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Quantitative RT-PCR

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https://www.readbyqxmd.com/read/28432928/a-humanized-mouse-based-hiv-1-viral-outgrowth-assay-with-higher-sensitivity-than-in-vitro-qvoa-in-detecting-latently-infected-cells-from-individuals-on-art-with-undetectable-viral-loads
#1
Paige Charlins, Kimberly Schmitt, Leila Remling-Mulder, Louise E Hogan, Emily Hanhauser, Kristen S Hobbs, Frederick Hecht, Steven G Deeks, Timothy J Henrich, Ramesh Akkina
Assays that can verify full viral eradication are essential in the context of achieving a cure for HIV/AIDS. In vitro quantitative viral out growth assays (qVOA) are currently the gold standard for measuring latent HIV-1 but these assays often fail to detect very low levels of replication-competent virus. Here we investigated an alternative in vivo approach for sensitive viral detection using humanized mice (hmVOA). Peripheral blood CD4+ T cell samples from HIV subjects on stable ART with undetectable viral loads by RT-PCR were first assayed by in vitro qVOA...
April 19, 2017: Virology
https://www.readbyqxmd.com/read/28432059/increased-amplitude-of-inward-rectifier-k-currents-with-advanced-age-in-smooth-muscle-cells-of-murine-superior-epigastric-arteries
#2
Sebastien Hayoz, Jessica Pettis, Vanessa Bradley, Steven S Segal, William F Jackson
Inward rectifier K(+) channels (KIR) may contribute to skeletal muscle blood flow regulation and adapt to advanced age. Using mouse abdominal wall superior epigastric arteries (SEAs) from either young (3-6 months) or old (24-26 months) male C57BL/6 mice, we investigated whether SEA smooth muscle cells (SMCs) express functional KIR, and how aging may affect KIR function. Freshly dissected SEAs were either enzymatically dissociated to isolate SMCs for electrophysiological recording (perforated patch) and mRNA expression, or used intact for pressure myography...
April 21, 2017: American Journal of Physiology. Heart and Circulatory Physiology
https://www.readbyqxmd.com/read/28431583/overexpression-of-mir-584-5p-inhibits-proliferation-and-induces-apoptosis-by-targeting-ww-domain-containing-e3-ubiquitin-protein-ligase-1-in-gastric-cancer
#3
Qing Li, Zheng Li, Song Wei, Weizhi Wang, Zheng Chen, Lei Zhang, Liang Chen, Bowen Li, Guangli Sun, Jianghao Xu, Qiang Li, Lu Wang, Zhipeng Xu, Yiwen Xia, Diancai Zhang, Hao Xu, Zekuan Xu
BACKGROUND: MicroRNAs are endogenously expressed, small non-coding RNAs that modulate gene expression by targeting specific mRNAs, resulting in translational repression or mRNA degradation. Although miR-584-5p has been reported to play a vital role in various malignancies, its role and the molecular mechanisms underlying the effects of miR-584-5p in gastric cancer (GC) remain to be clarified. In this study, we investigated the role of miR-584-5p in GC. METHODS: The expression of miR-584-5p and its specific target gene were determined in human GC specimens and cell lines by microRNA real-time polymerase chain reaction (RT-PCR), quantitative RT-PCR (qRT-PCR) and Western blot...
April 21, 2017: Journal of Experimental & Clinical Cancer Research: CR
https://www.readbyqxmd.com/read/28431355/genome-wide-identification-and-analysis-of-the-populus-trichocarpa-tify-gene-family
#4
Yue Wang, Feng Pan, Danmei Chen, Wenyuan Chu, Huanlong Liu, Yan Xiang
The plant-specific TIFY proteins are widely present in land plants and play the important roles in the regulation of plant stress-responses. In this study, we carried out a bioinformatics analysis of TIFY genes in Populus trichocarpa by determining the phylogenetic relationship, chromosomal location and gene structure and expression profiles analysis under stresses. The 24 TIFY genes were identified and classified into four subfamilies (ZML, JAZ, PPD and TIFY). The 24 TIFY genes were irregularly located on 13 of the 19 chromosomes; ten gene pairs were involved in large-scale interchromosomal segmental duplication events; we identified 17 collinear TIFY gene pairs in the Populus trichocarpa genome...
April 13, 2017: Plant Physiology and Biochemistry: PPB
https://www.readbyqxmd.com/read/28431281/dengue-virus-replicates-and-accumulates-in-aedes-aegypti-salivary-glands
#5
Vincent Raquin, Louis Lambrechts
Dengue virus (DENV) is an RNA virus transmitted among humans by mosquito vectors, mainly Aedes aegypti. DENV transmission requires viral dissemination from the mosquito midgut to the salivary glands. During this process the virus undergoes several population bottlenecks, which are stochastic reductions in population size that restrict intra-host viral genetic diversity and limit the efficiency of natural selection. Despite the implications for virus transmission and evolution, DENV replication in salivary glands has not been directly demonstrated...
April 18, 2017: Virology
https://www.readbyqxmd.com/read/28429668/the-role-of-mitochondria-associated-reactive-oxygen-species-in-the-amyloid-%C3%AE-induced-production-of-angiogenic-factors-by-arpe-19-cells
#6
L Wu, X Tan, L Liang, H Yu, C Wang, D Zhang, A Kijlstra, P Yang
OBJECTIVE: This study aimed to investigate the mechanisms whereby Amyloid-beta (Aβ) induces the production of angiogenic factors by a human retinal pigment epithelial cell line (ARPE-19) cells. METHODS: ARPE-19 cells obtained from the American Type Culture Collection (ATCC) were utilized in this study. The expression level of vascular endothelial growth factor (VEGF), Interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1) and complement activation fragments C3a and C5a were measured by Real-time quantitative PCR (RT-PCR) and Enzyme-linked immunosorbent assay (ELISA)...
March 31, 2017: Current Molecular Medicine
https://www.readbyqxmd.com/read/28429578/soy-isoflavones-enhance-%C3%AE-defensin-synthesis-and-secretion-in-endometrial-epithelial-cells-with-exposure-to-tlr3-agonist-polyinosinic-polycytidylic-acid
#7
Yotesawee Srisomboon, Sutthasinee Poonyachoti, Chatsri Deachapunya
PROBLEM: β-defensins are important innate chemical barriers that protect the endometrium from pathogen invasion. The effects of soy isoflavones, genistein and daidzein, on the expression and secretion of porcine β-defensins (PBD) in endometrial epithelial cells were investigated under normal or poly I:C-stimulated conditions. METHOD OF STUDY: Primary cultured porcine endometrial epithelial (PE) cells were pretreated with genistein or daidzein followed by poly I:C inoculation...
April 21, 2017: American Journal of Reproductive Immunology: AJRI
https://www.readbyqxmd.com/read/28428796/expression-profiling-coupled-with-in-silico-mapping-identifies-candidate-genes-for-reducing-aflatoxin-accumulation-in-maize
#8
Ramesh Dhakal, Chenglin Chai, Ratna Karan, Gary L Windham, William P Williams, Prasanta K Subudhi
Aflatoxin, produced by Aspergillus flavus, is hazardous to health of humans and livestock. The lack of information about large effect QTL for resistance to aflatoxin accumulation is a major obstacle to employ marker-assisted selection for maize improvement. The understanding of resistance mechanisms of the host plant and the associated genes is necessary for improving resistance to A. flavus infection. A suppression subtraction hybridization (SSH) cDNA library was made using the developing kernels of Mp715 (resistant inbred) and B73 (susceptible inbred) and 480 randomly selected cDNA clones were sequenced to identify differentially expressed genes (DEGs) in response to A...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28428792/identification-of-candidate-genes-and-biosynthesis-pathways-related-to-fertility-conversion-by-wheat-ktm3315a-transcriptome-profiling
#9
Jiali Ye, Yang Duan, Gan Hu, Xingxia Geng, Gaoming Zhang, Pengjiao Yan, Zihan Liu, Lingli Zhang, Xiyue Song
The Aegilops kotschyi thermo-sensitive cytoplasmic male sterility (K-TCMS) system may facilitate hybrid wheat (Triticum aestivum L.) seed multiplication and production. The K-TCMS line is completely male sterile during the normal wheat-growing season, whereas its fertility can be restored in a high-temperature environment. To elucidate the molecular mechanisms responsible for male sterility/fertility conversion and candidate genes involved with pollen development in K-TCMS, we employed RNA-seq to sequence the transcriptomes of anthers from K-TCMS line KTM3315A during development under sterile and fertile conditions...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28428534/expression-of-acid-sensing-ion-channels-in-renal-tubular-epithelial-cells-and-their-role-in-patients-with-henoch-sch%C3%A3-nlein-purpura-nephritis
#10
Li-Ping Yuan, Yan Bo, Zhang Qin, Hua Ran, Wang Li, Yu-Fei Li, Gui Ming
BACKGROUND Acid-sensing ion channels (ASICs) are ligand-gated cation channels activated by extracellular protons. However, the role of ASICs in kidney diseases remains uncertain. This study investigated ASICs expression in kidney tissues and their role in the development of Henoch-Schönlein purpura nephritis (HSPN). MATERIAL AND METHODS The expression of ASIC subunits was examined by immunochemical techniques in the kidney tissue from HSPN patients. Acid-induced ASICs expression in cultured renal tubular epithelial cells was determined by quantitative RT-PCR analysis...
April 21, 2017: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
https://www.readbyqxmd.com/read/28427961/evidence-for-enzootic-circulation-of-rift-valley-fever-virus-among-livestock-in-cameroon
#11
M Rissmann, M Eiden, A Wade, R Poueme, S Abdoulkadiri, H Unger, U Ziegler, T Homeier, M H Groschup
Rift Valley fever virus (RVFV) is an arthropod-borne pathogen, causing serious epidemics in Africa and the Arabian Peninsula. In Cameroon serological data indicate the presence of RVFV, but active circulation of RVFV, causing clinical infections has not been proven yet. For this purpose we carried out a serological and molecular study on a total of 1,953 randomly selected serum samples of small ruminants and cattle, which were collected in years 2013 and 2014 in Cameroon. In a first step, sera were screened serologically using a variety of assay formats to reveal RVFV specific antibodies...
April 17, 2017: Acta Tropica
https://www.readbyqxmd.com/read/28427845/development-of-a-micro-neutralization-assay-for-ebolaviruses-using-a-replication-competent-vesicular-stomatitis-hybrid-virus-and-a-quantitative-pcr-readout
#12
Stella S Lee, Kathryn Phy, Keith Peden, Li Sheng-Fowler
Development of vaccines against highly pathogenic viruses that could also be used as agents of bioterrorism is both a public health issue and a national security priority. Methods that can quantify neutralizing antibodies will likely be crucial in demonstrating vaccine effectiveness, as most licensed viral vaccines are effective due to their capacity to elicit neutralizing antibodies. Assays to determine whether antibodies are neutralizing traditionally involve infectious virus, and the assay most commonly used is the plaque-reduction neutralization test (PRNT)...
April 17, 2017: Vaccine
https://www.readbyqxmd.com/read/28427455/comparison-of-methods-for-detecting-asymptomatic-malaria-infections-in-the-china-myanmar-border-area
#13
Yonghong Zhao, Yan Zhao, Yanmin Lv, Fei Liu, Qinghui Wang, Peipei Li, Zhenjun Zhao, Yingjie Liu, Liwang Cui, Qi Fan, Yaming Cao
BACKGROUND: Sensitive methods for detecting asymptomatic malaria infections are essential for identifying potential transmission reservoirs and obtaining an accurate assessment of malaria epidemiology in low-endemicity areas aiming to eliminate malaria. PCR techniques to detect parasite nucleic acids (DNA or RNA) are among the most commonly used molecular methods. However, most of these methods are of low throughput and cannot be used for large-scale molecular epidemiological studies...
April 20, 2017: Malaria Journal
https://www.readbyqxmd.com/read/28427134/-role-of-bone-marrow-tyrosine-kinase-on-chromosome-x-in-the-production-of-pro-inflammatory-cytokines-from-mouse-mononuclear-macrophages-raw264-7-induced-by-endotoxin-lipopolysaccharide-and-its-mechanism
#14
X Fang, Y Hu, Y Wang, S Liu, F Wang, X L Chen
Objective: To investigate the role of bone marrow tyrosine kinase on chromosome X (BMX) in the production of tumor necrosis factor α (TNF-α) and interleukin-1β (IL-1β) from mouse mononuclear-macrophages induced by endotoxin/lipopolysaccharide (LPS) and its related mechanism. Methods: Mouse mononuclear-macrophages RAW264.7 were inoculated in 6-well plates and routinely cultured for the following experiments. (1) Cells were collected and divided into blank control group, LPS control group, and 75, 750, 7 500, 75 000 nmol/L BMX-IN-1 pretreatment groups according to the random number table, with 8 wells in each group...
April 20, 2017: Zhonghua Shao Shang za Zhi, Zhonghua Shaoshang Zazhi, Chinese Journal of Burns
https://www.readbyqxmd.com/read/28426690/phenotypic-characterization-and-anticancer-capacity-of-cd8-cytokine-induced-killer-cells-after-antigen-induced-expansion
#15
Jianhua Liu, Lu Wang, Yaoling Wang, Wenjie Zhang, Yilin Cao
Cytokine-induced killer cells (CIK) have been used in clinic for adoptive immunotherapy in a variety of malignant tumors and have improved the prognosis of cancer patients. However, there are individual differences in the CIK cell preparations including the obvious differences in the ratio of effector CIK cells among different cancer patients. Infusion of such heterogeneous immune cell preparation is an important factor that would affect the therapeutic efficacy. We report here the enrichment and expansion of CD8+ cells from CIK cells cultured for one week using magnetic activated cell sorting (MACS)...
2017: PloS One
https://www.readbyqxmd.com/read/28425327/a-cross-sectional-quantitative-rt-pcr-study-of-feline-coronavirus-viremia-and-replication-in-peripheral-blood-of-healthy-shelter-cats-in-southern-california
#16
Eric J Fish, Pedro Paulo Vp Diniz, Yen-Chen Juan, Frank Bossong, Ellen W Collisson, Yvonne Drechsler, Bernhard Kaltenboeck
Objectives The objectives of this study were to determine the prevalence of feline coronavirus (FCoV) viremia, and its replication in peripheral blood using quantitative RT-PCR (qRT-PCR) methodology in a population of 205 healthy shelter cats in Southern California, as well as to assess any possible connection to longitudinal development of feline infectious peritonitis (FIP). Methods The study was performed on buffy-coat samples from EDTA-anticoagulated whole blood samples of 205 healthy shelter cats. From 50 of these cats, fecal samples were also examined...
April 1, 2017: Journal of Feline Medicine and Surgery
https://www.readbyqxmd.com/read/28423543/mir-519d-impedes-cisplatin-resistance-in-breast-cancer-stem-cells-by-down-regulating-the-expression-of-mcl-1
#17
Qing Xie, Shuai Wang, Yue Zhao, Zhenchao Zhang, Chuan Qin, Xianjun Yang
Cancer stem cells are considered as the cell population which is responsible for chemoresistance and treatment failure in breast cancer patients. Therefore, it is urgent to explore the mechanism by which cancer stem cells survive under the treatment of chemotherapeutic drugs such as cisplatin. In this paper, we demonstrated significant decrease of miR-519d in breast cancer stem cells by quantitative RT-PCR analysis. Furthermore, we found the enforced expression of miR-519d in T-47D-cancer stem cells significantly increased their sensitivity to cisplatin through the apoptosis pathway...
March 28, 2017: Oncotarget
https://www.readbyqxmd.com/read/28423200/cd80cd86-deficiency-disrupts-regulatory-cd4-foxp3-t-cell-homeostasis-and-induces-autoimmune-like-alopecia
#18
Ji-Hyun Bae, Woo-Sung Hwang, Yoo-Jin Jang, Yoon-Hoo Lee, Da-Eun Jang, Jung-Sik Kim, Su-Cheong Yeom
Alopecia areata (AA) is an autoimmune disease that results in spot baldness in humans. Adequate animal models for AA are currently lacking. The objective of this study was to elucidate the mechanism of autoimmune-like alopecia (ALA) in C57BL/6.CD80CD86-deficient (B6.CD80CD86(-/-) ) mice. Incidence and severity of alopecia were analyzed in 58 B6.CD80CD86(-/-) mice by using histological examination, flow cytometry, multiplex enzyme-linked immunosorbent assay, quantitative RT-PCR, and CD25 inhibition test. Both male and female B6...
April 19, 2017: Experimental Dermatology
https://www.readbyqxmd.com/read/28422864/ketamine-as-adjuvant-analgesics-for-patients-with-refractory-cancer-pain-does-affect-il-2-ifn-%C3%AE-expression-of-t-cells-in-vitro-a-prospective-randomized-double-blind-study
#19
Naibao Zhou, Zhijian Fu, Hao Li, Kaiguo Wang
BACKGROUND: Ketamine has been used as an analgesic adjuvant with morphine in the treatment of refractory cancer pain recently. But both morphine and ketamine have been reported to produce a number of immunomodulatory effects. The current study was performed to assess whether the concentration of ketamine, as adjuvant analgesics for patient with refractory cancer pain, was related to its effect on T cells interleukin-2 (IL-2)/interferon-γ (IFN-γ) expression in vitro. METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated from venous blood of patients with refractory cancer pain over a Ficoll-Hypaque density gradient...
April 2017: Medicine (Baltimore)
https://www.readbyqxmd.com/read/28422321/aberrantly-expressed-long-non-coding-rnas-in-cd8-t-cells-response-to-active-tuberculosis
#20
Yurong Fu, Kunshan Gao, Enxue Tao, Ruifang Li, Zhengjun Yi
Dysregulated expression of long noncoding RNAs (lncRNAs) has been demonstrated as being implicated in a variety of human diseases. In the study we aimed to determine lncRNA profile in CD8(+) T cells response to active tuberculosis (TB). We examined the lncRNA expression by microarray in circulating CD8(+) T cells isolated from patients with active TB and healthy controls. Change predictions to analysis was used to address functional roles of the deregulated mRNAs. Real-time quantitative PCR (RT-qPCR) was used to validate the microarray result...
April 19, 2017: Journal of Cellular Biochemistry
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