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.

     

Refined liver extract in tropical macrocytic anemia

1. A series of 45 cases of T.M.A. treated with "refined" liver extract is reported.

2. "Refined" liver extract was found to be effective in 39 cases.

3. It was found that 2 or 3 ml. of refined liver extract (Examen N.P.) was sufficient to produce an optimum response.

4. As judged from therapeutic observations, it is suggested that in the majorityof cases of T.M.A. the deficiency is similar to that in Addisonian pernicious anemia, though the mode of production of the deficiency may not be the same.

Note: ACKNOWLEDGMENTOur thanks are due to Dr. R. Row, Hon. Director, P.G. Singhance Hindu Hospital, Bombay, whereall the cases were treated, for permission to publish these reports, and the firms concerned for the generousgift of liver extracts.

     

MONOCYTIC LEUKEMIA AND TUBERCULOSIS

A case of monocytic leukemia associated with active tuberculosis of pleura,pericardium, and lymph nodes is reported.

The criteria for the diagnosis of monocytic leukemia and the factors excludinga leukemoid reaction are presented.

The rarity of monocytic leukemia in contrast with the frequency of tuberculousinfections and the rarity of active tuberculosis in the reported cases of monocyticleukemia indicate that the association of the two diseases is probably coincidental.

Note: Grateful acknowledgment is due to Miss Anne Shiras and Mortimer Cohen, M.D., for the photo-micrographs.

     

A study of sickling of young erythrocytes in sickle cell anemia

1. Data have been presented to show that most reticulocytes from patients withthe sickle cell trait or sickle cell anemia sickle as readily as do more mature redblood cells. The most immature reticulocytes and normoblasts tend to sickle moreslowly.

2. Orthochromatic normoblasts were the only type of normoblasts whichsickled; the basophilic and polychromatophilic types could not be sickled.

3. It is suggested that the sickle cell forms seen in ordinary stained smears represent old cells which have lost their "elasticity" while stagnating in the sickleshape, and are unable to revert to a biconcave disk. This would explain the factthat these forms are so rarely found to be reticulated when stained with brilliantcresyl blue.

Note: ACKNOWLEDGMENTI wish to express my gratitude to Dr. William B. Castle, Dr. William Dock (Department of Medicine)for many stimulating suggestions, to Dr. John M. Pearce (Department of Pathology) for assistance inpreparing this report and to Mrs. Muriel MacDowell (Department of Pathology) for the photomicrography.

     

Preservation of normal human plasma in the liquid state; clinical,chemical, and physicochemical studies during three years of storage at room temperature

1. During preservation of human plasma in the liquid state at room temperaturefor three years, the alpha-amino nitrogen and the nitrogen content of the tungsticacid filtrate rise slowly until about two years have elapsed, after which time theselevels do not increase significantly.

2. During the entire period of storage the nitrogen contents of the trichlor-acetic acid filtrate and the "polypeptide index" increase progressively. Theactual increase represents hydrolysis to nonprotein size of 3 to 4 per cent of theoriginal protein.

3. The colloid osmotic pressure of stored plasma is slightly but significantlygreater than that of fresh plasma.

4. The viscosity of stored plasma is slightly but significantly greater than thatof fresh plasma.

5. The electrophoretic patterns of stored plasma show increases of alpha globulin and albumin concentration, complete disappearance of gamma globulin(containing immune properties) and fibrinogen, and some reduction of beta globulin concentration as compared to fresh plasma.

6. Analysis of 3,384 questionnaires completed after administration of liquidplasma more than a year old indicates that the transfusion of such plasma continues to be safe and beneficial up to at least three years of storage. The untowardreaction rate following these administrations was significantly less than thatfollowing a comparable series of 1500 administrations of commercially prepareddried plasma.

Note: The technical assistance of R. L. Erickson, PhM1c, V6, U.S.N.R., and P. Livingood, PhM3c, V6, U.S.N.R., is gratefully acknowledged. The kindness of Dr.Ellice McDonald, Director, Biochemical Research Foundation, in arranging for theelectrophoretic work and in permitting the reporting of the work here is appreciated with deep thanks.This study was possible only with the close cooperation of the Blood and PlasmaDepartment, Naval Medical School. For this cooperation, the authors are considerably indebted to Commander S. T. Gibson, M.C., U.S.N.R., LieutenantCommander H. R. Evans, (HC), U.S.N., and Lieutenant Commander M. T. Sproul,H(W), U.S.N.R.

     

THE TREATMENT OF LYMPHOBLASTIC LEUKEMIA WITH CRUDE MYELOKENTRIC ACID

Eight cases of blastic lymphoid leukemia have been treated with myelokentricacid in crude form, because hypothetically in blastic lymphoid leukemia there isa deficiency of this material. The crude myelokentric acid was used because it wasmore easily obtained than partially purified material. Purification of biologicallyactive materials by methods of extraction and precipitation necessarily results in aconsiderable loss of material. Thirteen partial remissions occurred following theadministration of crude myelokentric acid. Seven of the 8 patients have died, and5 necropsies were performed.

The necropsy material adds further weight to the belief that the remissions wereinduced by the myelokentric acid in that in all 5 necropsies there was a definitealteration in the histologic morphology as contrasted with the findings in thenecropsies of the controls.

It seems inadvisable, however, to treat a large number of patients with this material because it is crude, it is relatively unavailable, and no standard dose has yetbeen devised.

Note: We wish to t hank Dr. D. L. Turner and Dr. W. A. Hause for valuable assistance in this work.

     

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.

     

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 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     

Changes in circulating leukocytes induced by the administration of pituitary adrenocorticotrophic hormone in man

1. Pituitary adrenocorticotrophic hormone (ACTH), when administered in asingle intramuscular dose of 25 mg. to human subjects with unimpaired adrenalfunction, results in a characteristic alteration of the leukocytic pattern. This consists of an increase of circulating neutrophils and a decrease of circulating lymphocytes and eosinophils.

2. The decrease in circulating lymphocytes and eosinophils is contingent uponthe stimulation of a functionally competent adrenal cortex, and does not occur inits absence. The neutrophilic response is present but somewhat diminished inadrenal insufficiency.

3. The entire pattern of leukocytic alterations found in normal subjects after administration of ACTH can be induced in patients with Addison’s disease by 17-hydroxycorticosterone (20 mg.) but not with desoxycorticosterone glucoside (30mg.).

4. Prolonged adrenal stimulation by ACTH, given over a four day period in adose of 10 mg. every six hours, results in a sustained and striking elevation ofneutrophils and depression of eosinophils; the lymphocytes, after an initial depression lasting not more than twenty-four hours, may increase above their initial levels in spite of the continued increased secretion of adrenal hormones.

5. The relation of the adrenal cortex to the characteristic nonspecific leukocytepattern, observed as a response of the organism to any type of insult, is discussed.

Note: ACKNOWLEDGEMENTSWe are indebted to Mrs. Elizabeth Dell, B.S., Mrs. Constance Forsham, B.S., M.T. (A.S.C.P.), andMiss Loretta Conroy, B.S., for their technical assistance, and to Miss Mary Kascht, B.A., Research Dietitian, who kindly took over the secretarial management in addition to the dietary work.

     

Eosinophilic leukemia; report of a case with autopsy confirmation; review of the literature

1. The data in a case of fatal leukemia with predominant eosinophilia in theperipheral blood and bone marrow are presented; we believe that this case was oneof eosinophilic leukemia.

2. During the period of observation, these eosinophils showed progesssiveimmaturity as the symptoms became more severe. Eventually this "left shift"became so marked that a large proportion of the cells were terminally myeloblastsin both the blood and the bone marrow.

3. Autopsy revealed invasion of many of the tissues and organs with thesemature and immature eosinophil granulocytes and with myeloblasts.

Note: ACKNOWLEDGMENTOur thanks for helpful criticism are particularly extended to Dr. Charles Doan and to Dr. WilliamDameshek.

     

Dangerous universal donors; II. Further observations on in vivo and in vitro behavior of isoantibodies of immune type present in group O blood

1. Severe hemolytic reactions were observed in 3 group A (subgroup A₁) recipients transfused with group O whole blood or plasma. In one case, 10 ml. of acommerical preparation of soluble A and B factors had been added to 500 ml. ofwhole blood prior to the transfusion and it is believed that the reaction mighthave been even more serious had not this material been added.

2. The anti-A antibodies in the serum of the dangerous universal donors causingthe hemolytic reactions fixed complement, acted as hemolysins, were difficult toneutralize with soluble A and B factors, were capable of giving positive Coombstests and their ability to agglutinate A cells was enhanced by the presence of normal human serum. These characteristics were similar to those observed in serumfrom donors known to be actively immunized against the A factor, but the stimulusfor development of "immune" anti-A antibodies in the dangerous group O donorswas not apparent.

3. Small amounts of immune A antibody were consistently demonstrated in 12of 100 random group O sera which, after neutralization, produced indirect Coombstests with A₁ cells and agglutinated A₁ cells suspended in compatible normal humanserum.

4. Screening procedures for elimination of dangerous group O donors are discussed.

Note: ACKNOWLEDGMENTSIt is a pleasure to thank Mrs. Jane Peters, Mrs. Nieves Dole and Miss Jean Dorothy for technicalassistance and Dr. R. Wendell Davis for collaboration in the study of Case 1.

     

Refractory megaloblastic anemia

1. Fifty-nine cases of megaloblastic anemia refractory partially or completelyto potent liver extracts given parenterally have been investigated in Edinburghduring the past six years. Thirty-four of these cases were associated with pregnancy, the puerperium or the sprue syndrome. No explanation of the cause of themegaloblastic anemia was discovered in the remaining 25 cases.

2. The etiology, clinical features and treatment of 25 cases of idiopathic refractory megaloblastic anemia are described. Attention is directed to the excellenttherapeutic effects produced by proteolysed liver or folic acid.

3. The mechanisms involved in refractoriness to potent parenteral liver extractsare discussed.

4. In certain cases of refractory megaloblastic anemia it is suggested that an unknown hematinic principle, in addition to the liberating factor in purified parenteral liver extract and folic acid, is required for the complete restoration of normoblastic blood formation.

Note: ACKNOWLEDGMENTSMy thanks are due to many members of my staff who have helped in theseinvestigations, particularly to Professor L. J. Davis formerly lecturer in Medicinein the University of Edinburgh, and to Dr. Girdwood. Grateful acknowledgmentmust also be made to Doctor Riding, Medical Director of Evans Medical SuppliesLtd. and his research chemists who were responsible for the preparation of proteolysed liver and the other fractions of liver mentioned above.

     

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.

     

Studies on Charcot-Leyden crystals

1. Synthetic detergents of the anionic, cationic and nonionic types result inthe rapid and constant formation of Charcot-Leyden crystals from eosinophils.

2. Charcot-Leyden crystals have a negative crystalline birefringence and formpenetration twins.

3. The changes taking place in the eosinophil in the formation of Charcot-Leyden crystals under the influence of wetting agents, utilizing phase and polarizing microscopy, are described.

4. In the formation of Charcot-Leyden crystals with wetting agents, the nucleusof the eosinophil lyses with no appreciable effect on the granules.

5. In the formation of Charcot-Leyden crystals with a wetting agent, there isno change in the lipoid cortex of the eosinophil as demonstrated by staining withsudan black B.

6. Charcot-Leyden crystals undergo changes on standing that affect theirsolubilities.

7. The staining reactions and solubilities of Charcot-Leyden crystals aredescribed.

8. Oxyhemoglobin crystals constantly form from red cells on exposure to Aerosol MA; on two occasions, tyrosine crystals formed from the blood of a patientwith chronic myelogenous leukemia.

9. Evidence is offered that Charcot-Leyden crystals are crystalline proteinsderived only from the nucleus of the eosinophil.

Note: ACKNOWLEDGMENTSI wish to thank Dr. William B. Ober, Dr. J. J. Englefried, and Lt. E. E. Ozburn, MSC, USN, forassistance in this work.

     

Folic acid antagonists in the treatment of acute and subacute leukemia

Forty cases of acute and subacute leukemia were treated with one or more ofvarious folic acid antagonists, usually aminopterin. Thirty-two cases were treatedfor at least one week, remissions occurring in 10. The remissions were of a temporary nature and variable in duration. As of June 1, 1949, two patients werestill alive and in good condition, seven and one-half and thirteen months, respectively, after onset of therapy.

Clinical, hematologic, and to lesser extent, marrow remissions were obtainedmost commonly in the lymphocytic type, none in the 4 monocytic cases. The subacute cases responded far better than did those of the acute, fulminating variety.

The exact mechanism of action of the folic acid antagonists is not known, although one may speculate that anti-PGA, an anti-growth factor which resemblesPGA so closely, is readily accepted by the primitive white cell with resultantcell death.

The margin between a therapeutic or effective dose and one causing a toxic reaction is a narrow one. A "toxic" reaction may in fact be an indication of a therapeutic response.

The pattern of therapy was (1) administration of the drug, usually by parenteralinjection until a toxic or a pronounced hematologic reaction occurred, at whichpoint (2) the drug was discontinued. With subsidence of the toxic reaction, (3)a maintenance dosage was then given, usually in the form of oral medications,0.5 mg. daily for adults and 0.25 mg. for children.

Transfusions and antibiotics were administered as indicated. Frequent transfusions were usually very helpful at the time of the initial reaction when anemiawas ordinarily severe. Penicillin and other antibiotics were used for supportivetherapy. Thrombocytopenia and hemorrhage were exceedingly difficult to control.

The observed remissions appeared to be directly attributable to the action ofthe drug. Although temporary, they indicate that acute leukemia is not necessarily completely irreversible. Thus, there is hope that more potent anti-growthfactors may someday be discovered which will be of value in ultimate control ofthe disease.

Note: ACKNOWLEDGMENTSWe are pleased to acknowledge the continued cooperation and generosity of various members of theStaff of Lederle Laboratories. We wish particularly to mention the help of the late Doctor YellapragadaSubbaRow, who was greatly interested in some of our first remissions obtained in adults. We also wish tothank the staffs of the U. S. Veterans Hospital at West Roxbury (Dr. Thomas Warthin, Physician-in-Chief) and of the Murphy General Hospital for their cooperation. We are greatly indebted for their helpto the medical and nursing staffs of the Boston Floating Hospital (Dr. James H. Baty, Physician-in-Chief) and of the New England Center Hospital (Dr. Samuel Proger, Physician-in-Chief).

     

The relationship between carbonic anhydrase activity and zinc content of erythrocytes in normal,in anemic and other pathologic conditions

A good correlation exists between zinc content and carbonic anhydrase activityof the red blood cells under all conditions studied, including anemia and polycythemia. In almost all patients with anemias other than pernicious anemia, bothzinc and carbonic anhydrase levels were lowered in parallel fashion. These changeswere proportional to decreases in hematocrit and hemoglobin levels and erythrocyte counts so that both zinc and carbonic anhydrase values per unit of RBC werein the normal range. In a few instances of anemia associated with leukemia andin one of sickle cell anemia, neither zinc content nor carbonic anhydrase activitywas decreased in proportion to the anemia; in these cases the zinc and carbonicanhydrase levels per unit of blood were both elevated to the same degree.

Patients with pernicious anemia showed no decrease in absolute values for zincand carbonic anhydrase activity in spite of marked lowering of hematocrit andhemoglobin levels and of erythrocyte count. Accordingly, both zinc concentrationand carbonic anhydrase activity per unit of blood were elevated, often to a markeddegree. These increases were parallel, varying inversely with the degree of anemia;when they regressed under treatment, both did so at the same rate.

There are no methods available for estimating carbonic anhydrase concentration;all methods now in use measure only the activity of the enzyme. It is suggestedthat zinc concentration could be used as an indicator of carbonic anhydrase contentof the red blood cells.

Note: ACKNOWLEDGMENTSDrs. Joseph C. Aub and Ira T. Nathanson were kind enough to refer several patients for study. Dr.Byrl J. Kennedy was most helpful in regard to obtaining samples of blood. The technical work wasperformed by Miss Mary Lou Roney, Betty Hickey and Marion Taylor.

     

Inconstancy and variability of the vascular fragility test even in purpuric conditions

Two unusual cases of purpura simplex raise the problem, as yet unsolved, of thepathogenesis of the purpuric eruption. Together with a previous observation ofhemorrhagic purpura made by Roskam, they show the inconstancy and variabilityof the vascular fragility test.

Note: ACKNOWLEDGMENTThis paper was translated by Raymond G. Gottschalk, M.D., whose cooperation is gratefully acknowledged.

     

The Rh chromosome frequencies in England

The results are reported of testing 1073 English bloods with the Rh antibodiesanti-C, anti-C^w, anti-c, anti-D, anti-E and anti-e. The results of another series of927 bloods, already published, are here reproduced. The total of 2000 bloods hasbeen used by Fisher to estimate, by his method of maximum likelihood, the Rhchromosome frequencies in England. The estimates are: CDe 40.75 per cent, cde38.86 per cent, cDE 14.11 per cent, cDe 2.57 percent, C^wDe 1.29 per cent, cdE 1.19per cent, Cde 0.98 per cent, and CDE 0.24 per cent.

A brief account is given of the three pairs of alternative antigens shown byFisher to be the basis of the Rh blood groups. Fisher’s interpretation must now beconsidered as established beyond doubt. A possible genetic basis of these relatedantigens is discussed.

Note: ACKNOWLEDGMENTSWe are deeply indebted to Professor Fisher for many reasons, but we should particularly like toacknowledge his kindness in allowing us to publish the results of his calculations of the chromosomefrequencies.For the antisera used in the investigations we are indebted to the following: Doctors E. F. Aubert,Sheila Callender, D. S. Dick, R. J. Drummond, Mr. I. Dunsford, Doctors A. J. McCall, Brenda Morrison,J. Murray, E. Wordley and R. A. Zeitlin.We also thank Dr. H. F. Brewer of the London Red Cross Blood Donor Service, and Dr. J. F. Loutit ofthe National Transfusion Service, for providing us with large numbers of blood samples.We also wish to acknowledge the assistance of Dr. Marjory N. McFarlane in the earlier part of thisinvestigation.

     

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.