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https://www.readbyqxmd.com/read/28642848/mycobacterium-tuberculosis-glyceraldehyde-3-phosphate-dehydrogenase-gapdh-functions-as-a-receptor-for-human-lactoferrin
#1
Himanshu Malhotra, Anil Patidar, Vishant M Boradia, Rajender Kumar, Rakesh D Nimbalkar, Ajay Kumar, Zahid Gani, Rajbeer Kaur, Prabha Garg, Manoj Raje, Chaaya I Raje
Iron is crucial for the survival of living cells, particularly the human pathogen Mycobacterium tuberculosis (M.tb) which uses multiple strategies to acquire and store iron. M.tb synthesizes high affinity iron chelators (siderophores), these extract iron from host iron carrier proteins such as transferrin (Tf) and lactoferrin (Lf). Recent studies have revealed that M.tb may also relocate several housekeeping proteins to the cell surface for capture and internalization of host iron carrier protein transferrin...
2017: Frontiers in Cellular and Infection Microbiology
https://www.readbyqxmd.com/read/28635628/identification-and-validation-of-reference-genes-for-seashore-paspalum-response-to-abiotic-stresses
#2
Yu Liu, Jun Liu, Lei Xu, Hui Lai, Yu Chen, Zhimin Yang, Bingru Huang
Seashore paspalum (Paspalum vaginatum) is among the most salt- and cadmium-tolerant warm-season perennial grass species widely used as turf or forage. The objective of this study was to select stable reference genes for quantitative real-time polymerase chain reaction (qRT-PCR) analysis of seashore paspalum in response to four abiotic stresses. The stability of 12 potential reference genes was evaluated by four programs (geNorm, NormFinder, BestKeeper, and RefFinder). U2AF combined with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) showed stable expression in Cd-treated leaves and cold-treated roots...
June 21, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28631898/mass-spectrometry-based-protein-analytics-to-unravel-the-tissue-pathophysiology-in-duchenne-muscular-dystrophy
#3
Stephanie J Carr, René P Zahedi, Hanns Lochmüller, Andreas Roos
Duchenne muscular dystrophy (DMD) is a genetic muscle wasting condition with limited treatment options available and is caused by the lack of Dystrophin. However, pathophysiology of different tissues is variable showing different histological and molecular signatures. Recently, a number of studies have employed gel-free proteomic approaches to unveil the molecular pathophysiology in terms of tissue-specific proteome changes in dystrophin-deficiency. We analysed studies in models of dystrophin-deficiency and patients both from the published literature...
June 20, 2017: Proteomics. Clinical Applications
https://www.readbyqxmd.com/read/28625912/isolation-of-recombinant-human-untagged-glyceraldehyde-3-phosphate-dehydrogenase-from-e-%C3%A2-coli-producer-strain
#4
K V Barinova, M A Eldarov, E V Khomyakova, V I Muronetz, E V Schmalhausen
The goal of the present work was expression of human glyceraldehyde-3-phosphate dehydrogenase (hGAPDH) without additional tag constructions in E. coli cells and elaboration of the procedure for purification of untagged hGAPDH from the extract of the producer cells. We present a simple method for purification of untagged hGAPDH including ammonium sulfate fractionation and gel filtration on a G-100 Sephadex column. The method allows isolation of 2 mg of pure hGAPDH from 600 ml of cell culture (7 g of the cell biomass)...
June 15, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28625896/lateralization-of-housekeeping-genes-in-the-brain-of-one-day-old-chicks
#5
Jacek Lenart, Klaudia Kogut, Elzbieta Salinska
Real-time quantitative PCR is an exceptionally sensitive method that can detect even very small differences in gene expression and, as such, it is essential to use suitable reference genes. Domestic chickens are used in a wide range of studies including neurobiology, behavior, ecology and disease transmission. In recent avian gene expression experiments, 18S (18S ribosomal RNA), beta actin (ACTB) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) have frequently been used; however, there is not enough evidence that these reference genes are suitable for all types of experiments...
June 15, 2017: Gene Expression Patterns: GEP
https://www.readbyqxmd.com/read/28625264/-suppressive-effect-of-corm-2-on-platelet-%C3%AE-granule-exocytosis-in-sepsis-via-snare-munc18b-complex-formation
#6
Mingfeng Zhuang, Bingwei Sun, Dadong Liu, Yuan Shi
OBJECTIVE: To investigate the suppressive effect of carbon monoxide-releasing molecule II (CORM-2) on LPS induced platelet α-granule exocytosis in sepsis via soluble N-ethylmaleimide-sensitive factor attached protein receptor/mammalian uncoordinated 18b (SNARE/Munc18b) complex formation. METHODS: Blood was collected from healthy volunteers' cubital vein, then platelets were isolated by differential centrifugation. Platelets were randomly divided into 5 groups. The control group did not undergo any treatment, the LPS group received 10 mg/L LPS simulation, the CORM-2 group and iCORM-2 group underwent LPS simulation and immediate administration of CORM-2 (10 μmol/L and 50 μmol/L) or iCORM-2 (50 μmol/L), respectively...
February 2017: Zhonghua Wei Zhong Bing Ji Jiu Yi Xue
https://www.readbyqxmd.com/read/28624280/technical-note-evaluation-of-endogenous-control-gene-expression-in-bovine-neutrophils-by-reverse-transcription-quantitative-pcr-using-microfluidics-gene-expression-arrays
#7
M A Crookenden, C G Walker, B Kuhn-Sherlock, A Murray, V S R Dukkipati, A Heiser, J R Roche
Reverse-transcription quantitative-PCR (RT-qPCR) is commonly used for assessing the cellular response to changes in physiologic and pathologic conditions. The selection of stable endogenous control genes is an important step of any RT-qPCR study, as expression can vary depending on the experimental environment. Our objective was to identify endogenous control genes in circulating neutrophils isolated from cows during the peripartum period. To do this, we used microfluidics gene expression arrays (Fluidigm, San Francisco, CA) for RT-qPCR analysis...
June 14, 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/28623342/hif-1%C3%AE-knockdown-reduces-glycolytic-metabolism-and-induces-cell-death-of-human-synovial-fibroblasts-under-normoxic-conditions
#8
Manuel J Del Rey, Álvaro Valín, Alicia Usategui, Carmen M García-Herrero, María Sánchez-Aragó, José M Cuezva, María Galindo, Beatriz Bravo, Juan D Cañete, Francisco J Blanco, Gabriel Criado, José L Pablos
Increased glycolysis and HIF-1α activity are characteristics of cells under hypoxic or inflammatory conditions. Besides, in normal O2 environments, elevated rates of glycolysis support critical cellular mechanisms such as cell survival. The purpose of this study was to analyze the contribution of HIF-1α to the energy metabolism and survival of human synovial fibroblasts (SF) under normoxic conditions. HIF-1α was silenced using lentiviral vectors or small-interfering RNA (siRNA) duplexes. Expression analysis by qRT-PCR and western blot of known HIF-1α target genes in hypoxia demonstrated the presence of functional HIF-1α in normoxic SF and confirmed the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a HIF-1α target even in normoxia...
June 16, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28612110/differential-killing-and-radio-modifying-effects-of-iodoacetate-in-mammalian-normal-and-cancer-cells
#9
Usha Yadav, K B Anjaria, Rajesha Nairy, K B Shirsath, Utkarsha N Desai, Rajesh K Chaurasia, Nagesh N Bhat, B K Sapra
To explore possible applications of iodoacetate (IA), a glycolytic inhibitor, in cancer treatment, we screened its cytotoxicity and radioprotective/sensitizing efficacy in three different mammalian cell lines; A549 (human lung carcinoma), MCF7 (human mammary cancer), a non-cancerous CHO (Chinese hamster ovary) cells and human lymphocytes. Experiments were carried out using IA concentrations ranging from 0.01 to 2.5 µg/ml, with or without (60)Coγ-radiation. In the outcomes, IA was found to exhibit higher toxicity in the cancer cells, whereas it was non-toxic/marginally toxic to the non-cancerous cells...
June 13, 2017: Radiation and Environmental Biophysics
https://www.readbyqxmd.com/read/28609925/-i-epirrita-i-i-pulchraria-i-taylor-1907-transferred-to-i-malacodea-i-with-notes-on-the-phylogeny-and-ecology-of-the-tribe-operophterini-lepidoptera-geometridae-larentiinae
#10
Andro Truuverk, Erki Õunap, Toomas Tammaru
The Nearctic Epirrita pulchraria (Taylor, 1907) was revealed as a sister taxon of the Palaearctic Malacodea regelaria Tengström,1869 in a molecular phylogenetic analysis of the tribe Operophterini. DNA sequence variation in one mitochondrial (COI) and eight nuclear (28S, EF-1α, WGL, GAPDH, RPS5, IDH, MDH, CAD) gene fragments was used in the analysis. Bayesian inference resulted in a well-resolved phylogenetic hypothesis. According to the recovered phylogeny a new combination is proposed: Rachela pulchraria Taylor, 1907 is transferred from Epirrita Hübner, 1808 to Malacodea Tengström, 1869 as Malacodea pulchraria (Taylor, 1907) comb...
April 27, 2017: Zootaxa
https://www.readbyqxmd.com/read/28608814/predicting-outcome-and-therapy-response-in-mcrc-patients-using-an-indirect-method-for-ctcs-detection-by-a-multigene-expression-panel-a-multicentric-prospective-validation-study
#11
Yolanda Vidal Insua, Juan de la Cámara, Elena Brozos Vázquez, Ana Fernández, Francisca Vázquez Rivera, Mª José Villanueva Villanueva Silva, Jorge Barbazán, Laura Muinelo-Romay, Sonia Candamio Folgar, Alicia Abalo, Rafael López-López, Miguel Abal, Lorena Alonso-Alconada
Colorectal cancer (CRC) is one of the major causes of cancer-related deaths. Early detection of tumor relapse is crucial for determining the most appropriate therapeutic management. In clinical practice, computed tomography (CT) is routinely used, but small tumor changes are difficult to visualize, and reliable blood-based prognostic and monitoring biomarkers are urgently needed. The aim of this study was to prospectively validate a gene expression panel (composed of GAPDH, VIL1, CLU, TIMP1, TLN1, LOXL3 and ZEB2) for detecting circulating tumor cells (CTCs) as prognostic and predictive tool in blood samples from 94 metastatic CRC (mCRC) patients...
June 13, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28608282/poly-adp-ribosylated-proteins-in-mononuclear-cells-from-patients-with-type-2-diabetes-identified-by-proteomic-studies
#12
Alessandra Giorgi, Italo Tempera, Giorgia Napoletani, Diego Drovandi, Cinzia Potestà, Sara Martire, Elisabetta Mandosi, Tiziana Filardi, M Eugenia Schininà, Susanna Morano, Maria d'Erme, Bruno Maras
AIMS: In diabetes, hyperglycemia increases reactive oxygen species that induce DNA damage and poly(ADP-ribose)polymerase activation. The aim of this study is to characterize the proteomic profile and the role of poly(ADP-ribosylation) in patients with type 2 diabetes. METHODS: A proteomic platform based on 2DE and MALDI-ToF spectrometry was applied to peripheral blood mononuclear cells obtained from two different cohorts in which diabetic (n = 14) and normoglycemic patients (n = 11) were enrolled...
June 12, 2017: Acta Diabetologica
https://www.readbyqxmd.com/read/28607089/weak-protein-protein-interactions-in-live-cells-are-quantified-by-cell-volume-modulation
#13
Shahar Sukenik, Pin Ren, Martin Gruebele
Weakly bound protein complexes play a crucial role in metabolic, regulatory, and signaling pathways, due in part to the high tunability of their bound and unbound populations. This tunability makes weak binding (micromolar to millimolar dissociation constants) difficult to quantify under biologically relevant conditions. Here, we use rapid perturbation of cell volume to modulate the concentration of weakly bound protein complexes, allowing us to detect their dissociation constant and stoichiometry directly inside the cell...
June 12, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28601074/role-of-glyceraldehyde-3-phosphate-dehydrogenase-gapdh-in-dna-repair
#14
REVIEW
A A Kosova, S N Khodyreva, O I Lavrik
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is widely known as a glycolytic enzyme. Nevertheless, various functions of GAPDH have been found that are unrelated to glycolysis. Some of these functions presume interaction of GAPDH with DNA, but the mechanism of its translocation to the nucleus is not fully understood. When in the nucleus, GAPDH participates in the initiation of apoptosis and transcription of genes involved in antiapoptotic pathways and cell proliferation and plays a role in the regulation of telomere length...
June 2017: Biochemistry. Biokhimii︠a︡
https://www.readbyqxmd.com/read/28598997/selection-of-reference-genes-for-quantitative-real-time-rt-pcr-assays-in-different-morphological-forms-of-dimorphic-zygomycetous-fungus-benjaminiella-poitrasii
#15
Ejaj K Pathan, Vandana Ghormade, Mukund V Deshpande
Benjaminiella poitrasii, a dimorphic non-pathogenic zygomycetous fungus, exhibits a morphological yeast (Y) to hypha (H) reversible transition in the vegetative phase, sporangiospores (S) in the asexual phase and zygospores (Z) in the sexual phase. To study the gene expression across these diverse morphological forms, suitable reference genes are required. In the present study, 13 genes viz. ACT, 18S rRNA, eEF1α, eEF-Tu,eIF-1A, Tub-α, Tub-b, Ubc, GAPDH, Try, WS-21, NADGDH and NADPGDH were evaluated for their potential as a reference, particularly for studying gene expression during the Y-H reversible transition and also for other asexual and sexual life stages of B...
2017: PloS One
https://www.readbyqxmd.com/read/28592345/the-effect-of-kinetic-heat-shock-on-bovine-oocyte-maturation-and-subsequent-gene-expression-of-targeted-genes
#16
Krishna C Pavani, António Rocha, Erica Baron, Joana Lourenço, Marwa Faheem, Fernando Moreira da Silva
The exposure of oocytes to heat stress during the maturation process results in harmful effects to their internal organelles, low fertilization capability and higher embryonic losses. In the present experiment the effect of heat shock (HS) during the maturation process was assessed. In Assay 1, oocytes from winter (December-March; n = 100) and summer (June-September; n = 100) months were collected and matured to analyse their HS tolerance. Total RNA was extracted from matured oocytes and cDNA synthesis was performed, followed by qPCR for selected genes (Cx43, CDH1, DNMT1, HSPA14), compared with two reference genes (GAPDH and SDHA)...
June 8, 2017: Zygote: the Biology of Gametes and Early Embryos
https://www.readbyqxmd.com/read/28591185/selection-of-suitable-endogenous-reference-genes-for-qpcr-in-kidney-and-hypothalamus-of-rats-under-testosterone-influence
#17
Khadijeh Gholami, Su Yi Loh, Naguib Salleh, Sau Kuen Lam, See Ziau Hoe
Real-time quantitative PCR (qPCR) is the most reliable and accurate technique for analyses of gene expression. Endogenous reference genes are being used to normalize qPCR data even though their expression may vary under different conditions and in different tissues. Nonetheless, verification of expression of reference genes in selected studied tissue is essential in order to accurately assess the level of expression of target genes of interest. Therefore, in this study, we attempted to examine six commonly used reference genes in order to identify the gene being expressed most constantly under the influence of testosterone in the kidneys and hypothalamus...
2017: PloS One
https://www.readbyqxmd.com/read/28589519/hydrogel-environment-supports-cell-culture-expansion-of-a-grade-iv-astrocytoma
#18
Manasi P Jogalekar, Leigh G Cooper, Elba E Serrano
Malignant astrocytomas are aggressive cancers of glial origin that can develop into invasive brain tumors. The disease has poor prognosis and high recurrence rate. Astrocytoma cell lines of human origin are an important tool in the experimental pathway from bench to bedside because they afford a convenient intermediate system for in vitro analysis of brain cancer pathogenesis and treatment options. We undertook the current study to determine whether hydrogel culture methods could be adapted to support the growth of astrocytoma cell lines, thereby facilitating a system that may be biologically more similar to in vivo tumor tissue...
June 7, 2017: Neurochemical Research
https://www.readbyqxmd.com/read/28587729/glycolysis-related-proteins-are-broad-spectrum-vaccine-candidates-against-aquacultural-pathogens
#19
Xiaohong Liu, Jiamin Sun, Haizhen Wu
Reverse vaccinology (RV) has become a popular method for developing vaccines. Although Edwardsiella tarda is deemed to be an important fish pathogen, so far, no reports have used a genome-based approach to screen vaccine candidates against E. tarda. In the current study, protective antigens of E. tarda were screened using RV. Large-scale cloning, expression and purification of potential candidates were carried out, and their immunoprotective potential was evaluated. A candidate fructose-bisphosphate aldolase (FBA) exhibited broad spectrum protection, as did another glycolysis-related protein glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which we reported previously, indicating the potential of other glycolysis-related proteins of E...
June 3, 2017: Vaccine
https://www.readbyqxmd.com/read/28584723/selection-of-housekeeping-genes-as-internal-controls-for-quantitative-rt-pcr-analysis-of-the-veined-rapa-whelk-rapana-venosa
#20
Hao Song, Xin Dang, Yuan-Qiu He, Tao Zhang, Hai-Yan Wang
BACKGROUND: The veined rapa whelk Rapana venosa is an important commercial shellfish in China and quantitative real-time PCR (qRT-PCR) has become the standard method to study gene expression in R. venosa. For accurate and reliable gene expression results, qRT-PCR assays require housekeeping genes as internal controls, which display highly uniform expression in different tissues or stages of development. However, to date no studies have validated housekeeping genes in R. venosa for use as internal controls for qRT-PCR...
2017: PeerJ
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