bluuebunny/crossref_metadata_2025_split huggingface dataset:
MicroRNAs Specific for Immune-Privileged Diffuse Large B-Cell Lymphoma.
Authors: Robertus JanLukas , Harms Geert , Blokzijl Tjasso , de Jong Daphne , Booman Marije , Rosati Stefano , Schuuring Ed , Kluin Philip M. , van den Berg Anke 
Published: October 2019

Abstract
Introduction Primary diffuse large B-cell lymphomas (DLBCL) constitute a heterogeneous group of tumors with a great diversity in both clinical presentation and outcome. Besides nodal DLBCL, about 40% of these tumors present in different extranodal sites, both immune privileged sites such as the testis and the central nervous system (PCNSL) but also non immune privileged sites such as bone, stomach, skin and mediastinum. Remarkably each of these sites show a marked divergence in biological behavior. Recent studies have shown that certain non-coding short RNAs (22nt) called miRNAs may play a role in the pathogenesis of DLBCL. These miRNAs posttrancriptionally down regulate expression of genes. Aim of study To examine the expression levels of selected miRNAs in different DLBCL sites.
Methods and Materials 51 cases of Ann Abour stage I and II primary DLBCL were divided into 4 groups; 19 nodal cases, 11 non-immune privileged site-associated (non-IP) extranodal cases, 9 PCNSL, and 12 testicular DLBCL. Each case was classified GCB or non-GCB using the algorithm as described by Hans et al. 13 miRNAs were selected from current literature on the basis of their involvement in B-cell lymphoma (miR-15, -16, -21, -221, -155, - 181a, -18a, -19, -20a, 17-3p, -17-5p and -92). The expression levels of the mature microRNAs were determined by qRT-PCR using Taqman miRNA assays. Results were compared by using 2−ΔCt and 2−ΔΔCt. Differences in expression levels between the four groups was tested using ANOVA.
Results Of the selected microRNAs, miR-127, miR-17-5p and miR-221 show a significant difference in expression level between the immune privileged site-associated DLBCL (IP-DLBCL) and nodal DLBCL. Both miR-17-5p (p=0.0003) and miR-221 (p=0.0032) show an increased expression level in PCNSL compared to nodal DLBCL and miR-127 shows a significant increased level in testicular DLBCL compared to nodal (p=0.0011) and non-IP (p=0.0393) DLBCL. Also of interest is the fact that miR-17-5p and miR-127 show a significant difference in the level of expression between both PCNSL and testicular DLBCL. MiR-155 shows no significant difference in expression levels between nodal and extranodal DLBCL. Analysis on the basis of GCB and non-GCB classification is currently being undertaken.
Conclusion In PCNSL miR-17-5p and miR-221 are specifically increased whereas miR-127 shows a specifically higher expression in testicular DLBCL when compared to non-IP and nodal DLBCL. Also PCNSL and testicular DLBCL differ in expression of miR-17-5p and miR-127. In contrast to previous reports miR-155 does not show any significant difference between nodal and extranodal groups. This may be caused by the selection of stage I or II primary DLBCL in contrast to previous publications which used a more heterogeneous group of DLBCL.
Intermediate Filaments of Endoskeleton within Human Erythrocytes.
Authors: Terasawa Kazutaka , Taguchi Takehito , Momota Ryusuke , Naito Ichiro , Ohtsuka Aiji 
Published: October 2019

Abstract
The mammalian erythrocytes had been considered to keep their deformable biconcave shape solely by the membrane and the underlying membranous cytoskeleton. However our previous study showed new aspects that human erythrocytes possessed a cytoplasmic endoskeleton apart from the classical membranous cytoskeleton containing beta-actin and neurofilament heavy chain (NF-H) [Terasawa K, et al. Arch. Histol. Cytol2006; 69: 329–340]. Several studies showed that the intermediate filaments such as neurofilament light chain (NF-L), synemin and vimentin were detected in the nucleated erythrocytes [Granger BL, et al. Science.1983; 221: 553–556] and extended from the nuclear envelope to the membranous marginal bands [Granger BL, et al. J. Cell Biol.1982; 92: 299–312]. In the present study, we asked whether the intermediate filaments such as NF-L and vimentin also exist within the newly found endoskeleton of anucleated erythrocytes. NF-L and vimentin were detected within erythrocytes by western blotting and immunofluorescence. The NF-L were detected along the hillside filament group running parallel to the slope between convex periphery and the concave center and vimentin was distributed along several filaments in the whole erythrocytes by the ion-etching/scanning electron microscopy combined with immunocytochemistry. At the hillside region NF-L co-existed with vimentin along the hillside filaments by back scatter emission signals. We concluded that the endoskeletal filaments of the erythrocyte contain intermediate filaments such as NF-L, NF-H and vimentin as those of the nucleated erythrocytes of avian or lower vertebrates. The intermediate filaments of the endoskeleton within the erythrocytes may keep its unique biconcave disc shape against external circumstances.
Pharmacokinetics of Nilotinib in Subjects with Normal or Impaired Hepatic Function.
Authors: Holland Jaymes , Tanaka Chiaki , Zhou Wei , Sethuraman Venkat , Heuman Douglas M. , Schran Horst 
Published: October 2019

Abstract
Nilotinib, a selective and potent inhibitor of Bcr-Abl tyrosine kinase, has demonstrated activity at 400 mg twice daily dosing against imatinib-resistant forms of Bcr-Abl in patients with CML or Ph+ALL in an open label phase I/II study. In this patient population, elevated liver transaminases levels over baseline, which may be related to a combination of past medical history and the hepatotoxic effects of previous medications, are frequently observed but are generally mild and self-limiting. Nilotinib is metabolized in the liver by CYP3A4, and the metabolites are excreted exclusively via the bile into feces. This study was designed to assess pharmacokinetics of nilotinib across a broad spectrum of hepatic impairment in comparison to a control group with normal hepatic function in order to provide guidance for the safe use of the drug in patients with hepatic impairment. Hepatic impairment was categorized using the three Child-Pugh categories (mild, moderate, and severe). The demographics of the impaired and control group were similar with respect to ethnicity, age (years) impaired (47–64) control (48–62), BMI (kg/m2) impaired (23.6–34.7) control (22.8–32.0), and weight (kg) impaired (73.9–103.9) control (73.3–109.5). Serum samples for determination of nilotinib serum concentration by liquid chromatography tandem mass spectrometry were collected up to 120 hours following a single oral 200 mg dose of nilotinib after an overnight fast. Pharmacokinetic parameters were derived from the individual serum concentration-time data by non-compartmental methods. Pre-dose (nilotinib-free) serum samples were obtained to determine ex vivo nilotinib serum protein binding. The results from 14 subjects (6 subjects in each of the mildly and moderately impaired groups and 8 in the control group) in this ongoing study are reported here. Nilotinib exposure at 200 mg dose was similar in patients from a previous clinical study and subjects in the control group of this study. Nilotinib AUC was increased on average by 18% in the mild impairment group, and 23% in the moderate impairment group as compared to the control group. A total of 6 grade 1 and 2 adverse events were reported comprising of back pain, worsening abdominal pain, headache, nausea, heart burn, and indigestion. One adverse event (grade 2 worsening back pain) was reported in the control group. No laboratory or cardiac monitoring findings were considered as adverse events during the treatment period. The currently available results suggest that the pharmacokinetic profile of nilotinib is comparable between subjects with normal hepatic function and subjects with mild or moderate hepatic impairment.
Global Epigenomic Profiling Demonstrates That Myelodysplasia Is Characterized by a Distinct Epigenetic Signature with Aberrant DNA Methylation Changes Involving Various Malignant and Hematopoietic Pathways.
Authors: Zhou L. , Opalinska J. , Sohal D. , Thompson R. , Li Y. , Kotla V. , Pahanish P. , Parmar S. , Kambhampati S. , Figueroa M. , Ye K. , Friedman E. , Wickrema A. , Melnick A. , Greally J. , Verma A. 
Published: October 2019

Abstract
Myelodysplasia (MDS) is a clonal hematopoietic disorder that leads to ineffective hematopoiesis and peripheral cytopenias. DNMT inhibitors such as azacytidine have led to clinical responses in patients, though the genes affected by epigenetic alterations are not well known. Whole genome DNA methylation was analyzed by a recently described novel method, The HELP assay (HpaII tiny fragment Enrichment by Ligation-mediated PCR; Khulan et al, Genome Res. 2006 Aug;16(8)) that uses differential methylation specific restriction digestion by HpaII and MspI followed by amplification, two color labeling and cohybridization to quantitatively determine individual promoter island methylation. A whole genome human promoter array (Nimblegen) was used to determine the level of methylation of 25626 gene promoters by calculating HpaII/MspI cut fragment intensity ratio. Peripheral blood leucocytes from 13 patients with MDS were compared to 9 age matched normal and anemic controls. Gene expression analysis was performed using 37K oligo maskless arrays on cDNA obtained from the same samples. Analysis showed that whole genome methylation profiling has greater discriminatory power in separating clusters of MDS samples from normal and anemic controls when compared to gene expression analysis. Unsupervised clustering based on epigenetic profiling demonstrated that only two cases of early MDS clustered with normals as compared to absolutely no separation between MDS and normals with clustering based on gene expression patterns. A high correlation (r=0.88–0.96) was observed between global methylation profiles of matched sets of bone marrow and peripheral blood leucocyte samples from selected patients demonstrating that peripheral blood leucocytes can be a valid surrogate for epigenomic analysis. Further analysis showed that genes consistently aberrantly methylated in MDS included Syk kinase, HOXB3, several histone acetyltranferases and others. Functional analysis by Ingenuity showed that cancer and cell signaling pathways were the most affected by epigenetic silencing. Most interestingly, a large proportion of gene promoters were also aberrantly hypomethylated. These included genes from Ras oncogene family, the CDC42 GTPase, various methyl binding proteins and other proteins mainly encoding for cancer and hematopoiesis functional pathways, thus biologically validating our analysis. Therefore, our data demonstrates that MDS is characterized by distinct epigenetic aberrations that are preserved in peripheral blood leucocytes. These can be the basis of future studies on pathogenesis and diagnosis for this disease and can potentially uncover a new set of therapeutic gene targets.
Targeting Myeloproliferative Diseases with JAK2 Inhibitors.
Authors: Markovtsov Vadim , Yu Diane , Gelman Marina , Lang Wayne , Taylor Vanessa C. , Huynh Stacey , Frances Roy , Fang Shuling , McLaughlin John , Bhamidipati Sohasekhar , Clough Jeffrey , Singh Rajinder , Park Gary , Sweeney David , Tonkin Elizabeth , Bahjat Rena , Chang Betty , Pine Polly , Daniel Ruby , Payan Donald G. , Holland Sacha , Hitoshi Yasumichi 
Published: October 2019

Abstract
Limited options provided by the current standard of care for the patients suffering from myeloproliferative diseases (MPDs) prompted an extensive search for the underlying molecular mechanisms of these disorders. Recent discovery of a single activating mutation (V617F) in JAK2 kinase gene associated with the development of the polycythemia vera (PV), essential thrombocythemia (ET) and chronic idiopathic myelofibrosis (CIMF) opened up a possibility to develop highly targeted therapies against these debilitating ailments. To that end, we engineered cytokine-independent Ba/F3 cell line expressing the V617F mutant of JAK2 to screen a focused small molecule library for potential inhibitors of JAK2 V617F -dependent proliferation. We confirmed the ability of hit compounds to inhibit proliferation of JAK2-dependent tumor cell lines using UKE-1 and SET-2 cells carrying the V617F JAK2 mutation. A FACS-based phosphoSTAT5 assay was then used to demonstrate that the hits directly targeted mutant JAK2. JAK3 activity of each compound was evaluated in IL-2-dependent CTLL-2 cell line using phosphoSTAT5 FACS and proliferation assays. To avoid hits with nonspecific antiproliferative activity, the hits were tested in JAK2-independent MOLT4, A549 and H1299 cell lines. Compound hits with the desirable properties were further evaluated for their ability to inhibit JAK2, JAK3 and other kinases in the context of T cell, B cell, or mast cell activation using a variety of cell-based assays as well as in the in vitro biochemical assays. We identified a number of compounds that potently inhibit growth of the two V617F mutant cell lines with EC50s varying from 20 to 500 nM, but do not affect proliferation of control cell lines MOLT4, A549 and H1299 to the same degree. These compounds induce strong and highly specific suppression of STAT5 phosphorylation with IC50s of 10 to 200 nM in SET-2 and V617F JAK2 expressing Ba/F3 cells. One of the hits with the desirable biological and pharmacokinetic profiles was further evaluated in V617F JAK2 Ba/F3 engraftment mouse model where it demonstrated significant extension of survival at 150 and 200 mg/kg bid. Such potent JAK2 inhibitors could become the basis for the next generation of compounds targeting JAK2-dependent myeloproliferative diseases.
Pegfilgrastmin in Children with Severe Aplastic Anemia during Immunosuppressive Therapy.
Authors: Pawelec Katarzyna , Matysiak Michal J. 
Published: October 2019

Abstract
Severe neutropenia and infections connected with leukopenia are same of the main complications in immunosuppressive therapy (IST) of severe aplastic anemia (SAA). Granulocyte-colony-stimulating factors are currently given as supportive therapy in SAA protocol treatment beside antithymocyte globulin and cyclosporine A (CSA). We present four cases of girls aged 6–16 years with SAA without family bone marrow donors, who received pegfilgrastmin in IST. The protocol included: antithymocyte globulin (rabbit) 3.75mg/kg/bw iv for 5 days, CSA 5mg/kg/bw orally for 180 days and pegfilgrastmin (Neulasta-Amgen) 6mg in patients over 45kg and 100μg/kg in children less than 45 kg every 10 days sc until absolute neutrophil count (ANC)reached > 1.5x109/l. Blood was monitored every 7 days during therapy. Two patients received 4 doses of pegfilgrastmin. After this therapy the ANC level was normal without any decreases and we observed slow, gradual rise ANC in this girls In these patients there was a complete remission on day 180 of treatment and both are still in remission month 34. The other two patients received 6 doses of pegfilgrastmin. We observed fairly rapid rise and fall of leukocytes during the first 30 days of therapy. The ANC level became normal on day 70 of treatment. One of the patients is now in partial remission day 320. The other girl did not respond to the first course of IST and she received the second course of IST due to the lack of an unrelated bone marrow donor. Pegfilgrastmin was also administered during this therapy. After 3 doses of pegfilgrastmin leukocyte level rose to 5.3x109/l and ANC level to 2.3x109/l. At present the patient is in partial remission and is still receiving CSA day 210. Pegfilgrastimn administered every 10 days was well tolerated by all four patients and there were no adverse events. We observed that pegfilgrastmin was as effective as filgrastmin which was previously administered. No changes were noted during ANC recovery. However administration of pegfilgrastmin reduced the number of injections given and improved the patient’s well-being.
Vorinostat (SAHA) Inhibits STAT6 Phosphorylation and Transcription, Downregulates Bcl-xL, and Induces Apoptosis in Hodgkin Lymphoma (HL) Cell Lines.
Authors: Buglio Daniela , Georgakis Georgios , Arima Kazuhiko , Liu Yong-Jun , Younes Anas 
Published: October 2019

Abstract
Vorinostat (SAHA) Inhibits STAT6 Phosphorylation and Transcription, Downregulates Bcl-xL, and Induces Apoptosis in Hodgkin Lymphoma (HL) Cell Lines. Although the malignant Hodgkin and Reed Sternberg (HRS) cells of HL are of B-cell origin, they infrequently express B-cell antigens. Recent studies demonstrated that several B-cell specific genes are silenced in HRS cells by epigenetic mechanism, suggesting that this process may be reversible and could be explored therapeutically with deacetylase (DAC) inhibitors or hypomethylating agents. Pan-DAC and isotype-selective DAC inhibitors have shown promising activity in vitro and in vivo in a variety of lymphoid malignancies, including HL. However, the mechanisms of antiproliferative action of DAC inhibitors in HL remain unknown. In this study, we examined the antiproliferative effects of the pan-DAC inhibitor vorinostat (inhibits class I and class II DACs) on HL cell lines and determined its effect on signaling mechanisms that are known to promote HRS cell survival, including STAT3, STAT6, Akt, and ERK pathways. Vorinostat inhibited DACs as evident by the increase in histone-3 acetylation as early as 30 minutes of incubation. Furthermore, vorinostat induced the expression of the cell cycle regulatory protein p21 which was associated with an early increase in the G2M cell cycle fraction in HL cells. Vorinostat had no inhibitory effect on SATA3 or ERK, but inhibited STAT6 phosphorylation and transcription in a dose and time dependant manner. This effect was associated with a decrease in Akt phosphorylation on Ser473 residue. Because STAT6 has been reported to transcriptionally regulate Bcl-XL and Thymus and activation-regulated chemokines (TARC), we examined the effect of vorinostat on these two targets in HL cells. TARC has been shown to play an important role in attracting Th2-type T cells and T-regulatory (T-reg) cells, and to be elevated in sera from patients with HL. Vorinostat downregulated the mRNA expression of TARC in a dose dependent manner, suggesting that it may have a role in regulating chemotaxis of reactive T cells and T-reg cells to HL microenvironment in vivo. Moreover, vorinostat reduced the cellular level of the antiapoptotic protein Bcl-xL which was associated with activation of the caspase pathway, and induction of apoptosis in HL cells. Collectively, these data suggest that DAC inhibition in HL by vorinostat may induce cell death by inhibiting STAT6 and downregulating its antiapoptotic target bcl-xL protein. Furthermore, our data suggest that DAC inhibition may have an added effect in vivo on the cellular component in HL microenvironment by inhibiting TARC.
Intravenous Immunoglobulins Regulation of Human B Cells Is Mediated through Extracellular Signal-Regulated Kinase 1 and 2.
Authors: Ducas Éric , Dussault Nathalie , Racine Claudia , Jacques Annie , Paré Isabelle , Côté Serge , Néron Sonia 
Published: October 2019

Abstract
Intravenous immunoglobulin (IVIg) has become increasingly important as therapy in autoimmune and inflammatory diseases, but its immunomodulatory action is still not fully understood. In the treatment of autoimmune diseases, the large repertoire of human immunoglobulin G (IgG), which constitutes IVIg, is assumed to modulate several immune cells including B cells. As a matter of fact, IVIg can act directly upon B cells in vitro by reducing their expansion and enhancing the differentiation of small populations of both naïve and memory B cells. In the present work, we used a panel of human cell lines representative of B cell major maturation stages and treated them with IVIg in order to identify model cell lines representative of normal B cells that could be used to further investigate IVIg effects on B cell physiology and identify the signal transduction pathways involved. Consistent with the observed effects on B cells, one of these cell lines, the DB cells, which express both FcγRIIB receptor (CD32) and surface IgG (B cell receptor [BCR]), showed reduced proliferation and upregulated secretion in presence of IVIg. As revealed by immunobloting and flow cytometry analysis, DB cells response to IVIg was paralleled by increased phosphorylation of ERK1/2, Gab1 as well as Akt, and addition of ERK1/2 activity inhibitors such as PD98059 to cultures of DB cells abolished the negative IVIg effect on proliferation of this cell line. Furthermore, in contrast to the above described IVIg consequence on ERK1/2 phosphorylation, incubation of DB cells in presence of various amounts of aggregated IgG within intramuscular immunoglobulin (IMIg) or IVIg all prevented ERK1/2 up-regulated phosphorylation. Comparable results were obtained using Pfeiffer cells, which are surface IgG+ as DB cells. Considering that CD32 can induce negative signaling when stimulated by aggregates of monomeric IgG present in IVIg, our results with the DB and Pfeiffer model cell lines thus suggest that the direct effect of IVIg on B cells could involve interaction with their BCR.
Coagulation Factor V-Major Survival Predictor in Patients of Viral Hepatitis Induced Acute Liver Failure.
Authors: Gupta Naveen L. , Sharma Subhadra , Saraya Anoop , Mukhopadhyay Asok K. 
Published: October 2019

Abstract
In acute liver failure, variables favoring survival of patients is based on age, interval between onset of jaundice and encephalopathy, coagulation factor level but all these variables were concluded from the studies that were conducted in Western population where etiology most commonly is paracetamol poisoning and metabolic disorders like Wilson’s disease; where as in Indian subcontinent set up major causative factor is viral hepatitis. Thus we exploited the database collected by observational study to investigate major predictors of survival in viral hepatitis induced acute liver failure. Among 40 patients followed for 1 month(7days) or less if died, there were 12 patients who survived. Tha analysis showed that patients who survived had factor V level on day 3 of diagnosis was in the range of 10%–25% and who died had factor V level below 10% (p<0.004). Thus factor V level on day 3 of diagnosis may help in predicting survival in patients of acute liver failure, thus help in triage for the need of orthotopic liver transplantation which is the only definite curative therapy in patients who have poor survival rates.
t test outcome No. of patients mean Standard deviation p value standard error of mean group statistics Interval died 26 30.6538 10.8552 <0.002 2.1289 survived 14 6.1429 3.3249 0.8886 FactorV day1 died 26 7.5769 2.8309 >0.05 0.5552 survived 14 12.0000 4.3677 1.1673 Factor VIII day1 died 26 225.3396 156.0354 >0.05 30.6011 survived 14 293.3556 206.3757 55.1562 FactorV day3 died 26 5.9615 2.0096 <0.004 0.3941 survived 14 12.9286 4.8272 1.2901 FactorVIII day3 died 26 248.2509 185.7102 >0.05 36.4208 survived 14 274.7226 206.5798 55.2108 Lactate day1 died 26 74.3308 11.5141 >0.05 2.2581 survived 14 59.0929 8.0789 2.1592 Lactate day3 died 26 78.9500 8.8363 >0.05 1.7329 survived 14 62.5000 9.4086 2.5146 Factor VIII:V day1 died 26 29.8545 16.6385 >0.05 3.2631 survived 14 22.9066 8.9435 2.3902 FActorVIII:V day3 died 26 42.4612 31.8050 <0.003 6.2375 survived 14 20.0944 8.7368 2.3350 pH day1 died 26 7.1365 0.1331 >0.05 2.611E-02 survived 14 7.1443 0.1302 2.648E-02 pH day3 died 26 7.0300 0.1479 >0.05 2.901E-02 survived 14 7.2286 0.1075 2.872E-02 pCO2 day1 died 26 41.8462 7.6036 >0.05 1.4912 survived 14 40.7857 7.5261 2.0114 pCO2 day3 died 26 46.3077 10.3760 >0.05 2.0349 survived 14 43.2857 5.8498 1.5634 pO2 day1 died 26 87.3462 4.3169 >0.05 0.8466 survived 14 86.0000 4.3677 1.1673 pO2 day3 died 26 89.3462 4.0194 >0.05 0.7883 survived 14 89.5000 3.6742 0.9820