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Quantitative RT-PCR

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https://www.readbyqxmd.com/read/29166265/selection-of-reference-genes-for-quantitative-real-time-rt-pcr-on-gene-expression-in-golden-pompano-trachinotus-ovatus
#1
X J Chen, X Q Zhang, S Huang, Z J Cao, Q W Qin, W T Hu, Y Sun, Y C Zhou
Golden pompano (Trachinotus ovatus) is an important economically fish species. In this study, with an aim to identify reliable reference genes for quantitative real-time PCR (qRT-PCR) in golden pompano, we evaluated the expression stability of eight housekeeping genes in the presence and absence of poly I:C stimulation in eight tissues. The PCR data was analyzed by geNorm and NormFinder algorithms. The results showed that the expression of all the examined genes exhibited tissue-dependent variations. When under normal physiological condition, geNorm and NormFinder identified B2M and 18S as suitable genes...
September 26, 2017: Polish Journal of Veterinary Sciences
https://www.readbyqxmd.com/read/29165998/characterization-of-natural-organic-substances-potentially-hindering-rt-pcr-based-virus-detection-in-large-volumes-of-environmental-water
#2
Akihiko Hata, Manami Inaba, Hiroyuki Katayama, Hiroaki Furumai
Quantitative detection of pathogenic viruses in the environmental water is essential for the assessment of water safety. It is known that some of natural organic substances interfere with virus detection processes, i.e., nucleic acid extraction and reverse transcription-PCR. Such substances are carried over into a sample after virus concentration. In this study, inhibitory substances in coastal water samples were characterized in view of their effects on efficiency of virus detection and property as organic matters...
November 22, 2017: Environmental Science & Technology
https://www.readbyqxmd.com/read/29165592/folate-deficiency-induces-mitotic-aberrations-and-chromosomal-instability-by-compromising-the-spindle-assembly-checkpoint-in-cultured-human-colon-cells
#3
Xihan Guo, Juan Ni, Yuqian Zhu, Tao Zhou, Xiaoling Ma, Jinglun Xue, Xu Wang
Folates comprise the essential B9 vitamin that act as cofactors and cosubstrates in one-carbon metabolism for both biosynthesis and methylation of DNA and RNA. Folate deficiency (FD) has been shown to induce chromosomal instability (CIN), yet the underlying mechanisms are poorly understood. Here, we used human NCM460 colon mucosal cells as a model to investigate the effect of FD on spindle assembly checkpoint (SAC), a cell-cycle regulatory pathway preventing CIN during mitosis. Cells were maintained in medium containing 1...
November 19, 2017: Mutagenesis
https://www.readbyqxmd.com/read/29164291/bacterial-diversity-in-traditional-doogh-in-comparison-to-industrial-doogh
#4
Hamid Reza Sayevand, Farzaneh Bakhtiary, Angelika Pointner, Marlene Remely, Berit Hippe, Hedayat Hosseini, Alexander Haslberger
Forty-four samples of traditional Doogh and yoghurt were collected from 13 regions of 4 provinces in west of Iran (13 area) and analyzed using molecular methods including PCR, denaturing gradient gel electrophoresis (DGGE) of 16S rDNA, and sequencing. Moreover, collected samples as well as samples from industrially Doogh were analyzed with quantitative real-time PCR (RT-PCR). Analyzed 16S rRNA gene sequences of Doogh samples could be allocated to the presence of Lactobacillus spp. The typical yoghurt starter culture bacteria included four different Lactobacillus species with possible probiotic properties, L...
November 21, 2017: Current Microbiology
https://www.readbyqxmd.com/read/29163988/effect-of-mt3-on-retinal-and-choroidal-tgf-%C3%AE-2-and-has2-expressions-in-form-deprivation-myopia-of-guinea-pig
#5
Tao Li, Xiaodong Zhou, Bing Li, Bo Jiang
Purpose: To confirm its dose-dependent effect on form deprivation myopia and evaluate the effect of MT3 at different tissue concentrations on changes in mRNA and protein expression for TGF-β2 and HAS2. Methods: MT3 was intravitreally injected into deprived eyes at two-day intervals. Refraction was measured by streak retinoscopy after cycloplegia. The axial dimensions were measured by A-scan ultrasound. The quantitative RT-PCR and Western blot were used to detect the changes of TGF-β2 and HAS2 expressions in the retina and choroid of guinea pigs...
2017: Journal of Ophthalmology
https://www.readbyqxmd.com/read/29163776/mir-1268b-confers-chemosensitivity-in-breast-cancer-by-targeting-erbb2-mediated-pi3k-akt-pathway
#6
Wen-Jie Zhu, Xu Chen, Ya-Wen Wang, Hai-Ting Liu, Ran-Ran Ma, Peng Gao
Chemoresistance represents a major obstacle to effective therapy for breast cancer. Emerging evidences associated aberrantly expressed miRNAs with tumor development and chemoresistance. MiR-1268b has never been studied in any cancers before, and its roles in mediating tumor progression and drug resistance are still unclear. Selected from miRNA microarray and confirmed by real-time quantitative PCR (RT-qPCR), miR-1268b was found to be significantly upregulated in drug sensitive and ERBB2 negative tissues, as well as in breast cancer patients with low clinical stage...
October 27, 2017: Oncotarget
https://www.readbyqxmd.com/read/29163675/effects-of-different-concentrations-of-type-i-collagen-hydrogel-on-the-growth-and-differentiation-of-chondrocytes
#7
Deshan Hu, Xiuli Shan
The objective of this study was to analyze the effects of type-I collagen hydrogel of different concentrations on the growth and differentiation of rabbit chondrocytes. Articular cartilage from New Zealand white rabbits was harvested and cultured. Second-generation chondrocytes were collected for in vitro culture with 10, 7, and 5 mg/ml type-I collagen hydrogel, respectively (denoted as groups A, B, and C). After in vitro culture for 1 day, chondrocytes were stained with fluorescein diacetate (FDA)/propidium iodide (PI), and cell viability was observed by laser confocal microscopy...
December 2017: Experimental and Therapeutic Medicine
https://www.readbyqxmd.com/read/29163484/tracking-human-immunodeficiency-virus-1-infection-in-the-humanized-drag-mouse-model
#8
Jiae Kim, Kristina K Peachman, Ousman Jobe, Elaine B Morrison, Atef Allam, Linda Jagodzinski, Sofia A Casares, Mangala Rao
Humanized mice are emerging as an alternative model system to well-established non-human primate (NHP) models for studying human immunodeficiency virus (HIV)-1 biology and pathogenesis. Although both NHP and humanized mice have their own strengths and could never truly reflect the complex human immune system and biology, there are several advantages of using the humanized mice in terms of using primary HIV-1 for infection instead of simian immunodeficiency virus or chimera simian/HIV. Several different types of humanized mice have been developed with varying levels of reconstitution of human CD45(+) cells...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/29160508/detection-and-genotyping-of-human-adenovirus-and-sapovirus-in-children-with-acute-gastroenteritis-in-bel%C3%A3-m-par%C3%A3-between-1990-and-1992-first-detection-of-gi-7-and-gv-2-sapoviruses-in-brazil
#9
Larissa Cristina Prado das Neves Costa, Jones Anderson Monteiro Siqueira, Thayara Morais Portal, Edivaldo Costa Sousa, Alexandre da Costa Linhares, Yvone Benchimol Gabbay, Hugo Reis Resque
INTRODUCTION: Acute gastroenteritis (AGE) is one of the most common causes of morbidity and mortality, especially among children from developing countries. Human adenovirus (HAdV) and sapovirus (SaV) are among the agents that cause AGE. The present study aimed to detect and genotype HAdV and SaV in 172 fecal samples from children with AGE, collected during a surveillance study carried out in a low-income community in Belém, Pará, between 1990 and 1992. METHODS: HAdV was detected by nested PCR, using primers Hex1deg/Hex2deg and NeHex3deg/NeHex4deg...
September 2017: Revista da Sociedade Brasileira de Medicina Tropical
https://www.readbyqxmd.com/read/29160014/loss-of-tenascin-x-gene-function-impairs-injury-induced-stromal-angiogenesis-in-mouse-corneas
#10
Takayoshi Sumioka, Hiroki Iwanishi, Yuka Okada, Yuka Nidegawa, Masayasu Miyajima, Ken-Ichi Matsumoto, Shizuya Saika
To determine the contribution by tenascin X (Tnx) gene expression to corneal stromal angiogenesis, the effects were determined of its loss on this response in TNX knockout (KO) mice. In parallel, the effects of such a loss were evaluated on vascular endothelial growth factor (VEGF) and transforming growth factor β1 (TGFβ1) gene and protein expression in fibroblasts and macrophages in cell culture. Histological, immunohistochemical and quantitative RT-PCR changes determined if Tnx gene ablation on angiogenic gene expression, inflammatory cell infiltration and neovascularization induced by central corneal stromal cauterization...
November 21, 2017: Journal of Cellular and Molecular Medicine
https://www.readbyqxmd.com/read/29157804/anti-parkinsonian-effects-of-%C3%AE-amyrin-are-regulated-via-lgg-1-involved-autophagy-pathway-in-caenorhabditis-elegans
#11
Chia-Cheng Wei, Chun-Han Chang, Vivian Hsiu-Chuan Liao
BACKGROUND: Parkinson's disease (PD) is a neurodegenerative disease that is associated with aging and is characterized as a movement disorder. Currently, there is still no complete therapy for PD. In recent years, the identification and characterization of medicinal plants to cure or treat PD has gained increasing scientific interest. PURPOSE: In this study, we investigated a pentacyclic triterpenoid compound, β-amyrin, which is found in many medicinal plants for its anti-Parkinsonian effects, using Caenorhabditis elegans (C...
December 1, 2017: Phytomedicine: International Journal of Phytotherapy and Phytopharmacology
https://www.readbyqxmd.com/read/29156714/investigation-of-factors-affecting-the-efficacy-of-3c23k-a-human-monoclonal-antibody-targeting-misiir
#12
Sarah E Gill, Qing Zhang, Gary L Keeney, William A Cliby, S John Weroha
MISIIR is a potential target for ovarian cancer (OC) therapy due to its tissue-specific pattern of expression. 3C23K is a novel therapeutic monoclonal anti-MISIIR antibody designed to recruit effector cells and promote cell death through ADCC (antibody dependent cell-mediated cytotoxicity). Our objective was to determine the tolerability and efficacy of 3C23K in OC patient-derived xenografts (PDX) and to identify factors affecting efficacy. Quantitative RT-PCR, immunohistochemistry (IHC), and flow cytometry were used to categorize MISIIR expression in established PDX models derived from primary OC patients...
October 17, 2017: Oncotarget
https://www.readbyqxmd.com/read/29155303/therapeutic-vaccine-against-il-1%C3%AE-improved-glucose-control-in-a-mouse-model-of-type-2-diabetes
#13
Yue Zhang, Xiao-Lin Yu, Jun Zha, Li-Zhen Mao, Jia-Qian Chai, Rui-Tian Liu
AIMS: Inflammation is strongly associated with the mechanisms of β-cell failure in type 2 diabetes. Blockade of the key proinflammatory cytokine IL-1β has been implicated as a promising therapeutic strategy for the prevention and treatment of type 2 diabetes. In this study, we developed an IL-1β-targeted therapeutic vaccine consisting of an IL-1β epitope peptide (A1β) and assessed its efficacy on a diabetic KK-A(y) mouse model. MAIN METHODS: KK-A(y) mice were immunized with A1β for three injections at a 2-week interval...
November 16, 2017: Life Sciences
https://www.readbyqxmd.com/read/29154991/the-rice-constans-like-protein-oscol15-suppresses-flowering-by-promoting-ghd7-and-repressing-rid1
#14
Weixun Wu, Yingxin Zhang, Miao Zhang, Xiaodeng Zhan, Xihong Shen, Ping Yu, Daibo Chen, Qunen Liu, Sittipun Sinumporn, Kashif Hussain, Shihua Cheng, Liyong Cao
The photoperiodic flowering pathway is one of the most important regulatory networks controlling flowering time in rice (Oryza sativa L.). Rice is a facultative short-day (SD) plant; flowering is promoted under inductive SD conditions and delayed under non-inductive long-day (LD) conditions. In rice, flowering inhibitor genes play an important role in maintaining the trade-off between reproduction and yield. In this study, we identified a novel floral inhibitor, OsCOL15, which encodes a CONSTANS-like transcription factor...
November 15, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29153700/heparanase-expression-in-blood-is-sensitive-to-monitor-response-to-anticancer-treatment-in-pancreatic-cancer-a-pilot-study
#15
Wujun Wu, Lin Zhao, Yongtian Yu, Haitian Hu, Huaiping Shi, Qian'an Jia, Lixue Du
BACKGROUND: /Objectives: High heparanase level was shown in maliganant tumor; however, whether or not heparanase may serve as a sensitive marker to monitor response to anticancer treatment is still unknown. METHODS: In the pilot study, heparanase mRNA expression in peripheral blood mononuclear cell fraction (PBMC) and activity in plasma and urine were detected by quantitative real time RT-PCR and heparan-degrading enzyme assay in 31 pancreatic cancer patients. RESULTS: Heparanase mRNA and activity in samples from cancer patients were significantly higher than that in healthy donors...
November 13, 2017: Pancreatology: Official Journal of the International Association of Pancreatology (IAP) ... [et Al.]
https://www.readbyqxmd.com/read/29152645/mir-214-promotes-periodontal-ligament-stem-cell-osteoblastic-differentiation-by-modulating-wnt-%C3%AE-%C3%A2-catenin-signaling
#16
Fengdi Cao, Jialin Zhan, Xufeng Chen, Kai Zhang, Renfa Lai, Zhiqiang Feng
The canonical Wnt/β‑catenin signaling is important in the differentiation of human mesenchymal stem cells into osteoblasts. Accumulating evidence suggests that the expression of β‑catenin is, in part, regulated by specific microRNAs (miRNAs). The aim of the present study was to investigate the putative roles of miRNAs in osteoblast differentiation. Polymerase chain reaction (PCR) arrays were used to identify miRNAs that were differentially expressed between differentiated and non‑differentiated periodontal ligament stem cells (PDLSCs), and reverse transcription‑quantitative PCR (RT‑qPCR) was used for validation...
October 19, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/29151979/effects-of-triple-effective-rna-terna-on-the-inhibition-of-hepatocellular-carcinoma-cells
#17
Yuwen Xue, Tiejun Li, Shuyan Liu, York Yuanyuan Zhu, Guilan Wang, Luyu Fu, Li Chen
The occurrence and development of hepatocellular carcinoma (HCC) is a complicate process involved in genetic mutation and epigenetic regulation. Successful HCC therapy needs multi-targets be involved. The aim of this study was to provide a triple effective RNA (teRNA) which composed of the specific siRNAs targeting NET-1 and VEGF and dsRNA activating TLR3, and explored its anti-HCC roles and mechanism. Real-time quantitative PCR (RT-qPCR), Western blot, immunofluorescence staining, MTT, Annexin V-FITC flow cytometry, Transwell and in-vitro Angiogenesis assay were used to measure the cell biological functions and protein expression analysis...
2017: Journal of Cancer
https://www.readbyqxmd.com/read/29151939/clinical-impact-of-micrornas-associated-with-cancer-stem-cells-as-a-prognostic-factor-in-ovarian-carcinoma
#18
So Youn Cha, Yeon Ho Choi, Sohyun Hwang, Ju-Yeon Jeong, Hee Jung An
Background: Ovarian carcinoma is a highly lethal gynecological malignancy due to its frequent relapses and adoption of chemoresistance. To develop new biomarkers for disease progression in ovarian carcinoma, CSCs, which are considered to contribute to disease relapse and metastasis, were isolated from human ovarian carcinoma tissues, and differentially expressed microRNAs (miRNAs) in CSCs were identified and assessed the clinical implication of expression of these miRNAs. Methods: Primary cancer cells derived from human ovarian carcinomas were cultured and spheroid-forming cells (SFCs) were isolated...
2017: Journal of Cancer
https://www.readbyqxmd.com/read/29151156/deep-rnaseq-indicates-protective-mechanisms-of-cold-tolerant-indica-rice-plants-during-early-vegetative-stage
#19
Raul Antonio Sperotto, Artur Teixeira de Araújo Junior, Janete Mariza Adamski, Denise Cargnelutti, Felipe Klein Ricachenevsky, Ben-Hur Neves de Oliveira, Renata Pereira da Cruz, Rinaldo Pires Dos Santos, Leila Picolli da Silva, Janette Palma Fett
Cold-tolerance in rice may be related to increased cellulose deposition in the cell wall, membrane fatty acids unsaturation and differential expression of several newly identified genes. Low temperature exposure during early vegetative stages limits rice plant's growth and development. Most genes previously related to cold tolerance in rice are from the japonica subspecies. To help clarify the mechanisms that regulate cold tolerance in young indica rice plants, comparative transcriptome analysis of 6 h cold-treated (10 °C) leaves from two genotypes, cold-tolerant (CT) and cold-sensitive (CS), was performed...
November 18, 2017: Plant Cell Reports
https://www.readbyqxmd.com/read/29150565/the-role-of-macrophages-in-the-response-to-tnf-inhibition-in-experimental-arthritis
#20
Qi-Quan Huang, Robert Birkett, Renee Doyle, Bo Shi, Elyssa L Roberts, Qinwen Mao, Richard M Pope
The reduction of synovial tissue macrophages is a reliable biomarker for clinical improvement in patients with rheumatoid arthritis (RA), and macrophages are reduced in synovial tissue shortly after initiation of TNF inhibitors. The mechanism for this initial response is unclear. These studies were performed to identify the mechanisms responsible for the initial reduction of macrophages following TNF inhibition, positing that efflux to draining lymph nodes was involved. RA synovial tissue and synovial fluid macrophages expressed CCR7, which was increased in control macrophages following incubation with TNF-α...
November 17, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
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