bluuebunny/crossref_metadata_2025_split huggingface dataset:
Promoter Methylation of Multiple Genes in Germinal Center Hyperplasia and Lymphomas of Germinal Center Origin
Authors: Paz-Carreira Jose M , Losada Raquel , Garcia-Rivero Arantxa , Alvarez Augusto , Bal Fernando , Alba Jesus 
Published: April 2020

Abstract
INTRODUCTION. Germinal centers (GC) are unique sites in peripheral lymphoid tissue where clonal selection of B cells takes place. This occurs as a response to stimulation by various antigens originating, sometimes, follicular hyperplasia (FH). GC have been known as a major source of B-cell lymphomas including follicular (FL) and diffuse large cell (DLCL). DNA methylation of tumor-suppressor genes is a mechanism of gene silencing involved in the pathogenesis of FL and DLCL. Much less is known about the role of methylation in FH. We determined the methylation status of 6 tumor-suppressor genes in 43 patients with FH, 18 patients with FL and 49 patients with DLCL in order to see the differential implication of this epigenetic mechanism in the pathological and the physiological development of GC.
MATERIAL AND METHODS. Genomic DNA extracted from paraffin-embedded samples of 43 FH, 18 FL and 49 DLCL after being treated with EZ DNA-Methylation Kit (Zymo Research) with the manufacturer’s instructions, were analyzed by methylation-specific polymerase chain-reaction to determine promoter hypermethylation of DAP-k, SHP1, Rarβ, p14, SHP1, MGMT and PDRM1. All samples were obtained mostly from lymph nodes and tonsils. Diagnosis was based on morphology and immunohistochemistry analysis. All cases were matched for age, sex and ethnic origin.
RESULTS: DAP-k promoter methylation occurred with higher frequency in FL(89%) than in DLCL(78%) and FH(40%). SHP1 was methylated in 61% of FL, 58% of FH and 23% of DLCL. RARb was methylated in 67% of FL patients, 30% of DLCL and only 12% of FH. Eight (44%) FL, seventeen (35%) DLCL and four (10%) BFH patients showed MGMT methylation. Promoter hypermethylation of p14 was detected only in 5 (12%) FH, 2 (4%) DLCL and none FL patients. Methylation of PRMD1 was present only in 1 (6%) FL, 2 (6%) DLCL and 1 (4%) FH samples.
CONCLUSIONS. Inactivacion of DAP-K and SHP1 is present in B-cell malignancies, DLCL and FL, and BFH. Therefore, it may represent a physiologic event conferring a temporal survival advantage necessary for a GC hyperplastic response. Inactivation of the retinoic acid response through the methylation of Rarâ is significantly more frequent in lymphomas than in FH. As reported in other tumors methylation of MGMT is more frequent in lymphomas than in FH. With our data methylation of Cyclin dependent kinase inhibitors p14 is not a differential pathogenic event in lymphomas of GC origin, in fact it is more frequent in FH. Promoter Methylation of PDRM1 is not the mechanism involved in lymphomagenesis in FL and DLCL, the two FH positive deserve further follow-up to determine its significance.
Expression of NFATc1 in Diffuse Large B Cell Lymphomas (DLBCL) Is Decreased in HIV Positive Patients
Authors: Homan S. M , Patel R. A , Sheehan C. E , Hayner-Buchan A. , Ross J. S , Nazeer T. 
Published: April 2020

Abstract
Background: DLBCL is the most common non-Hodgkin lymphoma (NHL), comprising 30 to 40% of adult NHLs. It may present with nodal or extranodal involvement or it may be primary to the central nervous system (CNS). HIV positive patients comprise a significant percentage of patients diagnosed with DLBCL, however, differences in molecular characteristics based on HIV status have not been elucidated. Understanding the molecular basis for lymphomas is integral to the development of targeted therapy. The NFAT (nuclear factor of activated T cells) family is a group of cytoplasmic transcription factors that undergoes nuclear translocation when activated and plays an important role in intracellular signaling. This study evaluated the differential expression of the NFATc1 protein in HIV positive and negative patients with DLBCL (nodal, extranodal and primary CNS).
Design: Formalin-fixed, paraffin-embedded archival tissue from 34 nodal/extranodal DLBCLs and 41 primary CNS DLBCLs were immunostained with antibodies against NFATc1 by automated methods (NFATc1 (7A6):sc-7294, Santa Cruz Biotechnology, Inc, Santa Cruz, CA;Ventana Medical Systems, Inc, Tucson, AZ). Nuclear immunoreactivity in neoplastic tissue was semiquantitatively assessed and the results were correlated with HIV status and overall patient survival.
Results: Nuclear NFATc1 immunoreactivity was noted in both nodal/extranodal and primary CNS DLBCLs with 13/34 (38%) nodal/extranodal DLBCLs and 13/41 (32%) primary CNS DLBCLs showing nuclear NFAT1c positivity. Of the 59 total HIV negative cases, 42% (25/59) showed nuclear NFAT1c immunoreactivity. In contrast, less than 1% (1/16) of HIV positive cases demonstrated nuclear NFAT1c immunoreactivity (p=0.007). Within the nodal/extranodal DLBCL subgroup, nuclear NFATc1 positivity was observed in 12/23 (52%) HIV negative cases versus 1/11 (9%) HIV positive cases; while within the primary CNS subgroup nuclear positivity was noted in 13/36(36%) HIV negative cases versus 0/5(0%) HIV positive cases. There was no correlation with overall survival in either subgroup.
Conclusions: Similar to previous studies, nuclear NFAT1c immunoreactivity was demonstrated in DLBCLs and primary CNS lymphomas. Interestingly, this nuclear NFAT1c immunoreactivity was significantly decreased among patients with HIV positive status suggesting activation of alternative intracellular signaling pathways. These findings highlight potential variations in gene expression among DLBCLs based on HIV status. This decreased expression of NFAT1c amongst HIV positive individuals may be of particular importance when developing targeted therapy for aggressive B-cell malignancies.
Phasee I Biological Study of IV and SQ Azacitidine (Vidaza®) to Determine the Optimal Biological Dose in Hematologic Malignancies
Authors: Yang Allen S. , Mohrbacher Ann , Douer Dan , Gorospe Gerry , Byun Hyang-Min 
Published: April 2020

Abstract
Azacitidine (5-azacytidine, Vidaza) is a DNA methylation inhibitor that has been shown to have a survival benefit in patients with high-risk myelodysplastic syndrome and is under investigation in other cancers. The purpose of this study was to determine the optimal dose and route of administration for azacitidine to inhibit DNA methylation in patients with hematologic malignancies. Eligibility included patients with hematologic malignancy who provided informed consent. Enrollment criteria varied depending on the type of cancer. Patients were enrolled into 5 escalating dose levels for the first course of therapy (25mg, 50mg, 75mg, 100mg or 150mg IV per m2 per day for 5 days). On day 28 all patients received 75mg/m2/day IV for 5 days. For course 3 patients received 75mg/m2/day SQ × 5 days. Beyond course 3 patients received 75mg/m2/day either SQ or IV × 5 days every 4 weeks. Peripheral blood was collected on days 1, 3 and 5 for each course and LINE1 DNA methylation was measured using bisulfite-PCR Pyrosequencing to measure global DNA methylation changes induced by azacitidine. To date, 17 patients have been treated (3 at 25mg, 4 at 50mg, 4 at 75mg, 3 at 100mg, and 3 at 150mg/m2). Diagnosis included 6 patients with MDS, 9 patients with AML (3 untreated older patients, 6 relapsed or refractory patients), 1 patient with CML (imatinib refactory), and 1 patient with non-Hodgkin’s lymphoma (relapsed disease). At the time of submission (August 2008) 12 patients were evaluable for response with 1 CR, 3 PR, 7 SD and 1 PD reported. The median number of cycles given was 2 (range 0–12+). LINE1 DNA methylation decreased by 1.4, 2.3, 4.8, 1.9 and 4.0% on day 5 for the 25mg, 50mg, 75mg, 100mg, and 150mg/m2 dose levels respectively. Mean decrease in LINE1 DNA methylation with 75mg/m2 IV was 3.7% and only 2.6% by 75mg/m2 of azacitidine SQ. There was a large amount of inter-patient variability but less intra-patient variability in DNA methylation response to azacitidine. DNA methylation and data analysis is ongoing. Azacitidine is effective at inhibiting DNA methylation at multiple dose levels for both IV and SQ routes of administration. There is a high degree of patient-to-patient variability in DNA methylation changes, but 75mg/m2 lead to the greatest mean decrease in DNA methylation by a 5 day IV regimen. Updated analysis for this study will be presented.
Acquired Multiple Coagulation Factor Inhibitors Associated Bleeding Disorder
Authors: Chen Qin-fen , Li Pei , Xie Yi , Chen Bao-An 
Published: April 2020

Abstract
Objective: To inquire into the clinical features of acquired multiple coagulation factors inhibitors associated bleeding disorder.
Methods: A case of acquired multiple coagulation factors inhibitors in clinical manifestations, diagnosis, treatment and result was described and related literatures were reviewed.
Results: A 74-year-old man developed sustained wound bleeding after implantation of heart pacemaker. Prothrombin time (PT), activated partial thromboplastin time (APTT) were prolonged significantly. But thrombin time (TT), fibrinogen (Fg), platelet count(PLT) and liver function were normal. 3P test was negative. Further tests revealed that the activities of factor 2, 5, 7,8,9,10,11,12 were all less than 10%, and the inhibitors of these factors could be detected, with the titers ranging from 8~64 Bu. So acquired multiple coagulation factor inhibitors was diagnosed. Transfusion with frozen plasma and cryoprecipitate was ineffective, whereas the combination therapy with glucocorticoid plus plasma exchange seemed to be successful. The patient was cured.
Conclusion: Acquired multiple coagulation factors inhibitors is a rare bleeding disorder. In fact, this is the first case, as far as we are aware. It may develop serious bleeding. Immunosuppressive agents, such as corticosteroids, used for suppression of autoantibodies formation and plasma exchange, used for eradication of inhibitors may have beneficial effects.
Radiosynovectomy in Children with Congenital Haemostatic Defects
Authors: Laguna Pawel PL , Zbikowski Piotr , Cwikla Jaroslaw , Klukowska Anna , Walecki Jan , Matysiak Michal 
Published: April 2020

Abstract
Aims: To evaluate the clinical benefits of radiosynovectomy (RS) in children with congenital coagulative disorders, reduce bleeds and improve progression free survival (PFS).
Methods: 35 patients aged 7 to 17 yrs, (mean age 12.6 yrs) were included in the study. 29 had haemophilia A, (4 with inhibitor factor VIII), 3 had haemophilia B (1 with inhibitor factor IX) and 3 had von Willebrand’s disease. There were 43 therapies including injections into the followed joints: knee 21, elbow 13, ankle 6 and shoulder 3. In 30 cases this was initial therapy and in 5 cases repeat therapy due to recurrence or deterioration. Injected activity of 186Re colloid was dependent on type and size of joints, range: 60–180 MBq. Post therapy imaging was performed in each case 1 h after injection of radioisotopes, and then on day 2 and 3. The number of bleedings into joints per month, improvement of joint movements, and other clinical changes were evaluated before and after therapy in 3 month intervals. US images were used to assess changes in joint effusion and synovial overgrowth. PFS in terms of significant clinical improvement within joints was assessed using standard Kaplan Meier methods. The prognostic significance of selected parameters was tested using discriminate function analysis.
Results: After injection of 186Re into joints, post-therapeutic images showed intraarticular distribution of tracer and there was no leakage. A significant reduction in bleeds into joints was noted (before therapy mean 1.8 per month and after treatment mean 0.3 per month, p&lt;0.001).
Conclusions: This study suggests that use of 186Re in early stages of arthropatic haemophilia reduces the number of bleeds into joints and improves PFS.
Increased Plasminogen Activity Is An Independent Predictor of Bleeding in Hemophiliacs
Authors: Gerhardt Andrea , Pesch Melanie , Scharf Rudiger E. , Zotz Rainer B. 
Published: April 2020

Abstract
Rationale and Objectives: Apart from the differences in factor VIII:C levels, it is difficult to explain the significant variation in bleeding events observed among individual patients with similar levels of FVIII:C and standard laboratory profiles. We hypothesize that additional dispositional (genetic) and expositional (non-genetic) factors of hemostasis may augment or attenuate the likelihood of bleeding symptoms in hemophiliacs.
Patients and Methods: To test this hypothesis, we studied 61 patients diagnosed with hemophilia A. The following laboratory parameters were examined in multivariate analysis using a case-only analysis: genotyping for a2C807T, HPA-1 of aIIbb3, factor V G1691A, prothrombin G20210A; coagulation factor activity/antigen of fibrinogen, factor II, V, VII, VIII, IX, X, XI, XII, XIII, vWF; in-vitro bleeding time (PFA-100 closure times), and plasminogen.
Results: After adjustment for age, low levels of factor VIII (p=0.0296) and increased levels of plasminogen (p=0.0415), and collagen/ADP closure times (PFA-100) (p=0.04) could be identified as independent predictors of mucosal bleeding. 10% higher activity of plasminogen increased the bleeding risk 10.4 fold. For joint bleeding low levels of fibrinogen (p=0.003) and longer collagen/epinephrine closure times (PFA-100) could be identified as bleeding predictors.
Conclusion: Apart from low levels of factor VIII, increased activity of plasminogen is an independent predictor of mucosal bleeding in hemophiliacs. In consequence, it will be of importance to examine whether the critical subgroup of patients with increased plasminogen levels and mucosal bleeding can benefit from prevention with specific antifibrinolytic agents.
Interferon Gamma +874A/T Polymorphism in Children with Idiopathic Thrombocytopenic Purpura
Authors: Demircioglu Fatih , Ören Hale , Kizildag Sefa , Yilmaz Sebnem , Atabay Berna , Turker Meral , Vergin Canan , Irken Gulersu 
Published: April 2020

Abstract
A recent study showed that expression of Toll-like receptor and interferon-gamma associated genes is significantly increased in patients with chronic ITP. Interferon-gamma is an important protein which takes place in immunoregulation. +874A/T polymorphism in the first introne of interferon gamma gene is found to be associated with the development and clinical phenotype of some autoimmune diseases such as diabetes mellitus, thyroiditis, multiple sclerosis, and SLE. The aim of our study was to investigate whether interferon gamma +874A/T polymorphism is a risk factor for the development of ITP and whether it affects the clinical course and response to the treatment. Thirty five children with acute ITP and 40 children with chronic ITP who were followed for at least 6 months were included. Control group consisted 90 healthy children. Two millilitres of blood sample was taken into sterile tubes containing 0.1% EDTA from each child and all blood samples were stored at −20 until analysis. DNA was isolated from blood samples and interferon gamma +874A/T polymorphism was studied with real-time PCR and LightCycler TM. Twenty one patients had AA, 35 patients had AT, and 19 patients had TT genotype. In the control group, 47 children had AA, 36 children had AT, and 7 children had TT genotype. There was a statistical difference between ITP and control group regarding the genotype (p=0.001). The frequency of A and T alleles in ITP group was 52% and 48%, respectively. The frequency of A and T alleles in control group was 72.7% and 27.8%, respectively. The frequency of allele distribution was statistically different between the ITP and control groups (p&lt;0.0001). There was a statistical significant difference between acute ITP and control group regarding the frequency of AA, AT, and TT gene polymorphisms and allele frequency (p=0.002, p=0.002). Similarly, there was a statistical significant difference between chronic ITP and control group regarding the frequency of AA, AT, and TT gene polymorphisms and allele frequency (p=0.008, p=0.002). The frequency of AA, AT, and TT gene polymorphisms and allele frequency showed no statistical difference between acute and chronic ITP groups (p=0.285, p=0.896). There was no correlation between interferon gamma +874A/T polymorphism and severity of bleeding (mild, moderate and severe) (p=0.09). There was no correlation between interferon gamma +874A/T polymorphism and response to long term treatment in patients with chronic ITP (p=0.568). In conclusion, there was a significant difference between patients with ITP and children in control group regarding interferon gamma +874A/T polymorphism and in the light of recent data involving other autoimmune disorders, we think that interferon gamma +874A/T polymorphism may be a risk factor for ITP.
Identification of Three Novel Substitutions in the Erythroid Promoter of the HMBS Gene: A New Erythroid Variant of Human Acute Intermittent Porphyria?
Authors: Di Pierro Elena , Besana Valeria , Brancaleoni Valentina , Tavazzi Dario , Cappellini Maria Domenica 
Published: April 2020

Abstract
Porphyrias are inherited disorders in the biosynthesis of heme. Acute intermittent porphyria (AIP), the most common form, is autosomal dominant and it is characterized by recurrent attacks of abdominal pain, gastrointestinal dysfunction, and neurologic disturbances. AIP is caused by molecular defects in the hydroxymethylbilane synthase gene (HMBS) causing a partial deficiency of the third enzyme of the heme biosynthetic pathway. This gene maps to chromosome 11q23.2 with a total of 15 exons. Two distinct promoters direct the synthesis of housekeeping and erythroid specific mRNAs by alternative splicing. The housekeeping promoter is located upstream the exon 1 while the erythroid promoter include a portion of intron 1, the exon 2 and the intron 2. The exon 1 and the exons 3 to 15 generate the housekeeping mRNA while the exons 2 to 15 generate the erythroid mRNA giving rise to the housekeeping HMBS isoform and to the erythroid HMBS isoform respectively. In the classic form of AIP, both the housekeeping and the erythroid HMBS isoforms are deficient since the must common molecular defects involve the common region of the gene from exon 3 to exon 15. Approximately 5% of AIP patients have normal levels of the erythroid HMBS isoform and defects in the housekeeping promoter or in the exon 1 which is specific for the housekeeping HMBS isoform, causing the ‘non-erythroid variant’ of AIP. So far no mutations are known in the erythroid promoter. In this study we searched for molecular defects in HMBS gene in three Italian patients with typical clinical and biochemical signs of AIP. The diagnosis was based on elevated urinary excretion of porphyrinic precursors and reduced erythrocyte HMBS activity. The entire HMBS gene has been amplified by PCR and submitted to direct automated sequencing: no mutations known as responsible for classic form of AIP have been identified. However, each of three patients had a substitutions in the erythroid promoter of HMBS gene that could justify the reduced erythrocyte HMBS activity: c.34–115 C&gt;A, c.34–126 G&gt;C and c.34–156G&gt;A. In order to establish if these substitutions were polymorphisms, more than 200 alleles from Italian normal subjects were sequenced and none of them revealed the substitutions. These three mutations, located specifically in the intron 2, could affect the normal splicing of exon 3 causing an abnormal mRNA, to give rise to the classical form of AIP. The mRNA analysis, however, didn’t reveal any abnormal mRNA. Moreover real time experiments in the lymphocytes showed a 100% expression of housekeeping HMBS mRNA excluding the non-sense mediated decay mechanism. These data suggest that these mutations could affect the normal function of the erythroid promoter only, causing an erythroid variant of human acute intermittent porphyria. Further expression studies are in progress.
Effects of down-Regulating BCL11B Expression on the Proliferation and Apoptosis of Molt-4 Cells by RNA Interference
Authors: Chen Si , Li Yangqiu , Huang Xin , Yang Lijian , Chen Shaohua , Zhou Yubing 
Published: April 2020

Abstract
The B-cell chronic lymphocytic leukemia (CLL)/lymphoma 11B (BCL11B /CTIP2/ RIT1/hRitα) gene, which encodes C2H2 zinc-finger transcription factors, plays critical roles during development of T lymphocytes and the central nervous system. Recently, BCL11B gene has been identified to be associated with hematopoietic malignancies, such as T-cell acute lymphoblastic leukemia (T-ALL). However, little is known regarding the molecular mechanism by which BCL11B may contribute as a tumor-suppressor gene. In order to clarify the role of BCL11B gene in T-cell malignancies, we analyzed the effects of BCL11B target-specific small interfering RNA (siRNA) on the proliferation and apoptosis of Molt-4 cells, which is the naturally BCL11B overexpressing cell line. BCL11B-specific siRNA and scrambled non-silencing siRNA control were designed and synthesized by Invitrogen. The siRNAs were transfected into MOLT-4 cells by nucleofector techique. The level of BCL11B mRNA expression was analyzed at 48 and 72 hours after nucleofection by Real-time quantitative PCR with Taqman technique. The cells proliferation rate was assayed by MTT method at 24 and 48 hours after transfection, respectively. Cell morphology was showed by Liu’s Stain. The changes of cell cycle and apoptosis rate were detected by flow cytometry. The results showed that the level of BCL11B mRNA expression was inhibited in siRNA transfected cells at 72 hours after transfection, as compared with scrambled controls. The survival rates of siRNA-transfected cells were significantly decreased at both 24 and 48 hours (P &lt; 0.01). Apoptotic cells could be observed morphologically. The apoptosis rates of Molt-4 cells treated with BCL11BsiRNA were highly increased compared with scrambled control ( P &lt; 0.05). However, the percentage of different phases did not show significantly difference between siRNAtransfected cells and control cells, which was suggested that the apoptosis could be induced all phases of cell cycle. In conclusion, our data indicated that, the survival of Molt-4 cells might be critically dependent on BCL11B gene. Downregulation of BCL11B expression by RNA interference selectively can inhibit the proliferation and induced the apoptosis effectively in transformed Molt-4 cells, farther investigation will be performed in primary T-ALL cells to confirm the effect of presented BCL11B-siRNA on T-ALL.