bluuebunny/crossref_metadata_2025_split huggingface dataset:
HIF-2alpha Associated Familial Erythrocytosis Supports the PHD2-HIF- 2alpha-VHL Axis as the Major Regulator of Erythropoietin Production
Authors: Percy Melanie , Furlow Paul W , Beer Philip A , Campbell Gavin , Dekker Adriaan W. , Green Anthony R. , Li Xiping , Sutherland Scott , Oscier David , Rainey M. Glenn , Rivera Enid , van Wijk Richard , Wood Marion , Lappin Terence R.J. , McMullin Mary Frances , Lee Frank S 
Published: April 2020

Abstract
Transcriptionally erythropoietin (Epo) synthesis is tightly regulated by the hypoxia inducible factor (HIF), which is composed of one alpha and one beta subunit that are constitutively expressed. The beta subunit is non-variable, but three different alpha subunits give rise to three isoforms of HIF. The alpha subunit is proteasomally regulated in the presence of oxygen by hydroxylation of the proline in the LXXLAP motif of the oxygen dependent degradation (ODD) domain of HIFalpha, catalysed by members of the prolyl hydroxylase domain (PHD) family of enzymes. This allows the von Hippel Lindau (VHL) protein to associate with the alpha subunit, which is subsequently tagged with ubiquitin and degraded by the proteasome. Any defect in the oxygen sensing pathway that allows the alpha subunit to escape proteasomal regulation leads to elevated expression of HIF target genes.
Recently mutations in both VHL and PHD2 have been identified in a cohort of patients with erythrocytosis, but no mutations were found in the ODD domain of HIF1alpha. Instead, investigation of the homologous region in HIF-2alpha revealed four different mutations, Pro534Leu, Met535Val, Gly537Arg and Gly537Trp in seven individuals/families. Affected individuals presented at a young age with elevated serum Epo. Several individuals have a clinical history of thrombosis, but no evidence of a von Hippel Lindau-like syndrome.
To define how the four mutations relate to the erythrocytosis phenotype functional assays were performed in vitro. Binding of PHD2 to the four HIF-2alpha mutants was impaired to varying degrees, with both the Gly537 mutants showing the greatest reduction. The association of VHL with the hydroxylated Met535Val mutant peptide was similar to wild type HIF- 2alpha, but was decreased in the other three HIF-2alpha mutants. Expression of three HIF- 2alpha target genes, adrenomedullin, NDRG1 and VEGF, was significantly up-regulated in cells stably transfected with the mutants under normoxia compared to wild type HIF-2alpha. Mutations in the ODD domain of HIF-2alpha disrupt proteasomal regulation by reducing the association with PHD2 and hence hydroxylation. Furthermore the binding of VHL is also impaired, even when HIF-2alpha is hydroxylated. Examination of the three-dimensional structure of hydroxylated HIF-1alpha bound to VHL confirms that amino acids close to site of hydroxylation (Pro-531 in isoform 2) are important for this association. These observations, together with recent studies utilising murine models of erythrocytosis, support the PHD2-HIF-2alpha-VHL axis as the major regulator of erythropoietin.
Smoking and Hodgkin Lymphoma: A Meta-Analysis
Authors: Pascual Sheila , Milani Cannon , Mitri Joanna , Castillo Jorge 
Published: April 2020

Abstract
INTRODUCTION: The etiology of Hodgkin lymphoma (HL) is largely unknown. However, certain associations have been noted, such as familial factors and infection with viruses. Smoking has been associated with the development of multiple malignancies and some studies have reported an association between HL and smoking but the relationship of tobacco use with lymphomas in largely unclear.
OBJECTIVE: To investigate the potential relationship between tobacco use and the development of HL using a meta-analysis methodology of retrospective, case-control studies.
METHODS: An extensive search was conducted using Pubmed/MEDLINE through July 2008. Case-control studies that reported odds ratio (OR) and 95% confidence intervals (CI) or allow for those values to be calculated were included in our analysis. Case reports, editorials, letters to the editor, review articles and prospective studies were excluded. The smoking status was then subdivided in three groups: never smokers, former smokers and current smokers. Meta-analyses were performed comparing the risks of former and current smokers against the risk of never smokers of developing HL. Fixed and random effects models were used to assess for heterogeneity.
RESULTS: Seven case-controls studies, accounting for 3201 cases and 15268 controls were included in the analysis. Most of the articles reported OR adjusted for age, sex and educational level. In former smokers, the OR was 0.73 (95% CI, 0.65 – 0.82) when compared to never smokers; no heterogeneity was detected. The current smoker group had an OR of 1.70 (95% CI, 1.36 – 2.13) when compared to the never smoker group; some heterogeneity was detected in this group (p &lt; 0.003, I2 = 69.6%).
CONCLUSIONS: Despite the heterogeneity observed in the analysis, the current smoker group seems to have a 70% increased risk of developing HL. Although a cause-effect linkage between tobacco use and HL is difficult to prove, further basic and translational research is necessary to clarify the potential etiological role of smoking in HL.
FLT3 Internal Tandem Duplication Is Associated with High Relapse Rate and Central Nervous System Involvement in Acute Promyelocytic Leukemia
Authors: Tashiro Haruko , Oka Yoko , Sugao Toshihiko , Shirasaki Ryosuke , Noguchi-Mizutani Mitsuho , Akiyama Nobu , Kawasugi Kazuo , Shirafuji Naoki 
Published: April 2020

Abstract
The internal tandem duplication of FMS-like tyrosine kinase 3 (FLT3/ITD) is now accounted as one of important risk factors for the prognosis in acute myelogenous leukemia (AML) patients with normal karyotype. In all-trans retinoic acid era, acute promyelocytic leukemia (APL) has become the most curative subtype of adult AML, and usually hematopoietic stem cell transplantation is not selected for APL patients in 1st complete remission (CR) or 2nd CR without minimal residual disease (MRD) at the polymerase chain-reaction level; however, relapses are still reported to occur in 15–25% of cases after achieving 1st CR without MRD, and some risk factors are proposed including the difference of breakpoint region of the translocation, and leukocyte count at the onset. The incidence of FLT3/ITD in APL has been reported 17–35%; however, it is still a matter of debate that the presence of FLT3/ITD is associated with poor prognosis in APL patients. Thus, we focused on the incidence, clinical features, and prognostic implications of FLT3/ITD in adult newly diagnosed 17 APL patients (median age of 45 years, range 22–67 years, 11 males and 6 females, and median total leukocyte count (TLC) of 2.0 × 109/L, range 0.4– 156.1 × 109/L). All patients achieved CR. The incidence of the presence of FLT3/ITD was 29.4%. The median TLC was significantly higher in FLT3/ITD positive group (median TLC of 9.4 × 109/L) as compared in the negative group (median TLC of 1.2 × 109/L). Higher LDH level in blood was also observed in FLT3/ITD positive group. The rate of central nerves system (CNS) involvement was significantly higher in FLT3/ITD positive group than that in the negative group (60 % vs. 0%). Relapse rate was also significantly higher in FLT3/ITD group (100 % vs. 0%). These data indicate that the presence of FLT3/ITD is associated with the high relapse rate and CNS involvement in APL patients, and implies one of poor prognostic factors.
Identification of a Homeobox-Like Protein Immune Response in Bone Marrow Transplant Patients
Authors: Arden Nilou , Chen George L. , Miklos David 
Published: April 2020

Abstract
The identification of novel immune targets following hematopoietic cell transplantation (HCT) promises to identify clinically important graft-v-leukemia and graft-v-host disease targets. High-density protein microarrays present thousands of human proteins and facilitate broad-band multiplexed quantitative auto and alloreactive antibody screening. Our studies utilized both male and female patient plasma which were collected pre-transplant and one year post-transplant to identify new immune response targets. The tetra-peptide repeat homeobox-like protein (TPRXL) was initially identified by protein microarray as a potential biomarker in one MDS and four AML transplant patients. We developed an ELISA assay by affinity purifying TPRXL expressed in E.coli, and tested plasma samples collected from 80 HCT patients 1–2 years after HCT. A 42% sero-positive incidence was found across the group of post transplant samples (p-value &lt;0.001). Located at 3p25.1 of the human genome, TPRXL encodes a 257 amino acid and 25 kDa protein containing a significant percentage of serine tetra-peptide repeats. Being a member of the TPRX homeobox gene family, the protein’s secondary structure and function lead to direct interaction with DNA and tight regulation of gene expression. Presently, of the 18 reported TPRXL transcript variants a majority are nearly identical and are found to be highly expressed in early embryonic development, in the placenta, oral tissues and the ascites. Results indicate disease specificity with prevalence of post transplant response in patients with Mantle cell lymphoma (MCL). In addition, patients with chronic GVHD of the oral pharyngeal tissues are predominant in our analysis. In silico studies show that TPRXL is indeed differentially expressed in acute pro-myelocytic leukemia (PML) and acute lymphocytic lymphoma (ALL) cell lines as compared to controls. Since bioinformatics expression data available for MCL cell lines are scarce, perhaps due to fewer reports or studies in this area, it is clear that our novel findings will contribute to currently available information for this disease type. In sum, protein microarray technology represents a powerful tool in facilitating the identification of novel disease associated antigenic targets and that TPRXL is potentially one such prognostic marker.
Study on the Dendritic Cell Subsets and Their Relationship with the Expressions of T-Bet and GATA-3 in Peripheral Lymphocytes of Severe Aplastic Anemia Patients
Authors: Shao Zonghong , Jun Wang , Fu Rong , Li Lijuan , Liu Hui , Qu Wen , Rua Erbao , Liang Yong , Liu Hong , Wu Yuhong , Wang Guojin , Song Jia , Wang Huaquan , Xing Limin , Guan Jing , Wang Xiaoming 
Published: April 2020

Abstract
Objective To investigate the effects of the dendritic cell (DC) subsets and the transcriptive factors, T-bet and GATA-3, on immune imbalance in the pathogenesis of acquired severe aplastic anemia (SAA).
Methods The mDCs(HLA-DR+Lin−CD11c+) and pDCs(HLA-DR+Lin−CD123+) in peripheral blood mononuclear cells (PBMNC) were measured with flow cytometry (FCM), the expressions of T-bet mRNA and GATA-3 mRNA in PBMNC with semiquantitive RT-PCR and the plasma levels of IFNγ and IL-4 with ELISA in 29 SAA patients (21 untreated and 8 recovered) and 16 healthy controls.
Results The percentages of mDCs in PBMNC were (0.44±0.24) % and (0.73±0.30) % in untreated and recovered SAA patients respectively, either of which was higher than that of controls (0.29±0.10%) (P&lt;0.05). The percentage of pDCs in the untreated cases was lower than that of the recovered or controls [(0.18±0.14) % vs. (0.28±0.20) % and (0.29±0.13) %] (P&lt;0.05). mDCs/pDCs ratios were 3.45±2.71 and 2.90±0.95 in untreated and recovered groups respectively, either of which was higher than that of controls (1.15±0.56) (P&lt;0.05). No statistical difference of mDCs/pDCs was found between untreated and recovered patients (P&lt;0.05). The relative mRNA expressions of transcriptive factor T-bet were 0.37±0.07, 0.20±0.07 and 0.17±0.05 in 3 groups, respectively. The expression of T-bet of untreated patients was higher than that of recovered patients or healthy controls (P&lt;0.05). There was no statistical difference of GATA-3 expression among 3 groups(P&gt;0.05). T-bet/GATA-3 ratio was 0.72±0.13 in untreated patients, which was higher than that of recovered patients (0.33±0.08) or controls (0.35±0.11). The plasma level of IFNγ in the untreated cases was (50.9±1.1)ng/L, which was higher than that of recovered [(49.7±0.9) ng/L] or controls[(49.7±0.7) ng/L]. There were significant positive correlation between T-bet and mDCs/pDCs (r=0.445, P&lt;0.01), as well as T-bet and IFNγ (r=0.402, P&lt;0.01).
Conclusion Both mDCs/pDCs and T-bet/GATA-3 ratios in SAA were higher and positively correlated which might contribute to the cascade activation of T lymphocytes. Immunosuppressive therapy for SAA should be kept on until both ratios become normal.
Performance Comparison of Commercial Fixing and Permeabilizing Reagents
Authors: Joe Edward , Frey Tom 
Published: April 2020

Abstract
Flow cytometry is commonly used to characterize cellular antigens expressed both on the cell surface and inside the cell. As the technology advances, more researchers are using this tool to study intracellular activity to understand how cells communicate and respond to their environment. Intracellular targets of interest include cytoplasmic antigens and proteins, nuclear antigens and proteins, cytokines, etc. The quality of intracellular staining depends on the ability of the permeabilizing reagent to permeabilize the cell membrane for antibodies to get to the target of interest without destroying the cell. Five different commercial permeabilizing reagents were evaluated in a small study, Caltag Fix and Perm® Kit, BD IntraSure™ Kit*, BD FACS™ Permeabilizing Solution 2 (FACS Perm 2), 1X FACS Lysing Solution, and BD Cytofix/Cytoperm™ Kit. Five categories of performance were evaluated: cell recovery, maintenance of surface staining, preservation of light scattering properties, intracellular staining, and background staining. Cell recovery was assessed using CD45 APC and BD TruCOUNT™ tube; surface staining maintenance was assessed using CD45 APC, CD45 PerCP-Cy5.5, CD19 PE, and CD20 PerCP; intracellular and background staining were assessed using anti-bcl-2 FITC, anti-Zap 70 PE, anti-TdT FITC, anti-Ki-67 FITC, and CD3 FITC/anti-MPO PE/CD79a PerCP-Cy5.5 reagents. Caltag and BD IntraSure Kits performed well in the category of cell recovery. Caltag Kit, BD IntraSure Kit, and FACS Lysing Solution maintained cell surface staining after cell permeabilization. BD IntraSure Kit and FACS Lysing Solution preserved light scattering properties of cells after permeabilization. Using signal-to-noise ratio as the criterion to evaluate background and intracellular positive staining, FACS Lysing Solution and FACS Perm 2 detected anti-bcl-2 FITC; BD IntraSure Kit and FACS Lysing Solution detected anti-TdT FITC; BD IntraSure Kit, FACS Lysing Solution, FACS Perm 2, and BD Cytofix/Cytoperm Kit detected anti-Ki-67 FITC; BD IntraSure Kit, FACS Lysing Solution, and FACS Perm 2 detected anti-Zap 70 PE; all kits detected CD3 FITC/anti-MPO PE/CD79a PerCP-Cy5.5. Results showed that there is no universal permeabilizing reagent for all intracellular staining assays. Each test reagent has its own strength and weakness. Choosing the best permeabilizing reagent for a particular assay depends on the target of interest and the output of the assay.
Does PDG-PET Add Additional Prognostic Information to CT Staging in Lymphoma Patients?
Authors: Sanchez-Gonzalez Blanca , Trampal Carlos , Solano Alberto , Pedro Carmen , Maiques Jm , Alvarez Alberto , Gimeno Eva , Abella Eugenia , Besses Carles , Salar Antonio 
Published: April 2020

Abstract
BACKGROUND: 18-Fluoro-deoxyglucose positron emission tomography (FDG-PET) is a noninvasive imaging tool for initial staging and evaluation of treatment response in Hodgkin’s lymphoma (HL) and non-Hodgkin’s lymphoma (NHL). PET imaging can provide additional information to conventional imaging as computer tomography (CT).
AIM: The aim of this study was to evaluate the role of PET in improving the staging and if this fact could have an impact in prognosis, in compared to CT.
METHODS: From February 2005 to January 2008, one hundred ten consecutive patients with NHL (49 aggressive, 32 indolent and 3 unclassifficated) and HL (26 patients) were staged according to the Ann Arbor classification with CT and bone marrow biopsy (BMb). As well, all patients underwent to FDG-PET body before treatment. International Prognostic Index (IPI) was calculated following standard staging (IPI-st) and with standard staging plus FDG-PET (IPI-PET) in NHL.
RESULTS: CT and FDG-PET were concordant in 84 patients (66%). In 37 patients (34%) discordance was observed: in 7 patients FDG-PET was negative, in 11 patients FDG-PET detected more bone lesions, in 11 patients FDGPET uptake in other lymph nodes, including spleen infiltration, and in 8 patients FDGPET detected less lymph nodes or organ involvement. However, additional information of FDG-PET changed in staging in 21 out of 37 patients with discordant findings but the IPI risk changed in only 5 patients. Discrepancies CT and FDG-PET were seen in similar proportion in all lymphoma subtypes. Overall survival curves at two years showed no differences between IPI-st and IPI-PET (IPI-st 0–1: 97%, 2–3: 89%, 4–5: 48%; IPI-PET 0–1: 96%, 2–3: 89%, 4–5: 48% ).
CONCLUSIONS: FDG-PET can provide additional information to CT in both aggressive or indolent non-Hodgkin lymphoma and Hodgkin lymphoma. CT and FDG-PET were discordant in 30 % of the patients but only half of them showed change of stage. IPI risk was modified in few patients with NHL. Inclusion of patients is ongoing.
Alternate Splicing Is a Frequent Event and Impacts Clinical Outcome in Myeloma: A High-Density Exon Array Analysis of Uniformly Treated Newly-Diagnosed Myeloma Patients
Authors: Munshi Nikhil C. , Li Cheng , Minvielle Stephane , Amin Samir B , Moreau Philippe , Magrangeas Florence , Anderson Kenneth C , Avet-Loiseau Herve 
Published: April 2020

Abstract
Alternate splicing is an important post translational change that alters specificity of gene function. Misregulation of alternative splicing has been implicated in number of disease processes including cancer. We have here analyzed alternate splicing in myeloma using high throughput exon array analysis. The GeneChip Human Exon 1.0 ST Arrays used in this investigation not only provides information on expression levels for genes, but as the probe sets are spread evenly over each exon, the array data also provides information on presence of each exon and identify recurrent alternately spliced genes. We conducted a study in series of 170 newly-diagnosed patients with multiple myeloma treated homogeneously in tandem transplantation IFM trials. RNA isolated from CD138 purified MM cells collected at the time of diagnosis were analyzed using the GeneChip Human Exon 1.0 ST Arrays. The dChip software was modified to analyze exon array data. We first normalized gene-level expression values across samples, and then identified differentially expressed exons between two sample groups. These exons are candidates for alternatively splicing events. We observed over 100 genes with candidate alternate splicing events, which have up or down-regulation of an exon relative to the baseline group in more than 20% patients. Dividing the group based on response, 52 alternately spliced exons were identified as influencing response to therapy. The genes and their exons with highest frequency of alternate splicing were CD79A exon 5 and 6, EDNRB exon 2, RASSF1 exon 8, CD1d exon9, TGFBetaRII exon 1, Calmin exon2, CEACAM1 exon 7, MADH1 exon 7, TBX5 exon 2, Amyloid beta exon 14, Nit protein 2 exon 10, Thymidylate synthetase exon 7. Amongst these genes, 32 genes had the most influence on response with over 50% differential frequency in patients achieving CR. Similarly we have identified 85 alternately spliced genes as influencing overall survival between groups divided by less than or more than 48 month survival. The genes with highest frequency of alternate splicing were NOTCH2 exon 18, CXCL3 exon 8, CD9 exon 3 Calmin exon 2, TIMELESS exon 12, SPOUTY homolog 1 exon 4, Amyloid beta exon 14, Nit protein 2 exon 10, Cyclin A2 exon7, lymphocyte antigen 6 exon 7. Forty-nine genes within this group had the most influence on overall survival. Number of spliced variants were shared between both response and survival groups suggesting their biological and functional significance. Further validation of these alternate splicing is under investigations. This study highlights significant frequency of alternate splicing and points to the need for evaluation of not only the expression level of genes but also post translational modifications. The genes identified here are important target for therapy as well as possible immune modulation.
Clinical Efficacy of ATRA for Adult T-Cell Leukemia
Authors: Maeda Yasuhiro , Yamaguchi Terufumi , Hirase Chikara , Kanamaru Akihisa 
Published: April 2020

Abstract
We previously reported that all-trans retinoic acid (ATRA) inhibited growth in HTLV-I- positive T-cell lines and fresh cells from patients with adult T-cell leukemia. We here confirmed the clinical effects of ATRA in 20 patients with ATL. Twenty patients (n=20) with median age of 56 years (range 35–68 years) diagnosed with ATL received ATRA orally. ATRA was administered for a median of 25.7 days (range 14–56 days). Efficacy was described below; no CR case, PR case was 55%, NR case was 45%. In 7 acute cases, PR case was 4 (20%) and NR case was 3 (15%). In 3 lymphoma cases, no NR case and 3 PR cases (15%) was found. In 4 chronic cases, PR case was 1 (4%) and NR case was 3 (15%). In 6 skin type. PR case was 3 (15 %) and NR case was 3 (15%). Major side effects were headache (n=5), transient liver dysfunction (n=2), hyperlipidemia (n=2) and anorexia (n=1). No major toxicity was observed. These results indicated that ATRA might be a useful agent for skin involvement of ATL with safety.