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https://www.readbyqxmd.com/read/28935607/s-glutathionylation-of-glyceraldehyde-3-phosphate-dehydrogenase-induces-formation-of-c150-c154-intrasubunit-disulfide-bond-in-the-active-site-of-the-enzyme
#1
K V Barinova, M V Serebryakova, V I Muronetz, E V Schmalhausen
BACKGROUND: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a glycolytic protein involved in numerous non-glycolytic functions. S-glutathionylated GAPDH was revealed in plant and animal tissues. The role of GAPDH S-glutathionylation is not fully understood. METHODS: Rabbit muscle GAPDH was S-glutathionylated in the presence of H2O2 and reduced glutathione (GSH). The modified protein was assayed by MALDI-MS analysis, differential scanning calorimetry, dynamic light scattering, and ultracentrifugation...
September 18, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28919905/candidate-reference-genes-selection-and-application-for-rt-qpcr-analysis-in-kenaf-with-cytoplasmic-male-sterility-background
#2
Bujin Zhou, Peng Chen, Aziz Khan, Yanhong Zhao, Lihong Chen, Dongmei Liu, Xiaofang Liao, Xiangjun Kong, Ruiyang Zhou
Cytoplasmic male sterility (CMS) is a maternally inherited trait that results in the production of dysfunctional pollen. Based on reliable reference gene-normalized real-time quantitative PCR (RT-qPCR) data, examining gene expression profile can provide valuable information on the molecular mechanism of kenaf CMS. However, studies have not been conducted regarding selection of reference genes for normalizing RT-qPCR data in the CMS and maintainer lines of kenaf crop. Therefore, we studied 10 candidate reference genes (ACT3, ELF1A, G6PD, PEPKR1, TUB, TUA, CYP, GAPDH, H3, and 18S) to assess their expression stability at three stages of pollen development in CMS line 722A and maintainer line 722B of kenaf...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28918937/a-predictive-model-for-selective-targeting-of-the-warburg-effect-through-gapdh-inhibition-with-a-natural-product
#3
Maria V Liberti, Ziwei Dai, Suzanne E Wardell, Joshua A Baccile, Xiaojing Liu, Xia Gao, Robert Baldi, Mahya Mehrmohamadi, Marc O Johnson, Neel S Madhukar, Alexander A Shestov, Iok I Christine Chio, Olivier Elemento, Jeffrey C Rathmell, Frank C Schroeder, Donald P McDonnell, Jason W Locasale
Targeted cancer therapies that use genetics are successful, but principles for selectively targeting tumor metabolism that is also dependent on the environment remain unknown. We now show that differences in rate-controlling enzymes during the Warburg effect (WE), the most prominent hallmark of cancer cell metabolism, can be used to predict a response to targeting glucose metabolism. We establish a natural product, koningic acid (KA), to be a selective inhibitor of GAPDH, an enzyme we characterize to have differential control properties over metabolism during the WE...
September 8, 2017: Cell Metabolism
https://www.readbyqxmd.com/read/28917607/selection-of-reliable-reference-genes-for-gene-expression-studies-in-botrytis-cinerea
#4
He Ren, Xiaoqing Wu, Yuping Lyu, Hongzi Zhou, Xueying Xie, Xinjian Zhang, Hetong Yang
Botrytis cinerea is an important plant pathogen causing grey mold disease in a wide range of plant species. The aim of this study was to identify reliable reference genes that can be used for the analysis of relative gene expression in B. cinerea with quantitative real-time reverse transcription PCR (qRT-PCR). Six commonly used housekeeping genes including actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ubiquitin (UBQ), ubiquitin-conjugating enzyme (UCE), α-tubulin (α-TUB) and β-tubulin (β-TUB) were selected to test their expression stabilities in B...
September 14, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28916476/pharmacologic-ascorbate-induces-neuroblastoma-cell-death-by-hydrogen-peroxide-mediated-dna-damage-and-reduction-in-cancer-cell-glycolysis
#5
Enlong Ma, Ping Chen, Heather M Wilkins, Tao Wang, Russell H Swerdlow, Qi Chen
An ascorbate-mediated production of oxidative stress has been shown to retard tumor growth. Subsequent glycolysis inhibition has been suggested. Here, we further define the mechanisms relevant to this observation. Ascorbate was cytotoxic to human neuroblastoma cells through the production of H2O2, which led to ATP depletion, inhibited GAPDH, and non-apoptotic and non-autophagic cell death. The mechanism of cytotoxicity is different when PARP-dependent DNA repair machinery is active or inhibited. Ascorbate-generated H2O2 damaged DNA, activated PARP, depleted NAD+, and reduced glycolysis flux...
September 12, 2017: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/28914500/comprehensive-proteomics-analysis-of-exosomes-derived-from-human-seminal-plasma
#6
C Yang, W-B Guo, W-S Zhang, J Bian, J-K Yang, Q-Z Zhou, M-K Chen, W Peng, T Qi, C-Y Wang, C-D Liu
Exosomes are membranous nanovesicles of endocytic origin that carry and transfer regulatory bioactive molecules and mediate intercellular communication between cells and tissues. Although seminal exosomes have been identified in human seminal plasma, their exact composition and possible physiologic function remain unknown. The objective of this study was to perform a comprehensive proteomics analysis of exosomes derived from human seminal plasma. Seminal exosomes were isolated and purified from 12 healthy donors using a 30% sucrose cushion-based exosome-isolation protocol, followed by characterization by western blot, transmission electron microscopy, and nanoparticle tracking analysis before performing extensive liquid chromatography tandem mass spectrometry proteomics analysis...
September 2017: Andrology
https://www.readbyqxmd.com/read/28910197/optimal-reference-genes-for-rt-qpcr-normalization-in-the-newborn
#7
P Khanna, K L Johnson, J L Maron
It is difficult to identify reliable reference genes for transcriptomic analyses in biofluids such as saliva. This situation is particularly relevant for the newborn population, where rapid development is associated with dynamic changes in gene expression. Real-time gene expression monitoring holds great promise for elucidating disrupted pathways that result in morbidities unique to this population, such as retinopathy of prematurity, but its impact depends on identifying stable and consistently expressed genes across a wide range of gestational ages...
September 14, 2017: Biotechnic & Histochemistry: Official Publication of the Biological Stain Commission
https://www.readbyqxmd.com/read/28905328/lessons-learned-from-protein-aggregation-toward-technological-and-biomedical-applications
#8
REVIEW
César L Avila, Silvina Chaves, Sergio B Socias, Esteban Vera-Pingitore, Florencia González-Lizárraga, Cecilia Vera, Diego Ploper, Rosana Chehín
The close relationship between protein aggregation and neurodegenerative diseases has been the driving force behind the renewed interest in a field where biophysics, neurobiology and nanotechnology converge in the study of the aggregate state. On one hand, knowledge of the molecular principles that govern the processes of protein aggregation has a direct impact on the design of new drugs for high-incidence pathologies that currently can only be treated palliatively. On the other hand, exploiting the benefits of protein aggregation in the design of new nanomaterials could have a strong impact on biotechnology...
September 13, 2017: Biophysical Reviews
https://www.readbyqxmd.com/read/28905096/comparison-of-suction-blistering-and-tape-stripping-for-analysis-of-epidermal-genes-proteins-and-lipids
#9
Marek Svoboda, Marta Hlobilová, Markéta Marešová, Michaela Sochorová, Andrej Kováčik, Kateřina Vávrová, Iva Dolečková
Analysis of epidermal genes, proteins and lipids is important in the research and diagnosis of skin diseases. Although punch biopsy is the first-choice technique for the skin sampling, it is unnecessarily invasive for obtaining a sample just for the epidermal analysis. Here we compare two less invasive methods, suction blistering (SB) and tape stripping (TS), for the analysis of selected epidermal genes (quantitative real-time reverse transcription PCR, qRT-PCR), proteins (western blotting, WB), and lipids in ten healthy volunteers...
September 13, 2017: Archives of Dermatological Research
https://www.readbyqxmd.com/read/28904509/peripheral-slc6a4-gene-expression-in-obsessive-compulsive-disorder-in-the-han-chinese-population
#10
Xuemei Wang, Qing Zhao, Wen Chen, Shunying Yu, Zhen Wang, Zeping Xiao
BACKGROUND: Serotonergic system dysfunction has been implicated in obsessive-compulsive disorder (OCD). This study examined peripheral SLC6A4 gene expression in OCD patients and healthy controls to explore the relationship between SLC6A4 and OCD. METHODS: Participants included 50 first episode OCD patients and 60 age and gender-matched healthy controls. Relative SLC6A4 gene expression were examined by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) in peripheral leukocytes of all the subjects...
June 25, 2017: Shanghai Archives of Psychiatry
https://www.readbyqxmd.com/read/28903312/gnp-gapdh1-22-nanovaccines-prevent-neonatal-listeriosis-by-blocking-microglial-apoptosis-and-bacterial-dissemination
#11
Ricardo Calderon-Gonzalez, Elisabet Frande-Cabanes, Hector Teran-Navarro, José María Marimon, Javier Freire, David Salcines-Cuevas, M Carmen Fariñas, Claudia Onzalez-Rico, Marco Marradi, Isabel Garcia, Mirian Alkorta-Gurrutxaga, Aida San Nicolas-Gomez, Ana Castañeda-Sampedro, Sonsoles Yañez-Diaz, Soledad Penades, Carmen Punzon, Javier Gomez-Roman, Fernando Rivera, Manuel Fresno, Carmen Alvarez-Dominguez
Clinical cases of neonatal listeriosis are associated with brain disease and fetal loss due to complications in early or late pregnancy, which suggests that microglial function is altered. This is believed to be the first study to link microglial apoptosis with neonatal listeriosis and listeriosis-associated brain disease, and to propose a new nanovaccine formulation that reverses all effects of listeriosis and confers Listeria monocytogenes (LM)-specific immunity. We examined clinical cases of neonatal listeriosis in 2013-2015 and defined two useful prognostic immune biomarkers to design listeriosis vaccines: high anti-GAPDH1-22 titres and tumor necrosis factor (TNF)/interleukin (IL)-6 ratios...
August 15, 2017: Oncotarget
https://www.readbyqxmd.com/read/28895093/sirna-mediated-rna-interference-in-precision-cut-tissue-slices-prepared-from-mouse-lung-and-kidney
#12
Mitchel J R Ruigrok, Nalinie Maggan, Delphine Willaert, Henderik W Frijlink, Barbro N Melgert, Peter Olinga, Wouter L J Hinrichs
Small interfering RNA (siRNA)-mediated RNAi interference (RNAi) is a powerful post-transcriptional gene silencing mechanism which can be used to study the function of genes in vitro (cell cultures) and in vivo (animal models). However, there is a translational gap between these models. Hence, there is a need for novel experimental models that combine the advantages of in vitro and in vivo models (e.g., simplicity, flexibility, throughput, and representability) to study the effects of siRNA. This need may be addressed by precision-cut tissue slices (PCTS), which represent an ex vivo model that mimics the structural and functional characteristics of a whole organ...
September 11, 2017: AAPS Journal
https://www.readbyqxmd.com/read/28893245/age-dependent-redox-status-in-the-brain-stem-of-no-deficient-hypertensive-rats
#13
Miroslava Majzúnová, Zuzana Pakanová, Peter Kvasnička, Peter Bališ, Soňa Čačányiová, Ima Dovinová
BACKGROUND: The brain stem contains important nuclei that control cardiovascular function via the sympathetic nervous system (SNS), which is strongly influenced by nitric oxide. Its biological activity is also largely determined by oxygen free radicals. Despite many experimental studies, the role of AT1R-NAD(P)H oxidase-superoxide pathway in NO-deficiency is not yet sufficiently clarified. We determined changes in free radical signaling and antioxidant and detoxification response in the brain stem of young and adult Wistar rats during chronic administration of exogenous NO inhibitors...
September 11, 2017: Journal of Biomedical Science
https://www.readbyqxmd.com/read/28889910/phylogenetic-study-of-the-colletotrichum-species-on-imported-citrus-fruits-uncovers-a-low-diversity-and-a-new-species-in-the-colletotrichum-gigasporum-complex
#14
Clovis Douanla-Meli, Jens-Georg Unger
Colletotrichum species associated with citrus fruits are fragmentarily known and it lacks accordingly accurate information on the diversity carried alongside the trade of these commodities from producer countries to Europe. In this study, we investigated the molecular phylogenetic diversity, colonisation, and prevalence of Colletotrichum isolated from asymptomatic and diseased tissues of nine citrus fruit species from 17 geographically diverse countries. Totally 454 isolates were morphoculturally characterised, and multilocus analyses (ACT, ApMat, CHS-1, GAPDH, ITS, TUB2) was performed on a subset of representative morphotype isolates...
October 2017: Fungal Biology
https://www.readbyqxmd.com/read/28889064/spiroplasma-eriocheiris-induces-mouse-3t6-swiss-albino-cell-apoptosis-that-associated-with-the-infection-mechanism
#15
Libo Hou, Wei Gu, Huanxi Zhu, Wei Yao, Wen Wang, Qingguo Meng
Spiroplasma eriocheiris is a novel pathogen similar to the Spiroplasma mirum and also had an ability to infect the newborn mice and caused cataract. Our study was designed to study how S. eriocheiris infects mouse 3T6-Swiss albino cells and to elucidate the cellular molecular pathogenesis of Spiroplasma. FCM analysis and MTT analysis clearly shown that S. eriocheiris could induce 3T6 cell apoptosis and cause cell viability decreased seriously. Immunofluorescence experiments and TEM analysis shown that S. eriocheiris can invade 3T6 cells and form typical inclusion bodies and exhibit vacuolization in vitro...
September 7, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28884171/model-guided-identification-of-novel-gene-amplification-targets-for-improving-succinate-production-in-escherichia-coli-nzn111
#16
Xingxing Jian, Ningchuan Li, Qian Chen, Qiang Hua
Reconstruction and application of genome-scale metabolic models (GEMs) have facilitated metabolic engineering by providing a platform on which systematic computational analysis of metabolic networks can be performed. In this study, a GEM of Escherichia coli NZN111 was employed by the analysis of production and growth coupling (APGC) algorithm to identify genetic strategies for the overproduction of succinate. Through in silico simulation and reaction expression analysis, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoglycerate kinase (PGK), triosephosphate isomerase (TPI), and phosphoenolpyruvate carboxylase (PPC), encoded by gapA, pgk, tpiA, and ppc, respectively, were selected for experimental overexpression...
September 8, 2017: Integrative Biology: Quantitative Biosciences From Nano to Macro
https://www.readbyqxmd.com/read/28880574/selection-of-suitable-internal-control-genes-for-accurate-normalization-of-real-time-quantitative-pcr-data-of-buffalo-bubalus-bubalis-blastocysts-produced-by-scnt-and-ivf
#17
Tanushri Jerath Sood, Swati Viviyan Lagah, Ankita Sharma, Suresh Kumar Singla, Manishi Mukesh, Manmohan Singh Chauhan, Radheysham Manik, Prabhat Palta
We evaluated the suitability of 10 candidate internal control genes (ICGs), belonging to different functional classes, namely ACTB, EEF1A1, GAPDH, HPRT1, HMBS, RPS15, RPS18, RPS23, SDHA, and UBC for normalizing the real-time quantitative polymerase chain reaction (qPCR) data of blastocyst-stage buffalo embryos produced by hand-made cloning and in vitro fertilization (IVF). Total RNA was isolated from three pools, each of cloned and IVF blastocysts (n = 50/pool) for cDNA synthesis. Two different statistical algorithms geNorm and NormFinder were used for evaluating the stability of these genes...
September 7, 2017: Cellular Reprogramming
https://www.readbyqxmd.com/read/28878356/selection-of-reference-genes-suitable-for-normalization-of-qpcr-data-under-abiotic-stresses-in-bioenergy-crop-arundo-donax-l
#18
Michele Poli, Silvio Salvi, Mingai Li, Claudio Varotto
Suitable reference gene selection in qRT-PCR is a key pre-requisite to produce reliable data in gene expression analyses. In this study, novel primers for six commonly used reference genes (AC1, TLF, Act2, TUB α, EF-1α and GAPDH) plus two new candidates (pDUF221 and RPN6) were designed and comparatively tested for expression stability under abiotic stresses (osmotic, heavy metal and heat shock) in shoot, root and their combination of Arundo donax L., a raising non-food energy crop. Expression stability rankings from the most to the least stable gene in each condition and in two tissues (young shoots and roots) were generated with geNorm, NormFinder and BestKeeper programs...
September 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28868355/stability-of-gene-expression-in-human-t-cells-in-different-gravity-environments-is-clustered-in-chromosomal-region-11p15-4
#19
Cora S Thiel, Andreas Huge, Swantje Hauschild, Svantje Tauber, Beatrice A Lauber, Jennifer Polzer, Katrin Paulsen, Hartwin Lier, Frank Engelmann, Burkhard Schmitz, Andreas Schütte, Liliana E Layer, Oliver Ullrich
In the last decades, a plethora of in vitro studies with living human cells contributed a vast amount of knowledge about cellular and molecular effects of microgravity. Previous studies focused mostly on the identification of gravity-responsive genes, whereas a multi-platform analysis at an integrative level, which specifically evaluates the extent and robustness of transcriptional response to an altered gravity environment was not performed so far. Therefore, we investigated the stability of gene expression response in non-activated human Jurkat T lymphocytic cells in different gravity environments through the combination of parabolic flights with a suborbital ballistic rocket and 2D clinostat and centrifuge experiments, using strict controls for excluding all possible other factors of influence...
2017: NPJ Microgravity
https://www.readbyqxmd.com/read/28864008/temporal-and-spatial-expression-of-glyceraldehyde-3-phosphate-dehydrogenase-gapdh-in-the-mouse-placenta
#20
Sarah Min, Bryony V Natale, David R C Natale
Glucose metabolism in trophoblast cells is essential to provide the required energy for the development and function of the placenta. Glyceraldehyde 3-phosphate dehydrogenase (Gapdh), a key enzyme in the glycolysis pathway has been considered ubiquitously expressed in cells. There is, however, a growing body of evidence suggesting that Gapdh has many functions in pathways unrelated to glucose metabolism. In the present study, we show that GAPDH expression and sub-cellular localization changes through gestation in the mouse placenta...
September 2017: Placenta
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