Hemolytic reactions produced in dogs by transfusion of incompatible dog blood and plasma; serologic and hematologic aspects

1. Dogs injected intravenously with dog erythrocytes containing one or moreantigenic factors lacking in their own red cells developed iso-hemagglutinins andhemolysins exhibiting characteristics of immune antibodies.

2. Transfusions of incompatible whole dog blood and plasma were carried outunder controlled conditions. Pretransfusion observations were made and followedby closely spaced post-transfusion measurements of serologic and hematologicalterations.

3. The rate of destruction of incompatible donated corpuscles was determinedby tagging the cells with radioactive iron and also by employing the technique ofdifferential agglutination of erythrocytes. It was thereby shown that all of theincompatible donated cells disappeared from the recipient’s circulation withinthe first thirty to ninety minutes following transfusion. The probable mechanismsand relative importance of intra- and extravascular destruction of erythrocytes arebriefly discussed.

4. Destruction of recipient dogs’ corpuscles by donated immune plasma wasrelatively slow, and spherocytosis and increased osmotic fragility of the recipients’ cells were evident for periods as long as twenty days. These observationsare compared with those made in human beings after transfusions of plasma and ofblood from dangerous universal donors.

5. The titer of complement in the sera of recipient dogs was sharply reducedfor at least five hours after all transfusions of incompatible whole blood, but isoagglutinin titers were less regularly reduced after such transfusions.

6. Other notations of interest included estimates of the concentrations of serumbilirubin, sodium and potassium, determinations of clotting time, prothrombinconcentration, and observations on red cell morphology, intravascular erythrophagocytosis, and shifts in distribution of leukocytes and in the electrophoreticpatterns of plasma.

Note: ACKNOWLEDGMENTSIt is a pleasure to acknowledge the technical assistance of Mrs. Jane Peters, Miss Mary Jane Izzo andMiss Shirley Deshon.

     

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.

     

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 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.

     

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.

     

Separation of Lymphocytes, Polymorphonuclear Leukocytes and Monocytes on Glass Columns, Including Tissue Culture Observations

A procedure was described for the separation of lymphocytes, polymorphonuclear (PMN) leukocytes and monocytes on Garvin’s glass bead columns. Dextran-sedimented leukocyte suspensions were added to columnsand incubated at 37 C. Lymphocytes were eluted with fresh serum. Thecolumns were then washed completely free of erythrocytes, platelets andlymphocytes while PMN leukocytes and monocytes continued to adhere. PMNleukocytes and monocytes were released from the glass with EDTA. Monocytes appeared in the effluents somewhat later than the PMN leukocytes,permitting a separation.

A fresh serum, heat labile, Ca^{+ +}- and Mg^{+ +}-requiring PMN leukocyte and monocyte adherence promoting factor was demonstrated andfound to be essential to the procedure.

The viability of column-separated cells was shown by their non-stainingwith trypan blue, motility, phagocytic ability, oxygen consumption, andsurvival or development in tissue culture.

In tissue culture, macrophages developed only from monocytes, whereasNowell’s blast-like, dividing, phytohemagglutinin cells were produced onlyin cultures containing lymphocytes.

Submitted on October 3, 1963 Accepted on December 12, 1963     

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.

     

Congenital Factor VII Deficiency with Normal Stuart Activity: Clinical, Genetic and Experimental Observations

The case of a four year old white girl with a bleeding tendency characterized by the spontaneous occurrence of petechiae, ecchymoses andhemarthroses is presented. Laboratory studies indicate a deficiency of factorVII, with normal levels of all other clotting components, and a vascular deficiency possibly related to frequent upper respiratory infection.

Serum from the patient and from four close relatives having reduced levelsof factor VII is shown to correct the abnormal thromboplastin generation ofStuart factor-deficient serum. The rate of conversion of prothrombin intothrombin is markedly delayed in the patient’s plasma but essentially normalin that of other family members. The patient’s prothrombin conversion iscorrected by the addition of normal or Stuart-deficient plasma. The partialthromboplastin time of the patient is somewhat prolonged.

It is concluded that factor VII deficiency is inherited in this family as anincompletely recessive autosomal characteristic. The penetrance of the deficiency allele in the heterozygous individual appears to be variable. Analysesof the distributions of clotting factor levels in the family and in samplesfrom the normal population indicate that an individual may vary greatly inhis clotting factor levels, the average variation being nearly as great as thevariation among individuals. The magnitude of the variation among siblingssuggests that the penetrance of the factor VII deficiency allele is affected byextrinsic factors and that the observed population variation does not resultfrom the occurrence of multiple alleles. It is suggested that sex may be afactor determining clotting factor levels.

The bleeding of the patient is controlled by transfusion of plasma and byinjection of a plasma preparation called ACC-76 (Behringwerke, Germany),the latter followed by a transient elevation of blood factor VII.

Submitted on March 11, 1959 Accepted on May 18, 1959     

Starch Block Electrophoresis of Plasma and Serum Clotting Factors. Separation of Activated PTC (PTC')

1. Plasma clotting factors separate into two groups on starch block electrophoresis. The contact activation factors—Hageman factor, PTA, and activatedPTA—remain around the origin, whereas the vitamin K-dependent factors—prothrombin, proconvertin, Stuart factor, and PTC—migrate between the-globulins and albumin. AHG, proaccelerin, and thrombin are not recovered.

2. The electrophoretic pattern of serum differs from that of plasma mainly inthe absence of prothrombin and in the presence of activated PTC (PTC’).

3. The electrophoretic mobility of PTC’ is found to differ from that ofnative PTC. This difference may be exploited to separate PTC’ from its nativeform and from Stuart factor.

Submitted on May 12, 1964 Accepted on July 26, 1964     

The relationship between heparin dosage and clotting time

1. The relationship between clotting time and heparin dosage has been studiedin the dog.

2. On the addition of heparin to blood in vitro, a linear relation is found betweenheparin dosage and the logarithm of the clotting time obtained. The sensitivityof the blood sample to the action of added heparin is influenced both by the individual (coagulability of the blood before withdrawal) and by the technics of withdrawal and of determination of the clotting time. It is indicated that alterationsin the latter may be used to extend the range of measurable hypocoagulability dueto heparin. Incubation of heparin with blood for ten minutes increases its anticoagulant effect.

3. When moderate doses of heparin are injected intravenously, five to fifteenminutes are required for the clotting time to reach a maximum. No evidence of abiphasic response was obtained. The maximum clotting time obtained is greaterthan it is with the same amount of heparin added to the blood in vitro, due to theeffect of incubation of heparin with blood on its anticoagulant activity. The in-terval required for the clotting time to return to normal is quite short, and with agiven dosage is constant with different animals. Factors influencing the relationbetween duration of hypocoagulability and dosage are discussed.

4. A test has been devised to determine the sensitivity of the animal to the anticoagulant action of heparin. The clotting time response to certain concentrationsof heparin added to the blood in vitro is determined. A fixed dose of heparin is theninjected intravenously and the clotting time response is again determined. Theresponse in vitro measures the sensitivity of the clotting system to heparin, whilethe in vivo response, when interpreted in the light of the in vitro response, measures the ability of the body to remove heparin from the circulation.

5. By means of this test, it has been determined that anesthesia with pentobarbital decreased the coagulability of the blood, urethane had no effect on coagulability, while the effect of ether was variable. The injection of india ink and evisceration caused a hypercoagulability, while removal of the kidneys had little effect.

6. When the sensitivity of the blood to the anticoagulant action of heparin wastested during these procedures, pentobarbital and nephrectomy had no effect, ethercaused an increase in sensitivity, urethane a decrease. The injection of india inkand also evisceration markedly decreased the sensitivity of the blood to the anticoagulant action of heparin.

7. Anesthesia with pentobarbital, ether or urethane, the injection of india ink,removal of the kidneys, or removal of the gastrointestinal tract, had no effect onthe duration of heparin action in the body.

Note: ACKNOWLEDGMENTWe are greatly indebted to Professor C. H. Best for his interest and encouragement in these studies,and to Dr. J. Markowitz for performing the experimental surgery for us. The study was supported bya grant from the John and Mary R. Markle Foundation.

     

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.

     

Hemorrhagic diathesis associated with the presence of an anticoagulant in circulating blood; case report and laboratory studies

A new observation of a hemorrhagic diathesis associated with the presence ofan anticoagulant in the circulating blood is reported here. The patient was a 21year old male, appearing by clinical evaluation to have hemophilia, but withouta family history of hemophilia. The blood and plasma were strongly anticlottingand had a very long clotting time. The clotting time of recalcified citrated plasmawas greatly delayed by removing the platelets. Freezing and thawing of platelet-rich plasma resulted in a marked shortening of the clotting time. Dilution of theplasma shortened the clotting time, while the addition of calcium and storage ofthe plasma had no effect.

The prolonged clotting time was not corrected by the addition of normal plasmaor plasma fractions having antihemophilic activity. The prothrombin time wasnearly normal. Small quantities of thromboplastin were very effective in shortening the clotting time. The anticoagulant had no antithrombin activity. The "progressive" antithrombin and antifibrinolysin of the patient’s plasma were normal.

The anticoagulant acts during the first phase of coagulation by inhibiting an(plasma) activator of prothrombin. It appears to be identical with the anticoagulant described in three previous publications from the United States.

     

Hypoprothrombinemia; studies of a case of the idiopathic type and the effect of serum administration

1. The reported cases of idiopathic hypoprothrombinemia are reviewed briefly,and a case observed for over three years is presented. Particular attention is calledto the similar clinical pattern presented by the chronic cases.

2. Studies are presented indicating that in this patient the delay in prothrombintime was due, at least in part, to a deficiency of a factor necessary for rapid conversion of prothrombin. This factor, or factors, which we have called Ac-globulin,is contained in a highly active state in fresh normal serum.

3. After the in vitro demonstration of a deficiency of Ac-globulin in the patient’sblood, it was possible to bring about a marked reduction in the patient’s prothrombin time by the intravenous administration of relatively small amounts (15 to45 cc.) of fresh normal (thrombin-free) serum. A further reduction of the prothrombin time to near normal values was brought about by combined wholeblood and serum administration. The evidence suggests that partial correction ofboth prothrombin and Ac-globulin deficiency respectively resulted from suchtherapy.

4. The possible effects of serum and whole blood upon the delayed prothrombinconversion rate of dicoumarolization and liver disease are discussed and preliminary observations in the former type suggest that such therapy may be useful.

Note: ACKNOWLEDGMENTSWe wish to express our appreciation to the following for their cooperation in this study:Dr. McLemore Birdsong, Associate Professor of Pediatrics, University of Virginia Medical School;The Department of Pediatrics, University of Virginia Hospital; The Department of Biochemistry, University of Virginia Medical School; Dr. Myers H. Hicks, Assistant Resident in Medicine, Universityof Virginia Hospital; Dr. Walter H. Seegers, Professor of Physiology, Wayne University College ofMedicine.

     

Liver extract refractory megaloblastic anemia

1 . The patient described in this report had macrocytic anemia, megaloblasticmaturation arrest in the bone marrow, glossitis, hyper-reflexia and diminished vibration perception in the feet. None of these abnormalities was improved by liverextract or vitamin B₁₂ but all responded rapidly to folic acid except the neurologicsigns.

2. This patient appears to have had a megaloblastic anemia which has been described in European clinics under the names "achrestic anemia" and "refractorymegaloblastic anemia." It appears to be similar to "Wills" factor deficiency anemia" and some cases of pernicious anemia of pregnancy.

3. This patient did not appear to have a primary deficiency of folic acid since theexcretion of this substance in the urine was within normal limits. A deficiency ofan unknown factor probably equivalent to "the Wills’ factor" is suggested.

4. It seems likely that folic acid induced a remission in this case by a "massaction" effect. The possible relationship of folic acid, vitamin B₁₂, the unknownfactor and liver extract to nucleo-protein synthesis is discussed.

Note: ACKNOWLEDGMENTWe wish to thank Doctor Charles Foertmeyer for referring this patient to us for the clinical study usedin this report.

     

Angiotensin-II receptor blockers and coronary artery disease: 'presumed innocents': reply

‘Is the jury out’ or ‘is the jury in’? The commentary by Volpe et al.¹ does not appear to recognizethat two important issues in cardiology have recently gainednew insight. First, the reduction of myocardial infarction anddeath with ACE-inhibitors in high-risk patients is greater thanthat derived from blood pressure lowering alone. A ‘bloodpressure-independent’ effect of ACE-inhibitors is supportedby two recent meta-analyses that applied meta-regression, astatistical principal that adjusts for blood pressure differenceswithin trials. One of the analysis included 179 122 patientsfrom trials with ACE-inhibitors or calcium channel blockerswith     

The Antigenicity of Normal and Leukemic Human Leukocytes

1. A leukoagglutinin was formed in the serum of a normal human subjectwho received 10 intravenous injections of blood from a single patient withchronic myelogenous leukemia over a period of 20 weeks.

2. Coincident with development of the leukoagglutinin, first detectable oneweek after the fifth injection of leukemic blood, the normal subject experiencedprogressively more severe febrile reactions to the infusions and exhibited acharacteristic pattern of leukocyte response—namely, an immediate transientleukopenia, followed by a leukocytosis which reached its peak around 3 hoursand subsided to normal within 12 hours.

3. During the early part of the investigation immature leukocytes, presumably from the leukemic donor, could be identified in the recipient’s circulationduring the first hour immediately following injection, but none could befound following the tenth infusion of leukemic blood.

4. The leukoagglutinin which appeared in response to the injections of bloodfrom the single leukemic donor was a typical iso-antibody, showing a broadpattern of reactivity against normal leukocytes from 127 of 129 donors, leukemicleukocytes from 5 of 5 patients with chronic myelocytic leukemia and 6 of 6patients with chronic lymphocytic leukemia. No reactivity was observed againstthe recipient’s own leukocytes, and little or no reactivity was demonstrableagainst the immature leukocytes from 3 patients with acute leukemia.

5. Eighteen months after the last injection of leukemic blood, restimulationof a leukocyte iso-agglutinin in the previously immunized recipient was provoked within one week of commencing a series of intravenous infusions ofblood from a single normal donor.

6. The volume of normal leukocytes employed for the restimulation was 1/10to 1/100 the volume of leukemic leukocytes employed for the primary immunization.

7. The concept of antibody excess was demonstrable in the sensitized recipient. No evidence of in vivo absorption of leukoagglutinin activity was observedafter transfusion of 500 ml. of blood from the normal donor. The severity ofthe recipient’s reaction to the transfused blood was clearly related to the doseof donor leukocytes administered, 0.47 billion cells causing no reaction but4.16 billion causing a moderately severe reaction.

8. Fifteen months after completion of the injections of normal blood, reexposure of the normal subject to injections of blood from a second leukemicdonor resulted in prompt restimulation of leukoagglutinin activity in therecipient’s serum.

9. The leukoagglutinin could be completely absorbed in vitro by incubationwith donor leukocytes.

10. The leukoagglutinin was concentrated in the gamma globulin fraction ofthe recipient’s plasma.

11. The recipient exhibited typical symptomatic reactions and transient hematologic changes following the infusions of leukemic blood.

12. It was possible to correlate the severity of the recipient’s clinical reactionsboth with the strength of the recipient’s leukoagglutinin, as well as with thedose of donor leukocytes transfused.

13. Serologic observations, plus the results of fractionated transfusion studies,indicated that the recipient’s transfusion reactions were related to sensitivityto the donor’s buffy coat (Part II), and more specificially to donor leukocytes(Part III), rather than to donor plasma, platelets or erythrocytes.

14. Sustained stimulation of the recipient’s white cell count as a result of theinjections of leukemic blood was not observed.

15. There has thus far been no evidence of transmission of leukemia to therecipient (now 6 years after the first course of injections of leukemic bloodand 2 years since completion of the present study).

Submitted on July 15, 1960 Accepted on November 20, 1960     

Atheromatous plaque location and arterial remodelling.

See doi:10.1016/S1095-668X(02)00426-8for the article to whichthis editorial refers. Atherosclerosis is associated with a number of structural andfunctional changes in the arterial wall. Endothelial dysfunction,oxidised-LDL accumulation, increased concentrations of macrophages,neutrophils and T-cells as well as smooth muscle cell migrationare some of the most relevant changes that take place duringatherogenesis and coronary artery disease progression. It hasbeen shown that the presence of a space-occupying plaque iscommonly associated with the expansion of the arterial wall(‘vessel remodelling’).¹ The mechanisms responsiblefor this phenomenon are speculative. 1. Positive and negative vessel remodelling The majority of coronary artery atheromatous plaques are eccentricand tend to grow towards the adventitia before encroaching uponthe lumen of the artery. This was shown initially by Glagovet al.¹ in a pioneering study, which demonstrated thatleft maincoronary     

The Effect of Ellagic Acid on Coagulation in Vivo

Ellagic acid shortens the silicone and glass clotting times of the blood ofdogs, cats, rabbits and rats. The silicone clotting time is reduced so as tomake it almost equal to the normal glass clotting time for 5 to 30 minutesafter the injection of the agent. There is occasional acceleration of thromboplastin generation in dogs. No other clotting factors were altered significantlyin our experiments.

There were no obvious toxic effects of ellagic acid, and it appears to decrease bleeding in normal animals.

Submitted on February 15, 1965 Accepted on May 8, 1965     

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.

     

Demonstration by the Nadi reaction of cytochrome oxidase activity in cells of the lymphoid and myeloid series obtained from normal individuals and patients suffering from Hodgkin's and other diseases

Using the Nadi reaction, cytochemical studies of the cytochrome oxidaseactivity of cells of lymphoid and myeloid tissue were carried out. Normal cellsand those from patients with Hodgkin’s disease and leukemia were examined.

With the exception of monocytes and macrophages cells from lymph nodes,spleen, bone marrow and peripheral blood show a low level of cytochromeactivity when compared with myocardial, liver and renal tubular epithelium.

Leukemic cells and those from lymph nodes affected by Hodgkin’s diseasecontain about the same degree of cytochrome activity as their normal counterparts, under the conditions of this study.

The cytoplasmic particles stained by the Nadi reaction correspond in size,distribution and number to particles which can be stained supravitally by janusgreen.

Submitted on March 2, 1951 Accepted on April 18, 1951