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.

     

THE EFFECT OF DIET ON THE HEMOGLOBIN, ERYTHROCYTE, AND LEUKOCYTE CONTENT OF THE BLOOD OF THE RHESUS MONKEY (MACACA MULATTA)

1. Rhesus monkeys fed purified rations supplemented with adequate amountsof the B vitamins, ascorbic acid, and whole liver substance maintained the following average blood picture:

See PDF for Table

2. Natural diets or purified rations supplemented with liver extract do not support the above blood picture. The hemoglobin is lower and there is an increase inthe range of the total leukocyte count and in the neutrophil-lymphocyte ratioto 2.0 ([unknown]). These figures are similar to the values in the literature and generallyaccepted as the normal.

3. Previous reports have shown the characteristic blood dyscrasias which develop when monkeys are fed certain B vitamin-deficient diets. These changes aresummarized graphically in this paper.

4. The importance of determining the concentration of hemoglobin and theformed elements of the blood as a diagnostic test in nutritional studies has beenshown.

Note: We wish to acknowledge our indebtedness to Merck and Co., Rahway, N. J., for some of the crystalline vitamins; to Wilson Laboratories, Chicago, Ill., for the various liver preparations; and to LederleLaboratories, Inc., Pearl River, N. Y., for synthetic folic acid.The authors are grateful to Miss Ethel Thewlis for aid in determining cellular elements and to Drs.Harry A. Waisman and James H. Shaw for assisting in early parts of the work.

     

Hypoplastic anemia due to atabrine

Seven patients with severe hypoplastic anemia were studied at an army TropicalDisease Center. Four of the 7 patients had concomitant dermatitis. The relationship of the prolonged administration of atabrine to the anemia and dermatitis ispresented. A hematologic remission could not be induced by specific therapeuticmeasures. Four of the 7 cases recovered spontaneously.

Note: ACKNOWLEDGMENTThe authors are indebted to Miss Mary L. Boyd for her painstaking supervision of the hematologicstudies.

     

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.

     

Thiamin deficiency in the rhesus monkey; clinical,metabolic and hematologic observations

Seven rhesus monkeys were subjected to one or more episodes of acute thiamindepletion. It is clear that significant metabolic inadequacies preceded demonstrablestructural changes. Diminished food consumption and weight loss were manifestabout two weeks after thiamin was removed from the diet. When the deficiencywas prolonged the animals became apathetic, inactive and progressively weaker.This was followed by ataxia and at times ptosis and tremors. Even in such advancedstates of depletion, administration of thiamin produced dramatic improvement inlocomotion, appetite and reactivity. The blood thiamin content of normal monkeysranged from 5.5 to 10 per 100 ml. of whole blood, values which are comparableto those of healthy human beings. Following withdrawal of thiamin the bloodconcentration fell to values of 4 or less. The tissue content of thiamin was correspondingly reduced in depleted animals. The minimum daily requirement forthiamin calculated on the basis of the time required to redeplete a deficient monkeyfollowing a small dose of thiamin was approximately 15 per kilogram bodyweight. Characteristic degenerative changes in the heart muscle and severe retrogressive changes in the nuclear structures of the central nervous system previouslyreported were noted. Based on careful hematologic studies in 4 animals it isconcluded that thiamin is essential for normal erythropoiesis. Acute or chronicdepletion results in anemia due to suppression of red blood cell formation asindicated by severe depression or absence of reticulocytes in the blood.

Note: ACKNOWLEDGMENTWe are indebted to Miss Mariette Quigley for the blood counts, and to Miss Ruth Johnson for assistance in thiamin assays.

     

Blood exchange in replacement transfusions; studies with erythrocytes tagged with radioactive phosphorus

Red cells labeled with radioactive phosphorus have been used by us to measurethe extent and the rate of the replacement of blood in exchange transfusions. Theexperimental results confirmed the theoretic formulations developed in the preceding paper. On the basis of the above data, it seems reasonable to introduce thesecurves and formulae whenever exchange transfusions are to be performed.

Note: ACKNOWLEDGMENTSWe are indebted to Miss G. Ohno and Miss S. Rezek for technical assistance rendered during thisstudy.

     

THE ERYTHROPOIETIC ACTIVITY OF CHOLINE CHLORIDE IN MEGALOBLASTIC ANEMIAS

1. The effect of the administration of choline chloride has been observed in 10cases of megaloblastic anemia of various types.

2. Choline was without effect in a case of untreated Addisonian perniciousanemia which subsequently responded to parenteral liver therapy.

3. Choline was also without effect in a case of nutritional megaloblastic anemia,in a case of megaloblastic anemia of pregnancy, and in two cases of megaloblasticanemia associated with the sprue syndrome. All these cases had proved refractoryto injections of potent liver extract before the choline was given, and all respondedto subsequent oral liver or folic acid therapy.

4. A significant erythropoietic response to choline occurred in two cases resembling Addisonian pernicious anemia which were refractory to parenteral liverextracts.

Secondary responses followed the administration of choline in two other casesof Addisonian pernicious anemia and in a case of megaloblastic anemia of pregnancy, all of which had already responded to injections of liver extract.

5. The significance of these observations is discussed. It is concluded that choline possesses no direct erythropoietic activity, but that under certain circumstances it may potentiate the effect of liver extracts.

It is suggested that refractory megaloblastic anemias may be divided into twogroups. In one, represented by well known syndromes associated with defectiveabsorption or pregnancy, the lack of response to parenteral liver extracts is notcorrected by choline. In the other, represented by two cases simulating Addisonianpernicious anemia, choline is effective in overcoming, partially or completely, therefractoriness to parenteral liver therapy. Consideration is given to the view thatthe refractoriness of this group results from hepatic dysfunction.

6. The most satisfactory method of administering choline probably consists ofintravenous injections in daily doses of 1 gram. Larger doses given intravenouslyare frequently accompanied by unpleasant side effects, while oral administrationappears to be relatively less effective.

7. It seems unlikely that choline will be of practical value in the treatment ofrefractory megaloblastic anemias, for which oral liver preparations provide themost certain and effective treatment. It is possible, however, that choline may beof use in cases complicated by severe hepatic disease.

Note: Acknowledgment: We wish to thank Dr. L. D. W. Scott for permission to include his patient (case10) in our series.

     

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.

     

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.

     

A method for obtaining living leukocytes from human peripheral blood by acceleration of erythrocyte sedimentation

A simple method for obtaining living leukocytes from human peripheral blood ispresented. It consists of the addition of fibrinogen to heparinized blood. Essentiallyall the leukocytes remain in plasma suspension and essentially all the erythrocytesare sedimented out at the end of one hour.

Note: ACKNOWLEDGMENTThe authors wish to express their thanks to Miss Alma Manieri, who did all the blood counts forthis study, and to Mr. Antol Herskovitz, who prepared the photomicrographs.

     

The effect of radiation on hemopoiesis; is there an indirect effect?

1. The general aim of the investigations here reported has been to obtain evidence for or against "indirect" radiation effects.

2. To this end, twenty-six successful cross circulation experiments (carotid tocarotid anastomoses) have been performed between normal cats and radiated cats.

3. Cross circulation was established in most instances at some specified timeinterval after the radiation of one partner. All intervals up to eighty-two hoursafter radiation of one partner were covered.

4. In seven experiments cross circulation was established and then one animalradiated while the other was shielded. These were considered the most criticalexperiments of the group.

5. Detailed data on leukocyte and lymphocyte counts in the normal animals obtained during an approximately twenty-eight day period of follow-up are presented.

6. These data are not considered to support the thesis of indirect effects peculiarto radiation. A trend toward slightly lowered absolute lymphocyte counts innormal animals after cross circulation was not considered significant, and in noinstance did leukopenia develop in the normal animal.

7. The literature is reviewed and discussed.

Note: ACKNOWLEDGEMENTSThe authors gratefully acknowledge the assistance of Mr. Morey J. Wantman and the Division ofStatistics for the statistical analyses and the valuable assistance of Miss Wilma Kujowski in translatingthe foreign literature.

     

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.

     

弓形虫主要表膜抗原SAG2 DNA疫苗的免疫保护作用研究

目的观察弓形虫pVAX1-SAG2真核重组质粒的免疫保护作用,为弓形虫病的免疫预防提供理论及实验依据。方法大量制备重组质粒,免疫小鼠,3周后同量加强免疫1次,5周后无菌取小鼠脾脏,制备脾细胞,用MTT法测定淋巴细胞转化率;用免疫荧光法测定CD4+、CD8+细胞。用弓形虫RH株速殖子经皮下注射攻击感染,每只鼠接种0.1 ml(约含1000个虫体),观察小鼠存活情况。结果对小鼠的脾脏T淋巴细胞亚群CD4+、CD8+进行分析,与对照组比较,免疫组小鼠CD4+细胞显著增多(P<0.05);而CD8+细胞数各组之间差异无显著性(P>0.05);淋巴细胞转化率各组间差异无显著性(P>0.05);攻击感染后,pVAX1-SAG2免疫组小鼠平均存活(7.8±1.8)d,与对照组比较显著延长(P<0.05)。结论弓形虫pVAX1-SAG2真核重组质粒能在组织内表达,并诱导小鼠产生细胞免疫反应。     

THE ACTION OF PTEROYL GLUTAMIC ACID AND NATURAL SOURCES OF FOLIC ACID ON BLOOD DYSCRASIAS INDUCED BY SULFONAMIDE DRUGS

Sulfanilamide, sulfathiazole, and sulfadiazine have been fed at 1 per cent levelsin highly purified diets and their effect on growth, mortality, and blood dyscrasiascompared with that of sulfasuxidine.

The soluble drugs produce conditions which are similar to those produced bysulfasuxidine. The growth depression is alleviated in large measure in the case ofsulfanilamide and to a lesser extent for sulfathiazole and sulfadiazine by folic acid.liver extract powder, and dried yeast extract as well as by para-aminobenzoic acid,

The blood dyscrasias due to sulfanilamide, sulfathiazole, and sulfadiazine aresevere leukopenia, granulocytopenia, and mild-to-severe anemia. These are uniformly prevented or the severity greatly curtailed by feeding folic acid, liver extract powder, or dried yeast extract. PABA has a lesser effect in the amounts fed.

Liver extract powder seems to have a beneficial effect on growth and mortalitywhich is not shown by the other supplements. Both free and conjugated folic acid(as yeast extract and in liver extract powder 1:20) are active in combating thedyscrasias.

Evidence from in vitro experiments with Str. haemolyticus (B Lancefield) indicates that neither folic acid, liver extract powder, nor dried yeast extract in ratiosto sulfonamide which are effective in preventing the blood dyscrasias will inhibitor block the bacteriostatic action of the sulfonamide drugs in vitro.

It is suggested that the action of folic acid, liver powder, and yeast extract is notwholly explained by alleviating a folic acid deficiency caused by intestinal bacteriostasis due to the drugs, but by an increased demand of the animals for folicacid in the presence of certain sulfonamides.

Note: We are indebted to Harold Buskirk, Alice Bergdahl, Hallie Ferguson, E. M. Stapert, F. La Plante,and Evon Saggio for valuable assistance in carrying out the experimental work reported here.

     

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.

     

The pathogenesis of transferred chloroleukemia in the albino rat

The pathogenesis of transferred chloroleukemia in the rat was studied and thefollowing results were derived. This type of leukemia colonized in an orderlymanner, after intraperitoneal inoculation of leukemic cells, first in the retroperitoneal fat, then almost simultaneously in liver, spleen, lungs, and bone marrow.Only after the leucocyte pool was saturated with leukemic cells did theperipheral blood show an increase in both abnormal and normal components.

Submitted on July 1, 1957 Accepted on September 15, 1957     

RUPTURE OF THE SPLEEN IN INFECTIOUS MONONUCLEOSIS: A CLINICOPATHOLOGIC REPORT OF SEVEN CASES

1. Seven cases of ruptured spleen as a complication of infectious mononucleosisare described and reference made to the 3 cases previously recorded in the literature.

2. It proved possible to make an objective histologic diagnosis of infectiousmononucleosis from well prepared sections of the spleens. The diagnosis wasbased on: (a) a blurred architectural pattern due chiefly to large numbers of atypical lymphocytes diffused throughout the pulp and clumped in the blood sinuses;(b) small, poorly defined follicles, usually without germinal centers, in less thanusual numbers per unit area; (c) cellular "infiltrates," composed largely of normaland atypical lymphocytes, in the capsule and trabeculae, in the adventitia ofsmall intratrabecular arteries, and in the subintimal zone of collecting venoussinuses and intratrabecular veins; (d) swelling of the lining or attached cells ofthe blood sinuses.

3. "Infiltration" of the capsule and trabeculae reached considerable proportions, occasionally to the point of complete dissolution of these structures, andserved as a predisposing cause of rupture. The same changes were noted in intactspleens from fatal cases of infectious mononucleosis.

4. The spleen in infectious mononucleosis was 3 to 4 times normal size andruptured during the third or fourth week of the disease.

5. The importance of trivial injury as the exciting cause of so-called "spontaneous" rupture of the spleen has been emphasized. It is recommended thatextreme caution be employed during attempted palpation of the spleen in a suspected case of the disease. When the diagnosis is obvious, splenic palpation maywell be omitted.

     

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.

     

Synergism of ara-C-imuvert combination in aborting the development of transplanted chloroleukemia in the rat

The development of chloroleukemia in the rat after transplantation of C51 chloroma cells can be aborted by treatment with an exogenous source of differentiating activity (DA) or by stimulation of endogenous DA production by the administration of Imuvert, a biologic response modifier. Success or failure of treatment was determined by the ratio of DA to leukemic cell load. Accordingly, in the present study we examined the hypothesis that treatment with cytosine arabinoside (ARA-C) to reduce the leukemic cell load coupled with Imuvert would be more effective than either given alone. Treatment with ARA-C-Imuvert aborted the development of chloroleukemia in 79% of the animals in contrast to 0%, 9% and 45% in animals treated with buffer, ARA-C, or Imuvert respectively. These results suggest that stimulation of endogenous DA in combination with cytoreductive agents offers a potentially curative treatment for myelogenous leukemia.     

The response of eosinophils in the guinea pig to sensitization,anaphylaxis and various drugs

1. The eosinophilic response of the guinea pig sensitized and reinjected with thespecific antigen varies with the nature of the antigen used, but also with theindividual guinea pig in any groupsensitized and reinjected with the same antigen.

2. Certain antihistamine drugs which abolish anaphylactic symptoms, do notabolish the eosinophilic response.

3. The severity of anaphylactic "shock" symptoms has no influence on theeosinophilic response.

4. Histamine phosphate has no effect on the eosinophil count of nonsensitizedguinea pigs protected by benadryl; it causes a distinct eosinophilic response insensitized animals.

5. Heparin—in the dose injected—produced only an insignificant rise in theperipheral eosinophil count of sensitized guinea pigs; adenosine had no effect.

6. Attempts were made to correlate the eosinophilic response in bone marrow,blood and shock tissue of guinea pigs sensitized and reinjected with a specificantigen. The variation within a wide range of the number of eosinophils in thebone marrow of nonsensitized and of sensitized, reinjected guinea pigs is emphasized.A definite correlation seems to exist between the presence of a large number ofeosinophils in blood and lungs; it is shown, however, that this observation permits only limited conclusions.

7. The factors which account for discrepancies in the interpretation of theeosinophilic response, e.g., nature of antigen, route of administration and characteristics of species, are analyzed.

8. The significance of the findings is reviewed in the light of previous work.

Note: ACKNOWLEDGMENTWe wish to express our appreciation to Miss Alice Sprenger for her patient and efficient technicalassistance.