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https://www.readbyqxmd.com/read/28332571/profiling-protein-expression-in-circulating-tumour-cells-using-microfluidic-western-blotting
#1
Elly Sinkala, Elodie Sollier-Christen, Corinne Renier, Elisabet Rosàs-Canyelles, James Che, Kyra Heirich, Todd A Duncombe, Julea Vlassakis, Kevin A Yamauchi, Haiyan Huang, Stefanie S Jeffrey, Amy E Herr
Circulating tumour cells (CTCs) are rare tumour cells found in the circulatory system of certain cancer patients. The clinical and functional significance of CTCs is still under investigation. Protein profiling of CTCs would complement the recent advances in enumeration, transcriptomic and genomic characterization of these rare cells and help define their characteristics. Here we describe a microfluidic western blot for an eight-plex protein panel for individual CTCs derived from estrogen receptor-positive (ER+) breast cancer patients...
March 23, 2017: Nature Communications
https://www.readbyqxmd.com/read/28331561/imatinib-induces-up-regulation-of-nm23-a-metastasis-suppressor-gene-in-human-hepatocarcinoma-hepg2-cell-line
#2
Behta Keshavarz-Pakseresht, Seyed Ataollah Sadat Shandiz, Fahimeh Baghbani-Arani
AIM: The present study investigated the anti-tumor activity of Imatinib mesylate through modulation of NM23 gene expression in human hepatocellular carcinoma (HepG2) cell line. BACKGROUND: Hepatocellular carcinoma (HCC) is considered to be the third leading cause of cancer related death worldwide. Down regulation of NM23, a metastasis suppressor gene, has been associated with several types of malignant cancer. Recently, effects of Imatinib mesylate, a first member of tyrosine kinases inhibitors, were indicated in research and treatment of different malignant tumors...
2017: Gastroenterology and Hepatology From Bed to Bench
https://www.readbyqxmd.com/read/28328930/validation-of-qpcr-reference-genes-in-lymphocytes-from-patients-with-amyotrophic-lateral-sclerosis
#3
Ewa Usarek, Anna Barańczyk-Kuźma, Beata Kaźmierczak, Beata Gajewska, Magdalena Kuźma-Kozakiewicz
Quantitative polymerase chain reaction (qPCR) is the most specific and reliable method for determination of mRNA gene expression. Crucial point for its accurate normalization is the choice of appropriate internal control genes (ICGs). In the present work we determined and compare the expression of eight commonly used ICGs in lymphocytes from 26 patients with amyotrophic lateral sclerosis (ALS) and 30 control subjects. Peripheral blood mononuclear cells (PBMCs) before and after immortalization by EBV transfection (lymphoblast cell lines-LCLs) were used for qPCR analysis...
2017: PloS One
https://www.readbyqxmd.com/read/28328240/insights-on-cryopreserved-sheep-fibroblasts-by-cryomicroscopy-and-gene-expression-analysis
#4
Antonella Idda, Daniela Bebbere, Giuseppina Corona, Laura Masala, Elisa Casula, Alberto Cincotti, Sergio Ledda
Cryopreservation includes a set of techniques aimed at storing biological samples and preserving their biochemical and functional features without any significant alterations. This study set out to investigate the effects induced by cryopreservation on cultured sheepskin fibroblasts (CSSF) through cryomicroscopy and gene expression analysis after subsequent in vitro culture. CSSF cells were cryopreserved in a cryomicroscope (CM) or in a straw programmable freezer (SPF) using a similar thermal profile (cooling rate -5°C/min to -120°C, then -150°C/min to -196°C)...
March 22, 2017: Biopreservation and Biobanking
https://www.readbyqxmd.com/read/28327178/glyceraldehyde-3-phosphate-dehydrogenase-acts-as-an-adhesin-in-erysipelothrix-rhusiopathiae-adhesion-to-porcine-endothelial-cells-and-as-a-receptor-in-recruitment-of-host-fibronectin-and-plasminogen
#5
Weifeng Zhu, Qiang Zhang, Jingtao Li, Yanmin Wei, Chengzhi Cai, Liang Liu, Zhongmin Xu, Meilin Jin
Erysipelothrix rhusiopathiae is the causative agent of animal erysipelas and human erysipeloid. Previous studies suggested glyceraldehyde 3-phosphate dehydrogenase (GAPDH) plays a role in the pathogenesis of E. rhusiopathiae infection. We studied E. rhusiopathiae GAPDH interactions with pig vascular endothelial cells, fibronectin, and plasminogen. Recombinant GAPDH (rGAPDH) was successfully obtained, and it was shown that it plays a role in E. rhusiopathiae adhesion to pig vascular endothelial cells. Moreover, rGAPDH could bind fibronectin and plasminogen in a dose-dependent manner...
March 21, 2017: Veterinary Research
https://www.readbyqxmd.com/read/28323834/selection-and-validation-of-reference-genes-for-qrt-pcr-analysis-during-biological-invasions-the-thermal-adaptability-of-bemisia-tabaci-med
#6
Tian-Mei Dai, Zhi-Chuang Lü, Wan-Xue Liu, Fang-Hao Wan
The Bemisia tabaci Mediterranean (MED) cryptic species has been rapidly invading to most parts of the world owing to its strong ecological adaptability, which is considered as a model insect for stress tolerance studies under rapidly changing environments. Selection of a suitable reference gene for quantitative stress-responsive gene expression analysis based on qRT-PCR is critical for elaborating the molecular mechanisms of thermotolerance. To obtain accurate and reliable normalization data in MED, eight candidate reference genes (β-act, GAPDH, β-tub, EF1-α, GST, 18S, RPL13A and α-tub) were examined under various thermal stresses for varied time periods by using geNorm, NormFinder and BestKeeper algorithms, respectively...
2017: PloS One
https://www.readbyqxmd.com/read/28315300/interaction-of-glyceraldehyde-3-phosphate-dehydrogenase-and-heme-the-relevance-of-its-biological-function
#7
Yi Huang, Pengfei Zhang, Zhen Yang, Peipei Wang, Hailing Li, Zhonghong Gao
GAPDH was speculated to function as a transient trap to reduce the potential toxicity of free heme by a specific and reversible binding with heme. Up to now, there has been lack of studies focused on this effect. In this paper, the efficiency of GAPDH-heme complex on catalyzing protein carbonylation and nitration, the cross-linking of heme to protein formation, and cytotoxicity of GAPDH-heme were studied. It was found that the binding of GAPDH could inhibit H2O2-mediated degradation of heme. Peroxidase activity of GAPDH-heme complex was higher than that of free heme, but significantly lower than that of HSA-heme...
April 1, 2017: Archives of Biochemistry and Biophysics
https://www.readbyqxmd.com/read/28301553/multiplex-pcr-as-a-tool-for-the-diagnosis-of-leishmania-spp-kdna-and-the-gapdh-housekeeping-gene-of-mammal-hosts
#8
Renata de Cássia-Pires, Myllena de Fátima Alheiros Dias de Melo, Raquel da Hora Barbosa, André Luiz Rodrigues Roque
BACKGROUND: The PCR assays usually employed for Leishmania diagnosis does not simultaneously detect a constitutive gene that would certify the viability of the DNA sample. We present a multiplex PCR approach for the simultaneous diagnosis of the Leishmania sp. kDNA fragment and a catalytic domain segment of a conserved region of the mammalian gapdh gene. METHODOLOGY: The proposed multiplex protocol was designed through in silico PCR. The annealing temperature, concentration of primer pairs, number of cycles, distinct polymerase enzymes and premix kit were defined to achieve an optimal reaction...
2017: PloS One
https://www.readbyqxmd.com/read/28301521/lysosomal-adaptation-how-cells-respond-to-lysosomotropic-compounds
#9
Shuyan Lu, Tae Sung, Nianwei Lin, Robert T Abraham, Bart A Jessen
Lysosomes are acidic organelles essential for degradation and cellular homoeostasis and recently lysosomes have been shown as signaling hub to respond to the intra and extracellular changes (e.g. amino acid availability). Compounds including pharmaceutical drugs that are basic and lipophilic will become sequestered inside lysosomes (lysosomotropic). How cells respond to the lysosomal stress associated with lysosomotropism is not well characterized. Our goal is to assess the lysosomal changes and identify the signaling pathways that involve in the lysosomal changes...
2017: PloS One
https://www.readbyqxmd.com/read/28300076/evaluation-of-reference-genes-at-different-developmental-stages-for-quantitative-real-time-pcr-in-aedes-aegypti
#10
Najat Dzaki, Karima N Ramli, Azali Azlan, Intan H Ishak, Ghows Azzam
The mosquito Aedes aegypti (Ae. aegypti) is the most notorious vector of illness-causing viruses such as Dengue, Chikugunya, and Zika. Although numerous genetic expression studies utilizing quantitative real-time PCR (qPCR) have been conducted with regards to Ae. aegypti, a panel of genes to be used suitably as references for the purpose of expression-level normalization within this epidemiologically important insect is presently lacking. Here, the usability of seven widely-utilized reference genes i.e. actin (ACT), eukaryotic elongation factor 1 alpha (eEF1α), alpha tubulin (α-tubulin), ribosomal proteins L8, L32 and S17 (RPL8, RPL32 and RPS17), and glyceraldeyde 3-phosphate dehydrogenase (GAPDH) were investigated...
March 16, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28299307/identification-of-appropriate-housekeeping-genes-for-gene-expression-analysis-in-long-term-hypoxia-treated-kidney-cells
#11
Shiva Moein, Shaghayegh Haghjooy Javanmard, Maryam Abedi, Mohammad Hosein Izadpanahi, Yousof Gheisari
BACKGROUND: Selection of stably expressing housekeeping genes (HKGs) is a crucial step in gene expression analysis. However, there are no universal HKGs for all experiments, and they should be determined by each biologic condition. The aim of this study was to detect appropriate HKGs for kidney cells cultured in long-term hypoxia. MATERIALS AND METHODS: Based on a screening step using a microarray data available from gene expression omnibus database, a set of candidate HKGs were chosen to be assessed in human kidney cells cultured in hypoxic or normoxic conditions for about 2 weeks in a time course manner...
2017: Advanced Biomedical Research
https://www.readbyqxmd.com/read/28298621/silencing-nc886-a-non-coding-rna-induces-apoptosis-of-human-endometrial-cancer-cells-1a-in-vitro
#12
Zhuoying Hu, Hongyu Zhang, Liangdan Tang, Meng Lou, Yanqing Geng
BACKGROUND The role that nc886, a non-coding microRNA, plays in human endometrial cancer is unknown. The present study aimed to describe the functional role of nc886 in human endometrial cancer-1A (HEC-1A) cell line, which may provide another target for human endometrial cancer treatment. MATERIAL AND METHODS The expression levels of nv886 in normal human endometrial tissue and the early phase and late phase of human endometrial cancer tissues were determined and compared by fluorescence in situ hybridization (FISH)...
March 16, 2017: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
https://www.readbyqxmd.com/read/28298336/moonlighting-glycolytic-protein-glyceraldehyde-3-phosphate-dehydrogenase-gapdh-an-evolutionarily-conserved-plasminogen-receptor-on-mammalian-cells
#13
Anoop Singh Chauhan, Manoj Kumar, Surbhi Chaudhary, Anil Patidar, Asmita Dhiman, Navdeep Sheokand, Himanshu Malhotra, Chaaya Iyengar Raje, Manoj Raje
Prokaryotic pathogens establish infection in mammals by capturing the proteolytic enzyme plasminogen (Plg) onto their surface to digest host extracellular matrix (ECM). One of the bacterial surface Plg receptors is the multifunctional glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). In a defensive response, the host mounts an inflammatory response, which involves infiltration of leukocytes to sites of inflammation. This requires macrophage exit from the blood and migration across basement membranes, a phenomenon dependent on proteolytic remodeling of the ECM utilizing Plg...
March 15, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/28294410/reference-gene-selection-for-reverse-transcription-quantitative-polymerase-chain-reaction-in-chicken-hypothalamus-under-different-feeding-status
#14
Á Simon, A Jávor, P Bai, J Oláh, L Czeglédi
This study was designed to investigate the stability of 10 candidate reference genes, namely ACTB, B2M, GAPDH, HMBS, LBR, POLR2B, RN18S, RPS17, TBP, and YWHAZ for the normalization of gene expression data obtained by quantitative real-time polymerase chain reaction (qPCR) in studies related to feed intake of chicken. Samples were isolated from hypothalamus under three different nutritional status (ad libitum, fasted for 24 hr, fasted for 24 hr then refed for 2 hr). Five different algorithms were applied for the analysis of reference gene stability: BestKeeper, geNorm, NormFinder, the comparative ΔCt method, and a novel approach using multivariate linear mixed-effects modelling for stable reference gene selection...
March 15, 2017: Journal of Animal Physiology and Animal Nutrition
https://www.readbyqxmd.com/read/28285168/design-and-use-of-mouse-control-dna-for-dna-biomarker-extraction-and-pcr-detection-from-urine-application-for-transrenal-mycobacterium-tuberculosis-dna-detection
#15
Hali Bordelon, Keersten M Ricks, Megan E Pask, Patricia K Russ, Francesca Solinas, Mark L Baglia, Philip A Short, Andrew Nel, Jonathan Blackburn, Keertan Dheda, Carlos Zamudio, Tatiana Cáceres, David W Wright, Frederick R Haselton, April C Pettit
Urine samples are increasingly used for diagnosing infections including Escherichia coli, Ebola virus, and Zika virus. However, extraction and concentration of nucleic acid biomarkers from urine is necessary for many molecular detection strategies such as polymerase chain reaction (PCR). Since urine samples typically have large volumes with dilute biomarker concentrations making them prone to false negatives, another impediment for urine-based diagnostics is the establishment of appropriate controls particularly to rule out false negatives...
March 9, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28284075/a-fluorogenic-probe-for-imaging-protein-s-nitrosylation-in-live-cells
#16
Shiyi Shao, Bo Chen, Juan Cheng, Chengkun Wang, Yanli Zhang, Lingxiao Shao, Yongzhou Hu, Yifeng Han, Feng Han, Xin Li
S-nitrosylation is a posttranslational modification of protein cysteine residues leading to the formation of S-nitrosothiols and its detection is crucial to understanding of redox regulation and NO-based signaling. Prototypical detection methods for S-nitrosylation are always carried out ex situ. However, the reversible nature and the tendency of transnitrosylation highlight the necessity of its probing in intact live biological contexts. Herein we provide a fluorogenic chemical probe for the detection of S-nitrosylation in live endothelial cells...
March 1, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28271485/d-glyceraldehyde-3-phosphate-dehydrogenase-structure-and-function
#17
Michael R White, Elsa D Garcin
Aside from its well-established role in glycolysis, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has been shown to possess many key functions in cells. These functions are regulated by protein oligomerization , biomolecular interactions, post-translational modifications , and variations in subcellular localization . Several GAPDH functions and regulatory mechanisms overlap with one another and converge around its role in intermediary metabolism. Several structural determinants of the protein dictate its function and regulation...
2017: Sub-cellular Biochemistry
https://www.readbyqxmd.com/read/28265765/physiology-phylogeny-early-evolution-and-gapdh
#18
REVIEW
William F Martin, Rüdiger Cerff
The chloroplast and cytosol of plant cells harbor a number of parallel biochemical reactions germane to the Calvin cycle and glycolysis, respectively. These reactions are catalyzed by nuclear encoded, compartment-specific isoenzymes that differ in their physiochemical properties. The chloroplast cytosol isoenzymes of D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) harbor evidence of major events in the history of life: the origin of the first genes, the bacterial-archaeal split, the origin of eukaryotes, the evolution of protein compartmentation during eukaryote evolution, the origin of plastids, and the secondary endosymbiosis among the algae with complex plastids...
March 6, 2017: Protoplasma
https://www.readbyqxmd.com/read/28261247/selection-of-candidate-reference-genes-for-gene-expression-analysis-in-kentucky-bluegrass-poa-pratensis-l-under-abiotic-stress
#19
Kuiju Niu, Yi Shi, Huiling Ma
Kentucky bluegrass (Poa pratensis L.) belong to Gramineae and is widely used in lawns, golf courses, landscapes, and sport fields as a prominent cool-season grass. Gene expression patterns during different stages of plant development can provide clues toward the understanding of its biological functions. The selection and validation of reference genes are the first steps in any real-time quantitative PCR gene expression study. Therefore, suitable reference genes are necessary for obtaining reliable results in real-time quantitative PCR analyses of Kentucky bluegrass...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28258188/lysine-post-translational-modification-of-glyceraldehyde-3-phosphate-dehydrogenase-regulates-hepatic-and-systemic-metabolism
#20
Simon T Bond, Kirsten F Howlett, Greg M Kowalski, Shaun Mason, Timothy Connor, Adrian Cooper, Victor Streltsov, Clinton R Bruce, Ken R Walder, Sean L McGee
Reciprocal regulation of hepatic glycolysis and gluconeogenesis contributes to systemic metabolic homeostasis. Recent evidence from lower order organisms has found that reversible post-translational modification of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), particularly acetylation, contributes to the reciprocal regulation of glycolysis/gluconeogenesis. However, whether this occurs in mammalian hepatocytes in vitro or in vivo is unknown. Several proteomics studies have identified 4 lysine residues in critical regions of mammalian GAPDH that are altered by multiple post-translational modifications...
March 3, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
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