bluuebunny/crossref_metadata_2025 huggingface dataset:
Enzymic Studies on Phosphatidic Acid Synthesis in Human Platelets
Authors: Okuma Minoru , Yamashita Satoshi , Numa Shosaku
Published: October 2019

Particulate preparations of human platelets were capable of catalyzing acylation of sn-glycerol 3-phosphate by long-chain fatty acyl-CoA thioesters. The principal lipid product formed was identified as phosphatidic acid. The highest specific activity was found in the particulate fraction that sedimented between 12,000 g and 105,000 g. The reaction exhibited a broad pH optimum around 7.4-8.5. The apparent Michaelis constant for sn-glycerol 3-phosphate was 0.48 mM when 28.5 µM palmityl-CoA was used as acyl donor. Palmityl- and oleyl-CoA were better substrates than stearyl-, linoleyl-, and arachidonyl-CoA, as far as the maximal velocity was concerned. Particulate preparations of platelets from normal subjects catalyzed the incorporation of 0.271 ± 0.048 nmole of sn-glycerol 3-phosphate/min per mg of protein. The capacity of human platelets to acylate sn-glycerol 3-phosphate was approximately 40% that of human liver, as compared on the basis of the specific activity of the microsomal fraction. These results suggest that the glycerophosphate pathway makes an essential contribution to the de novo synthesis of phospholipids in human platelets.
Hemodialysis Coil-induced Transient Neutropenia and Overshoot Neutrophilia in Normal Man
Authors: Jensen Douglas P. , Brubaker Leonard H. , Nolph Karl D. , Johnson Charles A. , Nothum Richard J.
Published: October 2019

Profound transient neutropenia, followed by an overshoot of the neutrophil (N) count to an average of 2.5 ± 1.0 (mean ± 1 SD) times higher than average control N count, has been induced in seven normal subjects by reinfusion of heparinized blood that had been stagnated in a hemodialysis coil for 15 min. This was similar to the neutropenia-neutrophilia cycle occurring shortly after the initiation of hemodialysis in uremic patients. At the time of neutropenia, profound monocytopenia was also observed, but only a slight drop in lymphocyte count occurred. Neither monocytes nor lymphocytes subsequently recovered to higher than control values. A DF32P standard N survival procedure, performed in one subject with Hodgkin’s disease in complete unmaintained remission, showed that a large number of unlabeled N appeared during the overshoot, suggesting their marrow origin. Although in two of three attempts neutropenia was produced by reinfusion of blood through the coil without stagnation, no significant overshoot of N count following neutropenia occurred without the stagnation step. Simple phlebotomy and reinfusion of blood without the coil had no effect on the N count. This model may prove useful in studying the possible immediate regulation of the N count by marrow release of N and marrow N reserves in various patients.
Observations on the Settling and Recoverability of Transplanted Hemopoietic Colony-forming Units in the Mouse spleen
Authors: Lord B. I. , Hendry J. H.
Published: October 2019

A series of experiments was designed to study the effect of spleen size on the seeding and retention of spleen colony-forming units (CFU) after injection into irradiated mice. The cellularity of the spleens of recipient mice was varied using a variety of experimental procedures, and the resulting spleen colony production was analyzed with respect to CFU seeding. It was found that the major factor determining the seeding and ultimate retention of CFU in the spleen of a recipient mouse is the size of the spleen. It is concluded that the most reasonable value of the fraction of injected CFU that produces spleen colonies should be determined from the number of CFU recoverable from the fully shrunken spleen. In these mice, the fraction was 9.5%.
Acute Promyelocytic Leukemia: Results of Treatment by Daunorubicin
Authors: Bernard Jean , Weil Marise , Boiron Michel , Jacquillat Claude , Flandrin Georges , Gemon Marie-François
Published: October 2019

Daunorubicin induces complete remissions in about 50% of patients with acute promyelocytic leukemia. The median duration of these remission is 26 mo. Failures are mainly due to hemorrhages as a result of disseminated intravascular coagulation during the first 5 days (25%) or due to sepsis during the second and third week (25%). Long-term survivals are more frequent than in the other acute granulocytic leukemias.
Lymphocyte Lysosomes and Lysosomal Enzymes in Chronic Lymphocytic Leukemia
Authors: Douglas Steven D. , Cohnen Georg , KÖnig Erika , Brittinger GÜnter
Published: October 2019

Electron microscopic cytochemical and biochemical studies of lysosomal markers have been performed in unstimulated normal and chronic lymphotic leukemia (CLL) lymphocytes. Decreased activities of the lysosomal enzymes acid phosphatase and β-glucuronidase but not of the nonlysosomal enzyme malate dehydrogenase were observed in CLL lymphocytes as compared to normal cells. At the electron microscopic level, the number of membrane-bounded acid phosphatase-positive organelles was diminished in CLL cells. (Average 1.07 per cell profile in normal cells and 0.17 in CLL lymphocytes). The findings indicate that the diminution of acid hydrolase activities in CLL lymphocytes is most likely due to a reduced number of lysosomes, rather than to a diminished enzyme content of these organelles.
Disturbed Bone Marrow Cell Proliferation in Primary Shunt Hyperbilirubinemia
Authors: Hamer Jack W. , Fitzgerald Peter H.
Published: October 2019

Cell proliferation studies, utilizing 3H-thymidine radioautography combined with quantitative DNA microdensitometry, demonstrated a gross disturbance of bone marrow cell proliferation in a 17-yr-old youth with primary shunt hyperbilirubinemia. Early erythroid and myeloid cells were arrested at the S, G2 and mitotic phases of the cell cycle, but late normoblasts were not affected. Intramedullary death of the abnormal early erythrmoid cells probably results and is compensated by increased hemopoiesis with a hyperplastic marrow, which 59Fe studies suggest leads to the production and early release of "stress" reticulocytes. The increased early bilirubin production that characterizes this disorder is considered to be derived primarily from intramedullary death of early normoblasts and secondarily from hemolysis in the marrow and spleen of stress reticulocytes.
Recessive Familial Erythrocytosis: Aspects of Marrow Regulation in Two Families
Authors: Adamson John W. , Stamatoyannopoulos George , Kontras Stella , Lascari Andre , Detter James
Published: October 2019

Studies of marrow regulation were carried out in two families with recessively expressed erythrocytosis. The erythrocytosis in the affected individuals was associated with increased erythropoietin (ESF) production. However, hormone production was uninfluenced by alterations in the O2 carrying capacity of the blood when the hematocrit was lowered acutely by phlebotomy. Hemoglobin and red cell function (p50; 2, 3-diphosphoglycerate) in the affected, as well as the unaffected, family members were normal. Renal arteriography revealed no mass lesions and no anatomic abnormalities of the renal macro- or microvasculature. Antisera capable of neutralizing the biological activity of normal human ESF completely inhibited the erythropoietic activity in the serum and urine of those studied. Since increased production of ESF occurred in the absence of recognizable physiologic requirements and since the normal reciprocal relationship between O2 delivery and hormone production was absent, the most likely explanation for this clinical constellation is a cellular defect in the regulation of production of ESF or of its precursor substances.
Immunologic Identification of Tissue Factor (Thromboplastin) Synthesized by Cultured Fibroblasts
Authors: Zacharski Leo R. , Hoyer Leon W. , McIntyre O. Ross
Published: October 2019

Immunologic methods were employed in an attempt to identify a potent procoagulant present in homogenates of human skin fibroblasts cultured in vitro. The activity of this procoagulant was restricted to the early stages of coagulation and was heretofore considered to be due to tissue factor (tissue thromboplastin, factor III) either alone or in combination with one or more of the first-stage coagulation factors (VIII, IX, XI, XII). The present studies demonstrated that procoagulant activity was not diminished by incubation with anti-VIII or anti-IX antibodies of human origin or with anti-VIII antibody of rabbit origin. Furthermore, cell culture homogenates failed to bind antifactor VIII antibody and did not contain an inhibitor of the reaction between factor VIII and its antibody. By contrast, procoagulant activity was obliterated by an antibody to tissue factor protein regardless of whether plasmas deficient in factor VIII, IX, XI, or XII were used in the assay system. The antitissue factor antibody failed to block the procoagulant effect after tissue factor had complexed factor VII. The procoagulant, therefore, consisted entirely of tissue factor.
Cyclic Leukocytosis in Chronic Myelogenous Leukemia: New Perspectives on Pathogenesis and Therapy
Authors: Gatti Richard A. , Robinson William A. , Deinard Amos S. , Nesbit Mark , McCullough Jeffrey J. , Ballow Mark , Good Robert A.
Published: October 2019

Studies were performed on a teen-age female identical twin with cyclic leukocytosis and Ph1-positive chronic myelogenous leukemia (CML), in an attempt to analyze the relationship of these cycles to her disease and to utilize the cycles as reference points for kinetic studies and as landmarks in the progression of her disease. She remained untreated for 2½ yr. Colony-stimulating factor levels showed an inverse relationship to the peripheral leukocyte count. Two distinct phases in cyclic patterns of peripheral leukocyte counts prior to treatment could be appreciated in retrospect: phase 1, a stable 15-mo period during which leukocyte counts peaked every 10 wk and then returned to baseline levels, and phase 2, an 18-mo period of insidious deterioration during which the low points of successive cycles became progressively higher and leukocytes were accumulating in the peripheral blood at a rate of 400 x 106/day. We propose that the increased marrow production of myeloid cells in this disease does not represent a life-threatening situation until cell accumulation begins. We attempted, therefore, to halt or to reverse this accumulation by cell separator leukapheresis.
Effects of Vitamin A on the Erythrocyte Membrane Surface
Authors: Murphy Martin J.
Published: October 2019

The scanning electron microscope was used to examine the effects vitamin A has on the surface of human erythrocytes. The hemolytic properties of vitamin A, both time- and dose-dependent, are confirmed, and the observations are extended to the establishment of a sequence of cellular events that leads to the destruction of the red cell. The biochemical means whereby vitamin A induces these changes are discussed. In addition, the present study indicates that only selective areas of the erythrocyte membranes are responsive to the action of vitamin A, thereby supporting the proposal that the erythrocytic membrane may not be a homogeneous mass.
Radioautographic Observations on Iron Absorption by the Duodenum of Mice With Iron Overload, Iron Deficiency, and X-linked Anemia
Authors: Bédard Yvan C. , Pinkerton Peter H. , Simon Géerard T.
Published: October 2019

The uptake of iron by the absorptive cells of the duodenum, and its subsequent transfer to the lamina propria, has been studied using 55Fe and high-resolution radioautography in mice rendered iron deficient by diet, in mice with dietary iron overload, and in mice with hereditary malabsorption of iron (s/a). In all, as in normally iron replete mice, two phases of iron absorption can be distinguished. There is an early active phase of uptake and transfer, followed by a second "storage" phase. The pathway of iron absorption is qualitatively similar to the normal with rapid uptake by the brush border, passage through the terminal web to the rough endoplasmic reticulum, and areas rich in free ribosomes. Iron passes through the lateral cell membrane, intercellular spaces and epithelial basement membrane to the vessels of the lamina propria. The rough endoplasmic reiculum and areas rich in free ribosomes form the major localization of iron at all stages of absorption. Only a relatively small amount of radioactive iron is found over ferritin in iron-loaded mice and mice with s/a; morphologically recognizable ferritin was not observed at any stage in iron-deficient animals. Significant numbers of grains have not been seen over mitochondria in any group of animals studied. In iron-deficient mice, the rate of uptake and of transfer of iron is increased. With the 10 µg dose of iron used, all iron taken up by the absorptive cell is transferred to the lamina propria within 3 hr, with none remaining in the cell during the second, storage phase. In mice with iron overload, transfer of iron is decreased, resulting in considerable storage of iron in the cell. In s/a mice, uptake does not appear to be unduly reduced, but transfer is reduced, thereby resulting in increased storage of iron in the absorptive cell. The rough endoplasmic reticulum and areas rich in free ribosomes appear to play an important role in the uptake, transfer, and storage of iron by the absorptive cell regardless of the state of the body iron stores. Ferritin would seem to have a less important part in iron absorption, possibly acting as a storage or detoxifying mechanism.
Sickle Cell Safari
Authors: Bemis Edwin L.
Published: October 2019

A narrative concerning mass screening of a large population at risk for sickle hemolobin, glucose-6-phosphate dehydrogenase deficiency, anemia, and lead poisoning is presented. At the time of writing, more than one-tenth of a black community totaling over 104,000 individuals have been tested. The manner of community education and organization, laboratory methodology, results obtained, means of reporting, and follow-up procedures are detailed.
Treatment of Thrombotic Thrombocytopenic Purpura With Antiplatelet Drugs
Authors: Amir Jacob , Krauss Stephen
Published: October 2019

Thrombotic thrombocytopenic purpura (TTP) is a disease with an extremely high mortality rate. Many modalities of therapy have been tried with very limited success. Lately, antiplatelet drugs have been proposed in the treatment of TTP. We report a well-documented case of TTP that presented with severe hemolytic anemia, thrombocytopenia, neurologic manifestations, kidney involvement, and fever. The patient did not respond to splenectomy, corticosteroids, and heparin sodium, but she made a full recovery after antiplatelet drugs (aspirin and dipyridamole) were added to the therapeutic regimen. The coagulation studies showed thrombocytopenia, hypofibrinogenemia, and a positive protamine sulfate test, indicative of disseminated intravascular clotting. The spleen showed numerous arterioles occluded with subendothelial hyaline material. The platelet investigations during antiplatelet therapy revealed impaired platelet aggregation and clot retraction that became normal after discontinuance of the drugs. The patient has now been in full remission more than 12 mo.
Hematopoietic Stem Cell Regulation. II. Chronic Effects of Hypoxic-Hypoxia on CFU Kinetics
Authors: Lord Brian I. , Murphy Martin J.
Published: October 2019

Hypobaric-hypoxia, an established stimulator of erythropoiesis, was used to perturb normal steady-state hematopoiesis and thereby to facilitate an analysis of stem cell kinetics. BDF1 male mice were exposed to a simulated altitude of 22,000 ft for 1-15 days. The following parameters were assessed at daily intervals: packed red cell volumes (PCV), femoral and splenic nucleated cellularity, the relative concentration 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. By 10 days of hypoxia the PCV had risen to 65% and by a fortnight was elevated to, and stabilized at, 70%-75%. After an evanescent increase, the femoral cellularity became slightly hypocellular but returned to normal by 3 days, thence becoming hypercellular from days 5 to 15. The spleen was markedly depleted of cells for 48 hr and recovered to hypercellular levels between 4 and 6 days. It then oscillated from hypercellular to normal levels with a period interval of 5 days. Both femoral and splenic CFU demonstrated oscillations, as did the estimations made on the state of CFU turnover. The femoral and splenic CFU did not decrease in number during the 15 days of hypoxia, indeed, they both revealed cyclic increases. These results are discussed in terms of CFU kinetics and humoral mediation.
In Vivo Aging of Transfused Erythrocytes and 2,3-Diphosphoglycerate Levels
Authors: Dickerman Joseph D. , Ostrea Enrique M. , Zinkham William H.
Published: October 2019

Levels of 2,3-diphosphoglycerate (2,3-DPG) and oxygen affinity were measured in a patient with refractory congenital hypoplastic anemia in order to determine the response of aging transfused erythrocytes to a decrease in tissue oxygen tension. Ferrokinetic studies revealed that the rate of erythropoiesis after transfusion was approximately 10% of normal. In the interval between transfusions, the level of 2,3-DPG and P50 rose to values considerably above the normal range as the level of the hematocrit decreased. On five separate occasions, the 2,3-DPG concentration was elevated (average 25.5 µmoles/g of hemoglobin) and the P50 level increased (average 33.2 mm of mercury) 24 hr prior to transfusion. It is concluded that in vivo aging of transfused erythrocytes does not prevent these cells from responding to an increasingly hypoxic environment by elevating 2,3-DPG levels and by decreasing oxygen affinity.
Identification of Cells in Primate Bone Marrow Resembling the Hemopoietic Stem Cell in the Mouse
Authors: Dicke K. A. , van Noord M. J. , Maat B. , Schaefer U. W. , van Bekkum D. W.
Published: October 2019

The colony-forming unit culture (CFU-C) in the thin-layer agar colony technique is considered to be representative for hemopoietic stem cells (HSC), according to our studies in mouse and monkey bone marrow. Using this in vitro assay as a guide, stem cell concentrates were prepared from monkey and human bone marrow by repeated density gradient centrifugation. The number of CFU-C could be enriched up to 70-100-fold. In such concentrated CFU-C suspensions, a cell, morphologically identical with the hemopoietic stem cell in the mouse (MSCLC, mouse stem cell-like cell) was frequently observed, using a May-Grünwald-Giemsa (MGG) staining method and electron microscope techniques. In MGG-stained preparations, the MSCLC superficially resembles the small lymphocyte; therefore, a staining method has been described, the polychrome procedure, by which both cell populations could be clearly distinguished. Since a fair correlation exists between the number of MSCLC and the number of CFU-C in a variety of primate hemopoietic suspensions, we concluded that the MSCLC might be a good candidate for being the HSC in monkeys and man.
Correlation Between Cytokinetically Resting Lymphocytes and Bone Marrow Restoration: Experiments Using a Discontinuous Albumin Gradient
Authors: Haas Rainer J. , Flad Hans-Dieter , Fliedner Theodore M. , Fache Ingeborg
Published: October 2019

Rats were labeled with 3H-thymidine (3H-TdR) in utero and for 6 wk after birth in order to obtain 100% labeling of all bone marrow cells. Six weeks after the last 3H-TdR injection only cytokinetically resting cells were still labeled. At this time, the regenerative capacity of fractions obtained after centrifugation on a discontinuous albumin gradient was tested in 1200-R x-irradiated recipients, and the results were compared with the effect observed after transplantation of the same number of unfractionated bone marrow cells. One of the fractions obtained had a regeneratory capacity tenfold that of unfractionated cells. In contrast, when the response to a PHA stimulation test was evaluated, this fraction showed a decreased incorporation of 14C-TdR as compared to other fractions. In a second test system, fractions of bone marrow from 3H-TdR-labeled donors made hypoplastic by repeated injections of hydroxyurea were transfused into 1200-R x-irradiated recipients. Regenerative capacity was similar to that seen in the first experiment. The findings indicate a correlation between cytokinetically resting, small mononuclear cells and the regeneratory process in lethally x-irradiated recipients.
Survival in Chronic Lymphocytic Leukemia
Authors: Zippin Calvin , Cutler Sidney J. , Reeves William J. , Lum Diana
Published: October 2019

Survival of 839 chronic lymphocytic leukemia patients has been reviewed in relation to a number of patient and disease characteristics. Over-all, the relative 5-yr survival rate was 44%. Studying survival by age and sex led to the following observations: Dividing age into intervals < 50, 50-59, 60-69, 70-79, and 80 yr and over, relative survival declined with increasing age; relative 5-yr survival for females was higher than that for males-50% versus 41%. This pattern of superior survival of females over males was noted in all but one of the age intervals. Survival was negatively associated with the presence of recorded clinical signs and symptoms, hematological abnormalities, and pathological bone marrow findings. Differences in survival were also found by treatment category and interval from diagnosis to initiation of treatment. Adjustment for differences in distribution of each of these variables did not materially diminish the survival differences noted by age and sex. An unexpected pattern of survival in relation to white blood count level was noted. Survival increased with increasing white blood count at diagnosis, peaking in the interval at 25,000-49,000, and decreasing after that. When survival by white blood count was adjusted for some variables which were found to be associated with survival, the gradient was still noted, though somewhat reduced.
Antibodies to Factor VIII. I. Variations in Stability of Antigen-Antibody Complexes in Hemophilia A
Authors: Allain J.-P. , Frommel D.
Published: October 2019

Human factor VIII-anti-factor VIII complexes were formed in vitro in slight antigen excess, using plasma of hemophiliacs who were found to have antibodies neutralizing AHF activity. These complexes, stable at +37°C and pH 7.4, were submitted to classical procedures known to favor dissociation of antibody from antigen. The methods used to obtain dissociation, incubations at +56°C and at pH 4.2, inactivated both unbound factor VIII and that released as a consequence of dissociation. The extent of dissociation was measured by the recovery of anti-factor VIII activity. Increasing resistance of complexes towards dissociation was observed in the plasma of the patients whose titer of inhibitor was increasing after recent transfusions. These observations suggested the emergence, as a direct consequence of renewed antigenic stimulation, of a population of different antibodies characterized by higher combining strength.
Hemoglobin D Iran α2A β222-Glu→Gln in Association With Thalassemia
Authors: Rohe Ronald A. , Sharma Vijay , Ranney Helen M.
Published: October 2019

A 25-yr-old Indian (Asiatic) woman investigated for a life-long anemia was found to have a hitherto undescribed structural hemoglobin variant α2Aβ222-glu→gln, which was found independently and designated Hb D Iran by Rahbar in members of a family from Iran. In the present case, Hb D Iran was found in association with high A2 thalassemia. The replacement of glutamic acid by glutamine at β 22 (helical residue B4) was demonstrated by thin-layer chromatography after automated Edman sequencing. This is the fourth substitution to be described at β 22; the previous substitutions were (1) G Coushatta (ala); (2) E Saskatoon (lys); (3) G Taipei (gly). Helical residue B4 is an external residue, does not participate in α-β or protein-heme contacts and hemoglobin D Iran resembles Hb A in visible spectra, O2 equilibria in dilute solutions and heat stability. Since the propositus and her mother who was also heterozygous for β thalassemia had similar degrees of anemia, no interaction between this variant and β thalassemia was evident. The presence of this Hb variant in Iranians and in Western Indians may reflect the migrations of populations in these areas centuries ago.
Pseudothrombocytopenia: Manifestation of a New Type of Platelet Agglutinin
Authors: Shreiner David P. , Bell William R.
Published: October 2019

Spuriously low electronic platelet counts were obtained on blood collected in EDTA from six patients. Platelet numbers on blood smears and electronic counts performed on blood collected in other anticoagulants were normal. Plasma or sera from these patients agglutinated homologous, as well as autologous platelets, in the presence of EDTA or DTPA, but agglutination did not occur in blood or platelet-rich plasma anticoagulated with heparin, oxalate, or citrate. Agglutination in the presence of EDTA was not inhibited by heparin, α-tocopherol, or N-ethylmaleimide, but was inhibitied by excess EDTA or divalent cation. The agglutinin was temperature independent in five of the six patients and had the physical and immunologic characteristics of a gamma globulin. The clinical significance and mechanism of EDTA-dependent agglutination of platelets is unknown.
Pancytopenia and Leukemia in Hodgkin’s Disease: Report of Three Cases
Authors: Weiden P. L. , Lerner K. G. , Gerdes A. , Heywood J. D. , Fefer A. , Thomas E. D.
Published: October 2019

Three patients with Hodgkin’s disease who developed severe pancytopenia terminating in acute leukemia (two patients) or preleukemia (one patient) are presented. Prior treatment consisted of radiation therapy alone in two patients, and radiation plus chemotherapy in the third. The only patient in whom chromosomal analysis was performed showed an abnormal chromosome. Therapeutic radiation exposure is thought to contribute to the development of leukemia in such patients. Leukemia is an uncommon complication of Hodgkin’s disease, but is being reported with increasing frequency. The rapid evolution of more aggressive modes of therapy makes it imperative that we seek more information regarding the incidence of this complication in order to achieve maximum therapeutic benefit for patients with Hodgkin’s disease.
Granulopoiesis in Neutropenic Disorders
Authors: Greenberg Peter L. , Schrier Stanley L.
Published: October 2019

Owing to printers’ errors, the following errata appeared in the June 1973 issue of Blood (Vol. 41, No. 6).

"Granulopoiesis in Neutropenic Disorders" by Peter L. Greenberg and Stanley L. Schrier (pp. 753-769).

On page 753, line 3 of text, the footnote for reference I should be deleted and placed after the footnote at the bottom of the page that says "Presented in part at the American Society of Hematology Meeting, San Francisco, Calif., December 7, 1971."

On page 759, in the footnote to Table 4, third line, "22-cell" should read "200-cell."

On page 763, the second sentence of the first complete paragraph should read "CFC values could have been altered by processing causing cell selection."
Serial In Vitro Bone Marrow Culture in Acute Lymphocytic Leukemia
Authors: Duttera M. J. , Bull J. M. , Northup J. D. , Henderson E. S. , Stashick E. D. , Carbone P. P.
Published: October 2019

Serial in vitro bone marrow cultures have been done on eight patients with acute lymphocytic leukemia. Colony-forming activity shows a rapid recovery from low values into the normal range in all patients who achieved stable remissions. Mean values for the stimulated cultures remained stable throughout consolidation, whereas the mean values for the unstimulated cultures declined during the same period. Relapse in two patients was preceded or accompanied by a concomitant decline in colony growth. Colony forming activity appears to be inhibited in a nonlinear fashion by increasing numbers of leukemic cells.
Interacting Cell Populations Affecting Granulopoietic Colony Formation by Normal and Leukemic Human Marrow Cells
Authors: Messner H. A. , Till J. E. , McCulloch E. A.
Published: October 2019

Marrow from 28 nonleukemic individuals was separated by adherence to glass or plastic into nonadherent (NA) and adherent populations. The NA populations were found to be more dependent for colony formation in culture on added colonystimulating activity (CSA) than unseparated marrow suspensions, and therefore proved useful for CSA assays. Quantitative reconstitution procedures were used to assay CSA-producing cells. Either increasing numbers of irradiated unseparated marrow, or adherent cells derived from varying numbers of marrow cells, were used to restore colony-forming efficiency to NA populations. Assay procedures for CSA-producing cells were applied to four patients with acute leukemia prior to treatment. In all four instances, a defect in CSA-producing cells was demonstrated.
Granulocyte-Monocyte Colony-forming Capacity of Human Marrow: A Clinical Study
Authors: Craddock C. G. , Hays Esther F. , Forsén N. Rune , Rodensky Dona
Published: October 2019

A modification of the in vitro culture of marrow cells in soft agar to form granulocyte-monocyte colonies has been applied to human marrow using a constant stimulus of colony-stimulating factor (CSF) in the form of pooled serum from endotoxin-treated C57BL mice (EMS). With this system reproducible increments in colony formation were observed between two concentrations of EMS. The results indicate that the incremental response to two doses of CSF (CSF response ratio) combined with an observation of the number of colonies formed provide a more reliable measurement of colony-forming cells (CFC) than colony counts at a single dose level of CSF. The CSF response ratio in 43 marrow cultures from 34 patients with no obvious disturbance in granulopoiesis was found to be 1.65 (± 0.17 SD). Hodgkin’s disease with normal marrow showed a mean CSF response ratio of 2.04. Two subjects recovering from drug-induced agranulocytosis showed increased CSF response at a time when the blood was still neutropenic and the marrow showed hyperplasia and immaturity of the granulocytic tissue. In contrast, patients with sustained neutropenia and impaired marrow granulopoiesis showed CSF response ratios of less than 1.65. CSF response ratios in chronic myelogenous leukemia averaged 1.72, often with extremely high numbers of colonies per plate. Patients with untreated acute leukemia and marrows replaced by blasts did not form colonies. However, with partial leukemic marrow replacement, lymphoblastic leukemia marrow formed more colonies than did that of myeloblastic leukemia.
The Interaction of Hemoglobin E With β Thalassemia: A Study of Hemoglobin Synthesis in a Family of Mixed Burmese and Iranian Origin
Authors: Feldman Robert , Rieder Ronald F.
Published: October 2019

A 5-yr-old girl with hemoglobin E-β thalassemia was discovered in a family of mixed origin. The father is Iranian (β-thalassemia trait) and the mother is Burmese (hemoglobin-E trait). Hemoglobin synthesis was studied in vitro in the blood of the proposita and family members. In the subjects with hemoglobin E trait the ratio of the quantity of hemoglobin A to hemoglobin E was 3:1. However. the βA/βE synthesis ratio in reticulocytes was in the range of 1.5-2.18, and the specific activity of βE was 31%-49% greater than βA, suggesting instability of hemoglobin E with preferential destruction of abnormal hemoglobin. The blood of the proposita exhibited only hemoglobin F and hemoglobin E and reticulocytes and bone marrow showed no βA synthesis. This Iranian β-thalassemia gene is therefore of the β° type. The βE/α synthesis ratio (approximately 0.74) in blood of the proposita was similar to the βA/α ratio in mildly affected relatives with thalassemia trait. These results suggest that the severity of the hemoglobin E-β thalassemia syndrome is attributable to both instability and defective synthesis of hemoglobin E in association with absent βA synthesis due to a β° thalassemia gene.
Sensitivity of Human and Murine Hemopoietic Precursor Cells to Chemotherapeutic Agents Assessed in Cell Culture
Authors: Ogawa Makio , Bergsagel Daniel E. , McCulloch E. A.
Published: October 2019

The sensitivity of human and murine hemopoietic precursor cells to 1,3 bis (2 chloroethyl)-1-nitrosourea (BCNU), nitrogen mustard, 5-fluorouracil, and melphalan was assessed in culture using colony assay techniques. The results indicate that human and murine hemopoietic cells may differ in their intrinsic sensitivity to drugs. The hematologic toxicity of some chemotherapeutic agents cannot be extrapolated directly from mouse to man.
Cyclic Hematopoiesis in Grey Collie Dogs: A Stem-Cell Problem
Authors: Patt Harvey M. , Lund John E. , Maloney Mary A.
Published: October 2019

Studies were made to ascertain the possible basis of cyclic hematopoiesis in the grey collie. The results of 3H-thymidine and 59Fe labeling in conjunction with other marrow parameters and peripheral counts suggest that the regular periodicity of blood neutrophil and reticulocyte levels is caused by a defect at the stem-cell—marrow interface. It is postulated that the ebb and flow of hemic cell production in the grey collie reflects competition for a limiting number of pluripotential stem cells with the alternating competitive pressure provided by activation and deactivation of a neutrophil feedback circuit from periphery to stem cell. This mechanism can account for the characteristic 12-day periodicity and contributes to the phase difference in the cycling pattern of neutrophils relative to reticulocytes.
Effects of Cobalt on the Renal Erythropoietic Factor and Kidney Hydrolase Activity in the Rat
Authors: Smith Robert J. , Fisher James W.
Published: October 2019

In an experiment to determine the effects of cobalt on the renal erythropoietic factor and kidney hydrolase activity in the rat we obtained the following results: Cobalt produced significant increases in renal erythropoietic factor activity and plasma levels of erythropoietin which reached peak activity 12 hr after treatment. It also produced an increase in the activity of renal hydrolases, cathepsins A and B, which paralleled the increase in renal erythropoietic factor activity. Enzyme inhibitors which are specific for proteases, esterases, and metalloenzymes inhibited the activity of the renal erythropoietic factor in vitro. Polycythemic mice exposed to 7- and 8-day posthypoxic intervals still retained their ability to respond to in vitro generated erythropoietin when compared to mice treated on the fourth posthypoxic day. The erythropoietic activity generated by the light mitochondrial extract—normal rat serum (LME-NRS) reaction mixture was blocked by the antibody to erythropoietin. The relative concentrations of smooth and rough endoplasmic reticulum (microsomes) and vesicles (lysosomes) were approximately the same in the light mitochondrial fractions of kidneys from normal and cobalt-treated rats. Marker enzyme studies revealed primarily alkaline phosphatase activity in the light mitochondrial fraction. These studies correlate with electron micrographs of the LME which indicate a fraction composed mainly of microsomes. In addition, these data suggest a possible relationship between renal lysosomal hydrolase activity and the renal erythropoietic factor (Erythrogenin).
Erythropoiesis in Carotid Body Resected Cats
Authors: Gillis D. B. , Mitchell R. A.
Published: October 2019

It has been reported that removal of carotid bodies in cats results in a brief, profuse reticulocytosis followed by depression to below-normal levels of reticulocytes and a progressively severe anemia. Injection of cat carotid body extract was reported to increase erythrocyte 59Fe incorporation in polycythemic rats. Other studies could find no hematological abnormality in humans after bilateral carotid body resection. We reexamined the effect of carotid body resection in the cat with serial bone marrow aspirations, hematocrit determination, and reticulocyte determination. T1/2 for plasma 59Fe removal, erythrocyte 59Fe incorporation, and 51Cr lifespan determinations were performed on five operated and three control cats. No significant differences were found. We conclude that the carotid body has no direct effect on erythropoiesis and that the anemia reported in a prior study was secondary to sepsis resulting from an indwelling femoral vein catheter.
Enhanced Binding of Neuraminidase-treated Sheep Erythrocytes to Human T Lymphocytes
Authors: Weiner Michael S. , Bianco Celso , Nussenzweig Victor
Published: October 2019

Pretreatment of sheep erythrocytes with neuraminidase greatly enhances their specific binding and rosette formation with human T lymphocytes. Since the rosettes dissociate at 37°C, a method was developed to isolate human T lymphocytes with a high degree of purity and good recoveries. Trypsin treatment of E abolishes their binding to T lymphocytes. Trypsin-treated erythrocytes can be coated with antibody and complement components and used as a specific reagent to detect the C3 receptor on human B lymphocytes, avoiding the risk of an interaction between the erythrocytes and T lymphocytes.
Cell-mediated Immunity in Acute Nonlymphocytic Leukemia: Relationship to Host Factors, Therapy, and Prognosis
Authors: Greene William H. , Schimpff Stephen C. , Wiernik Peter H. , Betts Susan , Mayfield Joseph
Published: October 2019

Cell-mediated immunity in acute nonlymphocytic leukemia, either at initial presentation or in subsequent relapse, was determined among 64 patients by recall-antigen skin testing and in 37 patients by attempted sensitization with dinitrochlorobenzene (DNCB). Thirty (46.9%) of the 64 skin-test batteries displayed reactivity to at least one established antigen, and 17 (51.5%) of the 33 evaluable patients were successfully sensitized with DNCB. Twenty-six (78.8%) of the 33 patients tested with both DNCB and recall antigens reacted to either one or both. The leukemic patient’s subsequent prognosis, in terms of attainment of complete remission of his or her leukemia, duration of remission, and survival could not be correlated with the pretherapy demonstration of intact recall-antigen or DNCB cell-mediated immunity. Serial, rather than single, determinations of the state of delayed hypersensitivity may be more meaningful in terms of prognosis in patients with acute nonlymphocytic leukemia. However, increasing chemotherapeutic efficacy may obscure whatever contribution, if any, intact cell-mediated immunity makes to the eradication of malignancy.
Potential for Prolonged Disease-free Survival Following Combination Chemotherapy of Non-Hodgkin’s Lymphoma
Authors: Schein Philip S. , Chabner Bruce A. , Canellos George P. , Young Robert C. , Berard Costan , DeVita Vincent T.
Published: October 2019

The evaluation of the results of CVP and MOPP chemotherapy in 80 patients with advanced stages of non-Hodgkin’s lymphoma indicates that 36 (45%) achieved a complete remission. Twenty-eight per cent of the entire group of patients remain free of disease for periods ranging from 4 mo to over 7 yr, with a projected median duration of complete remission of 3½ yr. Significant differences in prognosis relative to histologic categories were found. Well-differentiated and nodular histology were positive determinants for improved median survival, confirming the over-all clinical validity of the Rappaport classification system for the non-Hodgkin’s lymphomas. The median survival for patients in the most clinically aggressive subgroups with diffuse histology is inferior to those with nodular patterns or well-differentiated cells. In this study it was demonstrated that it was possible to achieve a significant number of complete remissions even in the most aggressive histologic subgroups using combination chemotherapy, and these responses can be correlated with an extended disease-free survival without further therapy.
Effects of Anticoagulants and Storage on Granulocyte Function in Bank Blood
Authors: McCullough Jeffrey , Carter Sandra J. , Quie Paul G.
Published: October 2019

Storage of granulocytes for transfusion has not been practical because it has been considered that granulocyte function in bank blood is retained for only a few hours after collection. In the present study, granulocyte function was evaluated using the bactericidal assay and the quantitative nitroblue tetrazolium (NBT) method. Granulocytes from whole blood collected into acid citrate dextrose (ACD), citrate phosphate dextrose (CPD), heparin, ion exchange, and sodium citrate anticoagulants showed no functional impairment after 24 hr of storage at 4°C. With further storage, all granulocytes showed a loss of NBT activity. However, after 48 and 72 hr, granulocytes from whole blood stored in ACD and CPD killed the expected number of bacteria in the bactericidal assay. Thus, when tested in vitro, granulocytes maintain normal function, at least during the first 24 hr after collection when stored in certain anticoagulants under standard blood bank conditions.
An Elution Procedure for Visualization of Adult Hemoglobins in Human Blood Smears
Authors: Kabat David
Published: October 2019

A procedure is described for visualization of normal and mutant adult hemoglobins in human blood smears. After extraction of blood smears with a concentrated potassium phosphate buffer (2.76 M, pH 7.2), erythrocytes that had adult hemoglobins stained bright red with erythrosin, whereas cells that had only fetal hemoglobin appeared as clear ghosts. Analyses of cord blood from newborn infants indicate that, although most erythrocytes contain only Hb F and a few contain only Hb A, many contain both hemoglobins A and F.
G-6-PD Manchester: A New Variant Associated With Chronic Nonspherocytic Hemolytic Anemia
Authors: Milner Gillian , Delamore Irvine W. , Yoshida Akira
Published: October 2019

A variant of glucose-6-phosphate dehydrogenase (G-6-PD) designated G-6-PD Manchester, and associated with chronic nonspherocytic hemolytic anemia, was found in an English male. The electrophoretic mobility at pH 7.0 of this G-6-PD variant is the slowest yet described. Substrate specificity is normal but enzyme activity is markedly inhibited by NADPH, and this is thought to account for the severe chronic hemolysis.
The Plasminogen Activator of Vampire Bat Saliva
Authors: Cartwright Terence
Published: October 2019

A procedure is described for the production of a highly active plasminogen activator (desmokinase) from the saliva of the vampire bat using gel filtration on Biogel A 0.5 m followed by ion-exchange chromatography on DEAE-cellulose. Investigations using specific inhibitors indicated that desmokinase was probably a specific protease similar in some respect to urokinase. Desmokinase was not, however, able to activate plasminogen in solution, and it is postulated that this could be due to a requirement for a fibrin surface for activation to proceed. Desmokinase seems to be more effective than either urokinase or streptokinase in the lysis of preformed blood clots. This property may be explicable in terms of greater affinity of desmokinase for fibrin.
In Vitro Growth of Granulocytic Colonies From Circulating Cells in Human Cord Blood
Authors: Knudtzon Søren
Published: October 2019

Human umbilical cord blood cells from 26 newborn infants and peripheral blood cells from 18 adults were cultured in vitro by using the agar-gel method of human hemopoietic cell culture. An increased concentration of colony-forming cells was seen in the cord blood cultures. Between 17 and 385 colonies, with a mean of 122, were formed in these cultures per 2 x 105 nucleated cells plated. The peripheral blood cell cultures from adults gave rise to 0-11 colonies, with a mean of 3, per 2 x 105 nucleated cells plated. The average number of cells per colony was 1000-1500 cells after 14 days of culture, predominantly granulocytic.
Cyclical Granulopoiesis in Chronic Granulocytic Leukemia: A Simulation Study
Authors: Wheldon T. E. , Kirk J. , Finlay Helen M.
Published: October 2019

There exists ample evidence that granulopoiesis is an actively controlled process. The observation of cyclical granulopoiesis in chronic granulocytic leukemia (CGL) suggests that control is deranged rather than abolished in this disorder. Analysis of the kinetics of granulopoiesis in CGL may provide some insight into the nature of the derangement. To facilitate analysis, a mathematical model of the granulopoietic control system is proposed and examined using computer simulation. With model parameters initially chosen to represent normal granulopoiesis, the minimal changes necessary to represent granulopoiesis in CGL were investigated. This analysis indicates that two separate changes seem to be required: (1) the granulocyte maturation time must be increased and (2) the precursor input to the granulocytic pathway of development must also be increased. A causal association between delayed maturation and rising precursor input is suggested, and some possible mechanisms of association are proposed.
N-Acetylneuraminic Acid Deficiency in Erythrocyte Membranes: Biophysical and Biochemical Correlates
Authors: Chien Shu , Cooper George W. , Jan Kung-ming , Miller Louis H. , Howe Calderon , Usami Shunichi , Lalezari Parviz
Published: October 2019

The electrochemical and rheologic behaviors of red blood cells (RBCs) from a patient with acquired polyagglutinability were studied. The N-acetylneuraminic acid content of the RBCs averaged less than one-half normal. The majority of the patient’s RBCs (over 90%) showed a reduction in electrophoretic mobility and a decrease in binding of positively charged colloid with a widening of the charge-free areas, but a small population exhibited normal surface charge density. Dextran 40, which does not induce significant aggregation of normal RBCs, caused the patient’s RBCs to form rouleaux. Dextran 80 induced a stronger aggregation of the patient’s RBCs than that of normal RBCs. The increased tendency of the patient’s RBCs to be aggregated by macromolecules was associated with an elevation of blood viscosity. This study on a clinical patient has provided evidence for the conclusions previously derived from experimental studies that the stability of RBC aggregation depends upon a balance between macromolecular bridging force and electrostatic repulsive force and that alterations in RBC surface charge may affect cell aggregation and blood viscosity.
Trisomy 8 in Acute Myeloblastic Leukemia and Sideroachrestic Anemia
Authors: Jonasson J. , Gahrton G. , Lindsten J. , Simonsson-Lindemalm C. , Zech L.
Published: October 2019

An extra chromosome number 8 was found in the bone marrow of one patient with acute myeloblastic leukemia and one with sideroachrestic anemia. The extra chromosome was not found in the peripheral blood of any of the patients. In remission after treatment with cytotoxic drugs, the patient with acute myeloblastic leukemia had normal 46 XX cells in the bone marrow. In relapse, the same patient demonstrated two reciprocal translocations in addition to trisomy 8. This was interpreted as evidence for successive steps of clonal evolution from a single ancestor cell.
Separation of Megakaryocytes From Mouse Bone Marrow by Velocity Sedimentation
Authors: Nakeff A. , Maat B.
Published: October 2019

A technique for separating mouse bone marrow megakaryocytes by unit gravity velocity sedimentation is described using a modified sedimentation chamber. The velocity sedimentation profile for megakaryocytes comprises three peaks corresponding to sedimentation velocities of 30, 60, and 100 mm/hr. Morphologic examination of megakaryocytes in the 30 mm/hr peak reveals them to be primarily immature with the latter two peaks corresponding to larger, more mature megakaryocytes.