bluuebunny/crossref_metadata_2025_split huggingface dataset:
Influence of hydroxyurea on fetal hemoglobin production in vitro
Authors: Miller BA , Platt O , Hope S , Dover G , Nathan DG 
Published: October 2019

Abstract
Cytotoxic drugs increase circulating fetal hemoglobin levels. We examined the mechanism by measuring the fetal hemoglobin produced per BFU-E-derived erythroblast following hydroxyurea treatment in vivo and in vitro. Treatment of four sickle cell patients increased the percentage of circulating F reticulocytes. The frequencies of bone marrow or peripheral blood BFU-E or CFU-E-derived colonies and their fetal hemoglobin content were unaffected. In all cases, the number of erythroid cells/progenitor-derived colony increased. To explore further the effect of hydroxyurea on fetal hemoglobin production, we added 50 mumol/L hydroxyurea to cultures of peripheral blood BFU-E-derived erythroblasts on 1 of 9 days (day 5 through 13) to nine samples. These BFU-E were derived from the peripheral blood of normal donors, sickle trait donors, and sickle cell anemia patients and from the bone marrows of monkeys. This concentration of hydroxyurea was selected so that the frequency of BFU-E and their size was moderately decreased. Addition of hydroxyurea to these progenitor-derived erythroid cells had no effect on fetal hemoglobin content per cell. Neither did transient exposure of progenitors to hydroxyurea prior to culture in nontoxic concentrations (0 to 500 mumol/L) result in a significant increase in fetal hemoglobin content in progenitor-derived erythroblasts. These data suggest that hydroxyurea does not directly alter the HbF program expressed by progenitor-derived erythroid cells. Instead, it enhances hemoglobin F content secondarily, possibly by inducing alterations in erythropoiesis.
Delta glycophorin (glycophorin B) gene deletion in two individuals homozygous for the S--s--U-- blood group phenotype
Authors: Huang CH , Johe K , Moulds JJ , Siebert PD , Fukuda M , Blumenfeld OO 
Published: October 2019

Abstract
Blood cells from two unrelated individuals whose erythrocytes exhibit respectively N S-s-U- and MN S-s-U- blood group phenotypes were examined by immunoblotting, periodic acid-Schiff (PAS) staining, and Southern blotting. Protein bands characteristic of delta glycophorin (glycophorin B) were absent from the immunoblots of whole erythrocyte lysates when probed with polyclonal glycophorin antisera and from isolated erythrocyte membranes stained with PAS reagents. Genomic DNA from the two individuals' leukocytes was digested with a panel of restriction enzymes and probed with alpha M glycophorin cDNA obtained from human K562 leukemic cell line. The EcoRI, PstI, and KpnI restriction site patterns were identical to those of S+s+U+ controls in fragment numbers and relative size but differed from controls in band intensities. Restriction mapping with HindIII, PvuII, SacI, MspI, and BamHI revealed that S-s-U- individuals lack fragments that are reproducibly observed in S+s+U+ controls, and most likely encode delta glycophorin. Using truncated 5′ and 3′ cDNA segments as probes and comparing, in control individuals, hybridization intensities of fragments with amino acid sequence homologies, we have inferred the assignment of restriction fragments to the alpha and delta glycophorin genes. Our results suggest that the absence of delta glycophorin in the two S-s-U- individuals is a result of deletion of the entire delta glycophorin gene. This is the first report of a glycophorin gene deletion.
Homing receptors on human and rodent lymphocytes--evidence for a conserved carbohydrate-binding specificity
Authors: Stoolman LM , Yednock TA , Rosen SD 
Published: October 2019

Abstract
Lymphocyte recirculation begins with the attachment of circulating cells to the structurally distinctive postcapillary venules of lymphoid organs termed high-endothelial venules (HEVs). In both rodents and humans, the attachment of lymphocytes to the HEVs of peripheral lymph nodes (PNs) on the one hand and gut-associated lymphoid tissues (GALTs) on the other appears to involve discrete adhesive structures on the surfaces of the interacting cells. In rodents, we previously showed that a carbohydrate-binding receptor at the lymphocyte surface participates in the attachment to the HEV of peripheral nodes. The studies reported herein document the involvement of a similar receptor in the selective attachment of human peripheral blood lymphocytes to the HEVs of PNs. We argue that the close functional relationship between the human and rodent receptors indicates that this component of the adhesive interaction has been conserved through evolution.
Spontaneous production of interleukin 1 activity by chronic lymphocytic leukemic cells
Authors: Uggla C , Aguilar-Santelises M , Rosen A , Mellstedt H , Jondal M 
Published: October 2019

Abstract
Chronic lymphocytic leukemic B cells (B-CLL) were found to produce an IL 1-like growth factor spontaneously in vitro for mouse thymocytes. This factor was comitogenic with concanavalin A (Con A) and nonmitogenic combinations of phorbol ester and calcium ionophore but not with phyto-hemagglutinin (PHA). Growth factor production was dose- related to the number of in vitro cultured cells and detectable at 6 hours using high cell concentrations. A small number of admixed normal T cells was not important for factor production. No growth of autologous B-CLL or allogeneic thymocytes was induced by the factor. A chromatographic high-performance liquid chromatography (HPLC) analysis and inhibition experiments with a polyclonal rabbit anti interleukin 1 (IL 1) antiserum indicated that the B-CLL-derived growth factor belonged to the IL 1 family. This was supported by the direct demonstration of IL 1 beta in supernatants from B-CLL by radioimmunoassay. Possible biologic implications for B-CLL-derived IL 1 are discussed in relation to tumor cell growth in different clinical stages.
Dermatan sulphate induces plasminogen activator release in the perfused rat hindquarters
Authors: Abbadini M , Zhu GJ , Maggi A , Pangrazzi J , Donati MB , Mussoni L 
Published: October 2019

Abstract
Heparin or heparin-like substances have been described to induce the release of plasminogen activator (PA) activity in different animal perfusion models. In this paper we report that Dermatan Sulphate (DS) is able to induce PA activity release in the perfused rat hindquarters. Perfusion of different doses of DS (0.1 to 0.8 mg/mL) stimulates a release of PA activity that is maximum after the initial two minutes of perfusion. The amount of PA activity released rises progressively within a certain concentration range of DS (0.1 to 0.4 mg/mL) and declines thereafter (0.6 to 0.8 mg/mL). The type of PA activity increased during DS perfusion was characterized by SDS-PAGE and fibrin autography as tissue-type PA (t-PA) on the basis of its mol wt (67,000 d) and inhibition by a specific anti t-PA antiserum. This effect might be considered as potentially contributing to the antithrombotic activity of DS, at least at the local level.
Deoxygenation inhibits the volume-stimulated, Cl(-)-dependent K+ efflux in SS and young AA cells: a cytosolic Mg2+ modulation
Authors: Canessa M , Fabry ME , Nagel RL 
Published: October 2019

Abstract
We recently reported that the Cl(-)-dependent K+ (K:Cl) efflux, which can be stimulated by cell swelling in the presence of inhibitors of the Na+ pump (ouabain) and of the Na-K-Cl cotransport (bumetanide), is highly active in young AA and SS RBCs. We report here that deoxygenation inhibits volume-stimulated K:Cl efflux in SS and reticulocyte-enriched density-separated SS and AA RBCs. In SS whole blood, the K:Cl efflux stimulated by hypotonic (220 mOsm) swelling was reduced from 9.2 +/- 2 (mean +/- SE) in oxygen to 2.7 +/- 1.9 (mmol/L cell/h = flux units = FU) (n = 4) under deoxygenated conditions (P less than .005). Deoxygenation also decreased the acid pH-stimulated K:Cl efflux from 5.9 +/- 1.5 to 3.7 +/- 1.1 FU (n = 3) (P less than .025) but did not inhibit NEM-stimulated K:Cl transport. The effect of deoxygenation on density-separated SS cells is similar: When fraction SS2 (reversible discocytes) is deoxygenated under hypotonic conditions, the K:Cl efflux is reduced by 50%. In reticulocyte-enriched AA cells obtained from anemic patients, deoxygenation under hypotonic conditions also reduces K+ efflux by 50%. In SS cells only, deoxygenation under isotonic conditions results in an increased Cl(-)-independent K+ efflux. Because ionized Mg2+ in the cytosol increases during deoxygenation, we investigated the effect of external and internal Mg2+ on the volume-stimulated K:Cl efflux. Removal of external Mg2+ did not influence the rate of transport in oxygenated cells. When internal Mg2+ was clamped at 0.15 mmol/L with A23187 and EDTA at ionized cytosolic Ca2+ = O, however, the inhibitory effect of deoxygenation on the K:Cl efflux was eliminated. We conclude that deoxygenation inhibits the volume-stimulated, Cl(-)-dependent K+ efflux in AA and SS young red cells by concomitantly increasing ionized cytosolic Mg2+.
Cytogenetic studies and in vitro colony growth in patients with mastocytosis
Authors: Swolin B , Rodjer S , Roupe G 
Published: October 2019

Abstract
Cytogenetic analysis of bone marrow cells and in vitro growth for bone marrow granulocytic-macrophage stem cells have been performed in 13 patients with mastocytosis, six with systemic mastocytosis, and seven with urticaria pigmentosa. Clones with chromosome abnormalities were found in five patients. The number of clusters and/or colonies after seven days in culture was increased in seven patients, compared with the growth in a control group. Three patients with chromosome abnormalities showed an abnormal growth pattern, yet exhibited normal peripheral blood values. Two patients with systemic mastocytosis had clones with chromosome abnormalities and some abnormal hematological values. The proportion of patients with chromosome abnormalities and an abnormal growth pattern was higher among these patients with mastocytosis than in healthy control subjects. These results may be of interest when discussing the origin of mast cell disorders and indicate an association with the myeloproliferative disorders.
Host origin of the human hematopoietic microenvironment following allogeneic bone marrow transplantation
Authors: Laver J , Jhanwar SC , O'Reilly RJ , Castro-Malaspina H 
Published: October 2019

Abstract
The origin of marrow stromal cells post allogeneic bone marrow transplantation (BMT) was studied. Two groups of patients receiving HLA- identical marrow grafts from sex mismatched siblings were included in the study: the first group (eight patients) received conventional marrow grafts and the second group (ten patients) received stromal cell and T cell depleted grafts. All patients showed hematopoietic engraftment with donor cells. Marrow aspirates obtained from these patients were used to establish stromal layers in long-term marrow cultures (LTMC) for 4 to 6 weeks. In both groups, karyotype analysis of nonhematopoietic cultured stromal cells showed host origin even as late as day 760 posttransplantation. Immunofluorescence methods using monoclonal antibodies against components of fibroblasts, macrophages, and endothelial cells, showed that the composition of stromal layers was similar to those obtained from normal controls. Our data indicate that marrow stromal progenitors capable of proliferation are nontransplantable and do not originate from a hematopoietic-stromal common progenitor.
Identification of high-affinity (Kd 0.35 mumol/L) and low-affinity (Kd 7.9 mumol/L) platelet binding sites for ADP and competition by ADP analogues
Authors: Jefferson JR , Harmon JT , Jamieson GA 
Published: October 2019

Abstract
Steady-state binding of ADP to blood platelets and isolated membranes has not previously been obtained because of complications arising from metabolism of the ligand and dilution due to its secretion from storage granules. In the present studies, competition binding isotherms (n = 9) using paraformaldehyde-fixed platelets showed that [2–3 H]ADP bound to two sites with a small amount (approximately 5% of total) of nonspecific binding: 410,000 +/- 40,000 sites of low affinity (Kd 7.9 +/- 2.0 mumol/L) and 160,000 +/- 20,000 sites of high affinity (Kd 0.35 +/- 0.04 mumol/L) corresponding to the ADP concentration required for activation in fresh platelets (0.1–0.5 mumol/L). All agonists and antagonists examined were able to compete with ADP at the high-affinity site. The strong platelet agonists 2-methylthio ADP and 2-(3- aminopropylthio)ADP competed with ADP at the high-affinity site with dissociation constant values of 7 mumol/L and 200 mumol/L, respectively. The partial agonist 2′,3′-dialdehyde ADP and the weak agonist GDP also competed at the high-affinity site with Kd values of 5 mumol/L and 49 mumol/L, respectively. The sequence of binding affinities of other adenine nucleotides at the high-affinity site corresponded to their relative activities as known antagonists of platelet activation by ADP; namely, ADP(Kd 0.35 mumol/L) approximately equal to ATP (Kd 0.45 mumol/L) much greater than AMP (Kd 360 mumol/L). Adenosine and 2-chloroadenosine did not compete with ADP. ADP binding to the high-affinity site was inhibited by p-mercuribenzene sulfonate (Ki 250 mumol/L) but only very weakly by 5′-p- fluorosulfonylbenzoyladenosine (Ki 1 mmol/L). All the above nucleotides also competed with ADP at the low-affinity sites but, because of the high concentrations of competing nucleotide required, dissociation constants at this site were obtained only for ATP (21 mumol/L), 2-MeS ADP (200 mumol/L) and 2′,3′-dialdehyde ADP (270 mumol/L). 8-Bromo ADP competed strongly with ADP at the high-affinity site (Kd 0.40 mumol/L) but weakly if at all at the low-affinity site. 8-Bromo ADP inhibited platelet activation induced by ADP (EC50 approximately 100 mumol/L) but not by collagen, thrombin, or ionophore A23187.(ABSTRACT TRUNCATED AT 400 WORDS).
Platelet-activating factor induces protein kinase activity in the particulate fraction of human neutrophils
Authors: Gay JC , Stitt ES 
Published: October 2019

Abstract
Platelet-activating factor (PAF) is a proinflammatory lipid that has both platelet- and phagocyte-stimulating properties. Because several known activators of calcium-, phospholipid-dependent protein kinase (protein kinase c, PKC) also stimulate neutrophil responses and because neutrophil stimuli such as phorbol diesters and the chemotactic peptide f-Met-Leu-Phe are reported to increase protein kinase activity in neutrophil (PMN) particulate fractions, we investigated the effect of PAF on neutrophil protein kinase activities. In neutrophils exposed to 10(-6) mol/L PAF, cytosolic PKC activity was 521 +/- 38 pmol 32P/10(7) PMN/min (mean +/- SEM), which was not significantly lower than cystolic activity in buffer-treated controls (558 +/- 32 pmol 32P/10(7) PMN/min, n = 14). PAF-exposed cells exhibited a concomitant rise in protein kinase activity associated with the particulate fraction with 53 +/- 4 pmol 32P/10(7) PMN/min compared with 32 +/- 2 pmol in control cells (n = 14). Particulate protein kinase activity was independent of the presence of calcium and phospholipid in the assay medium. The specific PKC inhibitor H-7 inhibited particulate protein kinase activity, however, which suggested that the enzyme activity assayed in this fraction may be PKC in a constitutively activated form. The increase in particulate protein kinase activity induced by PAF required the presence of cytochalasin B, was detectable within 5 seconds of exposure to PAF, and was not reversed by washing the cells free of extracellular PAF after initial exposure. Although PAF did not have a direct effect on PKC activity from cytosolic fractions from resting cells, the increase in particulate protein kinase activity induced by PAF was inhibited when the cells were first depleted of calcium by incubation with Quin 2. These results suggest that PAF induces an increase in particulate protein kinase activity in neutrophils by a calcium- dependent mechanism and that the induction of membrane-associated protein kinase activity may be involved in neutrophil-stimulating actions such as superoxide production, which occur at higher concentrations of PAF.
Evaluation of drugs for elimination of leukemic cells from the bone marrow of patients with acute leukemia
Authors: Auber ML , Horwitz LJ , Blaauw A , Khorana S , Tucker S , Woods T , Warmuth M , Dicke KA , McCredie KB , Spitzer G 
Published: October 2019

Abstract
Relatively nonmyelotoxic drugs and drug combinations were investigated for their ability to eliminate malignant cells from human bone marrow. In vitro 90% inhibitory concentration (IC90) doses were established on granulocyte macrophage colony-forming units (GM-CFU) in culture of bone marrow by using the GM-CFU assay for the following drugs: 4- hydroperoxycyclophosphamide (4-HC), Adriamycin, L-asparaginase, bleomycin, hydrocortisone, VP-16, spirogermanium, Taxol, and vincristine. The leukemic cell kill efficiency of these drugs at IC90 doses was compared with that of 4-HC on acute lymphoid leukemia (ALL) cell lines by using the limiting-dilution assay. Under these conditions, no single drug was superior to 4-HC. To increase the in vitro effect in leukemic cell kill, combinations of vincristine with hydrocortisone, Adriamycin, VP-16, and 4-HC were investigated. Vincristine at 1 to 5 micrograms/mL increased the marrow cytotoxicity of hydrocortisone, Adriamycin, and VP-16, but it was protective (subadditive) with 4-HC. Vincristine and 4-HC in combination was additive to supraadditive on ALL cell lines, increased the leukemic cell kill by one to two logs above 4-HC alone at IC90 doses (P less than .05), and was not affected by the addition of excess marrow cells. The recommended doses for chemopurging in clinical studies are vincristine, 1 to 5 micrograms/mL, plus 4-HC, 5 micrograms/mL.
Phenotypic and molecular heterogeneity in Philadelphia chromosome- positive acute leukemia
Authors: Hirsch-Ginsberg C , Childs C , Chang KS , Beran M , Cork A , Reuben J , Freireich EJ , Chang LC , Bollum FJ , Trujillo J 
Published: October 2019

Abstract
Philadelphia chromosome-positive (Ph1) acute leukemia is a heterogeneous subset of acute leukemia with a poor prognosis. We studied five patients to determine the potential for phenotypic and molecular heterogeneity. Cellular characterization studies included light myeloperoxidase (L-MPO), terminal deoxynucleotidyl transferase (TdT), ultrastructural MPO (U-MPO), and immunophenotyping by flow cytometry using T11, T3, T4, T8, Leu 1, B1, Leu 12, HLA-DR (la), CALLA (J5), OKM1, My4, My7, My8, My9, and My10. DNA was analyzed for rearrangements of the breakpoint cluster region (bcr), immunoglobulin heavy chain, joining region (JH), immunoglobulin kappa light chain constant region (C kappa), and T cell receptor (TcR beta). RNA dot blots were hybridized by using molecular probes for MPO and TdT. We found that four of five cases were acute mixed-lineage leukemia (AMLL). One patient had acute unclassifiable leukemia. Of the four patients classified as having AMLL, three showed myeloid and lymphoid features, with one patient showing myeloid, T cell, and B cell features. The last case showed T cell and B cell features only. In one patient MPO/RNA was positive in spite of insufficient L-MPO or U-MPO to diagnose acute myelogenous leukemia (AML), thereby suggesting significant MPO gene expression before the production of sufficient MPO protein to meet the French-American-British criteria for AML. Three of the five patients showed rearrangement of bcr (cases 1, 2, and 5). Studies of these five patients support the concepts of molecular and phenotypic heterogeneity in Ph1 acute leukemia, demonstrate a high incidence of AMLL in this subset of acute leukemia, and support the use of lineage-associated molecular probes to define lineage at an earlier stage than previously possible.
Sodium butyrate and a T lymphocyte cell line-derived differentiation factor induce basophilic differentiation of the human promyelocytic leukemia cell line HL-60
Authors: Hutt-Taylor SR , Harnish D , Richardson M , Ishizaka T , Denburg JA 
Published: October 2019

Abstract
Sodium butyrate induces basophilic differentiation of HL-60 promyelocytic leukemia cells that have been previously passaged in alkaline medium. A factor present in Mo conditioned medium (Mo-CM) acts synergistically with sodium butyrate to promote basophilic maturation in a dose-dependent fashion. The induced HL-60 cells exhibit nuclei at various stages of maturity and cytoplasmic granules staining azurophilic with May-Grunwald-Giemsa and metachromatically with toluidine blue. The histamine content of induced HL-60 cells is 50 ng/10(6) cells with sodium butyrate alone or 190 ng/10(6) cells with butyrate in combination with Mo-CM. Induced cells release histamine in response to anti-IgE and have receptors for the Fc portion of human IgE. The basophilic cell-differentiating activity present in Mo-CM appears to be distinct from several other cytokines including recombinant human interleukin-1 alpha, interleukin-2, interferon-gamma, interferon-alpha, murine interleukin-3, erythroid-potentiating activity, and purified human granulocyte/macrophage colony-stimulating factor. This is the first demonstration of a cell line that is capable of differentiation along the basophil lineage and could provide a useful model for examining biochemical and molecular events associated with basophil differentiation.
Expression of tissue factor and factor VIIa/tissue factor inhibitor activity in endotoxin or phorbol ester stimulated U937 monocyte-like cells
Authors: Rana SV , Reimers HJ , Pathikonda MS , Bajaj SP 
Published: October 2019

Abstract
Previously, unstimulated cells of the human monocytic tumor cell line U937 have been shown to possess a negligible cell-surface tissue factor (TF) activity, and to secrete a small amount of factor VIIa/tissue factor (VIIa/TF) inhibitor activity. On stimulation with endotoxin or with phorbol myristate acetate (PMA), TF of these cells is known to be increased approximately fourfold. In this report, we demonstrate that VIIa/TF inhibitor is also increased on stimulation of U937 cells with endotoxin (approximately equal to threefold) or with PMA (approximately equal to 20-fold). Notably, the secretion of the inhibitor persisted after the cell surface TF had started to decline. Further, when serum- free media from PMA stimulated cells was electrophoresed on a sodium dodecyl sulfate (SDS) gel, we eluted two inhibitor activity peaks corresponding to Mr approximately equal to 47,000 and Mr approximately equal to 36,000. The molecular weights of these peaks are similar to those obtained earlier from human plasma for this inhibitor(s).
Opposite effects of recombinant interferon-alpha A and deoxycoformycin on adenosine deaminase activity in the Daudi B lymphoblastoid cell line
Authors: Faltynek CR 
Published: October 2019

Abstract
Interferon-alpha and the adenosine deaminase (ADA) inhibitor deoxycoformycin (dCF) have each been shown to be efficacious in the treatment of some lymphoid malignancies and to have potent antiproliferative activities in vitro. This study examined whether dCF and recombinant interferon-alpha A (rIFN-alpha A) were additive, synergistic, or antagonistic in their effects on the cultured B lymphoblastoid cell line Daudi. Treatment of Daudi cells for three to four days with doses of rIFN-alpha A that were growth inhibitory was unexpectedly found to increase the level of ADA activity per cell two- to threefold and therefore to prevent the inhibition of ADA by limiting concentrations of dCF. However, the opposite effects of dCF and rIFN- alpha A on ADA activity did not lead to antagonistic effects on growth inhibition. The higher concentrations of dCF (with deoxyadenosine) necessary for appreciable growth inhibition could inhibit the increased ADA activity in rIFN-alpha A-treated cells, thus resulting in additive antiproliferative effects.