bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinical Outcomes in Patients Receiving Venous Thromboembolism (VTE) Prophylaxis after Orthopedic Surgery (OS). A Systematic Review and Meta-Analysis of Proportions.
Authors: Lazo-Langner Alejandro , Coyle Douglas , Barrowman Nicholas J. , Ramsay Tim , Wells Philip S. , Scarvelis Dimitrios , Forgie Melissa A. , Rodger Marc 
Published: April 2020

Abstract
Major VTE is the most frequent complication of OS. Current recommendations are to administer prophylaxis with an anticoagulant agent for at least 7–10 days. Numerous studies have evaluated different agents for this purpose. Although the most recent studies are usually methodologically sound, several studies have been hampered by inappropriate designs and insufficient sample sizes. In order to help with the design of future trials we conducted a systematic review of randomized trials evaluating short-term (&lt; 15 days) administration of anticoagulants for VTE prophylaxis in OS and performed a MA of simple proportions to estimate the overall incidence of major VTE (proximal VTE, pulmonary embolism (PE), or death from PE), total VTE (proximal and distal VTE, PE or death from PE), and major bleeding episodes (as defined by the authors and defined using a definition similar to the one proposed by the International Society on Thrombosis and Haemostasis-ISTH). We included randomized trials comparing different drugs for VTE prophylaxis in OS (hip and knee arthroplasty and hip fracture surgery) using systematic evaluation of VTE (ultrasound or venography, pulmonary angiography, tomographic angiography, or ventilation perfusion lung scan). Heterogeneity of proportions was evaluated using a chi ² test and pooled estimates of proportions were obtained using a fixed or a random effects model as appropriate. In the latter the weights were estimated as proposed by Laird and Mosteller. We retrieved 55 studies (135 research arms) which enrolled 42,131 patients. The percentage and variance of major and total VTE and major bleeding are shown in table 1. The total number of events and the number of evaluable patients are shown in table 2.

We found differences in the percentage of clinical outcomes associated with the use of different agents for VTE prophylaxis after OS, however, because of the analytical strategy used no estimation of odds or risk reduction can be derived from this data. We believe that these estimates will be of help for the design of future studies.
A Novel Cytokine, FPT025, Regulates Myeloid Growth and Differentiation Via the M-CSF Receptor.
Authors: Lin Haishan , Huang Min Mei , Leo Cindy , Ji May , Wu Ge , Zhou Aileen , Halenbeck Robert , Qin Minmin , Linnemann Thomas , Behrens Dirk , Giese Scott , Bosch Elizabeth , Justin Katrina , Pham Amber , Lee Ernestine , Hestir Kevin , Zhang Hongbing , Wong Justin , Wang Yan , Chu Keting , Doberstein Steve , Williams Lewis T. , Hollenbaugh Diane 
Published: April 2020

Abstract
To identify novel protein therapeutics we have utilized an integrated screening system in which a collection of full-length human cDNA clones, encoding virtually all secreted proteins and receptors, was assembled. Soluble secreted proteins were expressed in a high-throughput format using human cells as the expression host and tested in high-throughput cell-based assays. Through this approach, a novel cytokine, FPT025, was identified in a human monocyte proliferation assay. FPT025 has no apparent sequence homology to known cytokines or any other genes and is expressed in human spleen, skin, brain, and other tissues. The purified recombinant FPT025 protein stimulated human primary monocyte proliferation and/or survival. FPT025 protein specifically bound to human primary monocytes and activated ERK1/2 phosphorylation in primary monocytes as well as in a human monocytic cell line, THP-1. FPT025 promoted formation of the myeloid lineage colonies, CFU-M, in a human bone marrow colony formation assay and enhanced proliferation of cells with myeloid cell surface markers from human monocytes. To identify the receptor of FPT025, a collection of extracellular domains (ECD) of transmembrane proteins was screened for their ability to block FPT025 activation of monocyte proliferation. In this screen, we found that FPT025 binds to the M-CSF receptor (M-CSFR) with high affinity. Furthermore, we showed that the binding of FPT025 to M-CSFR was specific and could be competed by M-CSF. The soluble ECD of M-CSFR inhibited both the binding of FPT025 to M-CSFR and the activity of FPT025 in monocyte proliferation. Therefore, FPT025 functions as a novel ligand of the M-CSF receptor and participates in the regulation of myeloid lineage differentiation, proliferation, and survival.
Morpholino Knockdown of Jak2a Inhibits Primitive and Definitive Hematopoiesis in Zebrafish.
Authors: Lin Hui-feng , Zhang Jing , Handin Robert I. 
Published: April 2020

Abstract
In zebrafish, the primitive wave of hematopoiesis is specified at 3 somites and is marked by two lateral stripes, which express early hematopoietic genes. The coalescence of these stripes forms the intermediate cell mass (ICM). This phase of hematopoiesis lasts about 4 days with erythrocytes entering the circulation 26 hpf. Between 36 and 42 hpf, the definitive wave of hematopoiesis commences. It is marked by the presence of hematopoietic progenitor cells in the dorsal aortic wall. Later in day 3 of development, hematopoietic stem cells from the dorsal aorta migrate into the interstitium of the mesonephros, which becomes the major hematopoietic organ in adult fish. Jak2 is a member of the Janus kinase family of non-receptor tyrosine kinases which plays an important role in hematopoiesis by transmitting cytokine-induced signals to associated STATs (signal transcription and activation transducers). The phosphorylated STATS migrate to the nucleus and induce gene transcription. The knockout of Jak2 in mice is embryonic lethal with a severe defect in erythropoiesis. We and others have observed that Jak2 is duplicated in zebrafish with one of the duplicated genes, jak2a, expressed exclusively in hematopoietic tissues. We have knocked down jak2a with anti-sense morpholinos to assess its role in hematopoiesis. O-dianisidine staining of embryos 2dpf shows a reduction in the primitive wave of erythropoiesis in the jak2a morphants. Using the CD41-GFP transgenic fish line we observed markedly reduced and, in some cases, the complete absence of circulating thrombocytes in jak2a morphants. To help understand how jak2a knockdown impairs hematopoiesis we examined the expression of a panel of early hematopoietic genes in the ICM of 20 to 24 hpf fish by whole mount in situ hybridization (WISH). There was an overall reduction in the expression of lmo2, scl, gata-1 and gata-2. Upon closer observation it was apparent that the reduction was due to the loss of certain cells from the ICM with normal staining intensity in the remaining cells. We also observed reduced cyclin D1 expression and elevated caspase 3 activity, which strongly suggested programmed cell death was activated in the jak2a morphant. We believe that there is a selective loss due to cell cycle arrest of the more mature hematopoietic cells, which require the expression of jak2a for their survival with retention of early hematopoietic progenitors and hematopoietic stem cells in the ICM. These observations in developing zebrafish embryos establish the important role of jak2a in hematopoiesis and provide new insights into the stage of hematopoiesis where jak2a is critically important.
Tumor Cell-Derived HLA-B-Associated Transkript 3 (BAT3) Is a Ligand for NKp30 and Activates NK Cells.
Authors: von Strandmann Elke Pogge , Simhadri Venkateswara R. , von Tresckow Bastian , Sasse Stefanie , Reiners Katrin S. , Hansen Hinrich P. , Rothe Achim , Borchmann Peter , McKinnon Peter J. , Hallek Michael , Engert Andreas 
Published: April 2020

Abstract
Natural-killer (NK)-cells are lymphocytes of the innate immune system, which directly attack tumor and virus-infected cells. They provide a link between innate and adaptive immunity through crosstalk with dendritic cells (DCs) and mediate T-cell activation. Among the activating and inhibitory receptors that regulate NK cell activity, the orphan NKp30 receptor (NCR3, CD337) plays a special role as NKp30 does not only induce target cell lysis but is also crucial for the interaction between NK cells and dendritic cells. However, so far the cellular ligands for NKp30 have remained elusive.
Using a yeast two hybrid approach with the extracellular NKp30 sequence as bait we were able to isolate a putative NKp30 ligand from a tumor cDNA library. Sequence analysis revealed that the cDNA encoded for BAT3 (HLA-B-associated transcript 3) which originally had been cloned from the human major histocompatibility complex by chromosome walking and mapped to chromosome 6p21.3 within the HLA complex. BAT3 was described as a nuclear protein characterized by an N-terminal ubiquitin-like region, a polyproline stretch and a highly conserved BAG-(Bcl-2-associated athanogene) domain. Until now, the function of BAT3 in mammals is not clearly defined. With BAT3-specific antibodies and by means of laser scanner microscopy and Western Blotting we demonstrate that tumor cell lines and immature as well as mature DCs express BAT3. Upon co-cultivation with NK cells or exposure to stress signals such as heat shock, we observed that BAT3 translocates from the nucleus to the cell-surface and the protein is released from tumor cells into the extracellular environment. BAT3 binds directly and specifically to NKp30 on NK cells and triggers NK cell-mediated cytokine release and NKp30-dependent cytotoxicity. The inhibition of endogenous BAT3 using polyclonal antibodies prevents tumor rejection in vivo, demonstrating that BAT3 is necessary for the tumor rejection in a xenograft tumor model.
Thus, BAT3 is the long sought cellular NKp30 ligand and exhibits a novel mechanism by which a nuclear protein activates NK cells via NKp30-engagement after its release to the cell-surface and the cell environment. The isolation of the first cellular NKp30-ligand provides the basis for a better molecular understanding of NK cell-regulation and allows the development of clinical applications targeting NKp30 for the immunotherapy.
The Unexpected Role of C3a-C3aR Axis in Maturation of Erythroid Cells - Implications for Pathogenesis of Hypoxia-Related Polycythemias.
Authors: Ratajczak Janina , Reca Ryan , Machalinski Bogdan , Maciejewski Jaroslaw , Laughlin Mary , Ratajczak Mariusz Z. 
Published: April 2020

Abstract
Hypoxia leads both to secondary polycythemia and activation of the complement (C) cascade in bone marrow (BM). We reported that C is activated in BM during hypoxia by the IgM-dependent classical pathway (Blood2006, 107:835–840). These observations prompted us to study the potential role of C cleavage fragments in erythropoiesis. We noticed that human and murine cells from the erythroid lineage express functional G-protein coupled C3aR and that C3 cleavage fragment - anaphylatoxin C3a enhances SDF-1-mediated chemotaxis of erythroid progenitors and induces in normal erythroblasts i) adhesion to fibronectin, ii) calcium flux and iii) phosphorylation of MAPKp42/44. To further explore a potential role of the C3a-C3aR axis in erythropoiesis we focused on C3aR−/ − mice. We noticed that these animals as compared to wild type (wt) littermates have i) lower hematocrits and ii) display delayed recovery of hematocrit values after sublethal irradiation. More importantly, we noticed that the number of clonogeneic BFU-E progenitors in BM of C3aR−/ − mice, but not CFU-GM and CFU-Meg, is reduced by ~65%. Thus, in order to learn more about the role of the C3a-C3aR axis in erythropoiesis we moved to an in vitro model and costimulated normal human CD34+ or murine SKL cells with C3a or desArgC3a in the presence of suboptimal doses of EpO and KL, but no effect on BFU-E colony formation was observed. We found, however, that C3a increases in early erythroid cells expression of mRNA for VEGF and MMP-2 - which are important for maturation/enucleation and egress of erythrocytes from the BM into peripheral blood. These data were subsequently confirmed by ELISA (VEGF) and zymography (MMP-2). Furthermore, we noticed that at the morphological level C3aR−/ − mice as compared to wt littermates had significantly reduced number of erytropoietic islands in BM. This suggests that in the absence of the C3aR, maturing erythroblasts do not traffic properly in the marrow microenvironment in BM and do not associate with C3 producing central macrophages in erythropoietic islands. In conclusion the C3a-C3aR axis modulates the interaction of erythroid progenitors with the hematopoietic environment and we conclude that it plays an important and underappreciated role in developmental trafficking of erythroid cells in BM. Thus C3a may enhance the erythropoietic effects of erythropoietin in polycythemias related to hypoxia. Furthermore, since hypoxia activates C, we postulate that C cleavage fragments could be involved in the pathogenesis of hypoxia-related polycythemias and C3aR antagonists could find a potential supportive role in controlling erythropoiesis in these patients.
Epo Receptor Circuits to Irs2, Foxo3a, Trb3 and Bim1 Expression, and Erythroblast Survival.
Authors: Bogacheva Olga , Bogachev Oleg , Sathyanarayana Pradeep , Menon Madhu P. , Kapelle William S. , Houde Estelle , Wojchowski Don M. 
Published: April 2020

Abstract
Epo cytoprotection of developing erythroblasts mechanistically is thought to involve 1] PI3K and Akt pathways (eg. mTOR, Foxo3A inhibition) as engaged via a distal EpoR PY479 site, and 2] anti-apoptotic effects as exerted by EpoR proximal box1 and PY343 signals (eg. Bcl-xl transcription). To globally assess Epo-regulation of survival- associated genes, a maximally Epo-responsive cohort of bone-marrow derived KitposCD71high erythroblasts was prepared, purified and used in Affymetrix 430–2.0 array based profiling of Epo responsive genes. Quadruplicate array analyses and RT-QPCR revealed selective modulation of PI3K pathway associated factors Irs2, p85-alpha, Trb3, and Foxo3a as well as the Bcl2-related factor Bim1. Irs2, p85-alpha and Trb3 were induced three- to five-fold, while Foxo3a was rapidly down-modulated. As studied using knocked-in minimal EpoR alleles, induction of Irs2 and Trb3 (a pseudo-kinase Akt inhibitor) depended sharply upon an EpoR PY343 Stat5 binding site. Epo down-modulation of Foxo3a (as well as Trb2 and Bim1), in contrast, occurred via EpoR PY-independent routes. Unexpectedly, the EpoR allele, EpoR-H-Y343 also protected erythroblasts from LY294002 and wortmanin induced apoptosis, while an EpoR-HM-F343 allele was hyper-sensitive. For EpoRH, this proved to correlate specifically with Y343 dependent phosphorylation of GAB2. PY343 couples specifically to Stat5, and phospho-Stat5 therefore is proposed to recruit GAB2 and enable its PY452 phosphorylation. GAB2 then provides a shunt to PI3K activation. Findings provide new insight into Epo-regulation of a select set of PI3K-associated genes, and core EpoR cytoprotective mechanisms.
Tumor Cell-Associated Tissue Factor Supports Metastatic Potential through Both NK Cell-Dependent and -Independent Mechanisms.
Authors: Palumbo Joseph , Barney Kelley A. , Flick Matthew J. , Talmage Kathryn E. , Kombrinck Keith W. , Hu Zhiwei , La Jeunesse Christine M. , Degen Jay L. 
Published: April 2020

Abstract
Multiple lines of evidence indicate that the hemostatic system contributes to cancer dissemination. Previous studies have shown that tumor cell-associated tissue factor (TF) expression is a crucial determinant of metastatic potential. Furthermore, we have shown that tumor cell-associated TF supports metastatic potential through mechanism(s) dependent on multiple circulating hemostatic system components, including prothrombin, platelets, fibrinogen and, more recently, fXIII. At least two of these circulating hemostatic factors (platelets and fibrinogen) have been shown to support metastatic potential by impeding the clearance of recently embolized tumor cells by natural killer (NK) cells. It is reasonable to hypothesize that tumor cell-associated TF expression also supports metastatic potential by a mechanism coupled to NK cell function. Here, we used C57BL/6-derived, Ras-transformed tumor cell lines expressing wildtype murine tissue factor (TFWT), a mutant TF lacking the intracytoplasmic portion (TFΔTail), or no tissue factor (TFO) to directly examine the interplay between tumor cell-associated TF and NK cell function in determining metastatic potential. Each of these cell lines was capable of robust, comparable tumor growth in wildtype C57BL/6 mice. Loss of either platelet function or fibrinogen significantly diminished the metastatic potential of TFWT cells, but this effect was entirely abrogated by the concomitant loss of NK cells. Similar results were obtained with TFΔTail cells, indicating that the cytoplasmic portion of TF is not critical to these interactions. To determine if the increase in metastatic potential conferred by tumor cell-associated TF expression is entirely linked to NK cell function, we compared the metastatic potential and early survival of TFWT and TFO cells in mice with and without NK cells. TFOcells rarely formed any visible metastatic foci in mice with intact NK cell function, while TFWT cells were aggressively metastatic. Importantly, TF expression remained a significant determinant of metastatic potential even in mice lacking NK cells. Comparisons of the early fate of TFWT and TFO cells revealed that TF expression was not a determinant of initial tumor cell localization within the lungs. Rather, TF expression supported the sustained adherence and/or survival of tumor cells. Taken together, these data indicate that one mechanism linking tumor cell-associated TF expression to metastatic potential is coupled to circulating hemostatic factors and results in impaired NK cell-mediated clearance of recently established micrometastatic foci. However, TF expression also supports metastasis by at least one additional mechanism that is independent of natural killer cell function.
Clinical Significance of JAK2V617F Homozygosity in the Chronic Myeloproliferative Disorders. A Study on 1306 Patients.
Authors: Vannucchi Alessandro M. , Barosi Giovanni , Rambaldi Alessandro , Marchioli Roberto , Barbui Tiziano ,   for the GIMEMA MPD, WP
Published: April 2020

Abstract
The JAK2V617F mutation is variably found in the Ph’-negative chronic myeloproliferative disorders (MPD); in about 25% of polycythemia vera (PV) or idiopathic myelofibrosis (IM) and in 1–3% of essential thrombocythemia (ET) patients (pts), mitotic recombination causes the mutation to be harboured in the homozygous status. While it has been suggested that homozygosity influences some cellular functions, its clinical significance has not been elucidated yet due to the low number of homozygotes in published series. To this end, a retrospective, multicentric study, that enrolled 1306 pts, was designed within the Myeloproliferative Working Group of the Italian GIMEMA association; all pts who had been typed for JAK2 mutation using the BsaXI digestion assay (Baxter et al, Lancet, 2005) were included. There were 397 PV (30.4%), 639 ET (48.9%), 270 IM (20.7%) pts. JAK2V617F was found in 878 (67.3%), accounting for 82%, 60%, and 64%, respectively, of PV, ET, or IM pts. The homozygotes (Ho) were 183 (20.8%), with a frequency of 26.3% in PV, 2.2% in ET, and 24.1% in IM. Ho pts, irrespectively of the underlying diagnosis, presented higher leukocyte count and hematocrit, while platelet count was lower, as compared to heterozygous (He) or wild-type (WT). The frequency of splenomegaly rose from 26% to 41% to 52% in WT, He and Ho PV pts; from 19% to 22% to 62% in ET; and from 61% to 67% to 89% in IM. Acquagenic pruritus was referred by 12%, 28% and 42% of WT, He, and Ho PV pts, while the corresponding figures were 4%, 12%, and 26% in IM; no difference was noted among ET pts. Also the frequency of systemic symptoms was significantly higher among Ho pts in all three disease groups. 195 pts (14.9% of total) presented major thromboses at diagnosis (71 PV, 114 ET, 10 IM, accounting for 18%, 18%, and 4% in each group, respectively), while 144 events were recorded in the follow-up (15%, 12%, 4% in PV, ET, or IM). The incidence of thrombosis at diagnosis was 12%, 21% and 15% in WT, He or Ho PV pts, while the corresponding figures were 11%, 22% and 50% in ET, and 1%, 7% and 3% in IM. The relative risk (RR) for major thrombosis after diagnosis (adjusted for gender, age, and previous thrombosis) was significantly higher in Ho than in He pts both in ET (RR 4.6 and 1.5, respectively, by taking as 1 the WT pts) and IM (RR 3.2 and 1.6) pts, unlike in PV (RR 0.8 and 0.9, respectively). Ho pts evolved more frequently into myeloid metaplasia than He or WT in both PV (12% vs 0% and 1%, respectively) and ET (14% vs 5%, and 2%). These data support the contention that the JAK2V617F homozygous mutational status associates with a more symptomatic disease across the MPD clinical entities, and identify the unfrequent Ho ET patients as a specific subgroup with a particularly aggressive disease.
Highly Sensitive Real-Time PCR of V617F-JAK2-Mutation To Monitor Minimal Residual Disease and Guide Donor Lymphocte Infusion after Allogeneic Stem Cell Transplantation in Patients with Myelofibrosis.
Authors: Kroeger Nicolaus , Badbaran Anita , Holler Ernst , Hahn Joachim , Kobbe Guido , Bornhauser Martin , Reiter Andreas , Zabelina Tatjana , Zander Axel , Fehse Boris 
Published: April 2020

Abstract
The V617F-JAK2-mutation occurs in about 50 % of patients with myelofibrosis and is a reliable marker to monitor residual disease after allogeneic stem cell transplantation. The establishment of valid complete remission criteria for myelofibrosis especially after allogeneic stem cell transplantation remains a major issue. We developed a new, highly sensitive real-time PCR to monitor and quantify V617F-JAK2-positive cells after dose-reduced allogeneic stem cell transplantation. The mutated differs from the wild-type JAK2 allele by just one nucleotide exchange (G à T) leading to the valine to phenylalanine (V à F) transition. Using PrimerExpress® we designed a TaqMan® PCR where the reverse primer (RP) terminates at the (3′) nucleotide corresponding to this point mutation. Thus, this reverse primer should bind with higher affinity to the mutated than to the wild-type allele. To increase the specificity while conserving optimal sensitivity of the MRD-specific PCR we generated a set of primers shortened each time by one nucleotide at their 5’ end. In parallel, all those shortened primers were designed to contain an additional mutation at the third to last 3’ position. This allowed us to identify the reverse primer combining high specificity with so far not reported sensitivity(0.01 %). After 22 allogeneic stem cell transplantation procedures in 21 JAK2-positive patients with myelofibrosis, 78 % became PCR-negative. In 15 out of 17 patients (88 %), JAK2 remained negative after a median follow-up of 20 months. JAK2-negativity was achieved after a median of 89 days post allograft (range, 19 – 750 days). A significant inverse correlation was seen for JAK2 positivity and donor cell chimerism (r: −0.91, p&lt;0.001). Four of five patients who never achieved JAK2-negativity fulfilled during the entire follow-up all criteria for complete remission recently proposed by the International Working Group, suggesting a major role for JAK2 measurement to determine depths of remission. In one case, residual JAK2 positive cells could be eliminated by donor lymphocyte infusion, supporting the graft versus myelofibrosis effect. In conclusion, allogeneic stem cell transplantation after dose-reduced conditioning induces high rates of molecular remission in JAK2-positive myelofibrosis-patients and quantification. V617F-JAK2 mutation by real-time PCR allows detecting minimal residual disease to guide adoptive immunotherapy.