bluuebunny/crossref_metadata_2025 huggingface dataset:
PERNICIOUS ANEMIA AND RELATED ANEMIAS TREATED WITH VITAMIN B12
Authors: JONES EDGAR , DARBY WILLIAM J. , TOTTER JOHN R.
Published: October 2019

Eleven cases treated with vitamin B12 have been presented. Eight patients with pernicious anemia in relapse responded hematologically. Two patients with mild neurologic involvement were relieved by therapy with B12 alone.

Consideration of the quantities of the crystalline vitamin required to promote maximal erythropoiesis in pernicious anemia indicates that less than about 0.75 µg. daily in doses at intervals of several days will not suffice to establish and maintain blood values as high as does adequate treatment with liver extract. Parenteral daily doses of 1.0 µg. promoted good erythropoiesis in one patient, although it appears that the maximum rate of hemopoiesis may require the initial average daily dose of approximately 3.0 µg.

The reticulocyte count is an unreliable quantitative criterion of activity or adequacy of therapy.

It is suggested that hemopoietic factors in addition to PGA and B12 may be required by some patients to obtain maximal erythrocyte levels.

Vitamin B12, as well as PGA, effects a reduction in the fecal urobilinogen output of patients with pernicious anemia. The significance of this finding is discussed.

No change in urinary excretion of pteroylglutamate or of porphyrin was detected in patients treated with vitamin B12.
Production of Erythrogenin by Organ Cultures of Rat Kidney
Authors: Sherwood J. B. , Robinson S. H. , Bassan L. R. , Rosen S. , Gordon A. S.
Published: October 2019

Organ cultures of rat kidney produce a factor capable of stimulating erythropoiesis in exhypoxic polycythemic mice. This material appears to be renal erythropoietic factor or erythrogenin, since biologic activity was observed only after incubation of the culture media with rat serum. Production of this material was similar in cultures prepared from four different anatomic regions of the kidney, suggesting that erythrogenin may be produced diffusely throughout this organ. Production of erythrogenin was usually insignificant during the first 3½ days in culture but became significant in all four anatomic regions during the second 3½ days. Histologic evaluation of the explants revealed primarily autolysis in the early period followed by the development of epithelial elements, many of which comprised renal tubular structures, during the later culture interval. The newly developed cells had a markedly hypertrophic Golgi apparatus, extensive rough-surfaced endoplasmic reticulum, and numerous polyribosomes, findings consistent with active protein synthesis. This organ culture technique provides direct evidence that the kidney produces an erythropoietic factor and represents a promising model for future studies of this problem.
Treatment of Advanced Malignant Lymphomas With Bleomycin
Authors: Rudders Richard A.
Published: October 2019

Bleomycin, a new antitumor agent reported to lack myelotoxicity, was administered to 43 patients with disseminated refractory lymphomas. Either a weekly or daily regimen was used and in order to minimize the possibility of pulmonary toxicity, the total cumulative dose limited to 5 mg/kg. In 37 evaluable patients objective > 50% responses were observed in 18 (49%). Three patients are still in unmaintained complete remission after 6-9 mo and a fourth relapsed after 13 mo. Myelotoxicity was conspicuously uncommon and serious pulmonary toxicity occurred in only one patient. Bleomycin has significant activity in the malignant lymphomas and its unique toxic properties make it ideally suited for combined use with other standard agents or singly in previously treated patients, particularly those with marrow depression.
The Estrous Cycle and the Hemopoietic Stem Cell in the Mouse
Authors: Lord Brian I. , Murphy Martin J.
Published: October 2019

A large variability in the stem cell content of the spleens of female mice by comparison with that of the bone marrow, or with that of the spleen and bone marrow of male mice, is demonstrated. This variability is shown to be associated with the estrous cycle. Stem cells are assayed by the spleen colony-forming test, either exogenously by transplantation to irradiated recipient mice, or endogenously by colony development in sublethally irradiated female mice. It is shown that, for mice in pre-estrus, the spleens contain up to three times as many hemopoietic stem cells (CFU) as during the estrus and post-estrus stages of the cycle. Although the association may be indirect, it is suggested that the changes in splenic stem cell numbers may be related to the hormonal milieu at the time of observation.
Freeze-Etching Studies of Human Platelets
Authors: Hoak John C.
Published: October 2019

The freeze-etching technique was used to study morphologic features of normal human platelets. A well-developed canalicular system was demonstrated within the platelet, with communication to the plasma surface of the platelets and with direct contact of the canaliculi with organelles. Cross-section fractures of granules revealed compartmentalization in some while others appeared homogeneous. Several types of filamentous structures were observed within the matrix of the platelet. Microtubular subfilaments were approximately 80 Å in diameter. Microfilaments were observed in other areas of platelets and had a diameter of 50-70 Å, a size consistent with that described for actinoid filaments. The interior of the platelet membrane contained 85-Å particles, while the membrane surface had a more granular appearance. Earlier morphologic descriptions of platelets prepared by fixation methods have been confirmed and expanded with this technique.
A Light and Electron Microscopic Study of the Lymphomyeloid Complex in Hypervitaminosis A
Authors: Polliack Aaron , Drexler Ruth
Published: October 2019

Prolonged administration of vitamin A to guinea pigs caused lymphomonocytosis and marked changes in organs of the lymphomyeloid complex, with proliferation of atypical mononuclear cells in the spleen, lymph nodes, and liver. In the bone marrow, atypical mononuclear cell proliferation was also encountered and represented 13% of the entire marrow cell population in some animals. Electron microscopic examination of the marrow revealed many lymphocytes and plasma cells with a well-developed rough endoplasmic reticulum (RER), suggesting that these cells are active in protein synthesis and antibody production. The atypical mononuclear cells described appeared to belong to the monocytic series and contained a variety of dense bodies, and a well-developed RER and Golgi apparatus. Vitamin A is mitogenic, labilizes lysosomal and cell membranes, and can act as an adjuvant enhancing immune responses. It has been suggested that vitamin A alters lymphoid cell membranes and enhances the degree of immune response by stimulation of cell division in immunologically competent cells. The lesions described here may well be the morphologic counterpart of an enhanced cellular immune response.
The Effect of Neutropenia on Myeloid Growth and the Stem Cell in an In Vivo Culture System
Authors: Tyler W. S. , Niskanen Eero , Stohlman Frederick , Keane Jane , Edward Donald
Published: October 2019

An in vivo diffusion chamber (DC) culture technique was used to evaluate the role of circulating humoral factors in the control of myelopoiesis. Normal mouse bone marrow cell suspensions sealed in DC’s were implanted intraperitoneally into mice or rats rendered neutropenic by pretreatment with cyclophosphamide, and the growth of cells in the DC’s was evaluated at intervals thereafter. When marrow was cultured in hosts with graded neutropenia, a dose-related augmentation of myelopoiesis was demonstrated. The growth of macrophages and of pluripotent stem cells, measured by the spleen colony-forming technique, was also enhanced by culture in neutropenic hosts. These data provide further evidence that the rate of myelopoiesis is influenced by a circulating humoral factor. They further suggest that a humoral or hormonal factor is important in control of the pluripotent stem cell population.
The Liver as a Source of Extrarenal Erythropoietin Production
Authors: Fried Walter
Published: October 2019

Nephrectomized rats exposed to intense hypoxia produce sufficient erythropoietin (Ep) to detectably increase their plasma Ep titers. Extrarenal Ep production is no longer detectable if nephrectomized rats are also subjected to 80% hepatectomy prior to being made hypoxic at 0.465 atmos and is barely detectable but significantly lower than in nephrectomized rats when exposed to 0.435 atmos. These results suggest that the liver plays an important role in the production of extrarenal erythropoietin.
Incidence of Monoclonal Proteins in a Minnesota Community With a Cluster of Multiple Myeloma
Authors: Kyle Robert A. , Finkelstein Sidney , Elveback Lila R. , Kurland Leonard T.
Published: October 2019

Multiple myeloma was diagnosed in six residents of Thief River Falls, Minn. in 1968 (a rate of 84/100,000). A survey of the normal population for monoclonal serum protein peaks was undertaken in 1969. Serum from 1200 residents of the city, who were 50 yr of age or older, was collected. Fifteen (1.25%) had monoclonal serum protein peaks: IgG in 11, IgA in one, and IgM in three, with kappa in 11 and lambda in four. No clinical, laboratory, or roentgenographic evidence of myeloma or macroglobulinemia was found in any of the patients. The incidence of monoclonal serum protein peaks is similar to that reported from Sweden. It was concluded that there was no significant increase in monoclonal proteins in the population of Thief River Falls. The 1968 socalled epidemic of multiple myeloma in Thief River Falls remains unexplained.
Erythroblastic Differentiation of Stem Cells in Hemopoietic Colonies
Authors: DeGowin Richard L. , Hoak John C. , Miller Suzanne H.
Published: October 2019

An early phase of cellular replication precedes the initiation of erythropoiesis in hemopoietic spleen colonies of mice receiving a supralethal split-dose of irradiation, first with the leg shielded and then followed by leg irradiation 3 hr later. Previous studies indicated that this proliferative phase represents the repopulation of a depleted, endogenous stem cell compartment. To label replicating stem cells, tritiated thymidine (3HTdR) was injected intraperitoneally in mice on days 1-4 after irradiation. Touch preparations and autoradiograms of splenic colony cells were examined with light and electron microscopy from 2 to 4 days after irradiation and 5-8 days later, when erythroblasts and other differentiated cells first appeared. Replicating mononuclear cells that were pulse labeled during the early proliferative phase of stem cell renewal resembled medium- to large-sized leptochromatic lymphocytes. Electron microscopic examination of autoradiograms demonstrated a labeled undifferentiated cell with a thin rim of nuclear heterochromatin. Sequential studies showed that the injection of 3HTdR on days 1-4, when only the mononuclear cells were present, resulted in labeled erythroblasts and other differentiated cells on days 5 and 6. These results confirm the presence of an early proliferative phase in endogenous splenic hemopoietic colonies and strongly suggest that mononuclear cells, replicating during this period of self-renewal of the stem cell compartment, transform to erythroblasts and other cells.
Marrow Transplantation and Iron Therapy in Mouse Hereditary Microcytic Anemia
Authors: Harrison David E.
Published: October 2019

The defective tissues causing the microcytic anemia in mk/mk mice have been studied by transplantation of marrow erythrocyte precursors into lethally irradiated recipients. Microcytic (mk/mk) marrow in irradiated normal mice produced erythrocyte parameters, blood values, spleen/body weight ratios, and reticulocyte percentages characteristic of the microcytic anemia, but splenic iron stores remained high. Normal marrow in irradiated microcytic mice produced erythrocyte parameters intermediate between microcytic and normal and, combined with iron injections, produced normal erythrocyte parameters, blood values, spleen/body weight ratios, and reticulocyte percentages; long-term splenic iron stores remained low. Iron injections did not improve blood values of either irradiated normal mice implanted with microcytic marrow or untreated microcytic mice. Microcytic mice injected with iron recovered more rapidly from repeated severe bleedings than did uninjected microcytic mice, but iron injections made no difference in final blood values reached. Only slightly less 59Fe remained in mk/mk than in normal mice 22 days after injection. These results indicate that not only the erythrocyte precursor cells but also iron supplies to these cells and splenic iron stores are defective in microcytic mice and that iron loss is not responsible for the iron storage defect.
Growth of Human Bone Marrow in Liquid Culture
Authors: Golde David W. , Cline Martin J.
Published: October 2019

A system for the cultivation of normal and malignant human bone marrow cells in liquid medium is described. The apparatus used is an in vitro diffusion chamber in which cells grow in suspension and upon a dialysis membrane. Proliferation and maturation of normal granulocytes and macrophages were sustained in culture for several weeks in the absence of exogenous stimulating substances. Cell replication was documented by a rise in viable counts, 3H-thymidine incorporation, and labeled mitoses. The entire maturational sequence of the granulocyte and macrophage series was identified in culture, and the cells were characterized by light and electron microscopy, cytochemical properties, phagocytic ability, and the presence of surface receptors for IgG. Nucleated red cell precursors were observed in some cultures for up to 11 days. Malignant cells from patients with various hematologic neoplasms were also successfully cultured. With this system human myeloma cells were maintained in culture for up to 3 wk on primary explant, and continued to synthesize immunoglobulin. The in vitro diffusion chamber technique permits the cultivation of normal human bone marrow cells in liquid medium and provides a convenient means for studying normal and neoplastic hematopoietic cell differentiation and function in short-term culture.
The Hemagglutination-inhibition Assay for Thrombopoietin
Authors: McDonald T. P.
Published: October 2019

A sensitive immunoassay procedure for the detection of the thrombopoietic-stimulating factor (TSF) has been developed. This procedure is similar to the hemagglutination-inhibition (HAI) technique that is available for erythropoietin. For the production of TSF, sheep were made thrombocytopenic by the injection of antiplatelet serum. TSF from sheep sera was partially purified by column chromatography and tested for biological activity in thrombocythemic mice. A TSF-rich serum fraction was used to immunize rabbits, and the immune serum was tested for hemagglutinating antibodies. The hemagglutinating antisera were absorbed with normal sheep serum to remove antibodies not specific to TSF. The absorbed antisera containing TSF-specific antibodies were then used to detect TSF in platelet-depleted sheep sera by use of the HAI assay procedure. The data indicate that TSF can be detected and quantified in sheep serum by use of this technique.
Platelet Dysfunction in Glycogen Storage Disease Type I
Authors: Czapek Emily E. , Deykin Daniel , Salzman Edwin W.
Published: October 2019

A hemorrhagic tendency has been observed in patients with glycogen storage disease Type I (GSD-I). We have studied the hemostatic mechanism in six patients with GSD-I who have mild to severe bleeding tendencies. All exhibited abnormalities of platelet function (decreased prothrombin consumption, abnormal aggregation reactions, prolonged bleeding time, and low platelet adhesiveness). The degree of dysfunction correlated with the patients’ general clinical condition: when patients were acidotic and severely lipemic, platelet function was strikingly impaired. Two patients received a course of continuous intravenous alimentation. As their general metabolic state approached normal, abnormal platelet function was corrected. We found normal platelets to contain no glucose-6-phosphatase. There was no consistent increase in platelet glycogen levels in patients with GSD-I. These data indicate that the bleeding disorder in GSD-I is an acquired defect rather than a primary platelet abnormality.
Mechanisms of Immunosuppression: Effects of Cyclophosphamide on Cellular Immunity
Authors: Winkelstein Alan
Published: October 2019

A course of cyclophosphamide administered to guinea pigs after immunization with mycobacteria led to a transient inhibition of cutaneous hypersensitivity to tuberculin protein. This treatment resulted in generalized depletion of lymphoid cells, including lymphopenia, and a substantial reduction in the number of macrophages found in induced peritoneal exudates. Results of in vivo cell transfer studies indicated that cyclophosphamide-treated recipients were rendered transiently unresponsive to purified tuberculin protein (PPD). However, sensitized lymph node cells from drug-treated donors were capable of transferring immunity to normal recipients. In vitro tests of cellular immunity with lymph node cells from drug-treated animals yielded discordant results. The proliferative response to both PHA and PPD was significantly impaired. However, these lymphoid cells functioned normally in assays for migration inhibition factor (MIF). Thus, these data suggest that cyclophosphamide can limit proliferation without impairing intermitotic functions of sensitized lymphocytes, such as the release of MIF. Results of this study suggest that cyclophosphamide does not impair development of a population of specifically sensitized T-type lymphocytes. However, several components of the expression phase are affected; the resultant anergy is probably due to a summation of effects on lymphocytes and macrophages.
Autoimmune Hemolytic Anemia During Pregnancy With Hemolytic Disease in the Newborn
Authors: Baumann Rolf , Rubin Harry
Published: October 2019

A young woman had idiopathic autoimmune hemolytic anemia occurring in the last weeks of pregnancy. The hemolytic process was quite severe in the mother but responded to steroid therapy and transfusions. The baby was born with a mild form of hemolytic disease with elevated indirect bilirubin levels, increased normoblasts and reticulocytes, and a positive direct antiglobulin test. The child did not require any exchange transfusions, and although the hemoglobin fell to a low of 9.7 g/100 ml 1 mo after delivery, it continued to rise to normal levels after that. The child is developing normally. The antibody was an IgG globulin, presumably 7S, and crossed the placenta to react with the fetal red cells. The antibody showed no specificity when tested with a panel of cells, including Rh null cells.
Defective DNA Synthesis in Human Megaloblastic Bone Marrow: Effects of Hydroxy-B12 5'-Deoxyadenosyl-B12 and Methyl-B12
Authors: Van Der Weyden Martin B. , Cooper Mary , Firkin Barry G.
Published: October 2019

In cobalamin deficiency, inadequate DNA-thymine synthesis appears to result from decreased conversion of N5-methyltetrahydrofolic acid to tetrahydrofolic acid (THF). The N5-methyl THF conversion catalyzed by N5-methyl THF-homocysteine methyltransferase requires a cobalamin coenzyme, presumed to be methylcobalamin (methyl-B12). In support of the above, in B12-deficient marrow cultures, methyl-B12 appears to be the most effective cobalamin form to correct defective DNA-thymine synthesis. This was measured by the ability of deoxyuridine to suppress tritiated thymidine incorporation into DNA. While methyl-B12 produced complete correction of defective DNA synthesis, 5'-deoxyadenosyl cobalamin (5'-deoxyadenosyl-B12), cyanocobalamin (cyano-B12), and hydroxycobalamin (hydroxy-B12) effected only partial correction. The methyl-B12-mediated correction was blocked by methotrexate (MTX). The effect of MTX, in turn, was reversed by THF. In folate-deficient marrows, the B12 analogues did not correct defective DNA-thymine synthesis. The differential effects of hydroxy-B12 and methyl-B12 in correcting defective DNA-thymine synthesis in B12-deficient marrows suggest that the complex mechanisms for N5-methyl THF-homocysteine methyltransferase activation in Escherichia coli may not predominate in human hemopoietic tissue. Since methyl-B12 is the main component of plasma cobalamins, the critical determinant for megaloblastic maturation in B12 deficiency may be the delivery rate of methyl-B12 to marrow cells and its direct activation of N5-methyl THF-homocysteine methyltransferase.
Changes in Globin Synthesis With Erythroid Cell Maturation in Sickle Thalassemia
Authors: Bank A. , Dow L. W. , Farace M. G. , ÓDonnell J. V. , Ford S. , Natta C.
Published: October 2019

Biochemical studies of different groups of patients with sickle β-thalassemia (S-thal) are described. In a group of relatively asymptomatic black patients with sickle thalassemia, there is mild anemia and no clinical symptomatology. α and β-type (βA,βS, and γ) globin chain synthesis in these patients is balanced in bone marrow cells, although there is an excess of α-over β-type chain synthesis in peripheral blood. These mildly affected patients are in contrast to two other groups of S-thal patients of both Mediterranean and black extraction who have anemia and clinical symptomatology and in whom there is absent or more decreased βA-chain production.
Reaction of Normal Human Lymphocytes and Chronic Lymphocytic Leukemia Cells With an Antithymocyte Antiserum
Authors: Aisenberg Alan C. , Bloch Kurt J. , Long John C. , Colvin Robert B.
Published: October 2019

An antiserum against human fetal thymus was prepared in rabbits and absorbed with liver cells from the thymus donor and lymphocytes from a patient with chronic lymphocytic leukemia (CLL). The antiserum reacted with 80%-95% of the peripheral small lymphocytes from 12 normal controls but with only 1%-11% of cells from 12 patients with CLL. The antithymocyte antiserum demonstrates a cell surface antigen present on the majority of normal human lymphocytes, but absent from the neoplastic CLL cell. This finding is consistent with the view that the CLL lymphocyte is devoid of T-cell antigen and with other evidence that this lymphocyte closely resembles the B-cell of experimental animals. It appears likely that leukemic cells will prove useful in preparing an antihuman thymocyte antiserum that reacts exclusively with $$Word$$ cells by removing B-cell reactivity.
An Unusual Cytoplasmic Ribosomal Structure in Pathologic Lymphocytes
Authors: Anday George J. , Goodman Joseph R. , Tishkoff Garson H.
Published: October 2019

Blood slides of a 70-yr-old woman with lymphosarcoma cell leukemia showed 10%-40% notched-split nucleated lymphocytes. A few round, nucleated lymphocytes contained a round or oval body close to the nucleus. The electron microscopic counterpart of this body was a complex tubular structure described previously by D. Zucker-Franklin. The walls of this structure are composed of granules on concentric layers of membranes around a central "core." The ribosomal nature of this structure is evidenced by: (1) similarity of the particles ot ribosomes, (2) the structure’s connections with endoplasmic reticulum, and (3) the ribonuclease effect on the corresponding paranuclear bodies in the blood slides. As an incidental finding, a very few round nucleated lymphocytes showed nuclear blebs only.
The Impaired Responsiveness of Chronic Lymphatic Leukemia Lymphocytes to Allogeneic Lymphocytes
Authors: Smith Martin J. , Browne Elizabeth , Slungaard Arne
Published: October 2019

Normal lymphocytes respond to the presence of allogeneic lymphocytes in culture with a high uptake of 3H-thymidine. We present a study of the interaction of chronic lymphatic leukemia (CLL) lymphocytes with normal allogeneic lymphocytes using the one-way mixed lymphocyte culture technique of Bach. The lymphocytes of 11 patients with CLL were tested separately against those of four normal subjects. In all 11 patients an impaired responsiveness was found. The mean response of CLL lymphocytes to those of the control panel was 19,805 cpm, whereas the mean response of panel lymphocytes to CLL lymphocytes was 90,319 cpm (p < 0.001). There was no significant difference between the mean response of panel lymphocytes to CLL lymphocytes and to those of other panel members (81,630 cpm). Therefore, CLL lymphocytes display an impaired responsiveness to allogeneic lymphocytes, while their capacity to stimulate is intact. These results are in accord with their previously demonstrated impaired reactivity to phytohemagglutinin, a less specific mitogenic stimulus, and serve as further evidence of defective cellular immune response in CLL.
Dihydroxyacetone Metabolism by Human Erythrocytes: Demonstration of Triokinase Activity and Its Characterization
Authors: Beutler Ernest , Guinto Enriqueta
Published: October 2019

Dihydroxyacetone (DHA) was found rapidly to enter human erythrocytes. When incubated with intact red cells, 14C-labeled DHA rapidly appeared in intermediate metabolites and in lactate. Crude hemolysates were found to phosphorylate DHA and glyceraldehyde (GA) but not glycerol. Fifty-fold purification of the phosphorlylating enzyme was achieved by removal of hemoglobin on CM-52 and Sephadex G-200 chromatography. During purification there was no change in the ratio of DHA to GA utilization. The enzyme had a pH optimum of 6,6, a Km of 0.5 µM for DHA, and a Km of 11 µM for GA. With GA as substrate, the Km for (ATP-MG)2- complex was 1.45 mM, and no cooperative interaction was observed. With DHA as a substrate, cooperative interaction with respect to (ATP-Mg)2- complex was found with an n value of 1.4 and a K of 0.44 mM. At temperatures between 15° and 40°C, the enzyme activity gave a linear Arrhenius plot, with deviation from linearity at lower temperature levels. The activity of the enzyme was 0.151 ± 0.011 U/g Hb in five normal subjects, and only slightly increased levels were observed in a patient with marked reticulocytosis. The DHA-phosphorylating activity of erythrocytes is attributed to the activity of triokinase (ATP: D-glyceraldehyde-3-phosphotransferase; E.C.2.7.1. 28). Although DHA is rapidly cleared from blood after injection into rabbits, substantial metabolism is possible for a prolonged time after administration because of the very low Km of the enzyme for DHA. The capacity of red cells to utilize DHA may prove to be useful in the maintenance of 2,3-diphosphoglycerate (2,3-DPG) levels during blood storage and in the in vivo manipulation of red cell 2,3-DPG levels.
IgA-induced Anaphylactic Transfusion Reactions: A Report of Four Cases
Authors: Leikola Juhani , Koistinen Jukka , Lehtinen Marja , Virolainen Martti
Published: October 2019

Anaphylactic reactions after transfusion of a small amount of red cell compatible blood is described. Typical manifestations were substernal pain, dyspnea, and severe normovolemic shock. All the four patients lacked IgA and were found to have antibodies to IgA with titers from 500 to 16,000, as determined by a passive hemagglutination assay. In immunodiffusion against IgA, a single precipitation line was produced by three of the four serums. The antibodies were shown to be IgG by ultracentrifugation, gel filtration, immunoelectrophoresis, and ion exchange chromatography. The antigen-antibody complexes were able to fix complement. Lack of cell-bound immunity was suggested by the failure of IgA to stimulate lymphocytes from one patient. Two of the patients received IgA-deficient blood or plasma; both tolerated the transfusion without ill effects. Twice-washed erythrocytes from normal blood provoked a reaction in the third patient, but after three additional washings, no reaction was observed.
Evaluation of Opsonic and Leukocyte Function With a Spectrophotometric Test in Patients With Infection and With Phagocytic Disorders
Authors: Stossel Thomas P.
Published: October 2019

Paraffin oil droplets containing Oil Red O and coated with Escherichia coli lipopolysaccharide were ingested rapidly by human peripheral blood phagocytes only if they were pretreated with normal serum. This property formed the basis of a screening test in which lipopolysaccharide-coated particles were opsonized with patients’ serums and then were added to autologous leukocytes suspended in two portions, one of which contained nitroblue tetrazolium. After these incubations the cells were washed and extracted with dioxane. Oil Red O and nitroblue tetrazolium formazan in the dioxane extracts were spectrophotometrically assayed, thereby providing simultaneous determinations of the initial rates of ingestion and nitroblue tetrazolium reduction. The test differentiated opsonically-deficient serums and chronic granulomatous disease phagocytes from normals. Serums from individuals with bacterial infections had supernormal opsonic activity, and leukocytes from these patients had increased rates of nitroblue tetrazolium reduction in response to ingestion.
Thrombogenic Activity of Leukocytes
Authors: Niemetz J. , Fani K.
Published: October 2019

Peritoneal leukocytes from rabbits that had been pretreated with endotoxin have a considerable procoagulant activity in vitro. To determine whether these leukocytes were thrombogenic, the leukocytes were infused into normal rabbits. When the leukocytes were infused intravenously, pulmonary emboli and death occurred. When similar numbers of leukocytes derived from control rabbits were infused, no thrombi were found. To investigate the effect of an infusion of leukocytes into the arterial circulation leukocytes were infused into the aorta. When leukocytes from control rabbits were infused, minimal renal cortical necrosis was observed in two out of nine cases; no other thrombi were found, and coagulation changes were minimal. When peritoneal leukocytes derived from endotoxin-primed rabbits were infused into the aorta, renal cortical necrosis occurred in six out of ten cases, and infarction or thrombosis occurred in the lung, liver, spleen, and myocardium. Coagulation test showed changes compatible with the defibrination syndrome, including a fall in the platelet count, fibrinogen level, and levels of factor V and factor VIII, and prolongation of the prothrombin time and partial thromboplastin time. Very high doses of heparin were required to prevent the thrombogenic effect of leukocytes. The relative ineffectiveness of heparin is attributed to an antiheparin effect of the leukocyte. Infusion of rabbit brain thromboplastin extract, which had a tissue factor activity similar to that of infused leukocytes from endotoxin-treated rabbits, caused renal cortical necrosis in 9 out of 15 cases, but the coagulation changes were slight.
Hematopoietic Stem Cell Regulation. I. Acute Effects of Hypoxic-Hypoxia on CFU Kinetics
Authors: Murphy Martin J. , Lord Brian I.
Published: October 2019

This paper, the first of a series on the regulation of the hematopoietic stem cell, examines the effects of stem cell kinetics during an acute exposure to hypoxic-hypoxia. This physiologic perturbation was chosen because of the well-established role it plays in influencing erythropoiesis. BDF1 male mice were exposed to a simulated altitude of 22,000 ft for periods of time ranging from 3 to 48 hr. The following parameters were assessed: Packed red cell volume, number of nucleated cells per femur and spleen, relative and total number of colony-forming units (CFU, i.e., stem cells) in the femoral shaft and whole spleen, as well as the turnover of this population of cells using the "3H-thymidine-killing" technique. The spleen responds by releasing more than one-half (i.e., approximately 108 cells) of its prehypoxic complement of nucleated cells. On the other hand, the femoral cellularity did not decrease but showed a moderate increase. Within 12 hr of hypoxia exposure the total femoral CFU had markedly risen, in some cases between 50% and 75% above control levels. There were, nevertheless, great fluctuations over a short period of time. The population turnover kinetics revealed that after 3 hr of hypoxia an average of 25% of the CFU in both marrow and spleen were in the DNA synthetic phase. It is clear that stem cells are capable of responding rapidly to hypoxia. The possible regulatory mechanisms are discussed in terms of what is known about the kinetics of erythropoietin generation.
A New Human Low Molecular Weight Granulocyte Colony Stimulating Activity
Authors: Price G. B. , McCulloch E. A. , Till J. E.
Published: October 2019

Fractionation of conditioned medium derived from cultures of human peripheral blood leukocytes by ultrafiltration yielded material of low molecular weight, capable of stimulating the formation of granulopoietic colonies by human marrow cells in culture. The molecular weight of the active material was found to be less than that of vitamin B12 (1330 mol wt) on the basis of gel filtration on Sephadex G-25. The low molecular weight colony-stimulating activity (LMW-CSA) was trypsin-sensitive, and extractable into chloroform or diethyl ether. Unlike unfractionated leukocyte-conditioned medium, the LMW-CSA showed no colony-stimulating activity when tested on mouse, rather than human, marrow cell cultures.
Chronic Hemolytic Anemia Due to Cold Agglutinins: A 20-Year History of Benign Gammopathy With Response to Chlorambucil
Authors: Evans Robert S. , Baxter Elizabeth , Gilliland Bruce C.
Published: October 2019

A male, 47, developed chronic hemolytic anemia in association with high titers of cold agglutinins in 1966. His symptoms of the cold agglutinin syndrome began in 1951 with cyanosis of the extremities and face after exposure to cold. In 1956, hemoglobinuria was first noted after chilling. In 1962, the cold agglutinin titer was 1:16,000, and his red cells were coated with complement globulins. In 1966, he developed chronic hemolytic anemia and a macroglobulin M component appeared in the serum protein electrophoresis. The titer of cold agglutinins was 1:64,000. The hemolytic anemia responded to chlorambucil with a decrease in cold agglutinin titer and the level of macroglobulin. Serum complement (C) values returned toward normal. Cessation of chlorambucil was followed by relapse. There was a second response to chlorambucil, but reactivation of the hemolytic anemia occurred following a respiratory infection, despite the reduced titer of cold agglutinin. Chlorambucil was discontinued and has not been administered since. There have been remissions and exacerbations of the hemolytic anemia and a return of the macroglobulin and cold agglutinins toward pretreatment levels. During the past 2 yr there has been spontaneous improvement in his symptoms, and the level of macroglobulin has decreased. Despite the appearance of a monoclonal macroglobulin having kappa light chain and with cold agglutinin activity, there is reason to classify his abnormal immunoglobulin production as benign.
Hemoglobin Function in the Horse: The Role of 2,3-Diphosphoglycerate in Modifying the Oxygen Affinity of Maternal and Fetal Blood
Authors: Bunn H. Franklin , Kitchen Hyram
Published: October 2019

The blood of the newborn horse was found to have a higher affinity for oxygen than that of the mother. This difference was due to the fact that the red cells of newborn foals contained 36% lower 2,3-diphosphoglycerate (2,3-DPG) than red cells from their respective mares. The ATP levels of foal and maternal red cells did not differ significantly. Following birth a prompt rise in the foal's red cell 2,3-DPG occurred, approaching normal (maternal) levels within 5 days. Unlike many other species, the hemoglobins of the newborn and adult horse have been shown to be structurally identical. Furthermore, phosphate-free solutions of newborn and maternal hemoglobins had identical oxygen saturation curves in the absence and presence of added 2,3-DPG. This study demonstrates that, in contrast to other species. the increased oxygen affinity of horse fetal red cells is due to a lower level of the cofactor 2,3-DPG rather than to the presence of fetal hemoglobin.
Serial In Vitro Marrow Culture in Acute Myelocytic Leukemia
Authors: Bull J. M. , Duttera M. J. , Stashick E. D. , Northup J. , Henderson E. , Carbone P. P.
Published: October 2019

The in vitro granulocyte colony-forming ability of bone marrow from 19 patients with acute myelocytic leukemia (AML) was studied at the time of initial diagnosis and serially. Initially 18 of the 19 patients grew scant numbers of colonies, one produced large numbers of colonies. When clinical remission was attained and leukemic blasts in the marrow were reduced to <5%, colony forming ability usually returned to normal. Eight such patients who achieved clinical remission were serially studied during remission. Normal in vitro colony formation ensued in five of the eight patients studied. Three patients did not produce normal numbers of colonies, and it was noted that these patients relapsed within 2 mo. The repeated observation of normal numbers of granulocyte colonies appears to distinguish patients whose remissions are stable from those patients who relapse quickly.
Unbalanced Globin Chain Synthesis in Congenital Dyserythropoietic Anemia
Authors: Hruby Marilyn A. , Mason R. George , Honig George R.
Published: October 2019

Hematologic evaluation of a 5-yr-old girl with lifelong anemia demonstrated the characteristic findings of congenital dyserythropoietic anemia (CDA) type II. Globin chain synthesis was studied in vitro by measuring the incorporation of L-leucine-14C into globin by peripheral blood and bone marrow erythroid cells. In cells from the child and from both of her parents an abnormal balance between the synthesis of the α and non-α globin components of hemoglobin was observed, the α chains being synthesized in excess. Neither parent demonstrated microcytosis, hypochromia, or other findings suggestive of β-thalassemia trait.
Abnormalities of Megakaryocytes in W/Wv Mice
Authors: Ebbe Shirley , Phalen Elizabeth , Stohlman Frederick
Published: October 2019

Megakaryocytopoiesis was evaluated in genetically anemic mice of the W/Wv genotype and was found to be abnormal. Concentration and size of blood platelets were normal. Megakaryocytes were decreased in number in tibial marrow and spleen, and the size of mature megakaryocytes was increased.
Isolation and Properties of Human Platelet Mitochondria
Authors: Fukami Miriam H. , Salganicoff Leon
Published: October 2019

A mitochondrial fraction of human platelets that exhibited respiration coupled to ADP utilization has been isolated. These activities were sensitive to classical inhibitors of mitochondrial respiration and oxidative phosphorylation. Pyridine nucleotide-linked substrates were oxidized at low rates (4-7 natoms/min/ mg), but succinate and α-glycerophosphate were oxidized more rapidly (16 and 38 natoms/min/mg, respectively). Difference spectra showed the presence of the mitochondrial respiratory chain cytochromes, c1 + c, b, and a + a3. Electron micrographs revealed the presence of intact mitochondria with well-preserved outer membranes, as well as various α-granules. There appeared to be no qualitative differences between these human platelet mitochondria and pig platelet mitochondria isolated by the same procedure.
Megakaryocytopoiesis in Experimental Iron Deficiency Anemia
Authors: Choi Sun I , Simone Joseph V. , Jackson C. W.
Published: October 2019

Iron deficiency anemia is associated with increased platelet production. The megakaryocytic response to both moderate and severe iron deficiency anemia has been assessed in rats maintained on a low-iron diet. In animals with moderate anemia, the proportion of immature megakaryocytes and 24-hr tritiated thymidine labeling index were increased with no change in megakaryocyte concentration, size, or mitotic index. This indicates that increased platelet production is achieved by an increased rate of influx of precursor cells into the megakaryocyte compartment with an increased rate of efflux. In severe anemia, however, not only were the labeling index and the proportion of immature cells increased, but megakaryocyte size and mitotic index were significantly increased with a marked decrease in megakaryocyte concentration. This suggests that the circulating platelet mass is maintained by an increased platelet output per megakaryocyte with higher ploidy. Thus, in chronic iron deficiency anemia, megakaryocytopoiesis changes depending on the duration and degree of iron deficiency, with a more rapid turnover at first and increased polyploidy later.
Evidence for the Clonal Origin of Chronic Myeloid Leukemia From a Sex Chromosome Mosaic: Clinical, Cytogenetic, and Marrow Culture Studies
Authors: Moore M. A.S. , Ekert H. , Fitzgerald M. G. , Carmichael A.
Published: October 2019

Cytogenetic studies on a 7½-yr-old child presenting with clinical and hematological features of chronic myeloid leukemia (CML) revealed a constitutional 46 XY/47 XYY mosaicism in skin, blood, and marrow. A third 46 XY, Ph1 cell line predominated in the marrow on initial presentation and in subsequent acute transformation. Assessment of granulocytic colony-forming capacity in agar culture revealed that colony-forming cells (CFCs) were greatly increased in the circulation and possessed the abnormal light buoyant density and low susceptibility to tritiated thymidine killing which distinguishes leukemic CFCs from normal. During acute transformation colony-forming capacity was lost but small, poorly differentiated cell clusters persisted in culture. Only 46 XY, Ph1 metaphase were obtained following cytogenetic analysis of in vitro colonies and clusters displaying leukemic growth characteristics in agar culture. The coexistence of CML and sex chromosome mosaicism in this patient provides further support for the uniclonal origin of CML and indicates that cytogenetic instability implicated in mosaicism may carry an increased risk of further cytogenetic evolution with emergence of the Ph1 chromosome.
Inhibition of Hemoglobin Synthesis by Cyanate In Vitro
Authors: Alter Blanche P. , Kan Yuet Wai , Nathan David G.
Published: October 2019

Cyanate inhibits sickling and prolongs red cell survival in sickle cell anemia. However, cyanate markedly inhibits hemoglobin synthesis in vitro. Incorporation of radioactive amino acid into hemoglobin by human sickle reticulocytes or bone marrow and by rabbit reticulocytes (whole cell or cell-free lysate) was inhibited by as little as 2 mM cyanate and abolished by 50 mM. Both alpha- and beta-S chains were equally affected. The inhibition was only partially reversible by washing the cells after exposure to cyanate. Transport of radioactive amino acid into the cell was not impaired, and free intracellular amino acid was not carbamylated. Aminoacylation of transfer RNA was not inhibited; the acylated amino acid was not carbamylated. Examination of polysome patterns by sucrose density gradient centrifugation revealed degradation of polysomes to monosomes, suggesting inhibition of initiation of protein synthesis by cyanate. In a cell-free lysate, cyanate prevented hemin stimulation of initiation. We conclude that cyanate profoundly inhibits initiation of hemoglobin synthesis in vitro.
Hemoglobin LeporeBoston in Two Iranian Families
Authors: Rahbar S. , Golban-Moghadam N. , Saoodi H.
Published: October 2019

Hemoglobin LeporeBoston was found in two Iranian families presenting the clinical manifestations of β-thalassemia trait. Electrophoresic examination of the patients’ hemoglobin revealed the presence of an abnormal hemoglobin with Lepore characteristics. Fingerprints and amino acid analysis of selected peptides of this variant suggest that it is similar to LeporeBoston.
Regulatory Mechanism of Glutathione Reductase Activity in Human Red Cells
Authors: Yawata Yoshihito , Tanaka Kouichi R.
Published: October 2019

The mechanism by which glutathione reductase (GR) activity is regulated in relation to flavin metabolism was studied in red cells of normal adults, cord blood, and patients with severe metabolic disorders using spectrophotometric, fluorometric, and radiochemical methods. The increased activity of GR in red cells of adult patients with severe hepatic cirrhosis, chronic uremia, and glucose-6-phosphate dehydrogenase (G-6-PD) deficiency is related to the increased per cent saturation of GR with flavin adenine dinucleotide (FAD), and increased red cell flavin level. The per cent saturation of GR with FAD correlates with (1) the degree of clinical severity in hepatic cirrhosis or chronic uremia, and (2) total flavin level in red cells. Thus, the increased GR activity in these patients may be regulated actively or passively by the increased flavin level in red cells. This suggests that the increased requirement for enhanced activity of the pentose phosphate pathway may affect GR metabolism secondarily, leading to the association of GR with FAD through an increased uptake of riboflavin. In contrast to the results in adult red cells, cord erythrocytes show an unexpected metabolic pattern of GR and flavin metabolism. Although the total GR activity of cord red cells is considerably higher than in adult red cells, cord red cell GR is only partly saturated with FAD even in the presence of a significantly increased flavin level in cord red cells. Thus, cord red cells may have an additional control mechanism of GR activity.
Allogeneic Marrow Grafting for Treatment of Aplastic Anemia
Authors: Storb R. , Thomas E. D. , Buckner C. D. , Clift R. A. , Johnson F. L. , Fefer A. , Glucksberg H. , Giblett E. R. , Lerner K. G. , Neiman P.
Published: October 2019

Twenty-four patients with severe aplastic anemia, 14 due to unknown cause, four associated with hepatitis, four drug or chemical related, one with paroxysmal nocturnal hemoglobinuria, and one possibly associated with a Fanconi syndrome did not show spontaneous recovery after 2-24 mo of conventional therapy. Eleven were infected and nine were refractory to random platelet transfusions at the time of admission. All received marrow grafts from HL-A identical siblings. Eighteen were conditioned for grafting by cyclophosphamide (CY), 50 mg/kg on each of 4 successive days, and 6 by 1000 rad total body irradiation (TBI). All received intermittent methotrexate (MTX) therapy within the first 100 days postgrafting to modify graftversus-host disease (GvHD). One patient died on the day of grafting of congestive heart failure possibly related to CY cardiac toxicity. One died on day 6 with septicemia. One died on day 24 without engraftment. Twenty-one patients showed prompt hemopoietic engraftment indicated by rising blood counts, return of marrow cellularity, and in most instances confirmed by blood genetic markers. Four rejected the graft and died on days 33, 41, 51, and 67. Four died between days 45 and 85 with GvHD. One died of cytomegalovirus infection on day 91. One with chronic active hepatitis died on day 427 of unknown causes. Eleven are alive with grafts and without GvHD more than 141, 144, 163, 186, 189, 255, 344, 472, 641, 746, and 823 days after grafting, and ten have returned to normal activity. These results show that normal stem cells will repopulate the marrow in patients with aplastic anemia and demonstrate that long-term stable chimerism is possible in man. They suggest that marrow grafting in patients with complete marrow failure and an HL-A matched sibling should be undertaken before major infections and refractoriness to blood transfusions complicate their course.