bluuebunny/crossref_metadata_2025_split huggingface dataset:
Age-Related Changes in Methylome and Transcriptome of Hematopotietic Stem Cells Underlie Natural Genetic Variations
Authors: Liang Ying 
Published: October 2019

The aging of hematopoietic stem cells (HSCs) contributes to the aging of blood system and perhaps the whole organism. The aging process is coordinately determined by both genetic and epigenetic factors, and demonstrates inter-individual variations. We used high-throughput sequencing methods to study the age-dependent changes of genome-wide DNA methylation and gene expression patterns in HSCs of C57BL/6 (B6) and DBA/2 mouse strains, which have shown natural variations in HSC aging process. We observed global age-associated decrease of DNA methylation in both strains, but D2 HSCs have a stronger loss of epigenetic control than B6 stem cells during aging. Majority age-related changes of DNA methylation occur from young to mid-aged stages. We identified stable strain-specific differentially methylated regions (DMRs) that overlap with cis-eQTLs. Moreover, transcription factor binding site motifs are more likely to be disrupted in the DMRs, suggesting the potential impact of genetic variations on epigenetic regulation of HSC aging. We further demonstrated that strain-specific DMRs have more profound effects on the aging of B6 HSCs than D2 stem cells. Transposons are differentially regulated by the DMRs in the two strains, in which D2 HSCs are prone to transposon insertion. This study comprehensively investigated the effects of natural genetic and epigenetic variations on HSC aging. Loss of DNA methylation is an epigenetic signature of stem cell aging, and DNA methylation variations correlates with genetic variations, both contributing to inter-individual differences in stem cell and perhaps organismal aging.

Disclosures
No relevant conflicts of interest to declare.
Autologous Hematopoietic Cell Transplantation in Patients with Multiple Myeloma: IMPACT of Age
Authors: Dhakal Binod , Nelson Ariel , Guru Murthy Guru Subramanian , Fraser Raphael , Eastwood Daniel , Hamadani Mehdi , D'Souza Anita , Hari Parameswaran 
Published: October 2019

Abstract
Background: In both the novel and pre-novel agents era, high dose therapy followed by autologous hematopoietic cell transplant (AHCT) has been shown to prolong survival in multiple myeloma (MM) in randomized trials, but prospective have only included younger patients (65 -70 years and younger). Given that the median age at diagnosis of MM is 66 years, it is important to know the outcomes of AHCT in older patients. Similarly, the definite outcomes of AHCT in very young patients (<50 years) is also lacking as they represent a very small proportion in clinical trials.
Methods: We analyzed a consecutive cohort of patients with MM receiving ASCT from 2000-2015 in two different age groups, old (>70 years; N=105) and young (≤50 years, N=86) and compared the outcomes. The primary objectives were to assess overall survival (OS), progression free survival, (PFS), and non-relapse mortality (NRM) in these two groups.
Results: Eighty-sex patients were young (≤50 years), while 105 patients were old (>70 years). Young patients had better performance status and lower co-morbidity index, while majority of older cohort received dose reduced high dose melphalan between 140-180 mg/m2. Median follow up of survivors in the younger cohort was 33 months (range, 2-164) compared to the 22.5 months (3-133) in the old group (p=0.02). The PFS at 1 year was 60% (95%CI 46-72) for young patients and 58 (95%CI, 45-69) for the old ones. The OS at 1 year was 92% (95% CI 84-96) for younger and 85% (95% CI 76-91) for the older cohorts. On multivariate analysis, age did not have any effect on survival (p =0.82), high-risk cytogenetics (HR 2.2, 95% CI 1.06-4.6, p=0.04) was associated with higher mortality. High-risk cytogenetics (HR 1.2, 95% CI 1.1-3.5, p=0.004) and non-responding or progressive disease at transplantation (HR, 5.0 95% CI 1.8-13.5, p=0.02) were significantly associated with inferior PFS.
Conclusion: Age by itself should not be a limiting factor in considering the modality of AHCT. However consideration could be given to augmenting therapy for young patients with additional novel treatments post transplant given their similar outcomes with the old patients based on our study.
Overall survival between the two age groups, 1: >70 years old and 2: ≤50 years old. Overall survival between the two age groups, 1: >70 years old and 2: ≤50 years old. Figure Figure.

Disclosures
Hamadani: Takeda: Research Funding. Hari:Merck: Research Funding; BMS: Honoraria.
Safety and Efficacy of Direct-Acting Antiviral Drugs in Patients with Haemoglobinophaties and Chronic Hepatitis C Infection
Authors: Piga Antonio , Rigano Paolo , Origa Raffaella , Cappellini M. Domenica , Pinto Valeria , Tartaglione Immacolata , Filosa Aldo , D'Ascola Domenico , Di Marco Vito , Forni Gian Luca 
Published: October 2019

Abstract
Background and Aim: Direct-acting antiviral drugs (DAAs) have a very high efficacy in patients with hepatitis C virus (HCV) infection, but they have not been extensively used in patients with haemoglobinophaties. To evaluate the safety and efficacy of DAA regimens in this subset we used the ITHACA-SITE dataset, which includes patients with haemoglobinophaties and chronic HCV liver disease treated in Italy.
Patients and methods: Between March 2015 and June 2016, 121 patients included in the ITHACA-SITE dataset started DAA regimens. Cirrhosis was defined by FibroScan®showing≥12 kPa performed within 6 months before the treatment. Regimen choice and use of ribavirin were based on viral genotype and stage of disease, according to guidelines. Negative HCV RNA at week 12 of post-treatment follow up was considered as Sustained Virological Response (SVR).
Results: The mean age of 121 patients was 42 years, 75 (62%) were males, 101 (83.5%) had β-thalassemia major, 9(7.5%) had β-thalassemia intermedia and 10 (8.3%) had sickle cell disease. Sixty-six patients (54%) had the diagnosis of chronic hepatitis and 55 (46%) hadthe diagnosis of cirrhosis. The prevalence of HCV genotypes (G) was: G1a 6 (5%), G1b 70 (57.8%), G2 31 (25.6%), G3 6 (5%), G4 8 (6.6%). Fifty-six patients (46.3%) were Peg-interferon (P) and Ribavirin (R) naïve and 65 (53.7%) were P/R experienced, 20 patients (16.5%) had diabetes and 29 (24%) had heart disease. By June 2016 63 patients (52%) had concluded the treatment and 33 (27.3%) had concluded the post-treatment follow-up. By ITT, the rate of response at the end of treatment (ETR) was 100% (63/63) and 94% (31/33) of patients achieved a SVR. No patients stopped treatment because of adverse events. No interference with chelation therapy was observed.
Conclusions: Use of DAAs regimens in practice is safe and effective in patients with haemoglobinophaties and cirrhosis or chronic hepatitis due to HCV. The therapy is indicated also in patients with co-morbidity. The lifetime utility of HCV eradication in terms of reduction of events and overall survival needs evaluation in long-term observational cohorts.

Disclosures
Piga: Novartis: Research Funding; Apopharma: Honoraria. Di Marco:Gilead: Research Funding. Forni:Novartis, Celgene: Research Funding.
Endothelin-1 Regulates Molecules of the Major Histocompatibility Complex: Role in Sickle Cell Disease
Authors: Rivera González Lorena , Inostroza-Nieves Yaritza , Lozano Alexandra , López Pablo J. , Rosado Alicea Jamie , Prado Gregory N. , Romero Jose R. , Rivera Alicia 
Published: October 2019

Abstract
Molecules of the Major Histocompatibility Complex (MHC), and in particular specific human leukocyte antigen (HLA) alleles, have been proposed in the pathophysiology of immune and vascular alterations leading to vasoocclusive crises (VOC) and stroke in Sickle Cell Disease (SCD). Endothelial cells express MHC molecules following exposure to cytokines. SCD is characterized, in part, by vascular endothelial cell activation, increased oxidative stress, sickle cell adhesion and excess levels of the potent mitogen, endothelin-1 (ET-1). ET-1 activates endothelial cells, induces oxidative stress and inflammation in the vascular wall and regulates erythrocyte homeostasis. However, the role of ET-1 on MHC regulation in SCD is not clear. We first characterized the effect of ET-1 on MHC expression in the human endothelial cell line, EA.hy926. We observed dose-dependent increases in the expression of MHC class I (HLA-A2 4.8 ± 2.1 folds p<0.01 n=4), MHC class II (HLA-DR 4.4 ± 1.7 folds p<0.01 n=4) and MHC transcription factor (CIITA 3.5 ± 1.8 folds p<0.05 n=4) in EA.hy926 cells. ET-1-stimulated expression of HLA-A2, HLA-DR and CIITA were significantly blocked by pre-incubation of cells with 10 µM BQ788, a selective blocker of ET-1 type B receptors (p<0.01, n=4). Chromatin immunoprecipitation (ChIP) studies in EA.hy926 cells showed that ET-1 increased Histone H3 acetylation of the promoter region within MHC molecules (HLA-A2 62% ± 5%, HLA-DRB 33% ± 18%, p<0.01, n=4); an event that was likewise blocked by BQ788. We then studied two sickle transgenic knockout mouse models of moderate to severe disease phenotype, βSAntilles and Berkeley (BERK) mice, respectively. We observed significant increases in MHC molecule, H2-Aa mRNA levels (n=7; p<0.01) in spleens from sickle transgenic mice when compared to transgenic knockout mice expressing human hemoglobin A (HbA). We then treated BERK, βSAntilles and HbA mice for 14 days with ET-1 receptor antagonists and observed significant reductions in H2-Aa mRNA levels in spleen tissue from sickle transgenic mice but not in HbA mice (n=7; p<0.05). These results implicate ET-1 as a novel regulator of MHC molecules and suggest that ET-1 receptor blockade represents a promising therapeutic approach to regulate both immune and vascular responses in SCD.

Disclosures
No relevant conflicts of interest to declare.
Increased T Follicular Helper Cells in Patients with Aplastic Anemia
Authors: Verigou Evgenia , Galanopoulos Athanasios , Kattamis Antonios , Symeonidis Argiris , Solomou Elena E. 
Published: October 2019

Abstract
Introduction:T-follicular helper cells represent a sub-population of CD4+helper T cells (TFH), and they are characterized by the surface expression of CXCR5, ICOS, and PD1, the transcription factor Bcl-6, and produce mainly IL-21, but also IL-17, IL-4, and IFN-γ. They represent the major population that helps B cells to turn into plasma cells and they are implicated in the pathogenesis of different autoimmune diseases. Peripheral CXCR5+CD4+ T helper cells (p-TFH) are the circulating component of TFH. p-TFH cells have been extensively studied in the context of inflammation and autoimmunity. Patients with systemic lupus erythematosus and rheumatoid arthritis have increased p-TFH. T follicular regulatory T cells (Tfr) express CXCR5, ICOS and PD1, CD25 but also FOXP3, and they can inhibit B cell responses.
Immune dysregulation characterizes patients with aplastic anemia. We and others have previously shown that aplastic anemia patients have decreased levels of regulatory T cells (Tregs) at presentation and increase if they respond to treatment. Additionally, these patients display increased Th17 cells at presentation.
Specific aim: In this study we wanted to examine the expression of p-TFH and Tfr cells in patients with aplastic anemia. We isolated peripheral blood mononuclear cells (PBMCs) from patients with aplastic anemia (n=8) and ten healthy, age- matched controls. Written informed consent was obtained from all study subjects. PBMCs were left untreated or activated with PMA and Ionomycin, in the presence of Brefeldin A, for 5 hours. Subsequently, cells were stained with the surface markers CXCR5, CD4, CD57, CD25, CD127, CD278, CD279, IFN-g,IL-21, and IL-17, and subsequently analyzed by flow cytometry.
Results: Patients with aplastic anemia showed increased expression of CD4+CXCR5+ cells (Tfh) compared to healthy controls (3,03±0,55% vs 1,3±0,52% respectively, p=0.08). Additionally, these patients also showed decreased expression of Treg cells compared to healthy controls (0,74±0,27% vs 2,25±0,70% respectively, p=0.03). When analyzed for Tfr cells, patients with aplastic anemia showed minimal levels of these cells compared to helathy controls (0,05±0,02% vs 1,14±0,16% respectively, p<0.001) (Fig. 1).
Conclusion: Although the number of patients analyzed is limited, our preliminary results suggest that aplastic anemia patients show increased CXCR5+CD4+ cells compared to age-matched control subjects, and decreased Tfr cells. Further analysis of a larger pool of subjects is underway along with the examination of the specific transcription factor Bcl-6, to reveal the role of p-TFH and Tfr cells in the immune pathogenesis of aplastic anemia.
Figure Figure.

Disclosures
No relevant conflicts of interest to declare.
Assessing Patient Preferences for the Benefits and Risks of Treating for Acute Myeloid Leukemia (AML): A Pilot Study
Authors: Seo Jaein , Smith B. Douglas , Estey Elihu H. , Voyard Ernest S. , O'Donoghue Bernadette , Bridges John F. P. 
Published: October 2019

Abstract
Introduction:Acute Myeloid Leukemia (AML) is a blood cancer which progresses rapidly in the absence of treatment. There have been few advances in the treatment of AML over the last three decades. In 2015 the Leukemia and Lymphoma Society (LLS) initiated a research program to assess patient preferences for AML treatments. The aim of this research was to promote patient-focused drug development and inform future regulatory decisions. We sought to develop and pilot a patient-centered survey instrument to assess patient preferences for the benefits and risks of AML treatments.
Methods: Development was informed by a targeted literature review and engagement with an expert stakeholder committee (n=12) to guide the clinical accuracy and relevance of the survey instrument. A community stakeholder committee, consisting of patients with AML and caregivers (n=15), provided information about their experiences with AML and various treatments. They also engaged in pretest interviews to test comprehension and ensure it captured the patient experience. A discrete-choice experiment (DCE) was developed spanning 5 benefits and risks, including event-free survival (EFS), complete remission (CR), time in hospital, short-term side effects, and long-term side effects. This DCE consisted of 16 pairs of hypothetical treatments, with participants being asked to identify which treatment they would prefer in each pair. Results of a pilot study with AML patients and caregivers were assessed by Z-score that were derived from a conditional logistic model regressing each attribute upon their choices.
Results: The pilot included 18 patients and 8 caregivers with a mean age of 50 years (range=24-81). Most participants were college educated (n=22), Caucasian (n=19), privately insured (n=21), and employed (n=13). Participants valued CR the most (Z-score=7.95, p<0.001), followed by EFS (5.32, p<0.001). They were most averse to time in hospital (-3.41, p=0.001), followed by long-term side effects (-3.03, p=0.002) and short-term side effects (-1.99, p=0.047), which was marginally significant.
Conclusions: This study demonstrates the value of rigorous community engagement in developing survey instruments to measure patient preferences. The results of this pilot study demonstrate the ability of our DCE to measure treatment preferences of AML patients and caregivers. Given this success, we are currently engaged in a nationally study where we will recruit a larger and more diverse sample. The results of this national study will inform drug developers and regulatory decision makers.

Disclosures
Seo: The Leukemia & Lymphoma Society: Research Funding. Voyard:The Leukemia & Lymphoma Society: Employment. O'Donoghue:The Leukemia & Lymphoma Society: Employment. Bridges:The Leukemia & Lymphoma Society: Research Funding.
Does Normal Reticulocyte Hemoglobin Rule out Iron Deficiency As a Cause of Normocytic Normochromic Anemia?
Authors: Yu Zhou , Blankenship Leann Michelle , Ibrahim Mohammed , Gbadamosi Bolanle , Stender Michael , Anderson Joseph , Jaiyesimi Ishmael , Huang James 
Published: October 2019

Abstract

Background: Normochromic, normocytic anemia is the most frequently encountered type of anemia with a wide differential diagnosis. Iron studies are frequently ordered as part of the anemia work up as iron deficiency anemia could be potentially normocytic and normochromic. Recently reticulocyte hemoglobin has emerged as a marker to help identify early stages of iron deficiency at a time that other traditional biochemical studies for iron deficiency are non-informative. We hypothesize that normal reticulocyte hemoglobin level is a sensitive negative predictor of iron deficiency.
Aim:To evaluate whether a normal reticulocyte hemoglobin level can be used to exclude iron deficiency anemia in patients with normocytic normochromic anemia.
Methods:We reviewed the results of all hematological tests obtained as a part of hematopathology consults which were requested between January to April of 2016. Electronic medical records were screened for inclusion criteria. We included patient with hemoglobin less than 12g/dl, normal MCV (80-100) and normal reticulocyte hemoglobin level (>31 pg). We calculated the percentage of patients who had normal iron store suggested by ferritin, total iron binding capacity and iron saturation studies.
Results:We included 41 patients who has normocytic normochromic anemia with normal reticulocyte hemoglobin level. 39 out of 41 patients have normal iron studies indicating iron deficiency anemia was not the cause of anemia. The overall negative predictive value is 95.1%. Among the two patients whose ferritin levels are mildly decreased, suggesting possible iron deficiency, one patient had a blood transfusion prior to checking reticulocyte hemoglobin level. The other patient donates blood frequently and was started on oral iron supplementation trial without any hemoglobin improvement after two weeks. It appeared that these two patients might not have true iron deficiency.
Conclusion: Measurement reticulocyte hemoglobin level can serve as a useful marker to rule out iron deficiency as a cause of normocytic normochromic anemia. Serum Iron and ferritin studies are not indicated in patients with normocytic normochromic anemia with normal reticulocyte hemoglobin.

Disclosures
No relevant conflicts of interest to declare.
Clinical Characterization of Staphylococcus Septicemia-Associated Hemophagocytic Lymphohistiocytosis
Authors: Qiu Hongxia , Song Meng , Ni Fang , Wang Jujuan 
Published: October 2019

Abstract
Abstract Objective: To analyze the epidemic characters, pathogenesis, clinical manifestations, laboratory data, therapeutic response, and prognosis in patients with staphylococcus septicemia-associated hemophagocytic lymphohistiocytosis(HLH). Methods A r
Objective To analyze the epidemic characters, pathogenesis, clinical manifestations, laboratory data, therapeutic response, and prognosis in patients with staphylococcus septicemia-associated hemophagocytic lymphohistiocytosis(HLH).
Methods: retrospective study was performed on 7 patients with staphylococcus septicemia-associated HLH who were admitted to The First Affiliated Hospital of Nanjing Medical University from January 2010 to December 2014.
Results Seven out of 633patients(1.11%) with staphylococcus septicemia were confirmed to have HLH. The incidence of HLH in staphylococcus subspecies were:S.caprae, 12.50%;S.warneri, 12.50%;Methicillin-resistant Staphylococcusaureus(MRSA), 3.23%;S.hominis, 1.40%;S.haemolyticus, 1.25%;S.epidermidis, 0.59%, respectively. The main clinical manifestations included persistentfever(100%) and splenomegaly(85.71%). The most prominent laboratory abnormalities werehemocytopenia, abnormal hepatic function, and elevated ferritin. Bone marrow aspirations were performed on 6 patients and all of them hadhemophagocytosis shown in bone marrow smears. Among 7 patients, 4 patientssurvived(57.14%) and 3 patients died(42.86%). All the 4 cured patients were treated with antibiotic plus corticosteroid. And 2 of them used gamma globulin, while one used the COP chemotherapy regimen. Treatment with antibiotic plus corticosteroid was effective in our study.
Conclusion Treatment with antibiotic plus corticosteroid was recommended for staphylococcus septicemia-associated HLH, and gamma globulin may also be useful in reducing the risk of death.

Disclosures
No relevant conflicts of interest to declare.
Epidemiology and Management of Immune Thrombocytopenia: A Nationwide Population-Based Study in Korea
Authors: Lee Ji Yun , Lee Ju Hyun , Lee Jung-Ok , Bang Soo-Mee 
Published: October 2019

Abstract
The epidemiology of immune thrombocytopenia (ITP) is not well characterized in the general Korean population. ITP patients from 2010 to 2014 were identified using the Korean Health Insurance Review and Assessment Service (HIRA) database. Patients with diagnosis codes for secondary thrombocytopenia or secondary ITP were excluded if those codes coexisted with ITP code of D693.0 or D693.8. The prevalent cases before 1st July 2010 and incident cases from 1st July 2014 were also excluded to calculate the exact incidence of ITP. Overall age-standardized incidence rates of ITP were 5.42/100,000 person-years (95% CI: 5.33-5.51; women, 6.17 [95% CI, 6.03-6.30]; men, 4.69 [95% CI, 4.57-4.81]).The overall incidence rates of ITP appeared to have a bimodal distribution, with peaks among children (< 10 years) and in those > 70 years of age. Among adults, the age-specific incidence rates increased with age for man, but those were observed to be constant for women (figure 1). Overall, 75.2% of patients needed treatment with the median time from the diagnosis to treatment of 1.7 months (range, 0-53.3 months). First-line therapy consisted of steroids and/or immunoglobulin in 83.6%, immunoglobulin only in 14.5%, and other immunosuppressive agents such as cyclosporine A, azathioprine, mycophenolate mofetil, and danazol in 1.9%. Only 1.0% of patients underwent splenectomy and laparoscopic procedure accounted for 90.5% of these 108 patients. The median age was 46 years (range, 9-77 years) at the time of splenectomy, and 77.8% of patients were female. 58.8% of patients received treatment beyond 6months after initial therapy. Among these patients, the most frequently used therapy was steroid and/or immunoglobulin in 92.6% followed by other immunosuppressive agents in 6.9% and immunoglobulin in 0.5%. Platelet transfusions were given 13.7% of patients and the median numbers of platelets transfused were 12 units (range, 1-2,484 units) during study periods. The usage of thrombopoietin receptor agonists or rituximab couldn't be analyzed because these drugs have not been reimbursed for the treatment of ITP before 2016 in Korea. This population-based study firstly describes the incidence rates of ITP and treatment reality for patients with ITP in Korea.
Figure Age/gender-specific incidence of ITP in Korea during the period from mid-2010 to mid-2014. Figure. Age/gender-specific incidence of ITP in Korea during the period from mid-2010 to mid-2014.

Disclosures
No relevant conflicts of interest to declare.