bluuebunny/crossref_metadata_2025_split huggingface dataset:
Alterative Expression Pattern of T Cell Immunosuppressive Receptors in Peripheral Blood of Patients with ITP
Authors: Zhong Jun , Chen Shuaohua , Lai Jing , Xu Ling , Zhang Tao , Yu Zhi , Lu Yuhong , Yang Lijian , Wu Xiuli , Li Bo , Li Yangqiu 
Published: October 2019

Abstract
Immune Thrombocytopenia (ITP) is an autoimmune disorder characterized by production of auto-antibodies against platelet antigens. Dysregulation of T cells is a major role involved in this disease, while anti-platelet antibodies induce platelets destruction due to an imbalanced immune response. Increasing data showed that abnormal expression of T cell immunosuppressive receptors such as Programmed death 1 (PD-1) and Cytotoxic T lymphocyte antigen-4 (CTLA-4) may relate to autoimmune disease pathogenesis. In this study, we analyzed the expression levels of T cell immunosuppressive receptors including PD-1, PD-L1, CTLA-4, T cell immunoglobulin mucin 3 (Tim-3), lymphocyte activation gene-3 (LAG-3) and B and T lymphocyte attenuator (BTLA) in RNA from peripheral blood mononuclear cells (PBMCs) of ITP patients (18 cases, male: 12, female: 6, median age: 38.5 years, range 24-68 years), PBMCs from 20 healthy individuals were served as control group, by real-time PCR. The results showed that significant lower expression levels of PD-1 (median: 0.0015) and PD-L1 (median: 0.0572) were showed in ITP group in comparison with healthy group (PD-1, median: 0.0117, p < 0.0001; PD-L1, median: 0.5428, p < 0.0001), while the expression levels of Tim-3 (median: 0.1352), LAG-3 (median: 0.1403) and BTLA (median: 0.1403) were significant higher than that from healthy group (Tim-3, median: 0.0751, p < 0.007; Lag-3, median: 0.0155, p < 0.0001; BTLA, median: 0.0315, p < 0.0001). There was not significant different on the CTLA-4 expression level between ITP group (median: 0.0818) and healthy group (median: 0.1667) (p = 0.219). In conclusion, we firstly described the alterative expression pattern of T cell immunosuppressive receptors in ITP, which may have a role in ITP pathogenesis.

Disclosures
Li: The National Natural Science Foundation of China (81370605): Research Funding.
Bone Marrow Findings in Veterans (V) with Hepatitis C Infection (HCV) at a VA Medical Center
Authors: Srinivas Shanthi , Kim Katherine , Gonzalez Melanie , Zhong Fengming , Crump Barbara , Victor Chang 
Published: October 2019

Abstract
Background: Hepatitis C Virus (HCV) is a major public health problem, with an increased prevalence in Veterans. HCV has been associated with B cell Lymphoproliferative disorders and Type II and III Cryoglobulinemias.Prior studies also found that patients with HCV are more likely to develop cytopenias compared normal. Little is known about other possible hematologic manifestations of HCV.We present a series of 54 Veterans seen in a hematology oncology clinic with HCV and their hematologic and bone marrow findings
Methods: In an IRB-approved protocol, we reviewed the records of V with HCV who underwent bone marrow biopsy from January 1993 to July 2015. V with hematological malignancies, coinfection with human immunodeficiency virus (HIV), hepatitis A or B, were excluded. Demographics, Vietnam era status, clinical and pathological data, indications for bone marrow biopsies, and results were abstracted. Statistical analysis was performed using SAS 9.4.
Results: Of the 54 V, 30 Caucasians, 22 African Americans and 2 Hispanics met the eligibility criteria. 37 served in the Vietnam era. The median (M) age was 59 years (range 39-90). M for WBC 6.9 x109/L (2.9-36.3), hemoglobin 14.3 gm/dL (6.9-19), MCV 93.1 fL (68.8-110..6), platelets 172 x109/L (39-493) , total bilirubin 0.75 mg/dl (0.1-5.3), alkaline phosphatase 96.5 U/L (56-341), albumin 3.9 g/dl (1.6-4.9), lactate dehydrogenase 191 IU/L (91-867), AST U/L (19-426), ALT 56.5.U/L (4-480), INR 1.09 (0.84-2.04). 12 V had splenomegaly by imaging. 10 V at the time of bone marrow biopsy had cirrhosis. The most common indication for bone marrow biopsy was cytopenia (n=38) in at least one of the lineages. The remaining (n=16) were for paraproteinemia, lytic lesion, leukocytosis or lymphocytosis. 48 V had abnormal bone marrow biopsy findings (summarized in Table 1) and 6 V had normal biopsy findings.Table 1.Bone Marrow Findings (n=48)Dysplasia15Hyperplasia14Benign Lymphoid Aggregate4Plasma Cells3Leukemia, Plasma cell dyscrasia4Combination of hyperplasia, lymphoid aggregates or plasma cells8
Conclusions: Veterans with HCV develop cytopenias and hematological malignancies. While Cytopenias can result from splenomegaly, autoimmune mechanism, and medications, some of the cytopenias are unexplained Bone marrow changes from chronic HCV infection may be a contributing factor. Understanding these bone marrow changes from HCV may help in determining the etiology and further management of hematological problems related to HCV

Disclosures
No relevant conflicts of interest to declare.
The Comparison of Hemostatic Effects of Recombinant Activated Factor VII (rFVIIa) in Thrombocytopenic Patients (pts) and Hemophilia Pts with Inhibitors
Authors: Galstian Gennadii M. , Kolosova Irina V. , Gemdzhian Eduard , Zorenko Vladimir Yu. , Polyanskaya Tatyana A. , Orel Elena B. , Savchenko Valeri G. 
Published: October 2019

Abstract
Background: Administration of rFVIIa is used as a method of choice in the treatment of hemophilia pts with inhibitors. Besides, rFVIIa may be useful in thrombocytopenic pts with inefficiency or impossibility of platelet transfusions due to alloimmunization or religious beliefs.
Aim: The aim of the present study was to compare the effects of rFVIIa on hemostasis in thrombocytopenic pts and hemophilia pts with inhibitors.
Patients and Methods: rFVIIa was used in 20 pts with thrombocytopenia (I group) and in 20 hemophilia pts with inhibitors (II group). Indications for use of rFVIIa in thrombocytopenic pts were: gastrointestinal and intra-abdominal bleeding, intracranial hemorrhage, insertion of the central venous catheter, lumbar puncture, bleeding from the femoral artery puncture site, epistaxix and pulmonary hemorrhage. In thrombocytopenic pts blood platelet count varied from 1*109/l to 72*109/l (median 15 *109/l). The media dose of rFVIIa in thrombocytopenic pts was 79 μg/kg (ranges from 50 to 144 μg/kg). All hemophilia pts received a single dose of rFVIIa (90 μg/kg). All pts received rFVIIa (Generium, Russia). APTT, Quick prothrombin test (QPT), FVII:C level, endogen thrombin potential (ETP) and Thromboelastography (TEG) parameters (R, MA) were evaluated before and after rFVIIa administration.
The median and interquartile range (IQR) were calculated as descriptive statistics. Mann-Whitney U test and Mood's median test were used to evaluate group differences. Statistical analyses were performed with SAS 9.1 (using the Npar1way procedure).
Results: rFVIIa was effective in all hemophilia pts. Administration of rFVIIa in 15 min shortened but did not correct APTT, increased QPT and ETP, FVII:C (Table 1). Prior to treatment the hemophilia pts have no clotting determined by TEG, but after rFVIIa administration TEG demonstrated close to normal pattern. In thrombocytopenic pts after rFVIIa administration bleeding stopped in 51% pts, decreased in 44% and did not change in 4% pts. In thrombocytopenic pts maximal hemostatic effect was achieved within 1 hour after rFVIIa administration (Table 1). 1 case of ischemic stroke was obtained. The maximum effect of rFVIIa in thrombocytopenic pts coincided with decrease of plasma activity of FVII perhaps due to its consumption.
Conclusion: In the most hemophilia pts with inhibitors the response was achieved within 15 min of administration of rFVIIa. In thrombocytopenic pts haemostatic effect of rFVIIa was delayed and reached within 1 hr, good response was achieved in 51% pts.
Figure 1. Parameters of hemostasis before and after rFVIIa administration expressed as a median (IQR) Figure 1. Parameters of hemostasis before and after rFVIIa administration expressed as a median (IQR)

Disclosures
No relevant conflicts of interest to declare.
Identification of Molecular Defects in ITGA2B and ITGB3 Genes and Phenotypic Correlation in Pakistani Patients with Glanzmann Thrombasthenia
Authors: Jamal Yonus , Ahmed Shariq , Najmuddin Akbar , Imran Ayisha , Khan Tehmina nafees sonia , Shamsi Tahir , Naz Arshi 
Published: October 2019

Abstract
Background:
Glanzmann thrombasthenia (GT) is most common inherited platelet functional defect. It is an autosomal recessive disorder, characterized by a bleeding diathesis. Incidence is increased in locations where consanguineous marriages are common. The defect is caused by mutations in the genes encoding ITGA2B or ITGB3. This results in qualitative or quantitative abnormalities of the platelet receptor, αIIb-β3 integrin.
Objectives:
The aim of this study was to identify and correlate the mutations in GT patients with phenotype of the patient.
Subjects and methods:
20 patients with GT were enrolled in the study to identify the molecular defects and to correlate their phenotype with their genotype. CBC with peripheral film, PT, APTT and Fibrinogen levels were done initially. Platelet aggregation studies, flow cytometry, and mutation analysis was done by Sanger sequencing. Genomic DNA was extracted from peripheral blood by QIAamp DNA Blood mini kit (Qiagen) and Exon specific PCR was done for GT gene and Direct gene sequenced on automated ABI-3130 Genetic Analyzer (Applied Biosystems). For any variation wild type was matched on HGMD (Human Gene Mutation Database http://www.hgmd.cf.ac.uk/ac/index.php) and wild type color fasta sequence (http://pga.gs.washington.edu/). Pathogenecity score was evaluated by using software tools including :
Polyphen-2(http://genetics.bwh.harvard.edu/pph2/) ,SNP&GO(http://snps.biofold.org/snps-and-go/index.html), MUpro (http://mupro.proteomics.ics.uci.edu/) ,SIFT (http://sift.jcvi.org), Provean (http://provean.jcvi.org/about.php) .
All samples were sequenced at the Gene Sequencing Lab of NIBD (National Institute of Blood diseases and Bone Marrow Transplantation) Karachi.
Results:
Mutations were identified in all patients. Missense mutations were seen in most of the GT patients. The remaining mutations were heterogeneous and were distributed throughout the length of the gene.
Conclusions:
The severe type I GT was the most common subtype found in this study.Carrier detection and genetic counseling in these families is a potentially effective alternative for decreasing the burden of severe type of GT.

Disclosures
No relevant conflicts of interest to declare.
A Predominant Mutation in Regulatory Region of SERPINC1 Gene and Venous Thrombosis
Authors: Hu Bei , Wang Qingyun , Tang Liang , Hu Yu , Liu Han 
Published: October 2019

Abstract
Background
Antithrombin (AT) is a critical physiological anticoagulant in haemostasis. SERPINC1, the gene encoding antithrombin, is 13.5kb in length and located on chromosome 1q23-25. Mutations in the SERPINC1 gene may cause antithrombin deficiency and are supposed to confer a high risk for venous thrombosis. However, most mutations reported so far are located in the coding or splicing region of this gene. Therefore, we investigate SERPINC1 gene to identify possible regulatory variants that would increase susceptibility of thrombosis in Chinese people.
Methods and Results
The potential promoter and 5'-untranslated region of SERPINC1 was screened by direct DNA sequencing in 100 individuals diagnosed with unprovoked venous thrombosis. Further genotyping was performed by Restriction Fragment Length Polymorphism (RFLP) analysis in case-control populations (1750 vs 1900). By DNA sequencing, we identified a c.-11.G>A transition, located 11bp upstream from the transcription initiation codon. Next case-control study showed that this mutation was present in ten individuals of thrombosis group and two of control group. All the mutation carriers we identified were heterozygotes. Therefore, individuals carrying the mutant A allele conferred a 5.42-fold increased risk for venous thrombosis (p<0.05) .
Conclusion
The c.-11G>A mutation of SERPINC1 gene is a risk factor for venous thrombosis in the Chinese population. It's the first time that a predominant mutation in the regulatory region of SERPINC1 gene reported to be associated with venous thrombosis in a large population.

Disclosures
No relevant conflicts of interest to declare.
Atherosclerotic Cardiovascular Risk in Adult Patients with Hemophilia: What We Can Do to Reduce Risk of Cardiovascular Events? Implementation of 2013 ACC/AHA Guideline on the Treatment of Blood Cholesterol in Patients with Hemophilia
Authors: Cisak Kamila Izabela , Pan Jianmin , Rai Shesh Nath , Ashby Patricia , Sharma Vivek R. 
Published: October 2019

Abstract
Introduction
Hemophilia A and B are genetic disorders characterized by deficiency of clotting factors resulting in delayed bleeding. Despite hypocoagulable state, patients with hemophilia are prone to developing coronary artery disease or its equivalents. It is known that proper treatment of dyslipidemia has relevant impact of atherosclerotic cardiovascular events reduction. The goal of our study was to determine implementation of newest 2013 ACC/AHA Guideline on the Treatment of Blood Cholesterol in our patients with hemophilia and assess how many more patients currently may require lipid-lowering therapy.
Methods
We performed retrospective chart review of patients followed at single hemophilia treatment center in United States. We included 30 patients with factor VIII or IX deficiency, age 30 and older, followed in clinic between 2005 and 2014 with available lipid profile results. Patients with acquired hemophilia were excluded from study. We used stepwise approach proposed by above guidelines and divided patients into four groups.
Results
4 patients among 30 were already on lipid lowering therapy. 1 (3.3%) additional patient [95% CI 0.001-0.17] required lipid lowering therapy due to presence of clinical atherosclerotic cardiovascular disease (group 1), 0 patients had LDL-C at least 190 mg/dl (group 2), 2 (6.7%) additional patients [95% CI 0.008-0.21] required therapy due to presence of diabetes mellitus and 40 to 75 year of age and LDL-C levels of 70 to 189 mg/dl (group 3); 9 (30%) additional patients [95% CI 0.17-0.51] should receive therapy due to age 40 to 75 and estimated 10-year ASCVD risk above 7.5%. We had total 12 (40%) additional patients among 30 with known lipid profile who were not on lipid lowering therapy but who require such therapy based on the latest 2013 ACC/AHA Guideline on the Treatment of Blood Cholesterol.
Conclusion
Aggressive cardiovascular risk factor modifications play a significant role in prevention of coronary artery disease, stroke and peripheral vascular disease. This may be even more relevant in patients with hemophilia who have an increased baseline risk of bleeding and may therefore be at greater risk of complications from anti-thrombotic therapies used for treating cardiovascular disease. Above results suggest that according to actual 2013 ACC/AHA Guideline on the Treatment of Blood Cholesterol, a significant number of patients with hemophilia may require lipid lowering therapy. It is important for hemophilia treatment centers to screen their patients with regard to this since many of them may either not have primary care physicians or may not be perceived as having high risk for cardiovascular disease due to their bleeding disorder.

Disclosures
No relevant conflicts of interest to declare.
Baseline Thrombin Generation Markers and Functional Antithrombin Levels in Sepsis Associated Coagulopathies Are Predictive of the Severity of Pathogenesis
Authors: Hoppensteadt Debra , Williams Mark , Syed Daneyal , Guler Nil , Mosier Michael , Fareed Jawed 
Published: October 2019

Abstract
Introduction: Severe sepsis remains the most common cause of death in critically ill patients. Thrombin plays a crucial role in the pathogenesis of sepsis associated disseminated intravascular coagulation (DIC). Thrombin generation markers and functional AT levels provide useful prognostic information in the management of these patients. The purpose of this study was to compare the baseline values of such thrombin generation markers of prothrombin fragment (F1.2), thrombin antithrombin complex (TAT) and functional antithrombin levels in patients enrolled in the Phase 2B and a Phase III study to investigate the therapeutic effect of recombinant thrombomodulin (ART-123) in sepsis associated coagulopathy.
Materials and Methods: Plasma samples from seven hundred and fifty eight patients enrolled in the ART-123 study, a phase 2b, international, multicenter, randomized controlled trial, and patients enrolled in an ongoing (n=200) phase III clinical trial were profiled for F1.2 and TAT using commercially available ELISA methods (Siemens, Deerfield, IL, USA). Functional AT activity was measured using a chromogenic substrate method. All of these samples included blood drawn prior to the first of ART-123 or placebo treatment.
Results: The results are tabulated in the following tables.
Phase 2b Study (n=750) Table 1.ParameterMeanStd ErrorMinMaxMedianQ1Q3F1.2 (pmol/L)515.541.7109,239305182571.5TAT (ng/ml)26.74.50.31,213.87.54.215.1AT (% NHP)84.91.383.020672.882.3101.3
Phase 3 Study (n=176) Table 2.ParameterMeanStd ErrorMinMaxMedianQ1Q3F1.2 (pmol/L)604.259.418.88,618401.8207.6695TAT (ng/ml)28.94.21.86524.812.46.1227.6AT (% NHP)51.424.82.1122.650.332.567.3
Discussion: The inclusion criteria for the phase 2b study was based on the revised ISTH criteria and the phase III study used an INR>1.4, 30K=<PLT<150K, shock, and/or respiratory organ dysfunction as criteria at baseline. The median values were TAT (7.5 ng/ml Phase 2b study; 12.4 ng/ml Phase III), F1.2 ( 305 pmol/L Phase 2b; 401.8 pmol/L Phase III) and AT ( 72.8 %, Phase 2b; 50.3% ,Phase III). There was a difference noted in the markers of thrombin generation between the two studies. The TAT and F1.2 were higher and the AT was lower in the Phase III study suggesting a higher generation of thrombin in this study. Therefore the strict criteria used in the Phase III study shows that this patient group had more activation of coagulation compared to the Phase 2b patients. Furthermore the modified criterion used for the recruitment of patients in the phase III study provides a defined project population to investigate the therapeutic effects of ART-123. These observations also underscore the importance of inclusion criterion for sepsis associated coagulopathy patients to validate the therapeutic effects of newer drugs such as ART-123.

Disclosures
Williams: Asahi Kasei Pharma America: Employment.
A Generic Version of Recombinant FVIIa Is Similar to the Branded Product (NovoSeven)
Authors: Sadeghi Nasir , O'Malley Paul , Kahn Daniel , Hoppensteadt Debra , Fareed Jawed 
Published: October 2019

Abstract
Background: Commercially available recombinant factor VIIa (Novoseven) is widely used in the management of hemophilia patients with inhibitors. Recently several generic versions of recombinant VIIa (rFVIIa) have become available. The generic versions of rFVIIa are claimed to be biosimilar to the barnded Novoseven (Novo Nordisk, Copenhagen, Denmark). The purpose of this study is to compare the US and European Novoseven products with a generic version of rFVIIa namely, Aryoseven (Aryogen, Tehran, Iran).
Methods: Four commercially available random lots of Novoseven were obtained from the US and European sources. Four different batches of Aryoseven were obtained from Aryogen. All individual rFVIIa preparations were diluted to obtain working concentrations of 100, 10, 1 and 0.1 ug/ml. Protein content (Lowry's method), molecular profile using surface enhanced laser desorption ionization (SELDI), gel electrophoretic profile (GEP), factor VII related antigen level (FVII:Ag), factor VII correction studies in depleted plasma and thrombin generation (TG) studies were carried out. In addition, VIIa/tissue factor mediated thrombin generation studies were carried out in various prothrombin complex concentrates such as Beriplex and Prothromplex.
Results: The protein content and SELDI mass spectrophotometric profile of all 4 rFVIIa preparations were comparable. There was no differences in the Novoseven obtained from the US and European sources. The GEP of the two groups of agents showed a comparable profile with distinct peaks at 50 KDa and 25 KDa. The FVII related antigen levels were also comparable in the Novoseven and Aryoseven preparations. Supplementation of both the Novoseven and Aryoseven preparations at 10 and 100 ug/ml resulted in a comparable correction of the factor VII deficient plasma as measured by PT(INR). Thrombin generation was comparable in the branded and generic product.
Conclusions: These results demonstrate that the US and European Aryoseven are comparable. Four batches of Novoseven and 4 individual clinical batches of Aryoseven were found to be comparable. When the US purchased Novoseven preparation was compared with the European Novoseven product, no differences were noted. Thus, the generic Aryoseven is biosimilar to barnded Novoseven and warrant in vivo validation studies.

Disclosures
No relevant conflicts of interest to declare.
Transformed Phenotype of Bone Marrow Stromal Cell Lines Derived from K14E7 Fancd2-/- mice
Authors: Sivanathan Aranee , Zhang Xichen , Franicola Darcy , Cao Shaonan , Shields Donna , Epperly Michael W , Greenberger Joel S. 
Published: October 2019

Abstract
To determine whether cytokeratin 14 promoter linked expression of the Human Papilloma Virus (HPV) oncogene detectably influenced biologic parameters of cell phenotypes other than squamous epithelium, continuous bone marrow cultures were derived from K14E7 Fancd2-/- mice (Park, et al., Cancer Research, 70(23): 9959-9968, 2010). Long-term bone marrow cultures derived from K14E7 Fancd2-/-, control Fancd2-/- (129/Sv), K14E7 (FVB/N), and wild type 129/Sv X FVB/N F1 mice were evaluated for longevity of hematopoiesis in long-term cultures and stromal cell lines were derived from each. Similar to Fancd2-/- mouse, long-term marrow cultures, K14E7 Fancd2-/- marrow cultures demonstrated decreased longevity of hematopoiesis with cessation of production of multi-lineage colony forming progenitor cells after 14 weeks. In contrast, wild type F1 and K14E7 long-term marrow cultures continued to produce hematopoietic cells for a significantly longer duration 25 weeks (p=0.0257). Bone marrow stromal and IL-3 dependent hematopoietic cell lines were derived from each genotype marrow culture. K14E7 Fancd2-/- hematopoietic cells showed reversal of the radiation resistance of Fancd2-/- IL-3 dependent cell lines (D0 of 1.34 ± 0.197Gy, ñ 4.0 ± 0.9 compared to D02.213 ± 0.124 Gy (p = 0.0284), ñ 3.3 ± 0.8. Thus, one phenotypic difference associated with K14E7 oncogene expression was reversal of radioresistance of Fancd2-/- hematopoietic cells. In contrast, bone marrow stromal cell lines from K14E7 Fancd2-/- remained radiosensitive similar to those from Fancd2-/- mice. K14E7 and wild type F1 marrow stromal cell lines showed intermediate radioresistance (p= 0.1759). To determine whether the E6/E7 oncogene had a biological effect in tissues other than squamous epithelium, tissue analysis for cytokeratins 13, 14, 6, and 10 was carried out. Cytokeratin 14 was detected only in squamous cells of the esophagus and oral cavity, not in bone marrow. E6/E7 oncogene was detected only in squamous cell lines expressing cytokeratin 14. However, bone marrow stromal cell lines from K14E7 Fancd2-/- marrow cultures demonstrated a unique phenomenon of cellular density, piling up and formation of tumors in vitro. Each of 22 single cell derived clonal sub-lines of K14E7 Fancd2-/- stromal cell lines demonstrated the same transformed phenotype. These data provide support for indirect effects of the E7 oncogene linked to the K14 promoter in Fancd2-/- hematopoietic and mesenchymal stem cell tissues.
Supported by research grant NIAID/NIH, U19A168021.

Disclosures
No relevant conflicts of interest to declare.