bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of a Histone Deacetylase Inhibitor On Myeloid Leukemia Cell Line with Mixed-Lineage Leukemia Partial Tandem Duplication
Authors: Keino Dai , Hayashi Mutsumi , Ohyama Ryo , Morimoto Mizuho , Kondoh Kensuke , Shimada Akira , Yamada Taketo , Kinoshita Akitoshi 
Published: October 2019

Abstract
Abstract 4331

BACKGROUND:
Mixed lineage leukemia-Partial tandem duplication (MLL-PTD) is one of the MLL gene rearrangements accompanying leukemia, and is associated with a worse prognosis. Since MLL regulates a gene transcription, histone deacetylase inhibitors (HDACi) have been considered to exert antitumor effects through epigenetic modification. We established a leukemia cell line KOPM-88 containing MLL-PTD from a child with acute myeloid leukemia (AML). Non-obese diabetic severe combined immunodeficient (NOD/SCID) mice inoculated with KOPM-88 cells exhibited leukemic infiltrations in the bone marrow and hemiparalysis because of compression myelopathy (Hayashi et al., Br J Haematol, 2007). We therefore investigated the effects of HDACi on KOPM-88 cells and NOD/SCID mice inoculated with KOPM-88 cells.

MATERIALS & METHODS:
The cell lines employed were KOPM-88 and K-562. We chose K-562 as a control because it contains a wild type MLL. Firstly, a HDACi, trichostatin A (TSA), was added with 100 nM/L concentration in KOPM-88 cells for 24 hours and in K-562 cells for 12 hours. Apoptosis was evaluated by using a flow cytometer. MLL-PTD mRNA expression was measured by using a real time PCR before and after incubation with TSA. Secondary, 5×106 of KOPM-88 cells were transplanted in NOD/SCID mice (n=32). Then, TSA was injected intraperitoneally into a half of the transplanted mice three times a week for 5 weeks (TSA group, n=16). DMSO, which was used as a solution of TSA, was injected intraperitoneally into the half rest of mice (control group, n=16). Hemiparalysis was counted as an event. The probability of event-free survival (EFS) and the probability of overall survival (OS) were analyzed using the Kaplan-Meier method.

RESULTS:
TSA significantly increased the apoptosis in KOPM-88 cells (30.03±2.83% vs 53.17±2.11%,p <0.01), but it did not significantly increased the apoptosis in K-562 cells (8.15±0.20% vs 14.04±0.23%,p=0.08). TSA also significantly decreased the level of MLL-PTD mRNA in KOPM-88 cells (1.78±0.35 copies/μgRNA vs 0.38±0.33 copies/μgRNA, p <0.05). The median follow-up of the TSA group was 178 days (range; 30–210), and that of the control group was 149 days (range; 98–210). Although seven months-EFS was significantly better in the TSA group than in the control group (78±0.8% vs 43.8±0.8%,p <0.05), seven months-OS was similar between the TSA group and the control group (43.8±0.8% vs 18.8±0.6%,p=0.25).

CONCLUSIONS:
Our study suggests that HDACi exerts an anti-leukemic effect partly from suppression of MLL-PTD transcription on leukemia cells with MLL-PTD. This anti-leukemic effect of HDACi was also confirmed in vivo mice model. The patients with MLL-PTD leukemia may benefit from HDACi treatment.

Disclosures:
No relevant conflicts of interest to declare.
Treatment of Acute Promyelocytic Leukemia (APL) At Tawam Hospital UAE
Authors: Alam Arif , Narayanan Harimohan , Sonday Shanaaz , Hussain Sabir , Lal Amar , Burney Mohammad , Kristensen Jorgen 
Published: October 2019

Abstract
Abstract 4347
Acute Promyelocytic Leukemia (APL) is a unique sub-type of Acute Myeloid leukemia associated with a balanced reciprocal translocation between chromosomes 15 and 17 and 80% of the cases present with bleeding diathesis caused by severe coagulopathy. The translocation generates a fusion transcript joining the PML(promyelocyte) and RAR-α (retinoic acid receptor-α) genes. The therapy of APL has been revolutionized by the introduction of differentiating agents All Trans Retinoic Acid (ATRA) and Arsenic Trioxide.
All patients were treated as per Pethema protocol. Initially LPA99 (Sanz MA. Blood. 1999 Nov 1;94(9):3015-21) and since January 2012 a risk adapted therapy based on LPA2005 (Sanz MA et al. Blood June 24, 2010 vol. 115 no. 25 5137–5146). Treatment included induction followed by 3 cycles of consolidation and two years of maintenance.
Nineteen patients were diagnosed with APL between January 2009 and June 2012. Three patients are excluded from the analysis as karyotyping and/or PCR did not confirm the diagnosis. The median age was 35 years (range 22–53 years). Male to female ratio was 4:1. Nine (56%) patients were stratified as high risk (WBC ≥ 10 ×109/l) while, seven (44%) as intermediate risk and low risk (WBC < 10 ×109/l). Three (19%) patients had early death despite treatment and supportive care. The cause of death was intracranial hemorrhage (1) pulmonary hemorrhage (1) and multi-organ failure (1). Thirteen patients achieved a complete morphological and molecular remission (80%). There has been only 1 case of treatment failure (high risk at presentation). This patient was successfully re-induced with arsenic trioxide and achieved second molecular complete remission. Unfortunately he relapsed a second time and is currently alive in third morphological remission but remains PCR positive for PML/RARα 33 months after diagnosis.
Our limited experience shows favorable outcome (CR 80%) for the treatment of APL using the Pethema protocol compared to published data (Tallman MS. Blood 2002;99(3):759–767). Early death rate remains high despite intensive supportive care. The only variable is the availability and initiation of ATRA at the clinical suspicion of diagnosis both at the referring hospitals and treatment center.

Disclosures:
No relevant conflicts of interest to declare.
Indirect Comparisons of the New Oral Anticoagulants in Patients with Non-Valvular Atrial Fibrillation
Authors: Zolfaghari Shabnam , Harenberg Job , Marx Svetlana , Wehling Martin 
Published: October 2019

Abstract
Abstract 4363
The efficacy and safety of new oral anticoagulants has been demonstrated for prevention of ischemic stroke and systemic embolism in patients with non-valvular atrial fibrillation (NVAF) by dabigatran in the RE-LY trial (150mg and 110mg bid), rivaroxaban in the ROCKET AF trial (20mg od), and apixaban in the ARISTOTLE trial (5mg bid) versus INR-adjusted warfarin. Direct comparisons of the NOACs in this indication are unlikely to be performed.
A total of 4 indirect comparisons of these trials on the efficacy and safety of NOACs in patients with NVAF have now been published within only 3 months (Lip et al 2012, Harenberg et al 2012, Mantha et al 2012, Wells et al 2012). Here, we compare the results of these 4 network meta-analysis (NMA).
In all 4 NMAs of the 3 new oral anticoagulants dabigatran (150mg bid) showed superior efficacy in preventing ischemic stroke plus systemic embolism to dabigatran (110mg bid, p<0.04) and rivaroxaban (p<0.04). Apixaban had equivalent efficacy with rivaroxaban and dabigatran (either dose). Apixaban was safer (less major bleeding) than dabigatran (150mg bid, p<0.04) or rivaroxaban (p<0.005). Intracerebral haemorrhage occurred with equal frequency for all agents and regimens except for rivaroxaban (higher risk than dabigatran 110mg bid, p<0.005). Myocardial infarction occurred less frequently with rivaroxaban and apixaban compared to either dose of dabigatran (all p<0.05). All-cause mortality was not different for any agent or regimen. Some minor differences between the NMAs may result from the approved doses of dabigatran by the FDA (150mg bid and 75mg bid) and EMA (150mg bid and 110mg bid), as the inclusion of the 110 mg bid dose of dabigatran into the NMA may not be seen relevant in the US.
Based on this comparison, doctors and patients have to decide which suggestions of the 4 groups of authors seem more convincing: to change to or to start with one of the NOACs depending on the individual thrombotic or bleeding risk or to wait for the results from a large (and expensive) head-to-head randomised controlled trial which may take years to perform.

Disclosures:
No relevant conflicts of interest to declare.
Bleeding Intensity Following Tooth Extraction Is Moderated by Anxiety Independently of Treatment with anticoagulants– Development of an Internet Based Questionnaire Across Countries
Authors: Zolfaghari Shabnam , Marx Svetlana , Fischer Jeelka , Hassani Parastoo , Damian Marinella , Frölich Lutz , Harenberg Job 
Published: October 2019

Abstract
Abstract 4364
Anticoagulation with dose adjusted vitamin-K antagonists (VKA, therapeutic range of the international normalized ratio (INR) of 2 to 3) and low dose aspirin (65 to 330 mg daily) enhance bleeding following tooth extraction. However, intensity of bleeding varies substantially between patients. Guidelines disagree regarding interruption or continuation of anticoagulation for tooth extraction. We hypothesized that personality traits may moderate bleeding intensity under anticoagulation.
A total of 180 patients (77 female, 103 male, 49.9+15.3 years (mean, standard deviation)) underwent tooth extraction without interruption of anticoagulation. Sixty three patients did not take any anticoagulant (group 1), 60 patients were on aspirin (group 2), and 57 patients on VKA (INR 2–3, group 3). Patients completed a validated state-trait anxiety inventory (STAI), a personality inventory with 12 dimensions (Freiburg personality inventory (FPI)), and a self-developed questionnaire on general attitudes regarding general feeling and anxiety before tooth extraction. Dentists (JF, PH) rated bleeding intensity ranging from 0 to 9 according a standardized protocol.
Intensity of bleeding after tooth extraction was higher in group 3 (score <5 in 40%, score >5 in 60% of patients) compared to groups 1 (40% and 60%) and 2 (53% and 47%) and higher in group 2 compared to group 1 (p<0.005, chi-square test) as expected. Higher anxiety values in STAI questionnaire correlated positively with higher bleeding scores (p<0.0001). High values of some of the FPI dimensions correlated positively with self-consciousness (p<0.0001), physical complaints (p<0.001), and emotionality (p<0.0001) life, and negatively with satisfaction (p<0.02) and extraversion (p<0.003) without differences between groups 1 to 3. Anxiety symptoms of the self questionnaire were identified as moderating factor on bleeding intensity following tooth extraction independently of anticoagulation (p<0.0001) according multinominal regression analysis. Other items of the self questionnaire such as physiological stress symptoms, regular performance of visits, bad experience with dentists, and pains during tooth extraction did not influence bleeding intensity.
A short questionnaire is developed to identify the anxiety score of patients as a tool for a non pharmaceutical medical intervention to reduce bleeding complications following tooth extraction. Because differences of the moderating effect of bleeding by personality traits may exist between cultures, the questionnaire will be made available across countries www.blutverduennung.uni-hd.de

Disclosures:
No relevant conflicts of interest to declare.
Use of recombinant factor VIIa in treatment of bleeding episodes for patients with refractoriness to platelet transfusion therapy
Authors: Sandilya Vijay , Gupta Namita 
Published: October 2019

Abstract
Abstract 4382
Recombinant factor VIIa (rVIIa) is currently used for treatment and prevention of bleeding episodes in hemophilia with inhibitors, congenital factor VII deficiency, and acquired hemophilia. We report a series of 3 cases of immune mediated refractoriness to platelet transfusion therapy that were successfully treated with rVIIa, an off-label use. Data was obtained retrospectively over a 6 month period. All patients were known to be platelet refractory with presence of significant titers of plasma HLA antibodies. Two patients had undergone matched unrelated donor HSCT and the third patient suffered from Myelodysplastic syndrome (MDS). Two of the patients presented with GI bleeding and the third with intracranial bleeding. Due to limited availability of HLA matched platelets and negligible increment in platelet count from ABO matched platelets, use of factor VIIa was considered. Transfusion with PRBCs to maintain a Hemoglobin (Hb) > 8.0g/dl and close monitoring of coagulation parameters were performed. Patients received boluses of rFVIIa at 90mcg/kg, every 3–12 hrs. The dose of rFVIIa ranged from 10800–28,000 mcg/day. All three episodes of bleeding achieved clinical or radiologic improvement. No toxicities or arteriothrombotic events were observed with rFVIIa use. Our experience indicates that rFVIIa might be an effective treatment option for patients with refractoriness to platelet transfusion therapy. The optimal dosing schedule in this clinical situation needs to be studied.
Patient demographics and outcomes Patient Diagnosis Pre-intervention Hb/platelet rFVIIa used (microgms) PRBC/Platelet transfused (units) Clinical outcome SV – GI bleed ALL-MUD, BMT, day +102. 7.9/4000 28000/d × 4 days 6/3 Bleeding stopped and Hemoglobin stabilized on day 4 CS- GI bleed AML-MUD,BMT, day + 78 7.8/3000 25200/d × 2 days 2/2 Bleeding stopped and Hemoglobin stabilized on day 2 MW-Intracranial hemorrhage MDS – IPSS risk group Intemediate-2 7.6/6000 10800/d × 2 days 2/2 Improvement in Neurological exam and stabilization on CT scan seen by day 3

Disclosures:
Off Label Use: Recombinant factor VIIa for refractoriness to platelet transfusion therapy.
The W–x–x–W Motif in the TSR1 of ADAMTS13 Is Important for Its Secretion
Authors: Ling Jing , Su Jian , Ma Zhenni , Ruan Changgeng 
Published: October 2019

Abstract
Abstract 4389

Introduction:
The W-x-x-W motif is commonly found in the thrombospondin type 1 repeat (TSR) of various extra cellular proteins called TSR super family proteins, including thrombospondins, ADAMTSs, F-spondin and properdin. The W–x–x–W motif is known to bind with heparin, heparan sulfate proteoglycans, collagen and transforming growth factor-β, suggesting functional significance in cell–cell interaction and/or cellular signaling. However, the function of W-x-x-W in ADAMTS13 is unclear. In this study, we investigated the role of the W-x-x-W motif of ADAMTS13.

Materials and Methods:
We generated a W-x-x-W mutant (W387A) construct of ADAMTS13, and expressed the mutant and the wild-type constructs in HELA cells. Percentage of the protein secretion was defined as the concentration in the culture medium divided by the concentrations in the culture medium and cell lysates, multiplied by 100%. The binding affinity of the mutant or wild-type ADAMTS13 was investigated by enzyme-linked immunosorbent assay. Measurement of ADAMTS13 proteolytic activity toward von Willebrand factor (VWF) multimers was based on the generation of a dimeric 176-kDa fragment resulting from cleavage of VWF at the Y1605-M1606 bond, under denaturing condition and high shear stress condition, analyzed by Western blots.

Results:
SDS-PAGE gel analysis showed that the W387A mutant was secreted less efficiently relative to the wild-type construct. As for the binding affinity for the VWF multimer, there was no difference between the wild-type and mutant ADAMTS13. The W387A mutant was less active under denaturing condition; the same result was reproduced when FRETS-VWF73 was used as the substrate. However, under high shear stress condition, the mutant was as efficient as the wild-type ADAMTS13.

Conclusions:
The W–x–x–W motif is necessary for efficient secretion of ADATMS13. Further studies are needed to determine the contribution of the motif to the VWF cleave activity of ADAMTS13.

Disclosures:
No relevant conflicts of interest to declare.
Comparison of the Use of TFPI Inhibition and Plasma Dilution to Increase the Sensitivity of Thrombin Generation Assay to Measure the Procoagulant Activity of Microvesicles
Authors: Gheldof Damien DG , Mullier François , Chatelain Bernard , Dogne jean-Michel , Chatelain Christian 
Published: October 2019

Abstract
Abstract 4392

Introduction:
Patients with cancer have a 7- to 10- fold increased risk of developing venous thromboembolism. Circulating microvesicles (MVs) could be a predictive biomarker for venous thromboembolism in cancer. Thrombin generation assay is a useful technique to detect procoagulant activity of MVs. However, thrombin generation assay suffers from a lack of sensitivity due to the presence of Tissue Factor Pathway Inhibitor (TFPI) in plasma.

Aims:
To improve the sensitivity of thrombin generation assay to tissue factor (TF) by limiting the interference of TFPI.

Methods:
Serial dilutions of MDA-MB231 cells were incubated for 45 min at 37°C to generate MVs. Samples were then centrifuged and supernatants which contain MVs were used for thrombin generation assay. Normal pooled plasma was incubated with inhibitor of TFPI or was diluted twice to decrease plasma level of TFPI. Lagtime was used as a surrogate marker of thrombin generation assay to detect procoagulant activity of MVs.

Results:
i) Inhibition of TFPI decreased twice the cell concentration needed for a significant reduction of lagtime and decreased 2.4-fold the intra-assay variability. ii) Plasma dilution had no impact on the thrombin generation assay sensitivity when thrombin generation assay was triggered by MVs derived from MDA-MB-231.

Conclusions:
Thrombin generation is a very sensitive method to study the procoagulant activity of TF-MVs. The sensitivity can be increased by inhibition of TFPI with specific monoclonal antibody against its Kunitz Domain I. A twice plasma dilution is an interesting alternative to study the procoagulant activity of MVs by thrombin generation assay with a good sensitivity, especially when low plasma quantities are available.

Disclosures:
No relevant conflicts of interest to declare.
Identification of Novel Fusion Genes with 28S Ribosomal DNA in Hematologic Malignancies
Authors: Kobayashi Satoru , Taki Tomohiko , Nagoshi Hisao , Chinen Yoshiaki , Yokokawa Yuichi , Kanegane Hirokazu , Matsumoto Yosuke , Kuroda Junya , Horiike Shigeo , Nishida Kazuhiro , Taniwaki Masafumi 
Published: October 2019

Abstract
Abstract 4418
Chromosomal translocations play a crucial role in tumorigenesis, often resulting in the formation of chimeric genes or in gene deregulation through position effects. Hematologic malignancies are associated with a large number of such rearrangements. In this study, we have identified 28S ribosomal DNA (RN28S1) gene as a novel fusion partner of the B-cell leukemia/lymphoma 11B (BCL11B), immunoglobulin kappa variable 3–20(IGKV3–20), and component of oligomeric golgi complex 1 (COG1) locus in T-cell acute lymphoblastic leukemia (T-ALL), Burkitt lymphoma (BL) and multiple myeloma (MM), respectively.
RN28S1-BCL11B fusion gene was identified in T-ALL with t(6;14)(q25;q32) by cDNA bubble PCR using BCL11B primers, whereas the breakpoint of 14q32 was near, but out of, the BCL11B. BLAST search revealed that the RN28S1 was fused to BCL11B within exon3 (coding region). During the expression analysis of RN28S1-BCL11B, We identified the same type of RN28S1-BCL11B fusion transcripts in a BL and 2 MM cell lines, and unexpectedly identified other RN28S1-fusion transcripts in hematologic malignancy cell lines. Fusion of RN28S1 and IGKV3–20 exon2 (coding region) was detected in 2 BL and a MM cell lines, and that of RN28S1 and COG1 within exon 13 (coding region) in a MM cell line. Furthermore, RN28S1 breakpoint was concentrated in between 1.4∼1.8kb.
RN28S1 is located in the p12 region of chromosomes 13, 14, 15, 21 and 22. The ribosomal RNA (rDNA) gene repeats are essential housekeeping genes found in all organisms. A gene amplification system maintains large clusters of tandemly repeated copies in the chromosome, and as the product, ribosomal RNA accounts for ∼70% of total RNA in the cell.
rDNA exists in high copy number in most eukaryotic cells. Repeatitive genes tend to lose copies through homologous recombination among the repeats. The rDNA has a gene amplification system, thought to be present in most eukaryotes, that function to keep high copy numbers. In the tumor cells, rDNA is transcribed higher than normal cells for cell proliferation. Thus, that may be associated with chromosome instability.
RN28S1 degradation was reported to be associated with apoptosis of hematologic malignancies, and previously, another group was detected RN28S1-BCL6 fusion genes in gastric lymphoma. This case was no expression of BCL6 protein. In conclusion, these results suggest that RN28S1 fusions are concerned with the play something role of tumorigenesis. Because of RN28S1 fused to BCL11B, IGKV3–20 and COG1 whose breakpoints were involved in the coding region, we speculate that these fusion genes are not produce normal structure.

Disclosures:
No relevant conflicts of interest to declare.
Micro-RNA Profile of Childhood Acute Lymphoblastic Leukemia
Authors: Karagun Barbaros Sahin , Antmen Bulent , Sasmaz Ilgen , Tanriverdi Kahraman , Ucar Gulsum , Kilinc Yurdanur 
Published: October 2019

Abstract
Abstract 4824
Micro-RNAs are functional, non-protein coding RNA molecules and their transcriptions provided by intron or exon regions of the genome and non-protein coding regions of RNA genes. The role of micro-RNAs in acute leukemia has become the subject of research increasingly. In this study we aimed to identify micro-RNA profiles in the childhood acute leukemia that one of hematologic malignancies.
Fourty nine patients who were diagnosed with acute leukemia and admitted to Cukurova University Faculty of Medicine Department of Pediatric Hematology between December 2010 and September 2011. Blood samples were taken twice in patient groups at diagnosis and during remission and plasma samples were stored. Blood samples were taken once in the healthy group and plasma were separated. The plasma samples were investigated by PCR analysis of micro-RNA. Acute leukemia was diagnosed by cytomorphological, immuno histochemical and flow cytometric studies. Thirtyone patients who were diagnosed with ALL and fortyseven healthy children as a control group were included to study.
miR20a, miR25, miR92a, miR30c, miR106b, miR203, miR150, miR192, miR302c, miR184, miR218, miR320, miR342-3p, miR223, miR328, miR483-5p, miR376a, miR381, miR451, miR576-3p, miR548a levels were increased in newly diagnosed ALL patients when compared to healthy controls (p <0.05). The miR20b, miR342-3p and miR548a levels were found higher in healthy controls than the newly diagnosed ALL patients group (p <0.05) Healthy control groups when compared with pediatric ALL patients whose in remission; miR769-3p, miR20a, miR92a, miR16, miR27b, miR192, miR320, miR223, miR484, miR451 levels were found higher in healthy control groups than the patients. Newly diagnosed pediatric ALL patients compared with patients whose in remission; miR30c, miR106b, miR25, miR184, miR218, miR302c, miR483-5p levels were increased in newly diagnosed pediatric ALL patients than ALL patients whose in remission (p<0.05).
miRNAs are thought to be identified at a different level of expression in normal and pathological tissues can be determined between the miRNAs that are effective diagnosis and treatment of human cancers. We showed the microRNA profils that may play new roles treatment of acute leukemia in the futures.

Disclosures:
Kilinc: ApoPharma Inc.: Research Funding.