bluuebunny/crossref_metadata_2025_split huggingface dataset:
Studies on Heme Oxygenase 1 During Erythroid Differentiation
Authors: Santos Daniel Garcia , Eisenberg Jesse , Schranzhofer Matthias , Ponka Prem 
Published: October 2019

Abstract
Abstract 4254
Heme is indispensable for the function of all aerobic cells as a prosthetic group of innumerable proteins. However, “free heme” (uncommitted) can initiate the formation of free radicals and cause lipid peroxidation, which can lead to cellular damage and tissue injury. Therefore, the rate of heme biosynthesis and catabolism must be well balanced by tight control mechanisms. The highest amounts of organismal heme (75-80%) are present in circulating red blood cells (RBC), whose precursors synthesize heme with rates that are at least one order of magnitude higher (on the per cell basis) than those in the liver – the second most active heme producer in the body. The degradation of heme is exclusively carried out by heme oxygenases 1 and 2 (HO1 and HO2), which catalyze the rate-limiting step in the oxidative degradation of heme. Although the heme-inducible HO isoform, HO1, has been extensively studied in hepatocytes and many other non-erythroid cells, virtually nothing is known about the expression of HO1 in developing RBC. Similarly, it is unknown whether HO1 plays any role in erythroid cell development under physiological or pathophysiological conditions. Using both a murine erythroleukemia cell line (MEL) and primary erythroid cells isolated from mouse fetal livers, we have demonstrated that during erythroid differentiation HO1 is up-regulated at both mRNA and protein levels. This increase in HO1 can be prevented by succinylacetone (SA), an inhibitor of heme synthesis that blocks 5-aminolevulinic acid dehydratase. These data suggest that in developing RBC, in addition to the continuous assembly of heme with globin chains, there is an increase in levels of uncommitted heme, which upregulates HO1 expression. Additionally, we have shown that down-regulation of HO1 via siRNA increased hemoglobinization in differentiating MEL cells. In contrast, induction of HO1 expression by NaAsO2 reduced the hemoglobinization of MEL cells. This effect could be reversed to control levels by the addition of HO1 inhibitor tin-protophorphyrin (SnPP). These results show that in differentiating erythroid cells the balance between levels of heme and HO1 have to be tightly regulated to maintain hemoglobinization at appropriate levels. Our results lead us to propose that disturbances in HO1 expression could play a role in some pathophysiological conditions such as thalassemias.

Disclosures:
No relevant conflicts of interest to declare.
Analysis of Rare deletional Thalassemia Using Custom CGH Array DNA Chip
Authors: Pissard Serge , Ceccaldi Raphael , Faubert-Laugé Emmanuelle , Velayoudane Valerie , Delattre Valerie , Lecerf Laure 
Published: October 2019

Abstract
Abstract 4278
Deletions are the main etiology for alpha thalassemia and account for 9 to 10 % of beta thalassemia. They are currently detected using “in house” methods as QMPSF or the use of commercially available assays as MLPA. The accurate determination of the breakpoint could be of importance for news and rare events to have a proper characterization of the deletion and to understand the consequences of the event. However, the imprecision due to the relative small numbers of markers in these methods, the distance to be covered and the high percentage of repeated sequences located into the intergenic regions, makes the breakpoint determination difficult. To overcome the limitation of the number of markers we have setup a custom “Comparative Genomic Hybridization” assay to study the beta and the alpha globin cluster. Based upon the Agilent multiple arrays system, we have designed an array covering chromosome 11 and chromosome 16, containing 15 000 probes with an average spacing of 150 bp on both clusters. This CGH DNA chip was used to successfully clone the breakpoint of several rare and new deletions removing from 10 kb to up to 1 megabase of DNA sequence on chromosome 11 and chromosome 16. The results show the efficiency of such an approach and the multiple array format allows a good cost value by comparison to step by step cloning. In 50 % from more than 100 beta locus deletional events the 5′ breakpoint lies between the Aγ globin and the delta globin gene indicating the location of hotspots of recombination herein. Recent findings have shown that intergenic sequences in β-globin cluster and probably in α-globin cluster may be involved in the global regulation of the globin genes expression. Interestingly, a non coding small RNA was recently discovered in the same region and supposed to be involved in fetal hemoglobin expression. Such multiple arrays CGH chips provide an easy and efficient tool to define critical intergenic regions involved in the global regulation of the globin genes. Such hypothesis is currently under study.

Disclosures:
No relevant conflicts of interest to declare.
RASGRP1/APTX Ratio Is a Strong Biomarker of Clinical Response and Survival In AML Patients Treated with Tipifarnib: A Phase I-II Preliminary Results
Authors: Martinelli Giovanni , Ottaviani Emanuela , Paolini Stefania , Papayannidis Cristina , Curti Antonio , Parisi Sarah , Iacobucci Ilaria , Cilloni Daniela , Arruga Francesca , Saglio Giuseppe , Russo Domenico , Visani Giuseppe , Zaccaria Alfonso , Rondoni Michela , Bocchia Monica , Candoni Anna , Fanin Renato , Gherlinzoni Filippo , Baccarani Michele 
Published: October 2019

Abstract
Abstract 4359
We conducted a phase I-II study aiming to assess efficacy and toxicity of tipifarnib-bortezomib treatment in elderly AML patients. RASGRP1/APTX genetic ratio, which is associated with treatment response in patients treated with tipifarnib alone, was tested.
Eighty patients were enrolled with secondary-AML: 14 had high risk cytogenetics; 42 were previously untreated. Seventy-five patients were treated.
Nine patients achieved complete remission (CR), 1 patient obtained a partial response (PR) and in 2 cases an hematological improvement (HI) was documented for an overall response rate (ORR) of 19%. Median time to response was 112 days, corresponding to 4 cycles (range 2–14). Marrow response (CR+PR) was significantly associated with overall survival (OS) (p<0.0001). RASGRP1/APTX was evaluated before treatment initiation on bone marrow (BM), peripheral blood (PB) or both. The median RASGRP/APTX value on BM was 15.3 (15-19.8) in responder patients and 2.2 (0.5-25.9) in non responders, respectively (p=0.00006). Its median value on PB was 31.6 (19.3-35.5) in responders and 6.4 (0.5-27.1) in non responders, respectively (p=0.00001). Interestingly, no marrow responses were recorded in patients with marrow RASGRP1/APTX ratio <8, while the response rate was 43% [how many were CR?] in patients with RASGRP1/APTX >8 (p<0.0001). Finally, RASGRP1/APTX levels significantly correlated with OS (p=0.001) with a median OS of 490 days and 162 days in patients with RASGRP1/APTX >8 and <8 respectively.
We conclude that the clinical efficacy of the combination of tipifarnib and bortezomib was evident. We confirmed that the RASGPR1/APTX BM or PB level is an effective predictor of response and survival and our now studying the response of such patients to tipifarnib, alone.
Supported by: European LeukemiaNet, AIL, AIRC, Fondazione Del Monte di Bologna e Ravenna, FIRB 2006, PRIN 2008, Ateneo RFO grants, Project of integrated program (PIO), Programma di Ricerca Regione – Università 2007 – 2009.

Disclosures:
Martinelli: Novartis: Consultancy, Honoraria; BMS: Consultancy, Honoraria; Pfizer: Consultancy. Baccarani:Novartis: Consultancy, Honoraria; BMS: Consultancy, Honoraria.
Primary Clinical Research of FA Regimen as Consolidation Therapy for Patients with Acute Myeloid Leukemia
Authors: Lu Huina , Qin Wei , Liang Aibin 
Published: October 2019

Abstract
Abstract 4380
In our study, 28 patients with AML treated in our department between 2004 and 2009 under complete remission, median age of 54 years (range 23–77) were consolidated with FA regimen which included fludarabine 30mg/m2 (days 1–3) and Ara-C 2g/m2 (days 1–3). Ara-C were administered as a continuous sequential infusion 4 hours later when the application of fludarabine had been over. Another 14 AML patients with matched conditions treated with high dose Ara-C (HDAra-C, 3g/m2, q12h×3d) were designated as controls. Event-free survival of both 2 groups of these patients were analysed.
Overall, 14 patients (50%) achieved continuous complete remission for more than 2 years. There is no statistically significant differences between FA group and HDAra-C group (P=0.214>0.05) in CCR. Patients aged >60 years also had no statistically significant differences compared to the patients aged <60 years in CCR. Seven patients relapsed after this treatment in a median survival of 11 months. Two of the relapsed patients had karyotype abnormalities and all of them had a markedly elevated WBC count at diagnosis.
The main toxicity was myelosuppression. WBC count and platelet count reached the lowest (about 0.8×10^9/L and 2.0×10^9/L) 6–13 days after the begining of the regimen. Median recovery time for granulocyte (≥1.0×10^9/L) and platelet (≥20×10^9/L) was 12 and 13 days, respectively. Fever(>38°C) caused by granulopenia occurred in 21 patients, recovered to normal for a median time of 5 days (range, 2–23). Heart, liver and enteric toxicity were negligible. There were no significant differences in hematologic toxicity or nonhematologic toxicity between the FA and HDAra-C group. The average hospitalization was about 20 days.
The FA is effective and safe as consolidation regimen in AML with acceptable toxicity and is not associated with an increased incidence of infections compared to conventional HDAra-C regimen.

Disclosures:
No relevant conflicts of interest to declare.
Homoharringtonine Affects the Expression of PI3K and p-AKT of NB4, SHI-1 Leukemia Cells
Authors: Yang Mingzhen , Zhang Xiaoyu , Huang Zhenqi , Liu Qinhua , Wang Lin , Li Qingsheng 
Published: October 2019

Abstract
Abstract 4381

Background:
Homoharringtonine (HHT) was efficient in therapying patients with acute myeloid leukemia (AML) in China, but little is known about the mechanism of its action. The phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling axis plays a central role in cell proliferation, growth, and survival under physiological conditions and many human cancers, including acute myelogenous leukemia (AML). We try to explore the effect of HHT on PI3K/Akt pathway in AML cells, thus supplying theoretical basis for wider use of HHT.

Method:
The NB4 and SHI-1 cells were cultured in 20% FCS RPMI-1640 with different concentration of HHT, cell proliferation was detected with MTT, apoptosis was measured by FCM, the protein of PI3K and p-Akt were determined by Western blot.

Result:
5ug/L HHT suppressed NB4 and SHI-1 cells proliferation and induced apoptosis after culture 24hr, 100ug/L HHT suppressed 71.29% NB4 and 64.83% SHI-1 cells proliferation respectively. Apoptosis increased obviously with the increasing HHT concentration and the culture time, the leukemia cell apoptosis was significant at 500ug/L HHT, about 41.84% NB4 cells and 46.88% SHI-1 cells were apoptosis when the HHT concentration was 100ug/L. The protein expression of PI3K, and p-Akt gradually declined with HHT concentration increasing, when 500ug/L HHT co-cultured with leukemia cells for 24 hours, The protein expression of PI3K and p-Akt were lowest. The p-Akt of NB4 and SHI-1 cells decreased 28.4% and 34.5% respectively at 5ug/L HHT for 48hr, the PI3K of NB4 and SHI-1 cells decreased 31.56% and 37.38% respectively at 10ug/L HHT for 48hr.

Conclusion:
HHT could inhibit NB4, SHI-1 cells proliferation and induce leukemia cells apoptosis, and could down-regulate the expression of PI3K and p-Akt significantly, this might be the one of mechanisms that HHT induce NB4 and SHI-1 cells apoptosis, we presume that HHT inhibit proliferation of acute myelogenous leukemia cells through effect of PI3K/Akt signaling pathways.

Disclosures:
No relevant conflicts of interest to declare.
Phenotyped Blood Reduces Alloimmunization Rates Among Cancer Patients
Authors: Novaretti Marcia Cristina Zago , Miyaji Sandra Cristina , Ito Gláucia Munemasa , Dinardo Carla Luana , Chamone Dalton FA 
Published: October 2019

Abstract
Abstract 4405

Background:
Alloimmunization against RBC antigens is a complication of blood transfusion/pregnancy that may cause hemolytic transfusion reactions and can difficult compatible blood availability in future transfusions. Frequency of transfusion, as well as patient's age, type of illness and treatment may influence the development of alloantibodies. Alloimmunization rates of 3.4% in hospitalized patients have been reported. The main purpose of this study is to compare the frequency of alloimmunization to red blood cell antigens in cancer patients that were transfused with phenotyped blood with those who were not.

Method:
Transfusion medical records of 669 oncologic patients were analyzed. Patients were evaluated by cancer type, number of transfusions, presence of alloantibodies and its clinical significance. Antibody screening and identification to red cell antigens were performed using gel-test (Diamed S/A, Lagoa Santa Brazil). All hematological patients were phenotyped for Rh (C, c, E, e, Cw) and K1 antigens using gel-test (Diamed S/A). Patients with hematological diseases were exclusively transfused with RBC units also matched for Rh/K1 antigens. Patients with other types of cancer were transfused with non-phenotyped RBC units. Patients that were previously immunized were excluded from the study.

Result:
Of 2,781 transfusions in 669 oncology patients, 1,026 were performed using phenotyped matched RBC units for Rh and K1 antigens. Patients with hematological diseases were more transfused when compared to patients with other types of cancer (p<0.001). However, alloimmunization rate found in patients that were transfused with phenotyped RBC units was statistically significant lower (1.026/1,000 transfusions) when compared to those one non-phenotyped blood (7.98/1,000 transfusions), (p=0.015). Moreover, the relative risk of a cancer patient to develop RBC alloantibodies when transfused with non-phenotyped Rh and Kell RBC units is 2.519 (IC 95%=2.181-2.908).
Conclusion: The rate of alloimmunization was statistically higher in patients that were transfused non-phenotyped RBC, even though this group was less transfused. We conclude that phenotyping RBC for Rh and K1 antigens reduces alloimmunization among cancer patients and prevents future complications when transfusing them.

Disclosures:
No relevant conflicts of interest to declare.
Procoagulant Activity of Laminaria Japonica Derived Fucoidan (BAX513) Depends on the Interaction with the C-Terminus of Tissue Factor Pathway Inhibitor
Authors: Palige Michael , Redl Christoph , Knappe Sabine , Ehrlich Hartmut J. , Dockal Michael , Scheiflinger Friedrich 
Published: October 2019

Abstract
Abstract 4417
BAX513, a fucoidan derived from the brown seaweed Laminaria japonica, and other non-anticoagulant sulfated polysaccharides (NASPs) improve coagulation in hemophilic blood and plasma. Fucoidans are heterogeneous, polysulfated molecules with procoagulant activities in a wide concentration range. Tissue factor pathway inhibitor (TFPI) has been described as a potential target for the procoagulant activity of NASPs (Liu et al. Thromb Haemost 2006; 95:68). In the current study, we investigated the interaction of BAX513 with TFPI proteins to gain a detailed understanding of the mechanism of action of BAX513. We used calibrated automated thrombography to monitor the activity of BAX513 in normal, FX and TFPI-deficient plasma. TFPI plasma levels were varied by the addition of truncated TFPI (TFPI1-160) and TFPI-domain specific antibodies. Initiating thrombin generation by addition of FXa to plasma deficient in both, FX and FVIII-showed a BAX513-dose dependent increase of thrombin generation, which was completely abolished when TFPI-specific polyclonal antibodies were present. Furthermore, when full-length TFPI was inhibited in plasma and instead supplemented with increasing amounts of TFPI 1–160, BAX513 did not show any activity. The data are further supported by surface plasmon resonance experiments (BiaCore) exploring the BAX513-TFPI interaction. A high affinity interaction was only observed for BAX513 with full-length TFPI but not for BAX513 with TFPI1-160. Our findings support a mechanism of action in which BAX513 acts as a potent dose-dependent TFPI antagonist that requires the highly charged C-terminus of TFPI to unfold its full potential. Understanding the mechanism of action of BAX513 supports the development of BAX513 as a promising new therapeutic for hemophiliacs and FVIII-inhibitor patients.

Disclosures:
Palige: Baxter Innovations GmbH: Employment. Redl:Baxter Innovations GmbH: Employment. Knappe:Baxter Innovations GmbH: Employment. Ehrlich:Baxter Innovations GmbH: Employment. Dockal:Baxter Innovations GmbH: Employment. Scheiflinger:Baxter Innovations GmbH: Employment.
All-Trans Retinoic Acid Modulates the Balance of ADAMTS13 and VWF In Human Microvascular Endothelial Cells
Authors: Wang Anyou , Yu Ziqiang , Dong Ningzheng , Ma Zhenni , Zhang Jingyu , Ruan Changgeng 
Published: October 2019

Abstract
Abstract 4424

Introduction:
All-trans retinoic acid (ATRA) can modulate the hypercoagulable state of human microvascular endothelial cell, which has been confirmed by the reports that ATRA can chang the expression of some procoagulant or anticoagulant factor, such as upregulation tissue factor and downregulation thrombomodulin. While, the balance between ADAMTS13 and von Willebrand factor (VWF) of microvascular endothelial cell, very related with the formation of microvascular platelet thrombsis, play an important role on the controlling the hypercoagulable state of microvascular endothelial cell. And it has been reported that the balance interrupted by some inflammatory cytokines may contribute to microvascular platelet thrombsis. In this study, to better understand the antithrombotic property of ATRA, we investigated whether ATRA may affect the balance.

Materials and Method:
We observed the effects of ATRA (compared to TNF-α) on the expression of ADAMTS13mRNA in human microvascular endothelial cell (HMEC-1 cell line) by real-time polymerase chain reaction amplification (RT-PCR). The levels of ADAMTS13 and VWF antigen in the conditioned medium of HMEC-1 cells were also detected by western blot or enzyme-linked immunosorbent assay (ELISA), and the proteolytic activity of ADAMTS13 was also determined by using GST-VWF73-His peptide as a specific substrate.

Result:
In a dose-dependent manner, ATRA significantly upregulated the expression of ADAMTS13mRNA which could be almost up to 4.63 times with 10uM ATRA, while TNF-α inhibited ADAMTS13mRNA expression which could drop to 41% of the control group with 10ng/ml TNF-α. ATRA also reverse the inhibition of TNF-α to ADAMTS13mRNA expression. The results were confirmed from the levels of ADAMTS13 protein and its activity, while ATRA had no significant affection on triggering release of VWF known as a specific substrate of ADAMTS13.

Conclusion:
This study provides the evidence that ATRA modulates the balance of ADAMTS13 and VWF in human microvascular endothelial cell, which may be a very relevant compartment for the antithrombotic property of ATRA.

Disclosures:
No relevant conflicts of interest to declare.
Preliminary Study of the Relationship Between EPO Receptor and Autoantibody on the Membrane of Erythropoietic Cells of the Patients with BMMNC- Coomb's Test(+) Hemocytopenia
Authors: Fu Rong , Liu Hui , Shao Zonghong , Li Lijuan 
Published: October 2019

Abstract
Abstract 4436

Objective
To observe the relationship between EPO receptor (EPOR) and autoantibody(IgG) on the membrane of erythropoietic cells of the patients with bone marrow mononuclear cell Coomb's(BMMNC-Coomb's) test(+) immuno-related pancytopenia(IRP),and then explore the probable autoantigens of autoantibodies in IRP.

Methods
42 newly diagnosed IRP patients (14 with IgG autoantibody on erythropoietic cells and 28 without this autoantibody) and 11 healthy donors as controls were enrolled in this study. EPOR on their nuclear erythrocyte were tested with flow cytometry. Their clinical data were also analyzed.

Results
(1) The level of EPOR of IgG(+) arm(1.59±0.87)% was significantly lower than that of IgG(-) arm (4.58±4.09)%(P<0.01), and the level of EPOR of IgG(-) arm was significantly higher than that of normal controls(2.27±1.76)%(P<0.05) G The level of EPOR of IRP patients was negatively correlated with their IgG autoantibody(r=-0.543,P=0.000) and its regression equation was Y(EPOR)=0.040-0.335X(IgG); (2) Reticulocyte percentage of IgG(+) arm (1.55±0.64)% was significantly lower than that of IgG(-) arm (2.41±1.42)%(P<0.05). The level of EPOR of IRP patients was positively correlated with reticulocyte percentage(r=0.346,P=0.029); (3)The serum level of indirect bilirubin(IDBIL) of IgG(+) arm was normal. While there were six patients (21.4%) whose serum level of IDBIL were higher than those of normal controls in IgG(-) arm.

Conclusion
The autoantibody(IgG) of some IRP patients might block or competitively inhibit the EPOR on the membrane of erythropoietic cells. It could inhibit the proliferation and differentiation of erythropoiesis in early phase.

Disclosures:
No relevant conflicts of interest to declare.