ere really important in the elucidation of that matter which is found in living organisms, and which forms the physiological basis of life. The scientists in biology and medicine needed such Information. The chemists did not glve it to them. Consequently, physicians and physiologists who were ill-equipped for chemical research were forced to carry forward the work of 78 Biological Section, American Chemical Society [Oct biochemistry. Though the net result of their work made decidedly for progress, only too often it created confusion and artificial diffi- ciilties. Even the best biochemists of those days make us wonder why they did not pursiie their chemical investigations as far as the chemical methods of that day would permit. The answer is, I think, in many cases, that they were not real chemists but physiolo- gists with a chemical veneer. Fortunately, this has been changing during the past decade, largely owing to the work of Emil Fischer. While we recognize in him a master of chemical technique, we may be certain that in a measure, at any rate, the preeminent position which he occupies among the chemists of his time is due to his clear conception of the really most important work in organic chemistry along biochemical lines. Fortunately, more and more organic chemists are following in his footsteps, and are devoting their at- tention to substances which occur in living things. I wish here to make a plea for more of this sort of work in America. I believe that the rewards and recognition for knowledge of chemistry applied in biochemistry are great, because the work of the biochemist will be applauded not merely by chemists, but also by zoologists, botanists and physicians. A biochemist has a wider audience because his work presents a more general appeal than the work of organic chemists upon such subjects as dye-stufifs and the like. Further, I wish to point out the value of Instruction in allied subjects. Not every organic chemist can successfully attack all biochemical problems. Besides his organic chemistr}^ other expe- rience in physiology and, above all, experience in dealing with substances which do not crystallize, is necessary. In many cases it is difificult to conduct biochemical research because the bio- chemist must very frequently begin with the smears which the organic chemist consigns preferably to the slop jar. While the things which will not crystallize interest less the organic chemist, they are the very classes of substances with which the biochemist must deal. Great care, great patience and a knowledge of colloids are required of the organic chemist who wishes to work in biochem- istry, but I feel confident that the reward for such men is great, not merely in pure science, but also in industries and in the arts. 1913] Carl L. Aisher g 79 The history of biochemistry in America is similar to that abroad. In America it developed first in the seventies and eighties in the medical schools of the country; and, at that time, it was controlled by physicians and physiologists abroad. The subject was narrowed to the consideration of biochemistry as affecting the Hfe of man; that is to say, the chemical side of physiological processes of the human body together with such considerations of bacteriological chemistry as affect man in health and in disease. This phase of biochemistry is cared for very adequately and acceptably by the American Society of Biological Chemists, the first biochemical Society to be formed in America. The phases of biochemistry which the American Chemical Soci- ety can very naturally expect to encourage are quite distinct f rom the aims of the American Society of Biological Chemists. Our useful- ness will include the biochemistry affecting agriculture, phytochem- istry, in particular, and such industrial processes as are based upon biochemical reactions; for example, the more exact study of the chemical composition of fruits, grains, and food products. It must be admitted that, at present, we know only those chemical substances occurring in considerable amounts in such important grains as wheat and corn. The minor constituents in grains of much importance have not been identified with exactness. If we consider grains of less importance, even this degree of knowledge can not be claimed. Some of our most important modern industries, like those deal- ing with starch, artificial fabrics, leather, tanning materials, glue and gelatin, meat packing and the flour milling industry require biochemists, and we are now training men to deal with such practical Problems. If our Society confines itself to the activities already mentioned, there still remains a wide field of biochemistry uncared for: the biochemistry of the lower animals. This part of the biochemical work will become a part of the work in the zoological societies of the country. My view is that three societies of biological chemistry can well exist in America without competing in any way, and each one car- ing for a specific need. These would include the biochemistry of 8o Biological Scction, American Chemical Society [Oct. the higher animals and its application to medicine ; the biochemistry of the lower animals ; and biochemistry in its appHcation to plants, agricuhure, and the industries. Carl L. Alsberg, Chairman Bureau of Chemisiry, U. S. Department of Agriculture, Washington, D. C. III. SCIENTIFIC PROCEEDINGS (ABSTRACTS) On the presence of histidine-like substances in the pituitary gland (posterior lobe). T. B. Aldrich. (Research Laboratory of Parke, Davis & Co., Detroit, Mich.) Employing Pauly's dia- zobenzene sulphonic acid reaction for the detection of histidine, it seems probable that histidine or some form of it is contained in a free State in the desiccated posterior lobe of the pituitary gland since, by benzoylating direct, using Inouye's method, Pauly's reaction was positive. The substance (or substances) giving Pauly's reaction, after hydrolysis by means of mineral acids or digestion with pan- creatin, is not tyrosine (which gives a similar reaction) since, after benzoylating, the histidine reaction still persists. Furthermore, the histidine-like substance (or substances) is probably not histidine, since it does not give Weidel's reaction as modified by Fischer, or Knopp's reaction with bromine. It is probable, also, that Pauly's reaction is not a specific reaction for histidine. The mutual action of pepsin and trypsin. J. H. Long. (Northwestern University Medical School, Chicago.) The earlier physiologists seem to have considered this a comparatively simple question, but their findings were not in agreement. Kühne was one of the first to discuss the problem and he concluded that pepsin destroys trypsin. This is probably correct but his experimental evidence does not Warrant the Statement. In all such experiments the reaction of the medium must be definitely known as the concen- tration of hydrogen or hydroxyl ions is often the determining factor. In most of the earlier work these points were almost wholly over- looked, as the combining power of protein for acid or alkali was either not known or not recognized. Making due allowance for the reaction of the medium, the present experiments show that, within the practical limits of behavior in the body, trypsin has no important I9I3] /• K. Phelps ' 8i action on pepsin, whereas the action of pepsin on trypsin is markedly destructive. While an acid medium weakens trypsin, pepsin plus acid seems to destroy it rapidly. A further study o£ the well water o£ Delaware, Ohio. G. O. HiGLEY, {Ohio Wesleyan University, Delaware, 0.) This study was intended to Supplement the report made at the spring meeting : to trace the relation between well water and an outbreak of typhoid. The city water has been examined and found safe. The water of about 100 wells has been analyzed and much of it found polluted. Five vaults were selected in various parts of the city and in markedly different soils : these were heavily salted and a weekly test f or Chlorides made during a period of nearly two months, in the water of thirteen wells located from 58 to 118 feet from the vaults. Com- parison of results of analyses, made before and after the salting process, showed a decided increase in chlorides in the well water at four of the five centers and in seven of the thirteen wells. On the distribution of mercury following acute bichloride o£ mercury poisoning. Jacob Rosenbloom. (Biochemical Lab- oratory of the Western Pennsylvania Hospital, Pittshnrgh, Pa.) Estimations of the amount of mercury in the organs of a woman who died 8 days after ingestion of bichloride of mercury. The non-interference of ptomaines with certain tests for morphine. Jacob Rosenbloom and S. Roy Mills. (Biochemical Lahoratory of the Western Pennsylvania Hospital, Pittsburgh, Pa. ) We have determined experimentally that bacterial products, formed during aerobic and anaerobic putrefaction of various human organs, do not give reactions simulating those due to the presence of mor- phine. In no way do they interfere with the detection of morphine when the latter is added to a mixture of these putrefactive products. The effect of electrolysis on whole proteins, Witte peptone, and some of their decomposition products. James P. Atkin- SON. (Chemical Lahoratory, Department of Health, City of New York.) Whole protein (egg white), Witte peptone, and protein (horse serum) hydrolyzed by hydrochloric acid, yield approxi- mately 50 percent of the total nitrogen as ammonia, when electro- lyzed in a sulfuric acid Solution. The amino acids tested, glycyl- glycine, uric acid and urea, do not yield as much nitrogen in the 82 Biological Section, American Chemical Society [Oct. form of ammonia nnder the same conditions. Ammonium sulfate is unaffected. The non-development of cytolytic sera following the intra- venous injection of mould spores. A. F. Blakeslee and R. A. GoRTNER. {The Carnegie Institution of Washington.) Intraven- ous injections of the spores of each race of Miicor "V" were given to rabbits, rabbit No. 5 receiving 30 injections of the J* race and rabbit No. 55 receiving 29 injections of the ? race. Each injection averaged about 500,000,000 spores. Following the last injection of approximately 800,000,000 spores, a loop of blood was taken at intervals of 30 minutes for 6 hours, then every hour for 4 hours more, then every 2 hours for 16 hours more, and later at less fre- quent intervals. Separation cultures were made of agar which contained the loop of blood taken and the number of mould colonies which developed were counted. A similar test was made at the same time, using rabbits which had received their first injection of the spores. In each case the disappearance of the spores occurred after about 43 hours, the immunized rabbits retaining the viable spores as long as the control rabbits. Effect of acids upon the catalase of taka-diastase. Ray E. Neidig. {Chemical Section of the Iowa Agriciiltural Experiment Station, Ames.) Data were presented showing the inhibiting effect of several of the important inorganic and organic acids on the action of the catalase of taka-diastase. Curves were plotted, for different acid concentrations, which show the quantity of oxygen liberated at stated intervals. The acids, arranged in order of the magnitude of their inhibiting effect for equi-normal Solutions, are as f ollows : sulfuric, h3^drochloric, oxalic, tartaric, citric and acetic. The inhibiting effect of the first three was much more pronounced than that of the others. Neutralization of the acid Solution usually restored some of the activity, the amount of increase depending upon the particular acid used. Van Slyke's amino-nitrogen appa- ratus was used in these experiments for measuring the amount of oxygen liberated. Polyatomic alcohols as sources of carbon for molds.^ Ray E. Neidig. {Chemical Section of the Iowa Agricultural Experi- 2 Neidig : Jour. Biol. Chem., 1913, xvi, p. 143. I9I3] I- K. Phelps 83 ment Station, Arnes.) A comparison of some of the polyatomic alcohols occurring in nature was undertaken in order to determine the degree of their utilization by molds as sole sources of carbon. The alcohols used were methyl alcohol, glycol, glycerol, erythrite, adonite, mannite, dulcite and sorbite. Eight species of molds rep- resenting four genera were cultivated in media containing these alcohols. It was found that methyl alcohol produced no growth, glycol induced germination only, glycerol produced strong cultures, erythrite was utilized by the majority of molds and adonite by only a few, while all three of the hexatomic alcohols may be regarded as good sources of carbon. These results indicate that molds are able to use both optically active and inactive Compounds as sources of carbon. If viewed from the Standpoint of their oxidation products, it is possible that active Compounds are first formed and these are then utilized in the development of the molds. Cleavage of benzoylalanine by mold enzymes. Arthur W. Dox and W. E. Ruth. {Chemical Section of the Iowa Agricul- tural Experiment Station, Arnes.) See page 23. Influence of certain organic substances upon the secretion of diastase by various fungi. Christine Chapman and W. C. Etheridge. (Laboratory of Plant Physiology, State College of Agricultnre, Cornell University, Ithaca, N. Y.) In this work the influence of varying concentration of cane sugar, glucose, peptone and tannic acid upon the secretion of diastase by Aspergillus niger, Aspergillus oryzae, Penicillium expansum, Penicillium camembertii, Mucor Rouxii and Cephalothecimn roseum has been investigated. Czapek's Solution was employed with the sugar replaced by 0.4 per- cent soluble starch. To this was added the substance whose influ- ence was to be determined. It was found in general that the pres- ence of any of these organic substances retarded the secretion of diastase by the fungi mentioned. The higher the concentration the greater the retardation. A method for studying slight degrees of glycosuria, adapted from Macleod and S. R. Benedict. Amos W. Peters and Mary E. Turnbull. (Biochemical Laboratory, Training School for Feeble-minded Children, Vineland, N. J.) Urine is clarified by the method of Macleod, i. e., urine + concentrated acetic acid + 84 Biological Scction, American Chemical Society [Oct. Merck's blood charcoal. No sugar is lost by this procedura. I£ the urine is diluted to only 7/5 original volume, the filtrate is water-clear for polarization. Of the filtrate, 5 c.c. are transferred to a 100 c.c. Kjeldahl flask, neutralized with a saturated Solution of sodium car- bonate, using alizarine as the indicator, and 5 c.c. of a modified Benedict reagent are added. After placing a pebble in the liquid and fixing the flask in an inclined position directly over a small Bun- sen flame, the whole is boiled for 2^ minutes. The resulting small volume is transferred to a centrifuge tube and made up to 10 c.c. Examined under a shaded electric light and against a dark background, even a trace of glucose shows turbidity and after centri- fugation as little as 0.0035 percent shows a film of red precipitate. Quantitative estimations are made by comparison with Standards based upon a normal urine excreted on a normal diet and showing zero rotation (or nearly so) after clarification, and to which glucose is added in amounts that increase the content by successive incre- ments of 0.0 1 percent. The sensitiveness is so great that such slight differences in proportion may easily be detected. Composition of the modified Benedict reagent : sodium citrate, 100 gm.; sodium acetate, 100 gm.; sodium carbonate (anhyd.), 50 gm.; cryst. copper sulfate (Kahl.), 12.5 gm,; water (dist.), 500 c.c. The estimation of protein and amino- and nucleic-acids in potable waters. Philip A. Kober. (Harriman Research Laho- ratory, Roosevelt Hospital, Nezv York.) Experiments show that by using the right precipitants and evaporating to one-tenth of the original volume, proteins and nucleic acids can be estimated in pot- able waters by the author's nephelometric method. This method will easily reveal the presence of one part of substance in one million parts of water. By using the copper method,^ potable waters may be analyzed for amino-acid nitrogen before or after hydrolysis. This method will reveal one part of amino-acid nitro- gen in one million of water, without difficulty. The fate of protein digestion products in the body. Donald D. Van Slyke and Gustave M. Meyer. (Rockefeller Institute for Medical Research, New York.) Previous work by the authors 3 Kober: Jour. Amer. Chem. Soc, 1913, xxxv, p. 1546. I9I3] I' K. Phelps 85 has shown that during digestion amino acids are absorbed into the blood, as the amino-acid nitrogen of the latter, per 100 c.c, rises, in a dog, from 4-5 mg. before feeding to 10—12 mg. after a meal of meat. The low concentration of amino acids in the blood even at its maximum indicates that the digestive products must be removed rapidly from the circulation. This is found to be the case after the injection of amino acids directly into the circulation. They dis- appear from the blood almost as fast as they enter it. Analysis of the tissues shows that these have absorbed the amino acids from the blood, without subjecting them to any immediate chemical change. This apparently follows later, but in the muscles the change is so slow that no decrease in amino-acid nitrogen can be determined within the first 3-4 hours after the injection. In the liver, on the other band, the amino acids absorbed as the result of the injection have entirely disappeared by this time, indicating that the metabol- ism of these products is particularly rapid in the liver. It is less so in the other organs, but whether as sluggish as in the muscles is not yet certain. During starvation the amino nitrogen of the tis- sues, which amounts to 40-80 mg. per 100 g. of fresh tissue, tends to increase rather than disappear, indicating that the amino acids of the tissues can originate from autolysis of the tissues themselves as well as from digestion of food proteins. The configuration of some heptoses. George Peirce. (Laboratory of Pharmacology and Toxicology, University of Wis- consin.) J-a-Mannohexahydroxyheptoic acid and fl?-a-galahexa- hydroxyheptoic acid yield, on oxidation, two pentahydroxypimelic acids that are optical antipodes of each other. The configuration of four of the asymmetric carbon atoms in each monobasic acid is known and the configuration of the fifth is indicated by the above fact. The corresponding heptites are also optical antipodes. Of the four configurations on page 86, I and III are seen to be the two that give optical antipodes on oxidation or reduction of the end carbon atoms. These two are therefore the formulae for the a-com- pounds. The /3-galactose Compounds of formula IV have been syn- thesized but the /?-mannose Compounds of formula II have not yet been prepared. 86 Biological Scction, American Chemical Society [Oct. COoH CO2H CO,H CO2H HCOH HOCH HCOH HOCH HOCH HOCH HCOH HCOH CH.OH HOCH HOCH HCOH HCOH CH.OH HCOH HOCH HOCH HCOH CO2H HCOH HOCH HOCH HCOH COoH I n From d-Mannose in IV From d-Galactose Vanillin in wheat and its relation to soil. M. X. Sullivan. {Bureau of Solls, U. S. Dep't of Agriculture, Washington, D. C.) By means of the sodium bisulphite aldehyde method, an aldehyde that smelled like vanillin, and gave vanillin color reactions, was f ound in the alcohol and ether extracts of ungerminated wheat seeds ; in the roots, seeds, and tops of young wheat seedlings; in rotten wood ; and in the water in which wheat had germinated and grown. Estimated quantitatively by Polin and Denis' colorimetric method, the amount in the ungerminated seed is small (several parts per million) but is considerably increased during germination and the early stages of growth. Treating the seed with 5 percent sulfuric acid Solution also increased the amount of vanillin that could be extracted. The presence of vanillin in other plants was indicated. The vanillin of soil undoubtedly has its origin in part in vegetable debris and the growing plant. Some organic constituents of the culture Solution and the mycelium of molds from soil. M. X. Sullivan. {Bureau of Soils, U. S. Dep't of Agriculture, Washington, D. C.) Examination was made for the organic constituents of the dried mycelium of mixed mold cultures from soil and of Penicillium glaucum grown on Raulin's Solution, and of the filtered Solution after mold growth. In the mixed molds was found a large number of organic substances, many of which were subsequently detected in P. glaucum. In the I9I3] I- K. Phelps Sy alcoholic soda extract of P. glaiicuni were found oleic and palmitic acids, a fatty acid melting at 54° C, a fatty acid, which appears to be elaidic acid, hypoxanthin, guanin, adenin, histidin, thymin and cholin. In the direct alcohol extract were found mannite, choles- terols, hypoxanthin, and cerebrosides. From mold grown on RauHn's Solution plus peptone a small amount of guanidin was ob- tained. In the culture Solution, after a number of weeks' growth, were found fatty acids, purin bases, a small quantity of a histidin- like substance, pentose sugar, unidentified aldehydes, etc. Many of these Compounds have been found in soil and the conclusion is drawn that microorganisms, such as yeasts, bacteria and molds, play an important part in their formation. A method for the determination of small amounts of fat. W. R. Bloor. {Laboratories of Chemistry of Queens University, Kingston, Canada, and of Biological Chemistry of Washington University, St. Louis, Mo.) The method consists essentially in extracting the fat from the tissue or liquid with an excess of alcohol- ether (25 percent ether), measuring an aliquot portion of the filtered extract into distilled water and determining the amount of fat by comparison of the cloudy Suspension so obtained with a Standard fat Solution by the use of the nephelometer. The method has given good results with blood and milk. Nitrogenous hydrolytic products of several Phosphatids. C. G. MacArthur and G. Norbury. (University of Illinois.) Sheep-brain kephalin, sheep-brain lecithin, ox-heart cuorin and ox- heart lecithin were prepared, purified, and then hydrolyzed in a dilute hydrochloric acid Solution. In each case the fatty acid residue contained nitrogen, usually about one-sixth of the total. The fil- trate nitrogen was separated by a special method into four fractions, representing (i) ammonia, (2) chohn or other basic Compounds, (3) amino acids, or Compounds not precipitated by platinum chloride but precipitated by mercuric acetate in a sodium carbonate Solution, and (4) the filtrate from (3). The two lecithins contain about two- fifths of the nitrogen in form 2, while kephalin and cuorin contain practically none. In all of them, fraction 3 is large, varying from one-third to one-half. Fatty acids from kephalin. L. V. Burton and C. G. Mac- 88 Biological Section, American Chemical Society [Oct. Arthur. ( Univcrsity of Illinois.) The fatty acids obtained from hydrolyzing purified kephalin in a dilute hydrochloric acid Solution were separated by the lead acetate method into the saturated and unsaturated fatty acids. The saturated-acid fraction represented about one-third of the total and was found to contain stearic and palmitic acids in the ratio of three to one. The unsaturated fatty acids were separated by the bromination method into clupanodenic, linolic and oleic acids. The amount of clupanodenic acid present was small — less than 2 percent. The linolic acid was found to rep- resent about one-sixth, oleic acid about one-third, of the total fatty acids. A metabolism experiment with swine. E. B. Forbes. {De- partment of Nutrition, Ohio Agricultural Experiment Station, Wooster.) The usual practical rations for swine contain an excess of acid over basic mineral Clements. Urinary ammonia varies directly with this excess of mineral acid, provided the protein intake remains the same. Increased protein intake increases urinary ammonia. This excess of mineral acid in practical swine rations does not seem to affect calcium retention. Water drinking causes the elimination of sodium and chlorin; abstinence from drinking leads to their retention. The feces may contain an abundance of sodium but are nearly free from chlorin. Potassium, magnesium and chlorin balances were usually positive, but were negative during periods of maximum intake, apparently through over-response in the way of protective elimination of the excess ingested. Calcium retention was satisfactory only on rations including meat meal (containing considerable bone) and skim milk. Neither cereals nor soy beans furnish the calcium requisite for growth. An excess of magnesium over calcium caused loss of calcium with a ration of rice polish and wheat bran. The excess of magnesium over calcium in corn and in other practical rations does not appre- ciably restrict calcium retention. The important deficiencies of corn are, in order of magnitude, calcium, phosphorus, and nitrogen. Creatinin elimination was entirely independent of food but varied in the same order as live weight, weight of dressed carcass, of flesh, of bones and of blood. I9I3] /. K. Phelps 89 Soy beans, meat meal and skim milk increase the digestibility of the carbohydrates of the corn with which they are fed. Meat meal and skim milk increase the apparent digestibility of the fat, and decrease the digestibility of the crude fiber of the corn with which they are fed, the results being digestion coefficients of more than 100 and less than nothing. The acidity of normal urine. Howard D. Haskins. (Labor- atory of Physiology and Biochemistryj Western Reserve Medical College, Cleveland, O.) Certain modifications of Henderson's method were suggested. Permanent color Standards were proposed for the ränge of acidity determined by paranitrophen'ol, A report was made of a study of variations of acidity in 24 hour samples and in fractional samples, i. e., the day's urine collected in five periods. No relation of concentration of urine to acidity was noted. The effect of diet was slight. Night urine was distinctly acid in 50 per- cent of the cases, and morning urine (breakfast to 11) was of very low acidity in 50 percent of the cases. Sweating seemed to have a marked effect in causing higher acidity. Sunlight and health. Wilder D. Bancroft, (Cornell Uni- versity, Ithaca, N. Y.) It is usually considered that plenty of sun- light is beneficial to health but Woodruff considers it harmful, espe- cially in the case of tuberculosis. This discrepancy becomes less serious if we consider how light acts. The primary action of light is to tend to eliminate the substance which absorbs it; the primary action is a destroying one. On the other band we get a secondary eüfect with living matter, which is or may be a stimulating one. Strychnine is beneficial in small quantities and toxic in large ones. When studying the effect of light on organisms, one should differen- tiate the two effects. The nature of humus and its relation to plant life. S. L. JoDiDi. {Office of Physiological and Fermentation Investigations, Bureau of Plant Indiistry, U. S. Dep't of Agricultiire, Washington, D. C.) See page 17. The importance of food accessories as shown by rat-feeding experiments. F. C. Cook. (Animal Physiological Lahoratory, Buerau of Chemistry, U. S. Dep't of Agricultiire, Washimgton^ D. C.) Twelve white rats were fed on a basal diet of protein, fat, 90 Biological Scction, American Chemical Society [Oct. and carbohydrate, plus Röhmanii's salt mixture, for a period of 80 days. Most of the rats lost weight during the last three weeks. To the basal ration during 35 days immediately following the 80 days on the basal diet alone, 5 c.c. of a Solution of meat extract, plant extract, or whole milk, were alternately added, the nitrogen and sodium chloride of the three accessories being equal. When meat extract or milk was fed, a marked increase of weight was ob- tained. Eleven young white rats were fed for 35 days on a basal diet, plus one of the three accessories. The stimulating efTect of milk as shown by Hopkins was noted. The meat extract also acted as a stimulant, while the plant extract showed little, if any, stimulat- ing action. The milk and meat extract gave the biuret reaction and heavy precipitates with phosphotungstic acid, but the plant ex- tract did not give either of these reactions. The meat extract, which is a hydrolyzed product practically free from fat and carbohydrate, seems to possess the stimulating properties similar to milk, a natural product. That the calories are not the sole guide in feeding experi- ments, in harmony with the work of Hart, Hopkins, Osborne and Mendel, and others, was noted. The rats gained in weight on a smaller number of calories when milk or meat extract was ingested. No gain in weight was obtained with a larger number of calories in the food ingested in the absence of milk or meat extract. A time recorder for kymograph tracings. Oliver E. Clos- SON. {Research Laboratory of Parke, Davis and Co., Detroit, Mich.) It is at best a tedious Operation to find the projection of the time record on the different graphs as ordinarily traced upon smoked paper. The time interval can easily be recorded by a fine line, entirely across the paper by the following simple device. A fine spring wire stretched two to three mm. from the smoked surface will strike the smoked paper on the rebound and remove a fine line of soot, when picked by the armature of the time-signal magnet. By a little adjustment, a single distinct line is recorded at each closure of the circuit. If it is inconvenient to adjust any recorder to write perpendicularly to the base line, it is a simple matter to make the adjustment so that the time line will be parallel to any such line. Apparatus for studying oxidases. Oliver E. Closson. (Re- 1913] I' K. Phelps 91 search Lahoratory of Parke, Davis and Co., Detroit, Mich.) As the reaction of oxidases with hydrogen peroxide liberates heat, the temperature factor as well as the expansion of the gas are very important and necessitate a thermostat control with continued agitation of the mixture for comparative studies. To obtain uniform temperature and continuous record of the lib- erated gas, the following apparatus was devised. A shaking mem- ber with two compartments, one for holding the hydrogen peroxide and the other for the enzyme Solution, is connected by a tube with ground Joint to a large cylindrical Container with its center at the axis of motion so that liquid in this Container is not agitated by motion around the axis. This arrangement allows the shaking of the reacting Solution and the measuring of the liberated oxygen by the water displaced. The large Container has a tube extending along the axis to the outside of the thermostat, which allows the discharge of the displaced water into a vessel suspended by a spring, so that a writing arm can be made to record the volume, giving on a rotating drum a curve which can be analyzed at one's leisure. Surface tension in muscle contraction. William N. Berg. {Washington, D. C.) Macallum* quotes Jensen^ to the ef^ect that " a thread measuring i mm. in diameter formed of the Plasmodium of Chondrio derma, a Myxomycete, may, when it is in the dense con- dition, bear up a weight of nearly a gram. If the force engaged is surface tension it would amount to about 6000 dynes per centi- meter. . . ." At the same time Macallum does not quote Pfeffer^ who, in dis- cussing the mechanics of ameboid movement says, that in the case of the Plasmodium of Chondrio derma, the outer membrane may vary in its consistence from that of the fluid protoplasm in the interior of the cell, to that of solid gelatinous masses. Chondrioderma have the property of varying the consistence of the outer layer, and Pfeffer regards the tougher outer layer as a physiological product caused by reversible changes in cohesion. Pfeffer further states that not until the outer layer has been brought back to its original 4 Macallum : Jotir. Biol. Chem., 1913, xiv, Proc. Amer. Soc. Biol. Chem., p. xxii. ^ Jensen : Anatomische Hefte, 1905, xxvii, p. 842. 6 Pfeffer : Pflanzenphysiologie, 1904, ii, p. 716. 92 Biological Scction, American Chemical Society [Oct. fluid condition can changes in surface tension be regarded as f actors in the ameboid movement. Jensen obtained the figure of 6000 dynes per centimeter by divid- ing the weight sustained, by the circumference of the plasmodium thread. It would have been just as logical to divide the weight sus- tained by a Steel wire, by its circumference and call the quotient the surface tension of the Interface steel-air. Countless measurements of the surface tensions of aqueous Solu- tions (against air) have shown that the surface tension of water (about 70 dynes per cm.) cannot be raised very high. Some inor- ganic Salt Solutions have surface tensions against air as high as 85 dynes per cm.''' On the other band the surface tension between two Solutions such as isobutyric acid in water and water in isobutyric acid is either nil or very nearly so.^ Jensen's figure falls far outside the figures recorded for aqueous, or for any other type of, Solution, that the writer has seen. Consequently, Macallum's use of Jensen's figure and of data based upon it, in bis theory of muscle contraction based upon changes in surface tension in the working muscle, may lead to erroneous results. The elimination of zinc. William Salant and J. B. Rieger. (Pharmacological Lahoratory, Bureau of Chemistry, U. S. Dep't of Agriculture, Washington, D. C.) The experiments were made on rabbits. Zinc malate was given intravenousl}^ and zinc acetate sub- cutaneously. The urine collected for periods of 24-48 hours showed the presence of 1-2 mg. of zinc. Much larger amounts were found in the feces and contents of the posterior intestinal canal, after the subcutaneous injections. The quantities of zinc varied between 8.5 and 17.1 mg. in 24-48 hours, which represented 10-34 percent of the amounts introduced. The amounts of zinc eliminated by this Channel were greater after intravenous injection, being 17-20 mg., or 40 percent of the quantity administered. The absorption and fate of tin in the body. William Salant and L. P. Treuthardt. (Pharmacological Lahoratory, ■^Berg: Biochemical Bulletin, 1912, ii, p. loi. «Whatmough: Ztschr. f. physikal. Chem., 1901, xxxix, p. 184; Antonow: Jour. de chimie physique, 1907, v, p. 370. I9I3] I' K. Phelps 93 Bureau of Chemistry, U. S. Dep't of Agriciilture, Washington,, D. C.) Tin, in the form of a double salt, was given subcutaneously and by mouth to different animals. Analyses of the urine and feces, and of the contents of the stomach and intestines, which were made gravimetrically and volumetrically, gave the following results : After the subcutaneous administration 5-15 percent was eHminated in the urine in 24-48 hours. The feces of the corresponding period contained much smaller amounts. The contents of the stomach and intestines, and the feces, contained as much or more tin than the urine. In some animals the amount of tin eliminated by the kid- neys was smaller than that recovered from the gastro-intestinal contents and feces. Analysis of the skin indicated the presence of 20-25 percent of the amount of tin injected. When double salts of tin were given by mouth, small quantities of it were found in the tissues and in the urine, indicating that absorption from the gastro-intestinal canal takes place to a very small extent only, and may be insignificant in some animals. The amount of tin found in the liver of rabbits at the end of 48 hours varied between 0.6 and 10.8 percent. The kidneys of such animals contained quantities varying between 1.6 and 8.2 per- cent of the amount of tin injected. Experiments on the absorp- tion of the salt from the blood indicate that 85-95 percent may dis- appear in 2—3 hours af ter intravenous injection of 70-200 mg. of tin. The fate of creatine and Creatinine when administered to rabbits.^ V. C. Myers and M. S. Fine. (Laboratory of Patho- logical Chemistry, New York Post-Graduate Medical School and Hospital.) When creatine is administered subcutaneously to rabbits, in amounts varying between 50 and 100 mg. per kg. of body weight per day, 25-80 percent, depending upon the amount given, reappears in the urine unchanged, 2-10 percent is eliminated as Creatinine, and about 15 percent is retained by the muscle. If intro- duced in small amounts, as much as 50 percent may be metabolised. We are inclined to attach considerable significance to the slightly in- creased excretion of Creatinine as indicating the metabolic relation- ship between these two substances. The creatine content of the 8 Myers and Fine: Jour. Biol. Chetn., 1913, xvi, p. 169. 94 Biological Scction, American Chemical Society [Oct. nuiscle was raised from the normal of 0.52 percent to 0.55 percent (5 exp'ts) after the administration of creatine and to 0.56 percent (3 exp'ts) after the administration of Creatinine. Comparison of the observed and computed heat production of cattle.^^ H. P. Armsby. {The Institute of Animal Ntitrition of The Pennsylvania State College.) The role of oxidases in the curly-dwarf disease of potatoes. H. H. BuNZEL. {Laboratory of Plant Physiology, Bureau of Plant Industry, U. S. Dep't of Agriculture, Washington, D. C.) A method for the estimation of total fat in infants' stools. W. S. HuBBARD and D. M. Cowie. {Pharmacy and Pediatric De- partments, Uniz'crsity of Michigan.) The estimation of rafiBnose by a modified biological method. C. S. Hudson and T. S. Harding. (Division of Carbohydrate Investigations, Bureau of Chemistry, U. S. Dep't of Agriculture, Washington, D. C.) The occurrence of a toxin in the bread mould, Rhizopus ni- gricans.'^^ R. A. GoRTNER and A. F. Blakeslee. {The Carnegie Institution of Washington.) Studies in the comparative physiology of purine metabolism. Andrew Hunter, M. H. Givens, and C. M. Guion. {Depart- ment of Physiology and Biochemistry, Cornell University Medical College, Ithaca, N. Y.) The calcium content of tuberculous areas in lung tissue. Max Kahn. {Biochemical Laboratory, Columbia University.)''-^ Metabolism studies of five cases of endarteritis obliterans. Max Kahn. {Chemical Laboratory, Befh Israel Hospital, New York.y^ A dififerential stain for mucins and mucoids. Louis Berman and Wm. J. Gies. {Biochemical Laboratory, Columbia Univer- sity.)^^ A study of the influence of extemal hemorrhage on the parti- ^^ Armsby: Jour. Amer. Chem. Soc, 1913, xxxv, p. 1794. 10 Blakeslee and Gortner : Biochemical Bulletin, 1913, ii, p. 542. 11 Kahn : Ibid., 1913, ii, p. 458. 12 Kahn : Ibid., 1913, ü, P- 544- 13 Berman and Gies : Ibid., 1913, ii, p. 547. 1913] I' K. Phelps 95 tion of urinary nitrogen. Olive G. Patterson. (Biochemical Lahoratory, Colinnbia University.)^^ Further studies of edema. Tula L. Harkey. {Biochemical Lahoratory, Columbia Uniz'ersity.Y^ Biochemical studies of selenium. Victor E. Levine, {Bio- chemical Lahoratory, Coluinhia University.y^ Pigments produced from thymol by ammonium hydroxid. Benjamin Horowitz and Wm. J. Gies. {Biochemical Lahoratory, Columhia University.)'^'^ Metabolism studies of two cases of amaurotic idiocy. Max Kahn and A. Hymanson. {Chemical Lahoratory, Beth Israel Hospital, New York.y^ Further studies of the permeability of lipin-collodion mem- branes. Samuel Gitlow and Wm. J. Gies. {Biochemical Lahor- atory, Columhia University. ) ^^ The origin and significance of salivary sulfocyanate. Max Kahn and Wm. J. Gies. {Biochemical Lahoratory, Columhia University. ) Biochemical studies of dental caries. Alfred P. Lothrop and Wm. J. Gies. {Biochemical Lahoratory, Columhia University.) I. K. Phelps, Secretary Bureau of Chemistry, U. S. Department of Agriculture, Washington, D. C. 1* Patterson : Biochemical Bulletin, 1913, ii, p. 555. 15 Harkey : Ibid., 1913, ii, p. 550. 16 Levine : Ibid., 1913, ii, p. 552. " Gies : Ibid., 1912, ii, pp. 171 and 293 ; Horowitz : Ibid., 1913, ii, p. 293. 18 Hymanson : Ibid., 1913, ii, p. 457. 19 Eider: Ibid., 1913, ii, p. 549; Gitlow, Ibid. THE BIOCHEMICAL SOCIETY, ENGLAND SCIENTIFIC PROCEEDINGS Institute of Physiology, University College, London, W. C, June II, 1913, at 8.30 p. m. — C. G. L. Wolf: A note on the estimation of lactic acid. — S. Walpole: Gas electrode for general use. — /. A. Gardner and Miss C. M. Leetham: On the respiration of fresh water fish. — /. A. Gardner and Mr. Lander: On the cho- lesterol content of the tissues of cats under various dietetic condi- tions. — K. Goadby: The action of vapors, given off from paint, upon the growth of bacteria. ROTHAMSTED EXPERIMENTAL STATION, HaRPENDEN, HerTS., July 12, 1913. — W . A. Davis and A. J. Daish: (i) The quantitative estimation of maitose in plant materials; (2) A study of the forma- tion of carbohydrates in the mangold leaf. — H. E. Annett: The sugar of the Indian date palm. — A. Appleyard: Apparatus for studying the gaseous reactions in soils. — W. Buddin: Plauts grown on sterilised and unsterilised sick soils. — A. J. Prescott: A rapid method for determining P2O5 in plant ash, soil extracts, etc. — H. B. Hutchinson: Effect of lime on the biological relationships of the soil. — H. B. Hutchinson and K. McLennan: Cellulose decomposi- tion and nitrogen fixation. The beginnings of an atlas of soil bacteria. Laboratory of Pathology, St. Thomas's Hospital, S. E., Octoher 10, 1913, at 8.30 p. m. — C. Funk: The chemical composi- tion of the different parts of the maize grain obtained during the process of milling with reference to the etiology of pellagra. — C. Funk: Apparatus for concentrating Solutions of highly unstable substances. — H. MacLean: (i) A simple method for preparing pure lecithin; (2) The action of lecithin as an activator of cobra venom in haemolysis; (3) Some observations on (a) acetone soluble Phosphatides, (&) the part played by Phosphatides in fatty degenera- tion; (4) The Separation of "lecithin" into two somewhat similar 96 1913] The Biochemical Society, England 97 bodies, but containing different bases. — H. J. Page: The pigments of the brown algae. — A. C. H. Rothera: On mammary secretion. MEMBERS ELECTED June II. — E. W. H. Cruickshank, M.B. ; A. T. Daish; H. von Euler; C. G. P. Laidlaw; J. B. Leathes, F.R.S. ; V. Lefebure, B.Sc. ; V. Steele ; C. H. Warner ; George Winfield. July 12. — Alfred Barnes; Winifred Cullis, D.Sc. ; George Graham; O. C. Grüner, M.D. ; P, W. Latham; R. Stenhouse Wil- liams, M.D. Oct. 10. — G. von Anrep; U. N. Brahmachari; Mary Fräser, B.Sc; Edward Horton; Constance Leetham, B.Sc; A. B. Macal- lum, Jun., M.D. ; C. Myers Ward, M.D. ; H. S. Raper, D.Sc. ; A. Stead; Harold Wager, F.R.S. THE AGRICULTURAL COLLEGES AND EXPERI- MENT STATIONS IN THE UNITED STATES I. THE AGRICULTURAL EXPERIMENT STATIONS IN THE UNITED STATES -E. H. Jenkins.i Alabama — College Station: Auburn; J. F. Duggar.i Canebrake Station : Uniontown; L. H. Moore.i Tuskegee Station : Tuskegee; G. W. Carver.i Alaska — Sitka: C. C. Georgeson.2 Arizona — Tucson: R. H. Forbes.^ Arkansas — Fayetteville: Martin Nel- son.i California— 5^r^^/^j; T. F. Hunt.i Colorado — Fort Collüts: C. P. Gillette.^ Connecticut — • State Station : New ' Haven; Storrs Station; Storrs; Delaware — Newark: H. Hayward.^ Florida — Gainesville: P. H. Rolfs. ^ Georgia — Experiment: R. J. H. De- Loach.i Guam — Island of Guam: J. B. Thomp- son. 2 Hawaii — Federal Station: Honolulu; E. V. Wilcox.2 Sugar Planters' Station: Honolulu; H. P. Agee.i Idaho — Moscow: W. L. Carlyle.i Illinois — Urbana: E. Davenport.i Indiana — Lafayette: A. Goss.i Iowa — Arnes: C. F. Curtiss.i Kansas— Manhattan: W. M. Jardine.i KENTVCKY—Lexington: J. H. Kastle.i y W. R. Dodson.-" Louisiana — State Station : Ba- ton Rouge; Sugar Station : Au- dubon Park, New Orleans; North La. Station ; Calhoun; Rice Station: Crowley; Maine — Orono: C. D. Woods. ^ Maryland — College Park: H. J. Pat- terson.i Massachusetts— ^m/f^rj^- W. P. Brooks. 1 Michigan— £o.yf Lansing: R. S. Shaw.i Minnesota — University Farm, St. Paul: A. F. Woods.i Mississippi— /^^rncM/^wra/ College: E. R. Lloyd.i Missouri — College Station: Columbia; F. B. Mumford.i Fruit Station : Mountain Grove; Paul Evans.i Montana— 5 o^r^man.- F. B. Linfield.i Nebraska — Lincoln: E. A. Burnett.i Nevada — Reno: S. B. Doten.i New Hampshire — Durham: J. C. Kendall.i New Jersey— A''^w Brunswick: J. G. Lipman.i New Mexico— 5" tof^ College: Fabian Garcia.^ 1 Director. 2 Special agent in charge. 98 3 Acting director. I9I3] in the United States 99 J-B. W. Kilgore.i New York — State Station: Geneva; W. H. Jor- dan.^ Cornell Station: Ithaca; W. A. Stocking, jr.3 North Carolina — College Station: West Raleigh; State Station; Raleigh; North Dakota — Agricultural College: J. H. Worst.i Ohio — Wooster: C. E. Thorne.^ Oklahoma — Stülwater: L. L. Lewis.^ Oregon — Corvallis: J. Withycombe.^ Pennsylvania — State College: R. L. Watts.i State College: Institute of Animal Nutrition, H. P. Armsby.i Porto Rico — Federal Station: Mayaguez; D. W. May.2 Sugar Producers' Station : Rio Pie- dras; J. T. Crawley.i Rhode Island — Kingston: B. L. Hart- well.i South Carolina — Clemson College: J. N. Harper.i South Dakota — Brookings: J. W. Wilson.i Tennessee — Knoxville: H. A. Mor- gan.i Texas — College Station: B. Young- blood.i Utah — Logan: E. D. Ball.^ Vermont — Burlington: J. L. Hills.^ Virginia — Blacksburg : S. W. Fletcher.i Norfolk: Truck Station, T. C. John- son.i Washington — Pullman: Ira D. Car- diff.i West Virginia — Morgantown: E. D. Sanderson.i Wisconsin — Madison: H. L. Russell.^ Wyoming — Laramie: H. G. Knight.i IL THE AGRICULTURAL COLLEGES IN THE UNITED STATES Alabama — Auburn: Charles C. Thach.* Normal: W. S. Buchanan.* Tuskegee Institute : Booker T. Wash- ington.s Arizona — Tucson: Arthur H. Wilde.* Arkansas — Fayetteville: Martin Nel- son.^ California — Berkeley: T. F. Hunt.^ Colorado — Fort Collins: Charles A. Lory.* Connecticut — Storrs: C. L. Beach.* Delaware — Newark: Geo. A. Harter.* Dover: W. C. Jason.* Florida — Gainesville : J. J. Vernon.^ Tallahassee: Nathan B. Young.* Georgia — Athens: Andrew M. Soule.* Savannah: R. R. Wright.* Hawaii — Honolulu: J. S. Donaghho.'^ Idaho — Moscow: W. L. Carlyle.^ Illinois — Urbana: E. Davenport.^ Indiana — La Fayette: J. H. Skinner.^ Iowa — Arnes: R. A. Pearson.* Kansas — Manhattan: H. J. Waters.* Kentucky — Lexington: J. H. Kastle.^ Frankfort: G. P. Russell.* Louisiana — Baten Rouge: Thos. D. Boyd.* New Orleans: J. S. Clark.* Maine — Orono: R. J. Aley.* Maryland — College Park: H. J. Pat- terson.* Princess Anne: T. H. Kiah.^ Massachusetts — Amherst: Kenyon L. Butterfield.* Michigan — Bast Lansing: J. L. Sny- der.* Minnesota — University Farm, St. Paul: A. F. Woods.e Mississippi — Agricultural College: G. R. Hightower.* 1 Director. ^ Special agent in charge. ^ Acting director. * President. ^ Principal. ^ Dean. ^ Acting President. ^ Acting dean. loo Agricultural Colleges and Experiment Stations [Oct. Alcorn: J. A. Martin.* Missouri — Columbia: F. B. Mumford." Jeßerson City: B. F. Allen.* Montana — Boseman: Jas. M. Hamil- ton.* Nebraska — Lincoln: E. A. Burnett.^ Nevada — Reno: Joseph E. Stubbs.* New Hampshire — Durham: Edw. T. Fairchild.* New Jersey — New Brunswick: W. H. S. Demarest.* New Mexico — State College: G. E. Ladd.* New York— Ithaca: W. A. Stocking, jr.8 North Carolina — West Raleigh: D. H. Hill.* Greenshoro: James B. Dudley.* North Dakota — Agricultural College: J. H. Worst.* Ohio — Columbus: H. C. Price.^ Oklahoma — Stillwater: J. H. Con- nell.4 Langston: Inman E. Page.* Oregon — Corvallis: W. J. Kerr.* Pennsylvania — State College: Edwin E. Sparks.* Porto Rico — Mayagues: F. L. Stevens.^ Rhode Island — Kingston: Howard Edwards.* South Carolina — Clemson College: W. M. Riggs.* Orangeburg: R. S. Wilkinson.* South Dakota — Brookings: R. L. Slagle.* Tennessee — Knoxville: Brown Ayres.* Texas — College Station: Charles Pur- year.''^ Prairieview : E. L. Blackshear.^ Utah — Logan: J. A. Widtsoe.* Vermont — Burlington: J. L. Hills.^ Virginia — Blacksburg: J. D. Eggles- ton.* Hampton: H. B. Frissell.^ Washington — Pullman: E. A. Bryan.* West Virginia — Morgantown: E. D. Sanderson.ß Institute: Byrd Prillerman.* Wisconsin — Madison: H. L. Russell.^ Wyoming — Laramie: C. A. Duniway.* HI. ORGANIZATION OF THE OFFICE OF EXPERIMENT STATIONS, U. S. DEPARTMENT OF AGRICULTURE, WASHINGTON, D. C. A. C. True, Ph.D., D.Sc, Director; E. W. Allen, Ph.D., Assistant Director. Relations with State Agricultural Experiment Stations The Director and Assistant Director; W. H. Beal, W. H. Evans, J, I. Schulte. Experiment Station Record. E. W. Allen, Ph.D., Editor. H. L. Knight, B.S., Assistant Editor. L. W. Fetzer, Ph.D., M.D., Agricultural Chemistry, Agrotechny, and Veterinary Medicine. W. H. Beal, A.B., M.E., Meteorology, Soils, and Fertilizers. W. H. Evans, Ph.D., Agricultural Botany, Bacteriology, and Vegetable Pathology, W. E. BoYD, Ph.B., Agricultural Botany, Bacteriology, and Vegetable Pathology. J. I. Schulte, B.Agr., Field Crops. G. M. TucKER, Ph.D., Field Crops. * President. ^ Principal. ^ Dean. ^ Acting president. ^ Acting dean. I9I3] in the United States lOI E. J. Glasson, B.S.A., Horticulture and Forestry. C. F. Langworthy, Ph.D., D.Sc, Foods and Human Nutrition. H. L. Lang, B.S., Foods and Human Nutrition. H. Webster, B.S., Zootechny, Dairying, and Dairy Farming. W.A. Hooker, D. V.M., Economic Zoology, Entomology, and Veterinary Medicine, R. Trullinger, B.C.E., Rural Engineering. B. B. Hare, M.A., Rural Economics. C. H. Lane, M.A., B.S.A., Agricultural Education. M, D. Moore, A.B., Indexes. Insular Stations, W. H. Evans, Ph.D., Chief. Porto Rico Experiment Station, Mayagues. D. W. May, M.Agr., Special Agent in Charge. P. L. GiLE, A.B., Chemist. G. L. Fawcett, B.S., Plant Pathologist. C. F. KiNMAN, B.S., Horticuhurist. E. G. RiTZMAN, B.S.A., Animal Hus- bandman. H. R. VAN Zwaluwenburg, B.S., En- tomologist. T. B. McClelland, A.B., Assistant Horticuhurist. C. N. Ageton, B.S., Assistant Chemist. W. E. Hess, Plant Propagator. Alaska Experiment Stations. C. C. Georgeson, M.S., Special Agent in Charge, Sitka. G. W. Gasser, B.S., Agronomist at Rampart. J. W. Neal, Agronomist at Fairbanks. M. D. Snodgrass, B.S., Animal Hus- bandman at Kodiak. Hawaii Experiment Station, Honolulu. E. V. WiLCox, Ph.D., Special Agent in Charge. J. E. Higgins, B.A., M.S.A., Horti- cuhurist. W. P. Kelley, Ph.D., Chemist. D. T. FuLLAWAY, A.M., Entomologist, C. K. McClelland, M.S.A., Agrono- mist. William McGeorge, B.S., Assistant Chemist. Alice R. Thompson, M.A., Assistant Chemist. C. J. Hunn, B.S.A., Assistant Horti- cuhurist. Valentine S. Holt, Assistant in Hor- ticulture. C. A. Sahr, Assistant in Agronomy. Guam Experiment Station, Island of Guam. J. B. Thompson, B.S., Special Agent in Charge. Agricultural Education. C. H. Lane, M.A., B.S.A., Specialist in Agricultural Education. B. B. Hare, M.A., Assistant in Agricultural Education. E. Merritt, A.b., Assistant in Agricultural Education. John Hamilton, B.S., M.S.A., Farmers' Institute Specialist. J. M. Stedman, B.S., Assistant Farmers' Institute Specialist. Nutrition Investigations. C. F. Langworthy, Ph.D., D.Sc, Chief of Nutrition Investigations. R. D. Milner, Ph.B., Assistant in Nutrition Investigations. S. C. Clark, B.S., M.A., Assistant in Nutrition Laboratory. 102 Agricultural Colleges and Experiment Stations [Oct. Nutrition Investigations (con.) W. P. Garrety, B.S., M.A., Assistant in Nutrition Laboratory. A. D. Holmes, Ph.D., Assistant in Nutrition Laboratory. H. L. Lang, B.S., Assistant in Nutrition Laboratory. IV. OFFICERS OF THE ASSOCIATION OF AMERICAN AGRICUL- TURAL COLLEGES AND EXPERIMENT STATIONS A complete and revised list of these officers will be published in the suc- ceeding issue of the Biochemical Bulletin. A. C. BIOCHEMICAL BIBLIOGRAPHY AND INDEX 4. Third quarter, 1913 (July-September)^ WILLIAM A. PERLZWEIG (Biochemical Lahoratory of Columbia University, at the College of Physicians and Surgeons, New York) Explanation of abbreviations, arrangement, notation, etc. Bibliography. Titles of papers are freely shortened, minor words ignored, common terms con- veniently abbreviated or chemical Symbols substituted ; surnames of coUaborators are connected by hyphens ; most punctuation marks are omitted — all for the sake of condensation. Heavy faced Roman numerals indicate volumes; heavy faced Arabic numerals designate numbers and dates of issue (slanting lines separate numerals for months and days). Bibliographie itenis begin with em dashes. When two or more papers by the same author occur together, they are duly num- bered, and separated by semicolons, but follow the same em dash. Numerals preceding italicized names of authors indicate sequence in the bibliography {index numerals) ; numerals preceded by commas, at the ends of items, indicate initial pages of the corresponding papers. Index (subjects). The numerals in the index (page 109) correspond with the numbered items in the bibliography. Pages are not indicated. Numerals held in groups by hyphens are piain abbreviations in accord with the indications of the first numeral of each such series (see footnote, p. 109). Abbreviations of words in the index are similar to those in the bibliography. Each group of index references is terminated by a semicolon ; commas mark off subdivisions of a general index subject. Names of authors are not indexed. Journals included: Biochemische Zeitschrift (B.Z.), Zeitschrift für physi- ologische Chemie (Z.p.C), Journal of Biological Chemistry (J.B.C.), Biochem- ical Journal (B.J.), Biochemical Bulletin (B.B.)- Practical use of the bibliography. The bibliography is helpful from sev- eral Standpoints. Thus, if it is desired to ascertain whether the Journals included in the bibliography contain any papers (during the given quarter) on a particular subject, e. g., lipins, find the key word in its alphabetical place in the index and turn to the items in the bibliographic sequence indicated by the index numerals (in this case 41, 51, 72, 94, 99, 237, 416). The abbreviated items thus identified give the names of authors and suggest the nature of the corresponding papers (four papers, in the case selected for Illustration), and help the reader to decide whether to examine the original publications. When the index gives a negative answer to an inquiry, a large mass of literature is removed from further consid- eration. During the intervals between publication of the Indexes of Journals, 1 The preceding portions of this bibliography and index were published at pages 298, 470 and 559 of volume II of the Biochem. Bull. (1913)- 103 104 Biochcmical Bihliography and Index [Oct. Cetttralblätter and year books, this runningbibliography directs the readertomost of the main tracks through current literature on the leading biochemical subjects. Bibliography. B.Z. — LH : 5-6; 7/9. — lEUas-KolbSiu i Koh'hyd'- st'wechs,33i. — 2Lhotäk von LhotaVerteW u Aussch subcu Digitox, Btifo vnlg,2ß2. — 2>^Ianabe-MatulaV\\ys\k2^. Zustandsänd Kord,369. — 4Hämäläine7iSynth Glucosid d Terpenalkoh m Emulsin,409. — ^Bach Redukt'fermen,4i2; öOxydat Bild HNOg Pflanz'ej^tr,4i8. — yPugliese Physiol Milz,423. — SThar-BencslawskiZusam d nach Zn-verfahr ber- gest sog kol'd N Harn,435. — gErlenmeyerOpt-akt Verbind leb Zell; künst Darst ohn Anwend asym Molekül o asym Kräft,439. — loLesser Wirk diastat Ferm auf Glykog Zell,47i. — iii^/wyz/^rAssim'bark Maltos dur Hefen,486. — i2Neuberg-SteenbockB\\d höh Alkohol a Aldehyd dur Hefe, Valerald zu Amylalk,494. (Pp. 175.) B.Z. — LIII: 1-2; 7/15. — i^Bdron-PöldnyiZweit Hauptsatz Ther- modyn auf Vorgang Org's,i. — i4TaM(7/Calorim kl Tier,2i ; i5Calorim Nier'arb,36. — i60serna-Kelemen Arh kr Nieral. — iyVerzärMi\z3.rh, 69. — iSHannemamiR'mü Grosshirn a Stoff- u Energ'ums,8o. — igAlex- ander-ScernoEmü Narkos a Gaswechs Gehirn, 100. — 2oHdnWirk Koh'hyd a Energ'ums,ii6. — 2iVerzär-von FejerVerhr Traub'zuck Pankr'diab,i40. — 22von Fejer'E'mü Schmelzp't nich emul Fet Geschw Entleer Magen, 168. 3; 7/18. — 27,Roho7iyiKoVchem Eiw'stud,i79. — 24RohonyiRmgfig gefror Gelat,2io. — 25B^rc^£'//^rStalagmom Stud kol'd u kryst Lös,2i5; 26Ibid.,2^2. — 2'/Berc3eller-CsäkiIbid.,2^S. — 28- GroJiWirk Fe-geh Blutmehl auf Fe-ums B'mehl gefüt Tier,256. — 29- SieburgY erhal /'-Cl-m-Kresot'säu,259. 4-5; 7/21. — T,oSchIossmann- Murschhanser'E'mü vorangegang Ernähr St'wechs Hung,265. — 31- Scheunert-Grimmer-AndryewskyTopogra.ph Peroxydas Verd'schlauch, ihr Nachw,300. — T,2MichaeIis-PechsteinKa.ta.\3.s, Leber,320. — ;^T,Ehren- ^^r^rGelat'quel was Lös, 356. — 34Lo^^Anpas Fundulus höh Konz,39i. — 35A'^^M&^^5'-i^^^&Zuck'fr Hefegär,4o6. — 365^r^o/mJErwid Salkowski Mitt, Wirk Antisep auf Tox,420. — 375'a/^owj^iBemerk Erwid Ber- tolini,422. 6; 7/24. — ^SHämäläinenSynth /S-Glucosid d Terpenalkoh, 423. — 39Gnmm^rFermen Milchdrüs Milch,429. — 4oGaleottiKondens Am'säu verm Formal,474. — 4iStiiber'B\uÜ{p Phagocyt, 493. — 42Sonn- tagMeth Bertrand Zuck-best,50i. (Pp. 505.) B.Z. — LIV: 1-2; 7/31. — 435a^a^zPhos'tid Placen,i ; 44P-verteil Placen,5. — 4sDreyer-WalkerTheor Wassermann Reak,ii. — 46Lazvroiv Beeinfl Wirk Medik durch Lecith,i6. — 4yKopac2eiuskiD\d.\ys f anal Zweck,27. — 48Baumann'N- Bestand Kephalin,30. — 4gBentozullypo- ph'enzym,40. — soRogee-Fritsch'Nen Makr u Mik'meth quan Best Gl I9I3] William A. Perlzweig 105 Blut,53. — 5ii?w/i/aMC?Lip'd Ult'filt'theor Plas'haut, u Bedeut elek Lad Kol'd Vitalau fnahm,59. — ^2Niegovan'En\h:i.\ Milch Phos'tid, 78.-53- RifätzvachdaniSchick Cocain, Ekgonin Org's,83-— 54-^''^^^^^"^Resp'ap m selb'kont o best, verw kl Tier,92. — ^sMeisenheimer-St. Gamharjan- SemperRein Invertas Behand m Sau, 108; 56Anreich Invertas geh leb Hefe,i22. — c,ySpiroF'il KoVd,iS5- — sSGr^-^nwaWBemerk Mitt Paladino Verän Stoff wechs Tier nach Exst Schilddr u Parath,iS9. 3-4; 8/9. — ^gKatifman-Asser Aussch Morphin Harn,i6i. — 6oFriedenthalKup'\\ing Eiw'spal'prod kol'd Koh'hyd'ket,i74. — GiPorgesBezieh COg-span Blut, Lung'vent,i82. — 625'^mow^rAussch Ameis'sä Urin, 189. — 6^Lif schütz Quan Best Choles'est,2i2. — 64L^j'^^rBeeinfl Glykog'schw auton Org Frosch d Anoxybios,236; ösFehl'quel Blutzuck-best Frosch- Schild- kröt'blut,252. — 66Ehrlich-LangelJmw Asparag b Koch was Lös,256. — GySchewketOxyd Gall'säu, Gall'gerbsäu (Tann) Luft, Gegenw Alka!, Farb'reak Blei,277 ; 68Neu Farb'reak Di-, Tri-phenol,282 ; 69Farb'reak Erdalkal m Oxygarderiv,285. — yoSchreiber-LeiiardKsimoVhem Eig'sch Cholest, Oxychol,29i. — yiScaffidiLös'k Harnsäu i Essigsäu,297. — 72- Traw&^Theor Haftdru u Lip'dtheor,305 ; 73Narkos,3i6. — y ^Michaelis- David sohnWwk H'-konz a Kordgemisch,323. — y^Stoklasa-Sebor- ZdobnickyFhotochem Synth Koh'hyd,330. 5-6; 9/2. — y6König-Gross- /^/cfFischsperm als Nahr'mit f Mensch,333. — 77Fischrog als Nahr'mit f Mensch,35i. — 78M^mM^rBeeinfl Morph'wirk dur Nebenalk'd Opium, 395. — 790/ifaEig'sch Kanin'ser n Vorbeh mit Emulsin,43o; SoAbbau Harnsäu m HoOg u Fe-salz,439. — 8iM^jj'^r/iResorp'geschw Eiweis ihr Abb'prod Dünnd,446. — S2F reise COa-bild i Leber,474. — S^Grimmer Druckf'ber z Arb Perm, Milchdr, Milch,503. (Pp. 505.) B.Z. — LV: 1-2; 9/12. — 84MoyerBrenztraub'säu i Tierkörp,i. — 85- Scheu'ketNzchw Glucur'säu diab Harn,4. — SSW ohlgemuth-RewaldV trh I-eiweisOrg's,7. — 87^j/z^rDrüsen,i3. — 88Maj'^/ozc'Pf wachs Org's,45. — SgZaleski-Schataloff'E'iw'svimw Hefe,63. — 9oZa/^i-^i-5"/ia^^mEiw'saufb Pfl'z,72. — gi Karczag-MöczärVergär Brenztraub'säu Bakt,79. — g2Fasal Tryptophangeh Hautgeb u malig Tumor,88. — 93i^aurf^rj-Choles'estergeh Blut vers Tier,96. — 94i?Mw/'/Einfl Lip'd Gerin Blut, loi. — gsDiakow Meth calor u El'anal Hilf calor Bomb,ii6. — g6DienesSt'wedis Schwang'sch, Lact, 124. — gyOppenheimerF'ix Digital'kör tier Org's, Verh z Blut,i34.— 98£/ta^C02-bild üb'leb blutdurchström Musk,i53. 3-4; 9/18. — ggKirscheL'ip'd Org'hämolys, Beeinfl dur Traub'zuck'füt, 169. — looGt^forn-Maj^miMuskulat u Clykolys,i89. — loivon der Heide- Kleinst- u Energ'ums Schwein b Wachs u Mast,i95.— I02Low/^o/^ ChristiansenBest kl Meng Hg org Subst,2i6. — 1035'a.r/Stör Eiw'st'- io6 Biochemical Bihliography and Index [Oct. wechs Krebskr; Rhod'aussch, 224. — lO^BywatersKssxm i Ei enth Eiweis d Hühn'embry,245. — losSalkowskiViX Purinb dur Zn-salz a Fleischext u Harn,254. — 106IV olterOaem Krebstum,26o. — loyThorsch Einw Alkoh auf antig rot Blutkör,266. — io8i?^wa'/N-Best Kephalin, 296. — logEmhden-BaldesAhh^in Phenylalan tier Org's,30i. — iioGries- bach-0 ppeJiheimerMilchs'iu Blut,323. — 1 1 lEmbden-OppenheimerYerh. Brenztraub'säu Tierkörp,335. — ii2ZiintzEmi chron Unt'nähr St'wechs, 341. — ii3L^jj^rBeeinf endozel Wirks Leberdiast dur Pankr'exst,355. 5-6 ; 9/25. — I i4Halle-Loewenstein-Pr{bramF3.ThTeakt Triketohydrinden hydrat (Ninhydrin),357. — iic^SchirokichBedent Pentos Energ'quel tier Org's,37o. — I i6Fa^a/Pigm,393. — i lyCostantinoFormol titr Am'säu-N Blutk Ser Blut hung u ernähr Tier,402; iiSPermeab Blutk Am'säu, 411; iigMeth Extr Am'säu versch Best'd Blut,4i9. — i2oFriedmann- TürkAhh CO2 Tier,424; i2ilbid.,432. — i22Friedmannlhid.,4:^6. — 123- Mochisukilhid.,443- — I24lbid.,446. — i2^Friedmann-Maaselhid.,4S0. — i26von La germar kYerhr Ketoreduktas Geweb,458. — i2yFriedman- Türk Ahh Naphthalinkern Tier,463. — i28rMr^|ö-Naph'alaninhydantoin- säu,477. — i2gKotschneffRol Perm tier Org's, Einfüh getöt Tub'k'baz, 481. — i3oA^^M&^rpr-0^r?^/Meth'glyox'bild,495. — 131-200 blank. (Pp. 505.) Z.p.C— LXXXVI : I ; 7/i.—20iTrierMeth Lecith'dars Pf'z'sam erhäl Verb,i. — 202PanserBioch. Protozoen,33. — 202StanfordCereh'sp'- flüs Geist'kr,43. — 204K«jf^fHämat, Häm'porph'bild,5i. — 20Zvan Dam Bemerk. Arb Rakoczys Peps-Chymos'frag,77. 2; 7/10. — 2o6Zeller Essb indis Schwalb'nest,85. — 2oyBudaiMeth quant Best NH3, Tri- meth'am,io7. — 2oSEider-CasselA\koh. Gär,i22. — 209Z)o/jrwNucleinst'- wechs,i30. — 2ioBugHa-CostantinoMusk'ch, Wärm'trock Musk'l See- tier,i37. — 2iiTrierMeth Lecith'dars Pf'z'sam erhal Verb: Hydrol Eilecith,i4i ; 2i2lbid. ; Hafersam,i53. — 2i2YoshimuraOrg Bas getrock Rogen Hering, 174. — 2i4Yoshimura-KanaiN-ha.\tig Bestand Pilz Cor- tinel shiitak P. Henn,iyS. 3; 7/15. — 2ie,Küster'H.Sima.t: Meth'ier Hämin, Br an Dimeth-(Cl)häm, Dimeth'-(Br)häm,i85. — 2i6Brabant Homol Muscarin i C3-Reih,2o6. — 2iyBallowitsVork alkoh'beständ karminr u braunr Färbst Haut v Knoch'fisch,2i5. — 2i8Stanford Cereb'sp'flüs Geist'kr: Quan Best kl Meng N,2i9. — 2igWinterstein- i^^Mf^rHistid'betain Steinpilz,234. — 220S chade-B od enAntw Bemerk Lichtwitz betref Abhand: Anom Harnsäu'lös'k (kol'id Harnsäu),238. — 22iBer Arb Hermann Abb /?-Ketonsäu,244. 4; 7/21. — 222Stieger Verbr Asparag, Glutam, Argin, Allantoin i Pf'z,245. — 223Hemiceros Wurz'stöck, Rhizom, Wurz'knol,27o. — 224Gra/^-f^m^5:N-st'wechs Füt 1913] William A. Perlzweig 107 NaN03,283. — 22^GolodetsD\2i\ys quan Best,3i5. — 226PanserYiC\ gas auf Erhitz'veränd Diastas,322. — 22yHenze'Q\vit Ascid,34o; 228Frei H2SO4 Mantel Ascidia mentula,2,AS- 5 5 7/29- — 229Gra/^N-reten bei Füt Harnst,347. — 23oi^orö.y3'Zuck'resorp,356; 23iChrphyrassim,368 ; 232Mikr'kalorim z Best Warm'prod Bakt,383. — 233Pa«^^rNH3 auf durch Erhitz unwirk Diastas,40i. — 2T,4TrierMeth. Lecith'dars Pf'z'sam erhäl Verbind Erbs, Schwarzkief, Reis,407. 6; 8/7. — 2^^Riesser Kreatinbild tier Org's : aus Betain u ChoHn,4i5. — 2^6Abderhalden- Froehlich-FuchsSpalt d//-Am'capr'säu (Norleuc) i op'ak Kompon mit Formylverb: Polypept, der Aufb Amin'capr'säu beteil,454. — 2T,yMüller- ReinbachMa.sk Blutfet Blutlip: Verdau'Hpäm b Mensch,469. — 238- HirschbergQuan Best gering Meng Traub'zuck Harn Bertrand Meth, 484. — 2T,9Philipp'N enteiw'st Blutser,494. — 24oSiebiirg'H.yd'ceph'üü.s, 503. — 24iGr^^MwaW-/aw«^3;Ameis'säu'aussch b Krank,5ii. (Pp. 512.) Z.p.C.—LXXX VII: i; 8/16.— 242Gw/ewi/^c/jExtr'st Musk'l: Car- nosin u Carnos-NOg,!. — 24T)Sm.orodin2ewIbid.: Carnosin, Meth'guan, Carnirin Pferd'fl,i2. — 244Ber Smorodinzew,20. — 2455^^^rVert Krea- tin Säugetier,2i. — 24.6F ischer-Röse A\k'la.t auf Hämin u Deriv: Aufsp Häm dur K-alk'lat, neu Bild'weis Mesoporph,38. — 247/f^w£:^p-Oxy- phen'äth'am Speich'dr'gift Ceph'pod,5i. — 2486'c/i^wc^Cholsäu,59. — 249- Lock-ThomasGeh Blutplas'prot bas Bestand,74. — 2^oThunbergBemeTk Mitt V Warburg-Meyerhof: katal Beschleung O-aufnahm Lecith dur Fe salz,82. — 2^iWarburgAntw Bemerk Thumberg,83. 2; 8/26. — 252- TamuraChem Bakt,85. — 2S2,PanserllCl u NH3 gas auf Erhitz'verän Diastas,! 15. — 2S4Hirsch-ReinbachFesse\'hyp'g[yk2Lm Fessel'glykosu Ka- nin,i22. — 255£w/ß'rKatal alkoh Gär,i42. — 256£m&^c^Bernst'säu i Flei- schext u frisch Frch,i45. 3 5 Q/h- — 2575'fan/orc?Verdün'korim, nebst Bemerk Vers' f eh kol'im Vergl,i59. — 2c^SSchumm'Na.chw Hämat mensch Blutser,i7i. — 25950/wEnzymgeh Blät Salix caprea,iS2. — 26oStanford Indigobild Subst Harn (Harnindikan),i88. — 26i6'f^M Forma indophenol nucrrplasm membr frog bl'd corp, accel b ind shock,237. — 426Thom-Cnrne'Roq'i'rt mold chees,249. — 427 Gor eVolsitil HjSO^ i vacu dr'ng,259. — 428I>oHn-Z)M(//^3'Racem'- tion prot'ns, deriv fr tautom chang, Racem casein,263 ; 429Enz on racem prot, fate anim body,27i. — 4;^oKendall-WalkerBa.ct metab: Deter Urea N cultur bact,277. — 42,iMyers-FineStarv upon creatin cont muscl, 283 ; 432Carb'hyd feed creatin cont muscl, 305. — 42T)Osborne-Mendel- Ferry-WakcmanGTowih. sl ch constit diet,3ii. — 4^4U nderhillM ttah NH^-salt: Elim inges NH4-salt,327 ; 435lbid. : Elim ingest NH^-salt, prolong inanit,337. — 426Underhill-Goldschmidtlhld. : Util NH^ salt non-N diet,34i. — 42yAbderhaldenRem com Folin-Denis,357. — 438- L^z/^i^Cer'br'sid brain,359. — 4^gMurlin-Kramer'P^ncr a duod extr on glycosur, resp metab depancr dog,365. 9; 3. — 440W oodrujf-Underhill Protoz prot'pl indicat pathol chang: Nephritis,385. — ^441 Ibid. : Car- cinoma,40i. — 442Emerson-Cady-Bailey'ilCN fr prot,4i5. — 443C/aw- son-YoungFrod HCN bact,4i9. — 444Koch-KochChem dif centr nerv syst : brain alb rat, growth,423. — 445LoMöfAdenas hum body,449. — 446- r>a^m-I?Mcf/^yGlyoxalas : Distr, a rel to pancr,463. — 44yLevene-Meyer 1913] William A. Perhweig 109 Leucocy a tis on c?/-alan,475. — ^4^Levene-LaF orgeCd^st pentosur,48i. — 449Marj-/ia//Meth deter urea i hV6.A^7- — 45oDeterm urea i urin,495. — 4^iMacleodWCd glycolys : Carb'h'dr metab, "Sucre virt" i fr bl'd, 497. — 4S2johns-BanmannFur'm : 2-ox-6-meth-9-eth'p, 2-oxy-6, 8-di- meth-9-eth'p, 2-ox-6-meth-8-thi-9-eth'p, 2-ox-6-meth-9-eth-pur-8-thio- glycol-ac, 2-meth'mercap-6-ox-8-thiopur,5i5. — 453-600, blank. (Pp. 528.) BJ. — VII: 7/4. — 6oiNevilleFat y'st,34i. — 6o2EvansC3.rhona.t Ce, La, Y grow a cell-div hyacinth,349. — 6oT,Fiink-MacallumSuhst fr alcoh ext o foodst giv col reac w phos'tung- a phospliomolyb-ac,356. — 604- GrcyTrod acetald dur anaer ferm glucos b B col com,T,S9- — 605- Smedley-LiibrzynskdB'ioch synth fat-ac,364. — 6o6Conden arom aldeh w pyruv-ac,375. — ßoyChick-MartinFrecip egg-alb amm sulf : " Salt-out " prot,38o. — 6o8Mi7ro3;Est urea,399. — SoglValpoleGsLsd&ctrod gen use, 410. — 6ioStephensonEster palm-ac,429. — 611-700, blank. (Pp. 95.) B.B. — II: 8; 7. — 70iCro//Modiff Meigs meth quant deter fat milk, w impr app,509. — yo2GreavesAs i soils,5i9. — yo^Harris-Gortner'Rel w't sug-beet a comp juic,524. — 704//armBarom pres a CO2 excr man, 530. — 705Gor^w^rBleaoh flour decis,532. — 7065'öV^M.y^wHansen Fund, 535. — yoyGibsonBiochem i Philippin, 536. — yoSP .H .D .Fh.D . in biochem Amer Univ, 1912-13,538. — yogLothropFroc Col Univ Bioch Assoc, 541. — 7ioG/^i'Bioch bibl index,559. — /iiBioch news, notes, comment, 567. — 7i2Editor (Mathews plan Amer Biol See), 582. (Pp. 96.) Subject index. Absorp8i,230-S(>-7i-5,409;2 ac'aldeh6o4; acet-ac7i ; acidi,55; adenas445; age402; alan4i2-47; alcohi2,iO7,2o8-5S,422,ates246,extr603; aldehi2, 606; alkali67,earth69; alkard79,97; allantoin222 ; Amer-Biol-Soc.712; am-ac40, 117-^-9.41 i>c?^-am-capr-ac,236; NH3207-33-53 ; NH4salt434-5-ö,sulfat6o7 ; am'alcoh 12; anoxybios64; antidot4i5; antigeni07; antisep36; appari4-5,47,54,95,232-S7, 701; argin222; arrowhead423 ; AS702; ascidian227-5 ; asparag66,222 ; assimili04, 231 ; autodiges4o6. B.-co/-com6o4; bact9i,i29,232-52-62,443,6o4,metab43o; barpress 704; base2i3,403; beet703; Bertrand-meth42,238; betain23S,histidin2i9; bibliog- bioch7io; bile267; Biochem: news-note-com7ii ; Ph.D7o8; Philipp707; bird-nest 206 ; bleach-fl'r704 ; bl'd4i,5o,6i-5,93-^-7,iio-9,227-37-49-67-76,449-5i,corpi07-i7-^, 425,dry28,lipin(oid)4i,237,ser79; boletus-yel2i9 ; brain 18-9,438-44; Br2i5; Bufo- vulg2. CaIorimi4-5,95,232 ; canci03-ö; carbohydi,20,6o,75,432,met45i ; CO3602; €0261,82,98,120-1-^-^-^-5,704; carcinom44i ; carnirin243; carnosin242-j;casem 424-8; castrat4i8; catalas32; catalys25o-5 ; cell9, 10,263-5,602; ceph'pod247; cere- bron-ac4i9; cereb'sid438; cereb'sp-fl203-i8; cerebrumi8; Ce6o2; chees426; CI50; /'-Cl-OT-creosot-ac29 ; chrphyl23i ; cholest63,70,268,ester93 ; chol-acid248 ; cholin 2 This series of abbreviations, illustrating all others in the index, represents the following sequence of numerals : 230, 250, 271, 275, 409. The numerals in bold-face type here are omitted from the abbreviations above. 1 10 Biochcmical Bibliography and Index [Oct. 235; chondr-H;S04405-i4 ; chymos205; circ4o8; cleav-prod6o,8i ; clot94; cocain 53; cord3,23-5-<5-7,5 1-7,60,74,220, N8; colorim257; col-reac67-5-9,ii4; Col-Univ- Bioch-Assoc709 ; concent34; condens40,6o6 ; correc'n83; ccrrelat703; Cortinel- edodes273, shiitak2i4; cot-seedmeal4i5; creatin235-44,43i-^; cryst'oid27; cycla- min268; cholest268. Depres-subs42i ; desic2io,427; diabet2i,85; dialys^r(is)47, 225; diast 10, II 3,226-33-53; diet402-i6-5-33-<5 ; diges237-7i,trac3i ; digital97; digi- tox2; diseasi6,203-i8-4i-7i ; distrib44,i26,222,244,446; doctorate7o8 ; drug46; duod- extr439. Eggio4,2ii-66,alb6o7,yolk266; ekgonin53; elecS 1,425,609; embryoi04; emulsin4,79; energloi-i5,exchangi8,20,ioi ; enzym4,S,io,3i-i'-9,49,55-ö,79,83,ii3- 26-9,259-63-^-5-91,429-46; epineph42o; ester6io; excr59,62,i 03,241. 704; extract(iv) 6,242-5-56,439. Fast30,ii7; fat22,409,6oi,70i ; f at-Iike-sub409 ; fattenioi; fat-ac 605; ferm't'n35,9i,2o8-55,6o4; fish2i7,roe77,sperm76; fixat97; food76-7, 112,206, 401,603; formaId40,ii7; form-ac62,24i ; form-comp236; frog-urin272; fund.Han- sen7o6; Fundulus34; fungi2i4-9. Gall-ac67; gas-electr6o9; gelatin24,33 ; gland 39,87; gluc'ogen4i2 ; d-glucosam262 ; glucos2i,99,238,4i7,6o4,id4,38; glucosur254, 439; glucuron-ac85 ; glutam222; glycogio,64; glycolysioo,45i ; glyoxali30,ase446; grow88,ioi,4i6-33-44,6o2. Hansen-fund7o6; heat-prod232 ; hematin204-iS-58; hematoporph204 ; hemicerios223 ; hemin2i5-46-76; hemoglob276; hemolysm(is) 70,99,268; herring2i3; hexos404; histidin-betain2i9; hyacinth6o2; hydroceph-fl 240; HCI226-53; HCN442-J; H*conc74; H2O280; hydrol2ii,4i3; 0H-ac4ii ; hypophj'S49; hyp'glycem254. Inanit435 ; index-bioch7io; indican26o-9 ; indicat 440-7; indigo26o; indophenol425 ; inter-metab4i2; invertas55-ö; iodid 86; I-prot 86; Fe28,8o,250,4i5,salt250. Kephal48,io8; /3-ket-ac22i ; a-ket-ald4ii ; ketoreduc 126; kidni5-6. Lactat96; lac-aciio,274; La6oi ; Pb67; Ieaf259; lecith46,20i-ii- ^-34; leucin236; Ieucocy447; lignocer-ac4i3-9 ; lipem237; lipm(oid)4i, 51, 72,94-9, 237,416; liv32,82, 113,408; lung6i. Maltosii ; mam-gl39,83; Mathews-pl7i2; meal 415; meat256,40i,extrio5; Meigs-meth70i ; melt-p't22; membr425; Hgi02; meso- porph246; metabi,i8,28,30,58,96,ioi-j-i2,209-24,40i-5-i2-30-<^-5-ö-9-5i ; meth4, 12-^-5, 42-7,50-^-5,63-5.85,95, 102-19-30,201-7-1 1-^^-25-32-.^^, 63-5-74, 407-20-30- 49-50,608, 701; meth'at'n2i5; meth'glyoxi3o; meth'guan243 ; microcarim232; mik'meth50; milk52,83,7oi ; moId426; morphin59,78; muscar2i6; musc98,ioo,2ic>- 42-j,422-3i-^,extr242. /3-Naph'alan-hydant-aci28; naphthal-nucl27 ; narcoi9,73; nephrit44o; nerv : dis203-i8,syst444; nest(bird)2o6; ninhydrii4; NOs224; HNO, 6; N8,48,io8-i7,2i4-5-24-9-39,4o8-30.retent229; non-N-diet436; norleucin236; nucleas269; nucl-ac273-5 ; nucl-metab209; nucrhist26i. Oat2i2; opium78; opt- act-conip9; oxidat6,67; oxycholes70; oxygallol69; 054,250; /'-oxyphen'eth'am 247. Palm-ac6io; pancrii3,446,diabet2i,extirpii3,extr439,secr40i ; parathyr58,-ect 58,403; pathol62,92,440-7 ; pea234; pentosii5; pentosur448; peps205; permeabii8; peroxidas3i; phagocyt4i ; phenol68; phen'alani09; Philippin707; phos'tid43,52, 266; phos'molyb-ac6o3 ; P44,88; phos'tung-ac6o3 ; pho'syn75; pigmi 16,217-67-76; placen43-^; plant6,90,222 ; plasma5i,249,membr5 1,425; polem36-7,58,205-20-5o-/, 437; polypep236; porphyr276; precip'n57,io5,6o7; pregn96; proceed'g709; protec- enzym264; prot'n23,6o,8i-6-9,9O,i03-4,249-6i,402-28-9-42,6o7,iod86 ; protopl440-J ; protoz202,440-J ; purin4io, (oxy,meth,eth,thio,glycol,mercap)452 ; pyrotar-ac84,9i, III ; pyruv-ac4i2-7,6o6. Racemiz'n428 ; racem-prot429 ; reductas5,i26; renn424; respi9,54,6i,704; resp-metab439; respirim54; retent229; rhizom223; rice234; roe 77; root223; Roquef-niold426. Saliv-gl247; Salix-capr25g; salt-out6o7; secr40i ; seed20i-ii-i'-34,4i5; ser79,ii7,239-58,prot'n402; skin2i7; sm-intes8i,275; NaNOs 224; soil702; specifty263-^-5; sperm76,2i3; sphin'myel4i3 ; sprn7,i7; stalagmom 1913] William A. Perlsweig in 25-Ö-7; starv43i; stom22; succin-ac256; " suc-virt "451 ; sugar42,65,23o,423-si, beet,703; sulfocyani03; sulf-ac228,427; supra-gl420; swall-nest2o6; swell33; synth 4,90,605. Tann67; tautom-chang428 ; terpen-alc4,38 ; test4S,69, 114,270; ther'dyn 13; thymu 558,406; tiss92, 126-37,274,404-21-38-47; toxtc(in)36,403,4i5; trik'hydr'- hydii4; trimeth'am207 ; tryptoph92; tub'c-bacili29; tumor92,io6,4i8; tyros407. Ultrafilsi; und'feedii2; urea229,43a-49-5o,6o8 ; ur-ac7i, 80,220; urin8,59,62,85,io5, 238-60-72,403-21-50. Vac-dry427; valeraldi2; volat427. Wassermann-test45. Y'stii-^,35,s6,89,275,6oi,nucl-ac273; yolk266; Y602. Zn-saltsi05. BIOCHEMICAL NEWS, NOTES AND COMMENT EDITORIAL SUB-COMMITTEE: Alfred P. Lothrop, Walter H. Eddy, Arthur Knudson, Paul E. Howe, Emily C. Seaman. Contents.— I. General: Necrology, 112; honors, 112; resignations, declina- tions and appointments, 113; Alvarenga prize, 115; grants, 116; endowment funds, 116; notes on radium, 117; meetings of societies and congresses, 122; miscellaneous items, 126. IL Columbia Univ. Biochem. Assoc.: (i) General notes, 129; (2) proceedings, 131; (3) Columbia Biochem. Dep't, 131. I. GENERAL Necrology. John G. Curtis, emer. prof. of physiology, Colum- bia Univ. — /. Lucas-Championniere, who introduced antisepsis into France. — Jules Ogier, formerly president of the Societe de chimie, a member of the Comite consultatif d'hygiene publique de France, author of Traite de chimie toxicologique. — Friedrich Seiler, prof. of pharmaceutical chemistry, Lausanne. — Charles Tellier, the in- ventor of the cold storage System. — William T. Wenzell, emer. prof. of chemistry, Univ. of Cal., Col. of Pharmacy. Honors. Honorary degrees. Dr. Svante Arrhenius (Nobel Institute, Stockholm) and Madam Curie (Sorbonne, Paris) were among the foreign representatives at the recent meeting of the British Assoc. for the Adv. of Science upon whom the Univ. of Birmingham conferred its doctorate of laws. (See p. 117). Presidency of the British Assoc. for the Adv. of Science. Prof. William Bateson, director of the John Innes Horticultural Institute, has been elected president of the British Assoc. for the Adv. of Science for the meeting to be held next year in Austraha. Ovation. Prof. /. L. Prevost of Geneva was given an ovation recently on the occasion of his reaching the time limit of age and giving up the chair of physiology which he has so long filled. Dele- gates from medical societies and universities were present from 112 1913] General 113 France, Italy and other countries, and several decorations and de- grees were conferred on him. He was one of the founders of the Revue Med. de la Suisse Romande, now in its thirty-third year, and has been a member of the editorial staff from the beginning. Legion of honor. Dr. Roux, director of the Pasteur Inst., has been made a grand officer of the Legion of Honor. AwARDS OF PRiZES. The Academie des sciences has awarded a Montyon prize ($500) to Dr. L. Amhard for his memoir on the Renal secretion. — The Raymond Horton-Smith prize at the Univ. of Cambridge for 1913 has been awarded to F. A. Roper and F. S. Scales, who are adjudged equal for theses for the degree of M.D. Their subjects were : Creatinin and creatin metaboHsm, especially in reference to diabetes, and The electrocardiogram in diabetes. — The Warren triennial prize for 19 13, amounting to $500, has been awarded to Dr. Prof. Arrigo Viseritini, instr. in pathol. anatomy in the Royal Univ. of Pavia, for his essay on the Function of the pan- creas and its relation to the pathogenesis of diabetes. — The Intern. Med. Congr., at its London meeting, awarded its three prizes as fol- lows : The Moscow prize to Prof. Charles Richet (Paris), for his work on anaphylaxis; the Paris prize to Prof. A. von Wassermann (Kaiser Wilhelm Inst, for Exp. Therapy), for his work on exp. therapy and immunity; and the Hungary prize to Sir Almroth Wright (London), for his work on anaphylaxis. AwARD OF MEDAL. The Baly medal has been awarded, by the Royal Col. of Physicians, to Dr. /. S. Haidane, F.R.S., reader in physiology at the Univ. of Oxford. Resignations, declinations and appointments.^ Declina- TiONS. Professor His, who was asked to accept the appointment of director of the med. clinic as successor of von Noorden, at Vienna, has declined the honor. — Dr. Franz Knoop, assoc. pro f. of physiol. chemistry, at Freiburg, has declined appointment to membership in the Rocke feller Inst. Appointments. Ala. Polytech. Inst, and Exp. Station: Dr. L. S. Blake, head of the dep't of pharmacy, vice Prof. E. R. Miller resigned. ^ In this summary institutions from which resignations occurred are named in parenthesis. See page 129. 114 Biochcmical Nczvs, Notes and Commenf [Oct. Bryn Mawr College: Dr. A. R. Moore (Univ. of Cal.), prof. of physiolog}'. Carnegie Institution, Nutrition Lab. (Boston): Dr. Carl Tigerstedt (Physiol. Inst., Univ. of Helsingfors), research associate. Clark Univ. : Prof. R. S. Lillic (Univ. of Penn.), head of the biolog. dep't. Cornell Univ. ]\Ied. Seh.: Dr. Joseph C. Bock (Carnegie Nutrit. Lab., Boston), instr. in chemistry. Inst, for Infec. Diseases (Berlin) : Prof. Friedrich Loeffler (Greifs- wald), director, succeeding Prof. Gaffky. Iowa Agric. Exp. Sta. (Arnes) : Dr. P. L. Blumenthal, assis. chemist, to conduct investigations on the chemistry of orchard insecti- cides and fungicides. Jefferson Med. Col. : Messrs. C. A. Smith (Penn. State Col.), demonstrator and Maxzvell Silhnan, instr., in physiol. chemistry. Kaiser Wilhelm Inst, for Exp. Therapeutics (Berlin) : Prof. A. von Wassermann, director; Prof. Carl Neuberg, demonstr. in the chemical division. Kaiser Wilhelm Society, Research Inst, for Biology: Dr. Carl Correns (prof. of botany, Münster), director; Dr. Hans Spemann (prof. of zoology, Rostock), assis. director; Dr. Warburg, in charge of the work in cell physiology. Mass. Inst. Tech.: Mr. R. S. Weston, assis. prof., dep't of biology and public health. Peter Bent Brigham Hosp. (Boston) : Dr. Francis H. McCrudden (Rockef eller Inst.), director of the laboratories. St. Louis Univ. Seh. of Med.: Dr. Don R. Joseph (Bryn Mawr), prof. of physiology. U. S. Dep't of Agric, Bur. of Chem. : Dr. Isaac K. Phelps (Bur. of Mines, Pittsburgh), chemist; chief of the division of organic chemistry investigations. U. S. Public Health Service, Hygienic Lab. (Wash.) : Dr. Carl Voegtlin (Johns Hopkins Univ.), prof. of pharmacology ; Dr. E. B. Phelps (Mass. Inst, of Tech.), prof. of chemistry. Univ. of Ala. Med. Seh. ; Dr. Andrew H. Ryan (instr. in physiology and pharmacology, Univ. of Pittsburgh), prof. of physiology, in suc- cession to Dr. J. Van de Erve, resigned, now prof. of physiology, Marquette Univ. Univ. of Bonn : Dr. H. Seit er, acting director of the Hygienic Inst, succeeding Prof. Kruse. Univ. of Cal. : Dr. Frits W. Woll (Univ. of Wis.), prof. of animal 1913] General 115 nutrition- Scripps Inst, of Biolog. Research: Dr. F. B. Sumner, biologist. Univ. of 111. Med. Seh. (Chicago) : Dr. Geo. P. Dreyer, prof. of physiology; Mr. /. Craig Small, assis. in physiol. chemistry; Dr. Ber- nard Fantus, prof. of pharmacology ; Dr. Edgar D. Coolidge, prof. of materia medica and therapeutics. (See Bioch. Bull., 1913, ii, pp. 575 and 578.) Univ. of Leeds: Dr. Charles Crowther, prof. of agric. chemistry, in Charge of the work in animal nutrition; Dr. H. W. Dudley (Herter Laboratory), lecturer in biochemistry. Univ. of Leipzig: Dr. Walter Kruse (Bonn), prof. of hygiene and director of the Hygienic Inst., as successor to Prof. Hofmann. Univ. of Neb. Med. Seh. (Omaha) : Dr. Irving S. Cutter, prof. of biolog. chemistry; Dr. A. E. Günther, prof. of pharmacology. Univ. of Penn.: Dr. Howard B. Lewis (Sheff. Sei. Seh., Yale), instr. in physiol. chemistry (Seh. of Med.) ; Dr. Hermann Prinz, prof. of materia medica and therapeutics (Seh. of Dentistry). Univ. of Pittsburgh: Mr. Orville J. Walker, assis. in physiology and pharmacology. Univ. of Vienna: Prof. Julius Mauthner, director of the Medico- chemical Inst., vice Prof. E. Ludwig retired; Prof. Hugo Salomon, director of the med. clinic, vice Dr. Carl von Noorden resigned. Univ. of Wis. : Dr. Stephen M. Babcock, prof. emeritus of agric. chemistry; Dr. E. R. Miller (Ala. Polytech. Inst.) acting assis. prof. of plant chemistry. Western Reserve Univ. : Dr. Roy G. Pearce, instr. in physiology (promotion). Alvarenga prize. The Col. of Physicians, Phila., announces that the next award of the Alvarenga prize, being the income for one year of the bequest of the lata Senor Alvarenga and amounting to about $180, will be made on July 14, 19 14, provided that an essay w^orthy of the prize shall have been ofifered. Essays intended for com- petition may be on any subject in medicine, but cannot have been pub- lished. They must be typevv^ritten, and if written in a language other than English, should be accompanied by an EngHsh translation, and must be received by the secretary of the College on or before May i, 1 9 1 4. Each essay must be sent without signature, but must be plainly marked v^ith a motto and be accompanied by a sealed envelope hav- ing on its outside the motto of the paper and within the name and address of the author. It is a condition of the competition that the 1 16 Biochemical News, Notes and Comment [Oct. successful essay or a copy of it shall remain in possession of the College; other essays will be returned on application within three months after the award. Further information may be obtained on application to Thomas R. Neilson, M.D., sec'y, 19 S. 22d St., Phila- delphia. Grants. At the recent meeting of the British Assoc. for the Adv. of Science, at Birmingham, grants in aid of scientific research amounting to about $6,000 were made. The grants of special in- terest to biochemists are the following: Chemistry — Dr. W. H. Perkin, study of hydroaromatic substances, £15; Prof. H. E. Arm- strong, dynamic isomerism, £25 ; Prof. F. S. Kipping, transforma- tion of aromatic nitroamins, £15; A. D. Hall, plant enzymes, £25; Prof. W. J. Pope, correlation of the crystalline form with molecular structure, £25; Prof. H. E. Armstrong, solubility phenomena, £15. — Physiology: Prof. E. A. Schäfer, the ductless glands, £35; Prof. A. D. Waller, anesthetics, £20; Prof. /. S. Macdonald, calori- metric observations, £40; Prof. C. S. Sherrington, mammalian heart, £30. The Commit. of the Paris Acad. of Sciences, appointed to con- sider the distribution of the Bonaparte research fund, has made the following recommendations, among others, for 1913: R. Coquide, 2,000 francs, to assist him in his study of the turf lands of the north of France from the agricultural point of view; Paul Becquerel, 2,000 francs, for the continuation of his researches on the influence of radioactive substances on the nutrition, reproduction, and Varia- tion of some plant species ; M. Lormand, 2,000 francs, for the pur- chase of a sufficient quantity of radium bromid for methodical re- searches on the influence of radioactivity on the development of plants. Endowment funds. Foreign. Ernest Solvay, the discoverer of a process for the manufacture of soda, celebrated the fiftieth an- niversary of that discovery on Sept. 2 at Brüssels by giving more than $1,000,000 to educational and charitable institutions and the employees of his firm. The Universities of Paris and Nancy each received $100,000. — Dr. Gavin P. Tennent, of Glasgow, has be- queathed £25,000 to the Univ. of Glasgow, to be applied for such 1913] General 117 objects in connection with medicine as the trustees may determine. The Univ. has also received a legacy of £5,000 by the late Mr. William Weir, ironmaster, the income of which is to pay for an additional assist. to the prof, of materia medica. American. Mrs. Russell Sage has given $34,000 to Syracuse Univ., of which $30,000 is for the Joseph Slocum Agric. Col. — The General Educ. Board has announced a gift of $1,500,000 to the med. seh. of Johns Hopkins Univ., to be known as the William H. Welch Endowment for Education and Clinical Research, in recog- nition of Dr. Welch's distinguished Service to the cause of medical education in America. This is the greatest gift ever made by the board to a single Institution of learning. The proposed plan of spending the money opens the way for a new era in med. science. Briefly it is this : To so reorganize the med. seh. as to pay out of the income from the gift such salaries to the men who occupy the chairs of medicine, surgery and pediatrics (and to their assistants) as will enable them wholly to drop their private practices and devote their entire time, ability and lives to the advancement of their par- ticular branches. The departments which it affects are at present presided over by Dr. Lewellys F. Barker, prof. of medicine; Dr. W. S. Halsted, prof. of surgery; Dr. John Howland, prof. of pediatrics. Notes on radium. Appreciation of Madame Curie. All the World knows how Madame Curie ( Coming from Warsaw as Marie Sklodowska to work in Paris), inspired by the spontaneous radioactivity newly discovered by Becquerel, began in 1896 a met- rical examination of the radioactivity of minerals of all kinds ; and how, when a uranium residue showed a value larger than could have been expected from its uranium content, she, with exemplary skill and perseverance, worked down some tons of this material (given her by the Austrian government on the instigation of Prof. Suess), chemically dividing it and retaining always the more radio- active portion, until she obtained evidence first of a new dement which she christened polonium, in memory of her own country, and then after months of labor succeeded in isolating a few grains of the other and more permanent substance now so famous — a sub- stance which not only exhibits physical energy in a new form, but is ii8 Biochemical News, Notes and Comment [Oct. likely to be of Service to suffering humanity. Of the metallic base of this substance she determined the atomic weight, finding a place for it in Mendeleeff's series; and with the aid of her husband, whose lamentable death was so great a blow to science, she proceeded to discover many of its singular properties, some of them so extra- ordinary as to rivet the attention of the world. Subsequent workers engaged in the determination of numbers belonging to either of her special elements, radium and polonium, have sought her advice, and it has proved of the utmost value. (Sir Oliver Lodge, president of the British Assoc. for the Adv. of Science and principal of the Univ. of Birmingham, in introducing Madam Curie to the Univ. for the honorary degree of LL.D. : Science, 1913, xxxviii, p. 521.) National radium Institute. The Director of the Bureau of Mines authorizes the announcement that a cooperative agreement has been entered into with the newly organized National Radium Inst., whereby the Bureau obtains the opportunity of a scientific and technological study of the mining and concentrating of carnotite ores and of the most efficient methods of obtaining radium, Vana- dium and uranium therefrom, with a view to increased efficiency of production and the prevention of waste. The Institute was recently incorporated with the following officers : President, Howard A. Kelly, vice-president, Curtis F. Burnam, secretary and treasurer, Archibald Douglas, additional directors, James Douglas and E. J. Maloney. The Institute has no connection with the mining of pitchblende, details of which recently appeared in the Denver papers. It has, however, obtained the right to mine twenty-seven claims in the Paradox Valley region, among which are some of the best mines in this riebest radium-bearing region of the world. Nearly one hun- dred tons of high-grade carnotite have already been procured. Under the agreement with the Bureau of Mines, the technical Opera- tions of the mines and mill are to be guided by the scientific staff of the Bureau. Work will begin in an experimental plant to be erected in Colorado, using entirely new methods developed at the Denver Office of the Bureau of Mines. Concentration experiments also will be conducted in the Paradox, probably at the Long Park claims, and if successful will be applied to reducing the wastes that now 1913] General 119 take place. Within a year at most, the mill Operations should make results certain and the extraction of ore and production o£ radium will then be continued on a larger scale. The Separation of uranium and Vanadium will also be studied, a contract having already been signed for all of these by-products that may be obtained. All proc- esses, details of apparatus and plant, and general information gained will be published for the benefit of the people. The institute is supplied with sufficient funds to carry out its plans. It has been formed for the special purpose of procuring enough radium to conduct extensive experiments in radium therapy with special reference to the curing of Cancer. It also expects to carry on investigations regarding the physical characteristics and chemical effects of radium rays and hopes in time to be able to assist or perhaps even duplicate the effects of these rays by physical means. Actual experience, especially of the institute's president, in the application of the 650 mg. of radium and 100 mg. of mesothorium already in his possession, have led him and his associates to believe that with larger supplies many of the variables that can not now be controlled may be f ully correlated, and that radium may become the most effective agent for the treatment of Cancer and certain other malignant diseases. Important results have already been obtained by using high concentration of the gamma rays of radium with the alpha rays entirely cut off and the beta rays largely eliminated. Hospital facilities in both Baltimore and New York are already supplied. (Charles L. Parsons: Address to the i6th Annual Conv. of the Amer. Mining Congr., Phila., Oct. 20-24: Science, 1913, xxxviii, p. 612.) Important discoveries at the Radium Inst., London. From the Radium Inst, some important discoveries in radium therapy are announced. At the Inst, it has been demonstrated that radium emanation has exactly the same properties as pure radium and is as efficient for curative purposes. This is a discovery of the highest practical importance, for previously radium treatment could be given only at the Inst., as it was not practicable to lend this ex- tremely valuable and limited substance. Now the emanation fixed in a hollow plate or tube, is sent to physicians for use on patients. Thus, if a physician wants 200 mg. of radium for use on a patient, 1 20 Biochemical News, Notes and Comment [Oct. its cost, $20,000, woiild be prohibitive. But. for a comparatively trifling sum the Inst, can supply a plate containing radium emana- tion which will have the same effect. There is this difference, how- ever, the activity of the emanation decreases, falling to one-half strength in three and one-half days. At present, i gm. of radium is devoted wholly to producing emanation for distribution, and as the demand is so great, 1.5 gm. is about to be used. Another branch of the activity of the Inst, is the supply of water impreg- nated with radium emanation for consumption in certain affections. The Inst, is supplying radium-emanation Solution of a strength of from I to 2 millicuries per liter. The Radium Inst, was opened in Aug., 191 1, and since that time the work has steadily increased. At first it was open from 8 a. m. to 6 p. m. for the purpose of treating patients. So numerous were the poor patients that a night clinic had to be added sixteen months ago. It is open until 11.30 p. m., and sometimes until midnight. . . . During the month of August the Inst, was closed in order that mem- bers of the staff, who were working at high pressure and all of whom have radium burns on their hands, might have a holiday and rest, which is the only known eure for these burns. The quantity of radium in the Inst, is, at the present price, of the value of $400,000, and amounts to 4 gm, (London letter: Jour. Amer. Med. Assoc, 1913, Ixi, p. 1469.) Dangers of radium. The assist. med. sup't, Dr. Arthur Bur- rows, of the Radium Inst., London, states that most of the staff have been burned to a greater or less extent at some time or another. In his own case he found the skin peeling off his fingers when he went to play golf. The nurses, however, who do most of the actual handling, suffer most. In addition to the more or less pain- less skin-peeling, the finger nails become brittle and split down the Center, ulcerated Spots appear, and in time the hands become totally anesthetic. It is curious that the hands of those who have much to do with radium are always far more susceptible to heat than to cold. Gloves are not much protection. The only thing to do when the fingers show these Symptoms is to have nothing to do with radium until they recover. Those who develop burns are usually given some work in connection with the Inst, which does not involve 1913] General 121 immediate contact with the dement. Radium in course of time burns most things with which it comes in contact. For instance, the Hning of the boxes in which it is kept is often entirely eaten away. The ill effects are not feit in the human body until a fort- night after the contact. It eats away the abnormal tissues, such as Carcinoma, sarcoma, etc., and leaves the surrounding normal tissues in an ordinary condition. In its antipathy to abnormal tissues lies its curative properties in these cases. But in time, or as the result of excessive application, radium will have an effect also on the normal tissues. A subsidiary effect on the patient is increased sus- ceptibly to changes of temperature over areas that have been treated with radium. Many patients who have had rodent ulcers and super- ficial skin lesions, cured with radium, experience great discomfort at the site of the old lesion when very cold or very warm air plays on it. This susceptibility, however, gradually disappears in two or three months. A marked condition of lethargy is frequently, it might almost be said invariably, noted in patients receiving pro- longed exposures with large quantities of heavily screened radium. It generally makes its appearance about the fourth day of the treat- ment, and passes off within a few days of the cessation of the ex- posures. (London letter: Jour. Amer. Med. Assoc, 1913, Ixi, P- I549-) MuNiciPAL owNERSHip OF RADIUM. On favorablc reports as to the therapeutic effects of mesothorium in can'^er, the communal authorities of Essen have determined to purchase 200 mg. of the preparation. Half of the necessary sum, $10,000, has been raised by private subscription and the rest has been appropriated by the communal authorities. — A bureau for the distribution of radium and mesothorium has been f ounded in the Hamburg Inst, for Cancer and Tuberculosis Research, which was founded a short time ago. The object is to secure as large a quantity of these preparations as possible in a short time and place them at the disposal of the public. At present about 150 mg. of radium bromid are on band; this quantity is to be doubled in about two weeks and there is a prospect of securing further amounts. The preparations are to be loaned to physicians. — The favorable results which have been obtained with mesothorium radiations in Carcinoma by the gynecologists Bumm, 122 Biochemical News, Notes and Comment [Oct. Krönig and others have excited great attention in our newspapers and partly under the pressure of public opinion and partly instigated by the wishes of the directors of the hospitals, the municipal authori- ties in a number of cities have determined to purchase some meso- thorium and radium. BerHn has appropriated $50,000 for this purpose, and $200,000 have been appropriated for the same purpose by the Prussian Department of Education. It is to be hoped that further success will justify this not inconsiderable material sacrifice. — The great rush for the purchase of mesothorium and radium by municipalities, has been suddenly checked by the city of Munich. The city government of that city has refused for the present to carry out the resolution to buy $50,000 worth of this costly material. It is believed that there are positive evidences that the factories engaged in producing mesothorium are raising the price unduly. For this reason, more exact information is to be obtained by the municipal authorities before the purchase of the preparation is con- summated in Munich and other cities. (Berlin letter: Jour. Amer. Med. Assoc, 1913, Ixi, pp. 613, 1055, 1308 and 1470.) The Prussian Government has purchased a gram of radium at the cost of $87,500 for hospital and scientific use. The Prussian ministry of education, which a short time ago made grants of money to the univ. clinics at Berlin, Halle and Kiel, enabling them to procure radium or mesothorium for the treatment of Cancer, is now said to have placed $200,000 in the estimates of next year for further purchases. There v^as a section of radiology at the last Intern. Congr. of Med., for the first time in the history of the congress. Meetings of societies and congresses. British Assoc. for THE Adv. of Science, The annual meeting of the British Assoc. for the Adv. of Science was held in Birmingham. The attendande numbered 2,500. Hormones. In the Sect. of Physiology the most important paper was that on internal secretions, by Prof. Schäfer. He pointed out that the convenient term hormone, introduced by Starling (from op/xaco, I excite) while applicable to the active principle of many internal secretions, has been extended to all and is wrongly I9I3] General 123 applied to principles which do not excite, but check activity. For these he suggested the term chalone ( f rom x<^^^^' I relax) . It is, however, desirable to have a term which includes both the hormones and chalones. The one quality which distinguishes them is their drug-like effect on the organs and tissues. A convenient term, suggested by Prof. W. R. Wardie, is autacoid substance. Discussion of the origin of life. A large audience attended a combined meeting of the sections of physiology, zoology and botany for a discussion on the origin of life. At this meeting the subject was introduced by Dr. B. Moore, prof. of biochemistry in the Univ. of Liverpool. He regarded the problem as an experimental one and Said that he could demonstrate a step which connected inorganic with organic matter. The world of living plants and animals de- pends on the synthesis of organic from inorganic Compounds by the chlorophyl of plants acting as a transformer of light energy into chemical energy. This State of affairs must have evolved from something more simple, for chlorophyl is one of the most complex of known organic substances. In considering the origin of life the Start must be made in a purely inorganic world. As the results of eighteen months' experimental work, he believed that he has ob- tained evidence of the first step in organic evolution. When dilute Solutions of colloidal ferric hydroxid or the corresponding uranium Compound are exposed to strong sunlight, there are ."vnthesized the same Compounds as are formed in the first stage of organic syn- thesis by the green plant — formaldehyd and formic acid. If now they considered a planet cooling down and exposed to sunlight, at first Clements only would be present. As it cooled, binary Com- pounds would form and then simple crystalloid salts. By the union of Single molecules into groups of fifty or sixty, colloidal aggregates appeared. As these increased in complexibility they became more delicately balanced (labile). They were easily destroyed by sudden changes in environment, but within certain limits were peculiarly sensitive to energy changes and could take up energy in one form and transform it into another. These labile colloids took up water and carbon dioxid and, utilizing the sunlight Streaming onto the plant, produced the simplest organic structures. Next these struc- tures reacting with themselves and with nitrogenous inorganic mat- 124 Biochcmical News, Notes and Comment [Oct. ter, continued the process and built up more and more complex and also more labile organic colloids, until finally these acquired the property of transforming light energy into chemical energy. In the discussion which followed, Sir Oliver Lodge agreed that new possibilities entered matter with the increase of size and com- plexibility of the molecule. A molecule sufficiently complex and sufficiently unstable and supplied with energy by the sunlight ap- parently gave the chemical substratum for the Operations of life. It was Potential living matter. This has not been made yet, but he has not miich doubt that it might be done. To produce potential living matter, however, is not to produce life. He regards life as of a higher order, for he does not consider the universe limited entirely to what we know. Professor Armstrong said that as a chemist he is not for a moment prepared to accept Schaf er's contention that it is probable that we shall ever be able to produce life. This would mean a series of Operations so infinitely complex that it is not within our power to pronounce any opinion on its possibility. The dominant Word in Moore's paper was the word " colloid." It is a blessed word among the physiologists at the present day, but like so many blessed words is used for wrapping up ignorance. Professor Hartog said that there was a tremendous amount of scientific "bluff" in the assertion that there was a consensus of opinion among biologists that life was only one form of chemical and physical action which could be produced in the laboratory. The greatest biologists held aloof from such dogmatism. (London LETTER: Jour. Amer. Med. Assoc, 1913, Ixi, p. 1307.) Intern. Med. Congr. The Intern. Med. Congr. (17) was formally opened at Albert Hall, London, Aug. 6, by Prince Arthur of Connaught. Ehrlich on chemotherapy. At the general Session on Friday, Aug. 8, Professor Ehrlich delivered, in German, the address in path- ology taking for his theme : Chemotherapy. After referring to the work of Jenner, Lister, Sir Patrick Manson, Ross, Castellani, Bruce, Leishman and others on the protozoan diseases, he entered into a technical explanation of the principle of chemotherapy, especially with reference to the work done in elaborating salvarsan. He ex- 1913] General 125 plained that salvarsan has not only a direct parasiticidal action, but that immunity of parasites to such action could be accounted for only by a purely chemical diminution of their affinity; and a com- plete exhaustive knowledge of the various chemical peculiarities of a parasite, which he called the "therapeutic physiology of the para- sitic cell," is essential for its successful chemotherapeutic treatment. Certain chemical peculiarities are found in many different kinds of parasites. In proportion as more of these chemical afünities are discovered, the greater is the possibility of successful chemotherapy. He still keeps in view the idea of freeing the body of micro- organisms by one or at most two injections of the proposed remedy, and in his animal experiments this principle is still being pursued. He looks forward to the extension of the principle of chemotherapy as a means of bridging the gaps which still exist in our knowledge of the treatment of some diseases. In the diseases involving pro- tozoa and spirilla, good results have already been gained. In a series of other diseases, such as small-pox, scarlatina, typhus, and perhaps also yellow fever, but above all the infectious diseases caused by invisible germs, there is a bright prospect of success. In the common bacterial diseases due to Streptococcus, staphylococcus, and the micro-organisms of typhoid, dysentery and tuberculosis, he feels that the struggle is a hard one, but that success in these dis- eases will also be attained on the principle of chemothtrapy. (Lon- don LETTER: Jour. Amer. Med. Assoc, 1913, Ixi, p. 610.) On the " art of conciseness." The program was generally over- crowded and Speakers often raced the clock to get in what they wanted to say in the allotted time, and were brought to a premature end by the chairman's bell. As in all medical gatherings, the in- capacity of even those who were eminent and had something to say, to say it properly and concisely, was painfully evident. The fifteen minutes allotted to a Speaker, if properly used, was in most cases amply sufficient for the presentation of his conclusions and his reasons for them, but want of conciseness of expression as well as want of judgment in suppressing unnecessary details prevented this. Instead of brief but sufficient general description, worse than useless details which only wearied the audience were presented. It is curious that no one seems to trouble about the reform of this uni- 126 Biochemical News, Notes and Comment [Oct. Versal evil. The man who could compel the education of medical authors in the art of conciseness, before they be permitted to speak or write, would deserve a place in history among the benefactors of humanity. (London letter: Jour. Amer. Med. Assoc, 1913, xli, p. 612.) Amer. Chem. Soc'y. The annual meeting (48) of the Amer. Chem. Soc'y was held at Rochester, N. Y., Sept. 8 to 12. This was the first meeting in Sept. under the newly adopted Constitution. The large number present and the enthusiasm of the meeting amply justify the change in date from the Christmas holidays to the fall of the year, Dr. Charles L. Parsons was re-elected secretary of the society, and Dr. A. P. Hallock, treasurer, for a period of three years, under the revised Constitution. Prof. W. A. Noyes was re-elected editor of the Jour. of the Amer. Chem. Soc'y, and the board of associate editors was continued, with the exception of Drs. H. P. Talbot and A. A. Noyes, who asked to be relieved of this duty. Prof. W. Lash Miller, of the Univ. of Toronto, was elected to the board with special reference to physical chemistry. Prof. M. C. Whitaker was re-elected editor of the Jour. of Indus, and Eng. Chem., and the board of associate editors was continued and the editorial staff strengthened by the addition of two assist. editors. Prof. A. M. Patterson was re-elected editor of Chemical Abstracts, and Drs. /. /. Miller and E. J. Crane assoc. editors. ( See page 76. ) Intern. Congr. of Refrigeration. The Third Int. Cong. of Refrig., with an attendance of nearly 2,000 delegates, over 400 of whom were from abroad, convened in Washington and Chicago, from Sept. 15 to Oct. i. The officers of the Third Section (on the " application of refrigeration to f oods for the purpose of conserving and preserving them") were: President, Dr. Harvey W. Wiley; vice-pres., Mr. C. H. Parsons; sec'y, Dr. M. E. Pennington; addi- tional member of the sect. commit., Prof. R. M. Allen, Prof. H. J. Eustace, Mr. H. C. Gardner, Prof. Wm. J. des, Mr. /. L. Hughes, Prof. W. A. Stocking, Prof. A. V. Stubenrauch, Mr. R. H. Switzler. Miscellaneous items. Lane lectures. The f ourteenth course of Lane med. lectures was delivered in Lane Hall, San Francisco, 1913] General 127 on the evenings of Sept. 3, 4, 5, 8 and 9, by Prof. Sir E. A. Schäfer, pro f. of physiology, Univ. of Edinburgh, on Internal secretion in general, The thyroparathyroid glands, The adrenal glandulär appa- ratus, The pituitary body, The influence of internal on other secre- tions. Prof. Schäfer also delivered at Stanford University a lec- ture on Methods of resuscitation. Pasteur Inst, twenty-five years old. Owing to the renown of Pasteur's studies of rabies, an international subscription, which was opened by the Acad. des Sciences de Paris, soon amounted to $500,000 and permitted the foundation, twenty-five years ago (Nov. 18, 1888), of the Institut Pasteur. At present the Inst, is a center at once of scientific research, of higher instruction and of thera- peutic treatment. It is divided into three principal sections : micro- biologic, serotherapeutic, and biochemical. One of the sources of superiority of the Pasteur Inst, consists in its independence. It was founded and is carried on without ofiicial superintendence and hence has a spirit of initiative and of adaptiveness which administrative oversight scarcely permits to government establishments. ScHOOL FOR Public Health Officers. Harvard Univ. and the Mass. Inst, of Tech, will cooperate in maintaining a Seh. for Public Health Officers. Prof. M. J. Rosenau (Harvard) is the director. The work of the school will be under his immediate supervision, in association with Profs. W. T. Sedgwick and Geo. C. Whipple as an administrative board. ApPRECIATION of the SERVICES OF THE COMMIS. ON ElECT. Shock. At the last annual meeting of the National Elect. Light Assoc, the following resolution was unanimously adopted: Whereas, The Assoc. has accomplished a most creditable piece of humanitarian work in the issuance of its rules on resuscitation from electric shock used throughout the world and approved formally by other industries, the national government and State boards; therefore, be it Resolved, That the thanks of this association be extended to the Med. Commis. for its splendid results, and also to the Amer. Med. Assoc, without whose active Cooperation these laudable results could never have been achieved. The work of this commis. is monumental and its effects will be 128 Biochemical News, Notes and Comment [Oct. widespread. It is an excellent illiistration of the valuable results which can be secured through practical cooikration between the medical profession and enlightened business men for the saving of Hfe and the prevention of accidents. (Editorial: Jour. Amer. Med. Assoc, 191 3, Ixi, p. 1637.) The commis. consists of Prof. Walter B. Cannon, chairman; nominated by the Amer. Med. Assoc, Prof. Yandell Henderson, Dr. George W. Crile, Dr. 6^. /. Meltzer, nominated by the Nat. Elec. Light Assoc, Prof. Edward A. Spitnka, Mr. W. C. L. Elgin, nomi- nated by the Amer. Inst, of Elec. Engineers, Prof. Elihii Thompson, Dr. Arthur E. Kennelly, Mr. W. D. Weaver, sec'y (elected by the commis.). Ether day. The sixty-seventh anniversary of Ether Day was celebrated in the lower amphitheater of the outpatient's dep't of the Mass. Gen. Hosp., Oct. 16. The principal address was dehvered by Dr. M. J. Rosenau. Carbates. In this age of method, accuracy and conciseness, we may say sulphates instead of sulphurates ; phosphates for phos- phorates (better still, sulfates and fosfates) ; nitrates for nitro- genates ; chlorates for chlorinates. Why should we not say carbates instead of carbonates? We already say carbides instead of car- bonides ; why should we not f ollow the f ashion consistently and say carbates f We should then have the word carbation to mean the formation of carbates, leaving the word carbonation to refer to the development of carbon in a substance which would fittingly corre- spond to the present word carbonize, and so avoid a puzzling am- biguity. Furthermore, the saving of time and printer's ink would amount to something in a word so often used. (J. E. Todd: Science, 1913, xxxviii, p. 270.) Mark crucibles with ink. It is a more or less common practice to mark porcelain crucibles or other articles with ordinary fountain pen ink. The usual directions are to dry and subsequently heat in the blast, repeating the whole Operation as often as neces- sary to obtain a clear brown figure. It is a saving of considerable time if a convenient area of the crucible first be heated in a Bunsen flame and the figure then drawn with a fountain pen. The ink 1913] Columbia University Biochemical Association 1 29 dries iiistantaneously, and in one Operation leaves a coating of any thickness desired. Such figures are distinct even when exceedingly small, and have the advantage of being practically permanent. (Ross Allen Baker: Chemist- Analyst, 1913, Aug., p. 12.) II. COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION I. General notes Marriages : On July 20, Miss Jessie Archibald Moore and Dr. Alfred Henri Rahe. — On Sept. 25, Dr. Martha Ornstein and Dr. J. Bronfen Brenner. Engagement: Miss Eleanor Riehm, of Newark, N. J., and Dr. Clayton S. Smith. Appointments.^ Dr. Chas. F. Bolduan, lect. on hygiene and sanitation, N. Y. Univ. and Bellevue Hosp. Med. Col. — Dr. Sidney Born has been retained by Prof. M. C. Whitaker to conduct an ex- tensive investigation of the composition and constituents of certain vegetable oils, with particular bearing on the organic substances oc- curring in extremely small proportions. — Dr. E. D. Clark (instr. in chemistry, Cornell Univ. Med. Seh.), soil biochemist, Bur. of Chemistry, U. S. Dep't of Agric. — Dr. C. B. Coulter, assis. in pathology, Columbia Univ. — Mr. Fred D. Fromme, assis. in botany, Indiana Exp. Station. — Dr. T. Stuart Hart, assis. prof. of clin. medicine, Columbia Univ. (promotion). — Dr. Homer D. Hoiise (assoc. director and lect. on botany and dendrology, Biltmore Forest Seh.), assis. State botanist of New York. — Dr. Emile F. Krapf, research chemist and chief of the pharmaceut. dep't, Radium Re- search Lab. of the Standard Chem. Co., Pittsburgh. — Dr. Max Morse (prof. of biology, Trinity Col.), instr. in biochemistry, Univ. of Wis. — Dr. Reuben Ottenberg, instr. in bacteriology, Columbia Univ. — Mr. P. W. Punnett, assis. in chemistry, Columbia Univ. — Dr. Jacob Rosenbloom (assis. prof. of biochemistry, Univ. of Pitts- burgh), biolog. chemist, Western Penn. Hospital, Pittsburgh. — Dr. Charles Hendee Smith, instr. in diseases of children, Columbia Univ. 1 See footnote, page 113. 130 Biochemical News, Notes and Comment [Oct. Investigators at Woods Hole. The following members of the Assoc. were among the investigators, during the past summer, at the Marine Biolog. Lab., Woods Hole, Mass. : Cora J. Beckwith, A. J. Goldfarb, H. B. Goodrich, Mildred A. Hoge, Louise H. Gregory, E. N. Harvey, R. R. Hyde, Jacques Loeb, Max W. Morse, Charles Packard, A. M. Pappenheimer, A. Franklin Shull, C. R. Stockard, Hardolph Wasteneys, Isabel Wheeler, L. L. Woodruff. Officers in societies. Dr. Carl L. Aisher g has been appointed secretary of the Assoc. of Official Agric. Chemists. — Dr. Raymond C. Osburn has been elected president of the N. Y. Entomolog. Soc'y. — Dr. Ralph G. Stillman is president of the Nu Sigma Nu Alumni Assoc. and Drs. Clinton B. Knapp and Wm. K. Terriherry are members of the exec. commit. — Dr. Philip Van Ingen is a member of the Public Health Hosp. and Budget Commit. of the N. Y. Acad. of Med. Lectures. Dr. Jacob Rosenbloom recently delivered a lecture before the Scientific Soc'y of the Western Penn. Hosp., Pittsburgh, on the evolution of modern biochemistry. At this Institution he is also giving a course of lectures, with demonstrations, on methods for the analysis of urine, feces, stomach contents and breast milk (five lectures), nutrition (ten lectures), and the chemistry of plant and animal life (five lectures). — At the tenth anniversary of the establishment of the Desert Lab. of the Carnegie Institution, which was celebrated at Tucson, Ariz., Sep. 20, Prof. B. E. Livingston demonstrated to the visitors, including members of the Intern. Phytogeographic Soc'y, some of the results of his researches, now in progress, on Water relations of plants. — Dr. F. J. Seaver de- livered one of the "late summer lectures" at the N. Y. Botan. Garden, on Shade trees and their enemies. Doctorates. Miss Marguerite T. Lee and Mr. Sidney Born recently passed public examinations in final completion of the re- quirements for the Ph.D. degree at Columbia Univ. The subjects of their dissertations were, respectively, A study of modifications of the biuret test, and The chemical Constitution of invertase. Miscellaneous items. Prof. R. Burton-Opitß has been elected President of Alpha Omega Alpha, the honorary medical society, which now has chapters in the seventeen leading medical schools. 1913] Columbia University Biochemical Association 131 Miss Helen Gavin recently completed the requirements for the M.A. degree in biolog. chemistry at Columbia Univ. Dr. A. J. Goldfarh continued, during the summer at the U. S. Fisheries Marine Biolog. Sta. (Beaufort, N. C), his experiments on the grafting of eggs, and on changes in organisms produced by chemical means. Prof. Wm. H. Woglom is the author of Vol. I of Studies in Cancer and Allied Subjects, conducted under the auspices of the George Crocker Special Research Fund at Columbia Univ. The volume is entitled : The study of experimental Cancer : A review (1913, pp. 288). Dr. Rhoda Erdmann, of the dep't of protozoology of the Berlin Inst, for Infec. Dis., has been appointed Seessel research fellow in zoology at Yale Univ., to enable her to study Prof. L. L. Woodruff' s pedigreed race of Paramaecium. 2. Proceedings of the Association The third annual dinner will be given, at the Hotel Marseilles, on November 21. The guest of honor will be Prof. Lafayette B. Mendel, who will address the Assoc. on the results of his studies of Growth. The f ourteenth scientific meeting will be held at the Col. of Phys. and Surg., on Dec. 5, at 4 p. m. The proceedings of each of these meetings will be published in the January issue of the Bulletin. 3. Columbia Biochemical Department Marriage: On Sep. i, Miss Ethel Brand and Dr. Louis E. Wise. Resignations from and appointments to the staff . Resigna- TiONS. Dr. Max Kahn (assoc.) has been appointed an assist. in the pharmacol. lab. of the U. S. Bur. of Chem. — Dr. Louis E. Wise (instr.), has been appointed instr. in chemistry at the Univ. of Missouri. Appointments. Dr. Sergius MorguliSj lately research fellow in the Nutrition Lab. of the Carnegie Inst. (Boston), has been ap- pointed instr., to succeed Dr. Louis E. Wise. — Mr. Arthur Knud- 132 Biochemical News, Notes and Comment [Oct. son, who served temporarily as chemist during the summer in the Turck Inst. (N. Y.), has been reappointed to his assistantship in this laboratory, Miscellaneous items. Prof. Gies, on Oct. 11, delivered the second of the nine " autumn lectures " at the N. Y. Botan. Garden in a series on foods, subject : The digestion of vegetable foods. Prof. Gies was associated with Drs. L. H. Baekeland and R. E. Doohttle as an appointed representative of the Amer. Chem. Soc'y in the N. Y. Gen. Commit. for the entertainment of the foreign delegates arriving at the port of New York to attend the Third Int. Congr. on Refrigeration (p. 126). Volume III of " Studies in Cancer and alhed subjects, under the auspices of the George Crocker Special Research Fund at Columbia Univ.," was recently issued. It contains reprints of papers from the departments of zoology, surgery, clinical pathology and biolog. chemistry. "Part IV: Dep't of biolog. chemistry," which was edited by Prof. Gies, occupies pages 151-295 and consists of papers by Prof. Gies and Drs. Max Einhorn, S. S. Friedman, Max Kahn, Morris H. Kahn, D. J. Kaliski, Reuben Ottenberg, Jacob Rosen- bloom, Charles H. Sanford, and J. W. Weinstein. EDITORIALS The average scientist has often wondered how logic, with that diametrically opposed to it, can together find such a comfortable resting-place in the mental abode of Sir O. Lodge. This renowned Sir Oliver Lodge on English man of science, in his recent presidential "Continuity" address before the British Assoc. for the Adv. of Science/ advances at one and the same time ideas very plausible and others highly improbable, to say the least. That the laws of chemistry and physics hold sway in the animate as well as the inanimate world, but that the animate is something more than a mere conglomeration of chemical and physical laws, seems highly consonant not only with reason but with Observation. But why Sir Oliver should put faith in psychic phenomena — the study of which thus far has been barren of any tangible result — as a means of supplying the missing link in "continuity," is beyond comprehension. " Ever since the time of J. R. Mayer," writes Sir Oliver, " it has been becoming more and more certain that, as regards Per- formance of work, a living thing obeys the laws of physics, like everything eise; but undoubtedly it initiates processes and pro- duces results that without it could not have occurred — from a bird's nest to a honeycomb, from a deal box to a warship. The be- havior of a ship firing shot and shell is explicable in terms of energy, but the discrimination which it exercises between friend and foe is not so explicable. . . . Life introduces something incalculable and purposeful amid the laws of physics; it thus distinctly Supplements those laws, though it leaves them otherwise precisely as they were, and obeys them all." Thus far, thus good! Loeb or Schäfer might be tempted to deny part of this Statement, or Supplement it, but for most of us it seems to have the ring of truth. But what are we to make of this : "... the f acts examined have convinced me that memory and 1 For a complete account see the London Times, Sept. ii, 1913. 133 134 Mathews Plan for American Biological Society [Oct. affection are not limited to that association with matter by which alone they can manifest themselves here and now, and that per- sonality persists beyond bodily death." What " facts " is Sir Oliver speaking of ? To the scientific World at large these " facts " have thus far proved delusions pure and simple. Not only has not a single " f act " been substantiated, but overwhelming evidence is accumulating daily to show how utterly preposterous are assertions of this kind as to known facts. True, Sir Oliver, there may be more in this world than is dreamt of in our philosophies, but our present methods of studying psy- chical phenomena give no promise whatsoever of bringing those dreams v^ithin ränge. Our limited vision and our limited capacities are the only weapons with which we can fight life's battles. But limited as we are, need there be any cause for pessimism? Who knows what further evolution in man, as well as further develop- ment of the sciences, will bring. B. Horowitz. In pursuance of our plan to facilitate open consideration and possible removal of the obstacles in the way of more effective bio- logical Organization in this country, we append a few additional The Mathews plan quotations from letters on the subject^ and also for an American present, in the succeeding editorial by Dr. Eddy, Biological Society g^ summary of the opinions published in this num- ber, and in the April and July issues of the Bulletin. Wm. N. Berg, Bureau of Animal Industry, Washington, D. C. Most of my thoughts on Dr. Mathews' plan of organizing an Amer. Biolog. Soc'y and of lowering the cost of a collection of Journals of biological science have already been expressed by those of your cor- respondents who oppose the formation of any new Organization on the ground that there are societies enough at present. Revision down ward, if anything, is wanted. Any further federation of existing biological societies that would result in the benefits of Co- operation would naturally appeal to all of us that are interested. ^We received a number of letters in which the authors indicated an Inten- tion to give the plan mature deliberation before expressing their opinions. In most of these cases the expected comment has not been received. 1913] Editoriais 135 I am in favor, however, of that part of the plan pertaining to the lowering of the subscription rate on several Journals if all are sub- scribed for at the same time. This would enable an instructor located, let us say, in the middle West or South, whose nearest libraries are of moderate size, to supply himself with the Journals he needs. But in so far as men nearer large libraries might find this unnecessary, I believe a vote should be taken so that those who are willing to subscribe to a set of Journals can express their wishes in the matter. Speaking for myself — " I couldn't drink another drop." Ulric Dahlgren, Princeton, N. J. I would prefer to see the existing Soc'y of Naturalists continued and strengthened and re- formed. Lewis W. Fetzer, Office of Expt. Stations, U. S. Dep't of Agric, Washington, D. C. I am of the opinion that the Mathews plan is an admirable one, but the success of the Organization can only come through the amalgamation of existing societies. An entirely new Organization is an unnecessary evil. The abstract Journal idea is a good one and should receive the generous support of all interested in the biological sciences. To say the least, we are sorely in need of a Journal of this character. Members of the new society should have the option of subscrib- ing to three or more Journals, but the abstract Journal should always be amongst the minimum number. This would allow investigators with limited finances to subscribe to a few foreign Journals. A "Journal of Biological Industries" is needed in this country, but it should not be forced upon all the members of the society. William J. Gies, Columbia Univ. I favor the objects of Prof. Mathews' plan. I believe that the logical development of the Feder- ation of Amer. Societies for Exp. Biology, which was organized a year ago and which is in effect an embryonic Amer. Biolog. Soc'y, would secure all the many desirable results at which Prof. Mathews' excellent and far-reaching plan is aimed. The Federation, by a process of evolution, will probably gather into its affiliations all the societies that show natural tendencies to Cooperation ; the constituent societies will be natural sections; the obvious economies in the issuance of Joint programs and the publication of coordinated pro- 136 Mathews Plan for American Biological Society [Oct. ceedings will suggest others ; and tlie advantages of labile Organiza- tion of independent societies in natural interdependent relationships will impel careful consideration of such additional projects as the evolution of the biological sciences may suggest. I think we should proceed as rapidly as possible in the direction of the Mathews plan through the agency of the Federation — that we should perfect the latter Organization and go forward with such further developments as the growth of the Federation might suggest and determine.^ Philip B. Hawk, Jefferson Medical College, Phila. The Mathews plan for the Organization of an Amer. Biolog. Soc'y ap- pears to me to have much to commend it. Joseph S. Hepburn, Food-Research Lab., U. S.Dep'tof Agric, Phila. While a federation of the various biological societies may be consummated, their complete merger is a rather remote possi- bility. Thus the chemical engineers, the electrochemists, and the biological chemists have their separate organizations, entirely inde- pendent of the Amer. Chem. Soc'y; the biological societies aremore numerous than the chemical, and their complete merger would be a more difficult proposition. A membership fee of $20 or $30 for the new society would perhaps be prohibitive to many biochemists who are already members of the Amer. Chem. Soc'y, and also pay membership fees in one or more local scientific societies, Institutes or academies, in order to gain access to the complete files of scien- tific Journals, foreign and domestic, in the libraries of the latter in- stitutions. Moreover, the average biochemist would constantly use perhaps three, and occasionally perhaps as many more, Journals in the list of fourteen; the others would be of very little use to him. In this connection it should be remembered that the Amer. Chem. Soc'y does not supply to its members all of the chemical Journals published in the United States. The proposition to grant member- ship and subscriptions to say any three or four Journals out of the Hst of fourteen for a fee of about ten dollars would doubtless make a strong appeal to the biochemist. The biological abstract Journal is a gigantic undertaking, since it should include, in addition to abstracts of papers in the various branches of botany and zoology, the material now found in the 1 Editorial : Biochem. Bull., 1913, ii, p. 332. 1913] Editoriais 137 Zentr. f. Biochem. u. Biophys., Jahr, über die Fortsch. d. Tierchem., and Centr. f. Bakter., Parasitenk. u. Infectionskr. The financial bürden of Publishing such a Journal would be so great that it would have to be limited in scope, for instance leaving biochemical ab- stracts to Chem. Ahstr., and thus depriving members of the new Society, not subscribers for the latter Journal, of abstracts of a large portion of biological science. The financial dilemma might also be solved by Converting the new Journal into an index of biological literature, which would give the titles but not the subject matter of researches in all the fields of biology. Local sections of the new society or federation, sufficiently broad in scope, would fill a long- feit need, even in the very large cities ; existing local societies might well serve as nuclei which, by a proc- ess of expansion, could develop into local sections. Paul E. Howe, Columbia Univ. The Mathews plan for the amalgamation of the various existing societies for the promotion of the biological sciences is most attractive as an ideality. The practical attainment of the plan, as suggested, appears feasible. The estimates might be questioned when we consider the budget of the Amer. Chem. Soc'y, with its large membership and the present indication that the membership fee may be increased. In addition to the advantages of affiliation, the matter of an ab- stract Journal is most attractive. The cost of maintaining such a publication which would be entirely satisfactory, without presum- ing the use of another to Supplement it, would, it seems to me, be much greater than is estimated. One possibility has suggested itself for the reduction of expense and the economical attainment of a satisfactory biological abstract Journal : Cooperation with the Amer. Chem. Soc'y, so that the members of the biological society would receive Chem. Abstracts with its biological section, which would permit the biological society to confine itself to the publication of abstracts in fields not covered by Chem. Abstracts. In general I am most heartily in favor of the proposed society with its accom- panying advantages. Max Kahn, Bureau of Chemistry, Washington, D. C. I am fully in accord with Dr. Mathews' plan for the Organization of an Amer. Biolog. Soc'y. A Biolog. Abstr. Jour. is a necessity at the 138 Mathews Plan for American Biological Society [Oct. present time. There is no Journal that adequately and completely reviews all the current biological literature, and the biologist must himself plod through all the biological Journals in the languages which he can read, and trust to luck that in the periodicals which he has not examined there is naught of interest for him in the special biological field that he may be working in. I have had occasion to look through most of the abstract Journals in medicine and in chem- istry, and I have found them all wanting. The Jour. of the Royal Micros. Soc'y omits all papers of biochemical nature, and usually treats only of those biological papers which deal with morphology. Alfred P. Lothrop, Columbia Univ. The plan suggested by Prof. Mathews is an admirable one provided the autonomy and Organization of the existing societies are preserved in the new Or- ganization. In other words the mere payment of dues should in- clude membership in the general society, but the sections (the exist- ing societies) should be entirely free to elect into their membership such members of the general society as can present the qualifications required for membership in the existing societies. A biological abstract Journal would be of immense value and the Biolog. Sect. of Chem. Ahstr. might well be turned over to the management of the proposed " Biological Abstracts." A plan of a scale of fees to include the abstract Journal and as many other publications as might be selected would seem to be more feasible than levying dues large enough to include subscriptions to all the Journals on the list. S. S. Maxwell, Univ. of California. I have taken time to give considerable thought to the Mathews plan before expressing an opinion. It now seems clear to me that, notwithstanding the good features of the proposal, the result would be an additional Journal and an additional society, and that thus the bürden would not be lifted but made heavier. Amos W. Peters, The Training School at Vineland, N. J. Any serious consideration of the Mathews plan at once raises several im- portant questions. Is the proposed Organization desirable in addi- tion to those now existing, or, if it is to absorb them by what is essentially an extension of the present Federation plan, would it be desirable to extend the process of federation so as to include all biological organizations ? In other words, are the unity of inves- 1913] Editoriais 139 tigative method and the viewpoint of data and the breadth of inter- est of those who represent widely different subjects, which however all deal with a common living matter — are these sufficiently devel- oped today to hold such an Organization together? The native, inherent correlations of these subjects and their irresistible move- ment towards ultimate quantitative physical and chemical method must come home at times to every investigator in these fields. If the biological subjects shall be held together by one comprehensive Organization the bond of union w^ill have to be more effective than that which is represented by our greatest example of combination, the A.A.A.S. Why has the Amer. Chem. Soc'y, our model for the proposed biological society, grown up aside from and after the complete Organization of the A.A.A.S.? Are the divisions of biology as now pursued naturally articulated so that the analogy with chemistry holds or would they be conglomerate units so as to be analogous to the A.A.A.S. ? The probably correct opinion is that biology as a whole today occupies a middle position in this respect. In the last analysis it is probably largely a question of whether leadership, Cooperation and the practical conduct of such an Organ- ization can stimulate and maintain the interest of its constituents. The publication of Journals is no doubt an important feature of the proposed biological society. A plan for this purpose should be worked out to details without which its feasibility cannot be de- termined. Perhaps this could be best done after the society had been organized. It seems to me unwise to base the argument for this new society so largely on the Journal feature, as this alone or as a principal consideration would not suffice to hold it together. The subject of Journals having aroused much responsive interest, the new society should carefully consider the Utility and the financial aspects of this part of the proposition. Howard S. Reed, Virginia Agric. Expt. Station. I am not in favor of the Mathews plan for an Organization of biological soci- eties, because I do not think it would bring about the desired results. It is true that we have many societies, but it is equally true that the societies have been organized and developed to meet definite ends. This is an age of increasing specialization, and individual men can- not begin to cover the field of the biological societies today. Ten 140 Mathews Plan for American Biological Society lOct. years from now the task will be more difficult. Those societies which publish Journals, do so to provide for their own technical papers, that is, to have a place where the work of their colleagues will be segregated from other papers of less professional interest. Most scientists with whom I am acquainted, subscribe personally for two or three of the Journals dealing most closely with their own work, and depend upon the library of their institution for others of general rather than specific interest. The history of the specialized societies shows that there is a constant tendency to break up the older bodies into smaller, more highly specialized groups, if not to form new societies. Where specialized organizations have merged their identity with large societies, there have usually been formed subsequently new special societies to take the place of those which entered the amalgamation. I am heartily in sympathy with Professor Mathews' project to unite the biological interests of the country and to make them more effective in the general development of education, and diffusion of biological knowledge ; and I am in f avor of a scheme of Cooperation or affiliation among societies having allied interests. I would like to see the group of biological societies meet annually at the same time and in the same place, to issue a Joint program and, wherever possible, to hold one or more Joint sessions in which two or more societies might profitably unite, but the ränge of interest is so great that I do not think they could ever be united into one solid Organi- zation. Edward L. Rice, Ohio Wesleyan Univ. There is much which attracts in Prof. Mathews' plan for an Amer. Biolog. Soc'y, with its arrangement for increased distribution and support of our scien- tific Journals. But I am tempted to raise a few questions as to its practicability. 1. If it means "one more" society, is it worth while? Or will enough of our present societies disband to make a place for it? At present we never know which society to attend at any particular time. 2. Isn't Prof. Mathews too optimistic as to the support of the society by the biologists of the country? We are many of us pretty badly strapped financially, and unable to accept many offers which we recognize to be good bargains. 1913] Editoriais 141 3. Woiild the librarles continue to subscribe at old rates for the Journals, or would they depend upon getting them through members of the Society and at the society rate? 4. Would it not be better, if practicable, to keep the dues lower and to include part of the Journals suggested. Few biologists would be vitally interested in more than about half the Journals. This raising of objections on my part does not mean Opposition to the scheme but a desire to see it worked out successfully. I am ready to apply for membership at once. Carl Alois Schwarze, N. J. Agric. Expt. Station, New Bruns- wick. If the various biological and chemical societies could, through unification, organize a biological society along the lines of the Amer. Chem. Soc'y, I believe we could bring order out of chaos. I think many biologists would gladly avail themselves of the opportunity to procure a number of scientific publications at club-rates. E. E. Smith, 50 E. 4ist St., New York City. The value of Mathews' plan for an Amer. Biolog. Soc'y is determined by what American biologists want. If they want exclusive organizations in which membership is recognition of achievement, then the present organizations meet the requirements. If they want a large Organi- zation in which membership is merely recognition of interest and ambition, and whose value is in its strength, then Mathews' plan is an admirable one. That in the main it is practical, it seems to me is demonstrated by similar movements, notably by the history of the Amer. Chem. Soc'y. The argument that it will fail because of the increased expense to the members was put f orward by those opposed to the present Organization of this latter society; but the argument was not supported by subsequent developments. The whole matter is to be decided by whether exclusive membership or strength in numbers is desired. Each has its advantage. Probably the limited membership is more especially advantageous to the individuals ; and strength in Organization, to the science as a whole, since it promotes dissemination. Possibly in time this latter would also react to the advantage of the individuals. Edwin D. Watkins, Univ. of Memphis. The Mathews plan is a splendid one, and would work toward the same end as the Amer. Chem. Soc'y. 142 Mathews Plan for American Biological Society [Oct. R. M. West, Univ. of Minnesota. I have followed the dis- cussion of the Mathews plan for the Organization of an Amer. Biolog. Soc'y with a great deal of interest. While it is based very largely upon the present Organization of the Amer. Chem. Soc'y, it appears to me to differ in one very vital particular. The Amer. Chem. Soc'y Journals have been siiccessful largely through the fact that in the two publications which are issued, the articles are of suffi- cient general interest to encourage chemists in all lines of the science to become members of the society, while Chem. Ahstr. has of course provided a very thorough review of current literature in all branches of chemistry. According to the Mathe ws plan, it apparently is the idea to continue the publication of all or nearly all of the present Journals in biological science. Even with a clubbing arrangement, the subscription to the number of Journals which it would still be necessary to take would be very considerable. If it could be ar- ranged, a much more feasible plan, as it appears to me, would be for a number of the present biological Journals to combine to form a nucleus for the proposed society publication. That some such central Organization is desirable seems indisputable, and I would be heartily in favor of any plan which would tend to put biological science on the same extensive plane that the Amer. Chem. Soc'y is on at present. In the January number of the Biochemical Bulletin, Pro- fessor Mathews outlined in detail a plan for the Organization of an Amer. Biolog. Soc'y. Shortly after its publication, an invitation to The Mathews plan: comment on the plan was forwarded to the mem- A summary of bers of several of the leading biological societies. published opinions ^ sufficient number of replies have been published in the Bulletin^ to reflect very definitely the views of biologists generally on the subject. We have collated, in this summary, ap- provals and objections for the convenience of all concerned in fur- ther consideration of the matter. In Order that the points of view may be shown clearly in their relation to the plan, the essentials of Professor Mathews' sugges- tions are recapitulated on the succeeding pages : 1 Biochemical Bulletin, 1913, ii, pp. 490 and 582 ; iii, p. 134. 1913] Editoriais 143 Plan: Name: The American Biological Society. Objects: (A) To unite the biological interests for the pur- pose of mutual support, education, more effective Cooperation, de- fence and encouragement of research, and to increase the influence of biological knowledge; (B) to start and support a biological ab- stract Journal; (C) to provide new Journals as the need arises; (D) to diminish to members the cost of dues and Journal sub- scriptions. CoNDiTioNS FÜR MEMBERSHip : (a) All membcrs of present biological societies to be eligible; (b) all persons sufficiently inter- ested in biology to pay dues to be eligible; (c) local sections to be established with a certain percentage of the dues returned for sup- port; (d) present biological societies to organize as sections in the general society (membership in a section might be left to the de- cision of the section) ; (e) dues to be sufficient to provide each mem- ber with the abstract Journal and some or all of the biological Journals. Professor Mathews estimates that it would be possible to provide an abstract Journal and thirteen other Journals which cost $83 for a fee of $25 per year. The Journals he listed (in 1908) were the following: Amer. Jour. of PhysioL, Amer. Jour. Anat., Jour. Compar. Neurol., Jour. of Morphol., Jour. Infec. Dis., Jour. Exp. Med., Jour. Med. Res., Biol. Bull., Jour. Biol. Chem., Jour. Exp. Zool., Anat. Rec, Psychol. Rev., and Botan. Gaz. Methods of ORGANIZATION : (a) Either organize the " Natu- ralists " into a new society or form an entirely new society on lines similar to the Amer. Chem. Soc'y; (&) a scale of fees might be pre- sented with an Option as to the Journals desired; (c) the present management of the Journals could be retained and a club rate of subscription offered to members of the society; {d) the Wistar Inst, might be made the Publishing house for the abstract Journal and the other Journals. Objections: The principal ohjections that have been raised to the scheme may be tabulated as follows : I. Doubt of the accuracy of the financial estimate submitted. It is believed that the cost of the thirteen Journals plus that of the abstract Journal would not only exceed Professor Mathews' esti- mate but so much so as to render the plan unfeasible. 144 Mathews Plan for American Biological Society [Oct, 2. Doubt of the financial support of the plan. It is concluded that it woiild be impossible to secure a sufficient number of members to finance the plan without making the dues and subscriptions pro- hibitive. 3. Doubt of the possibility of fusion of the existing biological societies. It is suggested that the present members of these societies would resent any attempt to control their present Organization. 4. Doubt of the desirability of fusing existing biological socie- ties. It is feit that the present specialization is more desirable than fusion. 5. Doubt of the desirability of making membership open to " any one interested in biology." It is assumed that special societies of workers are more desirable and effective than general associations with vague qualification for membership and club rates for the reduction of subscriptions for Journals. 6. Doubt of the desirability of plans that might mean "merely a new society and a new Journal." 7. Doubt of the desirability of a centralization of control for Journals. It is considered that better results are obtained when a Journal is controlled by workers most interested in the particular subject it represents. 8. Doubt of the desirability of a new abstract Journal. It is thought that such a Journal would overlap and duplicate the work of existing foreign abstract Journals. Suggestions. Aside from the doubts presented above, many ofifer constructive criticisms and suggestions to obviate difHculties. Some of these are listed below : A. Suggest a "Federation" similar to that already formed by the Physiological, Biochemical and Pharmacological Societies to control meetings, abstract Journals, etc., and at the same time leave to each society its present autonomy and Journal control. B. Suggest the formation of a business Organization composed of members of the various societies to finance an abstract Journal. This would avoid the necessity of forming a general Organization. C. Suggestions that the division into sections of a general so- ciety be based on allied interests rather than on existing societies. This plan would " concentrate " Journals, both in content and num- I9I3] Editoriais 145 ber, would lower the subscription rates and would reduce the dupli- cation of printed matter. D. Suggestion that the membership and subscription fees be graduated with the idea of allowing a member to elect the expense and Journals he desires. One plan suggested a $10 fee to cover dues, abstract Journal and an election of two other Journals. E. Suggestion that one of the present " Centralblatts " be made the abstract Journal and thus use a medium already established. Addition of .financial support and extension of scope would thus be secured at a minimum expense. Summary of published opinions. In the appended summary are indicated the views of those whose opinions have already been published. Under the heading " Objections," the numbers refer to the tabulated objections given above (page 143). Name Atkinson, Jas. P. Conklin, E. G. Eddy, W. H. Fischer, M. H. Fitz, G. W. Gies, William J. Gortner, R. A. Greene, Chas. W. Hall, Winfield S. Hawk, P. B. Houghton, E. M. Howe, Paul E. Jackson, D. E. Jordan, E. O. Kahn, Max Koch, FC. , Linton, Edwin Lloyd, F. E. McClendon, J. F. McGuigan, Hugh Moore, A. R. Osburn, R. C. Parker, G. H. Schwarze, C. A. Smith, E. E. Stewart, Colin C. Thorndike, E. L. Wiley, Harvey W. Watkins, Edw. D. University or other connection N. Y. City Dep't of Health Princeton Univ. N. Y. High Seh. of Commerce Cincinnati Univ. Peconic, Suflfolk Co., N. Y. Columbia Univ. Carnegie Sta. for Exp. Ev. Univ. of Missouri Northwest. Univ. Med. Seh. Jeflferson Med. Col. Park, Davis & Co., Detr., Mich. Columbia Univ. Washington Univ. Med. Seh. Univ. of Chicago U. S. Bureau of Chem. Univ. of Chicago Washington and Jefson Col. McGill Univ. Cornell Univ. Med. Seh. Northwest. Univ. Med. Seh. Bryn Mawr Col. N. Y. Aquarium Harvard Univ. N. J. Agric. Exp. Sta. 50 E. 4ist St., N. Y. City Dartmouth Col. Columbia Univ. Bur. of Foods a. San., Wash. Univ. of Memphis U. S. Bur. of Anim. Ind. Univ. of California Carnegie Sta. for Exp. Ev. 1 Favors a f ederation. Berg, Wm. N. Burnett, Theo. C. Davenport, C. B. Approval of plan as stated Objections Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Complete None Qualified 61 Qualified I, 2 Qualified 1,8 146 Mathcws Plan for American Biological Society [Oct. University or Approval of Name other connection plan as stated Objections Fetzer, L. W. U. S. Dep't of Agric. Qualified 62 Hargitt, Chas. W. Syracuse Univ. Qualified 3 Henderson, V. E. Toronto Univ. Qualified i Henderson, Yandell Yale Univ. Qualified i, 2 Hepburn, J. S. Food-Research Lab. Qualified 2, 3 Hewlett, A. W. Univ. of Michigan Qualified 4^ Hoskins, R. G. Starling-Ohio Med. Col. Qualjfied 3 Hough, Theodore Univ. of Virginia QuaHfied 3, l Howell, W. H. Johns Hopkins Univ. Qualified 4* Hyde, Ida H. Univ. of Kansas Qualified i, 2 Kingsley, J. F. Tufts Col. Qualified i, 2 Lothrop, A. P. Columbia Univ. Qualified 5 Macleod, J. J. R. Western Reserve Univ. Qualified 2 McNeal, W. J. N. Y. Post-Grad. Med. Col. Qualified 3 Mann, Gustave Tulane Univ. Qualified(Suggestions) Martin, E. G. Harvard Med. Col. Qualified l Morse, Max Univ. of Wis. Qualified 3 Park, Wm. H. N. Y. City Dep't of Health Qualified 3 Pearl, Raymond Maine Agric. Exp. Sta. Qualified 3, 5 Peters, Amos W. Training Seh., Vineland, N. J. Qualified 3 Reighard, Jacob Univ. of Michigan Qualified 2, 7 Rice, Edward L. Ohio Wesleyan Univ. Qualified 2, 6^ Rockwood, E. W, Ohio State Univ. Qualified i, 2, 3 Todd, J. L. McGill Univ. Qualified 3 West, R. M. Univ. of Minnesota. Qualified i, 2 Wood, F. C. Columbia Univ. Qualified 3 Barnhart, J. H. N. Y. Botan. Garden Opposed 2, 6 Bergey, D. H. Univ. of Pennsylvania Opposed 4, 6 Bigelow, R. P. Mass. Inst, of Tech. Opposed 2, 4, 8 Carlson, A. J. Univ. of Chicago Opposed i, 4, 5, 8 Crile, G. W. Western Reserve Univ. Opposed 4 Dahlgren, Ulric Princeton Univ. Opposed 4 Davis, Bradley M. Univ. of Pennsylvania Opposed 2, 4 Dox, Arthur W. Iowa State Col. Exp. Sta. Opposed 4 Gager, C. Stuart Brooklyn Botan. Garden Opposed 4 Hanzlik, Paul J. Western Reserve Univ. Opposed 2, 4, 8 Langworthy, C. F. U. S. Dep't of Agric. Opposed 4 Maxwell, S. S. Univ. of California Opposed 6 Pearce, Richard M. Univ. of Pennsylvania Opposed 4 Reed, Howard S. Virginia Agric Exp. Sta. Opposed 4 Sollmann, Torald Western Reserve Univ. Opposed 4 SUMMARY OF THE VOTE : Total number voting 73 Complete approval 29 Qualified approval 29 Opposed 15 Obj ection i raised by 11 Obj ection 2 raised by 14 Objection 3 raised by 12 2 Approves the abstract-journal plan. 3 Favors a federation. * Favors a society to finance the proposed abstract Journal, s Hopes to see the plan consummated. igi3] Editorials 147 Obj ection 4 raised by 15 Obj ection 5 raised by 3 Obj ection 6 raised by 6 Objection 7 raised by i Obj ection 8 raised by 4 While it is admittedly not permissible to base sweeping conclu- sions upon a vote of only seventy-three individuals, yet these indi- viduals are fully representative of the workers in the biological Sciences, and in general they approve the plan. A Classification of the objections shows that, aside from the doubt regarding the finan- cial estimate and support, the most important objections are those to the fusion of existing biological societies. In fact these are the important objections, for the first two can be established only by a general canvass of the Situation, while the latter threaten the plan in its inception. The question then arises : Can these essential objec- tions be met? The summarized suggestions are instructive in this particular: A federation instead of a general society; a special busi- ness Organization to finance the proposed abstract Journal; auton- omy for the biological societies as the constituent sections. Such suggestions are valuable for the formulation of opinion, but are not decisive until supported by numbers sufficient to afford a working basis for Organization. The views already expressed stimulate thoughts of more effective Organization and should be conducive to that end. The Biochem- ICAL Bulletin suggests that at the next annual meetings of the biological societies, free discussion of this entire matter be included in the order of business of each section and that the results of this discussion, together with the vote of each society on the evolved plan, be formulated by the respective secretaries and sent to the Bulletin for publication in its January issue. The coördinated conclusions might serve as a dynamic basis for the future. Reprints of this summary would be furnished to societies upon request. The Bulletin is anxious to assist in every way to a concrete conclusion in the matter. Walter H. Eddy. The dogmatism of experience is a most dangerous clog to scientific progress. — Dunning. 148 Hormones [Oct. The keenest test of a man comes when he has attained ; the struggle to attain keeps him strong, but the Hne of least Hormones . , . ,- . r^, , resistance soon shows itseli in success. — Black. He that greets Hardship on the threshold of youth may find her a cruel taskmistress, but still a friend. For it is her peculiar func- tion to act as a nurse to the potential conqueror, that he may in the end overcome her and turn her out of doors. Her discipline is rig- orous, but they that in good time show her the door are a hardy breed.— ^/ K. Li. Indifference to the magic of work, the potency of drudgery, is the curse of too many College men. They want to fly before they can creep; they want to be ten thousand dollar men before they are thirty-cent apprentices. Not even College can teach the faculty of absorbing worldly wisdom as a sponge drinks water. Worldly wis- dom is a slow growth. You can't get it in the circus of society or the pantomime of sport; you can't get it in the frivolities of pleasure or the steeplechase of mirth; but you can get it in a man's work among men and nowhere eise. — Glynn. The study of recent literature forces from us the question, why so many students of the (chemical) science, leaving of course the workers in color chemistry and in the synthesis of alkaloids out of account, regard themselves as in duty bound to study the products of the distillation of coal, the relics of a long extinct organic world, and their derivatives, instead of turning their attention to the living World which surrounds them. To invent new methods and to fol- low their application in this region would surely not be less interest- ing than the piling up of many-membered rings. — Lassar-Cohn. The meat of success is savorless without the salt of content. To him that cannot look upon his treasures and the work of his hands and say in his innermost conscience, ''It is good," there is no success. Monumental achievements only madden by their f Utility if they lack the approval of the still, small voice. In the last analysis the human struggle is one for seif -approval. The problem of self-preservation is readily solved by the majority of mankind. It is elemental and comparatively easy. But the problem of winning self-approbation — not the self-approbation of the shallow egotist, but that of the wise, level-headed, introspective person — is elemental and stubbom. He who has solved it is favored of the gods. — Em. Phatic. BOOKS RECEIVED The BiocHEMiCAL Bulletin promptly acknowledges here the receipt of publications presented to it. Reviews are matter-of-fact Statements of the nature and contents of the publications referred to, and are intended solely to guide possible purchasers ; the wishes or expectations of publishers or donors of volumes will be disregarded, if they are incompatible with our convictions re- garding the interests of our colleagues. The sizes of the printed pages are nidicated, in inches, in the appended notices. Untersuchungen über Chlorophyll: Methoden und Ergebnisse. Ey Richard Willstätter and Arthur StoU, Kaiser Wilhelm Inst, für Chemie. Pp. 424 — 7l4 X 4/4 ; M. 20.50. Julius Springer, Berlin, 1913. This comprehensive volume presents unpublished data, obtained by Will- stätter and his pupils in recent years, on the isolation and hydrolysis of chloro- phyl and the Separation and quantitative determination of its component radicals. A complete compilation and revision of the essential data of W'illstätter's clas- sical studies on chlorophyl is included, and the relationship of chlorophyl and hematin is further clarified. The volume is encyclopedic in scope and presents the methods so clearly that it may be used as a laboratory handbook on chloro- phyl. That it will aid and stimulate research on chlorophyl is certain and should be studied by biochemists generally. The volume is beautifully illustrated with eleven plates, which indicate details of the crystalline and spectral characters of the products. The work on which the book is based was a monumental achievement. Gies. The elements of the science of nutrition. By Graham Lusk, pro f. of physiology, Cornell Univ. Med. Col. Second ed. Pp. 402 — 6^ X 3}i ', $3.00 net. W. B. Saunders Co., Phila., 1909. This widely appreciated volume, by a master of the subject in both its theo- retical and practical phases, is one of the best on nutrition. We use it freely in our advanced courses, and await impatiently the appearance of the third edition. Gies. Nutritional physiology. By Percy G. Stiles, assist. prof. of physiology, Simmons Col. ; instr. in physiology and personal hygiene, Mass. Inst, of Tech., Boston. Pp. 271 — 6 X ZV2 ; $1-25 net. W. B. Saunders Co., Phila., 1912. An admirable treatment of nutrition, which is very appropriately dedicated to the author's teacher, Prof. Graham Lusk. The chemical phases of physiology are concisely though none the less effectively considered ; and nutrition is pre- sented from the dynaniic point of view without confusion with food chemistry. A very valuable addition to the growing supply of textbooks in biological chem- istry for beginners. Gies. Essentials of pathological chemistry, including description of the chem- ical methods employed in medical diagnosis. By Victor C. Myers and Morris S. Fine, prof. and instr. in path. chemistry, respectively, at the N. Y. Post-Grad. Med. Seh. and Hosp. Reprinted from the Post-Gradiiate, 1912-13. Pp. 137 — 7 X4; $1-25. Post Graduate (Med. Jour.), N. Y. City, 1913. A very useful compilation of laboratory methods in the pathological chem- istry of digestion and excretion, also of milk and blood, with an appendix of laboratory suggestions. The discussions are practical in guidance and broad in Interpretation. The book is a very handy laboratory manual. We hope the authors will carry it through numerous revisions and extensions, as the science advances and methods multiply. Gies. Books received (con.) Modern research in crganic chemistry. By F. G. Pope. Pp. 324 — 6 X 3% ; $2.25 net. D. Van Nostrand Co., New York, 1913. Restricted, with interesting historical introduction, to chapters successively on polymethylenes; terpenes and camphors; uric acid (piirin) group; alkaloids; relation between color and Constitution of chemical Compounds; salt formation, pseudo-acids and bases; pyrones; ketens, ozonides, triphenyimethyl; and the Grignard reaction. Masterly treatment of each subject. Constitutional formulas used freely and effectively. Gies. An introduction to the chemistry of plant products. By Paul Haas (lecturer on chemistry, Royal Gardens, Kew) and T. G. Hill (reader in vegetable physiology, Univ. of London). Pp. 401 — ^4X7; $2.25 net. Long- mans, Green and Co., 1913. Excellent discussion of the chemistry and biological significance of many of the most important plant constituents. Besides extended treatment of carbo- hydrates, lipins and proteins, chapters are devoted respectively to glucosides, tannins, pigments, nitrogenous bases (alkaloids, ptomaines, purins), colloids and enzymes. Methods of preparation, detection and quantitative determination are numerous and well described. Good subject index. The most valuable recent contribution of its kind to phyto-chemistry. Strongly recommended to biological chemists generally — to botanists in particular. Gies. Practical physiological chemistry. By Sidney W. Cole, demonstrator of physiology, Trinity College, Cambridge. Third edition. Pp. 230 — 4 X 6j^ ; 7s. 6d. net. W. Heflfer & Sons, Ltd., Cambridge, Eng., 1913. Very useful laboratory manual. Subject treated chiefly from static point of view. Practical throughout. Methods well selected. Quantitative pro- cedures given satisfactory attention. Special emphasis laid upon Folin's micro- chemical methods of urinary analysis. Good index. See review by Walter Jones, Jour. Amer. Chem. Soc, 1913, xxxv, p. 1064. Gies. Reagenzien-Verzeichnis enthaltend die gebräuchlichen Reagenzien und Reaktionen, geordnet nach Autorennamen. Dritte Aufl. By E. Merck. Pp. 446—8^ X SYi- Julius Springer, Berlin, 1913. Very useful in a biochemical laboratory. References to original literature with description of each reagent or test. Arrangement favors easy reference to desired author, substance or procedure. Gies. Annual report of the Virginia Polytech. Inst. Agric. Expt. Station for 1911 and 1912. 1913 (13 original papers). Studies from the department of physiology, Cornell Univ. Med. Col., II. 1913. (12 reprints.) Sloane Hospital for Women (N. Y. City) : Obstetrical and gynecological reports. Vol. I, 1913. Edited by Wilbur Ward. 1913. Radium: A m.onthly Journal devoted to the chemistry, physics and therapeutics of radium and other radioactive substances. Vol. I began with issue in April, 1913. Radium Publishing Co., Pittsburgh, Pa. Researches in biochemistry conducted in the Johnston Laboratory, Univ. of Liverpool. Edited by Benjamin Moore, Johnston prof. of biochem, and Owen T. WilHams, demonstrator of biochem. Vol. H; 1908-1911, (27 reprints.) Glycosuria and allied conditions. By P. J. Cammidge. Pp. 467—4X6^; $4.50 net. Longmans, Green & Co., New York; Edward Arnold, London, 1913. The chemical Constitution of the proteins: Part II, Synthesis, etc. 2d ed. (One of the Monographs on Biochemistry.) By R. H. A. Plimmer, Univ. reader and ass't prof. of physiological ehem., University Coli, London. Pp. 107—4^ X 7>4; $1.20 net. Longmans, Green & Co., 1913. OFFICERS OF THE COLUMBIA BIOCHEMICAL DEPARTMENT* Sixteenth year: 1913-14 OFFICIAL REGISTER, SEPTEMBER 31, 1913 William J. Gies: Professor and Executive Officer; Consulting chemist, New York Botanical Garden; Pathological chemist, First Division, Bellevue Hos- pital; Member of the Faculties of N. Y. Teachers College and N. Y. College of Pharmacy. [B.S., Gettysburg College, 1893 and M.S., 1896; Ph.B., Yale University, 1894 and Ph.D., 1897. Instructor, i898-'02; adjunct professor, 1902-05; Professor, 1905-] Paul E. Howe: Assisfant Professor and Secretary of the Staff. [B.S., Univer- sity of Illinois, 1906 ; A.M., 1907 and Ph.D., 1910. Assistant Professor, 1912-.] Alfred P. Lothrop: Associate and Chairman of the Staff. [A.B., Oberlin, 1906 and A.M., 1907; Ph.D., Columbia, 1909. Assistant, i9o8-'o9; instructor, 1909- '12; associate, 1912-.] Emily C. Seaman: Instructor. [B.S., Adelphi College, 1899; A.M., Columbia, 1905 and Ph.D., 1912. Tutor, i909-'io; instructor, 1910-.] Walter H. Eddy: Associate. [B.S., Amherst College, 1898; A.M., Columbia, 1908 and Ph.D., 1909. Assistant, i9o8-'io; associate, 1910-.] Herman O. Mosenthal: Associate; Assistant Attending Physician, Presbyterian Hospital; Assistant Physician, Vanderbilt Clinic; Instructor in medicine. [A.B., Columbia, 1899 and M.D., 1903. Assistant, i9o8-'o9; instructor, 1909- '12; associate, 1912-.] Max Kahn : Associate; Director of the chemical and physiological laboratories, and Consulting physician in dietetics, Beth Israel Hospital. [M.D., Cornell University Medical College, 1910; A.M., Columbia, 191 1 and Ph.D., 1912. Instructor, 1912-13; associate, 1913-] Frederic G. Goodridge: Instructor. [A.B., Harvard University, 1897; M.D., Co- lumbia, 1901. Assistant, 1912-13; instructor, 1913-] Sergius Morgulis: Instructor. [A.M., Columbia, 1907; Ph.D., Harvard Univer- sity, 1910. Instructor, 1913-] Arthur Knudson : Assistant^ 1912-. [A.B., University of Missouri, 1912.] Ethel Wickwire: Assistant, 1912-. [A.B., Tri-State College, 1909-] TuLA L. Harkey : Assistant, 1912-. [A.B., Colorado College, 1909.] William A. Perlzweig: Assistant, 1913-. [A.B., Columbia, 1913-] Christian Seifert: Laboratory assistant, 1898-. Stella Waldeck : Recorder, 1908-. Victor E. Levine: Laboratory assistant, summer session, 1913. [A.B., College of the City of New York, 1909; A.M., Columbia, 1911-] * The work of the department was inaugurated in October, 1898, by Prof. R. H. Chittenden (lecturer and director), Dr. William J. Gies (instructor), Messrs. Alfred N. Richards and Allan C. Eustis (assistants), and Christian Seifert (laboratory assistant). COURSES OFFERED BY THE BIOCHEMICAL DEPARTMENT OF COLUMBIA UNIVERSITY, igiß-'M (Abbreviations: C, Conference; D, demonstration ; L, lecture; Lw, labora- tory work; R, recitation. Odd numbers indicate the first-half, even numbers the second-half, of the academic year; double numerals indicate füll academic year. Courses indicated by numerals in parentheses are not offered during 1913-14) ORGANIC CHEMISTRY 51. Elemextary ORGANIC CHEMISTRY. (Medicol ScJiool.) Introductory to course loi er 102. (Requircd of first year students of mcdicine.) L, D, R, 2 hr. Lw, 6 hr., each section (2). Profs. Gies and Howe, Dr. Lothrop and Messrs. Knudson and Perlzweig. NUTRITION (PHYSIOLOGICAL AND PATHOLOGICAL CHEMISTRY) 61-62. Chemistry of NUTRITION. (School of Phormacy. Reqiiired of can- didates for the Degree of Doctor of Pharmacy.) L, i hr. Prof. Gies. loi or 102. General biological (physiological) chemistry. A course in the Clements of normal nutrition. (Füll course.) Given at the College of Physi- cians and Surgeons, and at Teachers College. College of Physicians and Surgeons — Faculty of Mediane (primarily) : 102 — " Nutrition (physiological chem- istry) 52." Required of first year medical students. (Second half year.) L, R, D, 2 hr. ; Lw, 6 hr., each section (2). Profs. Gies and Howe, Dr. Lothrop and Messrs. Knudson and Perlzweig. (Also given during the last Summer Session by Prof. Gies and Messrs. Perlzweig and Levine.) Faculty of Pure Science (solely) : loi — "Biological chemistry loi." (First half year.) L, R, i hr. ; Lw, 7 hr. Prof. Howe, Dr. Eddy and Messrs. Knudson and Perlzweig. Teachers College — School of Practical Arts: loi or 102 — " Chemistry 51 " and " Household Arts Education 125." L, 2 hr. ; R, l hr., each section (2); Lw, 5 hr., each section (2). (Each half year.) Prof. Gies, Dr. Seaman, and Misses Wickwire and Harkey. (Also given during the last summer Session by Prof. Gies, Dr. Seaman and Miss Harkey.) 201-202. Advanced physiological chemistry, including methods of Re- search IN nutrition. (Pull course. Teachers College, School of Practical Arts.) L, I hr. Lw, 7hr. Dr. Seaman and Miss Harkey. (This course is desig- nated "Household Arts Education 127" in the Teachers College Announcement.) 204. General pathological chemistry. Lectures on nutrition in disease. (Teachers College, School of Practical Arts.) L. i hr. Prof. Gies. (Thiscourse is designated "Chemistry 52" in the Teachers College Announcement.) 2II-212. BiOCHEMICAL methods of RESEARCH, INCLUDING CLINICAL METHODS AND URiNARY ANALYsis IN GENERAL. (Full coursc. Medical School.) L, I hr. Lw, 7 hr. Profs. Gies and Howe, Dr. Eddy, and Messrs. Knudson and Perlzweig. (213-214) BlOCHEMISTRY OF CARBOHYDRATES, LIPINS, PROTEINS AND ENZYMES. (F«// course. Medical School.) L, i hr. Lw, 7 hr. Prof. Gies. 221-222. Nutrition in health. A laboratory course in advanced physio- logical chemistry. (Double course. Medical School.) L, 2hr. Lw, i4hr. Profs. Gies and Howe, and Dr. Morgulis. Courses in Nutrition (continued) 223-224. NuTRiTioN IN DISEASE. A lahoratory course in advanced patholog- ical chemistry. (Double course. Medical School.) L, 2hr. Lw, I4hr, Prof. Gies. 225-226. Nutrition in Disease. {Medical School.) L, i hr, Profs, Gies and Howe, and Drs. Mosenthal and Goodridge. 251-252. Advanced physiological and pathological chemistry, including ALL PHASES OF NUTRITION. (Doiible coursc. Medicol School.) Research. C, i hr. (individual students). Lw, 16 hr. Profs. Gies and Howe, and Dr. Lothrop. TOXICOLOGY 261-262. Effects and detection of poisons, including food preservatives AND ADULTERANTs. (Full coursc. Medicol School.) Lw, 6 hr. Prof. Gies. BOTANY 271-272. Chemical physiology of plants. (Full course. New York Bo' tanical Garden or Medical School, or both.) L, i hr. Lw, 7 hr. Prof. Gies. BACTERIOLOGY (281-282) Chemistry of microorganisms: fermentations, putrefactions and the behavior of enzymes. An introduction to sanitary chemistry. (Full course. Medical School.) L, i hr. Lw, 7 hr. Prof. Gies. SANITATION 291. Sanitary chemistry. (Half course. Teachers College, School of Practical Arts.) L, i hr. Lw, 3 hr. Dr. Seaman and Miss Harkey. (This course is designated " Chemistry 57 " and " Household Arts Education 129 " in the Teachers College Announcement.) BIOCHEMICAL SEMINAR 301-302. BiocHEMicAL Seminar. (Medical School.) 2 hr. Prof. Gies. RESEARCH IN BIOLOGICAL CHEMISTRY Biochemical research may be conducted, by advanced workers, independently or under guidance, in any of the departmental laboratories. LABORATORIES FOR ADVANCED WORK IN BIOCHEMISTRY The laboratories in which the advanced work of the biochemical department is conducted are situated at the College of Physicians and Surgeons, Teachers College, New York Botanical Garden and Bellevue Hospital. Each laboratory is well equipped for research in nutrition and all other phases of biological chemistry. BIOCHEAIICAL LIBRARY Prof. Gies' Hbrary occupies a room adjoining the main biochemical labora- tory at the College of Physicians and Surgeons and is accessible, by appoint- raent, to all past and present workers in the Department. The library contains 2600 volumes and 7000 classified separates. COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION The Biochemical Association holds scientific meetings regularly on the first Fridays in December, February and April, and on the first Monday in June. These meetings are open to all who may be interested in them. SUMMER SCHOOL COURSES See references to courses loi and 102. CONTENTS PAGE A MoDiFiED Hempel Gas Pipette. Stanley R. Benedict i The InFLUENCE OF ArSENIC UPON THE BiOLOGICAL TRANSFOaMATION OF Ni- TROGEN IN SoiLS. /. E. Grcüves 2 The Nature of Humus and its Relation to Plant Life. S. L. Jodidi 17 ClEAVAGE of BeNZOYLALANINE and ACETYLGLYCINE BY MoLD EnzYMES. Arthur W. Dox and W. Eugene Ruth. 23 A CoLOR Reaction of Glycine when Boiled with Chloral Hydrate. Edwin D. IVatkins. 26 Studies on Water Drinking : 15. The Output of fecal bacteria as influenced by the drinking of dis- tilled water at meal time. N. R. Blatherwick and P. B. Hawk. ... 28 A Note on the Determination of Ammonia in Urine. Stanley R. Benedict and Emil Osterberg. 41 Studies of Aeration Methods for the Determination of Ammonium NiTROGEN : 3. The ammonium nitrogen in beef. Jacob Shtdansky and William J. Gies. 45 A Study of the Influence of Cold-storage Temper.'Vtures upon the Chemical Composition and Nutritive Value of Fish. Clayton S. Smith. 54 A Further Study of the Chemical Composition and Nutritive Value of Fish Subjected to Prolonged Periods of Cold Storage. William A. Perlsweig and William J. Gies. 69 The Influence of Chronic Undernutrition on Metabolism. Sergius Morgulis. 72 Nitrogen Metabolism During Chronic Underfeeding and Subsequent ReAlimentation. Sergius Morgulis 74 Proceedings of the Biological Section of the American Chemical So- ciety, Rochester, N. Y., Sept., 1913 : 1. Executive Proceedings. /. K. Phelps, Secretary 76 2. Chairman's Address. Carl L. Aisberg, Chairman 77 3. Scientific Proceedings (Abstracts). I. K. Phelps, Secretary 80 The Biochemical Society, England 96 The Agricultural Colleges and Experiment Stations in the United States. A. C 98 Biochemical Bibliography and Index. William A. Perlzweig 103 Biochemical News, Notes and Comment 112 Editorials: Including additional quotations from letters, and a sum- MARY of PUBLISHED OPINIONS, ON THE MaTHEWS' PLAN FOR THE ORGANI- ZATION OF AN American Biological Society 133 The Biochemical Bulletin is a quarterly biochemical review. It pub- lishes results of original investigations in biological chemistry, miscellaneous items of personal and professional interest to chemical biologists, original con- tributions to research, preliminary reports of investigations, abstracts of papers, addresses, lectures, criticism, reviews, descriptions of new substances, methods and apparatus, practical suggestions, biographical notes, historical summaries, bibli- ographies, quotations, questions, news items, proceedings of societies, personalia, views on current events in chemical biology, etc. Subscription prices. Vol. I: $6.00 (No. i, $1.50; No. 2, $2.50; No. 3, $2.00; No. 4, $1.50). Vol. II: $5.00 (No. 5, $2.00; No. 6, $1.50; No. 7, $2.00; No. 8, $1.00). Vol. III: $2.75 (domestic); $3.00 (foreign.); $5.00 after July i, 1914. Address remittances, manuscripts and correspondence to the Managing Editor, William J. Gies, 437 West sgth St., New York. Vol. III January, 1914 No. 10 Biochemical Bulletin Edited, for the Columbia University Biochemical Association, by tho EDITORIAL COMMITTEE : Herman M. Adler, John S. Adriance, David Alperin, Carl L. Aisberg, D. B. Armstrong, George Baehr, Charles W. Ballard, Louis Baumann, George D. Beal, S. R. Benedict, William N. Berg, Josephine T. Berry, Isabel Bevier, Louis E. Bisch, A. Richard Bliss, Charles F. Bolduan, Samuel Bookman, Sidney Born, William B. Boyd, J. Bronfen Brenner, Jean Broadhurst, Leo Buerger, Gertrude Burlingham, J. G. M. Bullowa, R. Burton-Opitz, A. M. Buswell, R. P. Calvert, A. T. Cameron, Herbert S. Carter, Arthur F. Chace, Ella H. Clark, Ernest D. Clark, Alfred E. Cohn, Burrill B. Crohn, Louis J. Curtman, William D. Cutter, C. A. Darling, William Darrach, Norman E. Ditman, Eugene F. DuBois, James G. Dwyer, Walter H. Eddy, Gustave Egloff, A. D, Emmett. Allan C. Eustis. Benjamin G. Feinberg, Ruth S. Finch, Harry L. Fisher, Kathryn Fisher, Mabel P. FitzGerald, Nellis B. Foster, Fred D. Fromme, C. Stuart Gager, Mary C. de Garmo, Helen Gavin, Mary E. Gearing, George A. Geiger, William J. Gies, Samuel Gitlow, A. J. Goldfarb, H-. D. Goodale, H. B. Goodrich, F. G. Goodridge, Ross A. Gortner, Mark J. Gottlieb, Isidor Greenwald, James C. Greenway, Louise H. Gregory, Abraham Gross, Marston L. Hamlin, Frederic M. Hanes, R. F. Hare, Tula L. Harkey, T. Stuart Hart, E. Newton Harvey, P. B. Hawk, Harold M. Hays, M. Heidelberger, Joseph S. Hepburn, Alfred F. Hess. L. J. Hirschleifer, Benjamin Horowitz, Homer D. House, Paul E. Howe. Secretary, Louis Hussakof, Roscoe R. Hyde, Henry H. Janeway, Max Kahn, John L. Kantor, Edward C. Kendali, J. E. Kirkwood, Israel J. Kligler, Arthur Knudson, Mathilde Koch, Walter M. Kraus, Alfred H. Kropif, Marguerite T. Lee, Victor E. Levine, Charles C. Lieb, B. E. Livingston, Alfred P. Lothrop, Chairman, Daniel R. Lucas, Wm. H. McCastline, Mary G. McCormick, Louise McDanell, Grace MacLeod, C. A. Mathewson, H. A. Mattill, Clarence E. May, Gustave M. Meyer, E. G. Miller, Jr. Sergius Morgulis, Max Morse, H. O. Mosenthal, Hermann J. Muller, Archibald E. Olpp, B. S. Oppenheimer, Raymond C. Osburn, Reuben Ottenberg, A. M. Pappenheimer, Olive G. Patterson, V/. A. Perlzweig, W. H. Peterson, Louis Pine, E. R. Posner, P. W. Punnett, Jessie Moore Rahe, A. N. Richards, Anna E. Richardson, Winifred J. Robinson, Anton R. Rose, Jacob Rosenbloom, William Salant, W. S. Schley, Oscar M. Schloss, H. von W. Schulte, Fred W. Schwartz, C. A. Schwarze, Emily C. Seaman, Fred J. Seaver, A. D. Selby, A. Franklin Shull, C. Hendee Smith, Clayton S. Smith, Edward A. Spitzka, Matthew Steel, Ralph G. Stillman, Charles R. Stockard, Edward C. Stone, Mary E. Sweeny, Arthur W. Thomas, Helen B. Thompson, F. T. Van Buren, Eli^. G. Van Home, Philip Van Ingen, Charles H. Vosburgh, Hardolph Wasteneys, Edwin D. Watkins, William Weinberger, F. S. Weingarten, J. W. Weinstein, Charles Weisman, William H. Welker, C. A. Wells, Harry Wessler, H. L. White, David D, Whitney, Ethel Wickwire, Herbert B. Wilcox, Guy W. Wilson, Louis E. Wise, William H. Woglom, L. L. Woodruff, H. E. Woodward, Hans Zinsser. NEW YORK Bnteredas second-dass matter in the Post Office at Lancaster, Pa. MEMBERS OF THE COLUMBIA UNIVERSITY BIOCHEMICAL ASSOCIATION Honorary Memberr PROF. R. H. CHITTENDEN, First Director of the Columbia Uuiverstty De- partment of Biological (Physiological) Chcmislry and Director of the Sheffield Scientific School of Yale University PROF. HUGO KRONECKER, Director of the Physiological Institute, Uni- versity of Bern, Switzerland PROF. SAMUEL W. LAMBERT, Dean of the Columbia University School of Medicine DR. JACQUES LOEB, Member of the Rockefeiler Institute for Medical Re- search and Head of the Department of Experimental Biology PROF. LAFAYETTE B. MENDEL, Professor of Physiological Chemistry, Sheffield Scientific School, Yale University PROF. ALEXANDER SMITH, Head of the Department of Chemistry, Co- lumbia University Corresponding Members PROF. EMIL ABDERHALDEN, University of Halle, Germany PROF. LEON ASHER, University of Bern, Switzerland PROF. FILIPPO BOTTAZZI, University of Naples, Italy PROF. ROBERT B. GIBSON, University of the Philippines, P. I. PROF. VLADIMIR S. GULEVIC, University of Moscow. Russia PROF. W. D. HALLIBURTON, King's College, London PROF. S. G. HEDIN, University of Upsala, Sweden PROF. FREDERICO LANDOLPH, University of La Plata, Argentina PROF. A. B. MACALLUM, University of Toronto, Canada PROF. D. McCAY, Medical College. Calcutta, India PROF. C. A. PEKELHARING, University of Utrecht, Holland PROF. S. P. L. SÖRENSEN, Carlsberg Laboratory, Copenhagen, Denmark Members Resident in New York City Brooklyn Botanic Garden. — C. Stuart Gager. College of the City of New York. — Wm. B. Boyd, Louis J. Curtman, Benj. G. Feinberg, A. J. Goldfarb. Columbia University: Departments. — Agriculture: O. C. Bowes; Anat- omy: Alfred J. Brown, H. von W. Schulte; Bacteriology: James G. Dwyer, Reuben Ottenberg, Hans Zinsser; Biological Chemistry: Walter H. Eddy, William J. Gies, F. G. Goodridge, Tula L. Harkey, Paul E. Howe, Arthur Knudson, Victor E. Levine, Alfred P. Lothrop, Sergius Morgulis, H. O. Mosen- thal, W. A. Perlzweig, Emily C. Seaman, Chris Seifert, Charles Weisman, Ethel Wickwire; Botany: E. R. Altenburg, C. A. Darling; Cancer Research: W. H. Woglom; Chemistry: A. M. Buswell, R. P. Calvert, Gustave Egloff, H. L. Fisher, P. W. Punnett, A. W. Thomas; Clinical Pathology: Edward Cussler, Peter Irving, Arthur W. Swann; Diseases of Children: Chas. Hendee Smith, Herbert B. Wilcox; Gynecology: Wilbur Ward; Medicine: T. Stuart Hart, I. Ogden Woodruff; Pathology: B. S. Oppenheimer, Alwin M. Pappen- heimer; Pharmacology: Charles C. Lieb; Physiology: Russell Burton-Opitz, Donald Gordon, Leander H. Shearer, Wm. K. Terriberry; Surgery: Hugh Auchincloss, William Darrach, Rolfe Kingsley, Adrian V. S. Lambert, F. T. Van Buren, Jr. ; Therapeutics: Maximilian Schulman; University Physician: Wm. H. McCastline; Vanderbilt Clinic: F. Morris Class, Julius W. Weinstein; Zoology: H. B. Goodrich, John D. Haseman, H. J. Muller, Charles Packard. Colleges. — Barnard: Helene M. Boas, Ruth S. Finch, Louise H. Gregory; College of Pharmacy: Charles W. Ballard; Teachers College: Louis E. Bisch, Members resident in New York (con.) Jean Broadhurst, Ada M. Field, Mary G. McCormick, Mrs. A. P. McGowan, Sadie B. Vanderbilt Advanced studemo. — Graduate {Coli, of P. and S.): Robert Bersohn, Isabel S. Dougherty, Frank R. Eider, H. Gertrude Gates, Hattie L. Heft, Mildred A. Hoge, Jacob Hoffman, Karl J. Holliday, Margaret F. Kelley, Alexander Lowy, Darwin O. Lyon, Edward Plaut, Michael Puorro, G. S. Rosenthal, Fred L. Thompson, W. H. Schliffer, Jr., Arthur P. Tanberg, Jennie A. Walker. — Grad- uate (Teach. Coli): Lucy Gillett, Greta Gray, Madelain Hahn, Hildegarde Knee- land, Helen McClure, Marguerite McLean, Ethel Ronzone, Margaret Stanton, Mary B. Stark. — Medical: Frederick Abramson, M. W. Astarita, Louis Berman, Ernst Boas, David C. Bull, Will H. Chapman, Joseph Felsen, L. H. Ferguson, Wm. Finkelstein, Joseph Goldstone, Julius Gottesman, Martin Holzman, W. S. Horton, Walter F. Hume, H. T. Hyman, Jerome Kohn, Jacob Lattman, J. A. Lazarus, M. V. Miller, F. B. Orr, H. J. Pyle, A. V. Salomon, Isidore Steinman, Harry J. Seiff, Jacob Shulansky, T. F. X. Sullivan, Henry A. Sussman, W. W. Tracey, Grover Tracy, J. I. B, Vail, E. E. Van Derwerker, E. R. Ware, Joseph Yampolsky, F. D. Zeman. [Incomplete.] CoRNELL University Medical College. — Stanley R, Benedict, Robert A. Cooke, Nellis B. Foster, Jessie Moore Rahe, Charles R. Stockard, Geo. W. Vandegrift. EcLECTic Medical College. — David Alperin. FoRDHAM University Medical College. — Benjamin Horowitz. Harriman Research Laboratory. — Isidor Greenwald. Hospitals. — Babies': Morris Stark; Bellevue: Edward C. Brenner, Edward M. Colie, Jr., W. M. Kraus, R. W. Lobenstine; Beth Israel: Charles J. Brim, Max Kahn, Alfred A. Schwartz; City: H. H. Janeway, Louis Pine; Flower: Henry L. Weil; Flushing: Eimer W. Baker; General Memorial: Clinton B, Knapp; Gerntan: H. G. Baumgard, Alfred M. Hellman, Melvin G. Herzfeld, Frederick B. Humphries, Charles H. Sanford, Fred S. Weingarten; Hudson Street: Robert T. Corry; Jewish: Abraham Ravich; Lebanon: Samuel Gitlow, M. J. Gottlieb, William Weinberger; Lutheran: Daniel R. Lucas; Mt. Sinai: George Baehr, Samuel Bookman, Leo Buerger, Burrill B. Crohn, Simon S, Friedman, David J. Kaliski, John L. Kantor, Leo Kessel, Nathan Rosenthal, Harry Wessler; N. Y.: Helen B. Davis, James C. Greenway, Ralph G. Stillman; N. Y. Nursery and Child's: Oscar M. Schloss; Presbyterian: Herbert S. Carter, Russell L. Cecil, Calvin B. Coulter; Roosevelt: J. Buren Sidbury; St. Liike's: Norman E. Ditman, Edward C. Kendall, W. S. Schley. Long Island Medical College. — Matthew Steel. Museum of Natural History. — Louis Hussakof, Israel J. Kligler. N. Y. Aquarium. — Raymond C. Osburn. N. Y. Association for Improving the Condition of the Poor. — Donald B, Armstrong. N. Y. BoTANiCAL Garden. — Fred J. Seaver. N. Y. City Department of Education. — Boys" High School: Frank T. Hughes ; Brooklyn Training School: C. A. Mathewson ; Commercial High School: W. J. Donvan, B. C. Gruenberg, Edgar F. Van Buskirk; DeWitt Clinton High School: Frank M. Wheat; Rastern District High School: Gertrude S. Burling- ham; Girls' High School: Marguerite T. Lee; High School of Commerce: Harvey B. Clough, Fred W. Hartwell; Jamaica High School: Ella A. Holmes, Charles H. Vosburgh; Manual Training High School: Anna Everson; Morris High School: Charles A. Wirth; Newtown High School: Nellie P. Hewins; Wadleigh High School: Helen Gavin, Elsie A. Kupfer, Helen G. Russell, Helen S. Watt. N. Y. City Department of Health.— Charles F. Bolduan, Alfred F. Hess. N. Y. City Normal College. — Beatrix H. Gross. N. Y. Eye and Ear Infirmary.— Harold M. Hays. Members resident in New York (con.) N. Y. Medical College for Women. — Ella H. Clark. N. Y. Milk Committee. — Philip Van Ingen, N. Y. Polyclinic Medical School. — ^Jesse G. M. Bullowa, Mabel C. Little. N. Y. State Food Inspection Laboratory. — Louis J. Hirschlei f er. Post Graduate Medical School. — Arthur F. Chace. Pratt Institute. — Grace MacLeod. Rockefeller Institute. — Alfred E. Cohn, George W. Draper, Frederic M. Hanes, Michael Heidelberger, Gustave M. Meyer, Hardolf Wasteneys. Russell Sage Institute of Pathology. — Eugene F. DuBois. TuRCK Institute. — Anton R. Rose. Vettin School. — Laura l. Mattoon. E. V. Delphey, 400 West S7th Street, Manhattan; Leopold L. Falke, 5316 Thirteenth Avenue, Brooklyn; Mabel P. FitzGerald, 416 East 65th Street, Man- hattan; Abraham Gross, c/o Arbuckle Sugar Co., Brooklyn; Leon M. Herbert, 29 Canal Street, Manhattan; Julius Hyman, 62 East goth Street, Manhattan; Alfred H. Kropff, 619 Kent Avenue, Brooklyn ; Shojiro Kubushiro, 86 Lexington Avenue, Manhattan. Non-Resident Members Agnes Scott College (Decatur, Ga.). — Mary C. de Garmo. Allegheny General Hospital (Pittsburgh). — James P. McKelvy. Carnegie Institution (Cold Spring Harbor, L. I.). — Ross A. Gortner. Drake University Medical School (Des Moines, la.). — E. R. Posner. Forest School (Biltmore, N. C). — Homer D. House, Georgia Experiment Station (Experiment). — C. A. Wells. Indiana Agricultural Exp. Sta. (LaFayette). — Fred D, Fromme. Iowa University Hospital (Iowa City). — Louis Baumann. Isolation Hospital (San Francisco, Cal.). — L. D. Mead. Jefferson Medical College (Phila.). — P. B. Hawk, Edward A, Spitzka. Johns Hopkins University (Baltimore). — ^John Howland, Burton E. Livingston, Edwards A. Park. Kansas State Agricultural College (Manhattan). — Ula M. Dow. MacDonald College (Quebec). — Kathryn Fisher. Mass. Agricultural College (Amherst). — H. D. Goodale. Milwaukee County Agric. School ( Wauwatosa, Wis.) . — Alice H. McKinney. New Hampshire College (Durham). — Helen B. Thompson. New Mexico Agricultltral College (State College). — R. F. Hare. N. J. Agricultural Experiment Station (New Brunswick). — Carl A. Schwarze, Guy West Wilson. N. Dakota Agricultural College (Agricultural College). — H. L. White. North Hudson Hospital (Weehawken, N. J.). — A. E. Olpp. Ohio Agricultural Experiment Station (Wooster). — A. D. Selby. Princeton University. — E. Newton Harvey. Psychopathic Hospital (Boston).— Herman M. Adler. Rensselaer Polytechnic Institute (Troy, N. Y.).— Fred W. Schwartz. Rochester A and M Institute. — Elizabeth G. Van Hörne, Rockford College (Rockford, III.).— Anna M. Connelly. Secondary Scuools.— Brockport State Normal School (N. Y.) : Ida C, Wads- worth; Hermon High School (N. Y.) : Sidney Liebovitz; Indiana State Normal School (Terre Haute): Roscoe R. Hyde; Ingleside School (New Milford, Conn.) : Mary L. Chase; Knox School (Tarrytown, N. Y.) : Clara G. Miller; New Bedford Industrial School (Mass.): Constance C. Hart; North Texas State Normal School (Benton) : Blanche E. ShaflFer; Passaic High School (N. J.) : Hazel Donham, Helene M. Pope; Rochester High School (N. Y.) : David F. Renshaw; State Normal School (Truro, N. S.) : Blanche E. Harris. Non-resident members (con.) Texas A and M College (College Station),— M. K. Thornton. Trinity College (Hartford, Conn.).— Edward C. Stone, R. M. Yergason. TuLANE University (New Orleans, La.)- — Allan C. Eustis. U. S. Department of Agriculture (Wash.).— Carl L. Aisberg, W. N. Berg, H. E. Buchbinder, Ernest D. Clark, George A. Geiger, William Salant, Clayton S. Smith. U. S. Food and Drug Inspection Laboratory (Phila.) .— Harold E. Woodward. U. S. Food-Research Laboratory (Phila.). — Joseph S. Hepburn. University of Alabama Medical School (Birmingham). — Richard A. Bliss. University of California (Berkeley). — William T. Home. University of Chicago. — Mathilde Koch. University of Georgia Medical School (Atlanta). — ^William D. Cutter. University of Illinois (Urbana). — George D. Beal, Isabel Bevier, A. D. Emmett; College of Medicine (Chicago).— Edgar G. Miller, Jr., William H. Welker. University of Indiana (Bloomington). — Clarence E. May. University of Kentucky (Louisville). — Mary E. Sweeny. University of Manitoba (Winnipeg, Can.). — A. T. Cameron. University of Michigan (Ann Arbor). — A. Franklin Shull. University of Minnesota (Minneapolis). — Josephine T. Berry, Louise McDanell. University of Missouri (Columbia). — Louis E. Wise. University of Montana (Missoula). — J. E, Kirkwood. University of Pennsylvania (Phila.). — A. N. Richards. University of Porto Rico (Rio Piedras). — L. A. Robinson. University of Tennessee (Memphis). — Edwin D. Watkins. University of Texas (Austin). — Mary E. Gearing, Anna E. Richardson. University of Toronto (Canada). — Olive G. Patterson, University of Utah (Salt Lake City). — H. A. Mattill. University of Washington (Seattle). — Elizabeth Rothermel. University of Wisconsin (Madison). — Max Morse, W. H. Petersen. Vassar College (Poughkeepsie, N. Y.)— Cora J. Beckwith, Winifred J. Robinson. Wesleyan University (Middletown, Conn.). — David D. Whitney. West Pennsylvania Hospital (Pittsburgh).— J. Bronfen Brenner, Jacob Rosenbloom. Williams College (Williamstown, Mass.). — John S. Adriance. Yale University (New Haven, Conn.). — Lorande Loss Woodruff. Albert H. Allen, Saranac Lake, N. Y. ; Sidney Born, Lemp Brewing Co., St. Louis, Mo.; Emma A. Buehler, Newark, N. J.; Edward G. Gn^», Albany, N. Y. ; Marston L. Hamlin, Yonkers, N. Y. ; F. C. Hinkel, Utica, N. Y. ; Cavalier H. Joüet, Roselle, N. J.; Emile F. Krapf, Standard Chemical Co., Pittsburgh, Pa.; Adeline H. Rowland, Pittsburgh, Pa. ; H. J. Spencer, Factoryville, Pa. ; William A. Talta- vall, Redlands, Cal.; David C. Twichell, Saranac Lake, N. Y.; Isabel Wheeler, Toledo, Ohio. ANNOUNCEMENT. Professional Assistance Offered to Biological Chemists The members of the Columbia University Biochemical Association will coöperate confidentially with any one who desires the Services of biological chemists or who seeks a position in biological chemistry. Address inquiries to William J. Gies, 437 West sgth St., New York. EDITORS OF THE BIOCHEMICAL BULLETIN The editorial committee with the coUaboration of the members and the SPECIAL CONTRIBUTORS: DR. JOHN AUER, Rockefeiler Institute for Medical Research PROF. WILDER D. BANCROFT, Cornell University, Ithaca MR. N. R. BLATHERWICK, Yale University, New Haven, Conn. MR. H. C. BIDDLE, Urhana, III. DR. WALTER L. GROLL, Elisabeth Steel Magee Hospital, Pittsburgh, Pa. DR. CHARLES A. DOREMUS, 55 IV. 52d St., New York City DR. ARTHUR W. DOX, Iowa State College Agric. Experiment Station, Arnes PROF. JOSEPH ERLANGER, Washington Univ. Medical School, St. Louis DR. EPHRAIM M. EWING, N. Y. Univ. and Bellevue Hosp. Med. College DR. LEWIS W. FETZER, U. S. Dep't of Agriculture, Washington, D. C. PROF. MARTIN H. FISCHER, University of Cincinnati DR. MARY LOUISE FOSTER, Smith College, 'Northampton, Mass. PROF. J. E. GREAVES, Utah Agricultural College, Logan DR. V. J. HARDING, McGill University, Montreal, Canada DR. R. H. M. HARDISTY, McGill University, Montreal, Canada DR. J. A. HARRIS, Carnegie Sta. for Exp. Evolution, Cold Spring Harbor, L. I. DR. K. A. HASSELBALCH, Einsen Institute, Copenhagen, Denmark PROF. G. O. HIGLEY, Ohio Wesleyan University, Delaware DR. S. L. JODIDI, U. S. Dep't of Agriculture, Washington, D. C. DR. VERNON K. KRIEBLE, McGill University, Montreal, Canada PROF. FRANCIS E. LLOYD, McGill University, Montreal, Canada PROF. JOHN A. MANDEL, N. Y. Univ. and Bellevue Hospital Med. College PROF. WILLIAM MANSFIELD, College of Pharmacy, Columbia University PROF. ALBERT P. MATHEWS, University of Chicago PROF. SHINNOSUKE MATSUNAGA, University of Tokyo, Japan PROF. VICTOR C. MYERS, N. Y. Post-Graduate Med. School and Hospital DR. RAY E. NEIDIG, Iowa State College Agric. Experiment Station, Arnes DR. THOMAS B. OSBORNE, Conn. Agric. Experiment Station, New Haven MR. EMIL OSTERBERG, Cornell University Medical College, New York City DR. AMOS W. PETERS, The Training School, Vineland, N. J. DR. I. K. PHELPS, U. S. Dep't of Agriculture, Washington, D. C. PROF. R. H. A. FLIMMER, University College, London DR. W. EUGENE RUTH, Iowa State College Agric. Experiment Station, Arnes PROF. R. F. RUTTAN, McGill University, Montreal, Canada DR. JESSE A. SANDERS, University of Indiana, Bloomington DR. E. E. SMITH, 50 East 4ist St., New York City DR. A. E. SPAAR, City Hospital, Trincomalee, Ceylon PROF. UMETARO SUZUKI, University of Tokyo, Japan DR. :CLARENCE J. WEST, Rockef eller Institute for Medical Research MISS ANNA W. WILLIAMS, State Agric. Coli, Manhattan, Kan. PROF. E. WINTERSTEIN, Polytechnic Institute, Zürich, Switzerland DR. JULES WOLFE, Pasteur Institute, Paris ^^y^^--3-— >" .— w-^ / ^^'-— \^ Z-^m^^ — -m^yl^-m^ BiocHEMiCAL Bulletin Volume III JANUARY, 19 14 No. 10 DINNER TO HENRY HURD RUSBY The Alumni Association of the College of Pharmacy of the City of New York honors Dean Rusby The eighteenth annual dinner of the Alumni Association of the N. Y. College of Pharmacy was given at the Chemists' Club, New York, on December 17, 19 13, in commemoration of the twenty- fifth anniversary of Dean Rusby's appointment to the faculty of the College. The dining room was filled to its capacity with a happy Company of alumni and other friends of Doctor Rusby. Toward the close of the dinner Dr. Joseph Weinstein, president of the Alumni Association, addressed the gathering, saying: "It is with a feeling of pleasure that I remind the diners that we have assembled to celebrate the silver jubilee of our beloved dean, Henry H. Rusby. I call upon all present to join with me in paying a tribute to and in felicitating the dean." Dr. Weinstein then intro- duced Professor Curt P. Wimmer, the chairman of the dinner com- mittee, as the toastmaster. Dr. Wimmer said that no one " outside the f amily " had been invited to speak on this occasion, because the dinner committee feit that everything should go the " Rusby way," which is a simple way, but a way of results and achievements. After reading a number of letters from distinguished pharma- cists who were unable to be present, the toastmaster introduced Professor William H. Carpenter, Provost of Columbia University, who said in part: "I feel profoundly grateful that the chairman has classed me as a member of the great and harmonious family of the College of Pharmacy. I was somewhat shocked when I received the invita- tion to attend this dinner because it was stated to be the twenty-fifth 149 LIRRARV NEW YOki 80TAN1CAI QAKUI3N. 150 Dinner to Henry Hurd Rusby [Jan. anniversary of Dean Rusby's connection with the College of Phar- macy. I can not think of Dean Rusby as of an age that a twenty- fifth anniversary would seem to indicate, for he has none of the characteristics that we are apt to regard as the characteristics of age. In clearness of vision, in mind and in heart he is young and he will always remain young no matter what the measure of his years may be. "I came here to-night to represent the University and to lay my tribute at this twenty-fifth milestone which marks the broad highway of the Dean's progress through life. I value my associa- tion with Dr. Rusby. I know of his scientific accomplishments, of the depth and the breadth of his influence in that great field in which he is a laborer. We of Columbia value our connection with the College of Pharmacy as one of the great and important parts of the University. The College of Pharmacy has grown in size and in influence on account of the realities of its purposes and its achievements and on account, too, of the wise administration of its dean. "Dr. Rusby is a 'bonny fighter.' He has fought a sturdy, valiant and honest fight for higher educational Standards in phar- macy. He has contributed in no small part to make the pharmacist more and more a powerful influence for good in the Community. He has helped to make the profession of pharmacy one of the im- portant members in the whole group of the learned professions. Happy is the university that has such a man among its teachers! Thrice happy is that department of our University to have as its leader a man of such wide horizon, such high ideals, such steadi- ness of purpose and serenity of soul. May Dean Rusby live long and may he enjoy the deserved fruits of his labors." Dr. Charles F. Chandler then addressed the association : " Dr. Rusby is one of my boys. I don't remember him as one of my stu- dents, since I knew intimately only the tail-enders of the class. Professor Rusby should be a happy man — first, because he has been a teacher all his life; second, because he is sympathetic with and interested in his students. My first real acquaintance with Dr. Rusby began when he joined the faculty of the College of Phar- macy. When Dean Rusby lectures to the students he impresses them with the f act that he knows wPiat he is talking about. This is igi4] William Mansfield 151 a great asset to a teacher. I don't think he has a superior, and I am sure he has few equals as a teacher. Since Dean Rusby has been connected with the College it has prospered. Each year new courses have been added, new members added to the f aculty, and new facilities provided for the students. This has resulted in an in- creased attendance. In conclusion, I hope Professor Rusby will be here to celebrate his half -Century with the College." Dr. William J. Schieffelin said : " It is an anti-climax for me to follow Professor Chandler. When I returned from Munich in 1889 I knew Professor Rusby as a slender, curly-haired young man. We, the trustees, are always able to bank on Professor Rusby. In Order to secure pure drugs he penetrated the wilds of South America and Mexico. In later years he has continued the fight for pure drugs. It is great to feel that we have scientific men who care for truth. Dr. Rusby has stood like a rock for the highest Standard of purity in drugs. I am proud to have been connected with him, and with the trustees of the College of Pharmacy. Dr. George C. Diekman said in part : " It will be impossible for me to say one-half of that which I have in mind to say concerning the one we have come here to honor this evening, and who com- pletes this year twenty-five years of uninterrupted, capable, honor- able and faithful service to our College. How efficient these Serv- ices have been, and how faithfully they have been performed, is not only known to every member of our f aculty, but to everyone connected with the College as well. It is really my good fortune to have the opportunity of addressing those here assembled concerning one about whom so much that is good can with truth be said. Pro- fessor Rusby is so well known to all present that my further re- marks concerning him will, I am certain, be endorsed in every detail. " During my long and at all times pleasant and cordial associa- tion with Dean Rusby two qualities of the man have impressed themselves upon me more than any others. They are honesty of purpose and unselfishness. I need say nothing here concerning his many other admirable qualities, nor about his reputation as a scien- tist and teacher. A correct estimate of the two qualities referred to can only be made in case of any man, after long-continued asso- 152 Dinner to Henry Hurd Rusby [Jan. ciation aiid study and Observation at close ränge, and such have been my opportunities in this case. " I know of no one who is more honest of purpose than is Pro- fessor Rusby. Nothing can induce him to withdraw when once he has decided to lend his influence to any cause. He has on many occasions entered into a contest against long-estabHshed customs and abuses, or a contest against individuals, when it would have been greatly to his advantage to have remained neutral, or at least to have fought less vigorously and persistentl3^ In many Hke situ- ations a man less honest in purpose would have withdrawn from the contest when once it became clearly apparent that in gaining victory in a contest for principle he was to be personally a loser. Dean Rusby could not, even if he desired, say one thing and do another. Once he has satisfied himself that he is right he proceeds to act regardless of his own personal interests. "And now, Dean Rusby, I come to the most pleasant part of that which I am privileged to do here to-night. You already are aware of the high esteem in which you are held by the members of the faculty. Nothing can add or take from that esteem. And in Order that you may be reminded, if indeed a reminder is necessary, of the very cordial and f raternal relations which exist between your- self and the members of the faculty (and I am using the term faculty in its broadest sense), when you are at home and away from the scene of your labors, the members of the faculty decided to present you with a tangible evidence of their love and affection in the form of the loving cup I now have the honor to place in your hands. "We feel and know that you will prize the sentiment which prompts and accompanies the gift far more highly than its intrinsic value. We ask you to accept it as a token of fidelity and loyalty, which will endure as long as we are permitted to labor together for the honor and glory of our grand old institution, the College of Pharmacy of the City of New York. God bless you." Dr. Rusby said, in accepting the loving cup : "As I sat and listened to the different Speakers I feit that I had much to say. But after receiving this beautiful gift, words go from me. It is difficult to reply. As I have listened to the different things said 1914] William Mansfield 153 about me I have pinched myself to see if I were alive. Such pleas- ant things are usually said about a man after he is dead. You have all heard what I have done for the College. What has this College done for me ? It has moulded and built my soul. I have profited by my association with the other members of the faculty and with the trustees. I have grown in character each year of my associa- tion with the College. In my work as dean of the College I have been brought closely in contact with the worst Clements of the various classes. Instead of removing these men from the College I have admonished them and saved them from themselves. The College of Pharmacy is not a one-man College; it is a College of growth. It is a College which always does what it says it will do. The spirit of harmony is the dominant note in the College of Pharmacy." Mr. Thomas Main, in speaking for the alumni association, said in part : " I was one of the signers of the petition to form an alumni association. The other signers were : P. W. Bedford, class of 1858; D. C. Robbins, class of 1836; William Hegeman. class of 1837; E. L. Milhau, class of 1856; C. B. Smith, class of 1863; Theobald Frohwein, class of 1863; J. W. Ballard, class of 1870; Edwin Henes, class of 1871. The call was sent out on May 10, 1871. Six of the nine signers have now passed away. Those left are John W. Ballard, who, having had a successful drug störe in Davenport, la., for many years, is now a banker; his drug störe is now conducted by one of his sons under the name of the Ballard Drug and Dental Company. The other two survivors are Edwin Henes, of this city, and myself. P. W. Bedford played a most important part in forming the association. He not only signed the call but by his influence obtained the names and secured the influ- ence of the signers, and I strongly suspect was instrumental in in- fluencing D. C. Robbins, of the class of 1836, who was then con- sidered the Nestor of the wholesale drug trade not only of New York, but also of the entire United States. He accepted the presi- dency of the new association for the first two years of its existence, which insured its success from the start. " Mr. President, I venture to think that one of the things the Alumni Association may well be proud of is the part it took in 154 Dinner to Henry Hurd Rusby [Jan. introducing our dear friend and guest of the evening, Dr. H. H. Rusby, to its members and the members of the College. Dr. Rusby, during 1885 and 1886, had been engaged in an exploring and botan- izing expedition which led him from the western coast of South America across the Andes and from the sources of the great river Amazon to the Atlantic Ocean. In February, 1888, he was intro- duced to an audience composed of members of this Association, members of the College and to other scientific societies, by Chas. F. Heebner, then president of this Alumni Association, when Dr. Rusby gave a graphic account of his journey with its scientific results. Dr. Rusby's lecture created a profound impression at that time and I believe resulted in his appointment to the Chair of Materia Medica and Botany in the following year. " With the advent of Dr. Rusby as Professor in our College came his 'object lesson' methods of teaching his subjects, which have been so successful as to compel a complete revolution from former methods of teaching materia medica, not only in our own Institution, but in all other Schools of Pharmacy that consider themselves up-to-date. "The year 1914 will mark the 25th year of Dr. Rusby's con- nection with our Institution. He is now our senior Professor in active service, and Dean. During all these years he has given his best energies and his best thought to his work in the College, and has been a most important factor in placing and maintaining the College of Pharmacy of the City of New York in the proud Posi- tion it holds to-day among the schools of pharmacy of the United States and of the world. " As the first lecture of Dr. Rusby in our College was delivered upon the invitation of the Alumni Association, it seems eminently proper that this Association should be the first to congratulate him upon the near completion of his twenty-five years of important Serv- ice in the College. Mr. Dean, you are here to-night as our hon- ored guest, and it is my great privilege on behalf of the Alumni Association to present you with this silver set, tendered as an appre- ciation of your long, honorable and valued service to our beloved alma mater. During this time you have done your duty as you saw it, without fear or favor, and have earned the friendship and high esteem of all our members." 1914] William Mansfield 155 In conclusion, the toastmaster expressed the hope that the dinner had served to show the Dean the love and affection which his col- leagues and old students feel for him, and to voice the respect and esteem in which he and his work are held by all who know him. William Mansfield New York College of Pharmacy, Columbia University VIEWPOINTS IN THE STUDY OF GROWTRi LAFAYETTE B. MENDEL From whatever Standpoint the Student of science considers liv- ing organisms he soon learns that there are certain fundamental characteristics peculiar to their protoplasm and distinguishing it from what is commonly termed unorganized matter. Life without growth and activity, without the power of " automatic development, self-preservation, and reproduction," is at present inconceivable. Whether or not the artificial production of hving matter from that which is hfeless may some day be accomplished need not concern us now. As Karl Pearson has written: "There is mystery enough here, only let us clearly distinguish it from ignorance within the field of possible knowledge. The one is impenetrable, the other we are daily subduing." I have termed the subject of this review " viewpoints " to empha- size what is all too frequently forgotten, namely, that the data of science are continually changing as the result of the addition of new facts and the modification of the known. Such changes bring with them new sequences — they compel us to new conclusions and sug- gest new inquiries. Donaldson recently remarked that : " The atti- tude toward knowledge during our student days is almost neces- sarily such as to throw the idea of change into the background and unduly emphasize the permanency of the things taught. The facts are otherwise" (Science, July 25, 1913, p. 109.) Furthermore, it is quite as important for the scientific investigator to formulate clearly the problems which his experience dictates as to be con- cerned solely with his experimental researches. (I confess, how- ever, that I often feel sympathy with a remark attributed to the late Professor Atwater, that his idea of a scientific man's heaven 1 This paper was read in part at the third annual dinner of the Columbia University Biochemical Association in New York City, November 21, 1913; sub- sequently, also, before the local chapters of the Society of the Sigma Xi at the University of Kansas, the University of Missouri, and Washington University, St. Louis. The charts and photographs are not reproduced here. 156 1914] Lafayette B. Mendel 157 was a place where one could work incessantly without the necessity of preparing one's results for publication. ) It is surprising that in comparison with other topics of physio- logical study so little has been published in the past about growth ; and much of what has been written invades the domain of hypoth- esis and speculation. Together with inheritance, growth provides for the permanence of the various external manifestations of hfe. The Problems of agriculture — the production of the plant products from the soil and animal husbandry — are based upon considerations of growth. I need scarcely add that it would be of immense direct importance to man and to medicine to know more about " the regu- latory power which presides over growth," so that we might derive more practical applications of the discoveries in this field. In ulti- mate analysis all of our material prosperity, indeed, the very pos- sibility of the maintenance of the race, depends upon the manifesta- tion of growth. The dominating viewpoint in the consideration of the phe- nomena of growth will be determined in large degree by the train- ing and immediate interest of those engaged in the analysis thereof. Some investigators — and these represent by far the largest group — have directed their attention to the purely morphological aspects of the subject. Cytological phenomena, the changes in the number and size of the body cells, the interrelation of the tissue components, the comparative proportions of nucleus and cytoplasm, and similar structural differentiations have engrossed their interest. Others, again, have encompassed the problems of growth from what may be called a dynainic Standpoint. By these, developmental processes are looked upon as expressions of changes in equilibrium. Chemical and physico-chemical reactions are brought into play. The language of a chemist is exemplified in the contention that growth appears to be "the expression of autocatalytic chemical reaction," and par- ticular cycles of growth of an organism are accordingly shown to obey a precise mathematical formula. Somewhat more tangible than the distinctly hypothetical analyses of the chemical character of life phenomena and the physical structure of living matter are the conceptions of those scientists who aim to correlate growth with nutrition, emphasizing in some cases the material side and express- ing the relationships in units of matter, or in others combining with 158 Viewpoints in the Study of Growth [Jan. this the energy aspects and speaking in terms of calories. Lastly there are the considerations of the gross increase in size and form and other features, in which Statistical factors assume the dominat- ing importance. None of these aspects are without significance in the conception of growth; and if the biology of its processes still seems obscure, we must remember the complexity of the factors and forces which interplay in life. What is growth? One cannot penetrate far into the literature of the subject without meeting with a bewildering confusion in the significance of the term. Various expressions, such as growth, organic growth, development, and euplasia have been applied to the same phenomena ; and the numerous attempts to define their mean- ing and precise application have almost invariably ended in a failure to meet criticisms which might be urged against each definition sub- mitted. The most general definition of growth is " increase in volume " or " increase in size " (Huxley) . It has been pointed out, however, that increase in volume does not always serve as an index of organic growth ; f or the increase may merely be due to swelling. Sachs defined growth as an increase in volume accompanied by a change in form. Little is gained by defining growth as the result of a process of molecular intussusception (Huxley) or as due to excess of assimilation over disassimilation (Verworn). No uni- versally acceptable definition has been framed; nor is it likely that one can consider all of the manifold features of growth in a single category. Some of the more familiär uses of the word growth are even more confusing than the so-called scientific definitions. Thus it is Said that a tumor " grows," though the processes described may be quite unlike the growth of an organism. And in the " growth " of a hydrocephalus the analogy is stretched still further. What is needed today is less of theory and more of facts upon which to build a more substantial conception of growth and formulate its fixed characteristics in words. The justification to dwell for a moment upon a review of the more adequate definitions proposed lies in the emphasis which this may throw upon some of the problems encompassed in the concept of growth. Lee has presented the analysis of growth in these words : " All growth, whether of the cells, the tissues, or the organs, is the result of no more than three processes, viz., multiplication of 1914] Lafayette B. Mendel 159 cells, enlargement of cells, and deposition of intercellular substance, the first two being the most potent of all. Increase in the number of cells is largely, although not wholly, an embryonic phenomenon ; increase in the size of cells and deposition of intercellular substance are especially important from the later embryonic period through the time of birth and up to the cessation of body-growth. The most obvious result of the growth of the cells, the tissues, and the Organs, is growth or increase in size of the body." If organisms are composed of both living matter and formed substance, it is evident that a growth may result from the increase of volume of either of these. This viewpoint is somewhat broader than that which governs the usual attitude of the physiologist. If body weight is taken as the measure of growth, it may be pro- nouncedly altered by deposition or removal of reserve materials, such as glycogen and fat, as well as by the retention of water and other products. In the narrower and stricter sense this is not growth. In many individuals the end of growth, as the physiolo- gist intends to apply the term, is reached long before the gain in body weight ceases to manifest itself. Real growth in man, for example, usually is considered to stop at an age of 25 to 30 years when body length has reached its maximum. Many individuals continue to gain in weight and size until they are 50 years old ; but this is of quite a different order than the gains in earlier years. All of these considerations — cell division, increase in number or volume of cells, etc. — fail to take into consideration some of the most striking features of growth as it applies to the higher organ- isms. Perfect growth and development implies a far reaching cor- relation of the various parts of the body. An upset in this nicely balanced relationship is speedily recognized as an anomaly. Energy and matter are insufficient to explain the consummation and main- tenance of a normal as contrasted with an abnormal composition of the cells. The specificity of growth is something marked, particu- larly when we contrast the normal with perverted growth. The regularity and characteristic individuality of the ontogenetic devel- opment Seen in each species has found expression in the explicit defi- nition by Schloss : "So können wir das normale Wachstum eines jeden Organismus auffassen als artspezifische korrelative Vermehr- ung der Körpermasse in bestimmten Zeitabschnitten." The corre- i6o Viewpoints in the Study of Growth [Jan. lation featiire in respect to composition and form, the time relations, the anomalies and irregiilarities involved, are thus brought out. Here, as elsewhere in physiology, abnormalities have often given the clue to the iinderstanding of normal processes. Inasmuch as growth involves a more or less continuous change, there is need of some criterion thereof — some suitable method of ascertaining and measuring it. This is by no means as readily ac- comphshed as might appear at first glance. Changes in body weight are not always reliable guides. There are increments of weight which are transitory and those which are permanent. Uncorrelated increments of mass may occur in the body long after the conclusion of what is properly termed the period of growth. A gain of weight at the middle life, for example, may be due to a deposition of re- serve materials. Increase in stature and certain other dimensions are not without significance as indications of growth; for they allow US to estimate the correlation between weight and size. Without suitable proportionality of form perfect growth, to say the least, cannot go on. How illusory the dependence on any one criterion of growth, such as body weight, may be is illustrated by the interesting studies of Waters and of Aron. These investigators have described condi- tions of adverse nutrition in young animals in which no change '^in body weight occurred during long periods of time. Nevertheless the animals " grew " f rom the Standpoint of changes in stature and body proportions. In some cases the skeleton grew at the expense of other parts of the body, especially the flesh. Many of the organs retain their weight under such conditions, while the brain grows to reach its normal size. Is the constancy of live weight an indication of lack of growth? If one allows the term "growth" for the phenomena of distor- tionate change here cited, it must be admitted with Aron that the " growth " depends principally on the tendency to grow possessed by the skeleton. Evidently, then, there are limitations to the de- pendence on weight measurements. They teil us nothing about the intricate changes in the tissues that may distinguish the adolescent from the adult form, nor do they serve as an index to the desired development of the body in three dimensions, — in other words of proportionate growth. Nevertheless the study of growth by weigh- 1914] Lafayette B. Mendel 161 ing has hitherto furnished the best measure of the total changes of the body. During growth the Compounds and elements of the body are undergoing change and chemical variations. A detailed knowledge of what these are is desirable from many points of view. At the present day only the beginnings in this direction have been made. The work of obtaining statistics as to the varying chemical compo- sition of the component parts of the growing organism at different periods of development involves laborious analyses, with the added difficulty that in order to secure data an experiment must be stopped by the sacrifice of the subject for examination. The correlation in development between different organs and tissues, the appropriate symmetry and interrelation of the parts, the changes in the " körper- gestaltende " as well as the " körpervermehrende " f unction are de- serving of close investigation. The inroad into these and related fields has been begun. When the statistics derived from periodic measurements of the continuous changes going on in young animals are expressed in graphic form, the so-called curves of growth are obtained. The fixity of the growth curve for the different species and, in so far as this point has been studied, of the sexes in each species is perhaps the most remarkable phenomenon of growth. " We are aware of no conditions compatible with life in which the general character of the growth curve with its acceleration during adolescence can be altered. Minor variations may, however, arise." Variations in the growth of dififerent individuals are for the most part inborn — inherited fundamental characteristics of the individual. We know of no method or means of altering the peculiarities of growth. Nutrition, which is often looked upon as a Controlling factor, can do no more than give free scope to the inherent tendency to grow. As Rubner has remarked, the hope of altering this tendency is quite as utopian as the attempt to prolong life indefinitely. It has already been pointed out that the cells need not be the sole factors involved in the increase of size attending growth. Extra- cellular parts may come into play. Nevertheless the importance of the cells as the material basis of biological phenomena and the seat of physiological functions has tended to center attention upon these histological units in growth. We are brought face to face with the 102 Viewpoints in the Study of Growth [Jan. question whether the size of an organism depends upon the size of its component parts — its cells, — or upon their number. Does an organism grow by increase in the number of new cells or by an enlargement of the old ones ? In other words, is there a relation be- tween body size and cell size? It would take us too far afield to examine in detail the evidence bearing on this and related morpho- logical questions of the nature of growth. They have been vigor- ously debated. The answers probably differ for unlike species and certainly for different tissues. If in the lower forms of life differ- ences in ultimate body size are due in the main to differences in cell number, the cell size being approximately constant, it by no means' follows that this Statement holds throughout the progressive stages of growth in higher organisms where other factors than cellular increase and enlargement also are involved. Perhaps we may dis- tinguish between the processes of tissue dififerentiation and growth by reference to the changes in the number and the size of the cells respectively. Much has been made of the necessity of maintaining a certain proportion between the size of the nucleus and that of the cytoplasm composing the body of the cell. Doubtless physiological functions are closely associated with the presence of the nucleus and it appears that when a cell is to attain a very large size it is almost always found to contain several nuclei. Indeed, as we shall see, Minot has presented the relative increase in the cytoplasm with accompanying differentiation as the proximate cause of senescence. Experience shows that there is a fairly definite upper limit in size which the individuals of any species rarely exceed. There are forms for which the variations may be very wide (as has already been pointed out) ; and it is reported that some of the lower forms, e. g., actinians, can be caused by suitable feeding to reach a colossal size far beyond that which they ordinarily attain in nature. To the mammalian species with which we are primarily concerned here, however, this does not apply except in the limited degree determined by heredity. Why the body size is thus fixed is not known. The fact of its invariable character makes it possible to apply quantita- tive methods to the study of growth with some confidence in the consequences which are to be expected from any normal procedure, and with some appreciation of the Standard to which proper growth should conform and by which all deviations are to be measnred. 1914] Lafayette B. Mendel 163 Quite aside from the question as to what initiates growth, the capacity to grow — the " Wachstums fähigkeit," " Wachstumsmög- Hchkeit" as it has variously beert termed — is commonly made a property of the cells of the organism. Whatever may be the ulti- mate cause of growth, the capacity to grow is currently associated with a youthful character of the cells involved. From this stand- point age is an important factor in the possibility of growth. • An embryonic condition of the cells is accordingly most favorable to growth. Minot's third law of age, dealing with the growth func- tion reads : "The rate of growth depends on the degree of senes- cence." From a purely theoretical Standpoint it is quite conceiv- able that ordinary cessation of growth may be due to a natural in- hibitory factor which develops in the course of time rather than because the capacity to grow is lost. Nevertheless the idea that the growth power inevitably declines and is lost with age has found a firm foothold in physiological literature. The current notions may best be set forth by a few quotations. Donaldson has written that "the capacity for undergoing expansive change is transient, and that those cells which fail to react during the proper growing period of an animal have lost their opportunity for ever." This has like- wise repeatedly been urged by Rubner. We do not know in truth, he says, whether Nature demands an absolutely uniform daily growth, or whether remissions in growth are permitted or even advantageous. This alone is certain, that interference with the growth impulse should not last during the entire period for growth ; otherwise the size of the individual will suffer permanent detriment. " Verlorene Körpergrösse in der Jugendzeit kann nach Vollendung der Wachstumsperiode nimmermehr abgeglichen werden." In his entertaining book on The Problem of Old Age, Growth and Death, Minot has hinted that some factors other than nutrition and age may enter into the question here discussed. "When the cell is in the young State, it can grow rapidly ; it can multiply f reely ; when it is in the old State it loses those capacities, and its growth and multiplication are correspondingly impeded, and if the Organi- zation is carried to an extreme, the growth and the multiplication of the cell cease altogether. We find, however, that there is some- thing a little more complicated yet to be considered, for it is not merely a question of the capacity of the cells, bat also of the exer- 164 Viewpoints in tJie Study of Growth [Jan. eise of that capacity, which we must deal with. Here comes in a factor which we learn from the study of regeneration." Experiments which have been conducted for some time by my colleague Dr. Thomas B. Osborne and myself, imder the auspices of the Carnegie Institution of Washington, make us hesitate to accept some of the older dicta respecting the hmitation of growth to a very definite period of Hfe. We have secured clear evidence that the growth of rats and mice may be suppressed or held in abey- ance for very long periods, even beyond the age at which any marked increment of size ordinarily occurs, without loss of the capacity of subsequent growth under appropriate conditions of diet. It is necessary to distinguish clearly between growth and regenera- tion or repair; for the latter is admittedly observed in the adult period of life. The natural cessation of growth is a fact familiär to everyone. As growth proceeds and the powers of the individual mature, his tendency to grow rapidly declines. This is an interesting phenome- non that has not been explained. Minot in particular has promul- gated the view that there is from a very early period a marked fail- ing in the capacity to grow, due to factors in the animal body itself . The notion that senescence finds its beginning in the very earliest periods of life is not a new one. Thus, Thomas Cogan, author of The Haven of Health, writing in 1596, says : "And if we do con- sider well the State of mankind in this life, we may see that a man beginneth to die as soone as he is borne into this world, for that the radicall moisture which is the roote of life, can never be restored and made up againe, so good as it was at our nativitie, but continr ually by litle and litle decaieth untill the last end of our life. Yet by that moysture which commeth of nourishment, through meate and drinke, it is preserved and prolonged, so that it is not so soone wasted and consumed as otherwise it would be. Like as a lampe by powring oyle moderately, the light is long kept burning, yet it goeth out at the last. And this is it which Hippocrates speaketh: The same heat which brought us forth consumeth us. Yet in the beginning of our age while nature is yet strong, more of the nour- ishment is converted into the substance of the bodie, than is con- sumed : and that while the bodie increaseth and groweth. After- ward so much only is restored as is wasted, and then the bodie is in 1914] Lafayette B. Mendel 165 perfect growth. At length nature waxing weaker, is not able to restore and repaire so much as is wasted and decayed, whereby the body beginneth to decrease, and the powers and strength thereof be more and more diminished imtil such time as hfe, even as the Hght of a lampe, be cleane extinguished. And this is called naturall death, which few attaine unto, but are prevented by death casuall, when by sicknesse or otherwise the said naturall moysture is over- whelmed and suffocate. Now the meanes to preserve this naturall moisture, & consequently to preserve life, is to use meates and drinkes according to the age of the person. For the dyet of youth is not convenient for old age, nor contrariwise." According to Minot, the more rapid growth depends upon the youth of the individual — its small distance in time f rom its procrea- tion. Cessation of growth is thus associated with age. We are told that there is " a certain impulse given at the time of impregna- tion which gradually fades out." The facts of regeneration in the lower forms warn us, however, to be cautious in drawing conclu- sions as to the loss of growth power. Something may inhibit it as age progresses. It has even been alleged that growth is stopped because an animal can digest only a limited quantity of food, and that the adult size is the stage of equilibrium between the amount of food digested and the amount used up. Experience in the field of nutrition speaks against such an assumption. If we turn our attention to the unit of biological activity, the cell, in seeking something more specific respecting the decline or cessation of growth, certain general principles may be drawn into consideration. In a unicellular organism under favorable condi- tions of nutrition the process of building up exceeds the destructive process so that the body increases in size up to a limit where fission takes place. " What determines the limit is unknown, but the cause is perhaps in some way connected with the geometric principle that the volume of the cell increases as the cube of its diameter, whereas the surface by which it absorbs nutriment and otherwise comes into relations with the outside world increases only as the Square of its diameter." Since activity is a function of the surface the larger the unit the smaller must be its activity. An organism can only attain a large size on this basis, by a multiplication of units, each present- ing the same amount of surface as an individual cellular organism. i66 Viewpoints in the Study of Growth [Jan. though it may be exposed to an internal rather than an external medium. One of the secrets of the cessation of growth in higher forms Hes in the hmitation of this cell division. A further aspect of the cessation of growth concerns the rela- tions of nucleus to cytoplasm in the growing cellular masses. The functions of nutrition and growth in cells depend upon the presence of a nucleus. Cells which attain a very large size may contain many nuclei. The process of cell division is determined by the ratio of the mass of nuclear chromatin material to that of the protoplasm of the cell. Jacques Loeb has pointed out that after fertilization there is an enormous synthesis of nuclear material. He suggests the pos- sibility that the ratio determining cell division may be determined by the laws of mass action and chemical equilibrium. This syn- thesis of nuclear Compounds from the protoplasmic constituents is a reversible process, according to Loeb. If there are continual readjustments to dispel the disproportion between nuclear and pro- toplasmic material, the cessation of growth may be correlated in some way with the final establishment of equilibrium according to definite chemical laws. Abnormalities of growth are attributable to manifold and di- verse causes. Constitutional defects, faulty nutrition, and various environmental factors may be more or less responsible. It is cor- respondingly difficult to classify on an etiological basis the mani- festations which they occasion. There occur deviations which are presumably still within the realm of the normal. This applies, for example, to the variations in the growth of the individuals of a litter, or between different litters. Anomalies of growth expressed by an exaggeratcd rate are among the rarities of physiology. In- stances of early gigantism which might fit this category have been described. The more frequent cases of rapid growth usually rep- resent recovery or response to suppressed growth rather than actual growth de novo. In other words, whenever the growth of an entire organism as well as that of individual organs is modified in the sense of acceleration this usually involves the reversal or return of a morbid condition to the normal. Inhibitory features of growth may manifest -themselves by ab- normally diminished growth, untimely complete cessation of growth (Wachstumsstillstand), or even decrease in size. Defects of nutri- 1914] Lafayette B. Mendel 167 tion are a common cause of slow growth; but even with adequate diet there may arise a condition of maintenance without growth. Here we enter the realm of dwarfed or stunted individuals. The complete suppression of growth has been accompHshed by Osborne and myself in a variety of ways which are not primarily attributable to underfeeding. The energy factor, as such, thus drops out of the problem. In this respect the experiments are not comparable with those of Waters and of Aron, both of whom accompHshed their results by underfeeding with adequate food materials. In our ex- periments the " energy requirement for maintenance " and the "energy requirement for growth," which together are essential to the developing organism, were both supphed. Our dwarfed rats did not grow primarily at the expense of stored tissue mate- rials : in respect to gross form they apparently f ailed to grow in any sense. If it is true that growth can only continue when the energy intake exceeds the mere maintenance requirement, it is equally true that an excess of calories does not per se insure growth in a suitable animal. This fact furnishes an opportunity to the in- vestigator to ascertain and difTerentiate some of the essential quali- tative factors : protein, inorganic salts, etc. — their minimum and Optimum values. Progressive decline in weight, negative growth (kataplasia), is an obviously pathological manifestation. Like most of the abnor- malities of the adolescent period its clinical manifestations are of decided importance. Irregularities of growth in the individual tend to be followed by compensating processes. Statistical studies on children, for in- stance, indicate that retardation in early growth is made up by abnormally rapid growth later. Whether all growth really stops in such instances, or whether our measurements do not merely indi- cate a loss of special body substances such as stored nutrients, needs to be determined. The answer has a bearing on the question already raised whether in the recovery process we are really dealing with new growth or with restitution of depleted tissues. Attempts to influence growth by drugs, such as alcohol, nicotine, caffeine, etc., are recorded in the literature. They need not be de- tailed in this connection. It should be remembered that the obvious efifects are not the only ones which may enter into the pathology of i68 Viewpoints in the Study of Grozvth [Jan. growth; defects may occur iinrevealed in the curves or gross mani- festations, yet involve the finer structures of the organism which are hidden to the naked eye. The use of the term growth with its several connotations has not infrequently led to a confusion with phenomena which are in reahty distinct therefrom. The distinctions are freqiiently subtle, yet none the less important. Thus the pathologist employs the terms hypertrophy and hyperplasia, the latter type of enlargement being the one dependent on the formation of new cells. In mam- mals, for example, there is no hyperplasic growth in the nervous elements after birth. Other tissiies, such as the connective and epithehal varieties, show abundant hyperplasia. The increase in size or weight that is observed in adult life has also been correlated with growth. The processes are presumably distinct. This addition to the organism or deposition of new mate- rial therein — the " Körperansatz " or " Mast " of the Germans — plays an important part in the "finishing" of cattle for the market. The "Ansatz" may be of a more permanent type such as charac- terizes the deposition of fat, glycogen or even protein in the cells; or it may be decidedly " unstable," representing water largely rather than food elements assimilated in proper proportions. This latter aspect of the storage depots in the body, in which water is present in exaggerated amounts, is not infrequently met with in pathology. True "Ansatz" must be distinguished, furthermore, from repair (reparation, realimentation, reconvalescence, recuperation, recov- ery) which takes place when by inanition, disease, or malnutrition, or all combined, growth has been checked in the adolescent or body weight lost in the adult. The increase in size attending realimen- tation has not infrequently been confused with true growth. They may be, and presumably are, distinct processes. In the one case depleted structures are restored, in part by mere deposition and restitution of the storage depots ; in growth, novel changes are ini- tiated in addition. There are indications available that the chem- ical and metabolic processes of repair are by no means identical with those of growth. Growth and regeneration undoubtedly have much in common; yet they deserve individual treatment. Quoting Morgan, "the word ' regeneration ' has come to mean, in general usage, not onl}'' 1914] Lafayette B. Mendel 169 the replacement of a lost part, but also the development of a new whole organism, or even a part of an organism, from a piece of an adult, or of an embryo, or of an Q.gg. We must include also those cases in which the part replaced is less than the part removed, or even different in kind. . . . The term ' physiological regeneration ' I shall use in the ordinary sense to include such changes as the moult- ing and replacement of feathers, etc. — changes that are closely re- lated to the life cycle of the individual." The power of regenera- tion in the general sense diminishes as we ascend the vertebrate Scale. Morgan believes that at least one reason for this lies in the lack of coördinate regeneration in the higher vertebrates, i. e., the slowness of certain tissues to regenerate in time with the other tissues. Inasmuch as this has an indirect bearing upon the possible causes of the uncorrelated growth which manifests itself as a path- ological phenomenon at times, it may be worth while to quote Morgan's view in some detail. He writes : " The evidence indi- cates, I think, with some probability, that the failure is due to the fact that the different tissues have very different rates of regenera- tion. In other words, each tissue in man seems to possess the power to regenerate its kind, but not all at the same pace, hence they fail to coöperate at the proper time to form a new structure. In man the skin regenerates; the muscles regenerate, though less well, per- haps; the nerves and the blood vessels regenerate, and the bones even have a not inconsiderable power to mend and even to some extent to regenerate. Hence, as I have said, the failure of the new limb to develop does not appear to be due to the failure of the indi- vidual Clements to regenerate, but is due to their failure to regen- erate concurrently. The bones or the ner^^es or the muscles may be the main cause of the trouble, for they produce new material with great slowness." The entire field of the physiology of repair is largely unexplored as yet. Losses due to simple inanition can apparently speedily be made good. Whether the end result as regards the composition of the restored tissues is the same as that pertaining in normal growth remains to be seen. If it is true that the capacity for regeneration implies a latent youthfulness the extent to which loss in weight is compatible with continuation of life may in part be dependent on the latent power of growth as well as repair. Practically this finds I/o Viewpoints in the Study of Growth [Jan, its expression in conclusions such as that the younger the child the more readily is rectiperation accompHshed. It appears to be a fact of experience that animals retarded in growth by underfeeding, as well as yoting children recovering from prolonged illness, begin to grow with more than normal vigor on a return to normal diet and health conditions and at the end of the growing period may be as heavy and have as heavy a brain as their normal companions. Nev- ertheless this will not yet justify us in concluding that the restored individuals are in all respects normal. Qualitative changes not ap- preciated by the cursory examinations may have become perma- nently engrafted. In all of the foregoing discussion of the phenomena of growth, its varied aspects, the criteria, the modifying factors and related phe- nomena, little has been said of the Initiation of this fundamental manifestation of living organisms. What is the underlying cause of growth? Like other biological processes this one can as yet be defined satisfactorily only in terms of its manifestations. The cause of regeneration is loss of body substance. It may frankly be admitted at the outset that we know almost nothing in regard to what takes place in protoplasm during growth and very little regard- ing the causes which incite or inhibit it. The only justification for veiling this ignorance in a vague terminology lies in the help which f ormulated hypothesis often brings to the Solution of obscure Prob- lems. With this prefatory Statement growth may be defined as the resultant of an inherent growth impulse — an internal factor — and a suitable environment, the latter including the food supply — an external factor. In these are concerned certain typical biological forces, the metabolism of matter and energy, as well as certain physio-chemical reactions. The conditions determining growth are mainly resident in the cells. If we are unable today to define the internal factor suitably, it is equally impossible to cite all the external factors as we do for plants. Air, light, warmth, food, etc., have their functions. How- ever essential food may be for growth — and no one can gainsay its preeminent importance — it can in no sense be regarded as the su- preme cause of growth. Nutrition can only give the growth im- pulse free play. This factor can, however, be subjected to experi- mental analysis. The role of the individual nutrients, — organic and 1914I Lafayette B. Mendel 171 inorganic, — the energy features and other nutritlonal details can be studied with some precision. But of what we have called the internal factor in growth — the growth impulse, the tendency to grow, the capacity to grow, " Wachstumstrieb," " Wachstums fähig- keit," " WachstumsmögHchkeit," — the factor which is hereditary in its origin and sets to growth the hmits which nutrition cannot f un- damentally alter, httle further can be said. It may be that the rythm of cell division and its attendant features, which some have identified as the detectable expression of the capacity to grow, is dependent upon the action of " hormones," products of internal secretion and cellular metabolism. If this is true we shall have at our disposal some means of modifying the internal factors of growth. For the present we must content ourselves with the un- satisfying conclusion that to unravel the inner nature of growth is to penetrate the secret of the distinguishing characteristics of living substance. Here hypotheses reign uncontrolled. However dominant the röle of the cell in growth may be, the problem of development cannot be investigated solely from the Standpoint of cellular physiology. We may admit the limitations of nutrition in furnishing an adequate explanation of either the " Wachstumstrieb " or the " Wachstumsmöglichkeiten " ; but in any event the food factor in growth is one that is open to experimental study. The greatest hope of advance in the Solution of the obscure questions of growth therefore appears to lie in this direction. Along this direction, for example, Osborne and I have found with our coworkers that not all proteins suffice to promote growth. Some of them are apparently adequate to fulfill the needs of both maintenance and growth; others like gliadin satisfy the requirement of maintenance alone ; while such " incomplete " proteins as zein and gelatin are by themselves inadequate in every sense for perfect nutri- tion. The incapacity of some of these proteins unquestionably lies in a lack of certain essential amino-acid units. It must be noted that growth has not been accomplished with any protein lacking the cyclic groups such as are found in tyrosine, Phenylalanine, and tryptophane; and we have lately found that lysine is indispensable for tissue construction. That these " inefficient " proteins are not primarily toxic to the organism is shown by the fact that we have found many of them to be adequate for maintenance in both grow- 1/2 Viewpoints in the Study of Growth [Jan. ing and adult organisms; whereas, as is well known, others like zein and gelatin by theniselves do not even suffice for this function. The need of an adeqiiate siipply of energy in some form or other, and of appropriate salts for tissue construction is obvious. The importance of the latter gains an unsuspected proniinence when one plans experiments with isolated food substances; for with the ordinary natural food mixtures a reasonable modicum is already provided. We are convinced from an extensive experience that many failures to promote growth in experiments on artificial nutri- tion have centered in the inorganic food ingredient of the selected dietaries. In some instances the deficiencies, if such there are, may involve some minute proportion of hitherto supposedly inessential Clements like iodine, manganese, etc. The time and opportunity for investigation along the lines here suggested has arrived. There is, further, a considerable body of evidence to suggest that "hor- mones " or " vitamines " or comparable stimulants of growth are essential. It is f utile as yet to discuss their mode of action. In any event, however, their röle among the external causes of growth need not be that of a simple nutrient in the sense of yielding energy or material for development. The analysis of growth into a controllable nutrition factor and an inherent growth impulse has its significance for the appreciation of the pathology of growth and the management of the situations created thereby. It becomes clear that abnormal growth cannot always be corrected by regulation of the external factors of diet, etc. A limit to dietotherapy or other therapeutic measures is ofttimes set by the inalterable inheritance of an imperfect Constitu- tion, the basis of an adequate capacity to grow. To attempt to f ormulate " laws " from data which are not over- abundant and which involve a considerable number of variables can scarcely be expected to yield generalizations of a very exact nature. Nevertheless there have been essays at probable laws for growth. As an Illustration I may cite Lusk's generalization that " during the normal development of the young of the same age and species, a definite percentage of the food is retained for growth irrespective of the size of the individual." Rubner has independ- ently applied a quite similar law to the growth of all species except man, as an expression of the belief that it requires the same energy 1914] Lafayette B. Mendel i73 equivalent to construct a unit of new substance in young animals. Rubner has also formulated conclusions regarding the length of life of individual species in relation to the number of calories metab- olized during life. Exact mathematical expressions have already been devised for certain features of growth. In the light of our scattered knowledge I can only agree with Friedenthal that in biology it is best to avoid the term "law" and content ourselves with generalizations which have their exceptions and are not abso- lute in their comprehension. We are still far from the stage where laws of growth can successfully be propounded. The path which the natural sciences have successfully pursued in the past has not led from proposed laws to facts, but rather in the reverse direction from the collection of facts to generalized rules. This is likewise the way which the research of the future must follow. It is almost impossible to review the salient features of the phys- iology of growth without directing attention to the obvious gaps in our knowledge thereof. Problems await us at every turn, — some of them clearly defined and open to experimental investigation, others obscured in the haze of conflicting data or complicated by the manifold factors which enter into the questions of development. No review of any aspects of a progressive and growing science is complete or illuminating unless it suggests problems as well as answers them. A passing reference to some of them may not be inappropriate here. Statistics of growth being the easiest to ob- tain, have been collected in greatest number. But there are numer- ous Statistical details involving the growth of the individual parts or Organs which are quite as essential for the understanding of what is involved in the correlation between organs incidental to growth. More than ever the need of additional studies of the histology of growing tissues in mammals looms up at the present time. They are fundamental to any investigation of the morphological back- ground against which the growth functions are projected. The popularity which histological investigation has enjoyed in embry- ology in recent years should be extended into the postnatal stages of life. In general we are taught that different tissues have unlike power of growth in the sense of cell multiplication. Some, like the testes, multiply their cells throughout life; others, like the muscles 1/4 Viewpoints in the Study of Growth [Jan. and nervous System, only in the embryonic period; others again, like the glands, diiring a variable period after birth. So long as so much emphasis is placed upon the cell in growth we deserve to know more about its morphological history in this crucial period ; and this applies above all to those tissues, the so-called ductless glands, like the hypophysis and thymus which it is currently customary to asso- ciate in some way with growth. Until very recently the interest of histologists has centered either in the embryonic or the adult tissues of mammals, with scanty consideration of the intervening stages. The importance of a comprehensive consideration of nutrition in growth need not be dwelt upon in detail. The conflicting views which have been held since Liebig's time regarding the significance of protein in nutrition have had their counterpart in the explanation of growth. Voit assumed that the protein metabolism of the grow- ing organism is unlike that of the adult. The inadequacy of the theories which associate growth with a decreased power of protein katabolism no longer requires emphasis. Now that we know of the marked chemical and biological differences existing between pro- teins from different sources the significance of these facts in nutri- tion must be further established. Important economic considerations are involved in the ability to modify growth or accomplish it at lessened expense, — a possi- bility for which nutritive factors offer the only probable oppor- tunity at present. Hence arises the practical importance of some of the Problems in this field. Broadly put, one problem reads : How can inefficient native f oods be made efficient, and what is the relative economy of different dietaries and adjuvants ? The questions which arise in connection with the duration of the capacity to grow have already been alluded to. Will growth, suppressed by the necessary restrictions in diet, cease entirely for an indefinite length of time ? What happens to an animal suffering such a Suspension of growth, when it is given an abundant diet? What are the tissue changes accompanying these suppressions and realimentations, i. e., what are the attendant histological features? The function of age is evidently an important matter for con- sideration in questions of growth. An index of age apart from the record of birth is much to be desired. Weight and size in gen- eral may be inadequate for many obvious reasons. What can be 1914] Lafayette B. Mendel 175 accomplished in this direction is indicated by the investigations of Donaldson and Hatai on rats. They have found that the percent- age of water in the central nervous System is a function of age and under ordinary conditions, whether the animal be over size or under size, well nourished or ill nourished, with a large brain or a small brain, the percentage of water remains practically unmodified by these conditions. The deviations in extreme malnutrition even are, very slight at most and completely disappear on a return of normal nutritive conditions. The percentage of water in the central nerv- ous System accordingly is the best index yet available of the normal process of senescence. The facts fit in with human experience in showing that hardships which include under feeding need not neces- sarily shorten the span of life. It is no small advantage to have a dependable index of age. The experimental possibilities and de- siderata in this field are far from exhausted. Chemical data are needed to Supplement the histological and other analyses of age. Although each individual appears to strive to attain a definite size it is still debated whether nature demands continuous growth or to what extent remissions are detrimental or permissible. It is becoming more and more evident that growth is not an entirely indispensable function of living matter. Otherwise stated, the failure to grow is not incompatible with life. Where are the limi- tations of such situations? And above all, can life be extended by the delay of growth? This is one aspect of the broad problem of prolonging life artificially by altering the conditions under which it goes on. Theoretically it is conceivable that growth processes, like chemical reactions, should be reversible. The more detailed study of growth may be expected in the future to bring helpful experience to bear on the manifestation of pathological neoplasms which concern health so vitally. The com- petition of youthful cells such as those of malignant tumors with the normal cells of adult tissues frequently brings victory to the younger tissue. The problems of normal development and ab- normal structure are doubtless in many respects one and the same. " Every great advance of science opens our eyes to facts which we had failed before to observe, and makes new demands on our 176 Viewpoints in the Study of Growth [Jan. powers of interpretation. . . . Great as the advance of scientific knowledge has been, it has not been greater than the growth of the material to be dealt with. The goal of science is clear — it is noth- ing Short of the complete interpretation of the universe. But the goal is an ideal one — it marks the direction in which we move and strive, but never a stage we shall actually reach. The universe grows ever larger as we learn to understand more of our own corner of it." Sheffield Scientific School, Yale University, New Haven, Conn. THE PHYSICO-CHEMICAL BASIS OF STRIATED MUSCLE CONTRACTION 3. The maximum surface tension in striated muscle WILLIAM N. BERG Introduction About a year ago, the writer^ publisht some calculations on the lifting power of striated muscle, in which an attempt was made to ascertain whether the changes in the surface tension between the contractu units and their surrounding medium were great enough to account for the hfting power of striated muscle. Among others, the assumption was then made that at the mo- ment when a striated muscle begins to contract against an external resistance, the surface tension between the contractu units and their surrounding medium (presumably the sarkoplasm) might possibly be as high as 85 dynes per cm. This was regarded as the upper limit, altho certain data in the literature, to be presented later, plainly indicated that the Upper limit could not be so high. Several assumptions were made in favor of the surface-tension theory for the purpose of justifying the temporary or provisional use of 85 dynes per cm. as the maximum surface tension between the con- tractu units and sarkoplasm. A minimal surface tension was like- wise assumed from the literature for the relaxation phase, but as this figure was purposely and provisionally omitted from the final calculations, it may be disregarded for the present. The results indicated that in a working striated muscle, surface energy can furnish but a small part of the total energy transformed, a conclusion diametrically opposed to that of Bernstein, Macallum^ and others, who regard a working muscle as a mechanism that can transform a quantity of surface energy equivalent to the external work done. 1 Berg: Biochemical Bulletin, 1912, ii, p. loi. 2 Macallum : Jour. Biol. Chem., 1913, xiv, p. 96. 177 178 Striated Muscle Contraction [Jan. As ftirther evidence of the correctness of the surface-tension theory, Macallum^ recently quoted Jensen'* to the effect that "a thread measiiring one mm. in diameter formed of the Plasmodium of Chondrioderma, a Myxomycete, may, when it is in the dense condition, bear up a weight of nearly a gram. If the force engaged is surface tension it would amount to about 6000 dynes per cm." Because of its enormity this figure of Jensen's at once arrests the attention, for nowhere in the annals of physics and chemistry, can a surface tension of 6000 dynes per cm. be found. Most organic substances that are fluid at ordinary temperatures, have surface tensions well below 50 dynes, ^ when measured against air. Very few aqueous Solutions have surface tensions higher than 85 dynes per cm. For a large number of molten salts the figures vary be- tween 100 and 200 dynes. For most of the metals, when melted, the figure varies between 250 and 1000, altho platinum seems to be exceptionally high, having a surface tension in the molten state of about 2000 dynes per cm.^ Presumably, the plasmodium of Chondriodeniia has a chemical composition similar to that of other living matter and consists essen- tially of water plus the usual materials present in living matter. The surface tension of water, when measured against air at ordi- nary room temperatures, is about 70 dynes per cm.''' Altho small amounts of some organic substances can lower the surface tension of water considerably, the surface tension of water cannot be raised very much by any substance, even when present in large amount. There seems to be an upper limit to the surface tension of aqueous Solutions (in exceptional cases this is as high as 109 dynes per cm. ^Macallum: Jour. Biol. Chem., p. iio. * Jensen : Anatomische Hefte, 1905, xxvii, p. 842. ^ If not otherwise indicated, figures for surface tension are in dynes per r dvnes centimeter, t. e.,-^ . The magnitude of the dyne can be easily grasped if it be cm. borne in mind that a mass of i gram is attracted to the earth with a force of i gram, or 981 dynes. Consequently, a dyne is practically equal to a milligram (force). ^FreundHch: Kapillarchemie, pp. 30, 62 (Leipzig, 1909). Landolt-Börnstein : Physikalisch-chemische Tabellen, 4te Aufl., pp. 1 14-129 (BerHn, 1912). ''Landolt-Börnstein: Loc. cit., pp. 112, 128, 129; Freundlich: Loc. cit., p. 62; Lewis: Ztschr. f. physikal. Chem., 1910, Ixxiv, p. 619; Heydweiler: Ann. Physik., 1910, xxxiii, pp. 145-185. 1914] William N. Berg 179 for very concentrated Solutions of potassium carbonate), which varies from 71 to 85 dynes per cm. for the aqueous Solutions of most inorganic salts. From the data in the literature, one might infer that the surface tension of Chondrioderma ought not exceed that of other aqueous Solutions or suspensions of comparable com- position. It should, therefore, be somewhere in the neighborhood of 85 dynes per cm., or below. But according to Jensen's calcula- tions, it is 6000 dynes per cm.. One reason for doubting the cor- rectness of this figure has already been mentioned, namely, the fact that surface tensions as high as this are not to be found recorded in the literature, for any substance. Secondly, Jensen apparently over- looked the fact that the circumference of a circle is 27r times the radius and not tt times the radius, which is the expression used in Jensen's^ formula, with the result that a quotient of 6000 ought to be 3000. A somewhat similar error was made in our own calculations already publisht,^ insofar as the value of tt was accidentally omitted from the value for the reduction in area when i cc. of muscle con- tracts. The value previously obtained, 1000 sq. cm., should be a little over 3000 sq. cm., with corresponding changes in the results. So many assumptions had been made in favor of the surface tension theory, that no change was necessary in the final conclusion, namely, that the surface-tension theory of striated muscle contraction as advanced by Bernstein, by Macallum and others, is untenable. It is rather remarkable that altho Bernstein's^^ calculations showed that surface energy was insufficient (p. 295), and he realized the insufficiency, he still advocated the theory as being correct in prin- ciple (p. 141). 8 Jensen, loc. cit., p. 841 : " Diese (the surface tension) ist berechnet aus dem Gewicht, das ein Pseudopodienbündel von bekanntem Gesamtumfang zu heben vermag. Die Berechnung geschah nach der Formel a = p/'^r, wo a die Ober- flächenspannung, p die Zugfestigkeit eines Pseudopodiums und r sein Radius ist." Jensen used this same formula in the two widely differing cases of a thread (Orbitolites) that lifts a weight and a thread (Chondrioderma) that sus- tains a weight. This is the probable reason for the great difference between the surface tensions of Orbitolites and Chondrioderma and not as proposed by Macallum {loc. cit., p. iii) : "It is not improbable, therefore, that surface tension may be very high in some forms of living matter and very low in others . . ." ^Berg: Biochemical Bulletin, 1912, ii, pp. 107-109. 10 Bernstein : Arch. f. d. gesammte Physiologie, 1901, Ixxxv, pp. 271-312; 1909, cxxviii, pp. 136-141. i8o Striatcd Muscle Contraction [Jan. Jensen's arithnieticall}^ correct figure of 3000 dyiies per cm. is still so very much higher than that of any known substance that one is led to suppose that perhaps Jensen's method of calculating the surface tension is incorrect. According to his formula, the weight sustained by a plasmodium thread when divided by the circumfer- ence of the thread gives the surface tension. This method may be correct, but it is not mentioned among the various methods described in the literature. This makes it desirable that someone prove its correctness. To the writer it seems that the quotient obtained by Jensen does not represent a surface tension. According to Pfeffer/^ the question of surface tension does not enter the problem (of Chondrioderma) at all, for the reason that the outer layers of the plasmodium thread of Chondrioderma are solid at the time when a weight can be sustained. Or, to be more precise, Pfeffer states that Chondrioderma have the property of reversibly varying the consistence of the outer layer, from that of the fluid protoplasm in the interior of the cell to that of solid gelatinous masses. The tougher outer layer is regarded by Pfeffer as a physiological product, and not until this has been brought back to its originally fluid condition can changes in surface tension be regarded as factors in the problem. If Jensen's formula gives results that have a real physical mean- ing, it ought to be possible to apply it to other forms of living mat- ter and to obtain results that can be interpreted. Parnas^^ found that the smooth muscles of certain clams could sustain very great weights. He so loaded living, intact clams, that in one case (p. 458) a smooth muscle having a cross section of 0.3 sq. cm. sustained a weight of 3000 grams for three hours. Similar observations on the great weight-sustaining power of smooth muscle have been made by others.^^ Assuming that the cross section was of uniform tensile resist- ance, each sq. mm. sustained a weight of 100 grams, which is 75 iiPfeflfer: Pflanzenphysiologie, 2te Aufl. (1904), pp. 716-718. 12 Parnas : Arch. f. d. gesammte PhysioL, 1910, cxxxiv, pp. 441-495. i^Bethe: Arch. f. d. gesammte PhysioL, 1911, cxlii, pp. 291-336. Cohnheim and von Uexkull : Ztschr. f. physiol. Chem., 1912, Ixxvi, pp. 314-321; Cohnheim: ibid., pp. 298-313. I9I4] William N. Berg i8i times the weight sustained by the i mm. thick plasmodium of Chon- drioderma according to Jensen's Observation, and which is quoted by Macallum as an example of high surface tension in hving matter. Since a thread of this smooth muscle i mm. in diameter can sustain 75 grams, the surface tension between it and its surrounding medium, water in this case, must be over 468,000 dynes per cm. according to Jensen's formula. This value becomes 234,000 dynes per cm. if Jensen's formula be used as it probably was intended to be used (see p. 179). The correctness of such a figure and of the method used in obtaining it are to be doubted because the surface tension calculated in this way is greater than that of any other known sub- stance. The problem of the transformation of energy in striated muscle is, in part at least, a problem in dynamic mechanics. It seems Strange that certain investigators^^ should attempt to treat the subject as if it were a problem in static mechanics. The Plas- modium of Chondrioderma did not lift a gram, it did no work, for the weight was only sustained, and in this respect the phenomenon is comparable with the sustaining of very much greater weights by smooth muscle, but is not comparable with the lifting of weights by striated muscle. That the plasmodium could sustain a weight be- cause of the surface tension between its surface and the surround- ing medium is for the present purely an assumption, for neither Jensen nor Macallum have brought f orward any evidence showing that surface tension was a factor in the problem, THE SURFACE TENSION BETWEEN CONTRACTIL UNIT AND SURROUNDING MEDIUM To know the limiting values of the surface tension between the contractu units and the surrounding medium is obviously of the greatest importance in connection with the surface-tension theory. Fortunately, the data in the literature on the surface tension be- tween two liquids or between two Solutions are sufficiently complete to point definitely to the limiting values desired. Two Solutions are to be considered: (i) The Solution of biological substances which constitutes the lateral surface of the contractu units and (2) 1* Bernstein: Arch.f.d.gesammte FÄy^yio/., 1901, Ixxxv, pp. 271-312. Jensen: Anatomische Hefte, 1905, xxvii, p. 842. i82 Striated Muscle Contraction [Jan. the Solution of biological substances which bathes these contractu Units, and which probably is tissue lymph. It is assumed of course, that the transformation of energy into external work takes place on the lateral surfaces of the contractil units and not on their upper and lower bases (otherwise a muscle would be stronger crosswise than it is lengthwise). These two Solutions are assumed to be in contact with one an- other thruout the contraction and relaxation phases and, presum- ably, the surface tension between these two media is due to the rapid chemical changes taking place hoth inside and oiitside the con- tractu unit}^ Insofar as the surface tension varies, being high just as contraction begins and being low ( ?) as relaxation begins, the chemical composition of the two Solutions must vary continually, even when the muscle is apparently at rest, for then the condition of tonus still requires the expenditure of energy, tho in smaller amount. How high can the surface tension be between contractil unit and lymph? It obviously cannot be higher than that of a saturated aqueous Solution of the salts found in living tissue. In general, the organic constituents of lymph and of blood serum tend to lower the surface tension of water, the inorganic constituents tend to raise it. The effects of the former predominate in blood serum. Morgan and Woodward,^® using very accurate methods, determined that the surface tensions of the blood sera of several kinds of animals, in- cluding man, were practically the same and that, at 37° C, it varied between 44 and 48 dynes per cm., when measured against air. That is to say, under ordinary conditions the surface tension of blood serum is two thirds that of water, both surface tensions being measured against air. Insofar as lymph and blood serum do not differ much in their composition, the above figure gives at least an idea of the value about which the surface tension in working muscle fluctuates. But the Solution on the contractil unit may differ in its composition, for very short periods of time, from that of the sur- rounding lymph and it may have a higher surface tension. ^^ Berg : Arch. f. d. gesammte Physiol., 1912, cxlix, p. 205. "Morgan and Wood ward: Jour. Amer. Chem. Soc, 1913, xxxv, pp. 1249- 1262. For further data on the surface tensions of serum, lymph, etc., see Neu- berg : Der Harn sowie die übrigen Ausscheidungen und Körperflüssigkeiten, ii, p. 1724 (Berlin, 1911). I9I4] William N. Berg 183 The surface tension may be raised by at least two processes : (i) Substances that lower the surface tension of water are re- moved, thereby bringing the surface tension up to that of water, and (2) inorganic salts are at the same time brought into the Solu- tion, thereby raising it beyond that of pure water. For the present it may be assumed that, at the beginningof the contraction, the con- tractu Unit is covered with a layer of saturated sodium chlorid Solu- tion, because sodium chlorid is the most abundant inorganic salt present, and its saturated aqueous Solution has as high a surface tension against air, i. e., about 85 dynes per cm., as that of any other Solution of biological substances. Concentrated Solutions of sodium hydroxid and of potassium carbonate^ ''^ are exceptional insofar as their surface tensions are somewhat higher than that of saturated sodium chlorid Solution. These may be disregarded because of their absence f rom living matter. It is altogether possible, and in fact quite probable, that the changes just described do not take place on the contractil unit. It should be borne in mind that the object of considering a saturated sodium chlorid Solution as covering the contractil unit just before contraction is solely for the purpose of justifying the provisional use of the highest surface tension recorded in the literature for the aqueous Solutions of biological substances. Eighty-five dynes per cm. is not a postulated value for the surface tension of the Solution on the contractil unit at any time. What the actual highest value really is, may, for the present, be regarded as an unknown quantity. Eighty-five dynes per cm. is an Upper limit for all the Solutions in the active muscle — in which Solutions the chemical and physical changes take place that presumably underlie the vital activities of striated muscle.^ ^ Eighty-five dynes per cm. is the surface tension of saturated sodium chlorid Solution against air at 18° C. What is the surface tension of saturated sodium chlorid Solution against lymph? Pre- sumably, the surface of the contractil unit is at all times in contact with another liquid, the lymph, and not with a gas. In the calcula- i'' Landolt-Börnstein : Physikalisch-chemische Tabellen, i\it A.Vi^.,p. 12g (Ber- lin, 1912). 18 Freundlich: Kapillarchcmie, p. 62 (Leipzig, 1909). Landolt-Börnstein : Physikalisch-chemische Tabellen, 4te Aufl., p. 129 (Berlin, 1912). 184 Striatcd Muscle Contraction [Jan. tions previously publisht^^ the assumption was made, in order to give the surface-tension theory the widest latitude, that the surface tension of the contractu sokition {i. e., the Solution on the lateral surfaces of the contractil iinit) against lymph is the same as it is against air, or in other words that the surface tension of lymph against air is zero. But this, of course, is not true. Lymph has a surface tension not very dififerent from that of serum. The theoretical maximum effective surface tension between the contractil Solution and the immediately adjacent layer of lymph can be ascertained from ( i ) the maximum surface tension of the con- tractil Solution against air; (2) the minimum surface tension of the adjacent lymph against air, and (3) the assumption that these two occur simultaneously at the beginning of the contraction phase. The second value subtracted from the first gives the figure desired. The surface tension between two liquids is equal to, or is less than, the difference between their surface tensions when measured sepa- rately against air,^*^ provided they do not react chemically. For reasons already mentioned, the maximum surface tension of the contractil Solution, against air, has been assumed to be 85 dynes per cm., or a value very close to it. In a similar way, the lower limit for the surface tension of the adjacent lymph, when measured against air, may be assumed to be the lowest surface tension re- corded in the literature for aqueous Solutions of biological sub- stances.^^ A dilute Solution of sodium oleate, or a concentrated Solution of a fatty (butyric) acid,^^ both have very low surface ten- sions, very near 26 dynes per cm. Either of these Solutions may be assumed to constitute the adjacent lymph at the moment when the contractil Solution has its maximum surface tension. The surface tension between the contractil Solution and the adjacent lymph, assuming that there is such a surface tension, would be the differ- 1^ Berg : Biochemical Bulletin, 1912, ii, p. 107. 20 Quincke: Annalen der Physik und Chemie (PoggendorfF's Annalen), 1870, cxxxix, pp. 1-89; Whatmough; Ztschr. f. physikal. Chem., 1901, xxxix, p. 175; Antonow : Jour. de chimie physique, 1907, v, p. 372. 21 An alternate assumption is possible: that the adjacent lymph is momen- tarily separated from the contractil Solution by a layer of pure organic substance having a very low surface tension, i. e., butyric acid, 26, or acetic acid, 23, dynes per cm. 22 Freundlich : Kapillarchemie, pp. 56, 59 (Leipzig, 1909). 1914] William N. Berg 185 ence between the surface tensions of these Solutions measured sep- arately against air, i. e., 85 minus 26 or very nearly a maximum of 60 dynes per cm. What is the surface tension between contractu Solution and adjacent lymph at the beginning of the relaxation phase ? The most expedient answer is, zero; a condition which can arize when con- tractu Solution and adjacent lymph have acquired the same compo- sition, altho in certain cases (see p. 190) the surface tension may be zero between two Solutions differing in their composition. The muscle relaxes presumably because its own weight is sufficient to re- form the contractu surfaces against zero surface tension. This as- sumption taxes the theory least. If the surface tension during the relaxation phase is anything other than zero a second problem arizes which is as great as the first, namely, to explain how a relaxing muscle, presumably doing no work, can, in the act of relaxation, re-form the large contractil area against a surface tension greater than zero. Naturally, the greater the surface tension at the begin- ning of the relaxation phase, the more work must be done in re- forming the contractil area against the surface tension. To summarize : In order that there may be a surface tension between the contractil Solution and the adjacent lymph, it is neces- sary to assume a difference in chemical composition between the two. The differences assumed are such as to give the greatest sur- face tension — about 60 dynes per cm. The composition of the two Solutions cannot be varied sufficiently to give a surface tension any where near 6000 dynes per cm., which is the surface tension be- tween certain forms of living matter and water, according to Jensen and Macallum. The conditions in a working striated muscle, as just described, can be theoretically pictured as f ollows : Immediately before the contraction phase begins the lateral surfaces of the contractil units or muscle rods are covered with saturated sodium chlorid Solution. Immediately in contact with this is the adjacent lymph consisting of a concentrated Solution of butyric acid. Under these conditions the contraction begins with a surface tension of 60 dynes per cm., pro- vided, as already pointed out, these maximum and minimum values occur simultaneously and at the beginning of the contraction phase. If the contraction began under the driving force of 85 dynes per i86 Striated Muscle Contraction [Jan. cm., and if this were maintained thruout the entire contraction phase, and if striated muscle were a mechanism having an efficiency of loo per Cent., then the surface energy in i cc. of muscle would enable it to lift a little over looo grams^^ and surface energy might be regarded as the cause of muscle contraction. The efficiency of the muscles of the human body as a machine is very close to 21 per cent.,^^ and it is almost certain that if any of the driving force in striated muscle is due to surface tension at all, the tension is but a small fraction of 85 dynes. The above figures should be interpreted accordingly. Washington, D. C. 23 Berg : Biochemical Bulletin, 1912, ii, p. 109. The value, 361 grams, is to be multiplied by 3.1416, the value of tt having been accidentally omitted. The correct figure is, therefore, 1134 grams. 24 Benedict and Carpenter : Bulletin 208, Office of Experiment Stations, U. S. Dept. of Agric, 1909, pp. 1-44. Hill : Joiir. of PhysioL, 1913, xlvi, pp. 435-469. THE PHYSICO-CHEMICAL BASIS OF STRIATED- MUSCLE CONTRACTION 4. Sources of surface tension in striated muscle WILLIAM N. BERG Thruout the works of Bernstein, Jensen and Macallum on muscle contraction, the assumption is made (or implied) that the chemical changes taking place in an active muscle are such as to give rise to a surface tension between the contractu unit and the adjacent lymph, as if a difference in chemical composition or in concentration between two adjacent regions necessarily causes sur- face tension. Which chemical reactions in muscle can cause sur- face tension between contractil unit and adjacent lymph? That there are very f ew such reactions will, perhaps, be apparent f rom the following data on the surface tensions of aqueous Solutions of bio- logical substances. Suppose two adjacent regions in a living cell to differ in their chemical composition. Will there be an osmotic difference between the two? The answer is that this will depend upon the nature of the difference in composition. If the two regions differ only in the fact that there is more protein present in one than in the other, there will be no osmotic difference, or the difference will be a negligible one. But if the regions differ in their concentrations of electro- lytes, or of organic substances of relatively low molecular weight, there will be an osmotic difference which may be very great. That is to say, some solutes exert osmotic pressure, others do not. The same is true of solutes with regard to surface tension. Whether a difference in chemical composition or concentration between two adjacent regions necessarily involves surface tension depends upon the nature of the solute. For the present purpose, mbst Solutions of biological substances may be considered as falling into one of the following three classes. (Certain data on the surface tension of these types of Solutions will 187 i88 Striated Muscle Contraction [Jan. be considered with reference to their bearing on the problem of the sources of surface tension in living matter in general and in striated muscle in particular. ) The first class of Solutions to be considered are those which are obtained when two partly miscible liquids, such as ether and water, for example, are thoroly shaken, and then allowed to separate into two layers, forming in this case, a Solution of water in ether and a Solution of ether in water. The surface tension between two such layers is of interest for several reasons, principally because such binary mixtures probably exist in living tissue, which is supposed to consist of a comparatively dilute aque- ous Solution or Suspension of the biological constituents that bathes the more viscous protoplasmic structures. These structures, what- ever their shape and size may be, are commonly regarded as Solu- tions of water in the biological constituents. The surface tension between two liquids was studied by Quincke in 1870.^ He found that the surface tension between two liquids, immediately after they have come in contact, is very nearly equal to the difference between their surface tensions when measured sepa- rately against air. For example, the surface tension of Chloroform against air is 30.6 dynes per cm., that of water against air is 80.9; the surface tension between Chloroform and water is approximately the difference between these two figures, or 29.5 dynes per cm. If the two liquids were mutually soluble, the surface tension between the two decreast to an extent which depended upon their mutual solubility and other factors. The following typical data, from Quincke's paper, are of interest because the general principle de- duced from them by Quincke is directly applicable to the conditions existing in a working muscle (on the assumption that the chemical and physical changes underlying muscle contraction take place be- tween Solutions or other liquids and not between Solutions and gases). Quincke, loc. cit., p. 27. Table X. Results at 20° C. in dynes per cm. In the first column of figures is given the surface tension of the first liquid against air ; in the second column, the surface tension of the 1 Quincke: Annalen der Physik und Chemie, 1870, cxxxix, pp. 1-89. (Pog- gendorf's Annalen.) 1914] William N. Berg 189 second liquid against air; in the third column, the surface tension be- tween the two liquids as actually determined.^ Carbon disulfid-water 32.0 80.9 41.7 Petroleum-water 31.7 80.9 37.6 Chloroform-water 30.6 80.9 29.5 Olive oil-water 36.8 80.9 20.5 Turpentine-water 29.7 80.9 11.5 Olive oil-alcohol 36.8 25.5 2.2 Quincke concluded that the surface tension between tvv^o liquids that do not react chemically, of course, is not quite equal to the difference betv^een their surface tensions when measured separately against air, and that it varies between this difference and zero, ac- cording to the mutual solubility, etc., of the two liquids (p. 18). This principle is important, because the surface tensions between Solutions probably existing in muscle can be easily calculated, if the surface tensions of these Solutions have been measured against air. A direct measurement of the surface tension between contractu Unit and adjacent lymph is, therefore, not absolutely necessary. Antonow^ measured the surface tension between water and a second liquid, such as Chloroform, ether, etc., practically repeating some of Ouincke's work, with the same result, i. e., the surface tension between two liquids is equal to the difference between their surface tensions when measured separately against air (p. 384). In certain cases, studied by Whatmough^ and by Antonow,^ the mutual solubility of the two liquids may be great enough to bring the surface tension down to zero or very near to it, while the two liquids still differ greatly in their composition. Whatmough (p. 178) studied the surface tensions of several binary mixtures, such as phenol and water, isobutyric acid and water, etc. In his experi- ments, the two liquids were thoroly mixt, allowed to separate into layers, and then portions of the upper and of the lower layers were separately removed and their surface tensions against air deter- - An extremely interesting and detailed summary of data on surface ten- sions of Solutions, etc., was also given by Castell-Evans : Physico-chemical Tables, ii, pp. 708-801 (London, 1911). ^Antonow: Journal de chimie physique, 1907, v, pp. 362-385. * Whatmough : Ztschr. f. physikal. Chem., 1901, xxxix, pp. 129-193. 5 Antonow : Loc. cit. 25 °C. 40 °C. 20 °c. 40 ''C. 35 "C. 35 °c. 190 Striated Muscle Contraction [Jan. mined. The following results are typical of many others obtained by Whatmough (p. 181).® Phenol and Water 100 gm. water layer contain 8.5 gm. phenol 42.5 dynes/cm. 100 gm. water layer contain 9.6 gm. phenol 41. i dynes/cm. 100 gm. phenol layer contain 72.1 gm. phenol 42.4 dynes/cm. 100 gm. phenol layer contain 66.9 gm. phenol 40.6 dynes/cm. Anilin and Water 100 gm. water layer contain 3.4 gm. anilin 57.0 dynes/cm. IOC gm. aniHn layer contain 95.1 gm. anilin 51.6 dynes/cm. Isobutyric Acid and Water 6.5° C. IOC gm. water layer contain 16.4 gm. isobutyric acid 29.9 dynes/cm. 25.2° C. 100 gm. water layer contain 36.3 gm. isobutyric acid 28.4 dynes/cm. 6.0° C. IOC gm. acid layer contain 73.4 gm. isobutyric acid 29.6 dynes/cm. 25.2° C. 100 gm. acid layer contain 36.3 gm. isobutyric acid 28.4 dynes/cm. From the above data it is evident that two aqueous Solutions in contact with one another may differ greatly in their concentration of a common solute, with a very small surface tension between them. In the above cases, the surface tension between the phenol and water Solutions is practically zero and the same is true of the isobutyric acid Solutions. Between the water Solution of anilin and the anilin Solution of water there is a surface tension of but 5.4 dynes per cm., altho the two layers differ in the fact that one contains 3.4 per cent. of anilin ; the other, 95 per cent. In all of the above cases the layers are in equilibrium insofar as the layers can remain in contact indefi- nitely, without changing their concentrations, so long as the tem- perature and other conditions remain constant. Whatmough did not measure the surface tension of one layer against the other; this was done by Antonow''^