bluuebunny/crossref_metadata_2025_split huggingface dataset:
Posttranslational processing of a human myeloid lysosomal protein, myeloperoxidase
Authors: Nauseef WM 
Published: October 2019

Abstract
Myeloperoxidase (MPO) is a lysosomal enzyme present in the azurophilic granules of human neutrophils and monocytes and is important for optimal oxygen-dependent killing of microorganisms. The native molecule is a heterodimer composed of a pair of heavy-light protomers, each containing a 59-kDa and 13.5-kDa subunit. The intracellular processing during biosynthesis of MPO was examined in the human promyelocytic cell line HL-60. Endoglycosidase H and F digestion of immunoprecipitated pro- MPO demonstrated the presence of five N-linked--high-mannose oligosaccharide side chains and no complex mannose units. Incorporation of the threonine analogue beta-hydroxynorvaline produced species approximately 2.5 kDa and approximately 5 kDa smaller than the fully glycosylated pro-MPO, suggesting that two of the glycans were in the asparagine-X-threonine tripeptide sequence. Processing of pro-MPO occurred very rapidly, within approximately five minutes, and was best identified using glucosidase inhibitors. The presence of such inhibitors resulted in synthesis of a 92-kDa glycoprotein rather than the usually identified 89-kDa peptide. Swainsonine, a Golgi mannosidase inhibitor, did not alter the size of the earliest synthesized protein, suggesting that pro-MPO exited the endoplasmic reticulum or cis-Golgi proximal to the site of mannosidases. Intracellular transport and proteolytic maturation of MPO was retarded by weak bases (NH4Cl, chloroquine) or monensin at concentrations shown to raise intralysosomal pH. However, these agents did not qualitatively alter transport nor increase secretion. Thus, although MPO biosynthesis resembled that of other lysosomal enzymes, significant differences exist, including only limited oligosaccharide processing and intracellular transport and proteolytic maturation of pro-MPO that was only retarded by alkalinizing lysosomes without affecting the products or the fraction of pro-MPO secreted. Characterization of the determinants for targeting and of the regulatory factors in processing lysosomal enzymes in myeloid cells will provide insight into the molecular mechanisms underlying common disorders such as myeloperoxidase deficiency.
Relationship between T200 antigen expression and stages of B cell differentiation in resurgent hyperplasia of bone marrow
Authors: Caldwell CW , Patterson WP 
Published: October 2019

Abstract
Using monoclonal antibodies (MoAbs) and dual-parameter flow cytometric techniques, bone marrow mononuclear cells (MMC) from patients with resurgent hyperplasia were analyzed for their coexpression of HLe-1 (T200) and antigens normally associated with particular stages of B cell differentiation. The marrow from those with resurgent hyperplasia contained increased numbers of B cell precursors in multiple stages of differentiation compared to controls, thus providing a useful model system for studies of B cell differentiation. These studies indicate that the quantitative expression of T200 is differentiation-related on normal and malignant B cells and B cell precursors. Immature cells express low amounts of T200, while increasing levels of maturity correlated with increasing amounts of the antigen. This study increases the understanding of relationships between B cell surface antigens and T200 and further demonstrates that B cell hyperplasia occurs commonly in association with bone marrow reactive or resurgent processes. The quantitative, rather than only the qualitative, expression of T200 is therefore a useful marker of B cell differentiation in reactive hyperplasia and in further investigation of B cell malignancy.
Effect of recombinant interferons alpha and gamma on human bone marrow- derived megakaryocytic progenitor cells
Authors: Ganser A , Carlo-Stella C , Greher J , Volkers B , Hoelzer D 
Published: October 2019

Abstract
Interferons (IFNs) have been shown to suppress the proliferation of human pluripotent hematopoietic progenitor cells, CFU-GEMM, and committed erythroid (BFU-E, CFU-E) and granulocyte-macrophage (CFU-GM) progenitor cells. However, no information is yet available concerning the effect of IFNs on human megakaryocytic progenitor cells CFU-Mk. Furthermore the mechanisms underlying the inhibitory activity of IFNs are still controversial. Therefore highly purified recombinant IFN preparations, rIFN-alpha and rIFN-gamma, were assessed for their influence on in vitro growth of human bone marrow-derived CFU-Mk as well as CFU-GEMM. In addition, the role of hematopoietic accessory cells, that is, adherent cells and T lymphocytes, in the mediation of the suppressive effect of rIFNs was examined. When added to unseparated bone marrow cells, both rIFN preparations significantly inhibited colony formation with 50% inhibition of CFU-Mk occurring at 22 U/mL for rIFN-alpha and 59 U/mL for rIFN-gamma, while 50% inhibition of CFU-GEMM occurred at 59 U/mL for rIFN-alpha and 101 U/mL for rIFN-gamma. The suppressive effect of rIFN-alpha and rIFN-gamma was selectively abolished by monoclonal antibodies (MoAbs) against rIFN-alpha and rIFN- gamma, thus confirming that the inhibitory activity was due to the rIFN preparations used. The antiproliferative effect of rIFN-alpha and rIFN- gamma on CFU-GEMM growth was not associated with a decrease in the percentage of mixed colonies containing megakaryocytic cells as assessed by use of the MoAb C17.28 against platelet glycoprotein IIIa. Removal of adherent cells and T lymphocytes from the target bone marrow cells had no influence on the suppressive effect of rIFN-alpha, whereas it significantly reduced the inhibitory effect of rIFN-gamma on the growth of megakaryocytic colonies and the other hematopoietic progenitors. The data indicate that (1) human megakaryocytopoiesis is markedly inhibited by rIFN-alpha and rIFN-gamma, and (2) the inhibitory effect of rIFN-alpha is due to a direct action on hematopoietic progenitor cells, whereas the effect of rIFN-gamma is mediated to a significant degree through accessory cell populations.
Chloramphenicol-induced bone marrow injury: possible role of bacterial metabolites of chloramphenicol
Authors: Jimenez JJ , Arimura GK , Abou-Khalil WH , Isildar M , Yunis AA 
Published: October 2019

Abstract
To explore the potential role of some bacterial metabolites of chloramphenicol (CAP) in CAP-induced hematotoxicity, we examined their cytotoxic effects on bone marrow cells in vitro using a number of cytotoxicity parameters. Among the metabolites tested, dehydro-CAP (DHCAP) and p-nitrophenyl-2-amino-3 hydroxypropanone-HCI (NPAP) were more toxic than CAP. DHCAP was at least as toxic as nitroso-CAP. At concentrations of less than or equal to 10(-4) mol/L, DHCAP caused total irreversible inhibition of myeloid colony (CFU-GM) growth and 80% inhibition of DNA synthesis in human bone marrow. Incubation of human bone marrow cells with 10(-4) mol/L nitroso-CAP or DHCAP for 24 hours resulted in 75% and 65% cell death respectively. Although DHCAP was 10- to 20-fold more cytotoxic than CAP, it was only one third as effective in inhibiting mitochondrial protein synthesis, indicating that DHCAP exerts its toxic effect by alternate mechanisms. The cytotoxicity of DHCAP and its relative stability, compared to the unstable nitroso CAP, suggest that this bacterial metabolite of CAP, and possibly others, may play a significant role in CAP-induced hematotoxicity.
Human leukemic B cell activation: functional consequence of membrane IgM interaction with anti-IgM ligand is an alterable cell characteristic
Authors: Mongini P , Blessinger C , Seremetis S , Winchester R , Rudich S 
Published: October 2019

Abstract
A functional study of several human malignant B cell populations has indicated that occasional leukemic clones are extraordinarily sensitive to signal transduction through membrane IgM. One isolated hairy cell leukemia (HCL) with low background DNA synthesis was stimulated to significant levels of DNA synthesis when cultured with high (100 micrograms/mL) concentrations of soluble anti-IgM ligands. In contrast to the activation of normal peripheral blood polyclonal B cells, this DNA synthesis was completely independent of accessory T cell factors. Although the HCL clone could also be induced to enter S phase by incubation in media supplemented with only activated T cell supernatant, culture of the clone with activated T cell supernatant plus anti-IgM Ab resulted in DNA synthesis that was significantly less than that induced by either activator alone. Factor(s) in T cell supernatant appear to modulate the leukemic clone so that the binding of ligand to membrane IgM is perceived as an inhibitory rather than a stimulatory signal for DNA synthesis. In terms of Ig Fc independence and low ligand dose requirements, anti-IgM-mediated inhibitory signal transduction in the T cell supernatant-activated HCL clone was found to mimic anti-IgM mediated suppression of the spontaneous DNA synthesis of an alternative HCL clone. The functional results suggest that the type of signal transduced anti-Ig ligands may reflect differences in the activation state of receptive leukemic B cells.
Therapy of chronic myelogenous leukemia with recombinant interferon- gamma
Authors: Kurzrock R , Talpaz M , Kantarjian H , Walters R , Saks S , Trujillo JM , Gutterman JU 
Published: October 2019

Abstract
Recently, we reported that recombinant interferon-alpha (rIFN-alpha) can induce hematologic remissions and cytogenetic improvement in newly diagnosed Philadelphia (Ph)-positive chronic myelogenous leukemia (CML) patients. Although IFN-gamma is a structurally distinct molecule, this agent suppresses in vitro hematopoietic progenitor cells in a fashion similar to that of IFN-alpha. Therefore, we initiated a study of rIFN- gamma at doses of 0.25 to 0.5 mg/m2/d intramuscularly in patients with Ph-positive benign-phase CML. Twenty-six of 30 patients entered in the study were evaluable. Six patients have achieved a complete hematologic response; four, a partial hematologic response. The median follow-up period of patients who are in complete remission is 7.5 months (range, 5 to 12 months). No relapses have occurred among the complete responders. So far, five patients have had cytogenetic improvement with emergence of 5% to 45% diploid cells in the bone marrow. Fever and flulike symptoms were the most common side effects, with partial tolerance often developing after about 1 week. The majority of patients tolerated therapy with minimal change in performance status. In conclusion, rIFN-gamma has demonstrated clinical activity in CML. On the basis of these observations and the in vitro synergistic growth- inhibitory effects of IFN-alpha and IFN-gamma, we have started trials of combination IFN therapy in CML patients.
Autologous bone marrow transplantation using unfractionated cells cryopreserved in dimethylsulfoxide and hydroxyethyl starch without controlled-rate freezing
Authors: Stiff PJ , Koester AR , Weidner MK , Dvorak K , Fisher RI 
Published: October 2019

Abstract
To develop a simplified method of bone marrow (BM) cryopreservation, changes were made in the standard method in three areas: the cryoprotectant, the method of cell freezing, and the storage temperature. Unfractionated BM cells from 60 patients were cryopreserved in 300-mL aliquots in both dimethylsulfoxide (DMSO) and hydroxyethyl starch (HES), a combination known to preserve granulocytes successfully. The cells were frozen without rate-controlled freezing by simple immersion into a -80 degrees C freezer where they remained until the time of reinfusion. The 60 patients underwent 72 autologous transplants after three high-dose chemotherapy regimens: 30 received high-dose carmustine in combination, five received high-dose busulfan and cyclophosphamide, and 37 received high-dose aziridinylbenzoquinone. The BM was infused for more than 30 minutes after rapid thawing at 37 degrees C. The mean post-thaw nucleated cell recovery was 96% +/- 11.6%, and Trypan blue dye exclusion was 82.2% +/- 9.2%. The mean postthaw CFU-GM and BFU-E recoveries were 81.9% +/- 39.0% and 90.5% +/- 41.2%. Complete count recovery occurred in 68 of 72 transplants. Median times to a WBC count greater than 1,000/microL, a granulocyte count greater than 1,000/microL and a platelet count greater than 20,000/microL were 15, 21, and 15 days, respectively. Risk factors for delayed recovery were not found. Unfractionated BM cells can be successfully cryopreserved in the DMSO/HES mixture rapidly and inexpensively, without rate-controlled freezing or storage at liquid nitrogen temperatures.
Isolation of lactoferrin cDNA from a human myeloid library and expression of mRNA during normal and leukemic myelopoiesis
Authors: Rado TA , Wei XP , Benz EJ Jr 
Published: October 2019

Abstract
Lactoferrin is a major constituent of polymorphonuclear leukocyte granules and is present in mature neutrophils but not in blasts or promyelocytes. We have isolated a cDNA probe for lactoferrin and used it to study the synthesis of lactoferrin mRNA by normal and leukemic granulocyte precursors. The probe pHL-41 has been subcloned in phage m13 and characterized by restriction endonuclease analysis and nucleic acid sequencing. pHL-41 contains approximately 40% of the coding sequence of the lactoferrin gene. The 3′ untranslated region includes a stop codon and a possible polyadenylation signal. There is a greater than 98% agreement between the cDNA-deduced amino acid sequence and that determined by analysis of the protein. Myeloid cells from normal bone marrow and circulating leukocytes from patients with chronic granulocytic leukemia contain lactoferrin mRNA transcripts that are indistinguishable in size and relative quantity. The human promyelocytic leukemia cell line HL-60 contains no lactoferrin mRNA. Induction of monocytic or granulocytic differentiation fails to induce the synthesis of detectable lactoferrin message. Similarly, studies with the human myeloblastic leukemia cell line PLB-985 reveal the inability of these cells to produce lactoferrin mRNA even under conditions that bring about morphologically demonstrable granulocytic differentiation. These data suggest that granulocytic differentiation in the leukemic cell lines is incomplete or defective. The presence of lactoferrin may play a role in the orderly expression of the genetic program leading to the development of the normal mature granulocyte.
The HL-60 promyelocytic leukemia cell line: proliferation, differentiation, and cellular oncogene expression
Authors: Collins SJ 
Published: October 2019

Abstract
The HL-60 cell line, derived from a single patient with acute promyelocytic leukemia, provides a unique in vitro model system for studying the cellular and molecular events involved in the proliferation and differentiation of normal and leukemic cells of the granulocyte/monocyte/macrophage lineage.
Molecular genetic survey of 16 kindreds with hereditary antithrombin III deficiency
Authors: Bock SC , Prochownik EV 
Published: October 2019

Abstract
Molecular genetic techniques were utilized to examine antithrombin III (ATIII) gene status in 16 independently ascertained kindreds with hereditary ATIII deficiency. In one of these families antithrombin III deficiency is caused by hemizygosity of the ATIII locus. In the remaining 15 kindreds, two copies of the ATIII gene are present and appear to be grossly normal at the level of whole genome Southern blotting, suggesting that small deletions, insertions or limited nucleotide substitution(s) in the antithrombin III gene, or “trans- acting” defects at other loci involved in the processing, modification, and secretion of biologically active ATIII are responsible for the observed anticoagulant disorders.
Effect of flow on polymorphonuclear leukocyte/endothelial cell adhesion
Authors: Lawrence MB , McIntire LV , Eskin SG 
Published: October 2019

Abstract
The effect of flow on the adhesion of polymorphonuclear leukocytes (PMNL) to vascular endothelium was investigated using a parallel plate chamber with a well-defined flow field. Washed PMNL were perfused over a monolayer of primary human umbilical vein endothelial cells (HUVEC) pretreated with formyl-methionyl-leucyl-phenylalanine (FMLP, 1 X 10(-7) mol/L) for five minutes. In other experiments HUVEC were pretreated with interleukin 1 (IL1,2 U/mL) for four hours. PMNL adhesion to stimulated and control HUVEC was measured over a physiologic range of wall shear stresses. PMNL adhesion to nylon-coated surface was also studied. At a wall shear stress of 0.98 dynes/cm2,283 +/- 37.3 PMNL/mm2 (mean +/- SEM) adhered to FMLP-treated HUVEC while 195 +/- 20.3 PMNL/mm2 adhered to control HUVEC. At 1.96 dynes/cm2, 68 +/- 14.1 PMNL/mm2 adhered to FMLP-treated HUVEC and 42 +/- 6.0 PMNL/mm2 adhered to control HUVEC. At 3.92 dynes/cm2, virtually no PMNL adherence was noted on either control or FMLP-treated HUVEC. On IL 1-treated HUVEC at 1.96 dynes/cm2, 371 +/- 25.8 PMNL/mm2 adhered while 28 +/- 2.9 PMNL/mm2 adhered to control HUVEC. PMNL adhesion to IL 1-treated and control HUVEC dropped to 10.2 +/- 3.8 and 6.8 +/- 3.5 PMNL/mm2, respectively, at 3.01 dynes/cm2. The effect of flow on PMNL adhesion appears to be an important factor in determining the outcome of the PMNL/HUVEC adhesive interaction under these experimental conditions.
Cytogenetic and cytochemical studies on progenitor cells of primary acquired sideroblastic anemia (PASA): involvement of multipotent myeloid stem cells in PASA clone and mosaicism with normal clone
Authors: Amenomori T , Tomonaga M , Jinnai I , Soda H , Nonaka H , Matsuo T , Yoshida Y , Kuriyama K , Ichimaru M , Suematsu T 
Published: October 2019

Abstract
By cytogenetic and cytochemical analyses of individual hematopoietic colonies, we investigated clonality in progenitor compartments of primary acquired sideroblastic anemia (PASA). Two of our four subjects had reduced but countable numbers of CFU-E, BFU-E, and GFU-GM in methylcellulose culture. In one patient with cytogenetic abnormality of 47, XX, +8 in 67% of the bone marrow cells, cytogenetic analysis of individual erythroid bursts and granulocyte/macrophage colonies demonstrated two populations with and without 8 trisomy, the trisomy clone being 38% in BFU-E and 50% in CFU-GM. These findings indicate involvement of multipotent stem cells in PASA clone and mosaicism of two distinct populations in erythroid as well as granulocyte/macrophage progenitor compartments, the abnormal PASA clone and probably the normal clones. In another case with no cytogenetic abnormality, repeated iron staining showed that 31% to 40% of CFU-E and 25% to 54% of BFU-E had erythroblasts with heavy iron deposits. An ultrastructural analysis of 25 individual erythroid bursts revealed that 32% had highly dysplastic erythroblasts with marked ferruginous iron accumulation in the mitochondria. The other 68% and 15 normal bursts from a healthy control did not have noticeable dysplastic changes and iron deposits in the mitochondria. This cytochemical/ultrastructural mosaicism seems to be compatible with the cytogenetic mosaicism. However, whether the BFU- E derived from abnormal PASA clone selectively manifest iron accumulation in the mitochondria or whether the PASA clone itself shows variable degrees of abnormal iron metabolism remains to be determined by simultaneous performance of ultrastructural and cytogenetic analysis for single bursts.
Minimally differentiated acute nonlymphocytic leukemia: a distinct entity
Authors: Lee EJ , Pollak A , Leavitt RD , Testa JR , Schiffer CA 
Published: October 2019

Abstract
Ten of 136 consecutive adult patients with previously untreated acute leukemia had morphologically undifferentiated leukemia by light microscopy. Leukemic cells from these patients were characterized by agranular cytoplasm, negative histochemical staining with sudan black (SB) and nonspecific esterase, and absent lymphoid cell surface markers and therefore were not classifiable according to the French-American- British (FAB) system. Electron microscopy with myeloperoxidase (MPO) staining revealed the presence of peroxidase positive cytoplasmic granules and endoplasmic reticulum in eight of the nine patients studied. Cells from the patient who was negative for MPO were also negative for platelet peroxidase. A series of monoclonal antibodies to myeloid antigens also revealed myeloid features with all patients having at least one myeloid differentiation antigen present on the surface of their cells. Common acute lymphoblastic leukemia (ALL) antigen was absent in the nine patients tested. Cytogenetic analysis of blast cells was abnormal in seven patients on whom adequately banded chromosomes were obtained although there were no consistent abnormalities. No patient had a Ph1 chromosome. Only two of the ten patients achieved a complete remission. Morphologically undifferentiated leukemia may have myeloid features when studied by transmission electron microscopy or with monoclonal antibodies for cell surface markers. Such studies should be performed when the leukemia cannot be classified using either light microscopy or lymphoid cell surface markers. Such patients infrequently achieve remission with standard therapy and constitute a distinct entity.
B lymphocyte precursors and myeloid progenitors survive in diffusion chamber cultures but B cell differentiation requires close association with stromal cells
Authors: Kierney PC , Dorshkind K 
Published: October 2019

Abstract
The aim of this study was to investigate the relative contribution of direct contact with stromal cells v stromal cell-derived soluble mediators to the differentiation of B lymphocytes and cells from other hematopoietic lineages. This was investigated by making a comparison between hemopoietic cells grown in direct contact with stroma to those in diffusion chambers (DCs) placed over purified populations of stroma. The source of stromal cells was adherent layers from myeloid or lymphoid long-term bone marrow cultures that had been treated with mycophenolic acid, an antibiotic that depletes hemopoietic cells from the cultures but retains a functional stroma. The cells seeded into the chambers were fresh marrow cells that had been passed through two consecutive nylon wool columns to deplete cell populations capable of forming an adherent cell layer in vitro. DCs were placed in wells in which the adherent stroma, growing under myeloid or lymphoid conditions, was present. The results indicate that progenitors of granulocytes and macrophages survived and differentiated in DCs under myeloid culture conditions, as the number of cells and absolute number of CFU-GM increased over that present in the reseed population. These levels, however, were markedly less than in parallel cultures in which the cells were seeded directly onto stroma. Hematopoiesis in DCs placed over hemopoietically active stroma was not optimal, suggesting that factors were used by those hemopoietic cells closest to the stroma. A B lymphocyte precursor survived in DCs under myeloid but not lymphoid conditions, and its differentiation into B lymphocytes was dependent on close association with stromal cells; B lymphopoiesis initiated when cells from DCs grown under myeloid conditions were harvested from the chambers and seeded directly onto stroma initiated and maintained under lymphoid bone marrow culture conditions. B lymphopoiesis did not initiate if the DC from the myeloid conditions was left intact and placed directly over a lymphoid stromal cell layer in lymphoid conditions.