bluuebunny/crossref_metadata_2025_split huggingface dataset:
Polymorphism of the α2A-Adrenergic Receptor Gene and Epinephrine-Induced Platelet Aggregation.
Authors: Afshar-Kharghan Vahid , Sun Carrol , Zhang Hui , Wood Jennifer , Leal Suzanne , Dong Jing-Fei , Bray Paul F. 
Published: October 2019

Abstract
The α2A-adrenergic receptor mediates important physiologic responses, such as regulation of blood pressure, control of sympathetic autonomic responses, as well as the platelet response to epinephrine. The gene encoding the α2A-adrenergic receptor is polymorphic. One of the polymorphisms, known as the Dra I polymorphism, is located in the 3′ untranslated region of this gene. The Dra I polymorphism of the α2A-adrenergic receptor is associated with a higher incidence of systemic arterial hypertension among African-Americans. Although the presence of this polymorphism was known by Southern-blot analysis of the human genomic DNA for more than a decade, the exact location and nature of this polymorphism was unknown. We have located the Dra I polymorphism in the 3′ untranslated region of the α2A-adrenergic receptor gene (G1838A) and developed a PCR-based detection method. The allele and haplotype frequencies for polymorphisms of the α2A-adrenergic receptor gene in different ethnic groups were identified. After studying the correlation between epinephrine-induced platelet aggregation and the genotypes for the G1838A polymorphism among 235 human subjects, it was found that the 1838G allele is associated with an increase in the level of platelet aggregation in response to epinephrine among African-Americans, but not among Caucasians or Hispanics. This polymorphism may be responsible for the variability in the epinephrine-induced platelet aggregation among individuals and acts as a risk factor for thrombotic vascular event.
Transcriptional Regulation of the SCL Locus: Identification of an Enhancer That Targets Primitive Erythroid Cells.
Authors: Delabesse* Eric , Ogilvy* Sarah , Chapman Michael A. , Göttgens Berthold , Green Anthony R. 
Published: October 2019

Abstract
The stem cell leukaemia (SCL) gene encodes a bHLH protein that is essential for the formation of all haematopoietic lineages. In addition, maintenance of SCL expression is required for normal differentiation along the erythroid and megakaryocytic lineages, whereas failure to downregulate SCL transcription during T-cell differentiation is associated with T-cell ALL. Current evidence therefore demonstrates that appropriate transcriptional regulation is essential for the biological functions of SCL and this focuses attention on the mechanisms whereby transcription of SCL itself is initiated and maintained.
We have previously used biochemical, comparative genomic and transgenic assays to identify 5 distinct enhancers which target different subdomains of the normal SCL expression pattern. However these enhancers do not explain how erythroid expression of SCL is achieved and we have postulated the existence of an additional erythroid enhancer. It is also unclear whether the known SCL enhancers regulate neighbouring genes within the SCL locus. We have therefore quantitated the transcripts from SCL and its neighbouring genes in a large panel of human and murine haematopoietic cell types. Our results reveal a striking and unexpected co-expression of SCL and its downstream neighbour MAP17 (r=0.8; n=31). We demonstrate the existence of appropriately spliced low abundance SCL-MAP17 fusion transcripts suggesting that co-expression reflects transcriptional read-through rather than a shared enhancer. A systematic survey of histone H3 and H4 acetylation throughout the SCL locus was also performed in both cell lines and primary haematopoietic cells. A peak of acetylation downstream of MAP17 (and 40 kb downstream of SCL exon 1a) was found to correlate with expression of SCL but not other neighbouring genes. This region contains peaks of homology in 4-way genomic sequence comparisons (human/dog/mouse/rat) and functions as an erythroid-restricted enhancer in vitro. Morever, in transgenic mice this enhancer directs b-galactosidase expression to the vast majority of circulating primitive erythroblasts but not to fetal liver definitive erythroblasts.
Recruitment of mSin3A and Histone Deacetylase 2 (HDAC2) by the Chromatin Remodeling Protein BRG1 Mediates Transcriptional Repression in Erythroid Progenitors.
Authors: Xu Zhixiong , Meng Xianzhang , Cai Ying , Brandt Stephen J. 
Published: October 2019

Abstract
The Swi/Snf chromatin remodeling complexes are critical for activating or repressing transcription of specific genes and require one of two homologous ATPases, Brg1 and Brm. Although Brg1 is also known to be involved in ß-globin gene transcription and a recent report showed that Brg1-containing complexes could be recruited to the ß-globin promoter and its locus control region (LCR), the exact roles of Brg1 in erythroid gene expression and differentiation have not been directly investigated. We previously showed that Brg1, but not Brm, is recruited to the Protein 4.2 (P4.2) promoter by a multi-protein DNA-binding complex containing TAL1/SCL, E47, GATA-1, LMO2, and Ldb1. This complex transactivates P4.2 gene expression through tandem E box-GATA elements in its proximal promoter. We report here that overexpression of Brg1 in murine erythroleukemia (MEL) cells significantly reduced the proportion of benzidine-stained cells and dramatically inhibited expression of mRNA for P4.2 and ß-globin but not glycophorin A and erythroid Kruppel-like factor in MEL cells induced to differentiate with dimethyl sulfoxide (DMSO). Further, overexpression of Brg1 repressed P4.2 promoter activity in a dose-dependent manner in DMSO-treated MEL cells, while treatment of cells with trichostatin A, a specific HDAC inhibitor, attenuated Brg1-directed inhibition of reporter activity in transient transfection analysis. In addition, coimmunoprecipitation analysis showed that Brg1 interacted stably with the corepressor mSin3A and HDAC2 in erythroid cells, suggesting that HDAC activity is involved in Brg1-mediated transcriptional repression. Quantitative chromatin immunoprecipitation (ChIP) analysis demonstrated that Brg1 occupancy of the P4.2 and ß-globin promoters and hypersensitive site 2 (HS2) of the ß-globin LCR decreased upon erythroid differentiation. Importantly, the selective transcriptional repression of P4.2 and ß-globin gene expression by Brg1 correlated with increased loading of mSin3A and HDAC2 on the P4.2 promoter and ß-globin HS2, respectively. Together, these results suggest that Brg1-mediated transcriptional repression of select erythroid genes is characterized by the localized recruitment of a corepressor complex containing mSin3A and HDAC2 in a process of active repression.
Synergistic Interaction between Notch ligand Delta-1 and IL-6/Soluble IL-6 Receptor Fusion Protein in the Generation of BFU-E from Primitive Hematopoietic Progenitors.
Authors: Yamamura Kentaro , Ohishi Kohshi , Katayama Naoyuki , Shiku Hiroshi , Masuya Masahiro , Heike Yuji , Nishikawa Mitsuo , Nakahata Tatsutoshi 
Published: October 2019

Abstract
We have previously reported that Notch ligand Delta-1 may enhance the generation of NOD/SCID-repopulating hematopoietic stem cells and thymus-repopulating T cell precursors from cord blood [CB] CD34+CD38- cells. A complex of IL-6/soluble IL-6 receptor is shown to induce the expansion and proliferation of primitive hematopoietic progenitors that express gp130 but not the IL-6 receptor α chain, in mobilized peripheral blood (MPB) as well as in CB. In this study, we explored the interaction between Delta-1 and IL-6/soluble IL-6 receptor fusion protein [FP] in the generation of hematopoietic progenitors from MPB- and CB-derived primitive hematopoietic progenitors. MPB CD34+ Thy-1+ cells were cultured in serum-free medium with SCF, flt-3 ligand, TPO, and IL-3 [4GF], 4GF plus Delta-1, 4GF plus FP, and 4GF plus FP plus Delta-1. Delta-1 was used after immobilized to the culture plates. Freshly isolated cells and cultured cells were assessed for colony-forming ability by replating cells into semisolid medium containing SCF, flt-3 ligand, TPO, IL-3, IL-6, G-CSF, GM-CSF, and EPO. Colonies were counted at day 14. The numbers of BFU-E, CFU-Mix, and CFU-GM were 37-fold, 18-fold, and 135-fold increased, respectively, in cultures with 4GF plus FP, as compared with those in freshly isolated cells. The addition of both Delta-1 and FP to cultures resulted in 216-fold, 22-fold, and 132-fold increases in the numbers of BFU-E, CFU-Mix, and CFU-GM, respectively, relative to freshly isolated cells. Delta-1 did not affect the generation of colony-forming cells in cultures without FP. Thus, Delta-1 and FP synergistically enhanced the generation of BFU-E from primitive hematopoietic progenitors in MPB. In the cultures of CB CD34+CD38- cells with SCF, flt-3 ligand, TPO [3GF], 3GF plus Delta-1, 3GF plus FP, and 3GF plus FP plus Delta-1, the synergistic enhancement by Delta-1 and FP was observed in the generation of BFU-E and CFU-Mix. These data suggest a novel role for interaction between Notch and gp130 signalings in the generation of erythroid progenitors from primitive hematopoietic progenitors.
Low Dose Thalidomide and Donor Lymphocyte Infusion as Adoptive Immunotherapy after Allogeneic Stem Cell Transplantation in Patients with Multiple Myeloma.
Authors: Kroeger Nicolaus , Shimoni Avichai , Zagrivnaja Maria , Ayuk Francis , Lioznov Michael , Schieder Heike , Fehse Boris , Zabelina Tatjana , Nagler Arnon , Zander Axel 
Published: October 2019

Abstract
To improve anti-myeloma effect of donor lymphocyte infusion (DLI) after allogeneic stem cell transplantation in multiple myeloma, we investigated in a phase I/II study the effect of low dose thalidomide (100 mg) followed by DLI in 18 patients with progressive disease or residual disease and prior ineffective DLI after allografting. The median age was 53 years (range 31–64). Twelve patients received prior DLI and either relapsed again (n=1), or showed no response (PD: n=3; SD: n=8). The overall response rate was 67% including 22% complete remission. Major toxicity of thalidomide was weakness grade I/II (68%) and peripheral neuropathy grade I/II (28%). Only 2 patients experienced mild grade I acute graft versus host disease (aGvHD) of the skin, while no grade II-IV aGvHD was seen. De novo limited chronic GvHD (cGvHD) was seen in two patients (11%). The median time to response was 108 days (range, 36–266 days). In 5 patients not responding to 100 mg, the dose of thalidomide was increased (200 mg n=4; 300 mg n=1) and subsequently two of them responded with partial remission. In three patients dose escalation of DLI was performed, resulting in one minor and one partial remission. No difference regarding response was observed between unrelated and related donors (66% each). Five out of 6 patients with deletion 13 responded, while 5 out of 9 patients without deletion 13 responded to thalidomide plus DLI. In patients who failed to respond to prior DLI, the disease status could be converted in 6 patients with SD after first DLI into CR (n=2), PR (n=3), MR (n=1) while two with SD remainded SD after thalidomide plus DLI. In three patients with progressive disease after DLI the combination of thalidomide and DLI resulted in one CR, one PR and one minor remission.
The two-years estimated overall and progression-free survival was 100 % and 84%, respectively. Adoptive immunotherapy with low dose Thalidomide and DLI induces strong anti-myeloma effect with low incidence of graft versus host disease.
Thrombopoietin Regulates Proliferation and Maturation of Murine Mast Cells.
Authors: Migliaccio Giovanni , Ghinassi Barbara , Centurione Lucia , Zingariello Maria , Bianchi Lucia , Pancrazi Alessandro , Paoletti Francesco , Vannucchi Alessandro M. , Rana Rosa Alba , Nishikawa Mitsuo , Migliaccio Anna Rita 
Published: October 2019

Abstract
Megakaryocytopoiesis is regulated by extrinsic (interaction of the growth factor thrombopoietin, TPO with its receptor Mpl) and intrinsic (interaction between the trascription factors GATA-1 and Fog-1) factors. The observation that mice impaired for GATA-1 expression (i.e. harbouring the GATA-1low mutation) are defective not only in megakaryocyte maturation but also in mast cell differentiation (Migliaccio et al. J Exp Med197:281, 2003), led us to investigate whether TPO might control mast cell differentiation as well. We first observed that mice genetically unable to responde to TPO (Mplnull mice) express in the connective tissues 5 times more mast cells than their normal littermates. Then, we analysed the effects on mast cell differentiation of in vivo treatment with TPO. Normal mice, and their GATA-1low littermates, were injected i.p. with TPO (100 μg/kg/day per 5 days, kindly provided by Kirin Brewery, Japan) and the number of immature (Toluidinepos) and mature (AlcianBlue/Saphraninepos) mast cells present in the connective tissues of the animals, as well as the frequency of GATA-1pos and TUNELpos mast cells, was evaluated 14 days after treatment. In wild-type animals, TPO reduced the presence of GATA-1 in mast cells (by immuno-histochemistry) and increased the number of immature cells (from 320±28 to 852±60) and of those undergoing apoptosis (from 16±1 to 600±43). In contrast, in GATA-1low animals, TPO-treatment induced the expression of GATA-1 in mast cells while decreased the number of immature cells (from 1100±72 to 427±29) as well as that of apoptotic cells (from 600±45 to 60±2). The role of TPO on mast cell differentiation were further confirmed by the analysis of the effects exerted by the growth factor on in vitro differentiation of bone marrow derived mast cells (BMMC). In these experiments, wild type bone marrow and spleen cells were cultured for 21 days with SCF and IL-3 with or without TPO and BMMC differentiation measured on the basis of the number of cells expressing the phenotype c-kithigh/CD34high and FcεRIpos. In cultures stimulated with SCF and IL-3, all the cells expressed the phenotype c-kithigh/CD34high and FcεRIpos. In contrast, in cultures supplemented also with SCF, IL-3 and TPO, only 25% of the cells were c-kithigh/CD34high and none of them was FcεRIpos. These results establish a role for TPO in the control of mast cell differentiation (possibly by modulating the GATA-1 content of the cells) and unveil further similarities between the mechanism(s) controlling megakaryocyte and mast cell differentiation.
The Low-Molecular-Weight-Heparin, Tinzaparin, Is Effective and Safe in the Treatment and Prophylaxis of Venous Thromboembolic Disease during Pregnancy.
Authors: Jorgensen Maja , Nielsen Jorn D. 
Published: October 2019

Abstract
During pregnancy the incidence of venous thromboembolism (VTE) is approximately 6 times higher than in age-matched, non-pregnant women and venous thromboembolism remains the most common cause of maternal morbidity and mortality. Low-molecular-weight-heparins are recommended for treatment of VTE during pregnancy and for prophylaxis of VTE in pregnant women with major thromboembolic risk factors. We used tinzaparin for prophylaxis and treatment of VTE in 305 consecutive pregnant women referred to the Thrombosis Centre, Gentofte University Hospital, from 1997 to 2004. In 268 pregnancies the mothers had thrombophilia, 52 women were admitted with acute VTE and 184 had previous VTE. Other clinical risk factors included previous bad obstetric outcome, recurrent miscarriages, cardiac disorders or previous thromboembolic stroke. An individual risk assessment of each pregnant woman was performed. Very high risk females were treated with tinzaparin 90 – 100 IU/kg bid, high risk females were treated with tinzaparin 100 – 125 IU/kg daily and women with moderate risk were treated with 50 – 75 IU/kg daily.
302 of 305 pregnancies (99 %) in 263 females resulted in 310 healthy babies. 306 of 310 babies had appropriate birth weight for gestational week and all babies had normal Apgar score. Two females had miscarriages (week 10 and 20) and 1 female had an elective abortion. No females had pulmonary embolism. Deep venous thrombosis occurred in 4 of 305 pregnancies = 1,3 % (week 6, 11, 27 and one day postpartum). Wound hematoma was observed after cesarean section in two women and postpartum bleeding episodes (700 – 1500 ml) were observed in 7 women (4 had severe vaginal or cervical tearings, 2 had retained placenta and 1 had placental abruption). We found no incidence of thrombocytopenia or symptomatic osteoporosis.
We find that individually dosed tinzaparin is safe and seems effective in the prevention of thromboembolic complications during pregnancy. Individual risk stratification is recommended.
ZAP-70 Expression in Chronic Lymphocytic Leukemia: Correlation with Clinical and Biological Features.
Authors: Del Giudice Ilaria , Osuji Nnenna , Matutes Estella , Morilla Alison , Morilla Ricardo , Burford Anna , Maravelaki Sonia , Owusu-Ankomah Kwasi , Swansbury John , Brito-Babapulle Vasantha , Catovsky Daniel 
Published: October 2019

Abstract
One of the most intriguing features of chronic lymphocytic leukemia (CLL) is its clinical heterogeneity. The mutational status of immunoglobulin heavy chain variable region (IgVH) genes is one of the most powerful predictors of overall survival (OS) and progression-free survival (PFS). The expression of the tyrosine kinase ZAP-70 in CLL cells has been proposed as a surrogate marker for the mutational status of IgVH genes. Recent reports suggest that ZAP-70 over-expression is an independent prognostic factor for shorter OS and PFS. There is little information about correlation between ZAP-70 expression and other clinical and biological features in CLL. The aims of this study are 1) to evaluate prospectively ZAP-70 expression in a series of CLL patients; 2) to correlate this with clinical stage, lymphocyte morphology, CD38 expression and chromosomal abnormalities detected by FISH and 3) to analyze the independent prognostic value of ZAP-70 in predicting time to first treatment (treatment free interval, TFI). ZAP-70 expression was analyzed using four-colour flow cytometry (as described by Crespo et al, NEJM2003; 348:1764–75). 201 previously untreated CLL patients were evaluated at diagnosis (32%) or during the disease course. Amongst them, 143 patients were entered in the LRF CLL4 trial. Clinical stage at the time of the study was A in 92 patients (A progressive in 46), B in 63 and C in 46. 28% were ZAP-70 positive (+) (cut-off ≥ 20% of CD5/CD19+ cells). ZAP-70 positivity was associated with prevalence of stage B/C at diagnosis (p=0.004), CLL with >10% prolymphocytes (CLL/PL) or atypical morphology (p<0.001), CD38 positivity (regardless of the cut-off point) (p<0.001), trisomy12 (p=0.001), del(6)(q21) (p=0.05) or no detectable abnormalities (0.002). ZAP-70 negative (−) cases presented more often with del(13)(q14) (p<0.001). There were no significant differences in the frequency of del(11)(q23) or delp53 between ZAP-70(+) and ZAP-70(−) cases. Median time from diagnosis to first treatment (treatment free interval, TFI) was 17.7 months for ZAP-70(+) and 44.6 months for ZAP-70(−) cases (p<0.001). The presence of the following parameters was associated with short TFI in univariate analysis: male sex (p=0.02), atypical morphology (p=0.02), clinical stage B/C (p<0.001), ZAP-70(+) (p<0.001), CD38+ (p<0.001), absence of del(13)(q14) (p<0.001), presence of delp53 (p=0.05). Multivariate analysis is being performed. In conclusion, we document a significant correlation between ZAP-70 over-expression in CLL and other adverse prognostic factors (such as CD38 and advanced stage), provide new data concerning its association with cytogenetic abnormalities detected by FISH and demonstrate the prognostic value of ZAP-70 expression.
Embryonic Stem Cell Transplant (ESCT)-Derived Mixed Hematopoietic Chimerism Prevents Diabetes in NOD Mice.
Authors: Burt Richard K. , Verda Larissa , Kim Duck A. , Statkute Laisvyde , Oyama Yu , Krosnjar Nela , Ikehara Susumu 
Published: October 2019

Abstract
NOD mice are a widely used animal model for studying type I diabetes mellitus characterized by lymphocytic infiltration of pancreatic islets following by development of diabetes by age 3 to 4 months. It has been shown that allogeneic bone marrow transplantation can prevent insulitis and overt diabetes. We have recently demonstrated that embryonic stem cell-derived hematopoietic stem cells (ESC-derived HSC) can reconstitute bone marrow in lethally irradiated mice across MHC barriers without graft versus host disease (GVHD). Herein we report a new ESC-derived HSCT approach for the prevention of autoimmune diabetes in NOD mice. 6-weeks old female mice were sublethally irradiated (8.0 Gy) and transplanted with ESC-derived HSC. To induce differentiation toward HSC in vitro, ESC (R1) were cultured in methylcellulose-based medium supplemented with SCF, IL-3 and IL-6. An enriched c-kit+CD45+ ESC-derived suspension was delivered intra bone marrow (IBM) (mean 0.5 x 106/mouse) (N=8) or intravenously (mean 1 x 106/mouse) (N=6). Six NOD mice were held as control. Mice were followed by blood glucose measurements and chimerism analysis until onset of diabetes or until 32 weeks after transplantation. 7 out of 8 mice from IBM group and 4 out of 6 from IV group did not become hyperglycemic in contrast to control group where 5 out of 6 mice were euthanized because of diabetes. The level of chimerism achieved after transplantation was 8.2%±5.76 in IBM group and 3.2%±2.75 in IV group. Histological examination showed that most of islets were replaced by lymphocytic infiltration or fibrous tissue in controls (even in case of a mouse without clinical evidence of diabetes). In 78% (11/14) of animals from ESC-derived hematopoiesis, remission was confirmed by histology revealing absence of insulinitis and normal immunohistochemical staining of islet cells for insulin. Prevention of diabetes/insulinitis was predicted by the percentage ESC-derived hematopoietic chimerism. All mice with > 5% ESC-derived chimerism remained free of diabetes and insulinitis. These results suggest that allogeneic chimerism achieved with embryonic stem cells can prevent diabetes in NOD mice without evidence of GVHD or use of post transplant immune suppression.