bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Characterization of Microparticle Phenotypes in Stored Red Blood Cell Units.
Authors: Tran Edward D , Wu Xiaoping , Blacken Grady R. , Lopez Jose A. , Dong Jing-fei , Fu Xiaoyun , Watkins Timothy R. 
Published: October 2019

Abstract
Abstract 2282
The transfusion of red blood cells (RBC) is associated with adverse outcomes in critically ill patients. One hypothesis explaining this observation involves the accumulation of potentially inflammatory/prothrombotic microparticles (MPs), released from cellular membranes during unit processing and storage. Despite their implication, the exact nature of MPs within RBC units remains incompletely defined. Here, we characterized the MP phenotypes present within stored RBC units and quantified the various populations. We directly sampled 11 O-positive non-leukoreduced RBC unit bags and isolated MPs by sequential centrifugation. We used a two-step process to detect MPs by flow cytometry. First, we used commercially available Megamix beads to define MPs according to their size during flow cytometric analysis and Trucount beads to quantify MP counts/mL (reported as median [IQR]). MPs ≤1 μm in size were subsequently classified into the following populations: total annexin V-positive, tissue factor-positive (CD142), and those staining for RBC- (intra/extracellular domain-specific glycophorin A, and band-3), leukocyte- (CD45), platelet- (CD42b), and endothelial-specific (CD144) markers. The total concentration of MPs was 1.52×109 MPs/mL [1.39×109]; however, only 4.3% were Annexin V-positive (5.16×107 MPs/mL [7.27×107]), and 11.7% were tissue factor-positive (4.24×107 MPs/mL [1.11×108]). Likewise, MPs with leukocyte-, platelet-, and endothelial-specific markers accounted for only 0.05% (4.44×105 MPs/mL [7.83×105]), 0.17% (1.99×106 MPs/mL [3.11×106]), and 0.06% (7.58×105 MPs/mL [8.04×105]), respectively (Figure 1). The amount of RBC-derived MPs varied depending on the antibody used. While only 4.2% of MPs (3.40×107 MPs/mL [2.91×107]) stained positive for the extracellular domain of glycophorin A, 41.3% (8.20×108 MPs/mL [4.06×108]) and 20.6% MPs (3.03×108 MPs/mL [3.53×108]) stained positive for the cytoplasmic domain of glycophorin A and band-3 respectively (Figure 1). The concentration of MPs positive for the cytoplasmic domain of glycophorin A was significantly greater their extracellular domain-specific counterparts (p<0.01). The results indicate that MPs in stored RBC units are primarily derived from RBCs, but also from other blood cells and vascular endothelial cells. A strong stain for the cytoplasmic domains of two RBC markers (cytoplasmic domain-specific glycophorin A and band 3) suggest the presence of a unique inside-out MP phenotype, potentially derived by RBC membrane “flipping” during microvesiculation. Ongoing studies are required to confirm these results and focus on 1) the mechanism of MP formation and (patho)physiological significance of inside-out RBC MPs and 2), the association between these MPs and the adverse effects seen in patients receiving RBC transfusions.

Disclosures:
No relevant conflicts of interest to declare.
Idarubicin Appears Equivalent to Dose-Intense daunorubicin for AML Induction
Authors: Trifilio Steven , Zhou Zheng , Mehta Jayesh , Czerniak Colleen , Pi Judy , Koslosky Molly , Greenberg Deborah , Pantiru Mihaela , Altman Jessica 
Published: October 2019

Abstract
Abstract 3581
Worldwide cancer drug shortages have recently required the substitution of daunorubicin, the “gold standard” anthracycline for idarubicin for induction therapy in acute myeloid leukemia(AML). Previously published randomized trials have shown idarubicin (12/m2) to be equal to, or better than daunorubicin (45–50mg/m2) for achieving complete remission(Blood. 1991 Apr 15;77(8):1666–74). It is currently unknown whether these results can be extrapolated when compared to intensified doses of daunorubicin(90mg/m2), shown to improve the CR rate when compared to 45 mg/m2 (N Engl J Med. 2009 Sep 24;361(13):1249–59). We therefore conducted an observational study to compare CR rates in AML patients who received either daunorubicin 90mg/m2 or idarubicin 12mg/m2 for the treatment of newly diagnosed AML.

Methods:
Medical records were used to identify newly diagnosed patients with AML who received daunorubicin 90mg/m2(Dau-90) or idarubicin 12mg/m2(Ida-12) between 1/2007 and 6/2012. Patients with APL were excluded. MDS patients who evolved to AML were included. AML was diagnosed by WHO criteria. Patients received daunorubin 90mg/m2 IV bolus or idarubicin 12mg/m2 IV bolus on days 1–3. Cytarabine 100mg/m2 by continuous infusion was given concomitantly on day 1–7 to both groups. All patients underwent marrow biopsy on day 14 after initiation of therapy. If residual disease was found, patients were considered treatment failure. Patients without residual disease were re-biopsied around the time of WBC recovery to assess CR. CR rates are reported after a single induction course.

Results:
28 Ida-12 and 37 Dau-90 patients were identified. Table 1 shows patient demographics and outcomes. There was no significant baseline difference between the 2 study arms for gender, age, weight, maximum WBC prior to therapy, cytogenetic risk group, percent of patients with FLT-3 ITD or NPM mutations, or combined risk factors. A non-significant trend was observed in the percent of marrow blasts prior to therapy and in the number of patients>60years(but not for age >55) within the Ida-12 study arm. Overall CR was similar between the Ida-12 and Dau- 90 treatment groups(p<0.1072) Subset analysis shows CR rates were significantly higher in Ida-12 treated patients who had an unfavorable karyotype(p<0.0359) or unfavorable combined cytogenetic risk(0.0388) or for age > 55 years(p<.0063). Overall morbidity and mortality was similar between groups.

Conclusion:
Ida-12 appears to be as effective as Dau-90 for achieving CR after a single induction course. Thus, Thus, substitution of Ida-12 for Dau-90 during times of drug shortages appears acceptable. Ida-12 may potentially be superior for patients >55 years or for those with unfavorable cytogenetics. These results warrant validation from a randomized clinical trial.

Disclosures:
No relevant conflicts of interest to declare.
Growth Inhibition of a Myeloid Leukemia Cell Line with 5q Deletion by Beta-Catenin Inhibition
Authors: Seba Amber D. , Wang Qi , Ming Ming , Qian Zhijian 
Published: October 2019

Abstract
Abstract 3608
As the population ages, the incidence of AML with complex cytogenetics continues to increase. These patients have a poor prognosis with 5% percent of patients diagnosed surviving 5 years. New strategies are needed to improve survival in AML patients. 5q deletion is present in 6–10% of cases of AML, and is a common chromosome abnormality in therapy-related AML (t-AML). 5q deletion is associated with haplo-insufficiency of the APC gene in hematopoietic stem cells and progenitors, leading to deregulation of the Wnt pathway and stabilization of beta- catenin (Wang, J Blood 2010). Loss of APC causes rapid exhaustion of hematopoietic stem cells and progenitors in vivo and leads to the accumulation of beta-catenin (Qian, Z et al JEM 2008). Beta-catenin is required for the development of leukemia stem cells in AML, but it is not required for the self-renewal of normal hematopoietic stem cells (Wang, Y et al Science 2010). Development of treatment strategies aimed at blocking the Wnt pathway through suppression of beta-catenin may improve and prolong remission rates.
The UoCM1 cell line maintains the unique characteristics of del(5q) leukemia (Qian et al, PNAS 2002). Western Blot shows that the expression of beta-catenin was higher in UoCM1 cells versus REH and MV4–11 cell lines, which do not have del(5q). To block the activation of Wnt signaling, through suppression of beta-catenin, we treated UoCM1, REH and MV4–11 cells with 100uM indomethacin. The UoCM1 cells showed 30% growth inhibition while REH and MV-4 cells displayed 10% growth inhibition. The down-regulation of beta-catenin in UocM1 is confirmed by Western Blot. Curcumin is a dietary polyphenol existing in Indian spice turmeric. We showed suppression of beta-catenin by curcumin in 293T cells by luciferase assay. Next, we treated UoCM1 and MV4–11 cells with 5uM or 10 uM curcumin. The growth of UoCM1 cells was significantly inhibited by curcumin as compared to MV4–11 cells. Treatment of UoCM1 cells with 10 uM curcumin produced 87% growth inhibition, while the growth inhibition was 29% for MV4–11 cells. 5uM concentrations of curcumin caused 48% growth inhibition in UoCM1 cells versus 16% in the MV4–11 cell line. Western Blot was performed to confirm that beta-catenin is down-regulated by curcumin, in the UoCM1 cell line.
In summary, blockade of the Wnt pathway through inhibition of beta-catenin suppresses cell growth in UoCM1 leukemia cells, with 5q deletion. Suppression of beta-catenin in combination with cytotoxic therapy may provide a novel means of treating leukemia in patients with a 5q deletion that both improves remission rates and prevents relapse.

Disclosures:
No relevant conflicts of interest to declare.
Outpatient Treatment of Symptomatic Pulmonary Embolism: A Systematic Review and Meta-Analysis
Authors: Piran Siavash , Le Gal Gregoire , Wells Phil , Righini Marc , Carrier Marc 
Published: October 2019

Abstract
Abstract 361

Background:
Patients with deep vein thrombus (DVT) can safely be treated as outpatients using subcutaneous low molecular weight heparin (LMWH). However the role of outpatient treatment in patients diagnosed with a pulmonary embolism (PE) is controversial. This practice is not accepted in many centers and majority of patients are still admitted to the hospital for observation. Clinicians are reluctant to discharge patients due to insufficient data supporting the safety of outpatient management of PE.

Purpose:
To determine the safety of outpatient treatment of acute symptomatic PE and the outcomes associated with outpatient management of PE including venous thromboembolism (VTE) recurrence, major bleeding, fatal PE, fatal intracranial hemorrhage (ICH), and overall mortality.

Methods:
A systematic literature search strategy was conducted using MEDLINE, EMBASE, the Cochrane Register of Controlled Trials and all EBM Reviews of published studies up to June 2012. We included randomized controlled trials (RCTs) and prospective cohort studies. We calculated the pooled proportions for the different outcomes at 3-months of follow-up. Ninety-five percent confidence intervals (95% CI) were calculated for each rate using the averaged, inverse variance-weighted estimates from each study.

Results:
Thirteen studies met all the inclusion criteria (3 RCTs and 10 prospective cohort studies). A total of 2458 patients were included in the systematic review. Different risk stratification methods were used to identify low risk patients with PE such as the pulmonary embolism severity index (PESI), the Geneva prognostic score, clinical gestalt and/or laboratory markers. The risk of recurrent VTE in patients with PE managed as outpatients was 1.64% (95% CI 0.69 – 3.0%; I2: 70.7%) during the 3 months follow-up period. The rate of fatal PE was 0.70% (95% CI 0.14 – 1. 7%; I2: 72.6%). The rates of major bleeding and ICH were 1.14% (95% CI 0.35 – 2.34%; I2: 72.8%) and 0.29% (95% CI 0.06– 0.68%; I2: 0%), respectively. The overall mortality rate was 2.3% (95% CI 0.41 – 5.7%; I-square: 92.8%). The I2 was moderate to high for most proportions, likely due to difference in the baseline characteristics (e.g. the inclusion of cancer patients in some studies).

Conclusion:
Independent of the risk stratification methods used, the rate of adverse events associated with outpatient PE treatment seems low. Based on our systematic review and pooled meta-analysis, numerous patients with acute PE can safely be treated as outpatients.
Proportion meta-analysis showing the risk of recurrent VTE in patients with PE managed as outpatients for a 3 month follow up period.

Disclosures:
No relevant conflicts of interest to declare.
High Efficiency of ICE (Ifosfamide-Carboplatin-Etoposide) in Relapse/Refractory Primary Central-Nervous System and Intra-Ocular Non Hodgkin Lymphoma, After First Line Treatment Containing High Doses of Methotrexate and Cytarabine. A Monocentric Retrospective Study From 2010 to 2012 On 17 Cases
Authors: Choquet Sylvain , roos Weil Damien , Xuan Khe Hoang , Cassoux Nathalie , Merle-Beral Helene , Leblond Veronique 
Published: October 2019

Abstract
Abstract 3664

Background:
Primary central nervous system lymphoma (PCNSL) and primary intra-ocular lymphoma (PIOL) are at very high risk of relapse after a first line treatment, and then carry a very poor prognosis. Autologous stem cell transplantation (ASCT) can offer prolonged responses but its results clearly depend on efficiency of salvage chemotherapy (Soussain, haemtologica, 2012). Since recent publications on first line treatment of PCNSL and PIOL recommend high dose methotrexate (Mtx) and cytarabine (AraC) (Ferreri, Lancet 2009), salvage chemotherapy must use other drugs with high level of penetration in the central nervous system (CNS). In this setting, ICE regimen, validated in systemic non Hodgkin lymphoma, seems to be appropriated but no data is published in PCNSL and PIOL.

Methods:
From june 2010 to may 2012, all relapse/refractory PCNSL and PIOL treated in first line by high doses of Mtx and AraC in the Pitie-Salpetriere Hospital, Paris, France, where treated by ICE regimen : ifosfamide (5g/m2 at day 2), carboplatine (AUC 5 at day 2) and etoposide (100mg/m2/d days 1 to 3). Doses where adapted on patient general status and ASCT proposed when possible.

Results:
Seventeen patients have been treated, 7 females and 10 males, median age 62 [28–84]. Four where refractory and 13 in relapse, with a mean progression free survival (PFS) of 368 days [85–1763], 4 had a second line, one a third before ICE. At moment of ICE treatment, localizations where 10 CNS, 2 CNS + PIOL, 3 PIOL and 2 meningitis. The mean number of cycles was 4 [1–6] and 4 patients needed a dose reduction. During treatment, grade 3/4 WHO toxicities where: 6 neutropenic fever (one death), 5 anemia, 9 neutropenia, 10 thrombopenia and one CNS complication (coma and hypersalivation). ASCT have been made in 6 patients (5 in CR, 1 in PR) and are pending in 3. Complete response (CR) have been obtained in 13 patients (76%), partial in 2. With a mean follow-up of 405 days, 6/15 patients in response relapsed (only one after ASCT), in a median of 81 days, 9 patients died (7 by progression, one during treatment and one in CR). Median Overall survival (OS) was 220 days for all patients but was not reached in case of ASCT.

Conclusion:
ICE regimen is very effective in relapse/refractory PCNSL and PIOL heavily treated by high dose Mtx and AraC. This efficacy can allow to perform ASCT in eligible patients, chemosensitivity being the most important factor influencing the OS and PFS after ASCT. ICE can represent a new standard in this setting.

Disclosures:
Leblond: Roche: Advisory Board Other, Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Mundipharma: Honoraria; Janssen-Cilag: Honoraria.
R-HCVAD Alternating with R-Methotrexate Cytarabine in Younger Patients (pts) with IH and High-Risk Age Adjusted-IPI DLBCL
Authors: Oki Yasuhiro , Westin Jason R. , Fowler Nathan , Neelapu Sattva , Hagemeister F.B. , McLaughlin Peter , Kwak Larry W. , Romaguera Jorge Enrique , Fanale Michelle , Younes Anas , Rodriguez Maria A. , Orlowski Robert Z. , Wang Michael , Ouzounian Souzanne T , Samaniego Felipe , Fayad Luis 
Published: October 2019

Abstract
Abstract 3707

Background:
The management of pts with untreated, high-risk DLBCL needs improvement. R-HCVAD/R-MTX-ARA-C is a regimen with significant activity in Burkitt's, ALL, and mantle cell lymphomas. The aim of this phase II study is to evaluate the efficacy and safety of R-HCVAD in patients up to 60 years with IH or High risk DLBCL by age adjusted IPI.

Methods:
Pts with untreated indolent DLBCL with aa-IPI score of 2 or 3 risk were eligible. Treatment was R-HCVAD alternating with R-Methotrexate-cytarabine unless the patient had an effusion that precluded from receiving methotrexate. Prophylactic antibiotics and growth factors were routinely given to pts, and intrathecal prophylaxis were given in patients who were considered to be at high risk for CNS recurrence. Response was assessed every 2 cycles using the International Working Group Response Criteria 2007 incorporating PET scan.

Results:
A total of 48 pts received at least one cycle of treatment and were evaluable for toxicity. Forty-five were evluable for response. Pts characteristics are the followings; male 60%, median age 50 (24–60), elevated LDH 92%; stage IV 79%, III 19%, PS>1 69%, AA IPI IH 42%, High 58%. Pts received a median of 6 cycles (1–6). Two patients died after 1stcycle, 1 due to sepsis, another pulmonary embolism. Another patient refused treatment after the second cycle and died of disease. CR was observed in 83%,PR in 6%, PD 2%. At the moment of this evaluation, with a median follow of 30 months, 35 (73%) are alive in first remission, 2 (4%) died in CR of glioblastoma and anal cancer, 5 (10%) died of disease, 2 are alive with disease, 2 are alive after transplant, and 2 patients died of toxicity.

Conclusion:
R-HCVAD/R-MTX-Cytarabine is a very active regimen in DLBCL with high-risk features. We however observed significant hematological toxicities and early death in this study.

Disclosures:
Fanale: Genetech: Research Funding. Younes:Seattle Genetics: Research Funding; Allos: Research Funding. Wang:Onyx Pharmaceuticals: Honoraria, Research Funding. Fayad:Roche: Research Funding.
Development and Characterization of a Novel Human Xenograft Model Using an MDS Patient-Derived Cell Line
Authors: Rhyasen Garrett , Wunderlich Mark , Mulloy James C. , Starczynowski Daniel T 
Published: October 2019

Abstract
Abstract 3814
Myelodysplastic syndromes (MDS) are hematopoietic stem cell disorders, defined by ineffective hematopoiesis, blood cytopenias, myeloid dysplasia, and an increased risk of acute myeloid leukemia (AML). Immunodeficient mice engineered to support human tumor xenografts represent a pivotal development in the study of human cancers. The xenograft model has been an attractive approach to interrogate human AML; in contrast, MDS patient samples exhibit negligible or poor engraftment, with no evidence of disease in immunocompromised recipient mice. To circumvent this limitation we developed a novel xenograft mouse model using an MDS cell line, MDSL. The MDSL cell line was derived from the non-leukemic phase of an MDS patient with refractory anemia-ringer sideroblasts, and has maintained growth-factor dependence and sensitivity to Lenalidomide. Two immunocomprismised mouse stains were used for xenotransplantation of MDSL cell: NOD/LtSz-scid IL2RG (NSG) and NOD/LtSz-scid IL2RG-SGM3 (NSGS). NSGS mice are identical to NSG except that they have been engineered to express three human cytokines, SCF, GM-CSF and IL-3, to improve engraftment of human hematopoietic cells. Following transplantation of MDSL cells by either intrafemoral or intravenous injection, NSG and NSGS animals exhibited human cell grafts in the bone marrow, peripheral blood, and spleen. Subsequently, mice developed a fatal disease exhibiting features of human MDS, including anemia and thrombocytopenia, which coincides with clonal expansion and increased human cell graft in all hematopoietic organs. The median survival for NSGS and NSG mice was 21 and 55 days post-injection, respectively. The shortened latency in NSGS mice suggests that the expression of human cytokines accelerates the growth and enhances survival of the MDSL cells in vivo. MDSL cells isolated from the bone marrow of primary recipient mice were capable of initiating an MDS-like disease in secondary recipients. Given that few preclinical options for MDS are available, we also evaluated whether this model is suitable for therapeutic studies. Since MDSL cells are sensitive to Lenalidomide and IRAK1 Inhibitor (Rhyasen et al, ASH 2011), we evaluated the effects of these drugs in the xenograft model. After xenotransplantation of MDSL cells, NSGS mice were administered either Lenalidomide or IRAK1 Inhibitor. Our preliminary findings indicate that mice xenografted with MDSL cells are amenable to drug delivery and show a response to treatment. In summary this work describes a novel xenograft model of human MDS, the first to develop clinical features resembling human disease. In addition, we predict that this model will also be useful for preclinical studies to examine novel and existing MDS therapies.

Disclosures:
No relevant conflicts of interest to declare.
Metabolites of Tryptophan Catabolism Are Elevated in Sera of Patients with Myelodysplastic Syndromes and Inhibit Hematopoietic Progenitor Amplification
Authors: Berthon Céline , Fontenay Michaela , Corm Selim , Briche Isabelle , Lhermitte Michel , Quesnel Bruno 
Published: October 2019

Abstract
Abstract 3843

Introduction:
Tryptophan catabolism, which is mediated by the enzymes indoleamine 2,3-dioxygenase (IDO) and tryptophan 2,3-dioxygenase (TDO), produces kynurenine, which blocks T-cell activation and induces immunosuppression. Kynurenine itself is converted by downstream enzymes into secondary catabolites that also have toxic effects on T cells. Tryptophan catabolism is elevated in many cancers, including acute myeloid leukemia (AML). However, tryptophan catabolites that are downstream of kynurenine have never been investigated in hematological malignancies.

Methods:
We evaluated the serum levels of primary and secondary tryptophan catabolites in a cohort of patients with myelodysplastic syndromes (MDS). Sera were isolated from 132 adult MDS patients after informed consent was obtained in accordance with the Helsinki Declaration. The levels of tryptophan, kynurenine, kynurenic acid, 3-hydroxykynurenine, 3-hydroxyanthranilic acid, and anthranilic acid in the sera were quantified using HPLC. For erythroid cell expansion, CD34+ cells were collected and isolated from the mononuclear cell fractions of cytapheresis products from 3 healthy donors and cultured in liquid medium under erythroid conditions with tryptophan catabolites.

Results:
The MDS patients showed significantly lower levels of tryptophan and higher levels of kynurenine, kynurenic acid, 3-hydroxyanthranilic acid, and anthranilic acid compared with the healthy controls. We also compared the kynurenine and tryptophan levels in the MDS patients with our previous cohort of 112 patients with primary AML. The kynurenine/tryptophan ratios were significantly higher in the MDS patients (median 0.0468 vs. 0.0676). The tryptophan catabolites correlated with cytopenia; higher kynurenine levels were associated with lower hemoglobin levels and higher blast counts and were associated with presence of dysgranulopoiesis. Lower tryptophan levels were found in patients with platelet transfusion dependency. Kynurenic acid levels were higher in patients with dysmegakaryopoiesis. High 3-hydroxyanthranilic and kynurenic acid levels were associated with severe thrombopenia below 20 G/L. IPSS score, cytogenetic, and WHO diagnosis did not associated with any tryptophan catabolite level. The tryptophan catabolites inhibited progenitor expansion during the in vitro culture of hematopoietic cells and reduced the numbers of granulocytes and erythroblasts.

Conclusions:
Thus, MDS patients are characterized by high tryptophan catabolism resulting in elevated primary and secondary metabolites, which both have inhibitory effects on hematopoiesis. These results suggest that IDO or TDO inhibitors should be investigated in MDS.

Disclosures:
No relevant conflicts of interest to declare.