bluuebunny/crossref_metadata_2025 huggingface dataset:
Blood group D antigen content of nucleated red cell precursors
Authors: Rearden A , Masouredis SP
Published: October 2019

The D antigen content of nucleated red cell precursors in human bone marrow was estimated using autoradiography and 125I-anti-D. D antigen first appeared in the pronormoblast, and the quantity of antigen progressively increased during red cell maturation. Maximal anti-D binding occurred on mature red blood cells. Pronormoblasts, basophilic normoblasts, polychromatophilic normoblasts, and orthochromatic normoblasts, respectively, had approximately 1/4, 1/2, 2/3, and 3/4 the quantity of antigen found on mature red cells. None of the other cell types were found in bone marrow labeled with anti-D.
Erythropoiesis during recovery from macrocytic anemia: macrocytes, normocytes, and microcytes
Authors: Bessman D
Published: October 2019

In seven patients with marked megaloblastic anemia (MCV greater than 110 fl), red cell size distribution curves (erythrograms) demonstrated the size of red cells produced after therapy. In six, the new red cells were normocytic throughout recovery. In the seventh patient, folate repletion along produced a new population of microcytes, due to unsuspected iron deficiency; after iron repletion normocytes were produced. Three patients with autoimmune hemolytic anemia had macrocytosis (MCV greater than 110 fl) without folate or vitamin B12 deficiency. During recovery with predisone therapy, instead of a discrete new normocytic population appearing, the entire population progressively returned to normal size. Normal rather than “stress” reticulocytes, and remodeled stress reticulocytes remaining, may explain this different pattern of recovery. Two patients initially had minor subpopulations of smaller red cells that disappeared soon after therapy. These probably reflected the dyserythropoiesis of severe megaloblastic anemia.
An unusual case of a plasma cell neoplasm with an IgG3lambda myeloma and a gamma3 heavy chain disease protein
Authors: Adlersberg JB , Grann V , Zucker-Franklin D , Frangione B , Franklin EC
Published: October 2019

A unique case of gamma3 heavy chain disease with two related serum proteins is reported. One molecule appears to be an IgG3lambda myeloma protein. The second molecule is a dimer of a shortened gamma3 heavy chain that has an unblocked amino terminus and lacks the VH and CH1 domains. Its probable origin as a synthetic product is discussed. The clinical and pathologic features of this patient resemble those of other patients with gamma heavy chain disease more than those of patients with multiple myeloma. It seems likely that the heavy chain disease protein is the result of a mutational event in the malignant clone originally producing the myeloma protein.
Antibody-dependent lymphocyte-mediated granulocyte cytotoxicity in man
Authors: Logue GL , Kurlander R , Pepe P , Davis W , Silberman H
Published: October 2019

Sera from two patients with granulocytopenia associated with collagen vascular disease caused the destruction of normal human granulocytes by autologous lymphocytes in vitro. Granulocyte cytotoxicity was measured by the release of 51Cr during incubation with test sera and lymphocytes in microtiter plates. Between 8% and 46% granulocytoxicity was produced in granulocytes from 8 normal donors by the sera from these two patients. Less than 6% granulocytotoxicity was seen with the sera from 14 normal subjects and 29 patient controls. Treatment of lymphocyte preparations with carbonyl iron and magnetic separation to remove phagocytic cells or treatment with complement-coated red cells followed by repeated gradient centrifugation to remove complement receptor- bearing lymphocytes did not reduce the granulocytotoxicity. There was a dose-response relationship between the concentration of positive sera and granulocytotoxicity. When these sera were fractionated by Sephadex G-200 gel filtration and by ion-exchange chromatography with DEAE- cellulose, the active component appeared in the IgG-containing fractions. Thus, IgG antibody-dependent, lymphocyte-mediated granulocyte cytotoxicity represents a means of detecting human granulocyte antibodies and is a possible mechanism of autoimmune neutropenia in these two patients.
Terminal deoxyribonucleotidyl transferase activity in B-cell acute lymphocytic leukemia
Authors: Shaw MT , Dwyer JM , Allaudeen HS , Weitzman HA
Published: October 2019

Moderately high levels of activity of the enzyme terminal deoxyribonucleotidyl transferase (TdT) were found in the leukemic cells of a patient with acute lymphyocytic leukemia. The proliferating cells were B lymphocytes bearing IgG antibody, and the disease was associated with an IgG monoclonal spike and a mediastinal mass. The observations in this case suggest that TdT is related more to the immaturity and proliferation of certain lymphoid stem cells than to their progress toward B- or T-cell differentiation.
Differences in the terminal steps of complement lysis of normal and paroxysmal nocturnal hemoglobinuria red cells
Authors: Rouault TA , Rosse WF , Bell S , Shelburne J
Published: October 2019

The number of microscopically visible lesions produced on the membrane for a given degree of lysis on normal cells as on PNH cells. Since complement lesions were not formed until C8 or C9 was incorporated into the complement sequence, the results suggest that increased lysis of red cells in PNH is due at least in part to more efficient penetration of the PNH membrane by the terminal lytic sequence of complement. Furthermore, the efficiency of the terminal lytic sequence in the lysis of PNH cells when complement was activated by the alternative pathway and the classical pathway was analyzed. There was no significant difference (p less than 0.01) in the number of lesion present at an equivalent degree of lysis when initiated by antibody, cobra venom factor, or acidification. Thus, the efficiency of the terminal lytic sequence does not vary with different modes of activation.
Multimarker analysis of T-cell chronic lymphocytic leukemia
Authors: Marks SM , Yanovich S , Rosenthal DS , Moloney WC , Schlossman SF
Published: October 2019

A 68-yr-old male with chronic lymphocytic leukemia (CLL) presented with splenomegaly and skin infiltration but no lymphadenopathy. The peripheral blood WBC cound was 300 x 10(9)/liter, with 95% small mature- appearing lymphocytes that were E-rosette positive and EAC-rosette negative. Further characterization of the patient's cells was performed using antisera with known lymphoid sub-population specificity. Anti- p23,30, which reacts with normal circulating B cells but not with T cells or thymocytes, was unreactive with the patient's cells. Anti-311, which reacts with both thymocytes and circulating T cells, was reactive with the patient's cells. Anti-Bk, which reacts only with thymocytes and not with circulating T-cells, failed to react with the patient's cells. The enzyme terminal deoxynucleotidyl transferase, present in thymocytes but absent for circulating T-cells, was also absent from the patient's lymphoid cells. Multimarker analysis therefore showed a mature T-lymphocyte phenotype on this patient's leukemia cells. Further functional analysis will probably show that such cells represent clonal expansion of a mature T-cell subpopulation, analogous to the B-cell clonality of common-variant CLL.
Human cell lines that elaborate colon-stimulating activity for the marrow cells of man and other species
Authors: Di Persio JF , Brennan JK , Lichtman MA , Speiser BL
Published: October 2019

We have established two human cell lines which elaborate colony- stimulating activity (CSA) for at least four species: man, mouse, rabbit, and dog. One, GCT, was isolated from a lung metastasis of a fibrous histiocytoma; the other, RC4, from a monocyte-enriched fraction of normal blood. Medium conditioned by either GCT or RC4 cells was more potent in stimulating human marrow growth in vitro than was monocyte-conditioned medium or human leukocyte feeder layers. Fractionation of cell-line-conditioned medium by Sephacryl S-200 chromatography indicated that the maximum activity of the CSA for human marrow cells is eluted within the range of 30,000–40,000 daltons. These cells lines provide a continuous source of large quantities of conditioned medium for purification of CSA. Moreover, the invariable growth-supporting activity for all species tested and the high potency of cell-line CSA facilitates studies of its elaboration and biologic effects.
Fetal erythropoiesis in steel mutant mice. III. Defect in differentiation from BFU-E to CFU-E during early development
Authors: Chui DH , Liao SK , Walker K
Published: October 2019

Erythroid progenitor cells in +/+ and Sl/Sld fetal livers manifested as burst-forming units-erythroid (BFU-E) and colony-forming units- erythroid (CFU-E) were assayed in vitro during early development. The proportion of BFU-E was higher as mutant than in normal fetal livers. On the other hand, the proportion of CFU-E was less in the mutant than in the normal. These results suggest that the defect in Sl/Sld fetal hepatic erythropoiesis is expressed at the steps of differentiation that effect the transition from BFU-E to CFU-E.
Treatment of chronic granulocytic leukemia with melphalan
Authors: Hauch T , Logue G , Laszlo J , Cox E , Rundles W
Published: October 2019

Thirty-three patients with newly diagnosed chronic granulocytic leukemia (CGL) were treated with melphalan between 1968 and 1976. Within 3 mo of beginning therapy subjective and objective disease parameters improved. Disease control was easily maintained with this agent until hematologic exacerbation occurred. The median duration of disease control was 25.3 mo, and the median duration of survival was 28.6 mo. Serious side effects were not produced. Thus melphalan appears to be another agent that may be used to control the manifestations of CGL prior to hematologic exacerbation.
Three-dimensional model of bone marrow
Authors: Mohandas N , Prenant M
Published: October 2019

Three-dimensional scale models of bone marrow from a hypertransfused and a normal rat were constructed. The model of marrow from the hypertransfused rat demonstrated the existence of distinct erythroblastic islands in situ in which the erythroblasts underwent sychronous maturation. Macrophages were found in close association with the developing erythroblasts. The immature erythroblasts were tightly grouped, but as they matured they began to move apart. Erythroblasts in individual clusters were found to be at the same stage of morphologic maturation. In contrast, the model of marrow from the normal rat showed a majority of clusters containing erythroblasts at various stages of maturation. Erythropoiesis was not spatially restricted to the area proximal to the sinuses but was found to occur over the entire marrow space. Thrombopoiesis, however, was found to take place exclusively in the immediate vicinity of the marrow sinuses.
Platelet factor VIII-related antigen: studies in vivo after transfusion in patients with von Willebrand disease
Authors: Sultan Y , Jeanneau C , Lamaziere J , Maisonneuve P , Caen JP
Published: October 2019

Four unrelated patients with a severe form of von Willebrand disease showed no detectable factor VIII-related antigen (VIIIR:AG) in either their plasma or their platelets. They received cryoprecipitate infusions, three patients in a single injection each and one every day for 9 days before and after surgery. Platelet VIIIR:Ag was studied at different times during and after transfusion using electroimmunoassay of platelet extracts and electron microscopy of the platelets incubated with anti-VIIIR:Ag antibodies coupled to peroxidase. No VIIIR-Ag was detected in or around the patients' platelets, although this antigen was detected in the circulating blood. These results that there was no VIIIR:Ag uptake from the plasma by the platelets and that platelet VIIIR:Ag came from megakaryocytes.
"Concentration x time" methotrexate via a subcutaneous reservoir: a less toxic regimen for intraventricular chemotherapy of central nervous system neoplasms
Authors: Bleyer WA , Poplack DG , Simon RM
Published: October 2019

Neurotoxicity associated with intrathecal methotrexate therapy has been shown to correlate with elevated concentrations of the drug in the cerebrospinal fluid as well as with the total cumulative dosage. In our study 19 patients with meningeal leukemia were randomized to receive courses of intraventricular methotrexate via an Ommaya reservoir consisting of either single injections of 12 mg/sq m/dose or a low-dose “concentration x time” (C x T) schedule of 1 mg every 12 hr for 3 days. There were no significant differences between the two treatment groups in the rate of remission induction, the number of relapses, or the durations of remission. The mean (+/- 1 SD) cumulative methotrexate dose was 66 +/- 41 mg/sq m in the C x T group and 173 +/- 64 mg/sq m in the 12 mg/sq m/dose group (p less than 0.005). Neurologic toxicity occurred in one of the eight patients in the C x T group and in seven of ten patients in the 12 mg/sq m/dose group (p less than 0.05). These observations suggest that the C x T dosage schedule is less neurotoxic and equally effective in the treatment of central nervous system leukemia.
Granulocyte modulation of endotoxin-stimulated colony-stimulating activity (CSA) production
Authors: Mahmood T , Robinson WA
Published: October 2019

Incubation of human macrophages with endotoxin resulted in significantly enhanced colony-stimulating activity (CSA) production by these cells. Preincubation of endotoxin with mature granulocytes abolished this stimulatory effect. The stimulatory effect of endotoxin on macrophage CSA production was not abolished, however, by preincubation with NaF-treated granulocytes, granulocyte membranes, or nonphagocytic cells (lymphocytes or erythrocytes). These data suggest that mature granulocytes may play a role in the modulation of CSA production and granulopoiesis by inactivation of stimulatory materials such as endotoxin.
Effects of actinomycin D in vivo on murine erythroid stem cells
Authors: Zuckerman KS , Sullivan R , Quesenberry PJ
Published: October 2019

Low-dose actinomycin D (Acto) selectively suppresses murine erythropoiesis without decreasing erythropoietin (Ep) production. We used the plasma clot system to determine the stage of erythroid differentiation at which this inhibition occurs. Late erythroid precursors, CFU-E, and less differentiated committed erythroid stem cells, BFU-E, were assayed in CF1 mice given Acto 75–82 microgram/kg/day or saline subcutaneously for 5 days. We also assayed pluripotent (CFU-S) and committed granulocyte-monocyte (CFU-C) stem cells. Reticulocytes and marrow and spleen nucleated erythroid precursors were decreased by 99% in the Acto-treated mice; tibial marrow CFU-E were decreased by 97% and splenic CFU-E by 99%. Tibial BFU- E were not decreased by Acto, although there was a 66% diminution in splenic BFU-E. Acto increased tibial CFU-S, but splenic CFU-S and tibial and splenic CFU-C were unchanged. Thus Acto inhibits erythropoiesis by suppressing the ability of immediate committed erythroid precursors of CFU-E or CFU-E themselves to differentiate further in response to Ep. Acto does not affect survival or proliferation of the less differentiated cells--CFU-C, CFU-S, and marrow BFU-E. The suppression of splenic BFU-E in Acto-treated mice may indicate that marrow and splenic BFU-E are basically different stem cells. Alternatively, Acto treatment may impair migration of BFU-E from marrow to spleen.
Diamond-Blackfan syndrome: evidence against cell-mediated erythropoietic suppression
Authors: Freedman MH , Saunders EF
Published: October 2019

The profound anemia of Diamond-Blackfan syndrome (DBS) is due to marrow red cell failure, but the pathogenesis is not understood. Studies by others indicated cell-mediated erythropoietic suppression in this condition. To explore this mechanism further, Ficoll-Hypaque--separated peripheral blood lymphocytes (PBL) from four anemic untreated patients with DBS, or from normals were cocultured with control marrow in vitro and the growth of erythropoietin-responsive stem cell colonies (CFU-E) was dermined. CFU-E numbers obtained from cultures with added normal PBL were not significantly different from the number without PBL. Similarly, CFU-E from cultures with added DBS PBL were not significantly different from the number without PBL (215 versus 220, 229 versus 220 and 84 versus 60, 74 versus 94/10(5) cells, respectively). Mixing marrows from a control and one DBS patient in ratios of 2:1, 1:1, or 1:2 prior to culture failed to disclose a decrease of colony growth. We could not show cellular inhibition of erythropoiesis in these patients with DBS. The mechanism of anemia in this disorder remains an open question.
Separation of rabbit marrow precursor cells by combined isopycnic sedimentation and electronic cell sorting
Authors: Scott RB , Grogan WM , Collins JM
Published: October 2019

Separation of developing cells into fractions of differing stages of maturity is critical to effective biochemical study of the process of cellular differentiation. Density gradient techniques utilizing rate- zonal or isopycnic separations have permitted partial separations based on cell mass or cell density. In this study the separation of various rabbit marrow cells was improved by high-speed flow analysis and sorting in a Coulter Two-Parameter Cell Sorter. After preliminary isopycnic separation of marrow cells in Ficoll-Hypaque, cells were sorted into lymphoid and myeloid elements, utilizing light-scatter (LS) profiles to determine sorting. Characteristic LS patterns were present for erythrocytes, lymphocytes, devitalized cells, and granulocytes. When erythrocytes and their precursors were removed by hypotonic lysis, the remaining granulocytes could be sorted to give samples with much greater purity with respect to developmental stage than was possible with density gradients alone. Thus the combination of techniques represents a significant improvement in the ability to study the mechanisms of normal or altered cellular maturation.
Monovalent cation composition and ATP and lipid content of irreversibly sickled cells
Authors: Clark MR , Unger RC , Shohet SB
Published: October 2019

Discontinuous Stractan gradients were used to separate heterogeneous populations of sickle cells into discrete subpopulations containing varying proportions of reticulocytes, mature discoid cells, and irreversibly sickled cells (ISC). The improved homogeneity of these preparations, together with an enhanced yield of ISC, allowed us to distinguish effects of cell maturation from those of irreversible sickling. With these cell preparations we have begun to define physical properties of ISC. We confirmed the marked abnormalities in cation composition of native ISC. Measurements of ATP in ISC did not substantiate prior reports of ATP deficits. Finally, no evidence for substantial loss of membrane lipids during the process of ISC formation could be demonstrated.
Regulation of proliferation of murine megakaryocyte progenitor cells by cell cycle
Authors: Williams N , Jackson H
Published: October 2019

The extent to which mouse megakaryocyte progenitor cells (colony- forming unit-megakaryocyte, CFU-M) can proliferate in semisolid cultures prior to endomitosis, and conditions that may regulate that differentiation step, were investigated. The proliferative capacity of CFU-M was determined by estimating the number of megakaryocytes per colony. A bimodal distribution was observed (modal values, 10–15 and 25- 30 cells/colony), indicating that separate megakaryocyte progenitor cells may be biased in their capacity for proliferation versus endomitosis. Differences were observed in the cell cycle characteristics of CFU-M as determined in vivo and in vitro that suggest that maturation of CFU-M into megakaryocytes may be regulated within the marrow by control of the cell cycle of the megakaryocyte precursor cell.
Red cell 2,3-diphosphoglycerate and oxygen affinity of hemoglobin in patients with thyroid disorders
Authors: Zaroulis CG , Kourides IA , Valeri CR
Published: October 2019

We measured red blood cell 2,3-diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), and the P50 value in vitro of the oxyhemoglobin dissociation curve, which is the oxygen tension at half saturation of hemoglobin, in order to quantitate red blood cell oxygen transport function in individuals who were diagnosed as hypothyroid, euthyroid, or hyperthyroid based on measurements of thyroxine (T4), triiodothyronine (T3), thyrotropin (TSH), and their clinical status. Hypothyroid (mean T4 2.8 microgram/dl, T3 49 ng/dl, TSH 37 microU/ml) and hyperthyroid (mean T4 14 microgram/dl, T3 271 ng/dl, TSH less than 0.7 microU/ml) patients had normal red cell 2,3-DPG and ATP levels and normal P50 values in vitro. The known changes in oxygen consumption produced by alterations in thyroid hormone levels in patients with hypothyroidism or hyperthyroidism did not affect red blood cell oxygen transport function.
Surface glycoproteins of human white blood cells. Analysis by surface labeling
Authors: Andersson LC , Gahmberg CG
Published: October 2019

We labeled surface glycoproteins of normal human blood platelets, granulocytes, monocytes, T and B lymphocytes, and null cells by the galactose oxidase-NaB3H4 and periodate-NaB3H4 labeling techniques. The labeled glycoproteins were observed by fluorography after separation by polyacrylamide slab gel electrophoresis. All major types of human leukocytes showed different and characteristic surface glycoprotein patterns. These patterns evidently also include common components.
Pokeweed mitogen-induced B cell differentiation in idiopathic aplastic anemia associated with hypogammaglobulinemia
Authors: Uchiyama T , Nagai K , Yamagishi M , Takatsuki K , Uchino H
Published: October 2019

Peripheral blood lymphocytes from 14 patients with idiopathic aplastic anemia were examined to determine their capacity to differentiate into immunoglobulin (lg) producing cells in the presence of pokeweed mitogen (PWM). Four patients had hypogammaglobulinemia. Lymphocytes from ten patients without and one with hypogammaglobulinemia were capable of differentiating into lg-producing cells. Lymphocytes from three patients with hypogammaglobulinemia, however, were unable to differentiate. Coculture experiments showed that in one patient helper activity of T cells was impaired, in another patient B cells could not be stimulated by PWM to differentiate even in the presence of normal T cells, and in a third patient both the helper function of T cells and the capacity of B cells to differentiate into lg-producing cells were affected. These results suggest that differentiation of lymphoid cells is impaired in some cases of idiopathic aplastic anemia associated with hypogammaglobulinemia.
Monocyte-mediated inhibition of lymphocyte blastogenesis in Hodgkin disease
Authors: Schechter GP , Soehnlen F
Published: October 2019

Mononuclear leukocytes isloated from the blood of previously treated patients with advanced active Hodgkin disease contained high concentrations of monocytes and showed poor lymphocyte blastogenesis to mitogens. In five of eight patients with disseminated disease, blastogenesis became normal or improved markedly when the leukocyte suspensions were depleted of monocytes before culture. Addition of autologous macrophages to the monocyte-depleted lymphocytes resulted in a reappearance of the inhibition of blastogenesis. Monocyte inhibition was associated with the presence of active disease, lymphocytopenia, and low lymphocyte/monocyte ratios in the peripheral blood. The role of previous treatment is uncertain, since inhibition tended to disappear when the patients were retreated. Inhibitory monocyte-lymphocyte interactions may be one of the causes of impaired cell-mediated immunity in Hodgkin disease.
Platelet storage at 22 degrees C: effect of type of agitation on morphology, viability, and function in vitro
Authors: Holme S , Vaidja K , Murphy S
Published: October 2019

Recovery in vivo after 51Cr labeling, platelet morphology, and platelet aggregation were studied with platelet concentrates (PC) stored for transfusion under carefully controlled conditions. PC were prepared to a final volume of 50 ml from whole blood anticoagulated with citrate- phosphate-dextrose (CPD). The platelet count was kept between 0.8 and 1.6 X 10(12) platelets/liter. The PC were stored in bags constructed of polyvinylchloride (PVC) or polyethylene (PE) at 22 degrees C for 72 hr. The bags were placed on a horizontal shaker or a ferris wheel for agitation during storage. No significant changes in pH or platelet count were observed during storage. PC stored on the wheel showed moderate loss of viability and a marked deterioration of platelet morphology and aggregation compared to the shaker. PC stored on the shaker in bags made of PE showed better aggregation with ADP and thrombin but had the same viability and morphology as PC in bags constructed of PVC. Maintenance of normal platelet morphology as determined by phase-contrast microscopy, extent of shape change response, and the size distribution according to the Coulter Counter correlated with recovery in vivo.
Nonreversible loss of platelet aggregability induced by calcium deprivation
Authors: Zucker MB , Grant RA
Published: October 2019

Platelets lose their ability to aggregate when deprived of divalent cations. This usually was studied by incubating human citrated platelet- rich plasma with EDTA or EGTA and then adding enough CaCl2 to combine with the chelating agent. Incubation for 5–7 min at 37 degrees C caused irreversible loss of the platelets' ability to adhere to glass and to aggregate with ADP, epinephrine, A23187, vasopressin, or serotonin or upon rewarming after chilling and markedly reduced aggregation with collagen or thrombin. Control samples incubated with saline, CaEDTA, or CaEGTA were not inhibited. Untreated platelets washed and incubated in solutions treated with resins that remove divalent cations lost their ability to aggregate in 30 min. More than about 0.26 mM Mg2+ partially protected the platelets. Unlike aggregation, ADP-induced shape change, clot retraction caused by thrombin or ADP plus reptilase, and thrombin- induced 14C-serotonin release were not inhibited after incubation. Aggregability was not restored by prolonged incubation with CaCl2, adding normal plasma, or washing the platelets. Its loss was temperature and pH dependent, occurring in 2 min at 43 degrees C but not in 7 min at 30 degrees C, and at pH 7.8 but much less at pH 7.2. The defect was not associated with an increase in platelet cyclic AMP, a decrease in metabolic ATP, or the presence of free ADP.
Effect of acute alcohol intoxication on granulocyte mobilization and kinetics
Authors: Gluckman SJ , MacGregor RR
Published: October 2019

Granulocyte mobilization into skin abrasions in human volunteers was significantly inhibited by acute alcohol intoxication (45,-800 cells in 8 hr versus 353,000 in normal controls). Alcohol applied locally did not inhibit granulocyte delivery, and protection of the abrasion against heat loss did not reduce the inhibited delivery in intoxicated volunteers. Intoxication inhibited granulocyte adherence and local mobilization in parallel. Alcohol administration to rabbits shifted granulocytes from marginal to circulating pool in a manner similar to epinephrine. Mobilization of bone marrow granulocytes by glucocorticoid or endotoxin administration was not inhibited by intoxication, nor did it prevent the endotoxin-induced shift of granulocytes from circulating to marginal pool.
Fragmentation and myelin formation in hereditary xerocytosis and other hemolytic anemias
Authors: Snyder LM , Lutz HU , Sauberman N , Jacobs J , Fortier NL
Published: October 2019

Erythrocytes from a heterogeneous group of hemolytic anemias have been found to release acetylcholinesterase-enriched fragments and show myelin forms during ATP depletion in vitro. The highest amount of fragmentation was found in hereditary spherocytosis and xerocytosis, two inherited membrane defects. Our data suggest ATP depletion plays a role in producing fragmentation or myelin forms. The addition of external CaCl2 1 mM had no effect on the degree of fragmentation. However, propranolol hydrochloride, a cationic anesthetic that does not prevent ATP depletion, inhibited fragmentation and the appearance of myelin forms in both hereditary spherocytes and xerocytes. A more detailed study of the xerocyte fragments showed that they had the same protein composition as those from normal red cells, primarily integral membrane proteins and glycoproteins. The red cells from patients with PNH and G6PD deficiency had the shortest survival in vivo (51Cr) and produced the smallest amount of fragmentation and myelin forms in vitro, whereas xerocytosis with only mild to moderate hemolysis in vivo was associated with the highest amount of myelin forms and membrane fragments in vitro.
Purification and some properties of colony-stimulating factor from normal human urine
Authors: Motoyoshi K , Takaku F , Mizoguchi H , Miura Y
Published: October 2019

A colony-stimulating factor (CSF) that stimulated human and mouse bone marrow cells to proliferate in vitro and form pure granuloid colonies was purified about 4000-fold from normal human urine. Purification procedures included concentration with polyethyleneglycol, ammonium sulfate precipitation, two chromatographic separations on DEAE- cellulose columns, gel filtration, and polyacrylamide gel electrophoresis. The molecular weight of the purified factor was estimated to be about 85,000 daltons by gel filtration, and the specific activity was found to be 10(6) or 6.7 X 10(5) colonies/mg protein using mouse or human bone marrow cells, respectively. A urinary colony-inhibiting factor was separated from the CSF on the first DEAE- cellulose column. This inhibitor suppressed the formation of pure granuloid colonies of human and mouse bone marrow cells when employed in conjunction with the purified urinary CSF.
Acute lymphoblastic leukemia followed by chronic myelocytic leukemia
Authors: Tosato G , Whang-Peng J , Levine AS , Poplack DG
Published: October 2019

Second hematologic malignancies occur rarely in patients previously treated for leukemia. This report describes a patient with acute lymphoblastic leukemia who remained in complete remission for 5 yr and then developed chronic myelocytic leukemia (CML). The original lymphoblasts were associated with a partial deletion of chromosome 21, while CML was associated with a classic Philadelphia marker, indicating the independent origin of the two leukemias.
Variable globin chain synthesis in mouse erythroleukemia cells
Authors: Alter BP , Goff SC
Published: October 2019

Various mouse erythroleukemia cell lines show unique responses to chemical inducers. Lines 745 and 707, from DBA/2 mice, produced 25%-- 48% beta-minor and 78%--52% beta-major globin following culture with DMSO. Butyric acid treatment led to approximately 40% beta-minor globin, while induction with hemin resulted in over 80% beta-minor synthesis. Line FSD was developed independently. DMSO induction led to less than 10% beta-minor globin, while hemin and butyric acid both resulted in approximately 40% beta-minor synthesis. T3C12 and 5000 originated in DDD mice. With these lines all inducers led to only beta- major and no beta-minor synthesis. The inducers did not influence protein stability or initiation of globin synthesis. Translation of poly A-containing cytoplasmic RNA in a wheat germ cell-free system led to proportions of beta-minor that were the same as those seen in the intact cells. Thus regulation of the type of globin chain produced in erythroleukemia cells following culture with several inducing agents occurs at either the level of gene transcription or posttranslational processing, or by growth of a selected cell population.
Purine nucleoside phosphorylase in chronic lymphocytic leukemia (CLL)
Authors: Borgers M , Verhaegen H , De Brabander M , De Cree J , De Cock W , Thone F , Geuens G
Published: October 2019

Purine nucleoside phosphorylase (PNP), the enzyme schematically next to adenosine deaminase in the purine salvage pathway, has been demonstrated cytochemically in peripheral blood lymphocytes of healthy subjects and chronic lymphocytic leukemia (CLL) patients. The enzyme activity is confined to the cytosol. In healthy subjects the majority of lymphocytes are strongly reactive for PNP, whereas the rest are devoid of cytochemically demonstrable activity. The percentage of PNP- positive cells largely corresponds to the number of E rosette-forming cells and is inversely proportional to the number of Ig-bearing cells. In six of seven CLL patients studied only a minor percentage of the lymphocytes showed strong PNP activity, whereas the large majority (88%- -98%) possessed trace activity. Such patients have a high number of Ig- bearing cells and a low number of E rosette-forming cells. A different pattern of markers was found in the lymphocytes of the seventh CLL patient: 66% were strongly reactive for PNP, an important number formed E rosettes, and a minor percentage were Ig bearing. These data indicate that PNP can be useful as a “nonmembrane” marker in the differentiation of the B and T cell origin in CLL and deserves to be studied in other lymphoproliferative disorders.
Coupled amidolytic assay for factor VII: its use with a clotting assay to determine the activity state of factor VII
Authors: Seligsohn U , Osterud B , Rapaport SI
Published: October 2019

A coupled amidolytic assay for factor VII (VII) has been developed that when used with a clotting assay for VII enables detection of activated VII. In the assay, VII in a test material determines generation of factor Xa in a mixture of purified factor X, tissue factor, and calcium; factor Xa is measured with a chromogenic substrate. Factor VII activity in the coupled amidolytic assay (VIIam) correlated well with VII activity in a one-stage clotting assay (VIIc) in 57 healthy subjects, 5 patients with hereditary VII deficiency, and 11 patients with liver disease. Activation of plasma VII by kaolin, clotting, or cold strikingly increased VIIc but not VIIam levels. Thus the ratio VIIc/VIIam (VII activity ratio) is a measure of VII activation. In 27 warfarin-treated patients the mean VII activity ratio was significantly decreased, reflecting a greater decline in VIIc than in VIIam. This probably stems from partially carboxylated VII being able to act during the 3-min incubation of the amidolytic assay but unable to act rapidly enough to affect the clotting assay. Measurement of VIIc/VIIam should enable evaluation of the activity state of VII in thrombotic disorders and in components for transfusion therapy.
Banding studies on chromosomes in diffuse "histiocytic" lymphomas: correlation of 14q+ marker chromosome with cytology
Authors: Fukuhara S , Rowley JD , Variakojis D , Sweet DL Jr
Published: October 2019

Chromosomes were studied in cells from tissues primarily involved by diffuse “histiocytic” lymphoma in nine patients. Two of the patients had stage II disease; their tumors were fibrotic and had no mitotic cells. One patient was in stage III, and the remaining six patients had stage IV disease. The modal chromosome number of abnormal cells from these last seven patients was hypodiploid in two, hyperdiploid in four, and near-triploid in one. Complete banding studies of six cases and partial analysis of the seventh indicate that (1) every patient had a distinct cell line with common markers, with a few cells showing minor variants; (2) although certain chromosomes (Nos. 1, 2, 3, 9, 12, and 14) were structurally affected more often than others, no markers with the same banding pattern were noted among them; and (3) the cytologic type of lymphoma could be correlated with the karyotype in all seven patients. When the Lukes and Collins classification was used, three patients whose tumors were composed predominantly of large noncleaved cells showed a 14q translocation leading to the formation of a 14q+ marker chromosome. This marker was not observed in four patients whose tumors had a majority of large cleaved cells. These preliminary results, if confirmed in a larger series of patients, will provide additional evidence that there are consistent chromosome changes associated with specific subtypes of lymphoproliferative disorders analogous to the Ph1 chromosome in chronic myelogenous leukemia.
Variability of the homeostatic response to altered p50
Authors: Charache S , Achuff S , Winslow R , Adamson J , Chervenick P
Published: October 2019

Blood from carriers of hemoglobin Osler (Hb Osler) had almost the same oxygen affinity as that of carriers of Hb McKees Rocks (Hb MR) (P50 10- 11 mm Hg), but Hb concentrations were higher in male carriers of the former (21.6 versus 17.2 g/dl). Two carriers of each Hb were studied to compare their adaptations to altered oxygen affinity and their responses to phlebotomy. All four were healthy, and all excreted normal amounts of erythropoietin. Carriers of Hb MR had somewhat lower mixed venous pO2 than carriers of Hb Osler. There was no suggestion that phlebotomy impaired ability to exercise in either group of patients.
Chronic myelocytic leukemia (CML): failure to detect residual normal committed stem cells in vitro
Authors: Singer JW , Fialkow PJ , Steinmann L , Najfeld V , Stein SJ , Robinson WA
Published: October 2019

Granulocytic colonies grown in culture from marrow and peripheral blood from five patients with Ph1-positive CML and heterozygous at the G-6-PD locus were analyzed for G-6-PD in order to identify CFU-C that do not arise from the CML clone. The patients had both B and A enzymes in normal tissues, but their CML clones typed as B. Whereas about 50% of colonies from normal subjects heterozygous as the G-6-PD locus show type-A G-6-PD and 50% type B, only two of the 1308 colonies from the CML patients had type-A G-6-PD. These data provide little evidence for persistence of normal committed stem cells in CML, a finding in contrast to that made previously in polycythemia vera, another clonal stem cell myeloproliferative disorder.
Human IgA1 half-molecules: clinical and immunologic features in a patient with multiple myeloma
Authors: Sakurabayashi I , Kin K , Kawai T
Published: October 2019

Abnormal IgA1 half-molecules consisting of one heavy and one light chain were found in a patient (N.N.) with typical multiple myeloma. The serum and the urine of this patient contained both 7.0S and 3.9S IgA myeloma proteins. The IgA half-molecules (3.9S) were found to have a molecular weight of 59,000 daltons and were composed of one alpha1 chain of about 40,000 daltons and one light chain of 22,000 daltons. Furthermore, enzymatic degradation suggested that the alpha chain of the N.N. half-molecules had a large deletion in its Fc portion. We suggest that its heavy and light chains were probably bound noncovalently, since the interchains connecting the heavy and light chains of these IgA half-molecules were easily dissociated with 1% SDS and 8 M urea. Cytologic studies identified at least two types of myeloma cells, and it is possible that half-molecule IgA production might result from mutation among the myeloma cells producing whole- molecule IgA.
An increased Bohr effect in sickle cell anemia
Authors: Ueda Y , Nagel RL , Bookchin RM
Published: October 2019

Recent findings that hemoglobin S gelation and sickling are pH- dependent and also influence oxygen affinity suggested that the red cells containing this hemoglobin variant might show an abnormal Bohr effect. We therefore studied the effects of pH variation on the in vitro oxygen affinity of whole blood from persons with sickle cell anemia (SS) and normal donors (at 37 degrees C and constant carbon dioxide tension of 40 mm Hg). The Bohr effect in SS blood was greatly increased only between blood pH 7.4 and 7.2 (cell pH 7.2 and 7.0, a shift that strongly affects gelation), with delta log p50/deltapH= - 0.92 to -0.99 (normal = -0.42 to -0.46). Thus a drop in SS blood pH below 7.4 in tissue capillaries yields twice the normal decrease in oxygen affinity and a large release of oxygen from red cells, whose risk of sickling is high. Even mild transient acidosis would seem hazardous for patients with sickling disorders.
Hematologic and cytologic characterization of 8/21 translocation acute granulocytic leukemia
Authors: Trujillo JM , Cork A , Ahearn MJ , Youness EL , McCredie KB
Published: October 2019

Cytogenetic studies were performed in 546 patients with acute leukemia between 1968 and 1975. Two hundred thirty-four patients were aneuploid (42.9%), and 312 patients were diploid (57.1%). Among these, 32 patients were found to exhibit similar chromosomal alterations that appeared to involve specifically chromosomes 8 and 21. Banding studies in at least 15 of these patients confirmed the presence of a translocation between these two chromosomes. The cytogenetic findings were correlated with the hematologic and clinical data. It was found that each of these individuals had a typical picture of acute granulocytic leukemia with Auer rod-positive and peroxidase-positive cells. Ultrastructurally, the patients in this group also consistently demonstrated the presence of a nuclear bleb that has been positively associated with aneuploidy in acute leukemia. Clinically, they seemed to respond better to therapy than other adult patients with acute granulocytic leukemia. It is proposed that the 8/21 translocation acute leukemia represents a definite subgroup within the general category of acute granulocytic leukemia, with an incidence of approximately 7.3%.
Interference of levamisole with inhibition of E-rosette formation by Hodgkin's disease and systemic lupus erythematosus cytotoxic sera
Authors: Del Giacco GS , Tognella S , Leone AL , Locci F , Cornaglia P , Sangiuolo A , Grifoni V
Published: October 2019

A new system has been used to test the influence of levamisole on T- cell function. Evidence has been produced that prior exposure to the drug “protects” normal human peripheral blood lymphocytes from the inhibition that cytotoxic sera from patients with Hodgkin's disease and systemic lupus erythematosus exhibit on their E-rosette-forming capacity. Also, damage of this T-cell function already induced by Hodgkin's sera may be partially corrected.
Identification of megakaryocytes, macrophages, and eosinophils in colonies of human bone marrow containing neurtophilic granulocytes and erythroblasts
Authors: Fauser AA , Messner HA
Published: October 2019

Pluripotent hemopoietic progenitors in human bone marrow can be identified by their ability to give rise in culture to colonies that contain more than one lineage of hemopoietic differentiation. Growth of these mixed colonies is supported by media conditioned by leukocytes in the presence of phytohemagglutinin (PHA-LCM) and erythropoietin. They can be readily recognized by direct microscopic observation because of their composition of colorless cells and cells with a red appearance typical for hemoglobin. Seventy-three individual mixed colonies from 6 individuals were prepared by cytocentrifugation for further cytological examination. In addition to neutrophilic granulocytes and erythroblasts, megakaryocytes were present in 16 of the 73 colonies, as identified by positive reaction for acid phosphatase. Macrophages were found in 24 and eosinophils in 6 of the 73 colonies.
Protease activity in the human erythrocyte: localization to the cell membrane
Authors: Ballas SK , Burka ER
Published: October 2019

Proteolytic activity in the mature human erythrocyte was localized to the cell membrane. This activity, which could not be ascribed to contaminating leukocytes in the suspension, was totally absent from the soluble portion of the cell and was more active against hemoglobin subunits than against tetrameric hemoglobin molecules. Pulse-chase experiments confirmed that the membrane proteolytic activity was more active in degrading beta chains that alpha chains. The activity could be extracted from erythrocyte cell membranes, but not from membrane- free hemolysate, by exposure to 0.75-M KSCN. The activity of the protease was dependent on time and temperature and did not require ATP or an energy-generating system for activity.
Interrelationship of immunologic characteristics, proliferation pattern, and prednisone sensitivity in acute lymphoblastic leukemia of childhood
Authors: Bakkeren JA , de Vaan GA , Hillen HF
Published: October 2019

In children with acute lymphoblastic leukemia (ALL) the effect of prednisone therapy on the cell-cycle phase distribution of leukemic bone marrow cells was determined with pulse cytophotometry at the time of diagnosis. Also, the interrelationship with the presence of the sheep erythrocyte receptor as a marker for T cells was investigated. In 17 or 21 patients prednisone treatment caused a decrease in the percentage of cells in the S+G2+M phases. In 11 of 12 adult patients with ALL the same result was obtained. In a group of 31 children with ALL, 6 cases of T-cell ALL occurred in combination with intermediate or high WBC counts. The 3 patients with the highest E-rosetting percentages had rather high proportions of bone marrow cells in the S+G2+M phases. No correlation could be established between the WBC count or the E-rosetting percentage and the chance for complete remission.
Platelet alpha granules contain a growth factor for fibroblasts
Authors: Kaplan DR , Chao FC , Stiles CD , Antoniades HN , Scher CD
Published: October 2019

Platelets contain a polypeptide growth factor that stimulates the replication of normal connective tissue cells; this platelet-derived growth factor (PDGF) is released during the clotting process. Human platelets from normal volunteers were disrupted by nitrogen cavitation, and the subcellular organelles were fractionated by ultracentrifugation through a 30%--60% sucrose gradient. Electron microscopy revealed that fraction 7 (density 1.23 g/liter) contained the largest number of alpha granules. The specific activity of platelet fibrinogen, an alpha- granule marker, was also highest in this fraction. The subcellular fractions were assay for the presence of PDGF and for beta- thromboglobulin. PDGF was assayed quantitatively by the stimulation of DNA synthesis in confluent growth-arrested BALB/c-3T3 cells, whereas the concentration of beta-thromboglobulin was determined by radioimmunoassay. The highest concentrations of both PDGF and beta- thromboglobulin were found in the alpha-granule fraction. In contrast, beta-glucuronidase, a lysosomal enzyme, was more diffusely distributed and had its highest specific activity in fractions of lower density than those for PDGS, beta-thromboglobulin, or fibrinogen. The data demonstrate that the alpha granules of platelets provide a unique delivery system for PDGF, a polypeptide hormone with growth-promoting activity for connective tissue cells.
Nonfatal graft-versus-host disease occurring after transfusion with leukocytes and platelets obtained from normal donors
Authors: Cohen D , Weinstein H , Mihm M , Yankee R
Published: October 2019

There have been two previous reports of fatal graft-versus-host (GVH) disease following transfusion of leukocytes from normal donors for the supportive care of leukemic patients. We have reported a nonfatal case of GVH disease in a child with acute monoblastic leukemia who received leukocyte and platelet transfusions from normal donors. The dose of lymphocytes contained in both leukocyte and platelet concentrates obtained with a semicontinuous-flow cell separator is sufficient to cause GVH disease. Irradiation of all blood products administered to severely immunocompromised and myelosuppressed patients should beconsidered.