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Manuela Cardi, Mirko Zaffagnini, Alessia De Lillo, Daniela Castiglia, Kamel Chibani, José Manuel Gualberto, Nicolas Rouhier, Jean-Pierre Jacquot, Sergio Esposito
A cDNA coding for a plastidic P2-type G6PDH isoform from poplar (Populus tremula x tremuloides) has been used to express and purify to homogeneity the mature recombinant protein with a N-terminus His-tag. The study of the kinetic properties of the recombinant enzyme showed an in vitro redox sensing modulation exerted by reduced DTT. The interaction with thioredoxins (TRXs) was then investigated. Five cysteine to serine variants (C145S - C175S - C183S - C195S - C242S) and a variant with a double substitution for Cys(175) and Cys(183) (C175S/C183S) have been generated, purified and biochemically characterized in order to investigate the specific role(s) of cysteines in terms of redox regulation and NADPH-dependent inhibition...
November 2016: Plant Science: An International Journal of Experimental Plant Biology
Gro-Ingunn Hemre, Erik-Jan Lock, Pål Asgeir Olsvik, Kristin Hamre, Marit Espe, Bente Elisabeth Torstensen, Joana Silva, Ann-Cecilie Hansen, Rune Waagbø, Johan S Johansen, Monica Sanden, Nini H Sissener
Aiming to re-evaluate current recommendations for nutrient supplementations when Atlantic salmon are fed diets based on plant ingredients, two regression experiments, with parr and post-smolt, were conducted. A control diet was included to evaluate if ingredients supplied sufficient nutrients without any added nutrient package (NP). The nutrient package consisted of vitamins B, C, E, minerals, cholesterol, methionine, taurine and histidine. This paper focus on B-vitamins. In parr, growth, health and welfare parameters responded on NP additions, but this was not observed in the seawater stage...
2016: PeerJ
Mattia Zaccarin, Valentina Bosello-Travain, Maria Luisa Di Paolo, Marco Falda, Matilde Maiorino, Giovanni Miotto, Stefano Piccolo, Antonella Roveri, Fulvio Ursini, Rina Venerando, Stefano Toppo
Reversible oxidation of Cys residues is a crucial element of redox homeostasis and signaling. According to a popular concept in oxidative stress signaling, the oxidation of targets of signals can only take place following an overwhelming of the cellular antioxidant capacity. This concept, however, ignores the activation of feedback mechanisms possibly leading to a paradoxical effect. In a model of cancer stem cells (CSC), stably overexpressing the TAZ oncogene, we observed that the increased formation of oxidants is associated with a globally more reduced state of proteins...
September 13, 2016: Archives of Biochemistry and Biophysics
Cristina Salmerón, Natàlia Riera-Heredia, Joaquim Gutiérrez, Isabel Navarro, Encarnación Capilla
During the last decades, adipogenesis has become an emerging field of study in aquaculture due to the relevance of the adipose tissue in many physiological processes and its connection with the endocrine system. In this sense, recent studies have translated into the establishment of preadipocyte culture models from several fish species, sometimes lacking information on the mRNA levels of adipogenic genes. Thus, the aim of this study was to determine the gene expression profile of gilthead sea bream (Sparus aurata) primary cultured mesenchymal stem cells (MSCs) from different origin (adipose tissue and vertebra bone) during adipogenesis...
2016: Frontiers in Endocrinology
Mahbobeh Eshtiyaghi, Hamid Deldar, Zarbakht Ansari Pirsaraei, Bahram Shohreh
The aim of this study was to investigate the effect of different concentrations of royal jelly (RJ) on in vitro maturation (IVM), fertilization, cleavage, blastocyst rates, glutathione (GSH) content in ovine oocyte, mRNA abundance of antioxidant enzymes in both oocyte and cumulus, and glucose metabolism-related genes in cumulus cells. In vitro maturation of oocyte was performed in the presence of control (RJ0), 2.5 (RJ2.5), 5 (RJ5), and 10 (RJ10) mg/mL of RJ. Nuclear status, intracellular GSH content in oocytes, and mRNA abundance of selected genes were evaluated following 24 hours of IVM...
July 25, 2016: Theriogenology
Rychelle Clayde Affonso Medeiros, Karina Girardi do Carmo de Vasconcelos, Fernanda Karlla Luz Cardoso, Bruno de Siqueira Mieto, Thiago Gomes de Toledo Pinto, Lilian Sales Gomez, Luciana Silva Rodrigues, Mariana Gandini, Julio Jablonski Amaral, Sérgio Luiz Gomes Antunes, Suzana Corte-Real, Patricia Sammarco Rosa, Maria Cristina Vidal Pessolani, José Augusto da Costa Nery, Euzenir Nunes Sarno, Leonardo Ribeiro Batista Silva, Mauro Sola-Penna, Marcus Fernandes de Oliveira, Milton Ozório Moraes, Flavio Alves Lara
Mycobacterium leprae, the intracellular etiological agent of leprosy, infects Schwann promoting irreversible physical disabilities and deformities. These cells are responsible for myelination and maintenance of axonal energy metabolism through export of metabolites, such as lactate and pyruvate. In the present work, we observed that infected Schwann cells increase glucose uptake with a concomitant increase in glucose-6-phosphate dehydrogenase (G6PDH) activity, the key enzyme of the oxidative pentose pathway...
August 23, 2016: Journal of Biological Chemistry
Amalia Pérez-Jiménez, Emilia Abellán, Marta Arizcun, Gabriel Cardenete, Amalia E Morales, M Carmen Hidalgo
Common dentex (Dentex dentex) is an appreciated carnivorous fish with high growth rate and life cycle adaptable to existing farming techniques. Since the use of carbohydrates is an economic and sustainable alternative for a protein-sparing effect, the study of how this macronutrient affects the welfare of carnivorous species must be studied. The aim of the present study was to evaluate the effects of different types and levels of carbohydrates on common dentex oxidative status. Nine isonitrogenous (43%) and isoenergetic (22MJkg(-1)) diets were formulated combining three types (pregelatinized starch-PS, dextrin-Dx and maltodextrin-Mx) and three levels (12, 18 and 24%) of carbohydrates...
August 21, 2016: Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology
Franziska Visser, Boje Müller, Judith Rose, Dirk Prüfer, Gundula A Noll
The immobilisation of enzymes plays an important role in many applications, including biosensors that require enzyme activity, stability and recyclability in order to function efficiently. Here we show that forisomes (plant-derived mechanoproteins) can be functionalised with enzymes by translational fusion, leading to the assembly of structures designated as forizymes. When forizymes are expressed in the yeast Saccharomyces cerevisiae, the enzymes are immobilised by the self-assembly of forisome subunits to form well-structured protein bodies...
2016: Scientific Reports
Baoliang Liu, Rui Jia, Cen Han, Bin Huang, Ji-Lin Lei
This study was designed to evaluate the physiological and immune responses of juvenile turbot to stocking density. Turbot (average weight 185.4g) were reared for 120days in a land based recirculating aquaculture system (RAS) under three stocking densities: low density (LD, ~9.3-26.1kg/m(2), initial to final density), medium density (MD, ~13.6-38.2kg/m(2)) and high density (HD, ~19.1-52.3kg/m(2)). Fish were sampled at days 0, 40, 80 and 120 to obtain growth parameters and liver tissues. No significant difference was detected in growth, biochemical parameters and gene expression among the three densities until at the final sampling (day 120)...
August 3, 2016: Comparative Biochemistry and Physiology. Toxicology & Pharmacology: CBP
Moysés S Miranda, Hamilton S Nascimento, Mayra P R Costa, Nathália N Costa, Karynne N L Brito, Cinthia T A Lopes, Simone S D Santos, Marcela S Cordeiro, Otávio M Ohashi
PURPOSE: Despite advances in the composition of defined embryo culture media, co-culture with somatic cells is still used for bovine in vitro embryo production (IVEP) in many laboratories worldwide. Granulosa cells are most often used for this purpose, although recent work suggests that co-culture with stem cells of adult or embryonic origin or their derived biomaterials may improve mouse, cattle, and pig embryo development. MATERIALS AND METHODS: In experiment 1, in vitro produced bovine embryos were co-cultured in the presence of two concentrations of bovine adipose tissue-derived mesenchymal cells (b-ATMSCs; 10(3) and 10(4) cells/mL), in b-ATMSC preconditioned medium (SOF-Cond), or SOF alone (control)...
October 2016: Journal of Assisted Reproduction and Genetics
Guiqing Liu, Xuehong Qiu, Li Cao, Yi Zhang, Zubing Zhan, Richou Han
Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) is the sensitive method to quantify the expression levels of target genes on the basis of endogenous control. An appropriate reference gene set for normalization is essential for reliable results. The ghost moth, Thitarodes armoricanus, a host species of a medicinal fungus, Ophiocordyceps sinensis, is an economically important member of the Lepidoptera. Recent studies have focused on the mechanism of adaptation of this species to its high-altitude environment and host immune response to O...
2016: PloS One
Mehrdad Ghashghaeinia, Daniela Giustarini, Pavla Koralkova, Martin Köberle, Kousi Alzoubi, Rosi Bissinger, Zohreh Hosseinzadeh, Peter Dreischer, Ingolf Bernhardt, Florian Lang, Mahmoud Toulany, Thomas Wieder, Renata Mojzikova, Ranieri Rossi, Ulrich Mrowietz
In mature erythrocytes, glucose-6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconate dehydrogenase (6PGDH) yield NADPH, a crucial cofactor of the enzyme glutathione reductase (GR) converting glutathione disulfide (GSSG) into its reduced state (GSH). GSH is essential for detoxification processes in and survival of erythrocytes. We explored whether the anti-inflammatory compounds Bay 11-7082, parthenolide and dimethyl fumarate (DMF) were able to completely deplete a common target (GSH), and to impair the function of upstream enzymes of GSH recycling and replenishment...
2016: Scientific Reports
Julie Grasset, Élodie Ollivier, Bérénice Bougas, Glenn Yannic, Peter G C Campbell, Louis Bernatchez, Patrice Couture
In this study, we measured the effects of temperature (9°C, 20°C, and 28°C), metal contamination (cadmium and nickel) and their interaction on yellow perch (Perca flavescens) using liver enzymatic and transcriptomic endpoints and biometric indices. Kidney metal concentrations increased with a rise of temperature. The biometric indices analysed (Fulton condition factor, pyloric cæca, hepatosomatic and gonadosomatic indices) generally decreased with an increase of temperature but not with metal contamination...
August 2016: Aquatic Toxicology
Roger W L Godschalk, Kimberly Vanhees, Lou Maas, Marie-Jose Drittij, Daniëlle Pachen, Sahar van Waalwijk van Doorn-Khosrovani, Frederik J van Schooten, Guido R M M Haenen
A balanced redox homeostasis in the testis is essential for genetic integrity of sperm. Reactive oxygen species can disturb this balance by oxidation of glutathione, which is regenerated using NADPH, formed by glucose-6-phosphate dehydrogenase (G6PDH). G6PDH is regulated by the Ataxia Telangiectasia Mutated (Atm) protein. Therefore, we studied the redox status and DNA damage in testes and sperm of mice that carried a deletion in Atm. The redox status in heterozygote mice, reflected by glutathione levels and antioxidant capacity, was lower than in wild type mice, and in homozygotes the redox status was even lower...
August 2016: Reproductive Toxicology
Rômi S Piazza, Rafael Trevisan, Fabrício Flores-Nunes, Guilherme Toledo-Silva, Nestor Wendt, Jacó J Mattos, Daína Lima, Satie Taniguchi, Silvio Tarou Sasaki, Álvaro C P Mello, Flávia L Zacchi, Miguel A S Serrano, Carlos H A M Gomes, Márcia C Bícego, Eduardo A de Almeida, Afonso C D Bainy
Understanding the mechanism of phenanthrene (PHE) biotransformation and related cellular responses in bivalves can be an important tool to elucidate the risks of polycyclic aromatic hydrocarbons (PAHs) to aquatic organisms. In the present study it was analyzed the transcriptional levels of 13 biotransformation genes related to cytochrome P450 (CYP), glutathione S-transferase (GST), sulfotransferase (SULT), flavin-containing monooxygenase and fatty acid-binding proteins by qPCR in gill of scallops Nodipecten nodosus exposed for 24 or 96h to 50 or 200μgL(-1) PHE (equivalent to 0...
August 2016: Aquatic Toxicology
Huahua Wang, Lidan Yang, Yan Li, Junjie Hou, Junjun Huang, Weihong Liang
The roles of abscisic acid (ABA) and hydrogen peroxide (H2O2) in inducing glucose-6-phosphate dehydrogenase (G6PDH, EC activity and the possible roles of G6PDH in regulating ascorbate-glutathione (AsA-GSH) cycle were investigated in soybean (Glycine max L.) roots under drought stress. Drought caused a marked increase of the total and cytosolic G6PDH activities and triggered a rapid ABA and H2O2 accumulation in soybean roots. Exogenous ABA or H2O2 treatment elevated the total and cytosolic G6PDH activities, whereas suppressing ABA or H2O2 production inhibited the drought-induced increase in total and cytosolic G6PDH activities, suggesting that ABA and H2O2 are required for drought-induced increase of total G6PDH activity, namely cytosolic G6PDH activity...
October 2016: Plant Physiology and Biochemistry: PPB
Joanna Jakubowska, Agnieszka Maciejewska, Ryszard Pawłowski
Recently mRNA profiling has been widely proposed as a universal tool for biological fluids identification. Here, we describe a test for vaginal fluid identification that combines detection of five markers: vaginal mRNAs and Lactobacilli in end-point PCR reaction. The test detects the following transcripts: HBD1 (Human beta-defensin 1), MUC4 (Mucin 4), MMP11 (Matrix metalloproteinase 11), housekeeping gene G6PDH (glucose 6-phosphate dehydrogenase), and the 16S-23S rRNA intergenic spacer regions of L. crispatus and L...
2016: Methods in Molecular Biology
Ivan Viegas, Ivana Jarak, João Rito, Rui A Carvalho, Isidoro Metón, Miguel A Pardal, Isabel V Baanante, John G Jones
Farmed seabass have higher adiposity than their wild counterparts and this is often attributed to carbohydrate (CHO) feeding. Whether this reflects a reduction in fat oxidation, increased de novo lipogenesis (DNL), or both, is not known. To study the effects of high CHO diets on hepatic TG biosynthesis, hepatic TG deuterium ((2)H) enrichment was determined following 6 days in (2)H-enriched tank water for fish fed with a no-CHO control diet (CTRL), and diets with digestible starch (DS) and raw starch (RS). Hepatic fractional synthetic rates (FSRs, percent per day(-1)) were calculated for hepatic TG-glyceryl and FA moieties through (2)H NMR analysis...
July 2016: Journal of Lipid Research
Wei Li, Bo Li, Jiaqi Lv, Li Dong, Lili Zhang, Tian Wang
The objective of this study was to investigate the effects of dietary choline supplementation on hepatic lipid metabolism and gene expression in finishing pigs with intrauterine growth retardation (IUGR). Using a 2 × 2 factorial design, eight normal birth weight (NBW) and eight IUGR weaned pigs were fed either a basal diet (NBW pigs fed a basal diet, NC; IUGR pigs fed a basal diet, IC) or a diet supplemented with two times more choline than the basal diet (NBW pigs fed a high-choline diet, NH; IUGR pigs fed a high-choline diet, IH) until 200 d of age...
May 22, 2016: Asian-Australasian Journal of Animal Sciences
Neeraj Kumar, K Ambasankar, K K Krishnani, S K Gupta, Shashi Bhushan, P S Minhas
This study investigated 96h median lethal concentration of endosulfan (99%, pure α: β ratio of 7:3) by conducting static non-renewable acute toxicity bio-assay in Chanos chanos juvenile with average weight (110±5.65g). Further, the effect of different definitive doses (18.5, 19.5, 20.5, 21.5 and 22.5µg/L) of endosulfan on metabolic, heamato-immunoligcal and histopathological response were probed. Anti-oxidative enzymes CAT, SOD and GST showed significant (p<0.01) increase of activity in the liver, gill and brain during exposure to endosulfan in a dose and time dependent manner...
September 2016: Ecotoxicology and Environmental Safety
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