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Quantitative RT-PCR

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https://www.readbyqxmd.com/read/29785002/using-single-molecule-mrna-fluorescent-in-situ-hybridization-rna-fish-to-quantify-mrnas-in-individual-murine-oocytes-and-embryos
#1
Fang Xie, Kelsey A Timme, Jennifer R Wood
Changes in abundance of mRNAs during oocyte growth and maturation and during pre-implantation embryo development have been documented using quantitative real-time RT-PCR (qPCR), microarray analyses, and whole genome sequencing. However, these techniques require amplification of mRNAs, normalization using housekeeping genes, can be biased for abundant transcripts, and/or require large numbers of oocytes and embryos which can be difficult to acquire from mammalian species. We optimized a single molecule RNA fluorescence in situ hybridization (RNA-FISH) protocol, which amplifies fluorescence signal to detect candidate transcripts, for use with individual oocytes and embryos...
May 21, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29784898/pretreatment-with-simvastatin-upregulates-expression-of-bk-2r-and-cd11b-in-the-ischemic-penumbra-of-rats
#2
Jian-Ying Zhang, Qing-Ke Bai, Ying-Dong Zhang
Inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A reductases, collectively known as statins, have been shown to minimize cerebral ischemic events in patients. We assessed the mechanisms of simvastatin pretreatment in preventing cerebral ischemia/reperfusion injury in rats using a model of middle cerebral artery occlusion (MCAO). Rats were pretreated with simvastatin 14 days prior to MCAO induction. At 3, 24, and 48 hours after reperfusion, bradykinin levels in the ischemic penumbra were assayed by ELISA, mRNA levels of bradykinin B2 receptors (BK-2Rs) and CD11b were measured by fluorescent quantitative real-time PCR (RT-PCR), and co-expression of microglia and BK-2Rs was determined by immunofluorescence...
November 1, 2017: Journal of Biomedical Research
https://www.readbyqxmd.com/read/29782830/toxoplasma-gondii-infections-in-chickens-performance-of-various-antibody-detection-techniques-in-serum-and-meat-juice-relative-to-bioassay-and-dna-detection-methods
#3
G Schares, M Koethe, B Bangoura, A-C Geuthner, F Randau, M Ludewig, P Maksimov, M Sens, A Bärwald, F J Conraths, I Villena, D Aubert, M Opsteegh, J Van der Giessen
Chickens, especially if free-range, are frequently exposed to Toxoplasma gondii, and may represent an important reservoir for T. gondii. Poultry products may pose a risk to humans, when consumed undercooked. In addition, chickens are regarded as sensitive indicators for environmental contamination with T. gondii oocysts and have been used as sentinels. The aim of the present study was to determine the suitability of commonly used antibody detection methods, i.e. the modified agglutination test (MAT), IFAT and ELISA to detect T...
May 18, 2018: International Journal for Parasitology
https://www.readbyqxmd.com/read/29781095/gasoline-exhaust-damages-spermatogenesis-through-downregulating-%C3%AE-6-integrin-and-%C3%AE-1-integrin-in-the-rat-model
#4
L Luo, E Li, S Zhao, J Wang, Z Zhu, Y Liu, J Luo, Z Zhao
The health influence of air pollution has been an international public health concern. Increasing evidence has suggested that air pollution has been associated with decreased sperm quality. However, the underlying molecular mechanisms are still not fully elucidated. We aimed to verify whether gasoline exhaust leads to reproductive impairment by injuring spermatogonial stem cells and explore its underlying molecular mechanism. Twenty male Sprague-Dawley rats were randomly divided into two groups: the exposure group (n = 10) and the control group (n = 10)...
May 20, 2018: Andrologia
https://www.readbyqxmd.com/read/29780482/an-exploration-of-nucleic-acid-liquid-biopsy-using-a-glucose-meter
#5
Yu Gu, Ting-Ting Zhang, Zhi-Feng Huang, Shan-Wen Hu, Wei Zhao, Jing-Juan Xu, Hong-Yuan Chen
The development of non-invasive techniques for the diagnosis of cancer, characterization of mutation and monitoring treatment response could greatly reduce the morbidity and mortality caused by cancer. Nevertheless, the extremely low amount of cell free nucleic acids makes liquid biopsy a very challenging task. Herein, taking advantage of the pocket size, reliable quantitative results and simple operation of the pocket-sized personal glucose meter (PGM), we report an approach of circulating microRNA-21 (miR-21) detection with high precision and low cost...
April 14, 2018: Chemical Science
https://www.readbyqxmd.com/read/29780378/selection-and-validation-of-reference-genes-for-quantitative-real-time-pcr-normalization-under-ethanol-stress-conditions-in-oenococcus-oeni-sd-2a
#6
Shuai Peng, Longxiang Liu, Hongyu Zhao, Hua Wang, Hua Li
The powerful Quantitative real-time PCR (RT-qPCR) was widely used to assess gene expression levels, which requires the optimal reference genes used for normalization. Oenococcus oeni ( O. oeni ), as the one of most important microorganisms in wine industry and the most resistant lactic acid bacteria (LAB) species to ethanol, has not been investigated regarding the selection of stable reference genes for RT-qPCR normalization under ethanol stress conditions. In this study, nine candidate reference genes ( proC, dnaG, rpoA, ldhD, ddlA, rrs, gyrA, gyrB , and dpoIII ) were analyzed to determine the most stable reference genes for RT-qPCR in O...
2018: Frontiers in Microbiology
https://www.readbyqxmd.com/read/29780284/up-regulation-of-ctd-2547g23-4-in-hepatocellular-carcinoma-tissues-and-its-prospective-molecular-regulatory-mechanism-a-novel-qrt-pcr-and-bioinformatics-analysis-study
#7
Dong-Yue Wen, Peng Lin, Hai-Wei Liang, Xia Yang, Hai-Yuan Li, Yun He, Hong Yang, Gang Chen
Background: Dysregulated expression of long non-coding RNAs (lncRNAs) has been reported in the pathogenesis and progression of multiple cancers, including hepatocellular carcinoma (HCC). LncRNA CTD-2547G23.4 is a novel lncRNA, and its role in HCC is still unknown. Here, we aimed to clarify the expression pattern and clinical value of CTD-2547G23.4 and to investigate the prospective regulatory mechanism via bioinformatics analysis in HCC. Methods: To identify differentially expressed lncRNAs in HCC, we downloaded RNA-Seq data for HCC and adjacent non-tumour tissues via The Cancer Genome Atlas (TCGA)...
2018: Cancer Cell International
https://www.readbyqxmd.com/read/29778921/goat-boule-isoforms-identification-mrna-expression-in-testis-and-functional-study-and-promoter-methylation-profiles
#8
Xiaoyan Zhang, Shuai Yu, Qing Yang, Ke Wang, Sihuan Zhang, Chuanying Pan, Hailong Yan, Ruihua Dang, Chuzhao Lei, Hong Chen, Xianyong Lan
A conserved gene in meiosis, the Boule gene is involved in meiosis and spermatogenesis. The deletion of this gene in males blocks meiosis and results in infertility. Alternative splicing variants of the Boule gene have been identified in humans, bovines, and bats, but in dairy goats remains unknown. This study was therefore to detect splicing variants of the goat Boule gene and explore their potential roles in meiosis. Three isoforms, denoted as Boule-a, Boule-b, and Boule-c, were identified in the testes of goats using real-time PCR (RT-PCR) and cloning sequencing...
May 12, 2018: Theriogenology
https://www.readbyqxmd.com/read/29778179/recombinant-adenovirus-delivered-soluble-cd163-and-sialoadhesin-receptors-protected-pigs-from-porcine-reproductive-and-respiratory-syndrome-virus-infection
#9
Wenlong Xia, Zhi Wu, Changming Guo, Shanyuan Zhu, Xinyu Zhang, Xiaoli Xia, Huaichang Sun
Porcine reproductive and respiratory syndrome (PRRS) is one of the most important swine diseases affecting pig industry worldwide. Sialoadehesin (Sn) and CD163 are the two specific receptors for PRRSV infection of porcine alveolar macrophages. Our previous study showed that the soluble Sn receptor Sn4D-Fc and soluble CD163 receptor SRCR59-Fc expressed by the two recombinant adenoviral (rAd) vectors have an additive anti-PRRSV effect in vitro. In the present study, rAd-Sn4D-Fc and rAd-SRCR59-Fc were inoculated into pigs, and the efficient expression of Sn4D-Fc and SRCR59-Fc proteins was detected by ELISA...
June 2018: Veterinary Microbiology
https://www.readbyqxmd.com/read/29777949/evaluation-of-skin-or-sweat-characteristic-mrnas-for-inferring-the-human-origin-of-touched-contact-traces
#10
Tomoko Akutsu, Ken Watanabe, Ayari Takamura, Koichi Sakurada
The source of small amounts of touch DNA, which is transferred from the skin to an object when it is handled or touched, could be an issue in the forensic analysis of criminal cases. Here, we performed an extended evaluation of skin- or sweat-characteristic mRNAs to investigate their usability to infer whether an object has been handled or touched by someone. First, we compared the expression levels of candidate genes between skin swabs and other body fluids by quantitative RT-PCR analysis. Among the analyzed genes, corneodesmosin (CDSN), late cornified envelope 1C (LCE1C), filaggrin (FLG), desmocollin 1, and dermcidin were selected for further analysis on the basis of their specificities and sensitivities...
May 14, 2018: Legal Medicine
https://www.readbyqxmd.com/read/29777440/mir-27a-b-regulates-liver-regeneration-by-posttranscriptional-modification-of-tmub1
#11
Xiang Lan, Guangyao Li, Hongming Liu, Hangwei Fu, Ping Chen, Menggang Liu
BACKGROUND: Transmembrane and ubiquitin-like domain-containing 1 protein (Tmub1) negatively regulates liver regeneration. However, whether this regulation involves posttranscriptional modification of Tmub1 expression is unknown. AIM: The aim of the study was to investigate whether microRNA (miR)-27a/b regulates posttranscriptional modification of Tmub1 and cell proliferation during liver regeneration. METHODS: Tmub1 mRNA 3'-untranslated region (UTR) sequences were analyzed using online software...
May 17, 2018: Digestive Diseases and Sciences
https://www.readbyqxmd.com/read/29777335/a-trisubstituted-pyrazole-derivative-reduces-dmba-induced-mammary-tumor-growth-in-rats-by-inhibiting-estrogen-receptor-%C3%AE-expression
#12
Hanumappa Ananda, Kothanahally S Sharath Kumar, Muddenahalli S Sudhanva, Shobith Rangappa, Kanchugarakoppal S Rangappa
Aberrant expression of estrogen receptor alpha (ER-α) is observed in many pathological complications like breast cancer, endometrial cancer, and in osteoporosis. ER-α plays a vital role in the initiation and progression of breast cancer and confers chemo and radioresistance to the cancer cells by upregulating expression of anti-apoptotic proteins. The synthetic pyrazole derivative 3-(1-(4-bromophenyl)-5-phenyl-1H-pyrazol-3-yl)pyridine (compound 5d) displays significant cytotoxicity against mammary carcinoma cells...
May 18, 2018: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/29775875/the-reduced-level-of-growth-factors-in-an-animal-model-of-depression-is-accompanied-by-regulated-necrosis-in-the-frontal-cortex-but-not-in-the-hippocampus
#13
Mateusz Kucharczyk, Anna Kurek, Bartosz Pomierny, Jan Detka, Mariusz Papp, Katarzyna Tota, Bogusława Budziszewska
In the present study, we asked if the different types of stress alter neuronal plasticity markers distinctively in the frontal cortex (FCx) and in the hippocampus (Hp). To do so, we implemented various stress regimens to analyze changes evoked in these rat brain structures. We utilized several molecular techniques, including western blot, ELISA, quantitative RT-PCR, and various biochemical assays, to examine a range of proteins and subjected rats to behavioral tests to evaluate potential maladaptive alterations...
May 14, 2018: Psychoneuroendocrinology
https://www.readbyqxmd.com/read/29774882/-sleep-wake-rhythm-after-extracorporeal-circulation-in-new-zealand-rabbits
#14
Kaiwei Wang, Jiaqiang Zhang
To evaluate the change of sleep-wake rhythm after extracorporeal circulation (ECC) in New Zealand rabbits, and to explore the role of clock genes in sleep-wake rhythm disorder by ECC.
 Methods: A total of 54 New Zealand rabbits were randomly divided into 3 groups: a normal group (Group N), a sham group (Group S) and a model group (Group ECC). Electrocorticogram (ECOG), electroophthalmogram (EOG) and electromyogram (EMG) were respectively recorded by multipurpose EEG recorder, and the sleep-wake rhythm was also recorded...
April 28, 2018: Zhong Nan da Xue Xue Bao. Yi Xue Ban, Journal of Central South University. Medical Sciences
https://www.readbyqxmd.com/read/29774123/the-association-of-human-endogenous-retrovirus-h-long-terminal-repeat-associating-protein-2-hhla2-expression-with-gastric-cancer-prognosis
#15
Masataka Shimonosono, Takaaki Arigami, Shigehiro Yanagita, Daisuke Matsushita, Yasuto Uchikado, Yuko Kijima, Hiroshi Kurahara, Yoshiaki Kita, Shinichiro Mori, Ken Sasaki, Itaru Omoto, Kosei Maemura, Yoshikazu Uenosono, Sumiya Ishigami, Shoji Natsugoe
Currently, immune checkpoint blockade against members of the B7/CD28 family is being used as a new molecular-targeted therapy, in patients with unresectable advanced or recurrent gastric cancer. Although human endogenous retrovirus-H long terminal repeat-associating protein 2 (HHLA2) is a novel molecule of the B7/CD28 family, the clinical impact of its expression remains uncertain in gastric cancer. Consequently, we examined HHLA2 expression in blood specimens from patients with gastric cancer, and investigated the relationship between its expression and clinicopathological factors to assess its potential power as a prognostic blood predictor...
April 24, 2018: Oncotarget
https://www.readbyqxmd.com/read/29773833/transcriptome-and-mirna-analyses-of-the-response-to-corynespora-cassiicola-in-cucumber
#16
Xiangyu Wang, Di Zhang, Na Cui, Yang Yu, Guangchao Yu, Haiyan Fan
Cucumber (Cucumis sativus L.) target leaf spot (TLS), which is caused by the fungus Corynespora cassiicola (C. cassiicola), seriously endangers the production of cucumber. In this assay, we performed comprehensive sequencing of the transcriptome and microRNAs (miRNAs) of a resistant cucumber (Jinyou 38) during C. cassiicola inoculation using the Illumina NextSeq 500 platform. The possible genes related to the response to C. cassiicola were associated with plant hormones, transcription factors, primary metabolism, Ca2+ signaling pathways, secondary metabolism and defense genes...
May 17, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29773662/validation-of-the-16-gene-recurrence-score-in-patients-with-locoregional-high-risk-renal-cell-carcinoma-from-a-phase-3-trial-of-adjuvant-sunitinib
#17
Brian I Rini, Bernard Escudier, Jean-Francois Martini, Ahmed Magheli, Christer Svedman, Margarita Lopatin, Dejan Knezevic, Audrey D Goddard, Phillip G Febbo, Rachel Li, Xun Lin, Olga Valota, Michael Staehler, Robert J Motzer, Alain Ravaud
PURPOSE: Adjuvant sunitinib prolonged disease-free survival (DFS) (hazard ratio [HR] 0.76) in patients with locoregional high-risk renal cell carcinoma (RCC) in the S-TRAC trial (ClinicalTrials.gov NCT00375674). The 16-gene Recurrence Score (RS) assay was previously developed and validated to estimate risk for disease recurrence in patients with RCC post-nephrectomy. This analysis further validated the prognostic value of RS assay in patients from S-TRAC and explored association of RS results with prediction of sunitinib benefit...
May 17, 2018: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/29772440/inhibition-of-nogo-b-promotes-cardiac-hypertrophy-via-endoplasmic-reticulum-stress
#18
Junli Li, Wenchao Wu, Yanguo Xin, Mingyue Zhao, Xiaojing Liu
AIMS: Nogo-B is a key endoplasmic reticulum (ER) protein that regulates ER stress signaling. However, its role in cardiac hypertrophy remains poorly understood. ER stress is interrelated with autophagy in the process of cardiac hypertrophy. Therefore, we aimed to test the hypothesis that both ER stress and autophagy signaling mediate the function of Nogo-B in cardiac hypertrophy. MAIN METHODS: Rat models of transverse aortic constriction (TAC), neonatal rat cardiomyocytes (NRCMs) stimulated with norepinephrine (Ne) and primary cardiac fibroblasts treated with transforming growth factor β1 (TGF-β1) were used in this study...
May 14, 2018: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/29771428/ampk-regulates-energy-metabolism-through-the-sirt1-signaling-pathway-to-improve-myocardial-hypertrophy
#19
H-W Dong, L-F Zhang, S-L Bao
OBJECTIVE: We investigated the correlations of adenosine monophosphate-activated protein kinase (AMPK), Silence information regulator 1 (SIRT1) and energy metabolism with myocardial hypertrophy. MATERIALS AND METHODS: Myocardial hypertrophy experimental model was established via transverse aortic constriction (TAC)-induced myocardial hypertrophy and phenylephrine (PE)-induced hypertrophic myocardial cell culture. After activation of AMPK, the messenger ribonucleic acid (mRNA) expressions in myocardial tissue- and myocardial cell hypertrophy-related genes, atrial natriuretic peptide (ANP) and β-myosin heavy chain (β-MHC), were detected...
May 2018: European Review for Medical and Pharmacological Sciences
https://www.readbyqxmd.com/read/29771423/effect-of-downregulated-lncrna-nbat1-on-the-biological-behavior-of-glioblastoma-cells
#20
J Liu, W-M Wang, X-L Zhang, Q-H Du, H-G Li, Y Zhang
OBJECTIVE: To investigate the expression of lncRNA neuroblastoma associated transcript 1 (NBAT1) in human glioma cell lines and its underlying mechanism. Effect of NBAT1 on biological behaviors of T98 and U87 cells are also explored. PATIENTS AND METHODS: The mRNA expressions of NBAT1 in 48 cases of glioblastoma tissues and 30 cases of normal brain tissues were accessed by Real-time fluorescence quantitative PCR (RT-PCR). The relationship between mRNA expression of NBAT1 and tumor size, malignancy, and prognosis were analyzed...
May 2018: European Review for Medical and Pharmacological Sciences
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