bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression of Circadian Core Clock Gene Per2 Is Markedly Down-Regulated in Diffuse Large B-Cell Lymphoma
Authors: Thoennissen Nils Heinrich , Cao Qi , Doan BaoNgan , Abbassi Sam , Nowak Daniel , Müller-Tidow C. , Said Jonathan , Gary Sigal , Koeffler H. Phillip 
Published: April 2020

Abstract
Non-Hodgkin’s lymphoma (NHL) is a heterogeneous clinicopathologic entity characterized by distinct cells of origin, cytogenetic and molecular aberrations. In vivo studies recently showed that mice deficient in Period (PER) 2 developed lymphomas. Furthermore, we and others have recently suggested that the core circadian clock genes Period (Per)1 and Per2 have been linked to DNA damage response pathways. We showed that both genes are involved in tumor suppression by regulating cell cycle- and apoptosis-related genes. In mammals and other vertebrates, heterodimers of the basic helix-loop-helix-PAS transcription factors, CLOCK and BMAL1, activate transcription of the Period and Cryptochrome (Cry1 and Cry2) genes via binding to E-box sequences in their promoters. Subsequently, the PER and CRY repressor proteins accumulate in the nucleus and inhibit CLOCK:BMAL1 complexes, thereby inhibiting their own gene expression. This forms the major negative circadian feedback loop. In the present study, we focused on Per1 and Per2 and their role in different types of NHL. Real-time reverse transcriptase-polymerase chain reaction (real time RT-PCR) was performed to determine the mRNA expression levels of Per1 and Per2 in patients with diffuse large B-cell lymphoma (DLBCL; n = 37), mantle cell lymphoma (MCL; n = 12), follicular lymphoma (FL; n = 12), and Burkitt’s Lymphoma (n = 6) compared to human normal tonsil samples (n = 10). We further tested their expression in 14 different human lymphoma cell lines. The relative expression of Per2 was markedly down-regulated in all DLBCL samples compared to normal tonsils (mean fold change: − 20.5 ± 28.7; p &lt; 0.001). On the other hand, levels of expression of Per2 in samples of either MCL (2.5 ± 2.6; p = 0.1), FL (1.1 ± 10; p = 0.9), or Burkitt’s Lymphoma (− 1.1 ± 2; p = 0.07) showed no significant change compared to the normal tonsils. Moreover, we found no significant regulation of the mRNA levels of Per1 in any of the four NHL subtypes. We confirmed our results by showing concordant results in human lymphoma cell lines. Again, Per2 expression levels were persistently down-regulated in the 7 DLBCL cell lines (OCI-Ly1, -Ly4, -Ly7, -LY10, and SUDHL-4, -6, -16; mean fold change: − 8.6 ± 5.7; p &lt; 0.001) compared to normal tonsils, but not in MCL (SP-49, Jeko-1, NCEB-1), FL (FLK-1), Burkitt’s lymphoma (Daudi), and B-precursor acute lymphoblastic leukemia (Nalm-6, BALL-1) cell lines (p &gt; 0.05). Western blot protein analyses showed easily detectable levels of PER2 in tonsil samples, but the protein was either not expressed or significantly reduced in expression in human DLBCL cell lines. In summary, our results strongly suggest for the first time that the disruption of the key circadian clock gene Per2 may play a role in the development of DLBCL.
The Research of Phenylhexyl Isothiocyanate’s Influence on p16 Gene Methylation of Leukaemia Cell
Authors: Chen Bao-An , Shou Bei-ming , Zhou Dong-rui , Liu Delong , Ding Jia-hua , Sun Yun-Yu , Wang Jun , Cheng Jian , Zhao Gang , Song Hui-hui , Bao Wen , Wang Jue-qiong , Yuan Peng , Pei Xiao-ping , Chen Ning-na , Ma Xu-dong , Lu Zu-hong , Lu Quan-yi , Lai Yong-rong , Huang He 
Published: April 2020

Abstract
Objective Aberrant DNA methylation of CpG site is among the earliest and most frequent alterations in cancer. The aim of this study was to study whether phenylhexyl isothiocyanate can reduce the methylation level of P16 gene. This study used a microarray-based method for quantificationally detecting changes of P16 gene methylation in leukemia patient and U266 cell line. And to simply discuss the effect of phenylhexyl isothiocyanate on tumor methylation.
Methods This method used bisulfite-modified DNA as a template for PCR amplification, resulting in conversion of unmethylated cytosine, but not methylated cytosine, into thymine within CpG islands of interest. One set of oligonucleotide probes were designed to fabricate a DNA microarray to detect the methylation changes of P16 gene CpG islands. Each set contained a pair of methylated and unmethylated oligonucleotides for interrogating 3 CpG sites in close proximity. By TA cloning, PCR, sequencing, positive and negative DNA targets were required. Next drawing a standard curve by fluorescence intensity. Leukemia samples DNA were abstracted and bisulfite-modified. Sample DNA targets were required by PCR amplification and were hybridized with the microarry. Finally the microarry was scanned with ScanArray Lite microarray analysis systems.
Results The linear relationship (R2=0.9660) was established and it could be used to eliminate background noise. The methylation level of U266 is hypermethylation, after cultured with PHI, the level reduce. Eleven patients have P16 gene hypermethylation in thrity. After culture with PHI, seven patients showed level reduce, one patient showed raise, three patients showed remaining.
Conclusion PHI can reduce the methylation level of P16 gene in U266 cell line, and also can reduce the methylation level of P16 gene of leukemia patient samples in vitro culture.
Rare Congenital Bleeding Disorders from a Tertiary Haematology Referral Center in Western India
Authors: Kelkar Anjali Jaydeep , Apte Shashikant , Melinkeri Varsha , Melinkeri Samir , Karpe Ashay , Bhave Saurabh , Sayaji Bhavna 
Published: April 2020

Abstract
Introduction: Bleeding disorders are present in varying frequency in a given population. We present our data of rare bleeding disorders (excluding Haemophilia & von Willebrand’d disease) since little data about the same is available from developing countries. Many of the rare bleeding disorders are associated with a strong history of consanguinity, the frequency of consanguineous marriages are between 12 –42 %.
Materials and Methods: These cases were part of the referral for evaluation of congenital bleeding disorders; some of the cases were diagnosed as part of pre-operative coagulation screen. All the cases were subject to a detailed history encompassing the timing site and frequency of bleed, history of consanguinity, umbilical cord bleed. Lab. evaluation included bleeding time (Ivy’s method), clotting time., in vivo adhesion and clot retraction. Prothrombin and activated partial thromboplastin time with correction studies as and when required, Factor XIII screen with urea solubility test. Complete blood count was done prior to bleeding time and platelet adequacy was confirmed. All blood samples used for coagulation assays were collected in sterile citrate vaccutainers. Further investigations were performed including factor assays depending upon the results of the basic profile.
Results: Feb 2000 till August 2008.
Total No. of cases studies I from Year 2000 970 Abnormal cases 75 Consanguinity 25 Disorder No. Of Cases Factor V Deficiency 5 Factor VII Deficiency 4 Factor X Deficiency 4 Factor XI Deficiency 1 Factor XII Deficiency 6 Factor XIII Deficiency 8 Combined factor deficiency 4 Factor I deficiency 11 Dysfibrinogenemia 6 Factor II deficiency 1 Glanzmann’s thrombasthenemia 25 Bernard soullier’s syndrome 2
Conclusion: In our institution which caters to a large population in and around city of Pune, Maharashtra Western India, we have detected a good number of patients with rare bleeding disorders. These disorders can be found on a more frequent basis as we have a significant incidence of consanguineous marriages, as our data shows i.e approx. 33%. This report highlights need for extended family search and molecular characterization so as to make antenatal diagnosis amenable for severe bleeding disorders.
Use of Factor VIII/Von Willebrand Factor Complex for Inducing Immune Tolerance in Hemophilia a Patients Who Have Failed with Recombinant Fviii: Recent Canadian Experience
Authors: Vincent Anne-Marie , Chan Anthony , Israels Sara J. , Poon Man-Chiu , Wu John K. , Rivard Georges E. 
Published: April 2020

Abstract
Immune tolerance induction (ITI) is effective in approximately 63–83% of hemophilia patients with inhibitors. Poor prognostic factors are age &gt; 18 years, ITI started &gt; 4 years after inhibitor development, inhibitor peaks &gt; 200 BU, inhibitor titre &gt; 10 BU at ITI initiation, and previously failed ITI. It has been reported that patients who have failed ITI with a recombinant FVIII concentrate achieved and maintained immune tolerance when VWF/FVIII complex concentrates were used. We present our Canadian experience with a factor VIII/von Willebrand factor concentrate (Humate-P) for inducing immune tolerance in patients who have previously failed ITI. A questionnaire was sent to all Hemophilia Treatment Centers of Canada. Data collected included baseline FVIII level, mutation, date at initial diagnosis of inhibitor, peak titer, concentrate associated with development of inhibitor, inhibitor titer at initiation of first ITI, concentrate and regimen used, and concentrate, dosing and outcome of secondary ITI with Humate-P. All Hemophilia Centers agreed to participate. 5 Centers submitted data about 8 patients. One patient resistant to initial ITI with Kogenate FS achieved good partial response with Humate-P, with titers going down from 40 to values between 0,6 and 4 BU. One patient who responded to initial ITI with Kogenate FS with peaking of inhibitor to 380 BU was remaining above 100 BU and lowered to 28 BU on Humate-P. In three patients, Humate-P induced an anamnestic response. Two of those patients had a concomitant line infection at time of switching to Humate-P. The third of those three patients had a transient anamnestic response after introduction of Humate-P. One patient had successfull induction of immune tolerance using multiple therapies (cyclophosphamide, Immunate, IVIG, Rituximab) and maintained remission under Humate-P. One patient was on ITI for 27 months using Recombinate and switched to Humate-P when Recombinate was withdrawn from the market. After 12 months, the inhibitor had gone from 12 to 7.3 BU. One patient attained complete remission during primary ITI using recombinant FVIII concentrate (Advate) and maintained remission when switched to Humate-P. In subjects failing ITI with recombinant factor VIII concentrates, Humate-P may be considered as a second line for inducing immune tolerance.
Incidence of Suicide in U.S. Hemophilia Patients
Authors: Gomperts Edward D. , Presley Rodney J , Baker Judith , Holtz Jean 
Published: April 2020

Abstract
The causes of death in hemophilia have changed in the USA over the past few decades. Prior to the 1970’s, individuals with hemophilia were succumbing to the seriousness of the hemorrhagic disorder with bleeds being an important cause of mortality. However, with the advent of clotting factor concentrates and the introduction of comprehensive care, a hemorrhagic cause of death became less likely with life expectancy rising to almost that of the non-hemophilic population in the early 1980’s. The onset of the HIV epidemic changed this encouraging statistic, with more than 60% of HIV-infected individuals with hemophilia succumbing to this virus to date. Hepatitis B and C have also taken their toll. However, prior to the HIV epidemic, during the peak period of the epidemic and in the post-HIV period, health care providers continue to express concern about deaths due to suicide. Since 1998 the 135 federally funded Hemophilia Treatment Centers (HTC) have enrolled over 13,000 males with hemophilia, representing over 80% of hemophiliacs who receive care at these HTCs, in the Universal Data Collection (UDC) surveillance program. We used data on all deaths among UDC enrollees, as well as non-UDC enrollees at the HTCs to estimate the incidence of suicide over the nine year period of surveillance. Data from completed mortality forms of both UDC and non-UDC enrollees were analyzed to determine the incidence of suicide episodes for the period 1998 through 2007. There were a total of 17 suicide deaths over this time, producing a crude rate of 12.5/100,000 person years, compared with the rate for the US male population of 17.7/100,000. Because the hemophilia population is younger than the general population, age adjusted suicide rates were also calculated. Detailed age distribution data were available only for persons enrolled in UDC. Therefore, adjusted rates were based on 8 suicides among the 554 deaths that were reported for persons enrolled in UDC. All 8 suicides occurred among White non Hispanic males between the ages of 20 and 61 (Mean =36.1 years). Three of the persons who died from suicide were HIV positive, one was HIV and HCV positive, one HCV and HBV positive, and one was HCV positive. The crude rate of suicide for persons enrolled in UDC was 16.1/100,000 persons years. Using the indirect age adjustment method the adjusted rate is 11.5/100,000 person years, based on 2004 census data, resulting in 5.8 predicted deaths. The observed and expected numbers of deaths were not statistically significantly different. The available data does not indicate a suicide mortality rate higher among hemophilia patients receiving care at HTCs enrolled in UDC than the US male population.
PK Comparability of VWF/FVIII Concentrates after Repeated Dosing
Authors: Kronthaler Ulrich , Stockschlaeder Marcus , Dickneite Gerhard 
Published: April 2020

Abstract
Introduction: Appropriate correction of hemostasis in VWD is essential not only to stop acute perioperative bleeding, but also to promote postoperative wound healing. In order to prevent a drop of the VWF plasma level below the hemostatically necessary concentration, repeated infusions of VWF containing concentrates are required. Plasma derived VWF/FVIII concentrates vary, however, in several aspects such as the VWF/FVIII ratio. As therapy of VWD primarily aims at correcting functional VWF level, the question arises, whether VWF/FVIII concentrates differ with regard to the plasma level of FVIII resulting from repeated concentrate infusions.
Methods: The goal of the present study was to compare VWF and FVIII levels after repeated administration of 150 IU (VWF:RCo)/kg (7 doses at 4-hour intervals). Two plasma derived VWF/FVIII concentrates, Humate-P®/Haemate® P and Wilate®, were selected for the comparison. The concentrates were administered as i.v. bolus to CHB rabbits (n=6/group). Blood samples were collected for close monitoring of the resulting substitution levels. The VWF and FVIII levels were quantified using commercially available ELISA assays.
Results: Pharmacokinetic analysis (AUC, Cmax, half-life, MRT, clearance) demonstrated that the time course of VWF plasma levels were similar. The ratio of the VWF AUC of Humate-P®(Haemate® P)/Wilate® was 0.98 and the maximal VWF plasma concentration of Humate-P®/Haemate® P was slightly higher, at a ratio of 1,16. In contrast, the FVIII levels continuously accumulated upon treatment with Wilate® leading to trough levels, which were about 1,6 fold higher compared to Humate-P®/Haemate® P (Figure). Accordingly, also the peak FVIII level was 1,7 fold higher upon treatment with Wilate®, while the half-life of FVIII did not differ substantially between the two concentrates. The FVIII trough level of Wilate® together with its higher peak level resulted in an AUC, which was 2 fold higher (p&lt;0.0001) than observed for Humate-P®/Haemate® P.
Conclusion: Against the background of Wilate® containing a two fold higher FVIII concentration compared to Humate-P®/Haemate® P, such differential effects appear plausible. At nominally equal VWF:RCof repeated doses, the associated AUC with respect to FVIII:Ag of Humate-P®/Haemate® P was significantly lower, as compared to Wilate®. This is obviously due to the lower FVIII content in the Humate-P®/Haemate® P product. Therefore, a lower accumulation of FVIII trough levels was observed for Humate-P®/Haemate® P as compared to Wilate®.
Figure Figure
Interaction Between Oral Contraceptive Use and Coagulation Factor Levels in Deep Vein Thrombosis
Authors: Zotz Rainer B. , Kloeckner Simone , Scharf Rudiger E. , Gerhardt Andrea 
Published: April 2020

Abstract
Background: Since not all women using oral contraceptives (OC) develop deep vein thrombosis (DVT), the presence of additional risk factors in women with DVT and OC use is likely. High levels of coagulation factor activities are established risk factors of DVT. There are, however, limited data on the combined effect of OC use and high levels of procoagulant coagulation factors, a shortened aPTT or markers of inflammation like erythrocyte sedimentation rate (ESR) or CRP.
Materials and Methods: 246 women with a first DVT (123 associated with OC use) and 246 normal women matched for age and date of birth were evaluated. Cut-off values for continuous variables were the 90% percentile determined in normal women. OC use influences the levels of many coagulation factors. Using the mean effect of OC use on coagulation factor levels (measured in the control subjects), we estimated the coagulation factor levels in patients prior to the index date, i.e. the time of the thrombosis.
Results: Use of OC was associated with an increase in risk of 3.0. This risk increased due to interaction with factor I (FI) (Odds Ratio (OR) 14.2, p=0.0001), FVIII (OR 5.1, p=0.0001), FIX (OR 5.5, p=0.0001), FXI (OR 12.0, p=0.0001), vWF:Ag (OR 7.7, p=0.0001) and aPTT (OR 20.2, p=0.004). In contrast, no significant increase in risk was found when OC use was associated with high levels of ESR or CRP.
Conclusion: The results of this study indicate that the joint effects of OC use and high coagulation factor levels increase the risk of DVT. This risk increase appears to be of clinical relevance particularly in individuals with shortened aPTT values or high levels of fibrinogen.
First Census of Patients with Hemophilia a/B and Von Willebrand Disease Type 3 in the Eastern Part of Germany since 1988
Authors: Koscielny Juergen , Scholz Ute , Klamroth Robert 
Published: April 2020

Abstract
Diagnostics and treatment of severe forms of haemorrhagic diatheses (HD), such as haemophilia A, B or type 3 von Willebrand disease (VWD) need high standards in haemophilia treatment centers (HTCs). Due to their generally low incidence, a close cooperation of haemophilia treaters is needed to optimize treatment strategies. For this purpose, the “Kompetenznetz Hämorrhagische Diathesen Ost” was founded. The first project was the conduction of a survey of epidemiological data of patients with HA, HB and type 3 VWD in all HTCs in the eastern part of Germany. The study assessed the data regarding numbers of patients treated with HA, HB or type 3 VWD, disease severity, regime of coagulation factor replacement (CFR) and the frequency of factor VIII and IX inhibitors. Data of 838 patients (a prevalence of 52.38 per 1 million citizen) from 27 HTCs were evaluated (5–151 per HTCs). Among the included patients, 201 (prevalence: 12.56/million) were children or adolescents (&lt; 18 years). 81 children suffered from severe HA (prevalence: 5.06/million), 20 from severe HB (prevalence: 1.25/million) and 10 from type 3 VWD (prevalence: 0.63/million). In 637 adults (prevalence: 39.82/million) we found 246 with severe HA (prevalence: 15.38/million), 51 severe HB (prevalence: 3.19/million) and 28 VWD of type 3 (prevalence: 1.75/million). The most commonly used treatment of CFR in patients with severe disease was prophylactic modality. 90% of the children received prophylaxis, but also in 64% of the adults an intermittent or longterm secondary prophylaxis was the preferred treatment. At the time of survey conduction, a factor VIII or IX inhibitor was present in 1.9% of the children and in 1.6% of the adults with haemophilia. Our data allow an overview of the number of patients and treatment strategies in the eastern part of Germany.
Complement Deposition Is Predictive of a Low Absolute Immature Platelet Fraction in Immune Thrombocytopenic Purpura
Authors: Andemariam Biree , Bussel James B , Peerschke Ellinor I.B. 
Published: April 2020

Abstract
The role of platelet-associated complement in the pathophysiology of immune thrombocytopenic purpura (ITP) is not well understood. Although various humoral and cell mediated mechanisms have accounted for the decrease in platelet count, the role of the complement system has not been well defined. To test this, we examined complement fixation on platelets using plasma of 100 ITP patients and 50 healthy volunteers. In situ complement fixation was measured as C1q deposition on heterologous platelets using an ELISA approach. Deposition of C1q was evaluated using specific primary monoclonal antibodies (Quidel, San Diego, CA), and an alkaline phosphatase conjugated goat anti mouse secondary antibody and p-NPP substrate to quantify primary antibody binding. Substrate conversion was measured at 405 nm and was proportional to antigen deposition (complement component) on the platelet surface. Controls for specificity included wells coated with blocking solution only, and platelets exposed to buffer instead of plasma. Results for patient samples were normalized relative to an internal assay control (pooled normal plasma) and expressed as a ratio of C1q deposition. Ratios equal to or greater than 1.9 were considered positive for complement fixation. No significant relationship was observed for complement fixation when samples were stratified by gender, splenectomy status, prior treatment or total platelet count. However, a significant relationship between Absolute Immature Platelet Fraction (A-IPF) (Sysmex, Kobe, Japan) and C1q deposition was observed. The data suggest that a positive complement fixation test defined by C1q deposition is predictive of a low A-IPF (&lt;15) with a positive predictive value and specificity of 100%. The two tailed Fisher exact test gives a p-value of 0.0265. In conclusion, these data suggest that complement fixation in the classical pathway may play a role in reducing the number of immature platelets in patients with ITP, perhaps by a direct effect of complement deposition on megakaryocytes and therefore anti-complement therapy may have clinical utility. Further studies that include an examination of complement fixation in the bone marrow, especially on megakaryocytes, may elucidate the exact mechanism of this destruction. Alernatively, flow cytomety of immature platelets to compare them to mature platelets in regard to complement deposition would be of interest.