bluuebunny/crossref_metadata_2025_split huggingface dataset:
Detection of SNPs of Dck and Cda Genes in Five Hematological Malignant Tumor Cell Lines
Authors: Xiaoping Zhang , Jian Ziying , Chen Bao-An , Xu Peipei , Peng Miaoxin , Xia Guohua , Cheng Lu , Lu Zuhong 
Published: October 2019

Abstract
Abstract 5129

Background & Aim:
This study detected single nucleotide polymorphisms (SNPs) of deoxycytidine kinase(DCK) gene, cytidine deaminase(CDA) gene in five hematological malignant tumor cell lines, to find connections between sNPs and Cytarabine dose selection in the treatment of some hematopathy.

Materials & Methods:
Cell line K562, Ka, HL-60, U937, and Raji were cultured. The genomic DNA was isolated by QIAamp DNA Blood Mini kit. Designing primes were amplified by PCR to get the related DNA fragments. DCK gene A674G(rs111454937) C1644T(rs72552079), CDA gene A79C(rs2072671) G208A(rs60369023) was genotyped by means of matrix assisted laser desorption ionisation-time of flight mass spectrometry method (MALDI-TOF MS).

Results:
The genotype of locus A674G(rs111454937) on DCK gene in all five hematology system cell lines was AA; The genotype of locus C1644T(rs72552079) on DCK gene in all five hematology system cell lines was CC. The genotype of locus G208A(rs60369023) on CDA gene in all five hematology system cell lines was GG; For HL-60, U937, and Raji cell lines, the genotype of locus A79C(rs2072671) on CDA gene was AA, while for K562 and Ka cell lines, that was C/A.

Conclusions:
The genotype of locus A79C(rs2072671) on CDA gene was not all the same in the detected hematological malignant tumor cell lines, while the genotypes of the other 3 loci were same in the 5 cell lines.

Disclosures:
No relevant conflicts of interest to declare.
Tissue Factor and Hematological Neoplasias
Authors: Cesarman-Maus Gabriela N. , Braggio Esteban , Lome Carmen , Morales-Leyte Ana Lilia , Fonseca Rafael 
Published: October 2019

Abstract
Abstract 5132
There is a well- recognized correlation between cancer and aberrant hemostasis. Venous thromboembolism (VTE) in individuals with solid epithelial tumors has been associated with a poor prognosis, with more than three-fold higher risk of early death as compared to cancer patients without thrombosis. The expression of tissue factor (TF), a cell-surface membrane glycoprotein that triggers the activation of coagulation by cancer cells, is one of the main underlying mechanisms linking thrombosis and aggressive tumor behavior. TF is expressed in a variety of solid tumors in association with genetic events affecting oncogenes and tumor suppressor genes. However, the mechanisms of thrombosis in individuals with hematological malignancies may differ from those with solid tumors. We have previously shown that despite the high rate of thrombosis in multiple myeloma, malignant plasma cells only rarely express TF. We sought to determine TF gene (F3) and protein expression in hematological neoplasias. F3 expression profiling was studied on a variety of cell lines established from lymphoid and myeloid neoplasias available at Glaxo Smith Kline (GSK) Cancer Cell Line Genomic Profiling Dataset (https://array.nci.nih.gov/caarray/project/woost-00041). Interestingly, F3 expression was absent in all lymphoid neoplasias studied, in sharp contrast to acute myeloid leukemias (AML) and solid tumors, of which 30 and 90% expressed F3, respectively. Immunohistochemistry (IHC) confirmed the absence of TF protein expression in all indolent and high-grade B-cell lymphomas (99 patients, including germinal center and activated B-cell phenotype diffuse large B-cell lymphomas) and in all T-cell lymphomas/leukemias (20 patients) studied. IHC for TF was also negative in AML (11 patients) but positive in representative solid tumors (breast, pancreas, prostate), except for renal cell carcinoma which has been previously shown to lack TF. We propose that the pathogenesis of VTE associated with hematological neoplasias differs from that of solid tumors. Though TF from non-neoplastic cell sources may still be important for the prothrombotic state often seen in these patients, we show there is no evidence for a role of tumor-derived TF in the development of DVT, nor in neoplastic behavior. Thus, treatments directed against TF may not impact on prognosis in lymphoid, and non TF-expressing myeloid neoplasias.

Disclosures:
No relevant conflicts of interest to declare.
Hemostatic Factors and Shortened Sleep
Authors: Koktas Zehra , Vida Loyda , De Chavez Peter John D , Carnethon Mercedes , Green David 
Published: October 2019

Abstract
Abstract 5137

Introduction:
Enhanced coagulation has been reported in persons with sleep disturbances and sleep apnea. However, it is unclear whether shortened sleep duration in the absence of disordered breathing is associated with higher levels of hemostatic factors.

Methods:
In this cross-sectional study, 582 community-dwelling adults were randomly sampled. They underwent polysomnography to determine the apnea hypopnea index (AHI), a measure of sleep disordered breathing, and wrist actigraphy for 7 nights to assess sleep duration. A morning blood sample was collected for measurements of factor VIII (FVIII), von Willebrand Factor (vWF), thrombin antithrombin complexes (TAT), and plasminogen activator inhibitor-1 (PAI-1). Logistic regression analyses were used to model the odds of being in the upper quintile of each hemostatic factor by AHI score or sleep duration.

Results:
Participant ages ranged from 35 to 65 (mean 48), with 57% women and 30% Blacks. Hypertension was present in 18% and diabetes in 6%. The mean (SD) sleep duration was 7 (1. 07) hours/day, and 34% had an AHI ≥5. Mean values (SD) for hemostatic factors were FVIII (U/ml), 1. 06 (0. 41); vWF (U/ml), 1. 21 (0. 51); TAT (ng/ml), 2. 66 (2. 25); and PAI-1(ng/ml), 23. 4 (21. 4). vWF increased with age and FVIII, vWF, and TAT were higher in Blacks vs Whites. Associations (odds ratio, 95% confidence interval) were observed for FVIII with diabetes (3. 16, 1. 36–7. 32) and PAI-1 with hypertension (1. 9, 1. 06–3. 41) and diabetes (2. 58, 1. 15–5. 76) after adjustment for demographic and cardiovascular covariates.
In unadjusted analyses, participants with AHI ≥5 had elevated levels of FVIII and vWF, but this association was attenuated after adjustment for demographic and cardiovascular covariates (see Table). However, AHI ≥5 remained significantly associated with elevated PAI-1 in fully adjusted models. The majority of participants (90%) had a low likelihood of apnea (AHI <15), and their sleep duration showed no associations with hemostatic factors in any model.

Conclusion:
In a random population sample, we confirmed that mild apnea is associated with elevated PAI-1. However, among those persons free of apnea, there were no associations of sleep duration with FVIII, vWF, TAT or PAI-1.

Disclosures:
No relevant conflicts of interest to declare.
Elevated Plasma Levels of Soluble Platelet Glycoprotein VI (GPVI) in Patients with Thrombotic Patients
Authors: Wada Hideo , Nobori Tsutomu , Naitoh Katsuki 
Published: October 2019

Abstract
Abstract 5145

Introduction
Thrombotic microangiopathy (TMA) is caused by various conditions, such as decreased a ADAMTS13 level, activated or injured vascular endothelial cells or activated platelets and it must be diagnosed at the early stage. This study, examined the activation of platelets by measuring of soluble platelet glycoprotein VI (sGPVI) level in patients with TMA.

Materials and Methods
The plasma levels of sGPVI, ADAMTS13 activity, von Willebrand factor (VWF) and VWF propeptide (VWFpp) were measured in 40 healthy volunteers, 46 patients without thrombosis (TH), 15 postoperative patients, 13 with disseminated intravascular coagulation (DIC) and 70 with TMA. The plasma levels of GPVI and VWFpp were measured by ELISA and ADAMTS13 was measured by the FRETS assay. There were 27 TMA with markedly decreased ADAMTS13 levels (TMA-ADAMTS13) and 43 TMA without (TMA-Other).

Results
The plasma levels of sGPVI (median; 25. 0–75. 0%tile) were significantly higher in patients without TH (16. 2 ng/mL 12. 6–22. 5 ng/mL), postoperative patients (31. 6 ng/mL; 28. 3–35. 1 ng/mL), DIC (44. 5 ng/mL; 36. 6– 60. 8 ng/mL), TMA (40. 8 ng/mL; 32. 9–56. 7 ng/mL) than in healthy volunteers (11. 4 ng/mL; 9. 1–14. 8 ng/mL). In addition, the plasma levels of sGPVI post in postoperative patients were significantly higher than in patients without TH (p< 0. 001), and these levels were also significantly higher in those with TMA and DIC than in without TH (p< 0. 001, respectively). The plasma levels of sGPVI were significantly higher in patients with TTP-ADAMTS13 (36. 8 ng/mL; 30. 5–49. 0 ng/mL) and TMA-Other (51. 9 ng/mL; 37. 4–66. 3 ng/mL) than in patients without TH (p< 0. 001, respectively).

Conclusion
The measurement of sGPVI is therefore considered to be important for the diagnosis of TMA.

Disclosures:
Wada: Mochida pharmaceutical CO., LTD: Suporting GPVI assay Other.
Arsenic Trioxide Induces Platelet Apoptosis
Authors: Wu Yicun , Dai Jin , Zhang Weilin , Yan Rong , Ruan Changgeng , Dai Kesheng 
Published: October 2019

Abstract
Abstract 5149

Objective:
Arsenic trioxide, a component of Traditional Chinese Medicine, is known as an effective anticancer drug especially in the treatment of acute promyelocytic leukaemia (APL). APL has emerged as the most curable subtype of acute myeloid leukaemia since the widely use of arsenic trioxide-based chemotherapy. However, recent researches show that thrombocytopenia occurred in 79% of the relapsed or refractory APL patients treated with arsenic trioxide, and part of the APL patients had to be stopped treatment because of catastrophic bleeding, such as intracranial and pulmonary haemorrhage. Thrombocytopenia also occurred in 43% of the myelodysplastic syndrome patients treated with arsenic trioxide. Recently, arsenic trioxide has been proved to have a pro-apoptotic effect on various kinds of nucleated tumour cells or non-tumour cells. The effect of arsenic trioxide on enucleated platelet, however, still remains unclear. The aim of current study is to investigate whether arsenic trioxide induces platelet apoptosis.

Methods:
Washed platelets (3 × 108/ml) were incubated with different concentrations of arsenic trioxide or vehicle at 37°C for 4 hours. Then, mitochondrial inner transmembrane potential (ΔΨm) and phosphatidylserine (PS) exposure were tested by flow cytometry. In the mean time, the treated platelets were analyzed by western blot for the expression levels of pro-apoptotic protein (Bax), and anti-apoptotic proteins (Bcl-2 and Bcl-XL). Activation of caspase-3 was also examined by western blot using an anti-caspase-3 antibody.

Results:
ΔΨm depolarization and PS exposure were dose-dependently induced in platelets incubated with different concentrations (2 uM, 4 uM, 8 uM, 16 uM) of arsenic trioxide as detected by flow cytometry, and the lowest concentration of arsenic trioxide incurring ΔΨm depolarization and PS exposure was 4 umol/L. Simultaneously, arsenic trioxide induced up-regulation of Bax and down-regulation of Bcl-2 and Bcl-XL in a dose-dependent manner. Furthermore, 17 kD cleaved caspase-3 fragments were dose-dependently induced in platelets treated with different concentrations of arsenic trioxide indicating that caspase-3 was activated by arsenic trioxide.

Conclusions:
Taken together, the data indicate that arsenic trioxide induces platelet apoptosis in vitro, which might suggest a novel pathogenic mechanism of thrombocytopenia in the patients who treated with arsenic trioxide.

Disclosures:
No relevant conflicts of interest to declare.
Red Cell Microparticles (RMP) and Other Cell-Derived Microparticles (C-MP) in Hemolytic Anemias
Authors: Johansen Max E , Jy Wenche , Dudkiewicz Pamela B , Ahn Yeon S 
Published: October 2019

Abstract
Abstract 5158

Introduction:
The lifespan of red cells (RBC) are shortened by several mechanisms in hemolytic anemias (HA): Ab-mediated hemolysis in autoimmune hemolytic anemia (AIHA), complement-mediated lysis in paroxysmal nocturnal hemoglobinuria (PNH), microangiopathy in thrombotic thrombocytopenic purpura (TTP) and hemoglobinopathy in sickle cell anemia (SS) and thalassemia (Thal). During hemolysis, RMP are released along with MP from platelets (PMP), endothelia (EMP) and leukocytes (LMP). The role of MP in HA is unknown but may be involved in complications of HA.

Methods:
We investigated RMP and other MP profiles in the following hemolytic anemias in active phase: 14 patients with AIHA, 10 with TTP, 4 with PNH, 8 with SS or Thalassemia (SS - Thal), along with 60 healthy controls. Whole, citrated blood was centrifuged at 1600xgfor 10 minutes to yield platelet-poor plasma (PPP). Microparticles in the PPP were measured by FITC- or PE-conjugated mAb specific to the aforementioned cell types. Control plasma was prepared in an identical fashion from healthy volunteers. We compared RMP profiles and other MP in HA during active phase and in remission. Associations with clinical features were calculated using the R statistical software.

Results:
Means and standard deviations of C-MP in the different types of HA are shown in the TABLE. Mean RMP and PMP were highest in SS-Thal, followed by AIHA. Mean Annexin V binding procoagluant MP were higher in all subgroups of HA and highest in SS-Thal, followed by TTP. EMP in all subgroups was similar or lower than controls.
Regarding clinical associations, high RMP levels were associated with low hemoglobin (p=0. 01), high reticulocytes (p=0. 003), and high LDH (p=0. 01) across all groups. High reticuloctye counts (>3. 0 %) were also associated with significantly elevated PMP (p=0. 04) and total MP counts by Ulex (p=0. 003). HAs in remission had marginally lower RMP levels than controls (p=0. 05). None of the subtypes had significantly higher EMP, LMP, or PMP compared to each other or controls.

Conclusion:
The small population yielded large standard deviations in the MP counts and did not allow statistical study among subgroups of patients with HA. However prothrombotic MPs such as Annexin V, PMP tend to be higher in HA. A larger scale future study will clarify this issue. The constellation of elevated LDH, reticulocytes, and decreased hemoglobin were associated with increased RMP. Therefore we suggest that elevated RMP may be viewed as an additional, new diagnostic marker of active hemolysis.

Disclosures:
No relevant conflicts of interest to declare.
Diagnosis of Auto Immune Hemolytic Anemia by Detection of RBC-Bound IgG Antibodies, Using Flow-Cytometry
Authors: Khungar Jitendra Mohan , Pati Hara Prasad Prasad , Mahapatra Manoranjan 
Published: October 2019

Abstract
Abstract 5159

Introduction:
Auto Immune Hemolytic Anemia (AIHA) is one of the most common types of acquired hemolytic anaemias. Its main cause is auto antibody mediated rapid destruction of RBCs. Detection of these autoantibodies to erythrocytes is of fundamental importance for diagnosis. A number of methodologies have been tried for detection & evaluation of these autoantibodies. Demonstration of a positive Direct Antiglobulin Test (DAT) against these autoantibodies is an important serological assay in the diagnosis of auto immune hemolytic anemia (AIHA). This test is also considered as pathognomonic of immune-mediated hemolysis. This routinely used direct antiglobulin test (DAT) has the disadvantage of low sensitivity and does not detect low levels of red cell auto antibodies leading sometimes to false negative results. Flow cytometry can effectively diagnose such patients of auto immune hemolytic anemia with low levels of autoantibodies. Role of flow cytometry in the diagnosis of several non-malignant haematological disorders is being explored & present study has been conducted with the same objective.

Aims & Objectives:
This study was conducted with the following aims and objectives:
•To assess the utility of flow-cytometry (FCM) in the diagnosis of suspected AIHA patients.
•Compare the sensitivity of flow-cytometry (FCM) with Direct Antiglobulin Test (DAT) by Gel-card Test (GT).
•To assess the positivity in DAT negative cases by flow-cytometry in suspected AIHA cases.

Material & Methods:
This was a prospective study, carried out in Haematology Deptt of All India Institute of Medical Sciences, where patients with suspected auto immune hemolytic anemia (AIHA) were studied during two years period. Blood samples of suspected patients of AIHA were tested by Gel Card Test as well as by Flow cytometry for detection of RBC bound IgG.

Results:
A total of 50 patients with suspected diagnosis of auto immune hemolytic anemia (AIHA) were studied by flow-cytometry as well as by Gel card test (GT) for detection of RBC bound IgG. Out of these 50 cases, 41 cases have turned out to be positive and 9 were negative by flow-cytometry. The quantification of positivity by flow-cytometry was obtained by calculating percentage fluorescence. The same 50 cases were also tested by Gel card test (GT). By Gel card test, out of 50 cases, 34 were positive & 16 were negative. Therefore, there were 7 cases which were negative for RBC bound IgG by Gel card test and these were positive by flow-cytometry.
The flow cytometry figures of these cases will be shown & discussed in the presentation.

Conclusions:
Flow-cytometry is a reliable and sensitive method of detecting RBC-bound IgG antibodies for the diagnosis of auto immune hemolytic anemia. This can be used as a new routine diagnostic technique for auto immune hemolytic anemia.

Disclosures:
No relevant conflicts of interest to declare.
Efficacy and Safety of Monoclonal Anti-CD20 Antibody Rituximab Combined Cyclophosphamide in Treatment of Refractory and Recurrent Autoimmune Hemolytic Anemia
Authors: Shao Zonghong , Liu Hong , Xing Limin , Wu Yuhong , Qu Wen , Liang Yong , Guan Jing , Wang Guojin , Wang Xiaoming , Song Jia , Li Lijuan , Wang Huaquan 
Published: October 2019

Abstract
Abstract 5161

Objectives:
To better assess the efficacy and safety of monoclonal anti-CD20 antibody rituximab in the treatment of refractory and recurrent autoimmune hemolytic anemia.

Patients and methods:
7 cases with autoimmune hemolytic anemia (including 1 case of Evans syndrome) were enrolled into this study, they were treated with rituximab (375 mg/m2, once per week, 2–6 times) and Cyclophosphamide(1g/10days, 2–6 times) combined with intravenous immunoglobulin (IVIG) (10g/week, given 1 day after rituximab treatment).

Results:
All 7 patients showed good responses. 6 achieved complete remission (CR) and 1 achieved partial remission (PR). Responses were seen at 1th to 10th month after the first dose of rituximab and the mean response time was approximately 2. 5 months. Average follow-up time for them isfor the patients was 27 months. All patients remained in remission at the 12-month follow-up. At the time of 24-month follow up, 3 patients showed elevated indirect bilirubin and increased reticulocyte counts. One of the 3 patients achieved CR after additional rituximab therapy, and the other 2 PR after additional cyclophosphamide therapy. At the time of 36-month follow up, 1 patient relapsed and was retreated with 3 cycles of rituximab and eventually reached PR. All patients tolerated the treatment well with only mild side effects.

Conclusion:
rituximab is highly effective and relatively safe in patients with refractory and recurrent autoimmune hemolytic anemia. Maybe Additional treatment can be given in patients with relapse after 1–2 years.

Disclosures:
No relevant conflicts of interest to declare.
Alfa Thalassemia Heterozygous: How the New Information Provided by the Routine Hematology Analysers May Help in Its Differential Diagnosis or Flagging
Authors: Barella Susanna , Simon-Lopez Ramon , Gaetano Nicola Di , Galanello Renzo 
Published: October 2019

Abstract
Abstract 5183
Carriers of α°-thalassemia (α-thalassemia trait) show microcytosis, hypochromia, and normal percentages of HbA2 and HbF. Carriers of α + -thalassemia (α-thalassemia silent carrier) have either a silent hematologic phenotype or present with a moderate thalassemia-like hematologic picture. Homozygosity for α + -thalassemia results in an α°-thalassemia (α-thalassemia trait) hematologic phenotype.

Diagnostic of Alfa Thalassemia Heterozygous:
Classic testing for α-thalassemia includes: hematologic testing of red blood cell indices, peripheral blood smear, supravital stain to detect RBC inclusion bodies, and qualitative and quantitative hemoglobin analysis. HBA1, the gene encoding α1-globin, and HBA2, the gene encoding α2-globin, are the two genes most commonly associated with α-thalassemia. Molecular genetic testing of HBA1 and HBA2 detects deletions in about 90% and point mutations in about 10% of affected individuals.

Objective:
Recently have been developed new parameters and information in the new automated hematology analyzer called DxH8008™ from Beckman Coulter as @MSCV, @RSF, @MAF, @LHD% and many morphological parameters for RBC and Reticulocytes calles Cell Population Data. All this parameters may be used to create flagging for laboratory use only (LUO) or Research use only (RUO). The purpose of this study is to investigate the possible use or utility of this new information for the screening/flagging of Alfa Thalassemia Heterozygous.

Patient and Methods:
We have collected 48 patients with Alfa Thalassemia Heterozygous All of them were confirmed by red cell morphology, Hgb Electroforesis, cromatography in liquid phase in human whole blood for the determination of Hemoglobin A2, F, A1c, and identification of abnormal hemoglobins and DNA analysis (DNA Analysis by GAP-PCR). We have compared these patients with a control group (184 individuals) and with other anemias (see Table 1).

Results:
Using ROC analysis, the best parameters differentiating the Alfa Thalassemia Heterozygous from the normals were: MCV (AUC 0. 963), @RSF (AUC 0. 936), @MSCV (AUC 0. 912), RDW (AUC 0. 848), @MAF(AUC 0. 845), @LHD(AUC 0. 829).
Using ROC analysis, the best parameters differentiating the Alfa Thalassemia Heterozygous from other anemias (excluding normals) were: RDW (AUC 0. 885), @MAF(AUC 0. 857), @MCNRET (AUC 0. 832).

Disclosures:
Simon-Lopez: Beckman Coulter: @LHD, @MAF, @RSF, @LHD, @MAF, @RSF Patents & Royalties, Employment. Di Gaetano:Instrumentation Laboratory spa: Work for a distributor of Beckman Coulter Instruments in Italy Other. Galanello:Ferrokin: Research Funding; Apopharma: Research Funding, Speakers Bureau; Novartis: Research Funding, Speakers Bureau.