bluuebunny/crossref_metadata_2025_split huggingface dataset:
Children with Inflammatory Bowel Disease Exhibit Insensitivity to Tissue Factor Pathway Inhibitor
Authors: Schlagenhauf Axel , Haidl Harald , Sina Pohl , Jörg Jahnel , Muntean Wolfgang , Siegfried Gallistl 
Published: October 2019

Abstract
Background: Patients with inflammatory bowel diseases (IBD) exhibit chronic inflammation of the digestive tract associated with a prothrombotic shift in the plasmatic coagulation system as well as heightened platelet reactivity and preactivation. Recently, microbial and platelet-derived inorganic polyphosphate has been shown to inhibit tissue factor pathway inhibitor (TFPI), thus, influencing the hemostatic balance during inflammation. We hypothesized that polyphosphate plays a role in the pathophysiology of inflammatory bowel disease resulting in refractoriness to TFPI activity.
Aims: We aimed to determine the hemostatic sensitivity to exogenous TFPI in IBD patients and healthy controls.
Methods: Plasma from pediatric patients with active Crohn's disease (CD) (N=10) or ulcerative colitis (UC) (N=10) and age-matched healthy controls (N=20) was spiked with recombinant TFPI (150 ng/ml). Thrombin generation with/without exogenous TFPI was performed using Calibrated Automated Thrombography. Differences in lag time of thrombin generation with/without TFPI were calculated (∆lag time).
Results: Without addition of exogenous TFPI, IBD patients exhibited a shorter lag time than controls (IBD: 2.65±0.41 min; Controls: 3.72±0.48 min, P<0.001) Addition of exogenous TFPI prolonged the lag time significantly in healthy controls (∆lag time= 4.69±0.61 min; P<0.001), while the lag time was just slightly prolonged in IBD patients (CD: ∆lag time=0.92±0.37 min; P<0.001; UC: ∆lag time=0.34±27 min; P<0.05).
Conclusion: Plasma samples from pediatric IBD patients exhibit refractoriness towards the anticoagulant activity of exogenous TFPI. This hyposensitivity potentially extends to the action of endogenous TFPI which adds to the prothrombotic phenotype associated with IBD. Further studies are needed to determine potential associations with disease activity and the susceptibility to develop thrombosis.

Disclosures
No relevant conflicts of interest to declare.
Changing the Paradigm in Hemophilia Care: Extended Half-Life Products (rFVIIIFc and rFIXFc)
Authors: Aledort Louis M. , Jain Nisha , Li Nanxin , Krishnan Sangeeta , Hagberg Benjamin , Su Jun 
Published: October 2019

Abstract
Background:
Prophylactic treatment with factor products has demonstrated superior outcomes and care compared to on-demand treatment in hemophilia patients, including patients with pre-existing joint diseases. However, 40% to 50% of severe or moderate hemophilia A and B patients are still treated on-demand in the US. Recombinant factor VIII and IX Fc fusion proteins (rFVIIIFc and rFIXFc) were the first extended half-life (EHL) products approved in the US (in 2014) with the promise of optimizing prophylactic regimens by achieving low bleeding rates and improving joint health with fewer injections overall.
Objective:
This is the first real-world study to assess the impact of EHL rFVIIIFc and rFIXFc on overall prophylaxis rates, particularly to understand how patients have transitioned from on-demand treatment.
Methods:
A retrospective analysis was conducted on aggregate, de-identified United States Specialty Pharmacy Provider (SPP) records from Jan 2014 to June 2018 (data pull on June 14th, 2018). Adult hemophilia patients (age 18 and above) were included for an analysis of treatment regimens prior to and after switching to EHL rFVIIIFc and rFIXFc (on-demand or prophylaxis). The results were descriptively compared to those who switch to the other EHL PEGylated rFVIII and rFIX albumin Fusion Protein (rFIX-FP) and conventional factors.
Results:
A total of 304 hemophilia A patients who switched to rFVIIIFc and 133 hemophilia B patients who switched to rFIXFc were included in this study. For the hemophilia A and B patients, the median age was 31 and 32 years and median weight was 82 and 79 kg. The proportion of patients who switched from on-demand treatment with conventional factors to prophylaxis treatment with EHL rFVIIIFc and rFIXFc among all patients who switched to these products increased over time since the FDA approval (3 time points: 2014-2015, 2016-2017, and 2018: rFVIIIFc: 13%, 20%, and 21%; rFIXFc: 25%, 29%, and 57%). Such proportions were higher compared to those who switched from on-demand with conventional factors to prophylaxis with the other EHLs (2016-2017, 2018: PEGylated rFVIII: 16%, 18%; rFIX-FP: 18%, 13%, see comparison in the Figure) and with conventional factors (2014-2015, 2016-2017, and 2018: conventional rFVIII: 10%, 7%, and 9%; conventional rFIX: sample size was too small and not suitable for analysis).
Discussion:
rFVIIIFc and rFIXFc have transformed the paradigm of hemophilia care by shifting on demand patients to prophylaxis significantly faster than conventional factors and any other EHLs to date. This shift is significant as it is helping more hemophilia patients achieve optimal protection through low bleeding rates and improved joint health over the long-term.
Figure. Figure.

Disclosures
Jain: Bioverativ: Employment. Li:Bioverativ: Employment. Krishnan:Bioverativ: Employment. Hagberg:Bioverativ: Employment. Su:Bioverativ: Employment.
Impact of Non-CMV Specific Intravenous Immunoglobulin on Intravenous Ganciclovir Effectiveness
Authors: Drokov Mikhail Yu. , Tikhomirov Dmitry S. , Kuzmina Larisa , Tupoleva Tatiana A. , Vasilyeva Vera , Popova Natalia , Mikhaltsova Ekaterina , Koroleva Olga , Dubnyak Daria , Sidorova Anna , Usikova Ekterina , Konova Zoya , Dmitrova Anna , Gaponova Tatiana V. , Parovichnikova Elena , Savchenko Valery G. 
Published: October 2019

Abstract
Introduction Cytomegalovirus (CMV) reactivation is one of the main problems in allogenic hematopoietic stem cell transplantation (allo-HSCT). Standard of care for CMV-disease is still ganciclovir at 10 mg/kg b.i.d. Use of non-CMV specific intravenous immunoglobulin (IVIG) is still controversial and mostly determined by internal care protocols in every transplant center. Here we report data about impact of non-CMV specific IVIG during ganciclovir treatment in allo-HSCT CMV-patients.
Patients and methods Thirty two patients with hematological malignancies transplanted in NRCH were included in this study. Detailed patients characteristics are listed in Table 1. All patients were CMV-positive by PCR (Amplisens EBV/ CMV /HHV6-screen-FL", InterLabService, Russia) and required to start therapy with ganciclovir 10 mg/kg when was included to this study. Patients in IVIG group also received non-CMV specific IVIG during ganciclovir treatment with a median total dose 20 g (7.5-90 g.). Kaplan-Meier estimator was used to determine probability of achievement of CMV-negativity from time of ganciclovir was started to CMV-negativity (by PCR) or last contact. The logrank test is used to compare differences between two groups. Fisher's exact test was used for 2×2 tables. P-value less than 0.05 was considered statistically significant
Results Influence of IVIG in allo-HSCT patients on probability of achievement of CMV-negativity (by PCR) are shown in the Figure 1. As we can see there is no significant differences on the achievement of CMV-negativity (by PCR) in patients with/without IVIG during ganciclovir therapy.
Conclusion The use of non-CMV specific immunoglobulins after allo-HSCT still lies in the plane of faith and the possibilities of the transplantation center than any evidence base. Using IVIG in patients after allo-HSCT is still not a routine in terms of CMV-reactivation/disease therapy and mostly based on an "experience" of every transplant center. According to our data there is no difference between IVIG/no-IVIG groups during ganciclovir treatment so use of IVIG, for our opinion, does not really make sense for CMV-disease treatment. Anyway a large randomized trial is required for determine indications for therapy with IVIG.

Disclosures
No relevant conflicts of interest to declare.
Exome Sequencing of Extreme Phenotypes Suggests Novel Candidate Genes As Modifiers of Leg Ulcer in Sickle Cell Anemia
Authors: de Carvalho Siqueira Gabriela Queila , Ananina Galina , Borges Murilo Guimarães , de Souza Bruno Batista , Cendes Iscia Teresinha Lopes , Costa Fernando F. , de Melo Mônica Barbosa 
Published: October 2019

Abstract
Background: Despite resulting from homozygosity of a single mutation at position 6 of the beta-globin locus, the clinical aspects of sickle cell anemia (SCA) are very heterogeneous. Leg ulcer is one of the many resulting complications, exerting a quite negative impact in the quality of life of patients and is related to the severity of the disease. The pathogenesis of such complication is complex and still not well explained. Objective: Seeking for novel genes associated with leg ulcer in SCA, we proposed exome sequencing of extreme phenotypes in a sample of the Brazilian population. Methods: Our sample consisted of 40 unrelated patients with SCA from the Hematology and Hemotherapy Center of the University of Campinas (Unicamp), Campinas, SP, Brazil. The cohort was composed by 20 patients who didn't have leg ulcer, with ages varying from 40 to 61 years (it is unusual the appearance of this disorder at such age) and 20 patients with chronic leg ulcer. DNA was collected from peripheral blood leukocytes and submitted to exome sequencing. Capture and enrichment were performed with the Nextera Rapid Capture Exome kit (Illumina) and samples were loaded in the Illumina HiSeq2500. The bioinformatics process was based on the GATK Pipeline and manual filters in vcftools and excel. Briefly, we prioritized the variants according to some conditions, like score quality, depth, Hardy-Weinberg equilibrium and minor allele frequency greater than 0.1. Subsequently, through PLINK, we selected the variants with p value less than 0.05 from Fisher association test. The annotation was made through wAnnovar. We excluded the synonymous variants, remaining with 244 variants at this point. Finally, the deleterious variants were selected, in addition to those not classified by SIFT, PolyPhen2 and FATHMM. Twenty-one variants remained, all of them in genes beyond the usual set of a priori biological candidate genes for this phenotype. We picked up 6 variants with the lowest p values: rs4857302 (CRYBG3), rs3782489 (KRT77), rs11800462 (TNFRSF25), rs13428956 (FOXD4L1), rs201853154 (UBTFL1) and rs11454536 (VWDE). We suggest that these variants and their genes could potentially be related to the development of leg ulcer in SCA. However, they should all be validated in other populations for confirmation.

Disclosures
No relevant conflicts of interest to declare.
Identification of Syndecan-1 As a Key Dependency of Myeloid Leukemia Growth and Dissemination
Authors: Spinler Kyle , Reya Tannishtha , Bajaj Jeevisha , Ito Takahiro , Zimdahl Bryan , Ahmadi Armin , Koechlein Claire , Lytle Nikki , Kwon Hyog Young , Anower-E-Khuda Ferdous , Kritzik Marcie , Karlseder Jan , Park Pyong Woo , Esko Jeffrey 
Published: October 2019

Abstract
Intratumoral heterogeneity is a common feature of many myeloid leukemias and a significant reason for treatment failure and relapse. Thus identifying the cells responsible for residual disease and leukemia re-growth is critical to better understand how they are regulated. Here we show that a knock-in reporter mouse for the stem cell gene Musashi 2 (Msi2) allows identification of therapy resistant leukemia propagating cells in aggressive myeloid malignancies, and provides a new strategy for defining their core dependencies. Specifically, we carried out a high throughput screen using Msi2 reporter blast crisis chronic myeloid leukemia (bcCML) and identified syndecan-1 (Sdc1), a cell surface proteoglycan, as preferentially expressed in therapy resistant bcCML cells, and critical for bcCML function. Specifically, in Sdc1-/- mice, Sdc1 loss led to a defect in bcCML growth and propagation in vitro and in vivo, and markedly improved survival. Further, live imaging revealed that Sdc1 loss had a striking impact on the spatiotemporal dynamics of leukemia cells, impairing their localization, migration and systemic dissemination. Mechanistically, distinct elements of Sdc1 contributed to leukemia growth and dissemination, with the core protein alone being able to rescue the growth defect, but the heparin sulfate chains that mediate matrix attachment being needed for migration. These data present a new platform for delineating the biological underpinnings of leukemia stem cell function, and identify Sdc1 as a central regulator of leukemia stem cell growth and dissemination.

Disclosures
No relevant conflicts of interest to declare.
Relationship of Markers of Inflammation, Infection and Endothelial Function to Mortality and Severity of Coagulopathy in Patients with Sepsis-Associated DIC
Authors: Hoppensteadt Debra , Walborn Amanda , Rondina Matthew T. , Fareed Jawed 
Published: October 2019

Abstract
Background: Sepsis-associated disseminated intravascular coagulation (DIC) is a complex clinical scenario involving derangement of many processes, including hemostasis. Assessment of markers of these other aspects of disease, including inflammation, immunity, endothelial function, and endogenous anticoagulants may provide insight into DIC pathophysiology and lead to improved methods for assessment of patient condition and response to treatment. This study was designed to measure biomarkers representative of multiple systems involved in DIC development in a cohort of patients with sepsis and DIC and determine the association of these biomarkers with disease severity and mortality.
Materials and Methods: Citrated plasma samples were prospectively collected from 102 patients with sepsis and suspected DIC at ICU admission (day 0) and again serially on ICU days 4 and 8 under an IRB approved protocol. DIC score was determined using the ISTH scoring algorithm (e.g. platelet count, PT/INR, fibrinogen, and D-Dimer). Plasma from healthy individuals was purchased from a commercial laboratory (George King Biomedical, Overland, KS). . CD 40 Ligand (CD40L), Plasminogen inhibitor 1 ( PAI-1), nucleosomes, Procalcitonun (PCT), Microparicle Tissue Factor (MP-TF), and Prothrombin Fragment 1.2 (F1.2) were measured using commercially available ELISA kits. Protein C activity was measured using a clot-based assay. Interleukin 6 (IL-6), Interleukin 8 (IL-8), Interleukin 10 (IL-10) , Tumor Necrosis Factor Alpha (TNFα) , Monocyte Chemoattractant protein 1 (MCP-1) were measured using a biochip technology.
Results: Significant differences in levels of Protein C (p=0.009), PCT (p=0.0005), IL-6 (p=0.019), IL-8 (p= 0.015), and PAI-1 (p=0.015) were observed between survivors and non-survivors (Table 1). Significant variation of Protein C (p=0.002), nucleosomes (p=0.05), PCT (P<0.001), IL-6 (p=0.001), IL-8 (p=0.003), IL-10 (p=0.011), TNFα (p=0.021), and MCP-1 (p=0.21), were observed based on severity of DIC score.
Conclusion: Markers from multiple systems perturbed in DIC were associated with mortality suggesting that while these systems may not be routinely evaluated in the course of patient care, dysfunction of these system contribute significantly to mortality. In addition, numerous inflammatory cytokines showed an association with DIC score. This study suggests that the measurement of additional markers in sepsis-associated DIC may be of value in the prediction of mortality and may be helpful in guiding treatment for these patients.

Disclosures
No relevant conflicts of interest to declare.
Role of Point-of-Care Device in Improving Sickle Cell Newborn Screening Feasibility in Haiti
Authors: Alvarez Ofelia A. , Hustace Tally , Voltaire Mimose , Rizil Rodrigueson , Liberus Ulrick , Saint Fleur Rony 
Published: October 2019

Abstract
Introduction: Until 2017, hemoglobinopathy newborn screening (NBS) was not offered in Haiti, a country with about 243,000 births per year. Sickle SCAN is a rapid point-of-care (POC) with lateral flow immunoassay technology, but its role in NBS is unknown. Recently, over 100 dried blood samples from newborns were tested [Nguyen-Khoa T, Ann Biol Clin (Paris), 2018] in France with accurate results. Objective: This work had the objective to be proof-of-concept that a hospital-based NBS program is feasible in Haiti and to examine the role of Sickle SCAN in NBS in a larger sample size. Methods: The Ethics Committee at the Universitaire Justinien Hospital (HUJ) and the Institutional Review Board at the University of Miami approved this study. We formed a Haitian team composed of a pediatrician on staff (RSF), two dedicated NBS nurses, a program coordinator (MV), two community health workers, a laboratory technician (RR), and a data manager (UL). We acquired isoelectric focusing equipment (IEF) and performed NBS from dried blood samples only for the first three months while the technician gained proficiency. Thereafter, we have performed dual screening method with the POC device Sickle SCAN and IEF. Confirmatory testing was obtained with both methods. Data were entered into RED Cap. Results: Beginning in August 2017 until present (June 2018), we have screened 1,800 newborns, of which 10.5% have sickle cell trait, 3.3% have hemoglobin C trait, 0.78% have hemoglobin SS, 0.27% have hemoglobin SC and one child has been confirmed to have sickle beta thalassemia plus. Currently, there are 15 children followed at HUJ, for a SCD incidence of 0.83% (8 SS, 6 SC and 1 S-beta thalassemia+). Before the POC screening was implemented two infants have already died when the mothers were contacted (one who screened positive for FS and another for FC). The POC allows for immediate referral and penicillin prophylaxis for at-risk children until the cases are confirmed. Systemic barriers encountered are the electrical outages and intense heat which impacted IEF performance, material procurement (laboratory materials, NBS materials, oral penicillin) which are dependent of shipments from outside of Haiti.
Conclusions: A hospital-based NBS Program is feasible. SCD is highly prevalent with an incidence of 0.83% among newborns in the population studied. Based on this incidence, we estimate that every year around 2,000 children will be born with sickle cell disease in Haiti. The point-of-care device enhanced the screening program by obtaining immediate screening results and maximizing family notification for confirmatory testing.

Disclosures
No relevant conflicts of interest to declare.
Expression Status of Prohibitin Gene Predicts the Prognosis of Acute Myeloid Leukemia with Normal Cytogenetics
Authors: Shin Myung Geun , Lee Jun Hyung , Kim Hye Ran , Na Bo Ram , Lee Young Eun , Choi Hyun Jung , Kim Soo Hyun , Shin Jong Hee , Suh Soon Pal 
Published: October 2019

Abstract
Introduction: Cytogenetically normal acute myeloid leukemia (CN-AML) accounts for 40 to 50 percent of all AML and shows significant heterogeneity in clinical outcomes. Despite the advancements in the molecular pathophysiology of CN-AML, clinical outcome of this leukemia remains to be elucidated. Prohibitin (PHB) were typically known to have the potential act as a tumor suppressor, anti-proliferative regulator of cell-cycle in normal cells. However, our previous study discovered that high expression of PHB in AML cells was involved in escaping apoptosis process. In this study, we investigated the clinical significance of expression levels of PHB1 and PHB2 genes in patients with CN-AML.
Methods: We developed quantitative reverse transcription PCR (qRT-PCR) detection system for mRNA expression status for PHB1 and PHB2.The expression levels was analyzed in 69 CN-AML patients. Total RNA was extracted from bone marrow samples at the diagnosis using InvitrogenRNAqueous Total RNA Isolation Kit (Thermo Fisher Scientific, USA). The concentration of isolated total RNA was measured by Micro UV-VIS spectrophotometer (Malcom, Japan). Quantification of mRNA of PHBs were performed in CFX96 Touch Real-Time PCR system (BIO-RAD, USA) using our laboratory-developed, customized primers and probes.The gene expression levels werenormalized compared to those of ABL1 gene.Survival analysis was performed compared with other prognostic factors: age, NPM1 mutation, and FLT3-ITD. FLT3-ITD was tested by PCR & fragment analysis (LeukoStrat FLT3 Mutation kit; Invivoscribe, USA) and NPM1 mutation was tested using qRT-PCR kit (ipsogen NPM1 MutaQuant; Qiagen, Germany). All statistical computations and graphical process were performed using R version 3.5.1. A P value ≤ 0.05 (for 95% confidence interval) was considered statistically significant.
Results: The median age of enrolled patients was 68 (ranged 23-85) at the diagnosis. The ratio of male and female was 1:0.86 (37:32). There was no significant difference in age, sex, WBC count in peripheral blood, blast percentage of peripheral blood and bone marrow, the achievement rate of complete remission, and the enforcement of stem cell transplant according to expression status of PHB1 and PHB2. Using 0.61 as cutoff, PHB1 high-expressed group showed superior overall survival (OS) (47.7% vs 15.1%; P = 0.014) and low hazard ratio (0.43 [95% CI: 0.215-0.861]; P = 0.017) compared to PHB1 low-expressed group. In PHB2, using a cutoff 2.39, high-expressed group showed as inferior OS (17.5% vs 40.0%; P = 0.16) and high hazard ratio (1.93 [95% CI:0.739-5.042]; P = 0.18) compared to low expressed group.
Conclusions: PHB1 expression status was associated with favorable clinical outcome. In contrast, those of PHB2 was related with poor prognosis in CN-AML.

Disclosures
No relevant conflicts of interest to declare.
Treatment of Epstein-Barr Virus Associated Central Nervous System Diseases after Allogeneic Hematopoietic Stem Cell Transplantation with Intrathecal Donor Lymphocyte Infusion
Authors: Zhao Juanjuan , Zu Yingling , Han Lu , Zhang Yanli , Gui Ruirui , Yu Fengkuan , Li Zhen , Zhao Huifang , Fang Baijun , Lin Quande , Song Yongping , Liu Delong , Zhou Jian 
Published: October 2019

Abstract

Allogeneic hematopoietic stem cell transplantation(allo-HSCT) is an effective measure for the treatment of hematological disease. With the progress and widespread use of allo-HSCT, Epstein-Barr virus(EBV) related central nervous system(CNS) diseases have gotten more and more attention because of its poor prognosis and overall survival. Since currently there is no standard treatment for patients with EBV-associated CNS diseases and reported therapies are heterogeneous with mixed results, we attempted to develop a novel therapeutic method. We have applied a regimen of intrathecal donor lymphocyte infusion(IDLI) in 3 patients with EBV-associated CNS diseases after allo-HSCT in addition to immunosuppressants reduction and combined antiviral therapy. All of 3 patients were responsive to this therapy: all clinical symptoms and EBV load in CSF resolved 10, 17 and 12 days after initial IDLI respectively, and magnetic resonance imaging (MRI) showed lesions of case 1 and 2 disappeared 15 and 19 days after initial IDLI respectively. Even more appealing, there were no acute or chronic adverse reactions during the infusion and up to 23 months of follow-up. In conclusion, IDLI seems to be an effective and safe method for EBV-associated CNS diseases in allo-HSCT recipients. We recommend this treatment modality for further investigation.

Disclosures
No relevant conflicts of interest to declare.