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https://www.readbyqxmd.com/read/29341594/designing-flavoprotein-gfp-fusion-probes-for-analyte-specific-ratiometric-fluorescence-imaging
#1
Devin A Hudson, Jeffrey L Caplan, Colin Thorpe
The development of genetically encoded fluorescent probes for analyte-specific imaging has revolutionized our understanding of intracellular processes. Current classes of intracellular probes depend on the selection of binding domains that either undergo conformational changes on analyte binding or can be linked to thiol redox chemistry. Here we have designed novel probes by fusing a flavoenzyme, whose fluorescence is quenched on reduction by the analyte of interest, with a GFP domain to allow for rapid and specific ratiometric sensing...
January 17, 2018: Biochemistry
https://www.readbyqxmd.com/read/29338715/landscape-genomics-natural-selection-drives-the-evolution-of-mitogenome-in-penguins
#2
Barbara Ramos, Daniel González-Acuña, David E Loyola, Warren E Johnson, Patricia G Parker, Melanie Massaro, Gisele P M Dantas, Marcelo D Miranda, Juliana A Vianna
BACKGROUND: Mitochondria play a key role in the balance of energy and heat production, and therefore the mitochondrial genome is under natural selection by environmental temperature and food availability, since starvation can generate more efficient coupling of energy production. However, selection over mitochondrial DNA (mtDNA) genes has usually been evaluated at the population level. We sequenced by NGS 12 mitogenomes and with four published genomes, assessed genetic variation in ten penguin species distributed from the equator to Antarctica...
January 16, 2018: BMC Genomics
https://www.readbyqxmd.com/read/29337692/generation-of-superoxide-and-hydrogen-peroxide-by-side-reactions-of-mitochondrial-2-oxoacid-dehydrogenase-complexes-in-isolation-and-in-cells
#3
Victoria I Bunik, Martin D Brand
Mitochondrial 2-oxoacid dehydrogenase complexes oxidize 2-oxoglutarate, pyruvate, branched-chain 2-oxoacids and 2-oxoadipate to the corresponding acyl-CoAs and reduce NAD+ to NADH. The isolated enzyme complexes generate superoxide anion radical or hydrogen peroxide in defined reactions by leaking electrons to oxygen. Studies using isolated mitochondria in media mimicking cytosol suggest that the 2-oxoacid dehydrogenase complexes contribute little to the production of superoxide or hydrogen peroxide relative to other mitochondrial sites at physiological steady-states...
June 27, 2017: Biological Chemistry
https://www.readbyqxmd.com/read/29336483/glucose-metabolism-during-in-vitro-maturation-of-mouse-oocytes-an-study-using-rna-interference
#4
Hong-Li Xie, Shuai Zhu, Jie Zhang, Jing Wen, Hong-Jie Yuan, Liu-Zhu Pan, Ming-Jiu Luo, Jing-He Tan
In previous studies on glucose metabolism during in vitro maturation, intact cumulus-oocyte complexes (COCs) were treated with enzyme inhibitors/activators. Because inhibitors/activators may have non-specificity and/or toxicity, and culture of COCs cannot differentiate whether glucose metabolism of cumulus cells (CCs) or that of the oocyte supports oocyte maturation, results from the previous studies must be verified by silencing genes in either CCs or cumulus-denuded oocytes (DOs). In this study, RNAi was adopted to specify the effects of glucose metabolism in CCs or DOs on oocyte maturation...
January 16, 2018: Journal of Cellular Physiology
https://www.readbyqxmd.com/read/29333085/next-generation-sequencing-expression-profiling-of-mitochondrial-subunits-in-men-with-klinefelter-syndrome
#5
Michele Salemi, Laura Cimino, Marika Marino, Rossella Cannarella, Rosita A Condorelli, Corrado Romano, Sandro La Vignera, Aldo E Calogero
Objectives: Klinefelter syndrome (KS) is one of the most common sex-chromosome disorders as it affects up to 1 in every 600-1000 newborn males. Men with KS carry one extra X chromosome and they usually present a 47,XXY karyotype, but less frequent variants have also been reported in literature. KS typical symptoms include tall stature, gynecomastia, broad hips, hypogonadism and absent spermatogenesis. The syndrome is also related to a wide range of cognitive deficits, among which language-based learning disabilities and verbal cognition impairment are frequently diagnosed...
2018: International Journal of Medical Sciences
https://www.readbyqxmd.com/read/29330287/targeted-akt-inhibition-in-prostate-cancer-cells-and-spheroids-reduces-aerobic-glycolysis-and-generation-of-hyperpolarized-1-13c-lactate
#6
Sui Seng Tee, Izabela Suster, Steven Truong, Sangmoo Jeong, Roozbeh Eskandari, Valentina Di Gialleonardo, Julio A Alvarez, Hannah N Aldeborgh, Kayvan Keshari
The PI3K/AKT/mTOR (PAM) signaling pathway is frequently mutated in prostate cancer. Specific AKT inhibitors are now in advanced clinical trials and this study investigates the effect of MK2206, a non-ATP competitive inhibitor, on the cellular metabolism in the context of prostate cancer. A significant reduction in cell motility and aerobic glycolysis was observed in prostate cancer cells with treatment. These changes were not accompanied by a reduction in the ratio of high-energy phosphates or a change in total protein levels of enzymes (e...
January 12, 2018: Molecular Cancer Research: MCR
https://www.readbyqxmd.com/read/29327817/ketonization-of-proline-residues-in-the-peptide-chains-of-actinomycins-by-a-4-oxoproline-synthase
#7
Siamak Semsary, Ivana Crnovčić, Ronja Driller, Joachim Vater, Bernhard Loll, Ullrich Keller
X-type actinomycins (Acms) contain 4-hydroxyproline (Acm X0) or 4-oxoproline (Acm X2) in their ß-pentapeptide lactone rings whereas their α-ring contains proline. We demonstrate that these Acms are formed through asymmetric condensation of Acm half molecules (Acm halves) containing proline with 4-hydroxyproline- or 4-oxo-proline-containing Acm halves. In turn we show - using an artificial Acm half analogue (PPL 1) with proline in its peptide chain - its conversion into the 4-hydroxyproline and 4-oxoproline-containing Acm halves, PPL 0 and PPL 2, in mycelial suspensions of Streptomyces antibioticus...
January 12, 2018: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/29327656/nad-nadh-homeostasis-affects-metabolic-adaptation-to-hypoxia-and-secondary-metabolite-production-in-filamentous-fungi
#8
Motoyuki Shimizu
Filamentous fungi are used to produce fermented foods, organic acids, beneficial secondary metabolites and various enzymes. During such processes, these fungi balance cellular NAD+:NADH ratios to adapt to environmental redox stimuli. Cellular NAD(H) status in fungal cells is a trigger of changes in metabolic pathways including those of glycolysis, fermentation, and the production of organic acids, amino acids and secondary metabolites. Under hypoxic conditions, high NADH:NAD+ ratios lead to the inactivation of various dehydrogenases, and the metabolic flow involving NAD+ is down-regulated compared with normoxic conditions...
January 12, 2018: Bioscience, Biotechnology, and Biochemistry
https://www.readbyqxmd.com/read/29327513/methanol-water-aqueous-phase-reforming-by-the-assistant-of-dehydrogenases-at-near-room-temperature
#9
Yangbin Shen, Yulu Zhan, Shuping Li, Fandi Ning, Ying Du, Yunjie Huang, Ting He, Xiaochun Zhou
As an excellent hydrogen storage medium, methanol has many advantages, such as high hydrogen content (12.6 wt%), low-cost and availability from biomass or photocatalysis. However, conventional methanol-water reforming usually proceeds at high temperature. In this research, we successfully designed a new effective strategy to generate hydrogen from methanol at near-room temperature. The strategy involves two main procedures, which are CH3OH→HCOOH→H2 and NADH→HCOOH→H2. The first procedure (CH3OH→HCOOH→H2) is performed by alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH) and the Ir-catalyst...
January 11, 2018: ChemSusChem
https://www.readbyqxmd.com/read/29324667/dietary-resveratrol-does-not-affect-life-span-body-composition-stress-response-and-longevity-related-gene-expression-in-drosophila-melanogaster
#10
Stefanie Staats, Anika E Wagner, Bianca Kowalewski, Florian T Rieck, Sebastian T Soukup, Sabine E Kulling, Gerald Rimbach
In this study, we tested the effect of the stilbene resveratrol on life span, body composition, locomotor activity, stress response, and the expression of genes encoding proteins centrally involved in ageing pathways in the model organism Drosophila melanogaster. Male and female w1118 D. melanogaster were fed diets based on sucrose, corn meal, and yeast. Flies either received a control diet or a diet supplemented with 500 µmol/L resveratrol. Dietary resveratrol did not affect mean, median, and maximal life span of male and female flies...
January 11, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29323490/activity-related-microsecond-dynamics-revealed-by-temperature-jump-f%C3%A3-rster-resonance-energy-transfer-measurements-on-thermophilic-alcohol-dehydrogenase
#11
Morgan B Vaughn, Jianyu Zhang, Thomas G Spiro, R Brian Dyer, Judith P Klinman
Previous studies of a thermophilic alcohol dehydrogenase (ht-ADH) demonstrated a range of discontinuous transitions at 30 °C that include catalysis, kinetic isotope effects, protein hydrogen-deuterium exchange rates, and intrinsic fluorescence properties. Using the Förster resonance energy transfer response from a Trp-NADH donor-acceptor pair in T-jump studies of ht-ADH, we now report microsecond protein motions that can be directly related to active site chemistry. Two distinctive transients are observed: a slow, kinetic process lacking a temperature break, together with a faster transient that is only detectable above 30 °C...
January 11, 2018: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/29322853/laser-ablation-of-the-biliary-tree-in-vivo-proof-of-concept-as-potential-treatment-of-unresectable-cholangiocarcinoma
#12
Paola Saccomandi, Giuseppe Quero, Riccardo Gassino, Alfonso Lapergola, Ludovica Guerriero, Michele Diana, Alberto Vallan, Guido Perrone, Emiliano Schena, Guido Costamagna, Jaques Marescaux, Francesco M Di Matteo
OBJECTIVES: The palliative treatment of cholangiocarcinoma is based on stent placement with well-known procedure-related complications. Consequently, alternative energy-based techniques were put forward with controversial long-term results. This study aims to evaluate the safety and effectiveness of biliary tree laser ablation (LA) in terms of: i) absence of perforation, ii) temperature increase, iii) induced thermal damage in in vivo models. MATERIALS AND METHODS: The common bile duct and cystic ducts of two pigs were ablated with a diode laser (circumferential irradiation pattern) for 6 and 3 minutes at 7 W...
January 11, 2018: International Journal of Hyperthermia
https://www.readbyqxmd.com/read/29322283/rational-design-of-engineered-microbial-cell-surface-multi-enzyme-co-display-system-for-sustainable-nadh-regeneration-from-low-cost-biomass
#13
Lei Han, Bo Liang, Jianxia Song
As an important cofactor, NADH is essential for most redox reactions and biofuel cells. However, supply of exogenous NADH is challenged, due to the low production efficiency and high cost of NADH regeneration system, as well as low stability of NADH. Here, we constructed a novel cell surface multi-enzyme co-display system with ratio- and space-controllable manner as exogenous NADH regeneration system for the sustainable NADH production from low-cost biomass. Dockerin-fused glucoamylase (GA) and glucose dehydrogenase (GDH) were expressed and assembled on the engineered bacterial surfaces, which displayed protein scaffolds with various combinations of different cohesins...
January 10, 2018: Journal of Industrial Microbiology & Biotechnology
https://www.readbyqxmd.com/read/29321463/use-of-a-smartphone-as-a-colorimetric-analyzer-in-paper-based-devices-for-sensitive-and-selective-determination-of-mercury-in-water-samples
#14
Purim Jarujamrus, Rattapol Meelapsom, Somkid Pencharee, Apinya Obma, Maliwan Amatatongchai, Nadh Ditcharoen, Sanoe Chairam, Suparb Tamuang
A smartphone application, called CAnal, was developed as a colorimetric analyzer in paper-based devices for sensitive and selective determination of mercury(II) in water samples. Measurement on the double layer of a microfluidic paper-based analytical device (μPAD) fabricated by alkyl ketene dimer (AKD)-inkjet printing technique with special design doped with unmodified silver nanoparticles (AgNPs) onto the detection zones was performed by monitoring the gray intensity in the blue channel of AgNPs, which disintegrated when exposed to mercury(II) on μPAD...
2018: Analytical Sciences: the International Journal of the Japan Society for Analytical Chemistry
https://www.readbyqxmd.com/read/29317887/genotyping-echinococcus-granulosus-from-canine-isolates-in-ilam-province-west-of-iran
#15
Abdolhossein Dalimi, Morteza Shamsi, Afra Khosravi, Fatemeh Ghaffarifar
Background: Cystic Echinococcosis (CE) is one of the most common parasitic zoonosis caused by Echinococcus granulosus worldwide. This study investigated genotype diversity of Echinococcus granulosus isolated from stray dogs and golden jackals in Ilam province, West of Iran. Methods: Adult worms were collected from the small intestine of the stray dogs and golden jackals from Ilam Province roads during 2012-2014. DNA was extracted from the adult worms and the partial mitochondrial NADH dehydrogenase subunit1 (nad1) was amplified by PCR then the products were digested by using HpaII, Rsa1, and Alu1 restriction enzymes...
October 2017: Iranian Journal of Parasitology
https://www.readbyqxmd.com/read/29317025/electrochemical-sensing-platform-based-on-kelp-derived-hierarchical-meso-macroporous-carbons
#16
Yashuang Hei, Xiqian Li, Xiao Zhou, Jingju Liu, Mimi Sun, Tianze Sha, Cuixing Xu, Wanyi Xue, Xiangjie Bo, Ming Zhou
In this paper, kelp (Laminaria japonica), as a kind of abundant biomass, is used as the precursor for the preparation of kelp-derived hierarchical meso-macroporous carbons (K-dHMMCs) through the carbonization under nitrogen (N2) atmosphere at high temperature. The K-dHMMCs exhibits the unique structure with high specific surface area of 416.02 m2 g-1, large pore volume of 0.24 cm3 g-1, the hierarchical meso-macroporous size distribution centered at 2, 12 and 82 nm and high density of defective sites, enabling K-dHMMCs attractive for the electrocatalysis...
March 20, 2018: Analytica Chimica Acta
https://www.readbyqxmd.com/read/29316934/physiological-mitochondrial-and-oxidative-stress-differences-in-the-presence-or-absence-of-lactation-in-rats
#17
Hayden W Hyatt, Yufeng Zhang, Wendy R Hood, Andreas N Kavazis
BACKGROUND: Human epidemiological data show that breastfeeding reduces the mother's probability of developing several disease conditions, including obesity and type II diabetes compared to mothers that give birth but do not breastfeed. The goal of this investigation was to characterize how lactation changes a rat's body composition, metabolism, mitochondrial function, and oxidative stress. METHODS: Ten-week old female Sprague-Dawley rats were divided into three groups (n = 8 per group): 1) non-reproductive (NR), 2) those that were allowed to mate and give birth, but were not allowed to suckle their pups (PP), and 3) those that were allowed to mate and give birth, and suckled their young until weaning at 21 days (PL)...
January 9, 2018: Reproductive Biology and Endocrinology: RB&E
https://www.readbyqxmd.com/read/29311464/-development-and-application-of-catalytic-tyrosine-modification
#18
Shinichi Sato, Michihiko Tsushima, Kosuke Nakamura, Hiroyuki Nakamura
 The chemical labeling of proteins with synthetic probes is a key technique used in chemical biology, protein-based therapy, and material science. Much of the chemical labeling of native proteins, however, depends on the labeling of lysine and cysteine residues. While those methods have significantly contributed to native protein labeling, alternative methods that can modify different amino acid residues are still required. Herein we report the development of a novel methodology of tyrosine labeling, inspired by the luminol chemiluminescence reaction...
2018: Yakugaku Zasshi: Journal of the Pharmaceutical Society of Japan
https://www.readbyqxmd.com/read/29311229/integrated-analysis-of-proteomic-and-transcriptomic-data-highlights-late-fetal-muscle-maturation-process
#19
Valentin Voillet, Magali San Cristobal, Marie-Christine Pere, Yvon Billon, Laurianne Canario, Laurence Liaubet, Louis Lefaucheur
BACKGROUND: In pigs, the perinatal period is the most critical time for survival. Piglet maturation, which occurs at the end of gestation, leads to a state of full development after birth. Maturity is thus an important determinant of early survival. Skeletal muscle plays a key role in adaptation to extra-uterine life, e.g. motor function and thermoregulation. Progeny from two breeds with extreme neonatal mortality rates were analyzed at 90 and 110 days of gestation (dg). The Large White breed is a highly selected breed for lean growth with a high rate of mortality at birth, whereas the Chinese Meishan breed is fatter and more robust and has a low mortality rate...
January 8, 2018: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/29303599/proteomic-analysis-of-peroxynitrite-induced-protein-nitration-in-isolated-beef-heart-mitochondria
#20
M Kohutiar, A Eckhardt, I Mikšík, P Šantorová, J Wilhelm
Mitochondria are exposed to reactive nitrogen species under physiological conditions and even more under several pathologic states. In order to reveal the mechanism of these processes we studied the effects of peroxynitrite on isolated beef heart mitochondria in vitro. Peroxynitrite has the potential to nitrate protein tyrosine moieties, break the peptide bond, and eventually release the membrane proteins into the solution. All these effects were found in our experiments. Mitochondrial proteins were resolved by 2D electrophoresis and the protein nitration was detected by immunochemical methods and by nano LC-MS/MS...
January 5, 2018: Physiological Research
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