bluuebunny/crossref_metadata_2025_split huggingface dataset:
Reduced Packed Red Blood Cell Utilization Via Implementation of Transfusion Guidelines in a Large Health System Hospital
Authors: Bowers Reid Christian , Thompson Michael A. , Van Dreser Julie , Sanchez Federico Augusto 
Published: November 2019

Background: We have previously discussed how implementation of Patient Blood Management (PBM) principles such as transfusion guidelines helped reduce packed red blood cell (PRBC) usage in cardiovascular surgery patients (Bowers et al, Reduced Blood Product Utilization via Implementation of an Anemia Clinic and Consult Service in a Large Health System Hospital https://tinyurl.com/y3ucr3sz). We now discuss the effect of implementing transfusion guidelines on PRBC transfusion practices at an institutional level at St. Luke's Medical Center, Advocate Aurora Health in Milwaukee, WI.
Methods: Beginning in 2015 Aurora Health Care made 7 g/dL the standard threshold below which packed red blood cell (PRBC) transfusion is considered appropriate. Additionally, transfusing PRBCs as single units with a hemoglobin (Hgb) measurement between each unit was made standard. Education was provided to clinicians on the new standard including best practice advice in the transfusion ordering workflow. Data was gathered on institutional PRBC use and individual clinician PRBC ordering practices. Further education was provided based on the extent to which data showed conformity to the new standard.
Results: PBM implementation 2015-2018 was associated with a drop in institutional PRBC usage from 11,490 to 9301 units. The percentage of transfusions ordered with Hgb &lt; 7 g/dL rose from 41.6% to 71.8%. The percentage of single unit orders increased from 67.3% to 91.8%. See Graph/Table 1 & 2 for further analysis of individual practitioner PRBC ordering. See Table 3 for reporting period volume data.
Conclusions: Implementation of PRBC transfusion guidelines was associated with a greater than 2000-unit annual reduction in PRBC usage. Reduction was associated with individual clinician transfusion practices that increasingly conformed with guidelines over time.

Disclosures
Thompson: Adaptive: Membership on an entity's Board of Directors or advisory committees; AbbVie: Research Funding; UpToDate: Patents & Royalties: Myeloma reviewer; BMS: Research Funding; Celgene: Membership on an entity's Board of Directors or advisory committees; Lynx Bio: Research Funding; VIA Oncology: Membership on an entity's Board of Directors or advisory committees; GSK: Membership on an entity's Board of Directors or advisory committees; Doximity: Equity Ownership, Membership on an entity's Board of Directors or advisory committees; Takeda: Membership on an entity's Board of Directors or advisory committees, Research Funding.
Use of Multi-Omics Approach to Produce a Robust Serum-Free Media for the Expansion of T Cells
Authors: Mansour Sarya G. , Lin Pei-Yi , Pierce Anson , Civin Curt I 
Published: November 2019

Chimeric antigen receptor (CAR) T-cell therapy represents a major advancement in personalized cancer treatment. Generating robust and well characterized manufacturing processes for cell therapy has become crucial. Traditional manufacturing processes activate and expand T cells in media containing human serum, which supports cell growth and viability. However, serum batches vary significantly from lot to lot and require frequent screening for contaminants that can be detrimental to patients. T cell manufacturing processes independent of human serum will render adoptive T cell therapy less expensive, more consistent, and safer for patients. In order to create a best-in class serum-free medium that is even more robust than serum-containing media, we need to gain a better understanding of the metabolic requirements of T cells during expansion. For this purpose, we have utilized a multi-omics approach to fully characterize the proteomic and metabolomic signatures of T cells expanded in a serum free, xeno free medium at different phases of growth. We sampled cells at days 3, 5, and 7 for label-free proteomics and untargeted extra- and intracellular metabolomics analysis in order to identify metabolic patterns that could be corrected with media supplementation. We identified over 6,043 proteins and 900 metabolites from 4 donors and detected 1,200 significantly different proteins and 312 metabolites. Using an in-house bioinformatics strategy to analyze the multi-omics data, we focused on several metabolic and signaling pathways that were significantly different at days 5 and 7 compared to day 3. The "corrective" media supplementation that we added based on these results demonstrated a tremendous increase in cell growth and viability yielding a 2.6- and 3.2-fold increase in cell growth on days 5 and 7, respectively, and an increase in viability of 15-20% and 11-15% on days 5 and 7, respectively, compared to the un-supplemented prototype. In addition, cells maintained high viability throughout the whole culture and phenotype was not affected. These data demonstrate that multi-omics is a powerful tool for understanding T cell metabolism and identifying components to develop robust and reproducible serum-free media that produces high quality T cells.

Disclosures
No relevant conflicts of interest to declare.
The Diagnostic Performance of a Non-Emergency Department-Based Deep Vein Thrombosis Ambulatory Care Pathway
Authors: Sene Pape-Mamadou , Yahiaoui Denis , Koolian Maral , Tagalakis Vicky 
Published: November 2019

Background
Ambulatory management of isolated acute deep vein thrombosis (DVT) is the standard of care in most patient populations, and in clinical practice is mostly initiated in the emergency department (ED). However, patients referred to the ED with suspected DVT often experience unnecessary delays to diagnosis and subsequent long stays to determine appropriate treatment and follow up care. We implemented a DVT ambulatory care pathway that does not include the ED for referred patients with suspected isolated DVT that begins with pre-test risk stratification in the community and subsequent algorithm-based diagnostic work-up, treatment, and follow-up at a tertiary care centre in Montreal Canada.
Objective
To determine the diagnostic performance of a non-ED based DVT ambulatory care pathway.
Methods
We determined the prevalence of DVT over a 46 week period between August 2018 and June 2019 among ambulatory patients with suspected isolated DVT who following community-based risk stratification using the modified Well's clinical prediction score were referred for diagnostic work up and treatment using a newly implemented non-ED based DVT ambulatory care pathway.
Results
Among 122 patients referred by community physicians, 86 (70%) met pre-defined pathway criteria for assessment of suspected DVT. In all, 42 (49%) were referred with an unlikely/low modified Well's score and 44 (51%) with a likely/high score. Overall, the prevalence of DVT was 19.8%, specifically 9.5% in the unlikely/low and 30.2% in the likely/high pretest probability groups, respectively.
Conclusion
Our results show that the diagnostic performance of a non-ED based acute DVT ambulatory care pathway is in line with literature estimates. The advantage of this pathway is that it offers clear, evidence-based guidance for community physicians to diagnosis and treat patients in an ambulatory setting without using the ED. The approach is likely to result in both healthcare and economic benefits, including increased patient satisfaction and shorter ED stays.

Disclosures
Tagalakis: Sanofi Aventis: Other: investigator initiated grant;participated on ad boards; Pfizer: Other: participated on ad boards; BMS-Pfizer: Other: participated on ad boards; Servier: Other: participated on ad boards; Bayer: Other: participated on ad boards.
Anaerobic Antibiotics and the Risk of Graft-Versus-Host Disease after Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Tanaka John , Young Rebecca , Spees Lisa , Jenkins Kirsten , Sung Anthony D. , Martin Paul L. , Heston Sarah , Thompson Jillian C. , Seed Patrick , Arshad Mehreen , Kelly Matthew 
Published: November 2019

Background: The gut microbiota interacts extensively with the host immune system and thus may modify the risk of graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation (HSCT). During the post-transplant neutropenic period, the majority of allogeneic HSCT recipients receive empirical broad-spectrum antibiotics for febrile neutropenia. We hypothesized that receipt of an antibiotic regimen with an anaerobic spectrum of activity is associated with a higher risk of grade II-IV acute GVHD than receipt of a non-anaerobic antibiotic regimens.
Methods: In this single-center retrospective cohort study, we evaluated associations between peri-transplant receipt of antibiotics with an anaerobic spectrum of activity and the risk and severity of GVHD among 877 adults who received an allogeneic HSCT between January 1, 2005 and December 31, 2016. We identified 609 patients who developed febrile neutropenia after HSCT and compared GVHD risk and mortality among patients who received anaerobic antibiotics (piperacillin-tazobactam or carbapenems; n=333) to patients who received only antibiotics with minimal activity against anaerobes (aztreonam, ceftazidime, or cefepime; n=276). Antibiotics received by patients between 7 days before and 28 days after allogeneic HSCT and GVHD diagnoses were verified via manual review of medication orders and provider notes in electronic medical records.
Results: Receipt of anaerobic antibiotics was associated with an increased risks of grade II-IV acute GVHD (hazard ratio (HR): 1.41; 95% confidence interval (CI): 1.10-1.79; P=0.01) and acute GVHD mortality (HR: 1.87; 95% CI: 1.13, 3.11; P=0.02). This hazard was primarily associated with acute GVHD of the gut or liver (HR: 1.38; 95% CI: 1.06, 1.79; P=0.02). The association remained with even short (<7 days) courses of anaerobic antibiotics. Anaerobic antibiotic exposure was not associated with acute skin GVHD (HR: 0.97; 95% CI: 0.69, 1.37; P=0.88), chronic GVHD diagnosis (HR: 0.93; 95% CI: 0.70, 1.23; P=0.43), or chronic GVHD mortality (HR: 0.89; 95% CI: 0.44, 1.81; P=0.76).
Conclusions: Receipt of anaerobic antibiotics for febrile neutropenia post-HSCT is associated with an increased risks of acute GVHD of the gut or liver and acute GVHD mortality. Limiting use of antibiotics with an anaerobic spectrum of activity after allogeneic HSCT may reduce acute GVHD incidence and mortality.

Disclosures
Sung: Novartis: Research Funding; Merck: Research Funding; Seres: Research Funding. Martin:Novartis Pharmaceuticals: Other: research support; Jazz Pharmaceuticals: Other: research support.
Excess Serine from the Microenvironment Caused the Impaired Megakaryopoiesis and Thrombocytopia in Multiple Myeloma
Authors: Kuang Chunmei , Xia Meijuan , An Gang , Liu Cuicui , Wu Dan , Su Pei , Zhang Jingyu , Xia Jiliang , Guo Jiaojiao , Wu Xuan , Jia Wei , Qiu Lugui , Zhou Wen , Zhou Jiaxi 
Published: November 2019

Background: Thrombocytopenia is major complication in a subset of patients with multiple myeloma (MM). However, a lack of detailed studies about the megakaryopoiesis, thrombopoiesis as well as their connections with the survival of patients limits us to explore whether thrombocytopenia could be used as a reliable prognostic factor for MM.
Materials and Methods: In this study, 1393 newly diagnosed MM patients were selected for investigating the potential connection between PLT counts and clinical characteristics including ISS stage, overall survival as well as progression free survival. Besides, 5T33MMvt-KaLwRij mouse model were also used to examine the megakaryopoiesis and thrombopoiesis during disease progression. The proportion and function of different subpopulations of cells including megakaryocytes, megakaryocytic-erythroid progenitors (MEPs), common myeloid progenitors (CMPs) and Lin-Sca-1+c-kit+ (LSK) cells were measured both in MM patients and mouse models. Gas chromatography-time-of-flight mass spectrometry (GC-TOFMS)-based metabolomics was used to analyze the metabolites.
Results: Of the 1393 studied patients, 298 cases of MM patients are found with thrombocytopenia at the time of diagnosis. PLT counts were lower both in stage Ⅱ (P<0.01) and Ⅲ patients (P<0.001) than stageⅠpatients. Interesting, we found MM patients with thrombocytopenia had a significantly lower OS (P<0.001) and PFS (P<0.001). In mouse model, we also found PLT counts gradually decreased in peripheral blood during the disease progression (P<0.001). Further analysis demonstrated the proportion and the absolute numbers of megakaryocytes and MEPs were diminished both in mouse and MM patients with thrombocytopenia. PLT counts were negative correlated with the percentage of plasma cells or IgG2b levels (P<0.001), suggesting a potential connection of malignant cells infiltration and thrombocytopenia. In mechanism, metabolomics analysis with BM plasma identified 16 differential metabolites in MM patients with thrombopoiesis (VIP > 1.2, P < 0.05). Among them, serine was observed significantly be elevated in MM mouse and suffice to inhibit the megakaryopoieis and thrombopoiesis in vitro.
Conclusion: PLT counts might be used as a reliable prognostic factor for MM patients since the thrombocytopenia was associated with poor survival in MM patients. The thrombocytopenia in MM might attribute to, at least partially, the inhibition by serine from the microenvironment. Our findings revealed novel mechanism of MM and might eventually shed light on the treatment of MM patients.

Disclosures
No relevant conflicts of interest to declare.
Effects of Interferon-γ on Proliferation and Ability of Secretion of Human Amniotic Mesenchymal Stem Cells
Authors: Gao Ya , Xu Ying , Li Weiru , Zhang Yintian , Ping Baohong , Liu Qifa 
Published: November 2019

Objective:The immunoregulatory properties and proliferation of mesenchymal stem cells (MSCs) could be affected by inflammatory factors. However, there have been few studies about human amniotic MSCs (hAMSCs). We investigated the effects of interferon (IFN)-γ on the proliferation and apoptosis of hAMSCs, and measured the level of inflammatory factors secreted by hAMSCs. Result:hAMSCs were cultured with complete medium with different concentrations of IFN-γ. We detected the proliferation of hAMSCs by Cell Counting Kit-8 assays, analysed apoptosis by flow cytometry (FCM) at 48 h, and mesasured the level of inflammatory factors such as solube HLA-G and prostaglandin E2 (PGE2) in the supernatant at 48 h by ELISA. The level of kynurenine (KYN) was measured by ultraviolet spectrophotometry. As culture time increased, the proliferation of hAMSCs with different concentrations of IFN-γ increased rapidly from day 1 to day 4, and then the growth rate slowed. FCM indicated that there was no significant apoptosis in the 100 ng/ml IFN-γ group compared with cells without IFN-γ. The level of PGE2 and soluble HLA-G in cells with IFN-γ was higher compared with those without IFN-γ. The level of KYN increased significantly in the cells with IFN-γ. Conclusion:IFN-γ did not affect the growth and proliferation of hAMSCs, and promoted secretion of PGE2 and soluble HLA-G, and enhanced activity of indoleamine 2,3-dioxygenase (IDO), providing a theoretical basis for hAMSCs to prevent and treat graft-versus-host disease.

Disclosures
No relevant conflicts of interest to declare.
Anti-BTN3A 20.1 Agonist Monoclonal Antibody Enhances Autologous Vγ9Vδ2 T Cells Cytotoxicity Against Primary Acute Myeloid Blasts
Authors: Le Floch Anne-Charlotte , Orlanducci Florence , Briantais Antoine , Le Roy Aude , Chretien Anne-Sophie , Vey Norbert , Olive Daniel 
Published: November 2019

Introduction: Vγ9Vδ2 T cells immunotherapy is an emerging strategy in acute myeloid leukemia (AML). Partial effects of phosphoantigens like zoledronate, prompted us to investigate other molecules. Using Vγ9Vδ2 T cells expanded from healthy volunteers, recent works of our team demonstrate that anti-BTN3A 20.1 agonist monoclonal antibody (mAb) improves Vγ9Vδ2 T cells cytotoxicity in vitro and in vivo, including in zoledronate resistant blasts. Efficacy of anti-BTN3A 20.1 agonist mAb remains undetermined using autologous Vγ9Vδ2 T cells and mechanism of its action on AML blasts is understudied.
Material and methods: Expression of co-stimulatory molecules on surface of AML blasts and Vγ9Vδ2 T cells, from 37 PBMC of AML patients, was assessed by flow cytometry. Vγ9Vδ2 T cells expansion was realized from the same samples with zoledronate, and administrations of Il-2 and Il-15. Purity was assessed by flow cytometry at day 13. Degranulation assay was performed at day 14.
Results: 20 degranulation assays were realized on successfully expanded Vγ9Vδ2 T cells (69% of analyzable samples). Degranulation with anti-BTN3A 20.1 agonist mAb was enhanced compared to zoledronate condition (p=0.0085) or negative control (p<0.0001). We also show a positive correlation between level of degranulation and expression of DNAM-1 on Vγ9Vδ2 T cells surface, exclusively under anti-BTN3A 20.1 agonist mAb treatment (rS=0.54; p=0.0134). Conversely, under zoledronate treatment, Vγ9Vδ2 T cells degranulation was positively correlated with expression of NKG2D on Vγ9Vδ2 T cells surface (rS=0.62; p=0.0099) and was inversely associated to their expression of PD-1 (p=0.0482).
Conclusion: Efficacy of anti-BTN3A 20.1 agonist mAb on primary AML blasts is confirmed in an autologous setting. Degranulation seems to play a major role in enhancement of Vγ9Vδ2 T cells cytotoxicity under anti-BTN3A 20.1 agonist mAb treatment, where DNAM-1 could play a determinant role. An autologous strategy for Vγ9Vδ2 T cells may be studied and considered in AML patients, with potent extrapolation to γδ T cell therapy protocols in targeting strategies.

Disclosures
Vey: Novartis: Consultancy, Honoraria; Janssen: Honoraria. Olive:ImCheck Therapeutics: Consultancy, Equity Ownership, Patents & Royalties; GlaxoSmithKline: Patents & Royalties.
Calcineurin Inhibitors Replacement By Ruxolitinib As Graft Versus Host Disease Prophylaxis for Patients after Allogeneic Stem Cell Transplantation
Authors: Zhao Yanmin , Huang He 
Published: November 2019

Severe graft versus host disease (GVHD) is a leading cause of morbidity and mortality after allogeneic stem cell transplantation (allo-SCT). Currently, the most widely used prophylaxis regimen usually includes calcineurin inhibitor (CNI; i.e., tacrolimus and cyclosporine). However, the incidence of aGVHD is about 30%-50%, and a minority of patients have contraindications against CNI. Thus, improved GVHD prevention methods are still needed. This study aimed to determine the prophylactic value of ruxolitinib for graft versus host disease (GVHD) in calcineurin inhibitor (CNI)-intolerant patients after allogeneic stem cell transplantation (SCT). From September 2017 to March 2019, 10 patients intolerant to CNI were enrolled. The regimens were based on the myeloablative BuCy regimen. Thymoglobulin 6 mg/kg was applied to patients with HLA-haploidentical related donors (HRD). All received ruxolitinib to replace CNI as GVHD prophylaxis. Ruxolitinib was initiated at 5-10 mg twice daily, maintained for 2-3 months, and then tapered gradually. Eight patients had acute leukemia, one had myeloproliferative neoplasm, and one had natural killer T-cell (NK/T-cell) lymphoma. The donors were from matched sibling donors (MSD, n = 3) and HRD (n = 7). They received CNI plus short-time methotrexate as GVHD prophylaxis, but all showed intolerance within 45 days after SCT due to transplantation-associated thrombotic microangiopathy (n = 4), CNI-induced pain syndrome (n = 2), and CNI-related renal/hepatic dysfunction (n = 4).After ruxolitinib replacement, only one patient (10%) developed grade II skin aGVHD within 100 days, and only one developed severe aGVHD after 100 days. Among nine patients who survived beyond day +100, two (2/9, %) developed moderate/severe cGVHD after tapering or stopping ruxolitinib, resulting in 1-year accumulative incidence of moderate/severe cGVHD of 23.8%. Cytomegalovirus reactivation occurred in four patients (40%). After a median follow-up of 10.5 (range: 2-14.5) months, two out (20%) relapsed, and seven (70%) were alive in good condition, of whom six (60%) maintained negative MRD.This study showed that the prophylactic application of ruxolitinib for CNI-intolerant patients after allo-SCT was effective in preventing GVHD, with a mild side effect.

Disclosures
No relevant conflicts of interest to declare.
The Fanconi Anemia-Associated Protein NIPA Is Essential for the Nuclear Abundance of FANCD2
Authors: Kreutmair Stefanie , Erlacher Miriam , Andrieux Geoffroy , Istvanffy Rouzanna , Rudorf Alina , Zwick Melissa , Rückert Tamina , Pantic Milena , Poggio Teresa , Shoumariyeh Khalid , Müller Tony Andreas , Kawaguchi Hiroyuki , Follo Marie , Klingeberg Cathrin , Wlodarski Marcin , Baumann Irith , Pfeifer Dietmar , Kulinski Michal , Rudelius Martina , Lemeer Simone , Kuster Bernhard , Dierks Christine , Peschel Christian , Cabezas-Wallscheid Nina , Duque-Afonso Jesus , Zeiser Robert , Cleary Michael , Schindler Detlev , Schmitt-Graeff Annette , Boerries Melanie , Niemeyer Charlotte M. , Oostendorp Robert A.J. , Duyster Justus , Illert Anna Lena 
Published: November 2019

Inherited bone marrow failure syndromes (IBMFS) are a heterogeneous group of disorders characterized by impaired stem cell function resulting in pancytopenia. Diagnosis of IBMFS presents a major challenge due to limited diagnostic tests and overlapping phenotypes. For that reason, novel and clinical relevant biomarkers and possible targets are urgently needed.
Our study defines NIPA as an IBMFS gene, which is significantly downregulated in a distinct subset of MDS-type refractory cytopenia of childhood patients. Mechanistically, NIPA binds FANCD2 and regulates its nuclear abundance. The stabilization of both non- and monoubiquitinated FANCD2 identifies NIPA as an essential player in the Fanconi Anemia (FA) pathway. NIPA thereby prevents MMC hypersensitivity visualized by increased numbers of chromosome radials and reduced cell survival after induction of interstrand crosslinks. To provide proof of principle, re-expression of FANCD2 in Nipa deficient cells restores MMC sensitivity. In a knockout mouse model, Nipa deficiency leads to major cell intrinsic long-term repopulation defects of hematopoietic stem cells (HSCs), with impaired self-renewal in serial transplantations and a bias towards myeloid differentiation. Unresolved DNA damage in Nipa deficient HSCs causes increased sensitivity to cell death and leads to progressive, age-related loss of the HSC pool. Induction of replication stress triggers the phenotypic reduction and functional decline of murine HSCs, resulting in complete bone marrow failure and death of the mice thereby mimicing Fanconi Anemia.
Taken together, our study adds NIPA to the short list of FA-associated proteins being essential for a functional DNA repair/FA/BRCA axis and thereby emphasizing its impact as potential diagnostic marker and/or possible target in bone marrow failure syndromes.

Disclosures
Niemeyer: Celgene: Consultancy.