bluuebunny/crossref_metadata_2025 huggingface dataset:
Serine and Aspartic Acid Metabolism in Leukemic Leukocytes: Correlation to Effectiveness of Therapy
Authors: DIMITROV N. V. , HANSZ J. , TOTH M. A. , BARTOLOTTA B.
Published: October 2019

Leukocytes from patients with acute granulocytic leukemia (AGL), acute monocytic leukemia (AMoL) and blastic crisis of chronic granulocytic leukemia (BC) were incubated with serine-U-C14 or aspartic acid-U-C14. The metabolism was followed by determination of radioactivity of CO2, lipids, proteins, organic acids, and nucleotides. Leukemic leukocytes in AGL, AMoL, and BC exhibit high serine utilization compared to lymphocytic leukemias. Significant changes in the metabolic pattern of the leukocytes from patients with chronic granulocytic leukemia (CGL) preceded the onset of blastic crisis. Serine utilization remained abnormal in two patients with AGL after complete remission was achieved. Low conversion of serine to CO2 in AMoL was associated with achievement of complete remission and long survival time. Aspartic acid was most actively metabolized by leukemic cells in BC. The results of these studies indicate that the metabolic pattern of serine and aspartic acid varies with the type of leukemic cell. There is evidence that mature polymorphonuclear (PMN) leukocytes in the blood of leukemic patients may possess an abnormal metabolism.
Perturbation of Generation Cycle of Human Leukemic Myeloblasts In Vivo by Methotrexate
Authors: ERNST PETER , KILLMANN SVEN-AAGE
Published: October 2019

The cytokinetic effects of Methotrexate (MTX) on leukemic myeloblasts in vivo in man have been studied, utilizing in vitro and in vivo labeling with 3H-TDR, mitotic index, stathmokinetic index after VCR, measurements of single-cell DNA content, and in vitro labeling with 3H-UDR. It is demonstrated that MTX arrests cells in S for a period (about 20 hr) that corresponds to their DNA synthesis time. This S-phase arresting effect of MTX seems restricted to those cells that are in S at the time of MTX exposure. Cells in G1, G2, and M are not directly influenced by MTX. Some implications of these observations for combination drug therapy and for exploiting differences in the duration of cell cycle phases between leukemic blast cells and differentiating hemopoietic cells to obtain preferential killing of leukemic cells are briefly discussed. In one case, erythroblasts hardly incorporated exogenous TDR and UDR, although their mitotic activity was high. After MTX there was a transient but very pronounced increase in TDR and UDR ultilization. It is suggested that these cells may have had a large intracellular pool of thymidine triphosphate that was temporarily reduced by the action of MTX. Whether these erythroblasts were of leukemic origin remains undecided.
Hemoglobin Köln Disease Occurring as a Fresh Mutation: Erythrocyte Metabolism and Survival
Authors: MILLER DENIS R. , WEED ROBERT I. , STAMATOYANNOPOULOS GEORGE , YOSHIDA AKIRA
Published: October 2019

Electrophoretic and fingerprinting studies in a patient with congenital hemolytic anemia revealed the presence of the unstable hemoglobin Köln (β98val→met). Examination of parents and siblings gave normal results. Extensive blood group and isozyme studies were consistent with the thesis that Hb Köln disease in the propositus was the result of a fresh mutation in one of his parent’s gametes. In the propositus, the activities of enzymes of the Embden-Meyerhof and pentose phosphate pathways were increased, but the level of ATP was decreased. Methemoglobin reduction was delayed when the NADPH-dependent system was utilized with added methylene blue and gave a false-positive result in the glucose-6-phosphate dehydrogenase screening test. Methemoglobin reduction in the absence of methylene blue was normal. Increased methemoglobin and Heinz body formation, decreased osmotic fragility, decreased red cell deformability, and a disproportionate potassium loss without sodium gain occurred with metabolic depletion. The rate of decline of glutathione in propositus’ cells paralleled that in normal cells. Autologous survival of Hb Köln cells was decreased but was not compromised further by oxidant drugs. Marked splenic sequestration of Hb Köln erythrocytes was demonstrated, and an excellent response to splenectomy with improved erythrocyte survival was observed. The intracellular precipitation of unstable globin chains, intracellular dehydration, and increased membrane rigidity probably all contribute to the splenic entrapment of these erythrocytes.
Methemoglobinemia, Heinz Bodies, and Acute Massive Intravascular Hemolysis in Lysol Poisoning
Authors: CHAN T. K. , MAK L. W. , NG RONALD P.
Published: October 2019

Two patients with poisoning by concentrated lysol are reported. One developed methemoglobinemia, a marked decrease in red cell glutathione level, and large solitary Heinz bodies associated with massive intravascular hemolysis 3 days later. The other patient, who had absorbed less lysol, developed methemoglobinemia and multiple small Heinz bodies that subsequently disappeared without frank hemolysis. No preexisting red cell defect was demonstrated in either patient, and in vitro experiments showed that these effects were due to a direct oxidant action of lysol and hydroquinone, a metabolite of phenol on the red cell.
Immune Lysis of Normal and AET-treated Lymphocytes: A Study by Transmission and Scanning Electron Microscopy
Authors: LAMBERTENGHI-DELILIERS GIORGIO , FERRONE SOLDANO , RANZI TULLIO , SIRCHIA GIROLAMO
Published: October 2019

A morphologic study of normal and AET-treated lymphocytes exposed in vitro to cytotoxic antibody and complement (C) has been carried out using both transmission and scanning electron microscopy. The abnormalities observed were of the same kind for the two types of cell, but they were more marked in AET-treated lymphocytes than in their untreated counterparts. These results confirm the serologic finding that AET-treated lymphocytes possess a higher than normal susceptibility to the damaging action of antibody and C in the lymphocytotoxicity reaction.
Turnover Rate of Normal Blood Lymphocytes and Exchangeable Pool Size in Man, Calculated From Analysis of Chromosomal Aberrations Sustained During Extracorporeal Irradiation of the Blood
Authors: Field E. O. , Sharpe H. B. A. , Dawson K. B. , Andersen V. , Killmann S. A. , Weeke E.
Published: October 2019

Lymphocyte kinetics have been studied in four patients treated by extracorporeal irradiation of the blood (ECIB) for immunosuppressive purposes prior to renal transplantation. The technique involves the analysis of chromosomal aberrations sustained by lymphocytes during irradiation and demonstrable in cultures stimulated with phytohaemagglutinin (PHA). Lymphocytes containing aberrations may be regarded as labeled, and this form of labeling has the advantage that it does not require the manipulation of lymphocytes outside the body. Preliminary experiments established that about half of lymphocytes irradiated with doses of 300-400 rad fail to undergo mitosis in culture and that the aberration yield per lymphocyte is the same at the end of a course of ECIB as at the beginning. Irradiated lymphocytes were found to have a mean residence time in the blood of less than 2 min, and for the first few hours of irradiation they were replaced by lymphocytes from tissue pools at a rate such that there was no appreciable fall in the peripheral count. These findings imply that a large proportion of the vascular endothelium acts as a filter for damaged lymphocytes and that initially damaged cells were replaced at the rate of one blood pool per hour; this may represent the normal turnover time of blood lymphocytes. The size of the total exchangeable lymphocyte pool was calculated by two methods depending on a series of basic assumptions concerning the kinetics of irradiated cells. According to the most likely estimate, the total pool contains some 30 times as many cells as circulate in the blood. Forty-two per cent of lymphocytes survived an irradiation dose of 300 rad, 10%-19% survived 380 rad.
Familial Immunopathy With Chromosomal and Functional Abnormalities of Lymphocytes
Authors: Jehn Ulrich W. , Schwartz Robert S.
Published: October 2019

Lymphocytes from members of a family with multiple immunologic disorders had abnormal chromosomes and defective in vitro function. The clinical findings ranged from agammaglobulinemia in the propositus to periarteritis nodosa in his grandnephew. The wife of the propositus was clinically healthy, but her lymphocytes were hyporeactive when cultured with nonspecific mitogens. The high incidence of lymphocyte abnormalities in the children may be the result of the mating of two individuals who have abnormal lymphocytes.
Leukapheresis Therapy of Chronic Lymphocytic Leukemia
Authors: Curtis J. E. , Hersh E. M. , Freireich E. J.
Published: October 2019

The effects of repeated leukapheresis with the IBM Blood Cell Separator were assessed in 13 patients with chronic lymphocytic leukemia (CLL). Five patients had from 14 to 25 leukaphereses at intervals ranging from 7 to 33 days. The median total number of lymphocytes removed was 18.0 x 1011 (range 11.9 x 1011-26.6 x 1011). Two of these patients had a decrease in lymphocyte count and in lymphadenopathy. One of the above and eight additional patients had intensive leukapheresis therapy consisting of five procedures per week for 1-4 wk. The median total number of lymphocytes removed from these patients was 11.34 x 1011 (range 3.30 x 1011-47.4 x 1011). Eight of these patients had a decrease in lymphocyte count, five in spleen size, eight in lymph node size, seven in bone marrow infiltration, and one in liver size. Lymphocyte counts before beginning leukapheresis therapy ranged from 20,500 to 684,000 (median 70,900). The median lymphocyte count on completion of treatment was 19,200 (range 6,590-40,000). After leukapheresis therapy, the median doubling time of the lymphocyte count was 71 days (range 29-118 days). In addition to demonstrating a significant proliferative component in CLL, the objective regression of disease in ten of the 13 patients suggests that leukapheresis is a potentially useful treatment for CLL.
Further Evidence for a Deficient Storage Pool of Adenine Nucleotides in Platelets From Some Patients With Thrombocytopathia—"Storage Pool Disease"
Authors: Holmsen Holm , Weiss Harvey J.
Published: October 2019

The metalobism of adenine nucleotides in platelets was studied in one patient with thrombasthenia and in six patients whose bleeding disorder has been attributed to a defect in collagen-induced platelet aggregation associated with impaired release of platelet ADP (thrombocytopathia). In two of these patients no specific abnormality was found that might account for the defect in the release reaction (thrombocytopathia B). In the other four patients (thrombocytopathia A), significantly decreased amounts of platelet ATP and ADP and an increase in the ATP/ADP ratio were obtained. The specific radioactivity of both ATP and, more strikingly, ADP that was found after incubating their platelets with 3H-adenine was significantly greater than normal. This indicated that the patients’ platelets lacked the nonmetabolic pool of adenine nucleotides present in specialized intracellular granules and that are specifically extruded from the platelet during the release reaction. Low platelet serotonin values were found in three of these patients, indicating that their platelets may lack the entire content of substances normally found in these granules. In all four of the patients with thrombocytopathia A, for which the name "storage pool disease" is proposed, platelet adenine uptake was normal, but increased hypoxanthine formation by resting cells was found in the three patients with low serotonin values. The breakdown of the ATP to inosine monophopshate and hypoxanthine during the release reaction was normal in all patients studied. Platelets from the patient with thrombasthenia were normal in all respects studied.
Alteration of the Cell Periphery During Granulocyte Maturation: Relationship to Cell Function
Authors: Lichtman Marshall A. , Weed Robert I.
Published: October 2019

Physiologic characteristics of the surface of human granulocytes may be important determinants of functional capacity. Studies of immature granulocytes and polymorphonuclear neutrophils (PMN’s) from marrow and blood indicated that immature granulocytes are characterized by a high, neuraminidase-susceptible, negative surface charge density; a high degree of cellular rigidity, as measured by an elastimeter with a micropipette orifice size that approximated estimates of average bone marrow basement membrane pore size (3.5µ); low adhesiveness to glass and plastic; low propensity to aggregate; a slow rate of cell spreading, pseudopod extension, and motility; and a very low rate of phagocytosis. At the myelocyte stage, adhesiveness and phagocytosis, although not prominent, became evident. In contrast PMN’s had a lower negative surface charge density, a higher degree of surface deformability (the entire PMN readily deforming to enter a 3.5µ micropipette whereas the immature granulocyte would not so deform), higher adhesiveness (plastic > glass), a higher propensity to aggregate, a higher rate of cell spreading, pseudopod formation and motility, and a higher rate of phagocytosis. The increased deformability and adhesiveness of the PMN as compared to the immature granulocyte may be a reflection of an altered relationship between relaxing and contracting systems at the cell periphery during maturation and/or changes in the sol-gel state of macromolecules at the cell periphery. This concept is supported by the change in surface properties of the PMN toward those of the immature granulocyte at cold temperature (4°C). The increased deformability and reduced surface negative charge of the PMN could facilitate adhesiveness and pseudopod formation and, thereby, increase cell-surface and cell-particle contact with enhancement of motility and phagocytosis. Hence the exit of PMN from bone marrow and circulation and the functional capacity of the PMN may be determined by a process of cytoplasmic maturation during granulopoiesis that adapts the PMN for its essential cell-extracell interactions.
Acceleration of Fibrin Polymerization by Calcium Ions
Authors: Boyer Markley H. , Shainoff John R. , Ratnoff Oscar D.
Published: October 2019

Calcium ions are known to accelerate the formation of a visible clot upon the addition of thrombin to plasma or fibrinogen. The experiments described demonstrate that this effect of calcium ions is not upon the rate of release of fibrinopeptides from fibrinogen, but rather upon the aggregation of soluble fibrin monomers or polymers, independent of the action of fibrin-stabilizing factor.
An Improved Radioisotope Dilution Assay for Serum Vitamin B12 Using Hemoglobin-coated Charcoal
Authors: Liu Yong K. , Sullivan Louis W.
Published: October 2019

A radioisotope dilution assay for vitamin B12, using hemoglobin-coated charcoal, was modified in several respects, the most prominent of which was the use of cyanide extraction of sera. With these modifications, the B12 levels were significantly higher in sera from normal subjects and patients with folate deficiency, whereas sera from patients with pernicious anemia and other B12-deficient states were only minimally affected. The use of cyanide extraction thus resulted in a clearer differentiation of B12-deficient sera from other sera; all 23 patients with pernicious anemia had B12 levels below 140 pg/ml, whereas all of 85 normal and 218 folate-deficient subjects had B12 levels above 156 pg/ml. The microbiologic assay with Euglena gracilis was similarly affected by cyanide extraction of sera. Thus, it appears that the use of cyanide results in more complete extraction of B12 from serum proteins.
Platelet Function and Pyrophosphates in Osteogenesis Imperfecta
Authors: Hathaway William E. , Solomons Clive C. , Ott John E.
Published: October 2019

Platelet function and serum pyrophosphate (PPi) levels were measured in 15 osteogenesis imperfecta (OI) patients and their parents. The bleeding time was abnormal in five of the subjects. Platelet adhesion to glass was occasionally abnormal (9 of 30 subjects). Platelet factor 3 (PF3) release was impaired in all patients except one and in one or both parents. Platelet aggregation to ADP was abnormal in most of the subjects. Aggregation to collagen was also frequently defective. Thrombin-induced platelet aggregation was always normal. Serum PPi levels were elevated in 62.5% of the subjects. The increased level of PPi was occasionally related to the defect in release of PF3; however, addition of PPi to normal platelet-rich plasma (PRP) did not alter the PF3 release or ADP aggregation. Thrombin-clotted PRP contributed to the elevated PPi but no more in the subjects than in the controls. Presumed carriers of the gene for OI may have defective platelet function and elevated serum PPi, even though no obvious signs of the disease are apparent.
The Effect of L-Asparaginase on the Nucleic Acid Metabolism and Cell Cycle of Human Leukemia Cells
Authors: Saunders E. Fred
Published: October 2019

The effect of L-asparaginase on the cell cycle and nucleic acid synthesis of leukemic cells was studied in five children with acute lymphoblastic leukemia. Following an intravenous infusion of the drug, serial marrow samples were obtained for buffy coat volume, mitotic index, and autoradiographic assessment of DNA and RNA synthesis using tritiated thymidine and tritiated uridine, respectively. A rapid decline in buffy coat volume indicated a lytic effect on lymphoblasts. There was a greater kill of proliferative (blasts in the cell cycle) than nonproliferative (G0) leukemic cells. Mitotic indices changed little until 24 hr; in contrast, thymidine labeling indices decreased markedly to less than 50% of control by 6 hr. The changes in labeling indices prior to changes in mitotic indices indicated that L-asparaginase blocked the entrance of cells into the DNA synthesis period of the cell cycle. Cells already in DNA synthesis appeared to continue into mitosis. Uridine labeling indices decreased progressively in all patients. Uridine uptake was inhibited equally in both proliferative and nonproliferative blasts. Therefore, inhibition of RNA synthesis by L-asparaginase was independent of the proliferative activity of the marrow.
Phytohemagglutinin-induced Lymphocyte Transformation: The Relationship to Prognosis of Hodgkin’s Disease
Authors: Corder Michael P. , Young Robert C. , Brown Robert S. , DeVita Vincent T.
Published: October 2019

Forty-three untreated patients with Hodgkin’s disease were evaluated between 1965 and 1967 with PHA-stimulated peripheral leukocyte cultures. No correlation was found between PHA-induced lymphocyte transformation at the time of diagnosis and the clinical course of the patient’s Hodgkin’s disease. There is also no correlation between lymphocyte transformation and the histologic pattern of nodular sclerosis. Although PHA-induced lymphocyte transformation appears to be a general index of immunologic status, it has not proven of value as a prognostic sign in Hodgkin’s disease.
Granulocyte Chemotaxis in the Chediak-Higashi Syndrome of Mink
Authors: Clark Robert A. , Kimball Harry A. , Padgett George A.
Published: October 2019

Studies of granulocyte chemotaxis were performed in mink with the Chediak-Higashi syndrome. In vivo migration of leukocytes to an inflammatory site was reduced in the affected animals. In vitro studies documented a consistent early impairment in the chemotactic response of Chediak-Higashi mink granulocytes. Serum from Chediak-Higashi mink generated normal amounts of chemotactic factors. The cellular defect in leukocyte chemotaxis in mink is comparable to that observed in humans with the Chediak-Higashi syndrome and may contribute to the increased susceptibility to infection in this disease.
Intraerythrocytic Crystalloid Bodies in Cats
Authors: Altman Norman H. , Melby Edward C. , Squire Robert A.
Published: October 2019

Rectangular-shaped crystalloid bodies (C.B.) were demonstrated within the erythrocytes of intact and splenectomized cats and after incubation in hypertonic NaCl. The morphologic appearance of the C.B. is strikingly similar to that of crystals present in the erythrocytes of individuals with hemoglobin C.
PYRIDOXINE DEFICIENT DIET AND DESOXYPYRIDOXINE IN THE THERAPY OF LYMPHOSARCOMA AND ACUTE LEUKEMIA IN MAN
Authors: GELLHORN ALFRED , JONES LOGAN O.
Published: October 2019

Three patients with disseminated lymphosarcoma and 3 cases of acute leukemia were placed on a pyridoxine deficient diet together with desoxypyridoxine for periods of four to fourteen days. Although there was evidence of malnutrition in the form of weight loss and weakness, none of the signs of specific pyridoxine deficiency described in animals was observed in the human. There was no unequivocal evidence of depression of hemopoiesis, no significant atrophy of lymphoid tissue, no signs of demyelinization of nerves, and no abnormality of tryptophane metabolism determined by urinary xanthurenic acid excretion. The possibility that the dietary restriction of pyridoxine was too brief for the development of a deficiency syndrome was considered, but it was pointed out that the rigors of the procedure were too great to justify more protracted periods of observation.

Two patients had acute toxic manifestations following the administration of large doses of desoxypyridoxine. These were characterized by transient epileptiform convulsions. There were no sequelae and no recurrence of the symptoms when the dose of the drug was reduced.

There was no evidence that the restriction of pyridoxine in the diet together with desoxypyridoxine for periods up to two weeks had any therapeutic effect in lymphosarcoma or acute leukemia. Also, no potentiation of the cytotoxic effect of nitrogen mustard was observed in the patients with lymphosarcoma when chemotherapy was given after the completion of the pyridoxine deficient regimen. It is to be emphasized that this does not absolutely exclude the possibility that pyridoxine deficiency may adversely affect lymphosarcoma in man. The short duration of the experiment and the well known refractoriness to any form of therapy of the tumors in these patients are factors which may have militated against a satisfactory outcome of the regimen.
THE ABSENCE OF THE HETEROPHILE REACTION IN THE SPINAL FLUID IN INFECTIOUS MONONUCLEOSIS
Authors: LYONS HAROLD A. , HARRISON J. GRANT
Published: October 2019

The spinal fluid was studied in twenty cases of infectious mononucleosis proved by clinical picture, blood studies and serological examination. It may be concluded that in acute cases of infectious mononucleosis, the heterophile antibody does not pass into the spinal fluid, but that there may be an increase in cell count, particularly lymphocytes, and in protein content, which is not necessarily proportional to the cell count elevation. These findings have no correlation with central nervous system signs or symptoms.
THE EXPERIMENTAL PRODUCTION OF MACROCYTIC ANEMIA BY OPERATIONS ON THE INTESTINAL TRACT
Authors: CAMERON D. G. , WATSON G. M. , WITTS L. J.
Published: October 2019

1. The literature concerning attempts to produce macrocytic anemia of the liver-deficiency type in animals by operations on the gastrointestinal tract has been reviewed. Operations on the stomach have failed consistently to produce such an anemia, but success has been achieved by operations on the small intestine with the creation of blind loops or intestinal stenosis.

2. The technic we have used to produce macrocytic anemia in the rat is described in detail. The essentials are that the blind loop should fill with peristalsis and that it should not be too low down in the small intestine.

3. Anemia does not usually develop until an interval of several weeks or months after the operation. It is then macrocytic in type and acute in course.

4. The anemia is probably dependent on stagnation in the blind loop and a change in the bacterial flora of the small intestine.
THE BLOOD COUNTS OF THE ADULT ALBINO RAT
Authors: CAMERON D. G. , WATSON G. M.
Published: October 2019

The normal values for blood counts in the adult albino rat are presented.
Regeneration of Marrow Tissue in Chronic Iron Deficiency in Postweanling Rats
Authors: Tavassoli Mehdi , Durocher John R. , Crosby William H.
Published: October 2019

The regenerative potential of chronically iron-deficient rat marrow was studied in extramedullary marrow autotransplants and intramedullary cavity after ablation. Subcutaneous implantation of normal marrow results consistently in establishment of a marrow nodule with a mean weight ratio of 30.7% of the implanted tissue. In iron deficiency, the mean weight ratio was 7.1%, and the take was 11%. After ablation of femoral marrow, the regenerative process proceeds distally from the uninjured marrow at the femoral head. The process of regeneration reached the midshaft after 11 days in iron-deficient animals but took only 5 days for normal animals. Iron repletion accelerated the process toward normal. These results are consistent with biochemical data from iron-deficient marrow (low nucleic acid content, decreased incorporation of 3H-thymidine into DNA, and decreased utilization of 59Fe and 14C-glycine for heme synthesis) and suggest that the reported hypercellularity of the marrow in iron deficiency may reflect sequestration of erythroid precursors, rather than the compensatory mechanism of increased cell proliferation.
Peripheral Cytoplasmic Characteristics of Leukocytes in Monocytic Leukemia: Relationship To Clinical Manifestations
Authors: Lichtman Marshall A. , Weed Robert I.
Published: October 2019

The increased prevalence of tissue infiltration in some cases of monocytic leukemia may represent an enhanced ability of monocytes for tissue entry, continued cell proliferation, an increased life span in the tissues, or a combination of these three factors. Studies in this laboratory have shown that the normal myeloblast undergoes a process of cytoplasmic maturation that adapts it for marrow egress, tissue emigration, and particle ingestion. These include a reduction in surface negative charge density, increased ability to adhere to negatively charged surfaces, increased cytoplasmic deformability, more rapid motility, and enhanced phagocytic rate. In a subject with monocytic leukemia in whom tissue infiltration was a striking feature of the clinical disease, blood monocytes were broadly distributed in regard to their peripheral cytoplasmic characteristics. One portion of the cell population had a high surface negative charge density, was weakly adherent, weakly phagocytic, and poorly deformable, akin to blast cells. A second portion of the cell population had lower surface negative charge density, was adhesive, phagocytic, and readily deformable, akin to mature blood phagocytes. In five patients with myeloblastic leukemia in whom tissue infiltration was not prominent and in whom 92% of cells were myeloblasts, the blood leukocytes had a more homogeneous distribution of surface properties, and these properties were characteristic of immaturity. The increased prevalence of tissue infiltration in some cases of monocytic leukemia may be due to the presence of cytoplasmic maturation, which normally adapts the monocyte for tissue entry.
Bone Marrow Macrophage Precursors. I. Some Functional Characteristics of the Early Cells of the Mouse Macrophage Series
Authors: Cline Martin J. , Sumner Margaret A.
Published: October 2019

Mouse bone marrow cells were cultured in vitro under conditions that allowed little granulopoiesis but permitted proliferation of mononuclear phagocytes. During 10 days’ culture, the population gradually shifted from a preponderance of undifferentiated blast cells and promonocytes to macrophages. Cells up to the A (immature) macrophage stage were capable of DNA synthesis and mitosis. B (mature) macrophages were nondividing. Blast cells were not phagocytic and lacked glass adhesiveness and demonstrable surface receptors for IgG immunoglobulin. With progressive cellular maturation, peroxidase activity disappeared after the monocyte stage. Glass adhesiveness, phagocytic ability, and surface receptors for immunoglobulins appeared to increase with cell maturation from the promonocyte through the macrophage stage of development.
Comparison of the Action of 3H-Thymidine and Hydroxyurea on Testosterone-treated Hemopoietic Stem Cells
Authors: Byron J. W.
Published: October 2019

The treatment of mice with a single, subcutaneous injection of either testosterone propionate or of 5β-dihydrotestosterone results in an increase in the sensitivity of spleen colony-forming cells (CFU-S) to the cytocidal action of high specific activity 3H-TdR. The effect is still present 24 hr after treatment. By 1 hr after exposure in vitro to 2.4 x 10-8M testosterone hydrogen succinate (THS), CFU-S from mouse bone marrow similarly become more sensitive to 3H-TdR. Sensitivity to the cytotoxic action of 10-3M hydroxyurea (HU) also increases. Compared with 3H-TdR sensitivity, however, there is a delay in the onset of sensitivity to HU. The lag is shortened if HU is added 1 hr after the addition of THS. The increased sensitivity of CFU-S to HU is prevented by deoxyribonucleoside, i.e., unlabeled thymidine, and the change during the first hour in 3H-TdR sensitivity is unaltered by the presence of HU. From these data, it is suggested that CFU-S from normal bone marrow contain precursors for DNA synthesis, e.g., deoxyribonucleotides. On the assumption that the action of HU is to inhibit the formation of deoxyribonucleotides from ribonucleotides, the delay in the onset of action of HU may depend on the exhaustion of such pools. It is postulated that CFU-S responding to THS are held up in a stage of the cell cycle close to the G1-S boundary.
Hereditary Defect in Platelet Function in Rats
Authors: Tschopp Thomas B. , Zucker Marjorie B.
Published: October 2019

Hemostatic parameters of fawn-hooded (FH) rats with an inherited hemorrhagic diathesis were compared with those of normal rats. Prothrombin time, partial thromboplastin time, plasma factor VIII, plasma fibrinogen, and platelet count and volume were normal. Bleeding time (BT) in FH rats was over 15 min vs. 1-8 min in controls, and platelet retention in glass-bead columns was reduced. Transfusion of platelet concentrates from normal rats corrected the BT of thrombocytopenic FH rats, but FH rat platelets did not shorten the BT of thrombocytopenic controls. ADP-induced aggregation, measured turbidimetrically in heparinized FH platelet-rich plasma, was normal. Connective tissue did not aggregate platelets of FH rats and released a subnormal amount of 14C-serotonin. In comparison with control platelets washed FH platelets aggregated by thrombin released virtually no ATP and ADP. Platelets, 10,11 of normal rats contained 4.49 µmoles ATP, 0.98 µmoles ADP, and 0.88 µmoles serotonin compared with only about one-half as much ATP and ADP and one-third as much serotonin in platelets of FH rats. These low values suggest the absence of the platelet, "release pool" rather than a defective release mechanism. The hemorrhagic diathesis seen in the FH rats resembles the platelet defect with storage pool deficiency observed in man.2
Morphologic Studies in a Recently Described Dyserythropoietic State
Authors: Freeman Arnold I. , Edwards John A. , Cohen Maimon , Sinks Lucius F.
Published: October 2019

Studies of fetal hemoglobin and bone marrow ultrastructure were performed on a patient with a recently described syndrome consisting of lymphosarcoma, short stature, defective cellular immunity, hypogammaglobulinemia, and mosaicism for a marker chromosome. Abnormalities of the erythroid cells, including multinucleated erythroblasts, nuclear bodies, megaloblasts, and frayed cytoplasm, were seen by light microscopy. Ultrastructure confirmed the findings by light microscopy and showed in greater detail nuclear malformation, loss of nuclear membrane, and loss of cytoplasmic background, as well as peculiar cytoplasmic inclusions. An increased level of fetal hemoglobin (Hb F), ranging between 12%-15%, was detected. The Hb F was discreetly distributed, occurring in a small per cent of the red blood cells. It is speculated that a clone of chromosomally abnormal erythroid precursors might be responsible for producing the erythroblasts with the structural abnormalities described and the erythrocytes with the high hemoglobin F content.
Local Production of Colony-stimulating Factor Within the Bone Marrow: Role of Nonhematopoietic Cells
Authors: Chan Soh Ha , Metcalf Donald
Published: October 2019

Local production of the granulocyte and macrophage regulator, colony-stimulating factor (CSF), within the mouse bone marrow cavity was assessed by determining the capacity of marrow cells or the bone shaft to release or produce active material in liquid cultures. Assays for colony-stimulating activity were performed in agar cultures of mouse bone marrow cells. Following flushing of femoral marrow cells, with or without trypsinization, less than 5% of total marrow cells remained in the bones. However, media conditioned by such bones from adult mice had eight to 20 times the total colony-stimulating activity of media conditioned by the entire extruded marrow cell population. The difference was more marked in femurs from neonatal mice. This activity of both bone and marrow cells was radioresistant. During regeneration following wholebody irradiation, marrow cell activity remained constant, but a significant rise in the capacity of shaft-related cells to conditioned medium occurred simultaneously with the onset of regeneration of granulocytic and monocytic progenitor cells. Local production of CSF by nonhematopoietic cells in the marrow appears to be a significant factor in regulating granulopoiesis and monocyte formation.
Inhibition of the Generalized Shwartzman Reaction by Hypofibrinogenemia
Authors: Bell William R. , Miller Robert E. , Levin Jack
Published: October 2019

The effect of hypofibrinogenemia on production of the generalized Shwartzman reaction (GSR) by the administration of Thorotrast (3 ml/kg) and Escherichia coli endotoxin (50 µg) to rabbits was determined. Severe hypofibrinogenemia (fibrinogen less than 15 mg/ 100 ml) was produced by Arvin, a proteolytic coagulant from the venom of the Malayan pit viper, Agkistrodon rhodostoma. Development of the GSR was inhibited significantly only when hypofibrinogenemia was produced 30 hr before the preparatory injection of Thorotrast and maintained for the duration of the experiment. Severe hypofibrinogenemia of 2-20-hr duration before administration of endotoxin did not block the production of the GSR. Elevated levels of fibrinogen degradation products were demonstrated in the serum of all animals that received Arvin. The concentration of fibrinogen degradation products decreased markedly prior to death only in those animals that developed the GSR. The GSA was not observed in rabbits that received only Thorotrast and Arvin, although these animals had the highest levels of fibrinogen degradation products. The data suggest an essential role for fibrinogen or fibrinogen degradation products and endotoxin in the production of the GSR.
Intravascular Coagulation in Acute Leukemia: Clinical and Subclinical Abnormalities
Authors: Gralnick Harvey R. , Marchesi Sally , Givelber Harry
Published: October 2019

Fifteen patients with acute leukemia were found to have evidence of a generalized hemostatic disorder. These patients could be divided into three groups. The first group consisted of three patients with increased fibrinogen catabolism without clinical or laboratory evidence of intravascular coagulation. The second group of five patients had laboratory evidence of intravascular coagulation without clinically evident bleeding or thrombosis. The third group of seven patients developed symptomatic intravascular coagulation characterized by bleeding, renal failure, and poor response to platelet transfusions. Laboratory evidence for intravascular coagulation in these patients included falling plasma fibrinogen and factor V levels and elevated serum levels of fibrinogen degradation products. Heparin therapy resulted in clinical improvement in all seven patients. Rising plasma fibrinogen and factor V levels correlated with a beneficial clinical response to heparin. Increased fibrinogen catabolism, asymptomatic intravascular coagulation, and symptomatic intravascular coagulation form part of a spectrum of generalized hemostatic disorders in acute leukemia.
Mucopolysaccharide Sulfation in Normal and Leukemic Leukocytes
Authors: Lau Peter , Gottlieb Arlan J. , Williams William J.
Published: October 2019

Mucopolysaccharide sulfation was demonstrated in leukemic granulocytic precursors in short-term suspension culture by incorporation of 35SO4 into a compound identified chromatographically as chondroitin sulfate. The gel filtration pattern of sulfated mucopolysaccharide obtained from leukemic leukocytes was qualitatively similar to that found with normal granulocytic precursors. Sulfation of mucopolysaccharide was about 50% of the normal level in cells from chronic granulocytic leukemia, and approximately 15% of the normal level in cells from acute granulocytic leukemia or chronic granulocytic leukemia in blastic crisis. Lymphocytes from acute and chronic lymphocytic leukemia showed only traces of sulfate incorporation. Leukocyte mucopolysaccharide sulfation was studied in cultured cells from patients with acute leukemia to determine whether this reaction could aid in distinguishing acute granulocytic leukemia from acute lymphocytic leukemia. Significant levels of sulfation were obtained in cells from all leukemias judged to be acute granulocytic, while virtually no incorporation was found in cells from acute lymphocytic leukemia when a 24-hr incubation period was employed. In studies employing a 3-hr incubation, agreement as to the cell of origin of the leukemia and the degree of sulfation in the intracellular fraction was obtained in 16 of 18 determinations performed on 16 patients with acute leukemia. It is proposed that determination of 35SO4 incorporation in the intracellular fraction may help to differentiate acute granulocytic and acute lymphocytic leukemia.
Hypothesis: Engrafted Human Bone Marrow and Blood Cells in Culture
Authors: von Heyden Hans W. , Moore George E.
Published: October 2019

Lymphoblasts appearing in immunosuppressed patients after bone marrow transfusion are compared to those that can be established in vitro as permanent lymphoid cell lines. It is suggested that Epstein-Barr virus (EBV) could be responsible for the recurrent "lymphoblastic leukemia" in these patients and that the transplanted cells may be a clone of nonmalignant cells that has become capable of growing without normal restraints. It is important that in future patients the transplanted cells be characterized as to morphology, chromosome constitution, relative clonability and transplantability, the presence of EBV, T or B cell-like traits, and their growth potential in immunosuppressed heterologous hosts. The antibody titer to EBV should be measured before and after leukocyte transfusion.
Glycogen Metabolism in the Normal Red Blood Cell
Authors: Moses Shimon W. , Bashan Nava , Gutman Alisa
Published: October 2019

Evidence for active glycogen metabolism in normal mature red blood cells (RBC) is presented. Initial rates of 14C-U-glucose incorporation into erythrocyte glycogen were found to be independent of substrate concentration over a range of 3.3-16.6 mM. Incorporation of label into glycogen was initially linear but reached a plateau after a variable period of time that was inversely related to RBC concentration in the medium. The major part of the incorporated radioactivity resided in the outer branches of the glycogen molecule. The optimum pH for 14C-U-glucose incorporation into glycogen was pH 7.6. Replacing the radioactive glucose employed for incorporation after 1 hr of incubation with nonlabeled glucose resulted in a gradual loss of radioactivity from erythrocyte glycogen. In normal cells, glycogen synthesis and breakdown do not result in any significant accumulation of glycogen, whereas in erythrocytes with enzyme defects affecting glycogen breakdown, substantial deposition of glycogen may be observed.
Artificially Induced Thyroid Suppression in Sickle Cell Disease
Authors: Seward Charles W. , Eaton John W. , Chaplin Hugh
Published: October 2019

Depression of thyroid function in patients with hemoglobin SS disease might be expected to: (1) reduce tissue oxygen consumption, (2) decrease erythrocyte 2, 3-diphosphoglycerate, (3) decrease p50, (4) increase the average level of erythrocyte oxygenation, and (5) increase in vivo red cell survival with associated improvement in anemia and possibly in musculoskeletal symptoms. Accordingly, 150-200 mg of 6-n-propylthiouracil were administered three times a day for 143 days to a 21-yr-old male with hemoglobin SS disease. Thyrosuppression was indicated by typical symptoms and appropriate changes in physical and biochemical parameters. Detailed hematologic follow-up, including multiple measurements of red cell mass, 51Cr erythrocyte survival, red cell 2,3-diphosphoglycerate, and p50, showed no change. Furthermore, musculoskeletal symptoms continued in the pattern characteristic of the euthyroid state. A possible explanation for the lack of any change in clinical status may be that decreased cardiac output, with attendant prolonged circulation time and increased tissue oxygen demand per red cell per unit time, offset the absolute decrease in tissue oxygen consumption. The study provided an opportunity to make detailed clinical observations of sickle cell disease in association with thyroid suppression. The results suggest that thyrosuppression within the limits of symptomatic and physiologic tolerance has no therapeutic application in the clinical management of hemoglobin SS disease.
Increase in Circulating Stem Cells in Patients With Myelofibrosis
Authors: Chervenick Paul A.
Published: October 2019

An increased number of granulocytic and mononuclear cell colonies was grown in vitro from blood of patients with myelofibrosis and ranged between 162-4370 colonies/ml of blood. The number of colonies grown from normal individuals and ranged between 40-120/ml blood. There was no correlation between the number of colonies and number of potentially proliferating granulocytic cells (myeloblasts, promyelocytes, myelocytes) plated. In six patients colony size was similar to that of colonies grown from normal individuals and larger than colonies grown from patients with leukemia. Morphologically, colonies consisted of either eosinophils, neutrophils, monocytes, or macrophages. These data indicate that an increased number of stem cells capable of giving rise to in vitro colonies of granulocytes and mononuclear cells circulate in patients with myelofibrosis. The growth pattern of these colonies differed from colonies grown from leukemia cells suggesting that these may not be closely related diseases.
Effect of Testosterone on the Formation of Erythroid Spleen Colonies From Fetal Liver Precursor Cells
Authors: Bleiberg Ilan , Perah Gershon , Feldman Michael
Published: October 2019

Polycythemic x-irradiated female mice, injected with 12-day fetal liver cells, showed 81% suppression of erythroid spleen colonies as compared with nonpolycythemic recipients. On the other hand, in male recipients only 16% suppression was observed. Hence, androgenic hormones seem to play a role in regulating erythropoiesis of explanted fetal stem cells. To test this, we examined the effect of testosterone injected into polycythemic female recipients on the production of erythroid colonies from fetal liver cells. Testosterone was found to alleviate the suppressive effect of polycythemia. Antierythropoietin prevented the appearance of erythroid colonies in testosterone-treated animals. Thus, testosterone seems to act by increasing the levels of, or susceptibility to erythropoietin. Under similar conditions, testosterone did not trigger the formation of erythroid colonies from bone marrow cells in polycythemic recipients. Hence, fetal cells can be induced to form erythroid colonies by doses of erythropoietin that are too low to induce erythroid colonies in bone marrow cells.
Effect of Endotoxin on Granulopoiesis and the In Vitro Colony-forming Cell
Authors: Quesenberry Peter J. , Morley Alec , Miller Marilyn , Rickard Kevin , Howard Donald , Stohlman Frederick
Published: October 2019

The effect of endotoxin on the tibial in vitro colony-forming cell (CFC) and the differentiated granulocyte compartment was evaluated. The injection of 5 µg of Salmonella typhosa endotoxin into CF1 mice leads to elevated serum colony-stimulating factor (CSF) levels along with a transient granulocytopenia followed by the release of granulocytes from the marrow. The present study demonstrates that the number of tibial CFC, as assayed on soft agar, decreases within 20 min after endotoxin, reaches a nadir at 6 hr (65.8 ± 4.9% of control), and then returns to control values by 48 hr. Concomitantly, over the 48 hr after endotoxin, there are sequential increases in the marrow myeloblast-promyelocyte compartment (MPC), myelocyte compartment (MC), and the PMN compartment. Radioautographic studies utilizing 3HTdR showed increased labeling indices and grain counts in the MPC and MC at 24 or 48 hr after endotoxin, indicating a shortening of generation time and perhaps of the DNA synthetic period in these compartments. The present data suggest that elevated CSF levels seen after endotoxin administration may lead to differentiation of the marrow CFC into the granulocyte pathway. There are also changes in the differentiated granulocyte compartment, suggesting a decreased generation time that may be due to CSF or a separate regulator such as antichalone.
Stimulation of Hematopoiesis by Femoral Marrow Curettage in Sublethally Irradiated Mice
Authors: Knospe William H. , Gregory Stephanie Ann , Fried Walter , Trobaugh Frank E.
Published: October 2019

Femoral bone marrow curettage produced an intense stimulus on hematopoietic stem cells (HSC) in sublethally irradiated mice 4 days after curettage. A similar but lesser stimulation of 59Fe incorporation into spleen and red cells was observed. Previous investigations have shown that this effect is not observed in unirradiated mice. Histologic studies of the curetted femurs indicated a rapid regeneration of a primitive mesenchymal tissue within the medullary cavities during the period of stimulation. It is suggested that the primitive mesenchymal cells synthesize a humoral factor that is capable of stimulating HSC in a marrow microenvironment of depleted cellularity. The observations are consistent with a hypothesis of inhibition of HSC response to a similar stimulus in a normocellular marrow by a mechanism of cell-cell interaction. The humoral factor affects HSC similar to fetuin, spleen extract, and alpha globulin fractions.
Platelet Size in Thrombocytopenias and Thrombocytosis of Various Origin
Authors: Kraytman M.
Published: October 2019

Planimetric studies were carried out on canine blood platelets fixed with formaldehyde. In the normal dog, the mean platelet area was 5.25 sqµ (SEM ± 0.34 sqµ). During the first 3 days following acute experimental thrombocytopenia, platelet area increased, averaging 9.5 sqµ (p < 0.01). The reactive thrombocytosis following splenectomy or nephrectomy was not accompanied by a rise in large platelets. Platelet size decreased to 3.9 sqµ in dogs made thrombocytopenic by intravenous injections of mitomycin C. The morphologic changes induced by the various experimental procedures suggest that: (1) newly formed platelets are larger than platelets of a normal population; (2) some large platelets are possibly released by macromegakaryocytes; (3) other large thrombocytes probably represent fragments of granular megakaryocytes; (4) senescence of platelets in the circulation is associated with decreasing size; and (5) large young platelets are preferentially retained in vivo on glass beads.