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Quantitative RT-PCR

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https://www.readbyqxmd.com/read/28102286/chelerythrine-down-regulates-expression-of-vegfa-bcl2-and-kras-by-arresting-g-quadruplex-structures-at-their-promoter-regions
#1
Jagannath Jana, Soma Mondal, Payel Bhattacharjee, Pallabi Sengupta, Tanaya Roychowdhury, Pranay Saha, Pallob Kundu, Subhrangsu Chatterjee
A putative anticancer plant alkaloid, Chelerythrine binds to G-quadruplexes at promoters of VEGFA, BCL2 and KRAS genes and down regulates their expression. The association of Chelerythrine to G-quadruplex at the promoters of these oncogenes were monitored using UV absorption spectroscopy, fluorescence anisotropy, circular dichroism spectroscopy, CD melting, isothermal titration calorimetry, molecular dynamics simulation and quantitative RT-PCR technique. The pronounced hypochromism accompanied by red shifts in UV absorption spectroscopy in conjunction with ethidium bromide displacement assay indicates end stacking mode of interaction of Chelerythrine with the corresponding G-quadruplex structures...
January 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28102059/v%C3%AE-1%C3%A2-%C2%BA-%C3%AE-%C3%AE-t-cells-are-correlated-with-increasing-expression-of-eosinophil-cationic-protein-and-metalloproteinase-7-in-chronic-rhinosinusitis-with-nasal-polyps-inducing-the-formation-of-edema
#2
Luo Ying Yang, Xia Li, Wen Ting Li, Jian Cong Huang, Zhi Yuan Wang, Zi Zhen Huang, Li Hong Chang, Ge Hua Zhang
PURPOSE: We have found that expression of γδT cells is increased in pathological mucosa of chronic rhinosinusitis with nasal polyps (CRSwNP) compared with normal nasal mucosa. This increase is correlated with the infiltration of eosinophils in CRSwNP. Here, we investigated the expression of γδT cells, inflammation and tissue remodeling factors as well as their probable relationships in different types of chronic rhinosinusitis (CRS) in China. METHODS: A total of 76 surgical tissue samples that included 43 CRSwNP samples (15 eosinophilic and 28 non-eosinophilic), 17 CRS samples without nasal polyps (CRSsNP), and 16 controls were obtained...
March 2017: Allergy, Asthma & Immunology Research
https://www.readbyqxmd.com/read/28101577/viscumins-functionally-modulate-cell-motility-associated-gene-expression
#3
Sonja Schötterl, Miriam Hübner, Angela Armento, Vivien Veninga, Naita Maren Wirsik, Simon Bernatz, Hans Lentzen, Michel Mittelbronn, Ulrike Naumann
In Europe extracts from Viscum album L., the European white-berry mistletoe, are widely used as a complementary cancer therapy. Viscumins (mistletoe lectins, ML) have been scrutinized as important active components of mistletoe and exhibit a variety of anticancer effects such as stimulation of the immune system, induction of cytotoxicity, reduction of tumor cell motility as well as changes in the expression of genes associated with cancer development and progression. By microarray expression analysis, quantitative RT-PCR and RT-PCR based validation of microarray data we demonstrate for the Viscum album extract Iscador Qu and for the lectins Aviscumine and ML-1 that in glioma cells these drugs differentially modulate the expression of genes involved in the regulation of cell migration and invasion, including processes modulating cell architecture and cell adhesion...
February 2017: International Journal of Oncology
https://www.readbyqxmd.com/read/28101202/cadherin-11-expression-is-upregulated-in-invasive-human-breast-cancer
#4
Kamil Pohlodek, Yen Y Tan, Christian F Singer, Daphne Gschwantler-Kaulich
Loss of expression of cadherin-11 protein is correlated with a loss of epithelial phenotype and a gain in tumor cell proliferation and invasion. It has been hypothesized that cadherin-11 may be a molecular marker for a more aggressive subtype of breast cancer. The present study examined the expression of the mesenchymal gene/protein cadherin-11 in malignant, benign and healthy breast cancer samples. A paraffin-embedded tissue microarray of both malignant and benign/healthy breast tumor was used. Clinicopathological parameters, including age, grading, tumor size, hormone receptors and HER2 receptors status were obtained from patient medical records...
December 2016: Oncology Letters
https://www.readbyqxmd.com/read/28099495/intestinal-bacteria-composition-and-translocation-of-bacteria-in-inflammatory-bowel-disease
#5
Spyros Vrakas, Konstantinos C Mountzouris, George Michalopoulos, George Karamanolis, George Papatheodoridis, Charalampos Tzathas, Maria Gazouli
BACKGROUND: Live commensal intestinal bacteria are present in the peripheral blood where they can induce inflammation. OBJECTIVE: To evaluate the intestinal bacteria composition and translocation of bacteria in IBD. METHODS: Both blood and tissue biopsy samples were collected from adult patients with active/inactive Crohn's disease (CD), active/inactive ulcerative colitis (UC) and healthy individuals. Most of the patients were newly diagnosed and none of them received antibiotics...
2017: PloS One
https://www.readbyqxmd.com/read/28097816/effect-of-temperature-and-colonization-of-legionella-pneumophila-and-vermamoeba-vermiformis-on-bacterial-community-composition-of-copper-drinking-water-biofilms
#6
Helen Y Buse, Pan Ji, Vicente Gomez-Alvarez, Amy Pruden, Marc A Edwards, Nicholas J Ashbolt
It is unclear how the water-based pathogen, Legionella pneumophila (Lp), and associated free-living amoeba (FLA) hosts change or are changed by the microbial composition of drinking water (DW) biofilm communities. Thus, this study characterized the bacterial community structure over a 7-month period within mature (> 600-day-old) copper DW biofilms in reactors simulating premise plumbing and assessed the impact of temperature and introduction of Lp and its FLA host, Vermamoeba vermiformis (Vv), co-cultures (LpVv)...
January 18, 2017: Microbial Biotechnology
https://www.readbyqxmd.com/read/28096318/bone-marrow-mesenchymal-stem-cells-inhibit-dendritic-cells-differentiation-and-maturation-by-microrna-23b
#7
Jingguo Wu, Chengbo Ji, Qingwei Jia, Feifei Cao, Yong Li, Xishan Zhang, Lanping Wang
BACKGROUND AND AIMS: Research on regulation and its mechanism of bone marrow mesenchymal stem cells (BMSCs) on dendritic cells (DCs) which is the initiating factor in immune response has applicable clinical value. Although BMSCs have a significant regulatory effect on the maturation of DCs, its molecular mechanism is still unclear. METHODS: Isolation and culture of BMSCs and DCs, they were co-cultured in different proportions. Flow cytometry was used to detect the expression of DCs markers (CD83, CD11c)...
January 17, 2017: Bioscience Reports
https://www.readbyqxmd.com/read/28094536/the-novel-induction-of-retinal-ganglion-cell-apoptosis-in-porcine-organ-culture-by-nmda-an-opportunity-for-the-replacement-of-animals-in-experiments
#8
Sandra Kuehn, Jose Hurst, Adelina Jashari, Kathrin Ahrens, Teresa Tsai, Ilan M Wunderlich, H Burkhard Dick, Stephanie C Joachim, Sven Schnichels
Some of the advantages of retina organ culture models include their efficient and easy handling and the ability to standardise relevant parameters. Additionally, when porcine eyes are obtained from the food industry, no animals are killed solely for research purposes. To induce retinal degeneration, a commonly used toxic substance, N-methyl-D-aspartate (NMDA), was applied to the cultures. To this end, organotypic cultures of porcine retinas were cultured and treated with different doses of NMDA (0 [control], 50, 100 and 200μM) on day 2 for 48 hours...
December 2016: Alternatives to Laboratory Animals: ATLA
https://www.readbyqxmd.com/read/28093607/osmopriming-induced-salt-tolerance-during-seed-germination-of-alfalfa-most-likely-mediates-through-h2o2-signaling-and-upregulation-of-heme-oxygenase
#9
Rayhaneh Amooaghaie, Fatemeh Tabatabaie
The present study showed that osmopriming or pretreatment with low H2O2 doses (2 mM) for 6 h alleviated salt-reduced seed germination. The NADPH oxidase activity was the main source, and superoxide dismutase (SOD) activity might be a secondary source of H2O2 generation during osmopriming or H2O2 pretreatment. Hematin pretreatment similar to osmopriming improved salt-reduced seed germination that was coincident with the enhancement of heme oxygenase (HO) activity. The semi-quantitative RT-PCR confirmed that osmopriming or H2O2 pretreatment was able to upregulate heme oxygenase HO-1 transcription, while the application of N,N-dimethyl thiourea (DMTU as trap of endogenous H2O2) and diphenyleneiodonium (DPI as inhibitor of NADPHox) not only blocked the upregulation of HO but also reversed the osmopriming-induced salt attenuation...
January 16, 2017: Protoplasma
https://www.readbyqxmd.com/read/28093213/characterization-of-the-efficiency-and-uncertainty-of-skimmed-milk-flocculation-for-the-simultaneous-concentration-and-quantification-of-water-borne-viruses-bacteria-and-protozoa
#10
Eloy Gonzales-Gustavson, Yexenia Cardenas-Youngs, Miquel Calvo, Marcelle Figueira Marques da Silva, Ayalkibet Hundesa, Inmaculada Amorós, Yolanda Moreno, Laura Moreno-Mesonero, Rosa Rosell, Lliliane Ganges, Rosa Araujo, Rosina Girones
In this study, the use of skimmed milk flocculation (SMF) to simultaneously concentrate viruses, bacteria and protozoa was evaluated. We selected strains of faecal indicator bacteria and pathogens, such as Escherichia coli and Helicobacter pylori. The viruses selected were adenovirus (HAdV 35), rotavirus (RoV SA-11), the bacteriophage MS2 and bovine viral diarrhoea virus (BVDV). The protozoa tested were Acanthamoeba, Giardia and Cryptosporidium. The mean recoveries with q(RT)PCR were 66% (HAdV 35), 24% (MS2), 28% (RoV SA-11), 15% (BVDV), 60% (E...
January 13, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28091410/molecular-mechanisms-of-increased-heart-rate-in-shenxianshengmai-treated-bradycardia-rabbits
#11
Zhou-Ying Liu, Jian Huang, Na-Na Liu, Min Zheng, Tao Zhao, Bu-Chang Zhao, Yi-Min Wang, Jie-Lin Pu
BACKGROUND: The molecular mechanisms of Shenxianshengmai (SXSM), a traditional Chinese medicine, on bradycardia have been incompletely understood. The study tried to investigate the gene expression profile and proteomics of bradycardia rabbits' hearts after SXSM treatment. METHODS: Twenty-four adult rabbits were randomly assigned in four groups: sham, model, model plus SXSM treatment, and sham plus SXSM treatment groups. Heart rate was recorded in all rabbits. Then, total RNA of atria and proteins of ventricle were isolated and quantified, respectively...
2017: Chinese Medical Journal
https://www.readbyqxmd.com/read/28089820/deregulated-expression-of-vhl-mrna-variants-in-papillary-thyroid-cancer
#12
Enke Baldini, Chiara Tuccilli, Yannick Arlot-Bonnemains, Frank Chesnel, Salvatore Sorrenti, Corrado De Vito, Antonio Catania, Eleonora D'Armiento, Alessandro Antonelli, Poupak Fallahi, Sara Watutantrige-Fernando, Francesco Tartaglia, Susi Barollo, Caterina Mian, Marco Bononi, Stefano Arceri, Domenico Mascagni, Massimo Vergine, Daniele Pironi, Massimo Monti, Angelo Filippini, Salvatore Ulisse
Recent findings demonstrated that a subset of papillary thyroid cancers (PTCs) is characterized by reduced expression of the von Hippel-Lindau (VHL) tumor suppressor gene, and that lowest levels associated with more aggressive PTCs. In the present study, the levels of the two VHL mRNA splicing variants, VHL-213 (V1) and VHL-172 (V2), were measured in a series of 96 PTC and corresponding normal matched tissues by means of quantitative RT-PCR. Variations in the mRNA levels were correlated with patients' clinicopathological parameters and disease-free interval (DFI)...
January 12, 2017: Molecular and Cellular Endocrinology
https://www.readbyqxmd.com/read/28089781/genotoxic-effect-and-rat-hepatocyte-death-occurred-after-oxidative-stress-induction-and-antioxidant-gene-downregulation-caused-by-long-term-fluoride-exposure
#13
F D Campos-Pereira, L Lopes-Aguiar, F L Renosto, G A S Nogueira, E F D Costa, R Barbieri Pulz, E C M Silva-Zacarin, C A Oliveira, A A Pigoso, G D C Severi-Aguiar
Studies focusing on possible genotoxic effects of excess fluoride are contradictory and inconclusive. Currently, studies have reported a probable link to oxidative stress, DNA damage and apoptosis induced by fluoride in rat hepatocytes. We developed an in vivo study administering three doses of fluoride by gavage given to rats for 60 day. Micronucleus test was applied to investigate genotoxic potential of fluoride. The TUNEL method determined DNA fragmentation and apoptosis. Biochemical parameters to investigate mitochondrial swelling and oxidative stress...
January 12, 2017: Chemico-biological Interactions
https://www.readbyqxmd.com/read/28089740/early-remodeling-of-repolarizing-k-currents-in-the-%C3%AE-mhc-403-mouse-model-of-familial-hypertrophic-cardiomyopathy
#14
Rocco Hueneke, Adam Adenwala, Rebecca L Mellor, Jonathan G Seidman, Christine E Seidman, Jeanne M Nerbonne
Familial hypertrophic cardiomyopathy (HCM), linked to mutations in myosin, myosin-binding proteins and other sarcolemmal proteins, is associated with increased risk of life threatening ventricular arrhythmias, and a number of animal models have been developed to facilitate analysis of disease progression and mechanisms. In the experiments here, we use the αMHC(403/+) mouse line in which one αMHC allele harbors a common HCM mutation (in βMHC, Arg403 Gln). Here, we demonstrate marked prolongation of QT intervals in young adult (10-12week) male αMHC(403/+) mice, well in advance of the onset of measurable left ventricular hypertrophy...
January 12, 2017: Journal of Molecular and Cellular Cardiology
https://www.readbyqxmd.com/read/28089386/detection-of-urinary-cell-free-mir-210-as-a-potential-tool-of-liquid-biopsy-for-clear-cell-renal-cell-carcinoma
#15
Guorong Li, An Zhao, Michel Péoch, Michèle Cottier, Nicolas Mottet
BACKGROUND: Recent findings show that cell-free miRNAs are stable in biological fluid. Urine provides an alternative to blood serum or plasma as a potential source of tumor biomarkers. MiR-210 is proven to be overexpressed in patients with a clear cell renal cell carcinoma (ccRCC). The purpose of this pilot study was to examine the urinary cell-free miR-210 as a potential tool of liquid biopsy for ccRCC. METHODS: Overall, 75 patients with a ccRCC and 45 control subjects without a cancer were included in this study...
January 12, 2017: Urologic Oncology
https://www.readbyqxmd.com/read/28088972/-critical-roles-of-matrix-metalloproteinases-secreted-by-leukemic-cells-in-the-pathogenesis-of-central-nervous-system-leukemia
#16
S R Feng, Z X Chen, J N Cen, H J Shen, Y Y Wang, L Yao
Objective: To observe the effects of matrix metalloproteinases (MMP)-2 and MMP-9 secreted by leukemic cells on tight junction proteins ZO-1, claudin-5 and occluding and the permeability of the blood-brain barrier (BBB) and explore the mechanisms of MMP-2 and MMP-9 in leukemic cell infiltration of the central nervous system (CNS). Methods: The mRNA expressions of MMP-2 and MMP-9 in leukemic cell lines SHI-1, HL-60 and U937 were detected by quantitative RT-PCR. The MMP inhibitor GM6001 was used to inhibit the secretion of MMP-2 and MMP-9...
December 14, 2016: Zhonghua Xue Ye Xue za Zhi, Zhonghua Xueyexue Zazhi
https://www.readbyqxmd.com/read/28088556/regulation-of-docetaxel-sensitivity-in-prostate-cancer-cells-by-hsa-mir-125a-3p-via-modulation-of-mta1-signaling
#17
Jian-Zhou Liu, Feng-Yan Yin, Chang-You Yan, Hui Wang, Xiao-Hui Luo
OBJECTIVE: To identify the potential down-stream targets of hsa-miR-125a-3p, a mature form of miR-125a, during the pathogenesis of chemoresistance in prostate cancer (PCa). METHODS: Expression levels of hsa-miR-125a-3p was assessed in chemoresistant PCa tissues and experimentally-established chemoresistant cells using quantitative RT-PCR (qRT-PCR). The effect of hsa-miR-125a-3p knockdown or hsa-miR-125a-3p overexpression on the Dox-induced cell death was evaluated using apoptosis ELISA in chemosensitive PC-3 cells or in chemoresistant PC-3 cells (PC-3R)...
January 11, 2017: Urology
https://www.readbyqxmd.com/read/28088212/ultrasensitive-detection-of-oncogenic-human-papillomavirus-in-oropharyngeal-tissue-swabs
#18
Andre Isaac, Morris Kostiuk, Han Zhang, Cameron Lindsay, Fawaz Makki, Daniel A O'Connell, Jeffrey R Harris, David W J Cote, Hadi Seikaly, Vincent L Biron
BACKGROUND: The incidence of oropharyngeal squamous cell carcinoma (OPSCC) caused by oncogenic human papillomavirus (HPV) is rising worldwide. HPV-OPSCC is commonly diagnosed by RT-qPCR of HPV E6 and E7 oncoproteins or by p16 immunohistochemistry (IHC). Droplet digital PCR (ddPCR) has been recently reported as an ultra-sensitive and highly precise method of nucleic acid quantification for biomarker analysis. To validate the use of a minimally invasive assay for detection of oncogenic HPV based on oropharyngeal swabs using ddPCR...
January 14, 2017: Journal of Otolaryngology—Head & Neck Surgery
https://www.readbyqxmd.com/read/28087386/androgens-and-androgen-receptor-signaling-contribute-to-ovarian-development-in-the-chicken-embryo
#19
Ryoma Tanaka, Hiroe Izumi, Asato Kuroiwa
Androgens and androgen receptor (AR) signaling play important roles throughout development. In the chicken, AR signaling is involved in reproduction; however, its specific role is unclear. We show that AR signaling is involved in the normal development of the female embryonic gonads. The AR mRNA level was detected in male and female embryonic gonads by quantitative RT-PCR, and its expression was higher in females than in males at all developmental stages examined. In female embryos, the AR localized to nuclei of cells in the left gonad...
January 10, 2017: Molecular and Cellular Endocrinology
https://www.readbyqxmd.com/read/28086912/wwl70-attenuates-pge2-production-derived-from-2-arachidonoylglycerol-in-microglia-by-abhd6-independent-mechanism
#20
Mikiei Tanaka, Sean Moran, Jie Wen, Kwame Affram, Tinghua Chen, Aviva J Symes, Yumin Zhang
BACKGROUND: α/β-Hydrolase domain 6 (ABHD6) is one of the major enzymes for endocannabinoid 2-arachidonoylglycerol (2-AG) hydrolysis in microglia cells. Our recent studies have shown that a selective ABHD6 inhibitor WWL70 has anti-inflammatory and neuroprotective effects in animal models of traumatic brain injury and multiple sclerosis. However, the role of ABHD6 in the neuroinflammatory response and the mechanisms by which WWL70 suppresses inflammation has not yet been elucidated in reactive microglia...
January 10, 2017: Journal of Neuroinflammation
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