bluuebunny/crossref_metadata_2025_split huggingface dataset:
Improving the Appropriate Use of Intravenous Immunoglobulin in the Saskatoon Health Region
Authors: Manna Mita , Karim Safiya , Dallas Karen L , Brose Kelsey 
Published: October 2019

Abstract
Intravenous Immunogloblulin (IVIG) is a plasma protein used in the treatment of various diseases. The cost of IVIG is approximately $50-$80 per gram. In 2005, Saskatchewan's usage of IVIG was above the national average. There are several published guidelines that outline recommended uses of IVIG. These are largely based on case studies and expert opinion. Despite these guidelines, IVIG is often used in multiple settings where there is no supporting evidence.
In 2013, we conducted a retrospective study on IVIG usage and found that the Saskatoon Health Region (SHR) adhered poorly to published guidelines. Following this, we aimed to determine IVIG usage in the Hematology and Neurology departments after implementing education sessions and order forms. We conducted six education sessions from December 2012 to January 2014 in the SHR. We also circulated IVIG order forms outlining appropriate indications for IVIG on the neurology and heme-oncology ward from January 1, 2014 to March 31, 2014. We then conducted a retrospective chart review of patients who received IVIG from January 1, 2014 to March 31, 2014 and compared the usage to patients who received IVIG in 2011 and 2012. Our primary endpoint was any improvement in the appropriate usage. Our secondary endpoint was adverse reactions seen in patients receiving IVIG. Statistical analysis was conducted with the use of two sample z-test for proportions.
Our results (Figure 1) showed that for hematological conditions, the proportion of patients treated appropriately increased from 0.61 in 2011 to 0.85 in the first three months of 2014 (P value = 0.05). For neurological conditions, the proportion of patients treated appropriately declined from 0.85 in the first three months of 2012 to 0.72 in the first three months of 2014 (P value = 0.52). No adverse reactions were reported.
In conclusion, our results show that after our interventions, there was significant improvement in the appropriate use of IVIG for hematologicalconditions compared to 2011. However, there was a decrease in appropriate use of IVIG for neurological conditions. Further analysis and larger sample size is required to determine the overall significance of our interventions.
Figure 1. IVIG use in Hematology and Neurology after interventions implemented Figure 1. IVIG use in Hematology and Neurology after interventions implemented

Disclosures
No relevant conflicts of interest to declare.
Age-Related Alterations in Human Hematopoietic Stem and Progenitor Cells
Authors: Rundberg Nilsson Alexandra , Bryder David , Pronk Cornelis J.H. 
Published: October 2019

Abstract
Aging of the human hematopoietic system is typically associated with various disease states and functional changes, such as decreased bone marrow (BM) cellularity, reduced adaptive immune responses and increased incidence of anemia. Similar observations are made in mice and have been shown to, at least in part, have their origin already at the hematopoietic stem cell (HSC) level. Current ideas on age-related alterations of the hematopoietic system are to a large extent based on studies of the murine system, as studies on the human system are few and to some extent contradictory. We further detailed human hematopoietic aging by comparing adult BM samples from young and old individuals, as well as samples obtained from cord blood (CB). We observed similar changes to those observed in mice, including a functional decline in the proliferative and cloning potential of HSCs and impaired lymphoid cell production with age. We revealed alterations in the frequencies of immunophenotypically defined HSCs and progenitor cells of the megakaryocytic/erythroid, granulocyte/macrophage and lymphoid lineages within the CD34+ population, consistent with the observed HSC impairment and decreased lymphopoiesis. Transcriptome profiling demonstrated a myelo-meg-erythroid bias and downregulation of genes associated with lymphoid specification in the adult aged HSC compartment compared to the young. Further, single cell gene expression analysis of 45 individual genes, including lineage affiliated genes and a panel of transcription factors, in a large number of HSCs isolated from young and old BM and CB revealed heterogeneity within the HSC compartment as well as a pattern of transcription fluctuation for a set of genes across age boundaries. Collectively, we demonstrate age-related alterations in the human hematopoietic system to have, in least in part, their origin already in the immature HSC level and find a number of genes differentially expressed within the HSCs compartment and with age.

Disclosures
No relevant conflicts of interest to declare.
From the Bone Marrow to the Thymic Niche
Authors: Susini Sandrine , Mouraud Séverine , Elkaim Elodie , Roullier Julien , Luce Sonia , Pellé Olivier , Bruneau Julie , Cavazzana Marina , Andre-Schmutz Isabelle 
Published: October 2019

Abstract
To generate T cells throughout adult life, the thymus must import hematopoietic progenitor cells from the bone marrow via the blood. The cellular and molecular mechanisms governing the circulation of thymus-seeding progenitor cells are well characterized in mice but not in humans. The aim of the present study was to characterize the molecular mechanisms and cellular components involved in thymus colonization by lymphoid progenitors (CD34+/CD10+/CD7-/CD24-) and the early steps of thymopoiesis under physiological conditions in humans. Our results demonstrate that circulating lymphoid progenitor cells express CCR9 and CXCR4 chemokine receptors, VLA-4, VLA-5 and VLA-6 integrins and PSGL-1 and CD44 adhesion molecules. We used in vitro migration and adhesion assays to validate the functional status of these markers.
As in the mouse, human circulating progenitor cells enter the thymus at the corticomedullary junction (CMJ). Once in the thymus, crosstalk with thymic epithelial cells causes the circulating progenitors to commit to the T-cell differentiation pathway. In order to characterize thymic niches and interactions between circulating progenitors and the thymic stroma, we undertook a chemokine/chemokine-receptor-focused gene expression analysis of sorted lymphoid progenitor cells and CMJ epithelial cells (based on the expression of EpCAM and Delta-like-4). We observed an unexpected gene expression profile for chemokines and chemokine regulators in thymus-seeding CD34+/CD10+/CD7-/CD24- cells and epithelial cells at the CMJ. The present results should help us to highlight candidate genes involved in the early steps of human thymopoiesis.

Disclosures
No relevant conflicts of interest to declare.
HDAC1 Can Deacetylate GATA-1 and Regulates Its Activity through a FOG-1 Independent Manner
Authors: Yan Bowen , Yang Tao , Huang Suming , Qiu Yi 
Published: October 2019

Abstract
The transcription factor GATA-1 is essential for erythroid and megakaryocytic cell differentiation and maturation. It has been well documented that GATA-1 can indirectly interact with histone deacetylase 1 (HDAC1) containing NuRD corepressor complexes through an association with FOG-1. Our previous work also shows that HDAC1 acetylation modulates the activity of FOG-1 associated NuRD complexes. Earlier studies show GATA-1 can be acetylated by p300/CBP, and the acetylation modulates GATA-1 binding activity to chromatin. However, it is not clear whether the acetylation can be reversed by a deacetylase. In this study, we found that GATA-1 can directly interact with HDAC1 in a FOG-1 independent manner. The interaction results in the deacetylation of GATA-1. We have identified two arginine sites within GATA-1 that are important for its interaction with HDAC1. The arginine to alanine mutation on these sites (2RA) can largely decrease the interaction of these two proteins, but doesn't affect its interaction with FOG-1, indicating that the direct interactions with HDAC1 and FOG-1 dependent association of NuRD complexes are separate events. The mutations also do not affect GATA-1 DNA binding activity in vitro. To further investigate the function of this interaction in erythropoiesis, we introduced wild type or the 2RA mutant of GATA-1 fused with estrogen receptor ligand binding domains into G1E cells, a GATA-1-null erythroid progenitor cell line. Upon estradiol induction, GATA-1 2RA inhibits the differentiation in G1E cells. We performed RNA sequencing to study the effect of GATA-1 2RA in gene expression in comparison with wild type GATA-1 and GATA-1-V205M (a mutation abolished binding with FOG-1) on a genome wild scale. GATA-1 2RA affects GATA-1 function in both GATA-1 activated and repressed genes. Although there is some overlap between 2RA and V205M mutations in both activated and repressed genes, more genes affected by these two mutations are different. Thus, this study unveils a novel regulation for GATA-1 by its direct interaction with HDAC1 during hematopoiesis.

Disclosures
No relevant conflicts of interest to declare.
Exosomes Secreted By Multiple Myeloma Cell Lines (MMCLs) and Bone Marrow Stromal Cells (BMSCs) Reciprocally Modulate Cell Adhesion and Impact on MM Cell Migration and Bortezomib-Induced Apoptosis
Authors: Udi Josefina , Voll Reinhard , Ureta Daniela , López Margarita , Wäsch Ralph , Álvarez Élida , Engelhardt Monika 
Published: October 2019

Abstract
Introduction: In MM pathogenesis, the interaction of plasma cells and the bone marrow microenvironment plays a crucial role. Exosomes are secreted by nearly all cell types, including cancer cells, and have been implicated in intercellular communication, drug resistance and tumor progression. We therefore purified, characterized and studied the influence of exosomes secreted by MMCLs and BMSCs on cell adhesion, bortezomib-induced apoptosis and cell migration.
Methods: Exosomes were purified from the supernatants of MMCLs (IM-9, MM.1S, U266, RPMI8226) and M2-10B4 BMSCs after 72h of culture with RPMI1640+10% exosome-depleted FBS by differential centrifugation. Purity and quality of exosomes was confirmed by electron microscopy and western blot for CD63. Exosomal protein concentration per 5x106 cells was quantified by Bradford. CD44 and ICAM1 were evaluated by flow cytometry. Apoptosis was assessed via annexin V/7-AAD-staining. Chemotaxis to M2-10B4 exosomes (100-500μg/ml) was studied using 24-well chemotaxis chambers. Migrated cells were counted by flow cytometry.
Results: Exosomes from IM-9 cells were identified by electron microscopy, thereby showing a characteristic saucer-like morphology, ranging in diameter between 30 and 120nm. Both MM cell and M2-10B4 exosomes expressed CD63. Mean exosomal protein concentration was 1.4μg/μl for M2-10B4 and varied for MMCLs between 1.2 and 1.7μg/μl. M2-10B4 cells expressed higher levels of both adhesion molecules CD44 and ICAM-1 when incubated with exosomes from MMCLs compared to basal conditions, however, not reaching statistical significance. Conversely, M2-10B4 exosomes induced a significant increase in CD44 expression on IM-9 cells (p=0.03), reflecting an increased adhesion to components of the extracellular matrix that might contribute to cell-adhesion mediated drug resistance. IM-9 apoptotic cells after 24h with 10nM bortezomib decreased from 72% to 61% in the presence of M2-10B4 exosomes and from 74% to 62% with IM-9 exosomes, suggesting a lower sensitivity to bortezomib. Migration of IM-9 cells slightly increased with 200μg/ml M2-10B4 exosomes and decreased with higher exosome concentrations.
Conclusions: Our findings underline the role of M2-10B4 exosomes on adhesion and migration of one representative MMCL (IM-9), and the impact of M2-10B4 and IM-9 exosomes on bortezomib cytotoxicity in this MMCL, suggesting their involvement in a paracrine/autocrine loop modulating treatment response and MM cell survival.

Disclosures
No relevant conflicts of interest to declare.
Preliminary Study on the Abnormal DNA Methylation in T Cells from the Patients with Immune Related Pancytopenia
Authors: Shao Zonghong , Ren Yue , Fu Rong 
Published: October 2019

Abstract
Objective To explore the global DNA methylation and the expression of regulatory genes for methylation in CD4 + T cells of the patients with immune related pancytopenia (IRP) and explore the role of methylation in pathogenesis of IRP.
Methods Thirty IRP patients (untreated, n=15; remission, n=15) and 15 healthy donors as controls were enrolled from December 2012 to December 2013. CD4+ T cells were sorted by immunomagnetic separation. The global DNA methylation was tested with enzyme-linked immunosorbent assay (ELISA). The mRNA levels of DNA methylation-related regulating genes, DNA methyltransferases (DNMTs) and methylated CpG binding proteins (MBDs), were measured by real-time quantitative polymerase chain reaction (RT-PCR).
Results The level of global DNA methylation in peripheral blood CD4+ T cells of untreated IRP patients (3.525%±2.046%)and remission patients (4.790%±1.471%) were significantly lower than that of normal controls (10.101%±3.449%) respectively (both P<0.05). DNMT3b mRNA level of untreated IRP patients (1.332±0.785) was significantly lower than that of normal controls (2.077±1.059) in CD4+T cells (P<0.05). The mRNA expression of MBD2 was significantly higher in CD4+ T cells from untreated and remission IRP patients (2.999±1.601, 2.055±1.576) than that in controls (0.581±0.247) (both P<0.05). The MBD4 mRNA level was significantly higher in CD4+ T cells from untreated IRP patients (2.736±2.719) compared to that in normal controls (1.167±1.006) (p<0.05). DNMT3b mRNA expression and CD4+ T cell DNA methylation to be positive correlated within IRP patients (r=0.569, p<0.01). The MBD2 mRNA expression negatively correlated with CD4+ T cell DNA methylation and the ratio of Th1/Th2 (r=-0.763, p<0.001; r = -0.652, p<0.05). The global methylation of CD4+ T cells negatively related to the ratio of CD5+ B cells (r= -0.439, p<0.05).
Conclusions The globe DNA hypomethylation and abnormal expression of DNA methylation-related enzymes in peripheral blood CD4+ T cells may be related with the pathogenesis of IRP.

Disclosures
No relevant conflicts of interest to declare.
Should We Hold the Point of allogeneic Hematopoietic Stem Cell Transplantation for Paroxysmal Nocturnal Haemoglobinuria in Eculizumab Era?
Authors: Chen Feng , Depei Wu , Sun Aining , Tang XiaoWen , Fu Chengcheng , Ma Xiao , Xue Shengli , Jin Zhengming 
Published: October 2019

Abstract
PNH is a rare haematological disorder due to the clonal expansion of abnormal haemotopoietic stem cells ,carrying a mutation in the PIG-A gene. Eculizumab ,as a humanised anti-complement component (C5), has proven highly effective in controlling intravascular haemolysis, and is expected to result in a remarkable improvement of survival. Should Eculizumab thus be offered as first treatment to all PNH patients? Allogeneic hematopoietic stem cell transplantation is still the only curative treatment for PNH by eradicating the abnormal PNH clone, leading to definitive survival improvement and QOL benefits.
Methods We assessed the long-term clinical and hematologic results in 13 PNH patients who received Allogeneic hematopoietic stem cell transplantation in our centre between Jan 2010 and Apr 2014. The patients were aged 13 to 54 (median 24 years). Eight donors were HLA -haploidentical and 5 were HLA-identical (3 siblings and 2 unrelated donors).
Results Eleven patients received a myeloablative conditioning consisting of busulfan and cyclophosphamide and 2 were given a reduced intensity conditioning consisting of fludarabine, busulfan and ALG (2 HLA-identical sibling donor). All patients achieved sustained engraftment with a median time of 11 days (range 11-26) to reach 0.5 ×109/L neutrophils and 15 days (range 12-126) to reach 20 ×109/L platelets. The overall cumulative incidence of grade II-IV acute GvHD was 15.4% and 2 patients showed limited chronic GvHD in evaluable 12 patients (cumulative incidence 16.7%).Transplant-related mortality for all patients was 15.4% (1 due to TMA and 1 due to severe pulmonary infection). As of Aug 1,2014, 11 patients are alive with complete hematologic recovery and no evidence of PNH following a median follow-up of 13 months. The 3-year Kaplan-Meier probability of disease-free survival was 80.2±12.8%. No thromboembolic event nor recurrence of the disease were reported following transplant.
Conclusions The findings of this study confirm that most patients with PNH may be definitively cured with allogeneic hematopoietic stem cell transplantation.

Disclosures
No relevant conflicts of interest to declare.
Defining the Cell Surface Landscape in B-Cell Malignancies: Comprehensive Profiling of Surface Antigens for Biomarker Discovery and Patient Stratification
Authors: O'Donnell Erika A , Alvendia Erica L , Wehrman Tom S , Krutzik Peter O 
Published: October 2019

Abstract
Flow cytometric analysis of surface marker expression is a fundamental component of diagnosis, classification, and monitoring for many hematologic malignancies. However, most clinical implementations of surface immunophenotyping focus on only a handful of markers. A comprehensive survey of CD markers and cell-surface receptors could provide a higher resolution and more accurate characterization of cancer lineages, as well as enabling new insights into the cellular changes resulting from disease progression or therapeutic intervention. To this end, we collated and evaluated antibodies to over 400 known markers and receptors. To establish the baseline and variability of each marker in major B cell subsets within normal human blood, a protocol was established to co-stain each sample for CD19, CD20, CD27, IgD, CD38, and CD24 simultaneously with each of the 400 markers. Antigen abundance and expression frequency were evaluated to establish a surface marker profile for each B-cell subset and to establish the variation inherent in each marker across the normal human population. To validate the application of this panel for cancer immunophenotyping, 20 malignant B-cell lines were selected for comprehensive surface characterization. Prior to staining, each cell line was barcoded with a combination of fluorescent dyes to generate a unique fluorescent signature. This approach allowed multiple cell lines to be combined into a single well for staining and analysis, enabling the rapid evaluation of hundreds of surface antigens using only 1-2 million cells from each line. Hierarchical clustering of data from cancer lines and normal B cell subsets allowed the identification of markers useful for discriminating between B-cell cancer subtypes, as well as markers that differed between normal and malignant B cells. These studies have resulted in the optimization of a protocol and antibody panel for use in profiling surface protein expression on human blood cells and cell lines. Application of this procedure to clinical samples may provide information useful for biomarker discovery, improved stratification of hematologic malignancies, and the identification of new drug targets.

Disclosures
O'Donnell: Primity Bio: Employment. Alvendia:Primity Bio: Employment. Wehrman:Primity Bio: Employment. Krutzik:Primity Bio: Employment.
Higher MMP9 and FGF2 Protein Levels and Elevated MMP9/TIMP Ratios Were Associated with JAK2V617F Mutation Regardless of Allele Burden in Myelofibrosis
Authors: Lima Luciene Terezina , Nunes Daniela Prudente Teixeira , Gomes Guilherme Wataru , Chauffaille Maria de Lourdes L. F. , Silva Maria Regina Regis , Ponce Cesar Cilento , Castro Fabiola Attié , Santos Marcos Tadeu , Guerra-Shinohara Elvira M 
Published: October 2019

Abstract
Background: Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) are involved in tumor neoangiogenesis which plays an important role in hematological neoplasias progression and prognosis. Myelofibrosis (MF) and essential thrombocythemia (ET) are myeloproliferative neoplasms (MPNs) resulting from acquired hematopoietic stem cell mutations. Most patients share the JAK2V617F mutation and may exhibit substantial overlap. The molecular mechanisms involved in MPNs pathogenesis are not completely elucidated. In this context, evaluation of genes and proteins related with angiogenesis could be useful to better understand MPN pathobiology.
Aims: To investigate the mRNA and protein expressions of MMPs and TIMPs and their relationship with plasma markers of angiogenesis (VEGFA and FGF2) and JAK2V617F mutation status in MF and ET patients.
Methods: Forty-three MPNs patients (MF=29 and ET=14) and 36 control subjects were studied. The controls were matched with patients according to age and gender. MPN diagnosis was defined according to the 2008 WHO criteria. The MMP2, MMP9, TIMP1 and TIMP2 mRNA expression from peripheral leukocytes and serum proteins levels were measured by real time PCR and Luminex technology. VEGFA and FGF2 concentration were measured in plasma from MPN patients and controls. JAK2V617F mutation status and allele burden and MPL W515K/L analysis were performed in total leukocytes DNA by real time PCR, using Taqman MGB probes.
Results: Higher MMP2, MMP9 and TIMP1 mRNA expression were observed in MF patients compared to controls (P<0.05). The frequencies of JAK2V617F mutated patients were 62.1% for MF and 35.7% for TE. The allele burden was similar in MF and ET groups. MF patients positive for JAK2V617F presented higher protein levels of MMP9 (N=18, P=0.008) and FGF2 (N=18, P=0.009) when compared with non-carriers of mutation (N=11). In contrast, no correlations were found between allele burden and MMP9 (r=0.153, P=0.558, N= 18) or allele burden and FGF2 (r=0.069, P=0.794, N=18). Carriers of JAK2V617F in MF group (N=17) showed elevated MMP-9/TIMP1 and MMP9/TIMP2 ratios of protein levels than those MF JAK2V617F negative (N=11, P<0.025). ET patients JAK2V617F positive presented higher MMP9 mRNA level (N=5) compared with negative ones (N=9, P=0.009), but no difference was observed in protein levels. None of patients presented MPL W515K/L mutation.
Conclusions:Our findings suggest that MF patients with JAK2V617F mutation exhibit higher MMP9 and FGF2 protein levels and elevated MMP9/TIMP ratios, which are not associated with JAK2V617F allele burden.
Financing: FAPESP 2012/12957-5

Disclosures
No relevant conflicts of interest to declare.
Effect of Hes1 on AML Cells: A Potential Therapeutic Target
Authors: Tian Chen , Jia Yongsheng , Hu Dongzhi , Zhang Yizhuo 
Published: October 2019

Abstract
Notch signal pathway is an important mediator of growth and survival in several cancer types, while Notch pathway genes function as oncogenes or tumor suppressors in different cancers. Although aberrant Notch activation contributes to leukemogenesis in T cells, the role of Notch pathway in acute myeloid leukemia (AML) remains unclear. To address this problem, we investigated the expression levels of its downstream effector Hes1 and p21 in primary AML cells and cell lines by realtime PCR and western, and found that both of them was weak in these cells. Induced activation of Hes1 consisently led to AML growth arrest and apoptosis, which was associated with enhanced p21 expression. Besides, overexpression of Hes1 inhibited growth of AML cells in vivo. In conclusion, we reported that Hes1 mediated growth arrest and apoptosis of human AML cells, and demonstrated a novel tumor suppressor role for Hes1 in AML.

Disclosures
No relevant conflicts of interest to declare.