bluuebunny/crossref_metadata_2025 huggingface dataset:
Tissue factor pathway inhibitor dose-dependently inhibits coagulation activation without influencing the fibrinolytic and cytokine response during human endotoxemia
Authors: de Jonge Evert , Dekkers Pascale E. P. , Creasey Abla A. , Hack C. Erik , Paulson Susan K. , Karim Aziz , Kesecioglu Jozef , Levi Marcel , van Deventer Sander J. H. , van der Poll Tom
Published: October 2019

Inhibition of the tissue factor pathway has been shown to attenuate the activation of coagulation and to prevent death in a gram-negative bacteremia primate model of sepsis. It has been suggested that tissue factor influences inflammatory cascades other than the coagulation system. The authors sought to determine the effects of 2 different doses of recombinant tissue factor pathway inhibitor (TFPI) on endotoxin-induced coagulant, fibrinolytic, and cytokine responses in healthy humans. Two groups, each consisting of 8 healthy men, were studied in a double-blind, randomized, placebo-controlled crossover study. Subjects were studied on 2 different occasions. They received a bolus intravenous injection of 4 ng/kg endotoxin, which was followed by a 6-hour continuous infusion of TFPI or placebo. Eight subjects received 0.05 mg/kg per hour TFPI after a bolus of 0.0125 mg/kg (low-dose group), and 8 subjects received 0.2 mg/kg per hour after a bolus of 0.05 mg/kg (high-dose group). Endotoxin injection induced the activation of coagulation, the activation and subsequent inhibition of fibrinolysis, and the release of proinflammatory and antiinflammatory cytokines. TFPI infusion induced a dose-dependent attenuation of thrombin generation, as measured by plasma F1 + 2 and thrombin–antithrombin complexes, with a complete blockade of coagulation activation after high-dose TFPI. Endotoxin-induced changes in the fibrinolytic system and cytokine levels were not altered by either low-dose or high-dose TFPI. The authors concluded that TFPI effectively and dose-dependently attenuates the endotoxin-induced coagulation activation in humans without influencing the fibrinolytic and cytokine response.
KSHV-encoded CC chemokine vMIP-III is a CCR4 agonist, stimulates angiogenesis, and selectively chemoattracts TH2 cells
Authors: Stine Johnny T. , Wood Christi , Hill Mark , Epp Angela , Raport Carol J. , Schweickart Vicki L. , Endo Yoshio , Sasaki Takuma , Simmons Graham , Boshoff Chris , Clapham Paul , Chang Yuan , Moore Patrick , Gray Patrick W. , Chantry David
Published: October 2019

Kaposi's sarcoma-associated herpesvirus (KSHV) encodes 3 genes that are homologous to cellular chemokines. vMIP-III, the product of open reading frame K4.1, is the most distantly related to human chemokines and has yet to be characterized. We have examined the interaction of vMIP-III with chemokine receptors, its expression in KS lesions, and its in ovo angiogenic properties. We show expression of vMIP-III in KS lesions and demonstrate the stimulation of angiogenesis by this chemokine, like vMIP-I and vMIP-II, in the chick chorioallantoic membrane assay. vMIP-III does not block human immunodeficiency virus entry through the coreceptors CCR3, CCR5, or CXCR4. However, vMIP-III is an agonist for the cellular chemokine receptor CCR4. CCR4 is expressed by TH2-type T cells. Consistent with this, vMIP-III preferentially chemoattracts this cell type. Because of these biologic properties and because it is expressed in KS lesions, vMIP-III may play an important role in the pathobiology of KS.
Allogeneic and syngeneic marrow transplantation for myelodysplastic syndrome in patients 55 to 66 years of age
Authors: Deeg H. J. , Shulman H. M. , Anderson J. E. , Bryant E. M. , Gooley T. A. , Slattery J. T. , Anasetti C. , Fefer A. , Storb R. , Appelbaum F. R.
Published: October 2019

We carried out bone marrow transplantation (BMT) in 50 patients with myelodysplastic syndrome (MDS) who were 55.3 to 66.2 years of age (median, 58.8 years). According to the criteria of the French-American-British (FAB) classification, 13 patients had refractory anemia (RA), 19 had RA with excess blasts (RAEB), 16 had RAEB in transformation or acute myelogenous leukemia (RAEB-T/AML), and 2 had chronic myelomonocytic leukemia (CMML). According to the recently established International Prognostic Scoring System (IPSS), available for 45 patients, 2 patients were considered low risk; 14, intermediate 1 risk; 19, intermediate 2 risk; and 10, high risk. Conditioning regimens were cyclophosphamide (CY) (120 mg/kg of body weight) plus 12-Gy fractionated total-body irradiation (FTBI) (n = 15), CY plus FTBI with lung and liver shielding (n = 4), busulfan (7 mg/kg) plus FTBI (n = 4), or busulfan (16 mg/kg) plus CY (n = 27). The busulfan-plus-CY group included 16 patients in whom busulfan was targeted to plasma levels of 600 to 900 ng/mL. In these 16 patients, steady-state levels of busulfan actually achieved were 714 to 961 ng/mL (mean ± SD, 845 ± 64 ng/mL; median, 838 ng/mL). The donors were HLA-identical siblings for 34 patients, HLA-nonidentical family members for 4, identical twins for 4, and unrelated volunteers for 6. All 46 patients surviving > 21 days had engraftment, and 22 patients (44%) are surviving 9 to 80 months after BMT. Specifically, among 13 patients with RA, 1 had relapse (cumulative incidence [CI] at 3 years, 8%) and 8 are surviving, for a Kaplan-Meier (KM) estimate of survival at 3 years of 59% (disease-free survival [DSF], 53%). Among 19 patients with RAEB, 3 had relapse (CI at 3 years, 16%), and 8 are surviving disease free (KM estimate at 3 years, 46%). Among 18 patients with RAEB-T/AML or CMML, 6 had relapse (CI at 3 years, 28%), and the KM estimate of DSF at 3 years is 33%. Relapse-free survival had an inverse correlation with cytogenetic risk classification and with the risk score according to the IPSS. Survival in all FAB categories was highest among patients enrolled in a protocol in which busulfan plasma levels were targeted to 600 to 900 ng/mL. These data indicate that BMT can be carried out successfully in patients with MDS who are older than 55 years of age.
Apoptosis or plasma cell differentiation of CD38-positive B-chronic lymphocytic leukemia cells induced by cross-linking of surface IgM or IgD
Authors: Zupo Simona , Massara Rosanna , Dono Mariella , Rossi Edoardo , Malavasi Fabio , Cosulich M. Elisabetta , Ferrarini Manlio
Published: October 2019

Previously, we demonstrated that B-chronic lymphocytic leukemia (B-CLL) cells could be divided into 2 groups depending on the expression of CD38 by the malignant cells. The 2 groups differed in their signal-transducing capacities initiated by cross-linking of surface IgM; only in CD38-positive cells was an efficient signal delivered, invariably resulting in cell apoptosis. In this study, we investigated the effect of surface IgD cross-linking in 10 patients with CD38-positive B-CLL. Exposure of the malignant cells to goat antihuman δ-chain antibodies (Gaδ-ab) caused [Ca++]i mobilization and tyrosine kinase phosphorylation in a manner not different from that observed after goat antihuman μ-chain antibody (Gaμ-ab) treatment in vitro. However, Gaδ-ab-treated cells failed to undergo apoptosis and instead displayed prolonged survival in culture and differentiated into plasma cells when rIL2 was concomitantly present. Cross-linking of surface IgD failed to induce proliferation of the malignant cells in vitro. Moreover, treatment with Gaδ-ab did not prevent apoptosis of B-CLL cells induced by Gaμ-ab. Collectively, these experiments demonstrated that IgM and IgD expressed by the same cell may deliver opposite signals under particular circumstances and provide some clues for the understanding of the pathophysiology of B-CLL.
Tissue eosinophilia correlates strongly with poor prognosis in nodular sclerosing Hodgkin's disease, allowing for known prognostic factors
Authors: von Wasielewski R. , Seth S. , Franklin J. , Fischer R. , Hübner K. , Hansmann M. L. , Diehl V. , Georgii A.
Published: October 2019

Although eosinophilic granulocytes are frequently observed in lymphatic tissue of Hodgkin's patients, no substantial data reveal the prognostic role, if any, of tissue eosinophilia. Thus, eosinophilia was analyzed histologically in 1511 diagnostic biopsy specimens of patients treated under protocol therapy of the German Hodgkin's Lymphoma Study Group between 1988 and 1994. Prominent eosinophilia was seen in 38% of cases, which differed among the histologic types of Hodgkin's disease (HD): none in lymphocyte predominant, 14% in lymphocyte rich classical, 40% in nodular sclerosis grade 1 (NS-1), 55% in nodular sclerosis grade 2, 43% in mixed cellularity (MC), and 54% in lymphocyte depleted. In a multivariate analysis, tissue eosinophilia proved to be the strongest prognostic factor for freedom from treatment failure (P < .001) and overall survival (P < .001) in a stage-stratified model. Among NS-1 patients, the effect was highly significant. In MC, no significant effect of eosinophilia on survival could be demonstrated. Eosinophils secrete CD30 ligand that is capable of binding to CD30 positive HD cells. The activation of TRAF2, followed by NF-kappaB, which occurs on CD30L/CD30 binding, may explain the neoplastic proliferation and apoptosis protection of HD cells. TRAF2 is also activated by EBV-LMP expression, which is detectable in the majority of MC but not NS cases. In addition to the possibility that eosinophils are only passive indicators for other unknown prognostic determinants, it may be concluded that the positive clinical outcome of eosinophilia-negative NS cases could be due to lower NF-kappaB activity.
Targeted disruption of Stat6 DNA binding activity by an oligonucleotide decoy blocks IL-4–driven TH2 cell response
Authors: Wang Li Hua , Yang Xiao Yi , Kirken Robert A. , Resau James H. , Farrar William L.
Published: October 2019

The transcription factor, signal transducer and activator of transcription (Stat) 6, regulates TH2-lymphocyte activity by controlling the expression and responsiveness to interleukin (IL)–4, which plays a key role in numerous allergic maladies. Therefore, we sought to use a phosphorothiolate cis-element decoy to target disruption of Stat6 transcriptional activity. Here we showed that the Stat6 decoy potently ablated the messenger RNA expression and production of IL-4, but not of several other cytokines. The Stat6 decoy functionally disrupted IL-4–inducible cell proliferation of murine TH2 cells and primary human CD4+ T lymphocytes. Specificity of the decoy was demonstrated by its ability to directly block Stat6 binding to a cis-element probe and transactivation, but not affect Stat6 tyrosine phosphorylation or expression of the IL-4 receptor chains. Moreover, the decoy failed to inhibit non–Stat6-dependent signaling pathways since IL-2 was competent to induce cell proliferation and activation of Stats 1, 3, and 5a/b. With the use of laser scanning confocal microscopy, fluorescently tagged Stat6 decoy was detectable in the cytoplasm and nucleus; however, greater levels of oligonucleotide were present in the latter following IL-4 treatment. Taken together, these data suggest that IL-4–driven TH2 cell activity can be preferentially restricted via targeted disruption of Stat6 by a novel and specific decoy strategy that may possess gene therapeutic potential.
Hemopoietic lineage commitment decisions: in vivo evidence from a transgenic mouse model harboring μLCR-βpro-LacZ as a transgene
Authors: Papayannopoulou Thalia , Priestley Gregory V. , Rohde Alex , Peterson Kenneth R. , Nakamoto Betty
Published: October 2019

A substantial body of published data suggests activation of lineage-specific genes in multipotential hemopoietic cells before their unilineage commitment. Because the behavior and plasticity of cells isolated in vitro away from microenvironmental constraints exercised in vivo may be altered, one wonders whether similar findings can be observed in a physiologic setting in vivo. We used a transgenic mouse model harboring human micro LCR together with β promoter sequences as a transgene to examine activation of lineage-specific programs in vivo. By using LacZ as a reporter, we had the ability to detect, quantitate, and select live cells with different levels of LacZ activation. We found strong expression of LacZ by X-gal staining in 2 lineages—erythroid and megakaryocytic. Activation in the latter was a novel finding not previously observed when similar transgenes were used. We also found activation of μLCR-βpro at low levels in progenitor cells of granulocytic-macrophagic, erythroid, or megakaryocytic lineage detected by in vitro assays, suggesting activation before commitment to a specific lineage pathway. In particular, the expression of LacZ was graded among progenitors, so that in a proportion of them activation occurred only after commitment to erythroid or megakaryocytic lineage. In addition, we found quantitative reduction in LacZ expression between fetal liver and bone marrow-derived cells, the basis of which is unclear. Collectively our data provide in vivo evidence supporting the view that lineage-specific genes are expressed in a graded fashion in pluripotential cells before their irreversible unilineage commitment.
Leukocyte-leukocyte interactions mediated by platelet microparticles under flow
Authors: Forlow Stephen B. , McEver Rodger P. , Nollert Matthias U.
Published: October 2019

Platelet microparticles (PMPs) are released from activated platelets and express functional adhesion receptors, including P-selectin, on their surface. PMP concentrations are elevated in many disorders, and their role in accelerating coagulation has been studied. However, their role in leukocyte aggregation has not been defined. We hypothesized that P-selectin–expressing PMPs bridge leukocytes that express P-selectin glycoprotein ligand-1 (PSGL-1), thereby allowing them to interact under flow conditions. PMPs were isolated from platelet-rich plasma or were generated by activating washed platelets with calcium ionophore. PMPs increased transient adhesion of flowing HL-60 cells or neutrophils to HL-60 cells or neutrophils prebound to the surface of a parallel plate flow chamber. Homotypic neutrophil interactions are initiated by the binding of L-selectin to PSGL-1. However, even when L-selectin function was blocked, PMPs allowed flowing neutrophils to aggregate and to interact with PSGL-1–expressing cells prebound to the surface of the flow chamber. The microparticle-mediated cell interactions occurred at lower shear stresses than those mediated by L-selectin. PMPs may enhance leukocyte aggregation and leukocyte accumulation on selectin-expressing substrates, especially in diseases where the concentration of the particles is elevated.
Circulating forms of intercellular adhesion molecule (ICAM)-1 in mice lacking membranous ICAM-1
Authors: van den Engel Natasja K. , Heidenthal Edmund , Vinke Antje , Kolb Hubert , Martin Stephan
Published: October 2019

Mice deficient in intercellular adhesion molecule-1 (ICAM-1), lacking membranous ICAM-1, show a normal development but abnormalities of inflammatory and immune functions. Although the membrane-bound form of ICAM-1 is not detectable in the mutant strain, circulating ICAM-1 (cICAM) is present in serum from ICAM-1-deficient mice in similar amounts as in serum from wild-type mice. These findings were confirmed in vitro by flow cytometric analysis of lipopolysaccharide-stimulated spleen cells, and cICAM-enzyme-linked immunosorbent assay analysis of supernatants of cultured spleen cells. To analyze for the source of cICAM-1, spleen cell RNA was isolated and ICAM-1 RNA was amplified by reverse transcriptase-polymerase chain reaction using primers binding in the 5′ and 3′ untranslated regions. Different fragments were cloned and sequenced. In wild-type RNA the common 5 domain form of ICAM-1 was identified. In RNA from ICAM-1 mutant mice only 3 smaller fragments were found. Sequencing these fragments identified 3 alternatively spliced isoforms of ICAM-1, lacking 2 or 3 extracellular domains. However, in all spliced fragments the transmembrane domain was included. Therefore, we postulate that circulating forms of ICAM-1 are generated by proteolytic cleavage of membranous ICAM-1. The data indicate that the expression of membranous ICAM-1 and the appearance of circulating forms in serum are independently regulated mechanisms.
HHV-8 is associated with a plasmablastic variant of Castleman disease that is linked to HHV-8–positive plasmablastic lymphoma
Authors: Dupin Nicolas , Diss Tim L. , Kellam Paul , Tulliez Micheline , Du Ming-Qing , Sicard Didier , Weiss Robin A. , Isaacson Peter G. , Boshoff Chris
Published: October 2019

Castleman disease (CD) is a lymphoproliferative disorder of unknown etiology that is associated with the development of secondary tumors, including B-cell lymphoma. Human herpesvirus 8 (HHV-8) (Kaposi's sarcoma–associated herpesvirus) sequences have been described in some cases of multicentric Castleman disease (MCD). Using a monoclonal antibody against an HHV-8–latent nuclear antigen, we show that HHV-8 is specifically associated with a variant of MCD in which HHV-8–positive plasmablasts that show λ light-chain restriction localize in the mantle zone of B-cell follicles and coalesce to form microscopic lymphomas in some cases. Furthermore, we show that the frank plasmablastic lymphoma that develops in patients with this plasmablastic variant of MCD is also positive for HHV-8 and λ light chain. Plasmablastic lymphoma associated with MCD is a new disease entity associated with HHV-8 infection.
Interleukin 5 regulates the isoform expression of its own receptor α-subunit
Authors: Tavernier Jan , Van der Heyden José , Verhee Annick , Brusselle Guy , Van Ostade Xaveer , Vandekerckhove Joël , North Janet , Rankin Sara M. , Kay A. Barry , Robinson Douglas S.
Published: October 2019

The receptor for interleukin 5 (IL-5) consists of a cytokine-specific  chain (IL-5R) and a signaling β chain, which is shared with interleukin 3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF). These 3 cytokines can act in eosinophil development and activation in vitro, but gene deletion or antibody blocking of IL-5 largely ablates eosinophilic responses in models of allergic disease or helminth infection. We investigated factors acting in differential IL-5R gene splicing to generate either the membrane-anchored isoform (TM-IL-5R) which associates with the common β chain to allow IL-5 responsiveness, or a secreted, antagonist variant (SOL-IL-5R). In a murine myeloid cell line (FDC-P1), transfected with minigenes allowing expression of either IL-5R variant, IL-5 itself, but not IL-3 or GM-CSF, stimulated a reversible switch toward expression of TM-IL-5R. A switch from predominantly soluble isoform to TM-IL-5R messenger RNA (mRNA) expression was also seen during IL-5-driven eosinophil development from human umbilical cord blood-derived CD34+ cells; this was accompanied by surface expression of IL-5R and acquisition of functional responses to IL-5. IL-3 and GM-CSF also supported eosinophil development and up-regulation of TM-IL-5R mRNA in this system, but this was preceded by expression of IL-5 mRNA and was inhibited by monoclonal antibody to IL-5. These data suggest IL-5-specific signaling, not shared by IL-3 and GM-CSF, leading to a switch toward up-regulation of functional IL-5R and, furthermore, that IL-3 and GM-CSF-driven eosinophil development is dependent on IL-5, providing an explanation for the selective requirement of IL-5 for expansion of the eosinophil lineage.
Leishmania donovani infection of bone marrow stromal macrophages selectively enhances myelopoiesis, by a mechanism involving GM-CSF and TNF-α
Authors: Cotterell Sara E. J. , Engwerda Christian R. , Kaye Paul M.
Published: October 2019

Alterations in hematopoiesis are common in experimental infectious disease. However, few studies have addressed the mechanisms underlying changes in hematopoietic function or assessed the direct impact of infectious agents on the cells that regulate these processes. In experimental visceral leishmaniasis, caused by infection with the protozoan parasite Leishmania donovani, parasites persist in the spleen and bone marrow, and their expansion in these sites is associated with increases in local hematopoietic activity. The results of this study show that L donovani targets bone marrow stromal macrophages in vivo and can infect and multiply in stromal cell lines of macrophage, but not other lineages in vitro. Infection of stromal macrophages increases their capacity to support myelopoiesis in vitro, an effect mediated mainly through the induction of granulocyte macrophage-colony stimulating factor and tumor necrosis factor-. These data are the first to directly demonstrate that intracellular parasitism of a stromal cell population may modify its capacity to regulate hematopoiesis during infectious disease.
Surface expression and functional characterization of α-granule factor V in human platelets: effects of ionophore A23187, thrombin, collagen, and convulxin
Authors: Alberio L. , Safa O. , Clemetson K. J. , Esmon C. T. , Dale G. L.
Published: October 2019

Factor V (FV) present in platelet -granules has a significant but incompletely understood role in hemostasis. This report demonstrates that a fraction of platelets express very high levels of surface-bound, -granule FV on simultaneous activation with 2 agonists, thrombin and convulxin, an activator of the collagen receptor glycoprotein VI. This subpopulation of activated platelets represents 30.7% ± 4.7% of the total population and is referred to as convulxin and thrombin–induced-FV (COAT-FV) platelets. COAT-FV platelets are also observed on activation with thrombin plus collagen types I, V, or VI, but not with type III. No single agonist examined was able to produce COAT-FV platelets, although ionophore A23187 in conjunction with either thrombin or convulxin did generate this population. COAT-FV platelets bound annexin-V, indicating exposure of aminophospholipids and were enriched in young platelets as identified by the binding of thiazole orange. The functional significance of COAT-FV platelets was investigated by demonstrating that factor Xa preferentially bound to COAT-FV platelets, that COAT-FV platelets had more FV activity than either thrombin or A23187–activated platelets, and that COAT-FV platelets were capable of generating more prothrombinase activity than any other physiologic agonist examined. Microparticle production by dual stimulation with thrombin and convulxin was less than that observed with A23187, indicating that microparticles were not responsible for all the activities observed. These data demonstrate a new procoagulant component produced from dual stimulation of platelets with thrombin and collagen. COAT-FV platelets may explain the unique role of -granule FV and the hemostatic effectiveness of young platelets.
Unique processing pathways within recipient antigen-presenting cells determine IgG immunity against donor platelet MHC antigens
Authors: Bang K. W. Annie , Speck Edwin R. , Blanchette Victor S. , Freedman John , Semple John W.
Published: October 2019

Recipient IgG immunity against leukoreduced donor platelets is dependent on indirect T-cell allorecognition and is suppressed in vivo by inhibitors (aminoguanidine, AMG) of inducible nitric oxide synthase (iNOS). To examine recipient processing pathways of donor platelet antigens, enriched macrophages (antigen-presenting cells [APC]) from BALB/c (H-2d) mice were pulsed with allogeneic C57BL/6 (H-2b) platelets and transfused weekly into naive BALB/c mice. Platelet-pulsed APC stimulated IgG antidonor antibody production in 45% of recipients by the second transfusion and in 100% by the sixth transfusion; this response was enhanced by pulsing in the presence of interferon-γ. By the sixth transfusion, high-titer IgG1 (mean titer 4990) and IgG2a (1933) isotypes specific for donor major histocompatibility complex (MHC) class I antigens were detected. Platelet pulsing in the presence of AMG or colchicine significantly inhibited the ability of APC to stimulate IgG alloantibodies; only 50% (P < .005) and 20% (P < .0001) of recipients, respectively, produced antibodies by the sixth transfusion. AMG inhibition was reversed by the addition of l-arginine, the substrate for iNOS. In contrast, pulsing in the presence of chloroquine, the proteasome inhibitory peptide MG115, or Brefeldin A enhanced APC immunity (70-100% of recipients antibody positive by the second transfusion [P < .05]); these agents allowed the pulsed APC to stimulate IgG2a but inhibited IgG1 production and this correlated with a reduction in serum interleukin (IL)-4 levels. The results suggest that for donor platelet antigens to stimulate IgG alloantibodies, recipient APC use the essential generation of nitric oxide and a noncytosolic, pH-independent processing pathway, which can be exploited as an effective immunotherapy target to further inhibit alloimmunization against leukoreduced platelets.
Elliptocytosis in patients with C-terminal domain mutations of protein 4.1 correlates with encoded messenger RNA levels rather than with alterations in primary protein structure
Authors: Morinière Madeleine , Ribeiro Leticia , Dalla Venezia Nicole , Deguillien Mireille , Maillet Philippe , Cynober Thérèse , Delhommeau François , Almeida Helena , Tamagnini Gabriel , Delaunay Jean , Baklouti Faouzi
Published: October 2019

Early biochemical studies defined 4 functional domains of the erythroid protein 4.1 (4.1R). From amino-terminal to carboxy-terminal, these are 30 kd, 16 kd, 10 kd, and 22/24 kd in size. Although the functional properties of both the 30-kd and the 10-kd domain have been demonstrated in red cells, no functional activities have been assigned to either the 16-kd or the 22/24-kd domain in these cells. We here describe new mutations in the sequence encoding the C-terminal 22/24-kd domain that are associated with hereditary elliptocytosis. An unusually mild phenotype observed in heterozygous and homozygous members of 1 family suggested heterogeneity in the pattern of expression of 4.1R deficiency. Using a variety of protein and messenger RNA (mRNA) quantification strategies, we showed that, regardless of the alteration in the C-terminal primary sequence, when the protein is produced, it assembles at the cell membrane. In addition, we found that alterations in red cell morphologic features and membrane function correlate with the amount of membrane-associated protein—and therefore with the amount of mRNA accumulated—rather than with the primary structure of the variant proteins. These data suggest that an intact sequence at exons 19 through 21 encoding part of the C-terminal 22/24-kd region is not required for proper protein 4.1R assembly in mature red cells.
Treatment of intractable autoimmune diseases in MRL/lpr mice using a new strategy for allogeneic bone marrow transplantation
Authors: Kushida Taketoshi , Inaba Muneo , Takeuchi Kenji , Sugiura Kikuya , Ogawa Ryokei , Ikehara Susumu
Published: October 2019

A new bone marrow transplantation (BMT) method for treating severe autoimmune diseases in chimeric resistant MRL/lpr mice is presented. The method consists of fractionated irradiation (5.5 Gy × 2), followed by portal venous (PV) injection of whole bone marrow cells (BMCs) from allogeneic normal C57BL/6 (B6) mice and intravenous (IV) injection of whole B6 BMCs 5 days after the PV injection (abbreviated as 5.5 Gy × 2 + PV + IV). All recipients survived more than 1 year after this treatment (more than 64 weeks after birth). Abnormal T cells (Thy1.2+/B220+/CD3+/CD4−/CD8−) present in MRL/lpr mice before the treatment disappear, and hematolymphoid cells are reconstituted with donor-derived cells. The treated mice are free from autoimmune diseases. Levels of autoantibodies (IgG/IgM anti-ssDNA antibodies and IgG/IgM rheumatoid factors) decrease to normal levels. Successful cooperation is achieved among T cells, B cells, and antigen-presenting cells (APCs) of the treated MRL/lpr mice when evaluated by in vitro anti-SRBC responses. Newly developed T cells are tolerant to both donor (B6)-type and host (MRL/lpr)-type major histocompatibility complex (MHC) determinants. These findings clearly indicate that severe autoimmune diseases in MRL/lpr mice are completely ameliorated by the treatment without recourse to immunosuppressants, and that the treated MRL/lpr mice show normal immune functions, strongly suggesting that this strategy would be applicable to humans.
Estrogen influences the differentiation, proliferation, and survival of early B-lineage precursors
Authors: Medina Kay L. , Strasser Andreas , Kincade Paul W.
Published: October 2019

B lymphocyte production in murine bone marrow is negatively regulated by sex steroids and the aim of this study was to identify early hormone sensitive checkpoints. Estrogen (E2) treatment reduced cμ+ pre-B cells, a change that occurred concomitantly with decreased Ig gene rearrangements and rag-1 transcripts. Estrogen decreased B lineage precursors in Ig transgenic mice, demonstrating that hormonal regulation is independent of the recombination process. B lineage precursors in Bcl-2 transgenic mice were resistant to estrogen treatment, suggesting that life/death decisions are involved in hormonal regulation. A previously uncharacterized population of CD43−cμ− B lineage precursors was identified in normal, Ig transgenic, and RAG−/− mice after estrogen treatment, revealing that down-regulation of CD43 can occur independent of Ig heavy chain expression. These cells expressed transcripts for both tdt andbcl-2, characteristics of early B-cell precursors. BrdU incorporation analysis revealed that the mitotic activity of early B-lineage cells is reduced in hormone-treated mice. We conclude that sex steroids modulate the production of B-lineage cells by influencing the differentiation, proliferation, and survival of early B-cell precursors. These findings are informative about mechanisms of hormonal regulation, as well as the significance of some differentiation-related events.
A recombinant bispecific single-chain antibody, CD19 × CD3, induces rapid and high lymphoma-directed cytotoxicity by unstimulated T lymphocytes
Authors: Löffler Anja , Kufer Peter , Lutterbüse Ralf , Zettl Florian , Daniel Peter T. , Schwenkenbecher Jan M. , Riethmüller Gert , Dörken Bernd , Bargou Ralf C.
Published: October 2019

Although bispecific antibodies directed against malignant lymphoma have been shown to be effective in vitro and in vivo, extended clinical trials so far have been hampered by the fact that conventional approaches to produce these antibodies suffer from low yields, ill-defined byproducts, or laborious purification procedures. To overcome this problem, we have generated a small, recombinant, lymphoma-directed, bispecific single-chain (bsc) antibody according to a novel technique recently described. The antibody consists of 2 different single-chain Fv fragments joined by a glycine-serine linker. One specificity is directed against the CD3 antigen of human T cells, and the other antigen-binding site engages the pan–B-cell marker CD19, uniformly expressed on the vast majority of B-cell malignancies. The construct was expressed in Chinese hamster ovary cells and purified by its C-terminal histioline tag. Specific binding to CD19 and CD3 was demonstrated by fluorescence-activated cell sorter analysis. By redirecting unstimulated primary human T cells derived from the peripheral blood against CD19-positive lymphoma cells, the bscCD19 × CD3 antibody showed significant cytotoxic activity at very low concentrations of 10 to 100 pg/mL and at effector to target cell ratios as low as 2:1. Moreover, strong lymphoma-directed cytotoxicity at low antibody concentrations was rapidly induced during 4 hours even in experiments without any T-cell prestimulation. Thus, this particular antibody proves to be much more efficacious than the bispecific antibodies described until now. Therefore, the described bscCD19 × CD3 molecule should be a suitable candidate to prove the therapeutic benefit of bispecific antibodies in the treatment of non-Hodgkin lymphoma.
Occurrence and distribution of antibiotic resistance genes in the water and sediments of Qingcaosha Reservoir, Shanghai, China
Authors: Huang Zhifeng , Zhao Wanting , Xu Ting , Zheng Binghui , Yin Daqiang
Published: October 2019

Background

Qingcaosha Reservoir is China’s largest river impoundment and an important drinking water resource for Shanghai city. Although antibiotics contamination in the reservoir has been reported, little is known about the presence of antibiotic resistance genes (ARGs). In this study, the occurrence and distribution of 12 ARGs and the class 1 integron gene (intI-1) in water and sediments collected from the reservoir were investigated for 1 year.

Results

The 12 ARGs were detected in both water and sediment samples, and no significant temporal or spatial variations were observed. Sulfonamide resistance genes were predominant in the reservoir with a detection frequency of 100%. Statistical analysis indicated a positive correlation in relative abundance between some ARGs such as sul1, sul2, and intI-1. Most ARGs were negatively correlated with total nitrogen and positively correlated with chemical oxygen demand and chlorophyll-a.

Conclusion

In general, ARGs were found to be prevalent in Qingcaosha Reservoir, among which sulfonamide resistance genes were the most dominant and tetB was the least. intI-1 may facilitate the proliferation and propagation of some ARGs, especially sul1 and sul2. The abundance of ARGs was well correlated with aquatic environmental factors in water, providing potential clues for the control of ARG contamination.
Characterization of LrgAB as a stationary phase-specific pyruvate uptake system in Streptococcus mutans
Authors: Ahn Sang-Joon , Deep Kamal , Turner Matthew E. , Ishkov Ivan , Waters Anthony , Hagen Stephen J. , Rice Kelly C.
Published: October 2019

Background

Our recent ‘-omics’ comparisons ofStreptococcus mutanswild-type andlrgAB-mutant revealed that this organism undergoes dynamic cellular changes in the face of multiple exogenous stresses, consequently affecting its comprehensive virulence traits. In this current study, we further demonstrate that LrgAB functions as aS. mutanspyruvate uptake system.

Results

S. mutansexcretes pyruvate during growth as an overflow metabolite, and appears to uptake this excreted pyruvate via LrgAB once the primary carbon source is exhausted. This utilization of excreted pyruvate was tightly regulated by glucose levels and stationary growth phaselrgABinduction. The degree oflrgABinduction was reduced by high extracellular levels of pyruvate, suggesting thatlrgABinduction is subject to negative feedback regulation, likely through the LytST TCS, which is required for expression oflrgAB. Stationary phaselrgABinduction was efficiently inhibited by low concentrations of 3FP, a toxic pyruvate analogue, without affecting cell growth, suggesting that accumulated pyruvate is sensed either directly or indirectly by LytS, subsequently triggeringlrgABexpression.S. mutansgrowth was inhibited by high concentrations of 3FP, implying that pyruvate uptake is necessary forS. mutansexponential phase growth and occurs in a Lrg-independent manner. Finally, we found that stationary phaselrgABinduction is modulated by hydrogen peroxide (H2O2) and by co-cultivation with H2O2-producingS. gordonii.

Conclusions

Pyruvate may provideS. mutanswith an alternative carbon source under limited growth conditions, as well as serving as a buffer against exogenous oxidative stress.Given the hypothesized role of LrgAB in cell death and lysis, these data also provide an important basis for how these processes are functionally and mechanically connected to key metabolic pathways such as pyruvate metabolism.
Prevalence and correlates of sexual intercourse among sexually active in-school adolescents: an analysis of five sub-Sahara African countries for the adolescent’s sexual health policy implications
Authors: Shayo Festo K. , Kalomo Mariam H.
Published: October 2019

Background

Early and unprotected sex with multiple partners among adolescents carries a high risk of acquiring HIV infections, other sexually transmitted infections as well as high rates of teenage pregnancy. Sub-Saharan Africa (SSA) has a higher burden of HIV/AIDS: the leading cause of deaths among adolescents. We estimated the prevalence and examined the correlates of sexual intercourse among in-school adolescents of SSA. The purpose is to inform the public health programs dedicated to tackling the burden of HIV/AIDS.

Methods

We did a secondary analysis of the Global School-based Student Health Surveys (GSHS) datasets pooled from five SSA countries Benin, Mozambique, Namibia, Seychelles, and Tanzania. Our current analysis included a sample of 15,318 in-school adolescents. The primary independent variables were ever had sexual intercourse and sex with multiple partners, while the dependent variables were smoking cigarettes, alcohol use, use of marijuana and amphetamine, and parental connectedness. We performed descriptive statistics, and multivariate logistic regression stratified by gender using SPSS Complex Sample Statistics. A p-value of less than 0.05 was considered statistically significant at 95% confidence intervals.

Results

Out of 15,318 participants, the overall prevalence of ever had sexual intercourse and sex with multiple partners were 43·5% (6670) and 20·9% (3204), respectively. In overall and across each country, male adolescents had a significantly higher proportion of sex with multiple partners than female adolescents, p < 0·001. The predictors of sexual intercourse with multiple partners in both male and females were smoking cigarettes, alcohol use, and use of marijuana and amphetamine. Female adolescents who smoked cigarettes and used marijuana had a significant likelihood of sex with multiple partners than male adolescents: [aOR 3.6, 95% CI: 2.6-5.1] vs [aOR 2.1, 95% CI: 1.7-2.7] and [aOR 2.4, 95% CI: 1.6-3.7] vs [aOR 1.9, 95% CI: 1·3-2·7], respectively.

Conclusions

Adolescents sexual intercourse and more especially sex with multiple partners was prevalent and strongly correlated with substance use. However, the correlation was higher among female adolescents than male adolescents. A customized public health intervention that targets multiple risk factors concurrently may benefit adolescents with clustering of sexual and non-sexual risk-taking behaviors.
Is sitting invisible? Exploring how people mentally represent sitting
Authors: Gardner Benjamin , Flint Stuart , Rebar Amanda L. , Dewitt Stephen , Quail Sahana K. , Whall Helen , Smith Lee
Published: October 2019

Background

Growing evidence suggests that prolonged uninterrupted sitting can be detrimental to health. Much sedentary behaviour research is reliant on self-reports of sitting time, and sitting-reduction interventions often focus on reducing motivation to sit. These approaches assume that people are consciously aware of their sitting time. Drawing on Action Identification Theory, this paper argues that people rarely identify the act of sitting as ‘sitting’ per se, and instead view it as an incidental component of more meaningful and purposeful typically-seated activities.

Methods

Studies 1 and 2 explored whether people mentioned sitting in written descriptions of actions. Studies 3–5 compared preferences for labelling a typically desk-based activity as ‘sitting’ versus alternative action identities. Studies 6 and 7 used card-sort tasks to indirectly assess the prioritisation of ‘sitting’ relative to other action descriptions when identifying similar actions.

Results

Participants rarely spontaneously mentioned sitting when describing actions (Studies 1–2), and when assigning action labels to a seated activity, tended to offer descriptions based on higher-order goals and consequences of action, rather than sitting or other procedural elements (Studies 3–5). Participants primarily identified similarities in actions based not on sitting, but on activities performed while seated (e.g. reading; Studies 6–7).

Conclusion

‘Sitting’ is a less accessible cognitive representation of seated activities than are representations based on the purpose and implications of seated action. Findings suggest that self-report measures should focus on time spent in seated activities, rather than attempting to measure sitting time via direct recall. From an intervention perspective, findings speak to the importance of targeting behaviours that entail sitting, and of raising awareness of sitting as a potential precursor to attempting to reduce sitting time.
Crosslinked sulfonated polyacrylamide (Cross-PAA-SO3H) tethered to nano-Fe3O4 as a superior catalyst for the synthesis of 1,3-thiazoles
Authors: Shahbazi-Alavi Hossein , Khojasteh-Khosro Sheida , Safaei-Ghomi Javad , Tavazo Maryam
Published: October 2019

Crosslinked sulfonated polyacrylamide (Cross-PAA-SO3H) attached to nano-Fe3O4 as a superior catalyst has been used for the synthesis of 3-alkyl-4-phenyl-1,3-thiazole-2(3H)-thione derivatives through a three-component reactions of phenacyl bromide or 4-methoxyphenacyl bromide, carbon disulfide and primary amine under reflux condition in ethanol. A proper, atom-economical, straightforward one-pot multicomponent synthetic route for the synthesis of 1,3-thiazoles in good yields has been devised using crosslinked sulfonated polyacrylamide (Cross-PAA-SO3H) tethered to nano-Fe3O4. The catalyst has been characterized by Fourier-transform infrared spectroscopy (FT-IR), scanning electron microscope (SEM), dynamic light scattering (DLS), X-ray powder diffraction (XRD), energy-dispersive X-ray spectroscopy (EDS), thermogravimetric analysis (TGA) and vibrating-sample magnetometer (VSM).
Estimating the productive potential of five natural forest types in northeastern China
Authors: Wu Zhaofei , Zhang Zhonghui , Wang Juan
Published: October 2019

Background

There is a serious lack of experience regarding the productive potential of the natural forests in northeastern China, which severely limits the development of sustainable forest management strategies for this most important forest region in China. Accordingly, the objective of this study is to develop a first comprehensive system for estimating the wood production for the five dominant forest types.

Methods

Based on a network of 384 field plots and using the state-space approach, we develop a system of dynamic stand models, for each of the five main forest types. Four models were developed and evaluated, including a base model and three extended models which include the effects of dominant height and climate variables. The four models were fitted, and their predictive strengths were tested, using the “seemingly unrelated regression” (SUR) technique.

Results

All three of the extended models increased the accuracy of the predictions at varying degrees for the five major natural forest types of northeastern China. The inclusion of dominant height and two climate factors (precipitation and temperature) in the base model resulted in the best performance for all the forest types. On average, the root mean square values were reduced by 13.0% when compared with the base model.

Conclusion

Both dominant height and climate factors were important variables in estimating forest production. This study not only presents a new method for estimating forest production for a large region, but also explains regional differences in the effect of site productivity and climate.
Spinocerebellar Ataxia type 29 in a family of Māori descent
Authors: Ngo Kathie J. , Poke Gemma , Neas Katherine , Fogel Brent L.
Published: October 2019

Background

Mutations in the Inositol 1,4,5-Trisphosphate Receptor Type 1 (ITPR1) gene cause spinocerebellar ataxia type 29 (SCA29), a rare congenital-onset autosomal dominant non-progressive cerebellar ataxia. The Māori, indigenous to New Zealand, are an understudied population for genetic ataxias.

Case presentation

We investigated the genetic origins of spinocerebellar ataxia in a family of Māori descent consisting of two affected sisters and their unaffected parents. Whole exome sequencing identified a pathogenic variant, p.Thr267Met, in ITPR1 in both sisters, establishing their diagnosis as SCA29.

Conclusions

We report the identification of a family of Māori descent with a mutation causing SCA29, extending the worldwide scope of this disease. Although this mutation has occurred de novo in other populations, suggesting a mutational hotspot, the children in this family inherited it from their unaffected mother who was germline mosaic.
In memory to Professor D. Ya. Shuryghin
Authors: Journal Staff Problems of Endocrinology
Published: October 2019

The article is devoted to the memorable date, the 70th anniversary of the birth of Professor Dorofey Yakovlevich Shurygin, one of the prominent endocrinologists of the country, honored scientist of the RSFSR.
A second KIGS communication on side effects of growth hormone therapy
Authors: Chatlein P. J. , Wilton P.
Published: October 2019

By March 25, 1992, among the 16,604 patients included in the KIGS database who had been treated with growth hormone (GH) for at least one year, 135 had serious side effects (SE). Among the most frequent SE are relapses of craniopharyngioma (17) and other brain tumors (11). Changes in bone tissue, joints and muscles were more rarely observed. 7 patients had convulsive syndrome. More information is needed to assess the effect of GH therapy on these complications.
Biosynthetic insulin effect on red cell membrane lipid peroxidation in patients with insulin-dependent diabetes
Authors: Balashova T. S. , Golega Ye. N. , Rudko I. A. , Balabolkin M. I. , Kubatiev A. A.
Published: October 2019

The aim of our research was elucidation of a relationship between red cell membrane lipid peroxidation (LPO) and antioxidant' defense enzymes, on the one hand and the age disease duration and presence of vascular complications in patients with type I diabetes mellitus on the other. The possibility of correcting red cell peroxide status with human insulin preparations was investigated. Red cell membrane LPO was found increased more than twofold and antioxidant defense enzymes activities virtually unchanged vs. controls in 16 patients with diabetes aged 20 to 43. These characteristics of red cell peroxidation status do not depend on patients age disease standing or presence of vascular complications. A twelve-week therapy with biosynthetic insulin resulted in complete normalization of LPO processes in patients with angiopathies aged under 35 and with disease standing of less than 10 years. In diabetics with angiopathies aged over 35 and disease standing of more than 10 years red cell MDA level reduced under the effect of therapy with human insulin preparations but was still increased vs. that in healthy donors by 1.5 times. Red cell GP and SOD activities reduced in the course of insulin therapy in all the examined groups of diabetics. Catalase activity increased by approximately 50% in patients with angiopathies those aged over 35 and a disease standing of more than 10 years under the effect of insulin. In the rest groups of patients catalase activity did not differ from its initial level. Our results permit us recommending besides human insulin preparations antioxidant therapy for patients with vascular complications those aged over 35 and a disease standing of more than 10 years.