bluuebunny/crossref_metadata_2025_split huggingface dataset:
Significance of Factor VIII Inhibitors by ELISA in Patients with Hemophilia A.
Authors: Brewer Jamie R , Harris Sandra , Green David , Zakarija Anaadriana 
Published: October 2019

Abstract
Background: The Bethesda assay traditionally has been used to detect Factor VIII inhibitors in patients with Hemophilia A, but recent evidence suggests that it is not sensitive to all inhibitors, particularly non-inhibitory or low-titer antibodies.
Methods: Patients with Hemophilia A without prior history of inhibitor were recruited. Study questionnaire collected demographic and clinical information, bleeding history and factor usage over the preceding 6 months. Functional status was assessed by the Hemophilia Activities List (HAL). Factor VIII inhibitor was assessed by both the Bethesda assay and Factor VIII inhibitor ELISA (GTI Diagnostics). T-test was performed to assess statistical significance.
Results: Data is available for 26 patients, 19 severe, 2 moderate and 7 mild. All subjects had a negative Bethesda assay, but 10 (39%) had a detectable inhibitor by ELISA. 9/10 inhibitor patients had severe hemophilia, while one had mild hemophilia. In severe hemophiliacs, there were no differences in age, HIV status, CD4 count, Hepatitis C positivity or viral load between those with and those without inhibitors. Inhibitors were more frequent in those using plasma-derived concentrates 4/5 (80%), than in those using recombinant products 6/14 (43%), p=0.15. There was no difference in bleeding frequency or functional status in patients with or without inhibitors, although those with inhibitors had more frequent infusions.(Table). In patients on prophylaxis, those with inhibitors had a higher bleeding frequency compared to those without an inhibitor, (p =0.2). 11 patients were not on prophylaxis and had a higher bleeding frequency (p = 0.02) than patients on prophylaxis irrespective of inhibitor presence. However those with inhibitors required more factor doses per bleed compared to those without an inhibitor (4.4 vs. 1.5, p=0.16) even though the mean factor dose was the same (25.3 units/kg vs 25.2 units/kg).
Conclusions: The Factor VIII ELISA assay detected inhibitors in 39 % of Hemophilia A patients who had undetectable inhibitors by standard Bethesda assay. This data suggests that these inhibitors may be clinically relevant, given that inhibitor patients who are not on prophylaxis require more doses of factor per bleeding event. Further study is necessary to determine mechanism and clinical significance of these Factor VIII inhibitors.
Table. Characteristics of severe hemophilia patients with and without ELISA Factor VIII inhibitor
All severe (n=19) Inhibitor (n=9) No inhibitor (n=10) Age 43.4 40.8 45.7 Plasma-derived factor 5 (26.3%) 4 (44.4%) 1 (10%) Total bleeds/6 months 8.6 8.1 9.0 Muscle bleeds/6 months 1.3 0.8 1.7 Joint bleeds/6 months 7.3 7.3 7.3 Factor infusions/6 months 50.9 57.2 45.2 On prophylaxis 8 (42%) 4 (44.4%) 4 (40%) Total bleeds/6 months 4.3 5.8 2.8 Not on prophylaxis 11 (58%) 5 (55.6%) 6 (60%) Total bleeds/6 months 11.7 10 13.2 Factor doses/bleed 2.9 4.4 1.5
Health-Related Lifestyle among Adult & Pediatric Patients with Idiopathic Thrombocytopenic Purpura in the United Kingdom.
Authors: Sarpatwari Ameet , Watson Shirley , Anderson Howard , Provan Drew , Newland Adrian 
Published: October 2019

Abstract
Idiopathic thrombocytopenic purpura (ITP) is an autoimmune condition characterized by autoantibody-mediated platelet destruction and suboptimal megakaryocytic production. Primarily acute (< 6 months) in duration among children, ITP manifests predominantly chronically among adults, increasing susceptibility to bleeding events.
Despite recent growth, published literature on health-related quality of life in ITP remains limited. The objective of our investigation was to identify lifestyle concerns associated with ITP among adult and pediatric patients in the United Kingdom.
In collaboration with ITP specialists, a 43 question, closed-field lifestyle survey was developed, addressing social engagement, work and school performance, sports and activities, treatment, and travel. Patient members of the United Kingdom ITP Support Association (N = 1,767) were asked to complete and return mailed surveys. Pearson’s chi-square and Fisher’s exact tests were used to evaluate differences in dichotomized variables between groups.
790 (45%) completed surveys were returned. As illustrated in the table below, roughly one-quarter of adults (≥ 16 years) and one-fifth of children reported ‘always’, ‘sometimes’, or ‘often’ missing school or work owing to fatigue and having encountered difficulty obtaining insurance. Nearly one-third of adults further revealed having an elective surgery delayed owing to a low platelet count.
Disparities were noted across gender with regard to bruise concealment (adults: p < 0.01; children: p = 0.03) and suspicions of subjection to violence (adults: p < 0.01; children: p = 0.49). In contrast with adults, pediatric patients reported being more likely to having requests for referral denied (p = 0.03).
This study represents the largest quality of life investigation of ITP to date. Although the survey utilized remains to be validated, its findings nonetheless successfully highlight avenues for future investigation.
Subgroups: ‘Yes’ % Adults Children Question Total ‘Yes’ % N = 790 Male N = 199 Female N = 497 Male N = 51 Female N = 43 [1] Total number of responses recorded for the given question. Have you ever been unable to go to work or school because of tiredness and fatigue? 29%   710[1] 26%  186 31%  435 20%  49 30%  40 Have you had difficulty obtaining or been refused travel and life insurance? 29%  611 30%  540 18%  71 Have you ever had surgery (other than splenectomy for ITP) postponed or delayed because of a low platelet count? 30%  620 34%  158 30%  391 13%  39 19%  32 Do you try to hide your bruises? 29%  756 10%  190 37%  475 20%  50 42%  41 Are people ever suspicious that the bruises are a result of physical violence? 17%  750 5%  193 19%  468 31%  49 38%  40 Have you ever been refused a referral to an ITP specialist or hospital of 5%  574 3%  144 4%  358 10%  41 10%  31
Sensitive Detection of Survival of Paroxysmal Nocturnal Hemoglobinuria (PNH) Clones in Red Blood Cells, Granulocytes and Monocytes by High Resolution Multiparameter Flow Cytometry.
Authors: Movalia Mayur K , Illingworth Andrea 
Published: October 2019

Abstract
Paroxysmal Nocturnal Hemoglobinuria (PNH) is an acquired clonal stem cell disorder characterized by intravascular hemolysis due to GPI-deficient red blood cells sensitive to complement-mediated lysis. Accurate and sensitive detection of PNH-type cells has become important not only to diagnose PNH but also because studies have shown PNH-type cells may indicate favorable response to therapy and favorable prognosis in patients with aplastic anemia and myelodysplastic syndrome. Previous studies have suggested optimal testing for PNH-type cells by flow cytometry should be limited to within 48 hours after collection of whole blood. Our laboratory has developed a very sensitive and specific high resolution flow cytometric method for detecting PNH-type cells based on testing over 3,000 patients with known PNH, aplastic anemia, myelodysplastic syndromes and other bone marrow failure syndromes. The aim for this study was to determine the longevity of PNH clones in whole blood samples, the day-to-day variability of these clones and the rate of deterioration of the PNH clones compared to normal blood cells. We analyzed 10 whole blood samples from patients known to have PNH-type cells on seven consecutive days utilizing a two-color assay with GPA-CD59 for the red blood cells, a 5-color assay with FLAER-CD24-CD14-CD15-CD45 for the granulocytes and a 5 color assay with FLAER-CD33-CD14-CD64-CD45 for the monocytes. The results are summarized in the table below. The initial PNH clone sizes ranged from 0.02% to 90.8%. The PNH cells showed an overall similar level of deterioration to the normal blood cells with even minor PNH clones of 0.02% able to be detected at day 7. The day-to-day variability of PNH clone sizes was generally less than 10%, with smaller clone sizes showing a higher degree of variation, up to 20%, due to their smaller absolute numbers. Interestingly, Type III PNH red blood cells showed slightly better overall survival than normal red blood cells and were detected in modestly increasing percentages throughout the study. Based on this data, we propose that accurate detection of PNH type cells can be achieved up to seven days after collection of whole blood when utilizing high resolution flow cytometry. PNH Clone Size on Sequential Days as Percentage of Original PNH Clone Size
Original PNH Clone Sizes PNH Clone Sizes as Percentage of Original PNH Clone Size Cell Type Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Type III RBCs .02%–58.6% 102% 106% 107% 104% 108% 103% Granulocytes .29%–90.8% 100% 100% 93% 89% 79% 86% Monocytes .52%–89.9% 96% 96% 92% 94% 97% 85%
GATA-1 Enhances the Transcription of PIGM and Is Responsible for Increased GPI Expression in Erythrocytes in Inherited Glycosylphosphatidylinositol Deficiency: A Model for Tissue-Specific Housekeeping Gene Transcription Regulation
Authors: Costa Joana S , Nolasco Madalena , Almeida Antonio M , Roberts Irene , Layton Mark , Karadimitris Anastasios 
Published: October 2019

Abstract
Inherited glycosylphophatidylinositol (GPI) deficiency (IGD), characterized by thrombosis and epilepsy but minimal hemolysis, is caused by a c>g mutation at position −270 that disrupts binding of the transcription factor (TF) Sp1 to the core promoter of the mannosyltransferase-encoding gene PIGM. In IGD, the level of GPI expression varies considerably between hematopoietic cells of different lineages; for example, while in erythrocytes it is near normal, accounting thus for the absence of significant intravascular hemolysis, GPI synthesis in granulocytes is severely deficient suggesting erythroid-specific transcriptional regulation of PIGM. We tested the hypothesis that the erythroid-megakaryocytic-specific TF GATA-1 is critical regulator of PIGM transcription and it can overcome the transcriptional block imposed by the c>g mutation. Consistent with this hypothesis, we found that in luciferase assays a 2kb PIGM promoter construct had at least 3-fold higher activity when transfected in erythroid (K562) rather than myeloid (HL60), B lymphoid or epithelial (HeLA) cell lines. Co-transfection of the 2kb WT or c>g mutated construct with GATA-1 resulted in proportional increase of the transcriptional activity which was completely abolished when shorter, 0.6kb WT or c>g constructs were used, implying the presence of GATA-1 binding elements (BE) between −0.6 and −2kb. The functional significance of 3 such GATA-1 BE initially identified by bioinformatic analysis was investigated next. In vitro binding to each of these BE was shown in electromobility shift assays using appropriate oligos, nuclear extracts from K562 cells and anti-GATA-1 antibody while in vivo binding was demonstrated by chromatin immune-precipitation assays. The relative transcriptional activity of the 3 GATA-1 BE was tested in luciferase assays by co-transfecting into HeLA cells WT or c>g 2 kb constructs with single, double or triple BE disrupted by site-directed mutagenesis. Overall the results suggested that the proximal of the 3 GATA-1 BE is the most active and that the transcription-enhancing effect of GATA-1 is diminished in the presence of the disease causing c>g mutation. Thus, as well as explaining the near normal synthesis of GPI in erythrocytes in IGD, our results also provide an important paradigm of co-operative function of a generic (Sp1) and tissue-specific (GATA-1) TF in the transcriptional regulation of a house-keeping gene such as PIGM.
Incidence of SCN-Assoicated Gene Mutations in Severe Congenital Neutropenia Patients in North America
Authors: Xia Jun , Bolyard Audrey Anna , Rodger Elin , Stein Steven , Aprikyan Andrew AG , Dale David C. , Link Daniel C. 
Published: October 2019

Abstract
Severe congenital neutropenia is a genetically heterogeneous syndrome associated with mutations in several different genes including ELA2, HAX1, GFI1, WAS, and CSF3R. The goal of this study was to define the mutation frequency of these genes in the North American SCN patient population. We also sequenced SBDS, since mutations of SBDS have been associated with congenital and acquired neutropenia. A total of 159 patients were identified in the North American Severe Chronic Neutropenia International Registry (SCNIR) for whom informed consent and genomic DNA samples adequate for sequencing were available. To accommodate our semi-automated high-throughput sequencing pipeline, 94 samples were chosen for sequencing. Since ELA2 sequencing had already been performed in most cases, preference was given to those samples without known ELA2 mutation. Among the samples, 73 were from patients with SCN, 4 with cyclic neutropenia, 10 with idiopathic neutropenia, 2 with Shwachman-Diamond Syndrome (SDS), and 3 with Barth syndrome. Two samples were excluded because of poor sequence quality. Singleton cases with validated mutations of GFI1 (N382S) and WAS (L270P) were observed. The N382S GFI1 mutation was associated with striking monocytosis. A novel nonsense mutation of GFI1 (R412X) was detected in one additional case. As expected, compound heterozygous mutations of SBDS were present in the two cases of SDS. In addition, heterozygous mutations of SBDS (84Cfs3X and Q94X) were observed in two cases of SCN. Typical truncation mutations of CSF3R were detected in 4 cases, all developing MDS or AML. Surprisingly, no mutations of HAX1 were detected. Considering only patients with a diagnosis of SCN who were from North America (125 of the total 159 cases), the incidence of ELA2 mutations was 68%. Eleven novel ELA2 mutations were identified. In 28.8% of cases, no mutation of any gene were detected. Based on these data, we recommend that ELA2 genotyping be performed in all patients with suspected SCN. In the North American population mutations in HAX1, GFI1, SBDS, and WAS are rare and routine genotyping is not indicated. Finally, the data suggest that there are yet undiscovered genetic causes of SCN.
Gfi-1 Represses CDKN2B Encoding p15INK4B through Interaction with Miz-1
Authors: Liu Qingquan , Basu Suchitra , Qiu Yaling , Dong Fan 
Published: October 2019

Abstract
Zinc finger (ZF) transcriptional repressor Gfi-1 plays an important role in hematopoiesis and inner ear development, and also functions as an oncoprotein that cooperates with c-Myc in lymphomagenesis. Gfi-1 represses transcription by directly binding to the consensus DNA sequence in the promoters of its target genes. We report here an alternative mechanism by which Gfi-1 represses CDKN2B encoding the cyclin-dependent kinase inhibitor p15INK4B. Gfi-1 did not directly bind to CDKN2B, but interacted with Miz-1 and, via Miz-1, was recruited to the core promoter of CDKN2B. The C-terminal zinc finger domains of Gfi-1 and Miz-1 are involved in the interaction. Miz-1 is a POZ-ZF transcription factor that has been shown to mediate transcriptional repression by c-Myc. Like c-Myc, upon recruitment to the CDKN2B promoter, Gfi-1 repressed transcriptional activation of CDKN2B by Miz-1 and in response to TGFb. Notably, Gfi-1 and c-Myc formed a ternary complex with Miz-1 and were both recruited to the CDKN2B core promoter via Miz-1, and acted in collaboration to repress CDKN2B. Consistent with its role in repressing CDKN2B transcription, knockdown of Gfi-1 in human leukemic cells resulted in augmented levels of p15INK4B, which was associated with attenuated cell proliferation. The expression of p15INK4B was also significantly higher in Gfi-1−/− mouse bone marrow (BM) cells than in Gfi-1+/+ BM cells. Our data reveal a novel mechanism of transcriptional repression by Gfi-1 and also identify CDKN2B as a new Gfi-1 target gene. The findings may have important implications for understanding the role of Gfi-1 in normal development and the cooperation between Gfi-1 and c-Myc in lymphomagenesis.
Random Mutagenesis Reveals Residues of JAK2 Critical in Evading Inhibition by Tyrosine Kinase Inhibitors
Authors: Marit Michael , Chohan Manprit , Matthew Natasha , Huang Kai , Barber Dwayne 
Published: October 2019

Abstract
Mutation and activation of JAK2 is a common event in Myeloproliferative Disease as JAK2 V617F and related deletion mutations are observed in Polycythemia Vera, Essential Thrombocythemia and Primary Myelofibrosis. In addition, the TEL-JAK2 chromosomal translocation is a rare event in Acute Myeloid Leukemia and Chronic Myelomonocytic Leukemia. We reasoned that activated alleles of JAK2 would develop resistance to JAK inhibitors in a clinical setting, similar to the development of Imatinib resistance in BCR-ABL-mediated Chronic Myeloid Leukemia. The objective of this study was to develop a random mutagenesis screen to isolate mutations of JAK2 resistant to tyrosine kinase inhibitors. We selected JAK Inhibitor-1 for this study, since the crystal structure of this inhibitor bound to the JAK2 JH1 kinase domain has been reported and would allow for mapping of confirmed mutations. TEL-JAK2(5–12) and JAK2 V617F were subcloned into retroviral expression vectors and random libraries of mutations were generated by transformation into XL-1 Red strain of E. coli, a strain defective in pathways of DNA repair. High titer retroviral supernatants were generated and used to transduce Ba/F3 (for TEL-JAK2) or Ba/F3-EPO-R (for JAK2 V617F). Control experiments were performed with “wild type” versions of each JAK2 allele. Inhibitor-resistant clones were identified and DNA sequencing was performed to identify JAK2 JH1 kinase domain mutations that confer resistance to inhibitor. We have restricted the analysis of JAK inhibitor-resistant mutants to those that map within the kinase domain of JAK2 for purposes of this study. We have confirmed that E864K, V881A, N909K, G935R, R975G confer resistance to JAK inhibitor-1 in growth assays. In addition, M929I (analogous to BCR-ABL T315I) mediates resistance to JAK inhibitor-1, relative to wild-type activated JAK2 alleles. All mutations result in increased phosphorylation of STAT5, Akt and Erk in the presence of inhibitor. We are currently testing the catalytic activity of each mutant to determine whether JAK2 kinase domain mutations have similar enzymatic activity or whether mutation also affects catalysis. We have mapped each mutation within the JAK2 JH1 crystal structure and models for how each mutant affects inhibitor binding will be presented. Importantly, many of these residues are highly conserved in other JAK tyrosine kinases and within BCR-ABL. We are extending these observations by testing clinically relevant inhibitors in our screen. At the conclusion of this study we will be able to identify common residues critical for resistance by JAK2 inhibitors and unique residues that are inhibitor-specific. Random mutagenesis screening offers an excellent strategy to identify JAK2 residues that may be relevant in the clinic and also serve in enhancing our knowledge regarding JAK kinase activation and regulation.
Immune-Related and Inflammatory Conditions Likely Play a Role in the Development of Lymphoplasmacytic Lymphoma/Waldenström’s Macroglobulinemia
Authors: Kristinsson Sigurdur Y , Bjorkholm Magnus , Koshiol Jill , Goldin Lynn R. , McMaster Mary L. , Turesson Ingemar , Landgren Ola 
Published: October 2019

Abstract
Introduction: Certain autoimmune and infectious conditions are associated with increased risks of subtypes of non-Hodgkin lymphomas (NHL). A few prior studies suggest that chronic immune stimulation may particularly elevate risk for the NHL subtype lymphoplasmacytic lymphoma (LPL)/Waldenström’s macroglobulinemia (WM). To improve our understanding on the role of immune-related and inflammatory conditions in LPL/WM pathogenesis, we conducted a large population-based study including close to 2,500 LPL/WM patients diagnosed in Sweden and almost 10,000 matched controls.
Methods: Using both the central Cancer registry and local hospital-based registries, we identified all LPL/WM patients diagnosed in Swedish hospitals 1958–2005. From the Swedish Population Registry we identified four matched controls per LPL/WM patient. Through data linkage with the central Inpatient registry, we gathered information on hospital inpatient discharges that listed autoimmune-, infectious-, and other inflammatory/allergic diseases present at least 1 year prior to LPL/WM. Using Poisson regression, we calculated rate ratios (RR) and 95% confidence intervals (CI) adjusted for categorical year of birth, date of diagnosis, gender, and county.
Results: A total of 2,470 LPL/WM patients (647 LPL and 1,823 WM), and 9,698 population-based matched controls were included in the study. We found an increased risk of developing LPL/WM among individuals with a prior history of systemic sclerosis (RR=4.7; 1.4–15.3), Sjögren’s syndrome (RR=12.1; 3.3–45.0), autoimmune hemolytic anemia (AIHA) (RR=24.2; 5.4–108.2), polymyalgia rheumatica (RR=2.9; 1.6–5.2), and temporalis arteritis (RR=8.3; 2.1–33.1). We also found excess risk of LPL/WM among persons with a history of pneumonia (RR=1.4; 1.1–1.7), septicaemia (RR=2.4; 1.2–4.3), pyelonephritis (RR=1.7; 1.1–2.5), sinusitis (RR=2.7; 1.4–4.9), herpes zoster (RR=3.4; 2.0–5.6), and influenza (RR=2.9; 1.7–5.0). Importantly, when we assessed the associations by latency (time between immune-related or inflammatory conditions and LPL/WM), for most autoimmune- and infectious diseases the excess LPL/WM risk remained significant at >5 years latency. We found no significant increased risk for LPL/WM among individuals with prior chronic inflammatory or allergic conditions.
Conclusions: In the largest investigation of risk factors for LPL/WM to date, we found a personal history of certain autoimmune and infectious diseases to be associated with excess LPL/WM risk. Immune-related conditions might act as potential LPL/WM triggers or they could represent premalignant immune disruptions preceding LPL/WM. Our results provide novel insights into the as yet unclear pathogenesis of LPL/WM.