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https://www.readbyqxmd.com/read/29342112/useful-bicistronic-reporter-system-for-studying-poly-a-site-defining-cis-elements-and-regulation-of-alternative-polyadenylation
#1
Zhongyuan Deng, Shen Zhang, Shaohua Gu, Xinzhi Ni, Wenxian Zeng, Xianchun Li
The link between polyadenylation (pA) and various biological, behavioral, and pathological events of eukaryotes underlines the need to develop in vivo polyadenylation assay methods for characterization of the cis-acting elements, trans-acting factors and environmental stimuli that affect polyadenylation efficiency and/or relative usage of two alternative polyadenylation (APA) sites. The current protein-based CAT or luciferase reporter systems can measure the polyadenylation efficiency of a single pA site or candidate cis element but not the choice of two APA sites...
January 17, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29336467/nox4-ros-mediate-ethanol%C3%A2-induced-apoptosis-via-mapk-signal-pathway-in-l%C3%A2-02-cells
#2
Cheng-Fang Yang, Yu-Juan Zhong, Zuheng Ma, Li Li, Lin Shi, Li Chen, Chen Li, Dan Wu, Qi Chen, Yong-Wen Li
The aim of the present study was to assess the molecular mechanism of ethanol‑induced oxidative stress‑mediated apoptosis in L‑02 liver cells in order to elucidate novel pathways associated with alcoholic liver disease. L‑02 cells were treated with 400 mM ethanol with or without inhibitors. The cell viability was measured by an MTT assay. Cell apoptosis was assessed by flow cytometry and a single‑stranded DNA (ssDNA) assay. Intracellular reactive oxygen species (ROS) production of L‑02 cells was determined using the 2',7'‑dichlorofluorescein‑diacetate dye...
January 16, 2018: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/29335768/ctla-4-polymorphisms-influence-on-transplant-related-mortality-and-survival-in-children-undergoing-allogeneic-hematopoietic-stem-cell-transplantation
#3
Judith Hammrich, Susan Wittig, Thomas Ernst, Bernd Gruhn
PURPOSE: Allogeneic hematopoietic stem cell transplantation (HSCT) is a curative approach for a variety of hematological diseases; however, it is still associated with substantial morbidity and mortality. Transplant-related mortality (TRM) after HSCT depends mainly on the toxicity of the conditioning regimen, infections, and graft-versus-host disease. The purpose of this study was to identify the association between CTLA-4 single nucleotide polymorphisms and TRM in children undergoing allogeneic HSCT...
January 15, 2018: Journal of Cancer Research and Clinical Oncology
https://www.readbyqxmd.com/read/29335378/comparison-of-telomere-length-measurement-methods
#4
REVIEW
Tsung-Po Lai, Woodring E Wright, Jerry W Shay
The strengths and limitations of the major methods developed to measure telomere lengths (TLs) in cells and tissues are presented in this review. These include Q-PCR (Quantitative Polymerase Chain Reaction), TRF (Terminal Restriction Fragment) analysis, a variety of Q-FISH (Quantitative Fluorescence In Situ Hybridization) methods, STELA (Single TElomere Length Analysis) and TeSLA (Telomere Shortest Length Assay). For each method, we will cover information about validation studies, including reproducibility in independent laboratories, accuracy, reliability and sensitivity for measuring not only the average but also the shortest telomeres...
March 5, 2018: Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
https://www.readbyqxmd.com/read/29335317/reduced-n-type-ca2-channels-in-atrioventricular-ganglion-neurons-are-involved-in-ventricular-arrhythmogenesis
#5
Dongze Zhang, Huiyin Tu, Liang Cao, Hong Zheng, Robert L Muelleman, Michael C Wadman, Yu-Long Li
BACKGROUND: Attenuated cardiac vagal activity is associated with ventricular arrhythmogenesis and related mortality in patients with chronic heart failure. Our recent study has shown that expression of N-type Ca2+ channel α-subunits (Cav2.2-α) and N-type Ca2+ currents are reduced in intracardiac ganglion neurons from rats with chronic heart failure. Rat intracardiac ganglia are divided into the atrioventricular ganglion (AVG) and sinoatrial ganglion. Ventricular myocardium receives projection of neuronal terminals only from the AVG...
January 15, 2018: Journal of the American Heart Association
https://www.readbyqxmd.com/read/29334548/plasma-and-intracellular-pharmacokinetics-of-tenofovir-in-patients-switched-from-tenofovir-disoproxil-fumarate-to-tenofovir-alafenamide
#6
Anthony T Podany, Sara H Bares, Joshua Havens, Ravi Dyavar Shetty, Jennifer O'neill, Sarah Lee, Courtney V Fletcher, Susan Swindells, Kimberly K Scarsi
OBJECTIVES: The aim of the study was to compare the intra-individual plasma and intracellular peripheral blood mononuclear cell (PBMC) pharmacokinetics (PK) of tenofovir (TFV) and its intracellular metabolite, tenofovir-diphosphate (TFV-DP) in patients switched from a fixed dose combination (FDC) tablet of tenofovir disoproxil fumarate (TDF)/ emtricitabine (FTC)/ elvitegravir (EVG)/ cobicistat (COBI) to a FDC containing tenofovir alafenamide (TAF)/FTC/EVG/COBI. DESIGN: A single arm, prospective, non-randomized, cross-over, PK study in patients receiving a TDF-containing regimen (TDF 300 mg/ FTC 200 mg/ EVG 150 mg/ COBI 150 mg) switched to a TAF-containing FDC regimen (TAF 10 mg/ FTC 200 mg/ EVG 150 mg/ COBI 150 mg)...
January 13, 2018: AIDS
https://www.readbyqxmd.com/read/29332296/plant-cell-cultures-as-model-systems-to-study-programmed-cell-death
#7
Sara Cimini, Maria Beatrice Ronci, Elisabetta Barizza, Maria Concetta de Pinto, Vittoria Locato, Fiorella Lo Schiavo, Laura De Gara
The study of programmed cell death (PCD) activated in a certain group of cells is complex when analyzed in the whole plant. Plant cell suspension cultures are useful when investigating PCD triggered by environmental and developmental stimuli. Due to their homogeneity and the possibility to synchronize their responses induced by external stimuli, these cultures are used for studying the signaling pathways leading to PCD. The first problem in the analysis of PCD in cell cultures is the quantification of cell viability/death over time...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29330273/rheumatoid-disease-an-unusual-cause-of-relapsing-meningoencephalitis
#8
Sian K Alexander, Maria Di Cicco, Ute Pohl, Alberto Cifelli
A 73-year-old man presented with three episodes of dysphasia and disinhibited behaviour, a single seizure and transient ischaemic attack-like events characterised by right arm and/or leg weakness. These episodes were separated by month-long asymptomatic intervals. Medical history included rheumatoid arthritis, which was clinically quiescent on leflunomide.Repeated cerebrospinal fluid examination showed a persistent lymphocytosis with mildly reduced glucose and elevated protein; oligoclonal bands and viral PCR were negative...
January 12, 2018: BMJ Case Reports
https://www.readbyqxmd.com/read/29329703/highly-sensitive-microrna-detection-based-on-bio-bar-code-assay-and-catalytic-hairpin-assembly-two-stage-amplification
#9
Songsong Tang, Yuan Gu, Huiting Lu, Haifeng Dong, Kai Zhang, Wenhao Dai, Xiangdan Meng, Fan Yang, Xueji Zhang
Herein, a highly-sensitive microRNA (miRNA) detection strategy was developed by combining bio-bar-code assay (BBA) with catalytic hairpin assembly (CHA). In the proposed system, two nanoprobes of magnetic nanoparticles functionalized with DNA probes (MNPs-DNA) and gold nanoparticles with numerous barcode DNA (AuNPs-DNA) were designed. In the presence of target miRNA, the MNP-DNA and AuNP-DNA hybridized with target miRNA to form a "sandwich" structure. After "sandwich" structures were separated from the solution by the magnetic field and dehybridized by high temperature, the barcode DNA sequences were released by dissolving AuNPs...
April 3, 2018: Analytica Chimica Acta
https://www.readbyqxmd.com/read/29325035/amplicon-based-next-generation-sequencing-of-plasma-cell-free-dna-for-detection-of-driver-and-resistance-mutations-in-advanced-non-small-cell-lung-cancer
#10
N Guibert, Y Hu, N Feeney, Y Kuang, V Plagnol, G Jones, K Howarth, J F Beeler, C P Paweletz, G R Oxnard
Background: Genomic analysis of plasma cell-free DNA is transforming lung cancer care, however available assays are limited by cost, turnaround time, and imperfect accuracy. Here we study amplicon-based plasma next-generation sequencing (NGS), rather than hybrid-capture-based plasma NGS, hypothesizing this would allow sensitive detection and monitoring of driver and resistance mutations in advanced non-small cell lung cancer (NSCLC). Methods: Plasma samples from patients with NSCLC and a known targetable genotype (EGFR, ALK/ROS1 and other rare genotypes) were collected while on therapy and analyzed, blinded to tumor genotype...
January 9, 2018: Annals of Oncology: Official Journal of the European Society for Medical Oncology
https://www.readbyqxmd.com/read/29317874/molecular-cloning-expression-and-characterization-of-plasmid-encoding-rhomboid-4-rom4-of-tachyzoite-of-toxoplasma-gondii-rh-strain
#11
Mohammad Taghi Rahimi, Shahabeddin Sarvi, Mahdi Sharif, Saeid Abediankenari, Ehsan Ahmadpour, Reza Valadan, Ahmad Daryani
Background: The objective of this study was to clone, express and characterize the gene encoding rhomboid 4 (ROM4) proteins, a vital gene in surface adhesion and host cell invasion process of tachyzoite of T. gondii in an appropriate expression vector and eukaryotic cell for production of recombinant protein. Methods: Toxoplasma RNA was isolated from tachyzoites (RH strain) and complementary DNA was synthesized. Oligonucleotide primer pair was designed based on Toxoplasma ROM4 gene sequence with XhoI and EcoRI restriction sites at 5' end of forward and reverse primers, respectively...
October 2017: Iranian Journal of Parasitology
https://www.readbyqxmd.com/read/29317590/the-association-between-variants-of-receptor-for-advanced-glycation-end-products-rage-gene-polymorphisms-and-age-related-macular-degeneration
#12
Mantas Banevicius, Alvita Vilkeviciute, Loresa Kriauciuniene, Rasa Liutkeviciene, Vytenis Pranas Deltuva
BACKGROUND Age-related macular degeneration (AMD) is the leading cause of blindness in people aged 65 years and older in developed countries. The pathogenesis of AMD has been linked to mechanisms involving inflammation, oxidative stress, and basal laminar deposit formation between retinal pigment epithelium (RPE) cells and the basal membrane, caused by advanced glycation end products (AGEs). AGEs are implicated in the pathogenesis of AMD through the AGE-and receptor for AGE (RAGE) interaction, which can be altered by polymorphisms of the RAGE gene...
January 10, 2018: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
https://www.readbyqxmd.com/read/29316067/deep-sequencing-shows-low-level-oncogenic-hepatitis-b-virus-variants-persisting-post-liver-transplant-despite-potent-anti-hbv-prophylaxis
#13
Keith Ck Lau, Carla Osiowy, Elizabeth Giles, Beth Lusina, Guido van Marle, Kelly W Burak, Carla S Coffin
Recent studies suggest that withdrawal of hepatitis B immune globulin (HBIG) and nucleos(t)ide analogues (NA) prophylaxis may be considered in HBV surface antigen (HBsAg) negative liver transplant (LT) recipients with a low risk of disease recurrence. However, the frequency of occult HBV infection (OBI) and HBV variants after LT in the current era of potent NA therapy is unknown. 12 LT recipients on prophylaxis were tested in matched plasma and peripheral blood mononuclear cells (PBMC) for HBV quasispecies by in-house nested PCR and next generation sequencing of amplicons...
January 6, 2018: Journal of Viral Hepatitis
https://www.readbyqxmd.com/read/29315647/antiviral-activity-of-a771726-the-active-metabolite-of-leflunomide-against-jun%C3%A3-n-virus
#14
Claudia S Sepúlveda, Cybele C García, Elsa B Damonte
The aim of this study was to investigate the effect of A771726, the active metabolite of leflunomide, against the infection with Junín virus (JUNV), agent of Argentine hemorrhagic fever (AHF). The treatment with non-cytotoxic concentrations of A771726 of Vero and A549 cells infected with JUNV inhibited virus replication in a dose-dependent manner, as determined by virus yield reduction assay. The antiviral effectiveness of A771726 was not importantly affected by the multiplicity of infection and the virus strain...
January 5, 2018: Journal of Medical Virology
https://www.readbyqxmd.com/read/29314149/uniplex-e6-e7-pcr-method-detecting-e6-or-e7-genes-in-39-human-papillomavirus-types
#15
Mitsuaki Okodo, Kaori Okayama, Koji Teruya, Toshiyuki Sasagawa
We have developed a new human papillomavirus (HPV) assay using the uniplex E6/E7 polymerase chain reaction (PCR) method, which is able to detect E6 and E7 genes in 39 HPV types. We validated the assay for sensitivity and specificity using cloned HPV DNA and clinical samples. A comparative study using Genosearch-31 (GS-31) to determine HPV genotypes in clinical samples was also performed. E6 or E7 genes, measured by uniplex E6/E7 PCR, were detectable in 15 low-risk (HPV-6, -11, -40, -42, -44, -54, -55, -61, -62, -71, -74, -81, -84, -89, -90), 11 intermediate-risk (HPV-26, -30, -34, -53, -66, -67, -69, -70, -73, -82, -85), and 13 high-risk (HR) HPV types...
January 4, 2018: Journal of Medical Virology
https://www.readbyqxmd.com/read/29313979/whole-blood-fpr1-mrna-expression-predicts-both-non-small-cell-and-small-cell-lung-cancer
#16
Scott Morris, Anil Vachani, Harvey I Pass, William N Rom, Kirk Ryden, Glen J Weiss, D K Hogarth, George Runger, Donald Richards, Troy Shelton, David W Mallery
While long-term survival rates for early-stage lung cancer are high, most cases are diagnosed in later stages that can negatively impact survival rates. We aim to design a simple, single biomarker blood test for early-stage lung cancer that is robust to preclinical variables and can be readily implemented in the clinic. Whole blood was collected in PAXgene tubes from a training set of 29 patients, and a validation set of 260 patients, of which samples from 58 patients were prospectively collected in a clinical trial specifically for this study...
January 5, 2018: International Journal of Cancer. Journal International du Cancer
https://www.readbyqxmd.com/read/29306398/intraspecific-bloom-succession-in-the-harmful-dinoflagellate-cochlodinium-polykrikoides-dinophyceae-extended-the-blooming-period-in-korean-coastal-waters-in-2009
#17
Bum Soo Park, Jin Ho Kim, Joo-Hwan Kim, Seung Ho Baek, Myung-Soo Han
Although there have been extensive studies on dinoflagellate blooms in recent decades, the mechanism that allows the maintenance of blooms over long periods remains uncertain, and studies on genetically differentiated subpopulations may provide insights into this mechanism. In this study, the influence of two genetically distinct subpopulations of the dinoflagellate Cochlodinium polykrikoides, referred to as Group I and IV, on bloom duration in Korean coastal waters (KCW) was examined using a quantitative PCR (qPCR) assay...
January 2018: Harmful Algae
https://www.readbyqxmd.com/read/29306397/pentaplacodinium-saltonense-gen-et-sp-nov-dinophyceae-and-its-relationship-to-the-cyst-defined-genus-operculodinium-and-yessotoxin-producing-protoceratium-reticulatum
#18
Kenneth Neil Mertens, M Consuelo Carbonell-Moore, Vera Pospelova, Martin J Head, Andrea Highfield, Declan Schroeder, Haifeng Gu, Karl B Andree, Margarita Fernandez, Aika Yamaguchi, Yoshihito Takano, Kazumi Matsuoka, Elisabeth Nézan, Gwenael Bilien, Yuri Okolodkov, Kazuhiko Koike, Mona Hoppenrath, Maya Pfaff, Grant Pitcher, Abdulrahman Al-Muftah, André Rochon, Po Teen Lim, Chui Pin Leaw, Zhen Fei Lim, Marianne Ellegaard
Strains of a dinoflagellate from the Salton Sea, previously identified as Protoceratium reticulatum and yessotoxin producing, have been reexamined morphologically and genetically and Pentaplacodinium saltonense n. gen. et sp. is erected to accommodate this species. Pentaplacodinium saltonense differs from Protoceratium reticulatum (Claparède et Lachmann 1859) Bütschli 1885 in the number of precingular plates (five vs. six), cingular displacement (two widths vs. one), and distinct cyst morphology. Incubation experiments (excystment and encystment) show that the resting cyst of Pentaplacodinium saltonense is morphologically most similar to the cyst-defined species Operculodinium israelianum (Rossignol, 1962) Wall (1967) and O...
January 2018: Harmful Algae
https://www.readbyqxmd.com/read/29306331/fc-gamma-receptors-are-expressed-in-the-developing-rat-brain-and-activate-downstream-signaling-molecules-upon-cross-linking-with-immune-complex
#19
Marianna Stamou, Ana Cristina Grodzki, Marc van Oostrum, Bernd Wollscheid, Pamela J Lein
BACKGROUND: Exposure of the developing brain to immune mediators, including antibodies, is postulated to increase risk for neurodevelopmental disorders and neurodegenerative disease. It has been suggested that immunoglobulin G-immune complexes (IgG-IC) activate Fc gamma receptors (FcγR) expressed on neurons to modify signaling events in these cells. However, testing this hypothesis is hindered by a paucity of data regarding neuronal FcγR expression and function. METHODS: FcγR transcript expression in the hippocampus, cortex, and cerebellum of neonatal male and female rats was investigated ex vivo and in mixed cultures of primary hippocampal and cortical neurons and astrocytes using quantitative PCR analyses...
January 6, 2018: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/29305710/ecto-5-nucleotidase-cd73-contributes-to-the-radiosensitivity-of-t24-human-bladder-cancer-cell-line
#20
Fabrícia Dietrich, Fabrício Figueiró, Eduardo Cremonese Filippi-Chiela, Angélica Regina Cappellari, Liliana Rockenbach, Alain Tremblay, Patrícia Boni de Paula, Rafael Roesler, Aroldo Braga Filho, Jean Sévigny, Fernanda Bueno Morrone, Ana Maria Oliveira Battastini
PURPOSE: Trimodal therapy is a reasonable bladder-preserving option to radical cystectomy. However, many tumors are radioresistive. In this sense, the identification of new prognostic and predictive biomarkers that allow the selection of patients with better responses to radiation therapy would improve outcomes. With the aim of using ecto-5'-nucleotidase/CD73 as a predictive biomarker, the role of this enzyme in the context of radiotherapy in T24 human bladder cancer cell line was investigated...
January 5, 2018: Journal of Cancer Research and Clinical Oncology
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