bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cooperative Silencing of Chicken ρ- and βH- Globin Gene Expression by Chicken β-Globin Regulatory Elements: Involvement of RNAi Mediated Repression.
Authors: Epner Elliot M. , Wang Jin , Huang Jing 
Published: October 2019

Abstract
The chicken β-globin locus represents a well characterized, model system where the relationship between chromatin structure, transcription and DNA replication can be studied. The locus contains several regulatory elements including an intergenic enhancer as well as upstream regulatory elements that may function either alone or in combination with the intergenic enhancer as an LCR. The availability of the recombination proficient chicken B cell line DT40 has allowed the introduction of mutations into the endogenous chicken β-globin locus and phenotypic analysis after microcell mediated chromosome transfer into human erythroleukemia (K562) cells. Using this system, we have introduced deletions in the chicken β-globin intergenic enhancer as well as 5′ HS 1,2, and 3. Expression of the embryonic ρ and fetal βH chicken globin genes were repressed by the intergenic enhancer, 5′ HS1, or 5′HS2. No ρ or βH globin gene expression was detected in K562 cells containing control chicken chromosomes, while ρ and βH mRNA were activated when the intergenic enhancer, 5′ HS1, or 5′HS2 were deleted. Chromatin immunoprecipitation (ChIP) experiments that assayed RNA polmerase II (pol II), GATA-1 and NF-E2 p45/ p18 binding at regulatory elements and gene promoters in targeted cell lines supported this hypothesis and suggested a potential role for 5′HS3 in gene activation. However, targeted deletion of 5′ HS3, unlike the other chicken β-globin regulatory elements, showed no transcriptional phenotype. Our results demonstrate the intergenic enhancer, 5′HS1, and 5′ HS2 function through a common silencing mechanism involving pol II, GATA-1, and NF-E2/P18. The recent demonstration of the involvement of Pol II in the synthesis of miRNA’s prompted us to investigate the role of miRNA’s in gene silencing in this system. A small miRNA was identified at the intergenic enhancer region. ChIP assays showed the binding of two components of the RISC (Dicer and Ago2) at the chicken globin regulatory elements. These results are consistent with the involvement of RISC and miRNA’s in gene silencing in this system.
Over-Expression of Prolylcarboxypeptidase Enhances Plasma Prekallikrein Activation on Chinese Hamster Ovary Cell Membranes.
Authors: Shariat-Madar Zia , Rahimy Ehsan , Mahdi Fakhri , Schmaier Alvin H. 
Published: October 2019

Abstract
Recent investigations indicate that there is a physiologic plasma prekallikrein (PK) activator on the membrane of endothelial cells (HUVEC). PK complexes with high molecular weight kininogen (HK) and HK is PK’s receptor on HUVEC. When assembled on HUVEC, PK is activated to plasma kallikrein independent of FXIIa by the serine protease prolylcarboxypeptidase (PRCP) (Km=9 nM). PRCP has been shown to be a PK activator when isolated from HUVEC (JBC277:17962, 2002) and produced as a recombinant protein (Blood103:4554, 2004). To additionally confirm that human PRCP is a physiologic PK activator, PRCP was over-expressed in Chinese hamster ovary cells (CHO). CHO were transfected with full-length PRCP under the control of a CMV promoter and CHO rPRCP was expressed as a fusion protein with C-terminal enhanced green fluorescent protein (EGFP). The presence of rPRCP in transfected CHO was detected by real time RT-PCR, immunoblot and immunoprecipitation. In CHO cells, PRCP mRNA and PK activation are 2–3-fold higher than controls. The increase in PRCP-induced PK activation on the transfected CHO cells parallels the increase in PRCP antigen expression on the transfected cells, as determined by anti-PRCP and anti-GFP antibodies. PK activation of the transfected cells is blocked by siRNA to the translation initiation site of PRCP, anti-PRCP antibody, and Z-Pro-Pro-aldehyde dimethyl acetate, with IC50 of the last 2 inhibitors of 0.01 and 7.0 mM respectively. Cellular localization of PRCP in intact cells using confocal microscopy and flow cytometry indicate that its over-expression results in its placement on the CHO cell plasma membrane. These investigations independently confirm that prolylcarboxypeptidase is expressed on cell membranes and its expression increases PK activation. The endothelial cell membrane expression of PRCP resulting in PK activation contributes to the constitutive level of bradykinin in the intravascular compartment to regulate blood pressure homeostasis.
Low Levels of Zinc and Anti-Oxidant Vitamines (A, C & E) in Patients with Sickle Cell Anemia.
Authors: Hasanato Rana M. 
Published: October 2019

Abstract
Background : Patients with sickle cell anemia have a higher potential for oxidative damage due to chronic redox imbalance in their red blood cells that often leads to hemolysis, endothelial injury and recurrent vaso-occlusive episodes. In this study, we evaluated plasma levels of the anti-oxidant vitamins A, C,& E and serum level of Zinc along with serum level of Copper (pro-oxidant ).
Patients and methods: 25 adult patients with documented severe sickle cell anemia and frequent painful episodes (12 males & 13 females, aged 29.27 +/− 12.94 years) and 25 matched normal controls were studied. Plasma levels of Vitamins A, C, & E were measured by HPLC technique and serum levels of zinc & copper were measured by atomic absorption spectrometry.
Results: (Table ): Conclusion These results indicate that levels of Vitamines A, C, and E and Zinc are significantly low in patients with sickle cell anemia in comparison with matched normal controls. Besides, Copper level, which is a well known pro-oxidant, is significantly higher in sickle cell anemia patients. Therefore, supplementing sickle cell anemia patients with anti-oxidant vitamins and trace elements may contribute to the amilioration of symptoms and severity of sickle cell anemia
Levels of Vitamine A, C, & E , Zinc & Copper in patients with Sickle cell anemia and normal controls Tested Items SCS patients Control group P-value Results are shown as mean +/− standard deviation., SCA = Sickle cell anemia Vitamin A level mcg/l 0.13 +/− 0.03 0.33 +/−0.05 <0.0001 Vitamin C level mg/l 9.00 +/− 3.43 20.32 +/−3.74 <0.0001 Vitamin E level, mcg/l 1.95 +/− .0.69 4.30 +/−0.80 <0.0001 Zinc level mmol/l 9.00 +/− 1.89 11.81 +/−1.93 <0.0001 Copper level mmol/l 24.58 +/− 4.87 18.42 +/−3.06 <0.0001
Expression of Transcription Factors NFAT1, c-Jun, c-Fos, and Bach2 in Umbilical Cord Blood and Adult Blood T-Cells.
Authors: Huang Ying , Leahy Patrick , Kaminski Beth A. , Daum-Woods Kathy , Veigl Martina , Laughlin Mary J. 
Published: October 2019

Abstract
Background: Nuclear factor of activated T cells 1 (NFAT1 or NFATc2) is proposed to plays a role in Graft versus host disease (GVHD) because it regulates the transcription of many cytokines and surface regulatory proteins including: cytotoxic T lymphocyte-associated antigen 4 (CTLA-4), IL-13, IFN-γ, CD40L, granulocyte-macrophage colony-stimulating factor (GM-CSF), and TNF-α. NFAT1 regulates the expression of these genes through direct interaction with the leucine zipper region of AP-1 (Fos/Jun) and through cooperative binding to a 15-bp DNA sequence that contains both NFAT1 and AP-1 sites. The highly conserved basic regions of AP-1 (Fos/Jun heterodimer) bind to the TGTTTCA consensus sequence in DNA. Human Bach2 is also a basic-leucine zipper (bZip) protein. The amino acid residues in contact with the DNA in the basic regions of c-Jun and c-Fos are also identical in Bach2 (human and mouse). The high conservation of DNA-contacting amino acid residues between Bach2 and AP-1 strongly suggests that Bach2 (homodimers) and AP-1 (Fos/Jun heterodimers) can bind to the same DNA site, which may block the formation of NFAT1/AP-1/DNA complex and the subsequent expression of cytokine genes. Clinical studies have determined that umbilical cord blood (UCB) elicits reduced incidence and severity of GVHD, compared to Bone Marrow (BM), due in part to reduced donor T-cell cytokine production.
Methods: We compared the RNA and protein levels of NFAT1, c-Jun, c-Fos, and Bach2 in human UCB CD4+ T-cells with Adult Blood (AB) CD4+ T-cells by Quantitative RT-PCR analyses and Western blots. This comparison was done in both stimulated and non-stimulated conditions. Our ongoing interest is to test whether Bach2 binds the same DNA site as AP-1, and to test the potential of Bach2 and AP-1 competing for the same DNA site using Electrophoretic mobility shift assay (EMSA) with purified Protein of Bach2, c-Jun, and c-Fos from E. coli.
Results: Our data showed a high expression of Bach2 RNA in both non-stimulated and stimulated UCB CD4+T-cells. In both the non-stimulated comparison and stimulated comparison samples, we detected no significant changes in RNA levels of NFAT1, c-Jun, or c-Fos.
Conclusions: Our data support the hypothesis that Bach2 acts as a transcriptional repressor of cytokine genes due to the fact that highly expressed Bach2 in UCB competes with AP-1 for same DNA binding site.
Clinical Application Study of Plasma Thrombmodulin.
Authors: Lu Xingguo , Tong Jiefeng , Zhang Qiang , Zhang Mao , Zhao Xiaoying , Fu Gousheng , Xu Genbo 
Published: October 2019

Abstract
Objective: To investigate the clinical value of plasma thrombmoclulin (PTM).
Methold: PTM was determined in 979 patients and 60 healthy controls with ELISA method.
Result: The PTM of the control group was 20.40±7.72 μg/L, there was no difference in genders and ages. The PTM of the patients with microangiopathy were higher than that of the patients with macroangiopathy (P <0.01). We defined PTM level higher than its normal upper limit (35.54μg/L) as PTM positive, in microangiopathy diseases, the sensitivity of PTM was 77.7%, the specificity was 71.2% and the positive predictive value was 75.6%. In chronic primary glomerular disease, the PTM of chronic renal failure (CRF) group was higher than that of non-CRF group (P <0.01), the sensitivity was 85.7%, the specificity was 82.4%, the positive predictive value was 77.8%. The PTM level in septemia group was higher than that of non-septemia group(P <0.01),the sensitivity of PTM was 86.6%, the specificity was 89.5% and the positive predictive value was 76.5%. In multiple trauma, the PTM of multiple organ failare (MOF) group was higher than that of non-MOF group (P <0.01), the sensitivity of PTM was 77.8%, the specificity was 77.3% and the positive predictive value was 73.7%. In systemic lupus erythematosus (SLE), the PTM of the patients with albuminuria was higher than that of the patients without albuminuria(P <0.01), the sensitivity was 77.8%, the specificity 92.3% and the positive predictive value was 93.3%. In diabetes, the PTM of complication group was higher than that of no complication group, the sensitivity was 53.4%, the specificity was 97.1% and the positive predictive value was 98.6%, the PTM of microangiopathy group was higher than that of macroangiopathy group (P <0.01), the sensitivity was 71.2%, the specificity was 97.1%, and the positive predictive value was 97.9%. Compared the acute stage with restoration stage in stroke, pre-chemotherapeutics with post-operation in AL and MM, and pre-operation with post-operation in cancer, the PTM was connected with clinical development.
Conclusion: PTM was a good criterion in evaluating the microangiopathy in clinical application. PTM was also a valuable index in prediction or assessment of the severity of diseases or in the observation of therapeutic effectiveness.
Thoracal Carcinoma Surgery in FVII Deficient Patient Managed with rFVIIa.
Authors: Erdlenbruch Kerstin , Stephan Bernhard , Bakchoul Tamam , Schenk Joachim , Pindur Gerhard 
Published: October 2019

Abstract
Introduction: In patients with congenital FVII deficiency and positive bleeding history surgical procedures should be covered by an sufficient substitution therapy of FVII concentrate. For the first three postoperative days trough level of at least >0,2U are recommended. We report haemostasis management of a complex thoracic surgery procedure in a 14 year old boy with FVII deficiency and diagnosis of a mucoepidermoid bronchogenic carcinoma of the left upper lobe. Extended thoracic surgery without extra corporal circulation was performed as a lobectomy of the left upper lobe including extended preparation of lymphnodes.
Preoperative Lab:
Platelets 285000/μl, INR 1,6, aPTT 28s, fibrinogen 239 mg/dl, FVII 0.33IU
Because of positve bleeding anamnesis, the extended surgical procedure and possibility of further extension of the planned procedure we decide to cover surgery by factor treatment to be on the safe side though FVII activity was 0.33IU. Because of virus safety and safety profile we decide to give rFVIIa (NovoSevenR) as bolus injections. Frequency of treatment was chosen according to regular lab controls and clinical course.
Haemostasis management during surgery:
Preoperative rFVIIa dosage: 30 μg/kg BW, given immediately before skin cut as a bolus. Intraoperativly was no significant bleeding and no transfusion requirements though it was a complex and extended procedure.
Haemostasis management postoperatively:
Regular control of FVII activity and Hb was done every 6 hours to decide if substitution therapy is required. In the early phase after surgery FVII activity remained >1.0 IU, so additional factor treatment was not necessary. Hb remained stable in normal range. 24h after surgery FVII activity fell to 0.16IU. Substitution was done with dosage of 15μg/kgBW rFVIIa as a bolus (NovoSevenR). Frequency: once daily, duration: 2 days Clinical course: no bleeding complications. For remove of drainges a last bolus of 30μg/kg was given. Residual FVII activity: >0,2IU in all regular controls during the first three days.
Figure 1: thromboplastin time, F VII activity and tFVIIa substitution in the course of time Figure 1:. thromboplastin time, F VII activity and tFVIIa substitution in the course of time
Results:
Bleeding/Transfusion requirements: No bleeding complications or wound healing complications occurred, no blood products has been transfused.
Adverse events: no adverse events, no thrombembolic events and no inhibitor development.
Conclusions: Regular control of FVII activity and clinical bleeding signs led to an efficient use of factor concentrates. rFVIIa given as bolus in FVII deficiency seem to be a save and efficient therapy for haemostasis management in complex thoracic cancer surgery.
Self-Managed Long-Term Low-Molecular-Weight Heparin Therapy Compared with Usual-Care Anticoagulation for the Treatment of Patients with Proximal Venous Thrombosis: Harm Associated with Thrombocytopenia.
Authors: Hull Russell D. , Pineo Graham F. , Brant Rollin F. ,   for the LITE Trial Investigators
Published: October 2019

Abstract
Context: A substantial clinical need exists for an alternate to vitamin-K-antagonists for treating deep-vein thrombosis in many patients. Long-term low-molecular-weight heparin, body-weight adjusted, avoids anticoagulant monitoring and may be associated with less harm due to thrombocytopenia.
Objective: Considerable evidence suggests that reporting of harms-related data* from randomized clinical trials needs improvement. In accordance with CONSORT* we present a harm analysis regarding thrombocytopenia.
Design: Randomized clinical trial using objective outcome measures.
Setting: 30 centres across Canada.
Participants: Acute symptomatic proximal-vein thrombosis patients.
Intervention: Therapeutic tinzaparin subcutaneously once-daily, or intravenous unfractionated heparin/oral vitamin-K-antagonist therapy for 3 months.
Main Outcome Measures: Benefit was assessed by assessing the frequency of symptomatic objectively documented recurrent venous thromboembolism and harm by objectively documented hemorrhagic complications and the outcome of thrombocytopenia.
Results: Benefit and bleeding harm are reported elsewhere. Long-term low-molecular-weight heparin treatment was at least as effective but associated with less harm due to bleeding than usual care. Thrombocytopenia occurred in 21 of 369 (5.7%) patients receiving tinzaparin versus 9 of 368 (2.4%) patients receiving usual care (absolute difference 3.3). Six of 9 patients with thrombocytopenia receiving usual care died (66.7 percent, 95 percent confidence interval 29.9 to 92.5) versus 4 of 21 with thrombocytopenia receiving low-molecular-weight heparin (19 percent, 95 percent confidence interval 5.4 to 41.9 percent) (absolute difference 47.6 percent, 95 percent confidence interval 82.7 to 12.5 percent, p=0.03). Recurrent venous thromboembolism occurred infrequently in patients with thrombocytopenia.
Conclusion: Our findings suggest that harm associated with thrombocytopenia in patients receiving intravenous unfractionated heparin versus low-molecular-weight heparin favors low-molecular-weight heparin use. Further evaluation in rigorous trials is required.
TGF-β1 Affects the Proliferation of Normal Human Bone Marrow Fibroblasts and Its Expresssion of Smad3 and Smad7 mRNA.
Authors: Huang Jinwen , Jin Xing 
Published: October 2019

Abstract
Current drug therapy in myelofibrosis with myeloid metaplasia (MMM) still can not get great progress, the development of the myelofibrosis eventually causes bone marrow failure. The histopathology of MMM patients showed that the bone marrow tissue were full of marrow fibroblasts and an extensive deposition of collagens. Further researches suggested that the proliferation of the fibroblasts was controlled by many factors. One of the most important factor was TGF-β1, and its effects largely depended on Smad pathway. In order to understand the effects of TGF-β1 on the proliferation of marrow fibroblasts, we stimulated the normal human bone marrow fibroblasts with different concentrations of TGF-β1 in the current study. The proliferation test with direct cell counting and MTT assay revealed that different concentrations of TGF-β1 had various effects and it could get the maximal proliferation effect at 0.1ng/ml. Then we detected the Smad3 mRNA and Smad7 mRNA in one normal human bone marrow fibroblasts with RT-PCT methods at above concentration (0.1ng/ml). We found that the expression level of Smad3 mRNA decreased with the time, and that the expression level of Smad7 mRNA reached the highest at 30 min after stimulation by TGF-β1, then it began to reduce. These results suggested that TGF-β1 activate the Smads pathway in normal human marrow fibroblasts, and its proliferation effect require the activation of this pathway. We also noticed that the changes of the smad3 mRNA and smad7 mRNA were similar in skin fibroblasts stimulated by TGF-β1. These changes were considered as a feedback modulation mechanism. Whether the over-proliferation of marrow fibroblasts in MMM patients is correlated with the imbalance of this feedback warrants further research. It may provide a more specific and targeted therapy for blocking myelofibrosis.
Fig A The expression of Smad3 mRNA stimulated by TGF-β 1(0.1ng/ml) at different time. Fig B The expression of Smad7 mRNA stimulated by TGF-β 1(0.1ng/ml) at different time. Fig A The expression of Smad3 mRNA stimulated by TGF-β 1(0.1ng/ml) at different time. . / Fig B The expression of Smad7 mRNA stimulated by TGF-β 1(0.1ng/ml) at different time.
Molecular Responders (<3-log Reduction) among CML Patients in Complete Cytogenetic Remission Show a Lower Number of BCR-ABL+ Hematopoietic Progenitors Compared to Non-Responders.
Authors: Ibatici Adalberto , Fugazza Giusy , Sessarego Nadia , Podesta Marina , Agliano Stefania , Pozzi Sarah , Corselli Mirko , Garuti Anna , Sessarego Mario , Frassoni Francesco 
Published: October 2019

Abstract
Background. BCR-ABL+ leukaemic hematopoiesis in CML patients treated with the tyrosyn kinase inhibitor Imatinib mesylate can remarkably diminish up to undetectable levels by cytogenetic or molecular analysis. This treatment seems to target leukemic progenitors cells while sparing some quiescent BCR-ABL+ leukaemic stem cells. In this study, we investigated the number of residual BCR-ABL+ hematopoietic progenitors in bone marrow (BM) samples from CML patients who reached a complete cytogenetic remission (CCR) with Imatinib.
Materials and Methods. BM samples (15–20ml) were obtained from 18 CML patients in CCR under Imatinib treatment. All of them were in 1st chronic phase when Imatinib was started. A CD34+ cell-enriched population was selected from BM mononuclear cells using a double immunomagnetic column separation. CFCs and LTC-ICs were obtained by seeding the CD34+ selected population using standard culture methods. FISH studies for the BCR-ABL fusion gene (Dual color, Dual Fusion DNA probe form Vysis, INC, Downer Grove, IL) were performed on freshly selected CD34+ cells, pooled CFCs and LTC-ICs. A total of 100–200 nuclei were scored for each sample. The normal cut-off value used for these probes (0–2%) was established by our lab validation studies. MMR were defined as BCR-ABL/ABL ratios less than 0.05% (roughly equivalent to a 3-log reduction). Real-time QRT-PCR was performed using a TAQ-Man system (ABI Prism 7700 Perkin Elmer, CA) for BCR-ABL and ABL genes. Results. The median number and purity of selected CD34+ cells was 0.51 (0.17–2.1) x 106 cells and 95% (72–99). MMR was present in 10/18 (55.5%) CML patients. The median follow-up for patients in MMR or not in MMR was 34 (14–54) and 36 (6–59) months, respectively. None of the MMR-CML showed residual BCR-ABL+ cells detected by FISH on freshly selected CD34+ cells (MMR vs not MMR P=.0039 by Fisher’s test), CFCs and LTC-ICs. Conversely, 6 out of the 8 CMLs not in MMR showed residual BCR-ABL+ CD34+ cells, CFCs and LTC-ICs.
Conclusions. According to previous studies (Blood2003; 101: 4701–7), we found that CML patients in CCR under Imatinib may have residual BCR-ABL+ hematopoietic progenitors. However, the size of leukemic progenitors may differ substantially between MMR and not MMR. CML patients with more than 3-log reduction of disease show undetectable BCR-ABL+ progenitors (CD34, CFC and LTC-IC) by FISH analysis. QRT-PCR on bone marrow samples may provide a reliable estimation of the number of residual leukaemic progenitors and may identify different clusters of CML patients in CCR.