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Quantitative RT-PCR

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https://www.readbyqxmd.com/read/29356168/altered-transcription-of-glutamatergic-and-glycinergic-receptors-in-spinal-cord-dorsal-horn-following-spinal-cord-transection-is-minimally-affected-by-passive-exercise-of-the-hindlimbs
#1
Jeremy W Chopek, Christopher W MacDonell, Patricia C Shepard, Kalan R Gardiner, Phillip F Gardiner
Gene expression is altered following a spinal transection (STx) in both motor and sensory systems. Exercise has been shown to influence gene expression in both systems post-STx. Gene expression alterations have also been shown in the dorsal root ganglia and nociceptive laminae of the spinal cord following either an incomplete spinal cord injury (SCI) or a contusive SCI. However, the effect of STx and exercise on gene expression in spinal cord laminae I-III has not fully been examined. Therefore, the purpose of this study was to determine if gene expression in laminae I-III is altered following STx and determine if superimposed passive exercise of the hindlimbs would influence gene expression post-STx in laminae I-III...
January 22, 2018: European Journal of Neuroscience
https://www.readbyqxmd.com/read/29356044/expression-analysis-of-high-mobility-group-box-1-hmgb1-in-histologic-samples-of-malignant-pleural-mesothelioma
#2
Eltjona Rrapaj, Elena Trisolini, Luca Bertero, Michela Salvo, Rossella Indellicato, Silvano Andorno, Jose Manuel Garcia-Manteiga, Ottavio Rena, Renzo Luciano Boldorini
AIMS: High-mobility group box-1 (HMGB1) is a chromatin structural protein, ubiquitously expressed in the nuclei of mammalian cells. When transported extracellularly, it acts as tumor suppressor and oncogenic protein. In malignant pleural mesothelioma (MPM), high serum levels of High-mobility group box-1 (HMGB1) have been related to a poor prognosis. Conversely the significance of HMGB1 expression in malignant pleural mesothelioma (MPM) tissues is still unclear. METHODS AND RESULTS: Biopsy samples from 170 patients with MPM were assessed by immunohistochemistry and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) to evaluate HMGB1 protein and gene expression...
January 21, 2018: Histopathology
https://www.readbyqxmd.com/read/29355947/corticotropin-releasing-hormone-binding-protein-is-upregulated-by-brain-derived-neurotrophic-factor-and-is-secreted-in-an-activity-dependent-manner-in-rat-cerebral-cortical-neurons
#3
Naoki Adachi, Shingo Suzuki, Hidetada Matsuoka, Satoko Fushimi, Junichiro Ono, Ken-Ichi Ohta, Yohei Hirai, Takanori Miki, Hisatsugu Koshimizu
A recent study revealed that corticotropin-releasing hormone (CRH) in the cerebral cortex (CTX) plays a regulatory role in emotional behaviors in rodents. Given the functional interaction between brain-derived neurotrophic factor (BDNF) and the CRH-signaling pathway in the hypothalamic-pituitary-adrenal (HPA) axis, we hypothesized that BDNF may regulate gene expression of CRH and its related molecules in the CTX. Findings of real-time quantitative PCR (RT-qPCR) indicated that stimulation of cultured rat cortical neurons with BDNF led to marked elevations in the mRNA levels of CRH and CRH-binding protein (CRH-BP)...
January 22, 2018: Journal of Neurochemistry
https://www.readbyqxmd.com/read/29355533/modulatory-effect-of-glutamate-glur2-receptor-on-the-caudal-neurosecretory-dahlgren-cells-of-the-olive-flounder-paralichthys-olivaceus
#4
Zhaohui Lan, Jinling Xu, Youji Wang, Weiqun Lu
A neuromodulatory role for glutamate has been reported for magnocellular neuroendocrine cells in mammalian hypothalamus. We examined the potential role of glutamate as a local intercellular messenger in the neuroendocrine Dahlgren cell population of the caudal neurosecretory system (CNSS) in the euryhaline flounder Paralichthys olivaceus. In pharmacological experiments in vitro, glutamate (Glu) caused an increase in electrical activity of Dahlgren cells, recruitment of previously silent cells, together with a greater proportion of cells showing phasic (irregular) activity...
January 17, 2018: General and Comparative Endocrinology
https://www.readbyqxmd.com/read/29354959/-effect-of-acupuncture-on-macrophage-polarization-of-white-adipose-tissue-in-obese-mice-induced-by-high-fat-diet
#5
Siyi Zhang, Xia Hu, Hongtu Tang
OBJECTIVE: To observe the effect of acupuncture at acupoints and non-acupoints on macrophage polarization of white adipose tissue in obese mice induced by high-fat diet(HFD). METHODS: Fifty male C57BL/6 mice were randomly divided into a control group (n=10), and other mice were fed with HFD for 16 weeks to establish model. Thirty model mice, which were successfully established and continuously fed with HFD for 8 weeks, were randomized into a model group, a non-acupoint group and an acupuncture group (n=10/group)...
November 12, 2017: Zhongguo Zhen Jiu, Chinese Acupuncture & Moxibustion
https://www.readbyqxmd.com/read/29353007/nitric-oxide-suppresses-aluminum-induced-programmed-cell-death-in-peanut-arachis-hypoganea-l-root-tips-by-improving-mitochondrial-physiological-properties
#6
Huyi He, Wenjing Huang, Thet Lwin Oo, Minghua Gu, Jie Zhan, Aiqin Wang, Long-Fei He
Aluminum (Al) stress alters nitric oxide (NO) and induces programmed cell death (PCD) in plants. Recent study has shown that NO inhibits Al-induced PCD. However, the mechanism of NO inhibiting Al-induced PCD has not been revealed yet. Here, we investigated the behavior of mitochondria during Al-induced PCD suppressed by NO in peanut. Seedlings of peanut was grown hydroponically in a controllable growth room. The mitochondrial physiological parameters were determined spectrophotometrically. The expression of AhANT and AhHsp70 was determined by quantitative RT-PCR...
January 15, 2018: Nitric Oxide: Biology and Chemistry
https://www.readbyqxmd.com/read/29351801/high-expression-of-tmem40-is-associated-with-the-malignant-behavior-and-tumorigenesis-in-bladder-cancer
#7
Zhen-Fei Zhang, Han-Rong Zhang, Qing-Yan Zhang, Shu-Yu Lai, Yu-Zhen Feng, Yi Zhou, Si-Rong Zheng, Rong Shi, Jue-Yu Zhou
BACKGROUND: Bladder cancer (BCa) is one of the most common cancers in the urinary system among the world. Previous studies suggested that TMEM40 expression level was significantly associated with clinicopathological parameters including histological grade, clinical stage and pT status of bladder cancer. However, the molecular mechanism of TMEM40 in BCa remains poorly understood. METHODS: Real-time quantitative RT-PCR (qRT-PCR) and western blot (WB) were used to examine the expression levels of TMEM40 in BCa tissues, paired non-cancer tissues and cell lines...
January 19, 2018: Journal of Translational Medicine
https://www.readbyqxmd.com/read/29350446/differential-gene-expression-in-trigeminal-ganglia-of-male-and-female-rats-following-chronic-constriction-of-the-infraorbital-nerve
#8
O A Korczeniewska, S Husain, J Khan, E Eliav, P Soteropoulos, R Benoliel
BACKGROUND: The mechanisms underlying sex-based differences in pain and analgesia are poorly understood. In this study, we investigated gene expression changes in trigeminal ganglia (TG) of male and female rats exposed to infraorbital nerve chronic constriction injury (IoN-CCI). METHODS: Somatosensory assessments were performed prior to IoN-CCI and at selected time points postsurgery. Selected gene expression changes were examined with real-time quantitative polymerase chain reaction (RT-PCR) in ipsilateral TG at 21 days postsurgery...
January 19, 2018: European Journal of Pain: EJP
https://www.readbyqxmd.com/read/29350427/role-of-microglia-in-the-spinal-cord-in-colon-to-bladder-neural-crosstalk-in-a-rat-model-of-colitis
#9
Tsuyoshi Majima, Yasuhito Funahashi, Naoki Kawamorita, Shun Takai, Yoshihisa Matsukawa, Tokunori Yamamoto, Naoki Yoshimura, Momokazu Gotoh
AIMS: We investigated whether spinal cord microglia are involved in colon-to-bladder neural crosstalk in a rat model of colitis. METHODS: Adult female SD rats were divided into A) control, B) colitis, and C) colitis + minocycline groups. Experimental colitis was induced by administering 50% trinitrobenzene sulfonic acid into the distal colon in the colitis group and the minocycline group. Minocycline, a microglial inhibitor, was continuously infused into the intrathecal space in the minocycline group...
January 19, 2018: Neurourology and Urodynamics
https://www.readbyqxmd.com/read/29349092/cathepsin-b-plays-a-key-role-in-optimal-production-of-the-influenza-a-virus
#10
Macon D Coleman, Soon-Duck Ha, S M Mansour Haeryfar, Stephen Dominic Barr, Sung Ouk Kim
Background: Influenza A virus (IAV) is the etiologic agent of the febrile respiratory illness, commonly referred to as 'flu'. The lysosomal protease cathepsin B (CTSB) has shown to be involved in the lifecycle of various viruses. Here, we examined the role of CTSB in the IAV lifecycle. Methods: CTSB-deficient (CTSB-/-) macrophages and the human lung epithelial cell line A549 cells treated with CA-074Me were infected with the A/Puerto Rico/8/34 strain of IAV (IAV-PR8)...
2018: Journal of Virology & Antiviral Research
https://www.readbyqxmd.com/read/29348885/drug-resistance-in-doxorubicin-resistant-fl5-12-hematopoietic-cells-elevated-mdr1-drug-efflux-and-side-population-positive-and-decreased-bcl2-family-member-expression
#11
Linda S Steelman, Steve L Abrams, Peter Ruvolo, Vivian Ruvolo, Lucio Cocco, Stefano Ratti, Alberto M Martelli, Luca M Neri, Saverio Candido, Massimo Libra, James A McCubrey
Chemotherapeutic drug treatment can result in the emergence of drug-resistant cells. By culturing an interleukin-3 (IL-3)-dependent cell line, FL5.12 cells in the presence of the chemotherapeutic drug doxorubicin, we isolated FL/Doxo cells which are multi-drug resistant. Increased levels of drug efflux were detected in FL/Doxo cells which could be inhibited by the MDR1 inhibitor verapamil but not by the MRP1 inhibitor MK571. The effects of TP53 and MEK1 were examined by infection of FL/Doxo cells with retroviruses encoding either a dominant negative TP-53 gene (FL/Doxo+ TP53 (DN) or a constitutively-activated MEK-1 gene (FL/Doxo + MEK1 (CA)...
December 22, 2017: Oncotarget
https://www.readbyqxmd.com/read/29346643/genetic-characterization-of-a-vang-type-vancomycin-resistant-enterococcus-faecium-clinical-isolate
#12
Mohamed Sassi, François Guérin, Léonie Lesec, Christophe Isnard, Marguerite Fines-Guyon, Vincent Cattoir, Jean-Christophe Giard
Objectives: To characterize, phenotypically and genotypically, the first Enterococcus faecium clinical isolate harbouring a vanG operon. Methods: The antibiotic resistance profile of E. faecium 16-346 was determined and its whole genome sequenced using PacBio technology. Attempts to transfer vancomycin resistance by filter mating were performed and the inducibility of expression of the vanG operon was studied by reverse-transcription quantitative PCR (RT-qPCR) in the presence or absence of subinhibitory concentrations of vancomycin...
January 16, 2018: Journal of Antimicrobial Chemotherapy
https://www.readbyqxmd.com/read/29345197/neonatal-hyperoxia-depletes-pulmonary-vein-cardiomyocytes-in-adult-mice-via-mitochondrial-oxidation
#13
Min Yee, Ethan David Cohen, William Domm, George A Porter, Andrew N McDavid, Michael A O'Reilly
Supplemental oxygen given to preterm infants has been associated with permanently altering postnatal lung development. Now that these individuals are reaching adulthood, there is growing concern that early-life oxygen exposure may also promote cardiovascular disease through poorly understood mechanisms. We previously reported that adult mice exposed to 100% oxygen between postnatal days 0-4 develop pulmonary hypertension defined pathologically by capillary rarefaction, dilation of arterioles and veins, cardiac failure, and a reduced lifespan...
January 18, 2018: American Journal of Physiology. Lung Cellular and Molecular Physiology
https://www.readbyqxmd.com/read/29344870/cell-specific-rna-quantification-in-human-sn-da-neurons-from-heterogeneous-post-mortem-midbrain-samples-by-uv-laser-microdissection-and-rt-qpcr
#14
Johanna Duda, Michael Fauler, Jan Gründemann, Birgit Liss
Cell specificity of gene expression analysis is from particular relevance when the abundance of target cells is not homogeneous in the compared tissue samples, like it is the case, e.g., when comparing brain tissues from controls and in neurodegenerative disease states. While single-cell gene expression profiling is already a methodological challenge per se, it becomes even more prone to artifacts when analyzing individual cells from human post-mortem samples. Not only because human samples can never be matched as precisely as those from animal models, but also, because the RNA-quality that can be obtained from human samples usually displays a high range of variability...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29344858/laser-capture-microdissection-as-a-tool-to-study-the-mucosal-immune-response-in-celiac-disease
#15
Giuseppe Iacomino, Vera Rotondi Aufiero, Pasquale Marena, Antonella Venezia, Riccardo Troncone, Salvatore Auricchio, Giuseppe Mazzarella
Laser capture microdissection (LCM) is a powerful tool for selection and isolation of single cells or compartments from complex primary tissues to perform molecular analyses. Celiac disease is a genetic autoimmune disorder where the ingestion of gluten leads to damage in the small intestine. Increased intraepithelial lymphocytes and the presence of the lamina propria inflammatory infiltrate of the duodenal mucosa is a common part of the disease. These cells promote inflammatory processes through the release of cytokines...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29344852/laser-capture-microdissection-insights-into-methods-and-applications
#16
Meera Mahalingam
Laser capture microdissection is a non-molecular, minimally disruptive method to obtain cytologically and/or phenotypically defined cells or groups of cells from heterogeneous tissues. Its advantages include efficient rapid and precise procurement of cells. The potential disadvantages include time consuming, expensive, and limited by the need for a pathologist for recognition of distinct subpopulations within a specified sample. Overall it is versatile allowing the preparation of homogenous isolates of specific subpopulations of cells from which DNA/RNA or protein can be extracted for RT-PCR, quantitative PCR, next-generation sequencing, immunoblot blot analyses, and mass spectrometry...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29344674/long-non-coding-rna-h1-promotes-cell-proliferation-and-invasion-by-acting-as-a-cerna-of-mir%C3%A2-138-and-releasing-ezh2-in-oral-squamous-cell-carcinoma
#17
Yonglong Hong, Haitao He, Wen Sui, Jingge Zhang, Shenfu Zhang, Dajiang Yang
Long non-coding RNAs (lncRNAs) have been shown to play pivotal roles in various types of human cancer, including oral squamous cell carcinoma (OSCC). However, the potential mechanisms of action of lncRNAs in OSCC remain to be fully elucidated. The aim of the present study was to further explore the potential mechanisms of action of lncRNAs in OSCC. We first analyzed Gene Expression Omnibus (GEO) datasets to investigate aberrantly expressed lncRNAs which may be involved in the development of OSCC. Reverse transcription‑quantitative PCR (RT‑qPCR) was performed to analyze the expression levels of lncRNA H19...
January 16, 2018: International Journal of Oncology
https://www.readbyqxmd.com/read/29343767/blockade-of-trpv1-inhibits-methamphetamine-induced-rewarding-effects
#18
Yu-Hua Tian, Shi-Xun Ma, Kwang-Wook Lee, Sunmee Wee, George F Koob, Seok-Yong Lee, Choon-Gon Jang
Methamphetamine (MAP) is the most widely used psychostimulant in the world, but the exact mechanisms underlying MAP addiction are not yet fully understood. Recent studies have identified the distribution of TRPV1 in several brain regions that are related to drug addiction, including nucleus accumbens (NAc) and dorsal striatum (DSt). In the present study, we performed conditioned place preference (CPP) and self-administration tests to examine the effects of capsazepine (CPZ) and SB366791 (SB) on MAP reward. We found that both CPZ and SB significantly inhibited MAP-induced CPP and self-administration; in contrast, TRPV1 knock-out (KO) mice did not develop MAP-induced CPP...
January 17, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29343665/molecular-characterization-of-gliadins-of-chinese-spring-wheat-in-relation-to-celiac-disease-elicitors
#19
Kanako Kawaura, Mayuko Miura, Yoko Kamei, Tatsuya M Ikeda, Yasunari Ogihara
The wheat seed storage proteins gliadin and glutenin are encoded by multigenes. Gliadins are further classified into α-, γ-, δ- and ω-gliadins. Genes encoding α-gliadins belong to a large multigene family, whose members are located on the homoeologous group 6 chromosomes at the Gli-2 loci. Genes encoding other gliadins are located on the homoeologous group 1 chromosomes at the Gli-1 loci. Two-dimensional polyacrylamide gel electrophoresis (2-DE) was used to characterize and profile the gliadins. The gliadins in aneuploid Chinese Spring wheat lines were then compared in this study...
January 17, 2018: Genes & Genetic Systems
https://www.readbyqxmd.com/read/29343453/polygonatum-sibiricum-polysaccharides-play-anti-cancer-effect-through-tlr4-mapk-nf-%C3%AE%C2%BAb-signaling-pathways
#20
Tingting Long, Zijing Liu, Jingchuan Shang, Xing Zhou, Shuang Yu, Hui Tian, Yixi Bao
OBJECTIVE: To investigate the anti-cancer effect of Polygonatum sibiricum polysaccharides (PSP) and the underlying mechanism. METHODS: Tumor-bearing mice were randomly divided into normal saline (NS) group, adriamycin (ADM) group, PSP group and lipopolysaccharide (LPS) group. RAW264.7 cells were pre-treated with or without TLR4 inhibitor or MyD88 inhibitor. Quantitative RT-PCR and Western blot were performed to detect the mRNA and protein expressions, respectively...
January 14, 2018: International Journal of Biological Macromolecules
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