Experimental anemias in the rat; I. Macrocytic anemia in chronic pteroylglutamic acid deficiency and after splenectomy in Bartonella muris infection

1. In agreement with findings by other workers, rats in acute pteroylglutamicacid deficiency showed leukopenia and growth depression followed by death, without any significant change in the red cell picture.

2. In chronic deficiency, however, produced by the addition of small pteroylglutamic acid doses given intermittently, a severe anemia was obtained afterseventy days.

3. The anemia was macrocytic and "normochromic." Price-Jones curves showeda preponderance of macrocytes with anisocytosis. This agreed with findings byother workers for other species.

4. The anemia could be cured by single doses of 40 µg. or more of pteroylglutamicacid.

5. There was no significant difference between sexes to pteroylglutamic aciddeficiency. Reduction in the protein content of the diets, containing 1 per centsulfasuxidine, from 18 per cent to 10.5 per cent, produced no significant differencein the time of onset and severity of the blood symptoms.

6. These results were not due to infection with Bartonella muris. This infectionproduced a macrocytic anemia of a different type, and was curable by treatmentwith neoarsphenamine.

Note: ACKNOWLEDGMENTSWe are grateful to Dr. T. H. Jukes of the Lederle Laboratories for generous supplies of aldehyde-freePGA; and to Dr. K. Folkers of Merck Laboratories for the biotin used in these experiments. We wish tothank Dr. W. Jacobson for his advice during the course of this investigation. Valuable technical helpwas provided by Mr. D. R. Ashby, Mr. S. G. Impey, Miss M. J. Kemp and Mr. P. W. Wilson, to whomthe authors are indebted.

     

A SIMPLE APPARATUS AND PROCEDURE FOR PREPARING AND ELECTROPLATING RADIOACTIVE IRON

A simple, easily constructed apparatus for electroplating radioactive iron hasbeen described by which certain disadvantages of other forms of apparatus havebeen eliminated. Sixteen samples can be plated each day without difficulty.

The technic employed for preparing the radioactive iron for electroplating hasbeen given.

Note: The writers are indebted to Dr. P. F. Hahn and his staff who permitted one of them (M. L.) to spendseveral weeks in the Biochemical Laboratory at the Vanderbilt University School of Medicine and generously gave invaluable instruction and advice.

     

Autohemagglutinins and hemolysins with hemoglobinuria and acute hemolytic anemia,in an illness resembling infectious mononucleosis

A case is reported of a young man with acute hemolytic anemia and hemoglobinuria who presented: an initial blood picture consistent with infectious mononucleosis, associated with a heterophile agglutination test positive in high dilution;auto-hemagglutinins, active in the cold, at room temperature and at 37 Centigrade;a hemolysin active at 37 C. after chilling, requiring the presence of a thermolabilecomponent of serum for hemolysis; a positive Donath-Landsteiner test but no evidence of syphilis. In addition there was clubbing of the digits with certain otherroentgenologic changes in the bones; absence of any other etiologic factors knownto be concerned with such anemia; uneventful improvement under massive transfusion therapy, with apparent recovery from his hematologic disorder whenstudied two years later.

Note: ACKNOWLEDGMENTOur thanks are due to Dr. Maxwell Finland and Dr. Philip F. Wagley for suggestions in preparingthis report. We are especially indebted to Dr. T. Hale Ham for his patient and critical advice and encouragement.

     

Tuberculous splenomegaly with the hypersplenism syndrome; a case report

A case of tuberculous splenomegaly with leukopenia and anemia followingmiliary tuberculosis has been presented. Splenectomy was required after streptomycin failed to control the cytopenias, progressive emaciation, and splenic infection. However, following what appeared to be six weeks of marked improvement,the patient developed a fulminating tuberculous meningitis and died.

Note: ACKNOWLEDGMENTThe authors are indebted to Dr. Byrd S. Leavell for his suggestions in the preparation of this paper.

     

Experimental production of a nutritional macrocytic anemia in swine

1. A deficiency of pteroylglutamic acid has been produced in 32 swine fed apurified diet containing casein and supplemented with seven B vitamins, sulfasuxidine and a folic acid antagonist. The casein was fed at two levels, 10 and 26per cent. Two types of casein were used: a crude preparation possessing significant"extrinsic factor" activity and a purified casein with little activity.

2. The hematologic manifestations observed were (a) severe macrocytic anemia,(b) leukopenia, due to a proportionately greater reduction in polymorphonuclearthan in mononuclear cells, (3) slight thrombocytopenia, and (4) hyperplastic bonemarrow with an increase in immature nucleated red cells which resemble themegaloblasts seen in the bone marrow of patients with pernicious anemia.

3. The feeding of a 26 per cent rather than a 10 per cent crude casein diet did notprevent but did delay the onset of the blood changes. Anemia developed mostrapidly in the animals receiving 10 per cent purified casein.

4. The group receiving 26 per cent casein developed a greater degree of macrocytosis in the same period of time than did the group receiving 10 per cent casein.In all groups the degree of macrocytosis increased as the duration of the anemiaincreased.

5. The hematologic manifestations were not delayed nor was their developmentprevented by the intramuscular administration of 15 U.S.P. units of liver extractevery 15 days.

6. The blood and bone marrow returned rapidly to normal following the administration of pteroylglutamic acid, pteroyldiglutamic acid, pteroyltriglutamicand pteroylheptaglutamic acid. Thymine and xanthopterin had little or no activity. Tyrosine, adenine and uracil were inactive.

7. Purified liver extracts and crystalline vitamin B₁₂ were found to possess somehemopoietic activity in several animals but the activity was considerably less thanthat of the pteroylglutamic acid compounds.

8. The urinary excretion of "tyrosyl" (hydroxphenyl compounds) was notabnormal in the pteroylglutamic acid deficient pigs and was not altered by eitherpteroylglutamic acid or liver extract therapy.

9. The urinary excretion of allantoin and uric acid did not differ significantlyfrom the normal. Immediately following therapy with pteroylglutamic acid,however, in association with the reticulocytosis and lasting for the same period,there was a marked increase in the excretion of allantoin.

10. The results suggest that both pteroylglutamic acid and a factor in liverextract similar to or identical with vitamin B₁₂ are required for normal hemopoiesisin the pig.

Note: ACKNOWLEDGEMENTSThe crude methylfolic acid antagonist, xanthopterin, and the pteroylglutamic acid compounds, withthe exception of pteroylheptaglutamic acid, were kindly furnished by the Lederle Laboratories, PearlRiver, New York, through the courtesy of Dr. T. H. Jukes and Dr. S. M. Hardy.Sulfasuxidine was generously furnished by Sharp & Dohme, Inc., Philadelphia, Pa., through thecourtesy of Dr. W. A. Feirer.Pteroylheptaglutamic acid and Natola were supplied by Parke, Davis & Company, Detroit, Mich.,through the courtesy of Dr. A. E. Sharp and Dr. J. J. Pfiffner.Biotin was obtained from Hoffmann-LaRoche, Inc., Nutley, N. J., through the courtesy of Dr. E. L.Sevringhaus.The vitamins, with the exception of pteroylglutamic acid and biotin and including vitamin B₁₂ werekindly furnished by Merck and Company, Inc., Rahway, N. J., through the courtesy of Dr. A. Gibsonand the late Dr. D. F. Robertson.Experimental liver extracts (No. 1124, 1063, 1066 and 1067) were generously furnished by Armour andCompany, Chicago, Illinois through the courtesy of Dr. E. E. Hays.We are indebted to Mrs. Darlene Kehl, Mr. George Trappett, and Mr. Ocie Hadley for technicalassistance.

     

Granulomatous lesions in the bone marrow in infectious mononucleosis; a comparison of the changes in the bone marrow in infectious mononucleosis with those in brucellosis,tuberculosis, sarcoidosis and lymphatic leukemia

1. The findings in the blood and in aspirated bone marrow in 23 cases of infectious mononucleosis have been described.

2. Unequivocal evidence of involvement of the bone marrow has been found in70 per cent of the cases.

3. Evidence of granulomatous inflammation of the marrow was found in 48 percent of the cases.

4. Epithelioid cells were found in the films of bone marrow in 48 per cent of thecases. These cells appear morphologically identical with those seen in imprints oflymph nodes from infectious mononucleosis and sarcoidosis and with the epithelioid cells seen in films of the marrow in brucellosis, sarcoidosis and tuberculosis.

5. The granulomatous lesions of infectious mononucleosis seem most similar tothose of brucellosis, but they also resemble the small granulomatous lesions ofsarcoidosis and tuberculosis.

6. Lymphocytosis of the marrow as well as of the blood was demonstrated in allcases. Evidence of formation of lymphocytes in the marrow was presented, and thealtered lymphocytes of infectious mononucleosis were found in films of the marrow.The degree of lymphocytosis of the marrow in infectious mononucleosis was shownto be less than that in lymphatic leukemia. Lymphocytosis of the marrow was notfound in brucellosis, sarcoidosis or tuberculosis. The lymphocytic reaction demonstrable in the marrow in infectious mononucleosis is believed to be of value in differential diagnosis.

Note: ACKNOWLEDGMENTSWe wish to acknowledge the generous cooperation of Dr. Ruth E. Boynton and the Staff of theStudent’s Health Service of the University of Minnesota throughout the course of this year long studyof infectious mononucleosis. We are indebted to Dr. T. Edward Bell and Dr. James Cardy for performingthe sternal aspirations. The photomicrographs were made by Mr. Henry Morris.

     

A histochemical study of acid and alkaline phosphatase distribution in normal bone marrow smears

1. Three minute fixation in formol vapor at 44 C, followed by 15 minute washing proved to be the most satisfactory fixation procedure for both "acid" and "alkaline" phosphatase technics as applied to bone marrow smears.

2. For both technics a relation between staining intensity and cellular richnesswas found.

3. The reaction of normal human bone marrow cells to both phosphatase technics is described. Both are predominantly nuclear in location. Nuclear patternapproached that observed with common staining methods and Feulgen’s reaction.Cytoplasmic reaction was nearly negative. Nonspecific and specific granules donot stain after the "alkaline" technic. Nonspecific granules are negative for "acid"phosphatase, while specific neutrophilic are variable, and eosinophilic, constantlypositive. Nucleoli are negative after the "acid" technic, being positive for the"alkaline" enzyme. Mitotic chromosomes are positive for both technics. "Acid"phosphatase reaction in cytoplasmic zones of lymphocytes, erythroblasts, plasmacytes and megakaryocytes, is described.

Note: ACKNOWLEDGMENTSMany points pertaining to this paper were discussed with the late Dr. José Oria, to whom we oweinvaluable stimulation and guidance. We are indebted to Prof. W. Buno of Montevideo (Uruguay) forkind advice on the Golgi apparatus of blood cells. We are deeply indebted to Professor O. P. Jones ofBuffalo for his painstaking help and advice. We thank Dr. M. A. Jamra (Section of Hematology, CentralLaboratory, Hospital das Clinicas da Faculdade de Medicina) and Dr. J. F. Pontes (Section of ClinicalTherapeutics, Hospital das Clinicas da Faculdade de Medicina), for part of the material used in thisstudy. We thank Messrs. L. Frankenthal and G. Lonenzini for their aid in the preparation of some of themicrophotographs.

     

SERUM PROTEIN CHANGES IN MYELOGENOUS AND LYMPHOCYTIC LEUKEMIAS AND HODGKIN'S DISEASE

1. Using a methyl alcohol fractionation technic, the albumin, globulin, andtotal protein levels were determined in a series of normal adults and compared withcases of myelogenous and lymphocytic leukemias and Hodgkin’s disease.

2. Statistically significant decreases in albumin and increases in globulin werefound in the cases of Hodgkin’s disease and myelogenous leukemia, but withoutsignificant changes in total protein. Globulin levels above the highest normal valuewere found in 23 per cent of the former and 33 per cent of the latter group.

3. No apparent relationship was noted between the levels of the serum proteinfractions and (1) the hemoglobin level, (2) the erythrocyte count, (3) the peripheral white blood cell picture, and (4) the bone marrow smears.

Note: The authors wish to express their appreciation to Professor Ovid O. Meyer, Department of Medicine,for making available the clinical material in this study and for valuable suggestions. The authors arealso indebted to Professor J. A. E. Eyster, Department of Physiology, for suggestions as to statisticaltreatment of the data.

     

The mechanism of petechial hemorrhage formation

1. The capillary resistance test has been studied by a special technic of capillarymicroscopy.

2. Seventeen cases of hemorrhagic diseases of differing etiology have been thusstudied.

3. The site of capillary hemorrhage has been localized to the arteriolar end ofthe capillary loop.

4. Selective damage to the arteriolar-capillary junction will explain manytypes of hemorrhagic syndromes.

Note: ACKNOWLEDGMENTI wish to thank the honorary staff of the Westminster Hospital for permission to study cases undertheir care. I am much indebted to Dr. P. Hansell and Miss F. E. McAdoo of the Westminster HospitalDepartment of Medical Photography for assistance with the illustrations.

     

Microscopic and histochemical studies on the Auer bodies in leukemic cells

(1) Seventeen cases of acute leukemia with Auer bodies have been reported.Studies were carried out on 7 cases of acute monocytic leukemia and 3 cases ofsubacute myelogenous leukemia.

(2) Histochemical studies showed the Auer bodies to be oxidase, peroxidase,and periodic acid-Schiff positive; sudanophilic, slightly metachromatic and to givepositive tests for acetal lipids and ribonucleic acid.

(3) The Auer bodies were negative for acid and alkaline phosphatase, lipase,glycogen, desoxyribonucleic acid and were non-birefringent.

(4) A change in the chemical nature of the Auer body from an acid conditionto a more neutral state was noted. This change corresponded with the changesof the normal cytoplasmic granulation of the myelocytes and monocytes duringmaturation.

(5) The effects of cellular movements, trauma, and temperature changes uponthe Auer bodies were studied.

(6) Several leukemic cells, containing Auer bodies, were studied during theprocess of mitosis.

(7) A theory as to the formation and disintegration of the Auer bodies hasbeen presented.

Note: ACKNOWLEDGMENTSThe author wishes to express his thanks to Dr. B. K. Wiseman, Dr. B. C. Houghton, Dr. R. A. Knouff,and Dr. E. R. Hayes for their help and suggestions, and to Dr. J. D. Thomas, Dr.J. F. Gamble, Dr. R. J.Rohn, Dr. H. Schiro, Mrs. Jo Myers, M.Sc., Miss Georgia Gwinner, M.T., Mrs. Susan Ragsdale, M.T.and Mrs. Jeanne Marie Willison, M.T., for their help in securing the experimental material.

     

The Lutheran blood groups: a second example of anti-Lu b and three further examples of anti-Lu a

1. An example of the blood group antibody, anti-Lu^b, was found in a patientwho had a mild hemolytic transfusion reaction. It was shown to possess thecharacteristics of an immune antibody and to be able to distinguish between asingle dose and a double dose of the Lu^b gene.

2. Three new examples of the antibody, anti-Lu^a, are presented. All of themwere found in normal blood donors and have properties which indicate that theyare naturally occurring antibodies.

Dr. R. R. Race and Dr. R. Sanger confirmed the presence of anti-Lu^b in Mrs. S.’sserum, and studied other members of her family and the three anti-Lu^a sera. We aregrateful to them for many favors and their kind encouragement.

We are obligated to Miss Marie Cutbush for making available the Lu^aLu^a cells fromMrs. R. and her sister, and for a supply of anti-Lu^b serum.

Thanks are due to Dr. A. E. Mourant who furnished our original supply of anti-Lu^aserum and to Dr. Philip Levine for the anti-Tj^a and anti-Vel sera.

We are indebted to Dr. J. M. Fine of Milwaukee for permission to study Mrs. S. andto the patient and her family for their cooperation.

The sera from 18,613 blood donors were studied by Betty McCarthy, Rosemary Polka,Pearl Lemke, Agnes Molnar, Jeannette Flagstadt and Betty Hutter.

Submitted on March 20, 1957 Accepted on July 1, 1957     

Experimental anemias in the rat; II. The effect of various sulfonamides in producing a pteroylglutamic acid deficiency and the pteroylglutamic acid activity of test substances

1. Different sulfonamides were tested to ascertain their effect in producing thecharacteristic symptoms of acute PGA deficiency in rats fed on synthetic diets.Sulfasuxidine (1 per cent) and the less soluble phthalylsulfathiazole (1 per cent)were equally effective. Sulfathiazole in 1 per cent concentration produced a hemolytic anemia not reversible by PGA or whole liver powder. In a 0.5 per cent concentration it was also effective, but in view of its toxicity, the less soluble sulfonamides were to be preferred. A mixture of 0.5 per cent sulfathiazole and 0.5 percent sulfadiazine was extremely toxic and produced a hemolytic anemia. Sulfaguanidine was toxic at 1 per cent concentration.

2. When intermittent small doses of PGA were given to PGA-deficient rats toprolong their life from 45 up to 155 days, 1 per cent sulfasuxidine or phthalylsulfathiazole, or 0.5 per cent sulfathiazole were equally efficient in producing regularlya macrocytic normochromic anemia.

3. The response of PGA-deficient rats to single doses of PGA has been studiedand an assay procedure has been suggested which uses the weight increase andduration of cure as the measure of the response. The W.B.C. and reticulocyte response can also be used as a qualitative indication of PGA activity.

4. Of the substances tested by this procedure, vitamin B₁₂, purified perniciousanemia preparations, ascorbic acid and xanthopterin showed no PGA activity. Acommercial yeast preparation and Teropterin were found to possess biologic activity comparable with that found by other workers in assays on chicks.

Note: ACKNOWLEDGMENTSWe wish to thank Dr. E. Lester-Smith, of the Glaxo Laboratories, for the generous supply of vitaminB₁₂, Dr. T. H. Jukes, of the Lederle Laboratories, for the gift of PGA and Teropterin, and Dr. W. Jacobson for the samples of purified P. A. liver preparations and xanthopterin.

     

Hemoglobin levels among 7 to 14 year old children in Saskatoon,Canada

In May and June of 1948 a survey of hemoglobin values was carried out on 842girls and 827 boys, ages 7 tO 14 inclusive, in Saskatoon public schools, who represented about a third of this population group in the city.

Values for girls ranged from 10.0 to 16.3 Gm. per 100 ml. of blood, with anaverage of 13.5. The corresponding values for boys were 11.2. tO 16.7, and 13.7.The average for the whole group was 13.6, which is among the highest even reported for children of these ages. The average values for children in the differentage groups were slightly higher than those reported for comparable groups in otherparts of Canada.

Values for boys and girls of the same age were similar. There was an increasefrom near 13 in the 7 year old group to near 14 in the 14 year old group. The similarity in hemoglobin values for both boys and girls of these ages, and the small increase from 7 tO 14 years of age, are in agreement with observations of otherworkers.

Eleven per cent of the children had been subject to more than the usual degree ofillness. No significant differences appear when the above figures are corrected byremoving this group.

Note: ACKNOWLEDGEMENTSWe are greatly indebted to Dr. Griffith Binning for his help and cooperation, without which thissurvey could not have been carried out. His interest and advice, not only during the course of the work,but in the preparation of this paper, were very much appreciated, and we should like to render him oursincere thanks.To the principals and staffs of the schools included in the survey we extend thanks for their courtesyand help.Dr. H. B. Collier, Head of the Department of Biochemistry, rendered valuable help and advice duringthe course of the survey, and in the preparation of this paper. We wish to acknowledge this with ourthanks.Helpful suggestions were received from Dr. L. B. Jaques, Professor of Physiology at the University ofSaskatchewan, and from Dr. E. W. McHenry, Professor of Public Health Nutrition at the University ofToronto. We extend to them our thanks.

     

STUDIES ON THE PHYSIOLOGY OF THE WHITE BLOOD CELL: THE GLYCOGEN CONTENT OF LEUKOCYTES IN LEUKEMIA AND POLYCYTHEMIA

The technic of determining glycogen in isolated white blood cells was appliedto the study of the different types of leukemia and of polycythemia, in order toobtain information on the physiology of the white blood cell. From this study itis concluded that the granulated leukocyte is the only carrier of glycogen in wholeblood. The "reducing substances" in lymphocytes and blast cells are not consideredas true glycogen.

The glycogen content of wet white blood cells in the rabbit amounts to about1 per cent. In the human being a range of from 0.17 to 0.67 per cent was calculated.In disease higher percentages occur, in polycythemia up to 1.64 per cent and inglycogen storage disease up to 3.05 per cent.

The glycogen concentration of normal white blood cells is within the same rangeas that of the striated muscle.

Note: I acknowledge with gratitude my indebtedness to Dr. William Dameshek for giving me the opportunity of analyzing the blood of some of the patients studied. Miss M. H. Campbell, Miss H. A. Clark,and Miss L. M. Garofalo have aided in carrying out many of the blood counts.

     

Quantitative Serologic and Isotopic Studies on the Rho Variant--Du

1. Type D^u cells take up less than 10 per cent of the I-131 anti-D boundto D positive cells, so that the quantity of cell bound I-131 did not differ significantly from that bound nonspecifically to D negative cells.

2. Papain modification of D^u cells results in an almost threefold increasein the uptake of I-131 anti-D, whereas, D negative cells show no consistentincrease after such treatment. Enzyme modification, therefore, serves to clearlydelineate the two cell types.

3. The range of uptake of I-131 anti-D to enzyme treated D^u cells is fromapproximately 7 to 17 per cent of that by R₁R₂ cells. Most of these results wereobtained from studies on Mendelian D^u types. Studies on several bloods frommembers of a family having Rh genotypes typical of the "gene interaction" D^uconfiguration (C in transposition to D) revealed no significant reduction inI-131 anti-D uptake when compared with D positive cells.

4. Decreased antibody binding to D^u cells was confirmed by the serologicmethods used. Absorption of an anti-D serum and antiglobulin inhibitionwith sensitized D^u cells indicated that the quantity of D antigen was lessthan that observed with the isotopic technic. Elution studies did not show asmarked a reduction of D antigen on the D^u cell as the other serologic technicsindicated. Possible reasons for these differences have been discussed.

5. Although, from both the serologic and isotopic studies, the data indicate that the type D^u red cell is quantitatively deficient in D antigen, theyare inadequate to rule out other explanations for their reduced D antigenactivity. Irrespective of the mechanism(s) responsible, the effective bindingof anti-D by D^u cells is reduced several fold; this would appear to be theprimary cause of the weak second stage of red cell antigen-antibody reactionwhich characterizes type D^u blood.

Submitted on July 28, 1964 Accepted on October 8, 1964     

Chronic marrow failure,myelosclerosis and extramedullary hematopoiesis

Thirty cases, including 20 autopsied, of chronic bone marrow failure, myelosclerosis and osteosclerosis have been presented and compared with similar reportsin the literature. The bone marrow histopathologic sequence observed involvedrepeated necrobiosis of maturing hematopoietic cells, followed by overgrowth ofmarrow reticulum and frequent ossification. Immature erythrocytes and leukocyteswere often found in the circulation, and extrameduallary hematopoiesis was characteristic. One case was complicated by leukemia. Etiologic factors implicatedincluded exogenous toxic chemicals, liver dysfunction, endocrine abnormalities,blood loss or destruction and cardiovascular disease. Suspicion was directed towardthe pathogenetic importance of protracted bone marrow exposure to certain substances normally conjugated rapidly in the liver and excreted.

Note: ACKNOWLEDGMENTThe authors acknowledge gratefully the continued assistance of Dr. John Norcross and Dr. DonatP. Cyr of Lahey Clinic, Boston, who permitted use of material from living cases.We are indebted to Dr. Shields Warren, New England Deaconess Hospital, Boston, for permissionto review 10 cases; to Dr. Olive Gates, Harvard Medical School, for advice and assistance with 7 cases,including material from the Collis P. Huntington Hospital; to Professor William Boyd for 5 cases fromBanting Institute, Toronto; to Dr. Paul D. Rosahn, New Britain General Hospital, New Britain, Conn.,and Dr. William Dameshek for one case; to Dr. A. J. Blanchard for several cases from Christie St. Hospital, Toronto; to Drs. Lorne Whittaker and Sabin of St. Catherine’s Hospital for one case; and to Dr.John Brownlee for hematologic studies from one case.

     

THE EXCRETION OF UROBILINOGEN IN THE STOOLS AND URINE DURING MALARIAL INFECTION

In 10 cases of malaria (6 benign tertian, 4 malignant tertian), the excretion ofurobilinogen in the stools and in the urine was studied. In all 10 cases the amountof urobilinogen excreted in the stools was found to be increased. After defervescence and disappearance of parasites from the blood the excretion gradually declined. The increased excretion of urobilinogen in the stools was the constant andsometimes the only evidence of increased blood destruction occurring at times in the complete absence of jaundice and reticulocytosis. Increased excretion of urobilinogenin the urine was not a constant feature.

It is suggested that the development of jaundice and of urobiligenuria is duenot only to the liberation of pigments by the hemolysis, but to a disturbance in theliver function.

This study lends further confirmation to the concept that the only unequivocalevidence of increased blood destruction is shown in an increased output of urobilinogen in the feces.

Note: The author is indebted to Dr. M. Rachmilewitz for his suggestions and criticisms.

     

The genetic relation and clinical differentiation of Cooley's anemia and Cooley's trait

1. Four additional families illustrating the clinical and genetic relationshipsof Cooley’s anemia and Cooley’s trait have been presented.

2. Blood findings in an offspring of a patient with Cooley’s anemia are recorded.

3. The asymptomatic nature of Cooley’s trait and its differentiation fromCooley’s anemia has been emphasized.

4. The inheritance of Cooley’s trait and Cooley’s anemia may be best explainedin terms of an incomplete dominant or of the simultaneous appearance of twononallelomorphic genes.

Note: ACKNOWLEDGMENTWe are indebted to Dr. John H. Linner for many of the observations on the Ca family and to MissClara Gillette for observations on the Cr family.

     

THE USE OF PHOTO-ELECTRIC TURBIDOMETRY IN THE DETERMINATION OF RED CELL COUNTS, HEMATOCRITS, AND HEMOGLOBIN

1. Blood turbidometry is recommended as a screening technic for distinguishingbetween anemic and nonanemic individuals.

2. Blood turbidometry must be supplemented by other technics for an exactdiagnosis of the type of anemia. However, with such help it makes its own contribution to the accuracy of the diagnosis.

3. Blood turbidometry alone would seem to be capable of following an anemicindividual’s response to therapy once the proper diagnosis is established.

4. Determination of hemoglobin concentration by turbidometry appears as adistinct possibility. However, further investigation is necessary to validate itsutility.

5. Evidence is accumulated that the shadow-volume relationship is a constantin several species.

Note: The author is particularly indebted to the following for advice in connection with this work: Dr.Peter Olafson of the Department of Veterinary Pathology, Dr. C. E. Hayden of the Department of Veterinary Physiology, Dr. L. I. Barnes of the Department of Physics, and Dr. W. B. Carver of the Department of Mathematics, all of Cornell University, and Dr. Charles P. Winsor of Johns Hopkins University.Dr. Robert N. Ericson conducted some preliminary explorations of the problem with the authorwhen both were connected with Kansas State College.

     

Response of tropical sprue to vitamin B12

These findings show that the administration of vitamin B₁₂ to patients with tropical sprue was followed by general clinical and hematologic improvement providedthe dosage was adequate. A single dose of 4 micrograms administered in case 4produced little or no change. The larger dosage of 10-25 micrograms administeredin the other cases was accompanied by striking increase in strength and vigor anda decrease in the diarrhea; however, in no instance was a maximal dose given andthese patients quickly tended to relapse clinically and hematologically. Theycould be relieved promptly again either by another injection of vitamin B₁₂ orby a compound of folic acid. (The conjugated compounds of folic acid used in thesecases were used for experimental purposes, and they produced the same hematologicresponse as that of folic acid per se.) Case 3, who had an excellent hematologicresponse after eating one serving of 400 grams of liver, is regarded as especiallysignificant in that it suggests that, as powerful as vitamin B₁₂ is as a therapeuticagent, it is more effective when given with liver. It is especially noteworthy thatcases 1 and 2, who had three injections of vitamin B₁₂, have had steady clinicaland hematologic improvement. The reader should have in mind that a singleinjection of approximately 100 micrograms of vitamin B₁₂ probably would beneeded to produce a full hematologic response in persons so ill. This tentative appraisal would suggest that this therapeutic compound, per unit of weight, is moreeffective in treating human disease than any compound that yet has been used.

Note: ACKNOWLEDGMENT We are very much indebted to others who have aided us in selecting cases and in observing results.Especially we wish to thank Dr. F. Hernandez-Morales, Dr. Hector Marchand, Miss Clemencia Benitez-Gautier, and Miss Sara Torres.