bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fractionation of subsets of BFU-E from normal human bone marrow: responsiveness to erythropoietin, human placental-conditioned medium, or granulocyte-macrophage colony-stimulating factor
Authors: Kannourakis G , Johnson GR 
Published: October 2019

Abstract
Normal human bone marrow mononuclear cells were fractionated by differential adherence, immunomagnetic separation, and fluorescence- activated cell sorting (FACS). The resultant fractionated cells were cultured in semisolid medium to monitor the presence of BFU-E, Mix-CFC, and nonerythroid CFC. Two populations of cells were recovered on the basis of binding by the monoclonal antibody (MoAb) RFB-1. One of these populations contained BFU-E that were stimulated only by erythropoietin (Epo), whereas the second population contained BFU-E responsive to Epo, Epo and recombinant human granulocyte-macrophage colony-stimulating factor (rHGM-CSF), or Epo and human placental-conditioned medium (HPCM). Prior enrichment of clonogenic cells by removal of adherent and Leu-M3+ve, Leu-4+ve, Leu-7+ve, B1+ve, WEMG1+ve, and Glycophorin A+ve cells, followed by FACS fractionation on the basis of RFB-1 binding, consistently resulted in recoveries of BFU-E, Mix-CFC, and nonerythroid CFC of greater than 100% (up to 800%). These procedures also resulted in enrichment of up to 200-fold and frequencies of 1:6 for BFU-E, 1:5 for CFC, and 1:130 for Mix-CFC.
A single-base change at position -175 in the 5'-flanking region of the G gamma-globin gene from a black with G gamma-beta+ HPFH
Authors: Surrey S , Delgrosso K , Malladi P , Schwartz E 
Published: October 2019

Abstract
Hereditary persistence of fetal hemoglobin (HPFH) is a human hemoglobinopathy characterized by the continued expression of fetal globins during adult life. Both deletional and nondeletional forms have been described. A number of single-base changes in the immediate 5′- flanking region of the fetal G gamma and A gamma have been reported associated with nondeletional forms of HPFH. We now present the nucleotide sequence of a G gamma-globin gene from an American black with G gamma-beta + HPFH. The immediate 5′-flanking region of this G gamma gene has a T-to-C change at -175, C at -158, and a normal C at - 202. Additional changes were found in IVS2 and in the immediate 3′- flanking region, some of which may represent gene-conversion events. The sequence change at -175 probably represents a second mutation associated with the G gamma-beta + HPFH phenotype in blacks. This base change alters an octamer sequence known to be of importance in the normal expression of several other genes.
Sardinian G gamma-HPFH: a T----C substitution in a conserved "octamer" sequence in the G gamma-globin promoter
Authors: Ottolenghi S , Nicolis S , Taramelli R , Malgaretti N , Mantovani R , Comi P , Giglioni B , Longinotti M , Dore F , Oggiano L 
Published: October 2019

Abstract
A survey of hemoglobinopathies in Northern Sardinia allowed the identification of two subjects heterozygous for a new type of G gamma hereditary persistence of fetal hemoglobin (HPFH). The G gamma-globin gene from the HPFH chromosome shows the presence of a T----C substitution 175 nucleotides upstream of the CAP site, adding a new example of single-point mutations occurring in the promoter region of the gamma-globin genes and linked to HPFH phenotypes. In this case the mutation affects the 3′ end nucleotide of a conserved octamer sequence known to be present in other regulatory elements of several genes.
Defective T cell-mediated, isotype-specific immunoglobulin regulation in B cell chronic lymphocytic leukemia
Authors: Moore JS , Prystowsky MB , Hoover RG , Besa EC , Nowell PC 
Published: October 2019

Abstract
The consistent occurrence of T cell abnormalities in patients with B cell chronic lymphocytic leukemia (B-CLL) suggest that the non- neoplastic host T cells may be involved in the pathogenesis of this B cell neoplasm. Because potential defects of immunoglobulin regulation are evident in B-CLL patients, we investigated one aspect of this by studying the T cell-mediated immunoglobulin isotype-specific immunoregulatory circuit in B-CLL. The existence of class-specific immunoglobulin regulatory mechanisms mediated by Fc receptor-bearing T cells (FcR + T) through soluble immunoglobulin binding factors (IgBFs) has been well established in many experimental systems. IgBFs can both suppress and enhance B cell activity in an isotype-specific manner. We investigated the apparently abnormal IgA regulation in a B-CLL patient (CLL249) whose B cells secrete primarily IgA in vitro. Enumeration of FcR + T cells showed a disproportionate increase in IgA FcR + T cells in the peripheral blood of this patient. Our studies showed that the neoplastic B cells were not intrinsically unresponsive to the suppressing component of IgABF produced from normal T cells, but rather the IgABF produced by the CLL249 host T cells was defective. CLL249 IgABF was unable to suppress IgA secretion by host or normal B cells and enhanced the in vitro proliferation of the host B cells. Size fractionation of both normal and CLL249 IgABF by gel-filtration high- performance liquid chromatography (HPLC) demonstrated differences in the ultraviolet-absorbing components of IgABF obtained from normal T cells v that from our patient with defective IgA regulation. Such T cell dysfunction may not be restricted to IgA regulation, since we have found similar expansion of isotype-specific FcR + T cells associated with expansion of the corresponding B cell clone in other patients with B-CLL. These data suggest that this T cell-mediated regulatory circuit could be significantly involved in the pathogenesis of B-CLL.
Antigenic phenotype and functional characterization of human tonsil B cells
Authors: Gadol N , Peacock MA , Ault KA 
Published: October 2019

Abstract
Human tonsil cells were labeled with anti-leu-16, a monoclonal antibody (MoAb) that recognizes the CD20 antigen and that is specific for B cells. Two populations of B cells were identified by flow cytometry on the basis of antigen density. One labeled brightly with anti-Leu-16, and the other labeled at a level comparable to blood B cells. These two populations were characterized with a panel of MoAbs in two- and three- color flow cytometric studies and appeared to correspond to germinal- center and mantle-zone B cells. The pattern of staining of anti-Leu-16 on sections of frozen tonsil supported this characterization. Anti-Leu- 16 labeled germinal center cells more intensely than mantle zone cells and stained a few scattered B cells in the interfollicular zone. The ability of each Leu-16+ population to secrete IgG and IgM in response to mitogens was measured in a particle immunofluorescence assay. Dim Leu-16+ B cells (small, resting B cells and a subpopulation of preactivated cells) secreted IgG and IgM in response to pokeweed mitogen (PWM) but only IgG in response to B cell growth factor (BCGF). Bright Leu-16+ B cells (small to large activated cells and possibly memory cells) did not respond to PWM but secreted IgG in response to BCGF. The functional responses of dim Leu-16+ and bright Leu-16+ B cells were consistent with their identification as mantle-zone and germinal-center B cells. Phenotypic identification and functional studies of mantle-zone and germinal-center B cells may help clarify the differentiation pathway within the germinal center.
Immunohistochemical characterization of two new monoclonal antibodies (LN-4, LN-5) reactive with human macrophage subsets and derived malignancies in B5-fixed, paraffin-embedded tissues
Authors: Bhoopat L , Turner RR , Stathopoulos E , Meyer PR , Taylor CR , Marder RJ , Epstein AL 
Published: October 2019

Abstract
Two monoclonal antibodies (LN-4, LN-5) reactive to human macrophages in B5 formalin-fixed, paraffin-embedded tissue sections have been produced by using deparaffinized cell extracts of peripheral blood mononuclear cells. Both monoclonal antibodies were initially identified on paraffin- embedded sections of hyperplastic lymph nodes by using the immunoperoxidase staining procedure. Specificity screens on normal human tissues show that LN-4 and LN-5 stain the cytoplasm of macrophages and histiocytes in hematopoietic organs including Kupffer's cells of the liver and Langerhans' cells of the skin. LN-4 also showed strong positivity with acini of the stomach, whereas LN-5 was positive with mantle zone B lymphocytes of the lymph node and spleen, spermatogonia, and chief cells of the stomach. Both antibodies were strongly reactive with cases of true histiocytic lymphoma but, except for infiltrating macrophages, were entirely negative in Hodgkin's disease and non-Hodgkin's lymphomas. In all cases of nodular sclerosis Hodgkin's disease, LN-4 was positive in macrophagelike cells present in the collagen bands surrounding the Hodgkin's lesions. Both monoclonal antibodies were also positive in macrophages and histiocytes present in a variety of benign lymphoid lesions including persistent generalized lymphadenopathy, Gaucher's disease, sinus histiocytosis, and dermatopathic lymphadenopathy. Because of their specificity for human macrophages, and their ability to stain B5-fixed, paraffin-embedded tissues, LN-4 and LN-5 are important new reagents for the diagnosis and classification of malignant and benign histiocytic lesions.
Fibrinolysis during liver transplantation in humans: role of tissue- type plasminogen activator
Authors: Dzik WH , Arkin CF , Jenkins RL , Stump DC 
Published: October 2019

Abstract
Human liver transplantation is frequently associated with a coagulopathy and bleeding diathesis developing during the anhepatic phase of surgery. The hemostatic defect has been attributed in part to accelerated fibrinolysis. In this study we evaluated changes in specific blood fibrinolytic parameters occurring in eight adult patients undergoing first-time orthotopic liver transplantation. Five of the eight patients experienced moderate to severe systemic fibrinolysis as reflected by alpha 2-antiplasmin consumption and fibrinogen degradation with the concomitant appearance of fibrin(ogen) degradation products. In association with these changes, an increase in tissue-type plasminogen activator (t-PA) activity and t-PA antigen levels was also observed. Fibrinolysis was most pronounced during the anhepatic phase of surgery and decreased after revascularization of the grafted liver. Three additional patients who underwent the same procedure manifested much less evidence of systemic fibrinolytic activation and had minimal elevation of t-PA antigen levels or activity. Urokinase-type plasminogen activator levels, although elevated in three patients, were disassociated from increased t-PA levels and concomitant systemic fibrinolysis. The operative course of those patients developing t-PA-associated fibrinolysis was characterized by shock, acidosis, generalized bleeding, and a need for substantially greater blood product support during surgery. These findings suggest that the observed fibrinolytic defect is related to increased circulating plasma levels of t-PA, presumably resulting from a combination of increased intravascular release and decreased hepatic clearance of t-PA. These observations may have implications for intraoperative therapy for the transplant-related coagulopathy and its associated bleeding.
Gamma gene promoter and enhancer structure in Seattle variant of hereditary persistence of fetal hemoglobin
Authors: Gelinas RE , Rixon M , Magis W , Stamatoyannopoulos G 
Published: October 2019

Abstract
A variant of hereditary persistence of fetal hemoglobin (HPFH), first described in a patient from Seattle, was studied by structural analysis of the gamma-globin genes. A family study suggested that the determinant for this form of HPFH, in which the HbF contains both G gamma- and A gamma-globin chains, segregated with the beta S gene. No deletions or other abnormalities were detected in the fetal to adult globin gene region by genomic mapping studies. All four gamma-globin genes were isolated from a cosmid library, and allelic pairs of gamma- globin genes were distinguished by linkage to either the beta S- or beta A-globin gene. Nucleotide sequence analysis of the four gamma- globin gene promoters revealed a total of three discrepancies compared with a reference sequence, but these were judged unlikely to be the underlying determinants. Sequence analysis of the enhancer region located 3′ to the A gamma-globin gene from the putative HPFH chromosome revealed three base substitutions, whereas this region was normal in the A gamma-globin gene linked to the beta A gene. These data raise the possibility that an alteration of enhancer function rather than promoter function could be the basis for this condition.
Discordant effect of interferon on natural killer activity and tumor cell sensitivity to lysis in hairy cell leukemia
Authors: Lahat N , Aghai E , Kotler A , Kinarty A , Sobel E , Gruener N , Froom P 
Published: October 2019

Abstract
We studied the action of alpha-interferon (IFN) and interleukin-2 (IL- 2) on natural killer (NK)-rich fractions and autologous tumor cells from two patients with hairy cell leukemia (HCL). The addition of IFN or IL-2 to the NK-rich fractions resulted in a significant increase in NK activity against the autologous tumor cells. This stimulatory effect was blocked if the target hairy cells (HCs) were preincubated with either IFN or IL-2. Pretreatment of the HCs with anti-Tac antibody entirely prevented the blocking effect of IL-2 and partially the blocking effect of IFN. One patient was treated with recombinant alpha c-IFN. After 2 months there was a dramatic reduction in the number of HCs in the peripheral blood coincident with the loss of the protection effect of IFN against NK lysis of the patient's HCs. NK activity against autologous tumor cells correlated poorly with that against the K562 cell line. We conclude that there is a discordant effect of IFN and IL-2 on NK activity and HC sensitivity to lysis. The Tac receptor appears to play a role in this sensitivity. Caution should be exercised in extrapolating the effects of NK activity against K562 cells to those on HC targets.
Independent prognostic significance of a nuclear proliferation antigen in diffuse large cell lymphomas as determined by the monoclonal antibody Ki-67
Authors: Grogan TM , Lippman SM , Spier CM , Slymen DJ , Rybski JA , Rangel CS , Richter LC , Miller TP 
Published: October 2019

Abstract
To assess the prognostic significance of the growth fraction in diffuse large cell lymphoma (DLCL), we studied 105 DLCL patients with the monoclonal antibody Ki-67 applied to frozen tissue sections. Ki-67 detects a nuclear antigen associated with cell proliferation not found in resting cells. Ki-67 findings and other clinical prognostic factors were correlated with outcome using univariate and multivariate analyses in the proportional hazards model. High proliferative activity, defined as nuclear Ki-67 expression in greater than 60% of malignant cells (Ki- 67 greater than 60), was found to be a strong predictor of poor survival among these patients (P = .003, log-rank). The 19 patients with Ki-67 greater than 60% had a median survival of 8 months compared with a median survival of 39 months for the 86 patients with Ki-67 less than or equal to 60%. Examination of pretreatment clinical variables indicated the patient groups were similar with regard to age, sex, stage, B symptoms, tumor bulk, and lactate dehydrogenase (LDH). Both patient groups received comparable curative intent therapy and showed comparable complete response rate precluding treatment differences as modifying outcome. Multivariate analysis indicated Ki-67 is an independent predictor of survival (multivariate P = .006). Further statistical analysis using only B-cell DLCL patients treated with CHOP (63 patients) indicated that Ki-67 greater than 60 retained strong prediction of poor outcome (P = .002, log-rank) among this homogeneous group. We conclude that high proliferative activity (Ki-67 greater than 60) is an independent factor allowing laboratory prediction of probable poor outcome of DLCL.
Selective generation of erythroid burst-promoting activity by recombinant interleukin 2-stimulated human T lymphocytes and natural killer cells
Authors: Skettino S , Phillips J , Lanier L , Nagler A , Greenberg P 
Published: October 2019

Abstract
Because T lymphocytes and natural killer (NK) cells produce a variety of growth factors and interleukin 2 (IL2) modulates the activity of both, we assessed the ability of IL2 to stimulate human T cells and NK cells to produce hematopoietic growth factors detectable in clonogenic marrow culture. Human recombinant interleukin 2 (rIL2) added directly to cultures of human bone marrow that had been depleted of monocytes or depleted of both monocytes and T cells caused no significant alteration of myeloid (CFU-GM) or erythroid colony formation. Conditioned media harvested from rIL2-stimulated (greater than 100 U/mL) peripheral blood mononuclear cells, T cells, Leu-2 cells, and Leu-3 cells all had erythroid burst-promoting activity (BPA) but lacked myeloid colony- stimulating factor (GM-CSF) or CFU-GM-inhibitory activity. These T cells were IL2 receptor-negative, and the addition of anti-IL2 receptor monoclonal antibody (anti-Tac) to T cell cultures did not abrogate this IL2-stimulated BPA production. In addition, Percoll gradient-enriched, large granular lymphocytes (LGL) were separated by fluorescence- activated cell sorting into Leu-11+ (NK) cells and Leu-11- (low-density Leu-4+ T) cell fractions. rIL2 stimulated LGL, Leu-11+ and Leu-11- cells to produce BPA but not detectable GM-CSF or CFU-GM-inhibitory activity. Leu-11+ (NK) cells were Tac-negative from days 0 through 14 of culture. We conclude that rIL2 at high concentrations stimulated T cells, Leu-2 and Leu-3 cell subsets, LGL, and NK cells to produce BPA but not GM-CSF and that this stimulation may be mediated by an IL2 receptor distinct from Tac or by an epitope of the IL2 receptor not recognized by the anti-Tac antibody.
Slow disappearance of peripheral blast cells: an independent risk factor indicating poor prognosis in children with acute lymphoblastic leukemia
Authors: Rautonen J , Hovi L , Siimes MA 
Published: October 2019

Abstract
The aim of this study was to find out whether the time required for disappearance of peripheral blast cells, or blast clearance, could be used to identify patients with a slow response to treatment associated with a poor prognosis of acute lymphoblastic leukemia (ALL). Our series consisted of 158 children with newly diagnosed ALL. The mean follow-up time was 69 months (range 22 to 140 months). Blast clearance was significantly associated with length of event-free survival. Only two of nine children with blast clearance greater than or equal to 2 weeks and 4 of 11 children with blast clearance of 11 to 13 days were in remission at the time of analysis as compared with 86 of 138 of the children with more rapid blast clearance. The respective 5-year event- free survivals were 17%, 36%, and 60% (P = .003). Multivariate analysis showed that the relative risk of death or relapse in patients with blast clearance of greater than 10 days was 5.2-fold (95% confidence limits 2.1 to 13.1) as compared with the others (P less than .001). Our results indicate that patients with a slow response to treatment can be identified by simple differential peripheral cell counts during the early induction phase well before or even instead of performance of a more invasive bone marrow aspiration.
Granulocyte/macrophage colony-stimulating factor stimulates monocyte and tissue macrophage proliferation and enhances their responsiveness to macrophage colony-stimulating factor
Authors: Chen BD , Clark CR , Chou TH 
Published: October 2019

Abstract
Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a specific humoral growth factor that stimulates both neutrophilic granulocyte and macrophage production by bone marrow hematopoietic progenitor cells. GM-CSF also stimulates the proliferation and clonal growth of both tissue macrophages and blood monocytes. Although at low concentrations GM-CSF was unable to support the long-term growth of tissue macrophages, it greatly enhanced their responsiveness to macrophage CSF (M-CSF, or CSF-1). This effect was also observed by treating macrophages with GM-CSF for a short time. GM-CSF did not compete with M-CSF for binding to M-CSF receptors nor was it inactivated by treatment with anti-M-CSF antiserum. Treatment of tissue macrophages with GM-CSF led to a rapid but transient downregulation of M-CSF receptors; prolonged incubation at 37 degrees C, however, resulted in a restoration and upregulation of M-CSF receptors. Identical effects were observed with both native or recombinant GM-CSF. This study suggests that GM-CSF regulates tissue macrophage production by two modes of action: (a) direct stimulation of macrophage proliferation, and (b) enhancement of their responsiveness to M-CSF.
Flow cytometric analysis of normal human megakaryocytes
Authors: Tomer A , Harker LA , Burstein SA 
Published: October 2019

Abstract
Megakaryocytes from normal routine human bone marrow aspirates were analyzed by flow cytometry for size, fine cell structure and granularity, membrane expression of glycoprotein (GP) IIb/IIIa and ploidy. Marrow cells were initially enriched for megakaryocytes by a Percoll density gradient and megakaryocytes were labeled with a fluoresceinated monoclonal antibody directed to the GPIIb/IIIa complex. The cells were fixed with paraformaldehyde and stained with propidium iodide (PI) for DNA quantitation. Using two-color flow cytometry, megakaryocytes were identified by their high membrane immunofluorescence and their ploidy was determined according to the relative fluorescence intensity of the PI. Forward light scatter (FSC), correlating with cell size, 90 degrees side light scatter (SSC), reflecting primarily cell internal fine structure and granularity, and total cell membrane fluorescence were examined. To evaluate independently the relationship between size and cell membrane fluorescence obtained by flow cytometry, megakaryocytes were sorted directly on slides and analyzed by a laser-based anchored cell analyzer (ACAS). There was a strong correlation among size, SSC, and the level of membrane fluorescence. The mean diameter of megakaryocytes was 28.1 +/- 12.3 micron. The modal ploidy distribution was 16N with approximately one-fifth of the cells less than or equal to 4N. The mean FSC and SSC levels increased with increasing ploidy. However, the marked overlap observed between the ranges of these parameters in adjacent ploidy classes suggested that size and SSC increase continuously rather than by discrete steps as is characteristic for ploidy. The total surface membrane fluorescence was correlated with cell size (r = 0.98) as measured by FSC or directly by the ACAS (r = 0.85), and with cell ploidy (r = 0.99) indicating an augmentation in total membrane GPIIb/IIIa expression with an increase in cell size and ploidy. However, estimated GPIIb/IIIa fluorescence density was inversely correlated with FSC suggesting that the GPIIb/IIIa surface epitope density is decreased with increasing cell maturity. We conclude that flow cytometry is a useful technique for the rapid analysis of human megakaryocytes obtained by marrow aspiration, and should be applicable to studies of pathologic states.