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https://www.readbyqxmd.com/read/29224228/newcastle-disease-virus-ndv-co-expressing-il7-and-il15-modified-tumor-cells-as-a-vaccine-for-cancer-immunotherapy
#1
Xiaojing Xu, Qing Sun, Yu Mei, Yonghao Liu, Lixiang Zhao
IL15 and IL7 are two cytokines essential for T cell development and homeostasis. In order to improve the antitumor activity by Newcastle Disease Virus (NDV)-modified tumor vaccine, we generated a recombinant NDV co-expressing IL15 and IL7 (LX/IL(15+7)) by incorporation of a 2A self-processing peptide into IL15 and IL7 using reverse genetics. B16 cells infected with LX/IL(15+7) expressed both IL15 and IL7 stably. The cytotoxicity assay showed that murine melanoma cells modified with LX/IL(15+7) could significantly enhance the antitumor immune response in vitro...
December 10, 2017: Cancer Science
https://www.readbyqxmd.com/read/29224168/tet-assisted-bisulfite-sequencing-tab-seq
#2
Miao Yu, Dali Han, Gary C Hon, Chuan He
5-Hydroxymethylcytosine (5hmC) is a modified form of cytosine, which has recently been found in mammalian cells and tissues. 5hmC is derived from 5-methylcytosine (5mC) by Ten-eleven translocation (TET) family protein-mediated oxidation and may regulate gene expression. Numerous affinity-based profiling methods have been developed to help understand the exact function of 5hmC in the genome. However, these methods have a relatively low resolution (~100 bp) without quantitative information of the modification percentage on each site...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29224128/identification-of-a-new-begomovirus-infecting-duranta-repens-in-pakistan
#3
Sidra Anwar, Muhammad Tahir
Ornamental perennial plants may serve as reservoirs for viruses that infect field crops. Duranta repens is an ornamental shrub that frequently exhibits leaf curl symptoms typical of begomoviral infection. Two begomoviruses or DNA-A components (isolates 57SA and 58SA) along with a DNA-B component (31SA) were identified in a symptomatic D. repens plant. Isolates 57SA and 58SA showed the highest nucleotide sequence identity of 90% and 92.2% to catharanthus yellow mosaic virus (CaYMV) and chilli leaf curl India virus (ChiLCINV), respectively, whereas 31SA shared the highest nucleotide sequence identity of 95...
December 9, 2017: Archives of Virology
https://www.readbyqxmd.com/read/29224069/clickseq-replacing-fragmentation-and-enzymatic-ligation-with-click-chemistry-to-prevent-sequence-chimeras
#4
Elizabeth Jaworski, Andrew Routh
We recently reported a fragmentation-free method for the synthesis of Next-Generation Sequencing libraries called "ClickSeq" that uses biorthogonal click-chemistry in place of enzymes for the ligation of sequencing adaptors. We found that this approach dramatically reduces artifactual chimera formation, allowing the study of rare recombination events that include viral replication intermediates and defective-interfering viral RNAs. ClickSeq illustrates how robust, bio-orthogonal chemistry can be harnessed in vitro to capture and dissect complex biological processes...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29223735/targeting-secreted-cytokine-bmp9-gates-the-attenuation-of-hepatic-fibrosis
#5
Peng Li, Yongyun Li, Liqi Zhu, Zhi Yang, Jie He, Lihua Wang, Qingfeng Shang, Hui Pan, Huixue Wang, Xiong Ma, Bin Li, Xianqun Fan, Shengfang Ge, Renbing Jia, He Zhang
Liver fibrosis is overly exuberant wound healing that leads to portal hypertension or liver cirrhosis. Recent studies have demonstrated the functions of bone morphogenetic protein 9 (BMP9) in liver fibrosis, and thus, targeting liver-specific BMP9 abnormalities will become an attractive approach for developing therapeutics to treat liver fibrosis. Here, we reveal that BMP9 serves as a valuable serum diagnostic indicator and efficient therapeutic target to attenuate liver fibrogenesis. Our analysis of biopsies from liver fibrotic patients revealed that higher BMP9 levels accompanied advanced stages of liver fibrosis...
December 6, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/29223485/preventive-effect-of-anti-vaca-egg-yolk-immunoglobulin-igy-on-helicobacter-pylori-infected-mice
#6
Kyung Sook Hong, Mi-Ran Ki, H M Arif Ullah, Eun-Joo Lee, Yong Deuk Kim, Myung-Jin Chung, Ahmed K Elfadl, Jin-Kyu Park, Kyu-Shik Jeong
BACKGROUND: Helicobacter pylori, a gram-negative bacterium, is the causative agent of gastric disorders and gastric cancer in the human stomach. Vacuolating cytotoxin A (VacA) is among the multi-effect protein toxins released by H. pylori that enables its persistence in the human stomach. METHODS: To evaluate the effect of anti-VacA egg yolk immunoglobulin (anti-VacA IgY) on H. pylori infection, a highly specific anti-VacA IgY was produced from egg yolks of hens immunized with a mixture of two purified recombinant VacAs...
December 6, 2017: Vaccine
https://www.readbyqxmd.com/read/29223463/terfenadine-metabolism-of-human-cytochrome-p450-2j2-containing-genetic-variations-g312r-p351l-and-p115l
#7
Dabin Jeong, Hyoung-Goo Park, Young-Ran Lim, Yejin Lee, Vitchan Kim, Myung-A Cho, Donghak Kim
The human cytochrome P450 2J2 is involved in several metabolic reactions, including the oxidation of important therapeutics and epoxidation of endogenous arachidonic acid. At least ten genetic variations of P450 2J2 have been identified, but their effects on enzymatic activity have not been clearly characterized. Here, we evaluated the functional effects of three genetic variations of P450 2J2 (G312R, P351L, and P115L). Recombinant enzymes of wild-type and three variant P450 2J2 were heterologously expressed in Escherichia coli and purified...
November 11, 2017: Drug Metabolism and Pharmacokinetics
https://www.readbyqxmd.com/read/29223397/-borneol-suppresses-conditioned-fear-recall-and-anxiety-like-behaviors-in-mice
#8
Bo Cao, Huan-Yu Ni, Jun Li, Ying Zhou, Xin-Lan Bian, Yan Tao, Cheng-Yun Cai, Cheng Qin, Hai-Yin Wu, Lei Chang, Chun-Xia Luo, Dong-Ya Zhu
Fear- and anxiety-related psychiatric disorders have been one of the major chronic diseases afflicting patients for decades, and new compounds for treating such disorders remain to be developed. (+)-Borneol, a bicyclic monoterpene found in several species of Artemisia and Dipterocarpaceae, is widely used for anxiety, pain and anesthesia in Chinese medicine. Meanwhile, it can potentiate GABA (γ-aminobutyric acid) activity directly in recombinant GABAA receptors. The present study was to investigate the effects of (+)-Borneol on both contextual and cued fear recall...
December 6, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29223348/distinctive-binding-properties-of-the-negative-allosteric-modulator-3h-sb269-652-at-recombinant-dopamine-d3-receptors
#9
Irene Fasciani, Ilaria Pietrantoni, Mario Rossi, Clotilde Mannoury la Cour, Gabriella Aloisi, Francesco Marampon, Marco Scarselli, Mark J Millan, Roberto Maggio
Recently, employing radioligand displacement and functional coupling studies, we demonstrated that SB269,652 (N-[(1r,4r)-4-[2-(7-cyano-1,2,3,4-tetrahydroisoquinolin-2-yl)ethyl]cyclohexyl]-1H-indole-2-carboxamide) interacts in an atypical manner with dopamine D3 receptor displaying a unique profile reminiscent of a negative allosteric ligand. Here, we characterized the binding of radiolabelled [3H]SB269,652 to human dopamine D3 receptor stably expressed in Chinese Hamster Ovary cells. Under saturating conditions, SB269,652 showed a KD value of ≈ 1nM...
December 6, 2017: European Journal of Pharmacology
https://www.readbyqxmd.com/read/29223338/molecular-cloning-and-expression-analysis-of-aquaporin-genes-in-pearl-millet-pennisetum-glaucum-l-r-br-genotypes-contrasting-in-their-transpiration-response-to-high-vapour-pressure-deficits
#10
Palakolanu Sudhakar Reddy, Murugesan Tharanya, Kaliamoorthy Sivasakthi, Mallayee Srikanth, C Tom Hash, Jana Kholova, Kiran K Sharma, Vincent Vadez
Pearl millet is a crop of the semi-arid tropics having high degree of genetic diversity and variable tolerance to drought stress. To investigate drought tolerance mechanism that possibly accounts for differences in drought tolerance, four recombinant inbred lines from a high resolution cross (HRC) were selected for variability in their transpiration rate (Tr) response to vapour pressure deficit (VPD) conditions. The differential Tr response of the genotypes to increased VPD conditions was used to classify the genotypes as sensitive or insensitive to high VPD...
December 2017: Plant Science: An International Journal of Experimental Plant Biology
https://www.readbyqxmd.com/read/29223162/spectral-kinetic-analysis-of-recombination-reaction-of-heme-centers-of-bd-type-quinol-oxidase-from-escherichia-coli-with-carbon-monoxide
#11
S A Siletsky, A V Dyuba, D A Elkina, M V Monakhova, V B Borisov
Recombination of the isolated, fully reduced bd-type quinol oxidase from Escherichia coli with carbon monoxide was studied by pulsed absorption spectrophotometry with microsecond time resolution. Analysis of the kinetic phases of recombination was carried out using the global analysis of multiwavelength kinetic data ("Global fitting"). It was found that the unresolved photodissociation of CO is followed by a stepwise (with four phases) recombination with characteristic times (τ) of about 20 µs, 250 µs, 1...
November 2017: Biochemistry. Biokhimii︠a︡
https://www.readbyqxmd.com/read/29223159/factors-beyond-enolase-2-and-mitochondrial-lysyl-trna-synthetase-precursor-are-required-for-trna-import-into-yeast-mitochondria
#12
M V Baleva, M Meyer, N Entelis, I Tarassov, P Kamenski, B Masquida
In yeast, the import of tRNALys with CUU anticodon (tRK1) relies on a complex mechanism where interaction with enolase 2 (Eno2p) dictates a deep conformational change of the tRNA. This event is believed to mask the tRNA from the cytosolic translational machinery to re-direct it towards the mitochondria. Once near the mitochondrial outer membrane, the precursor of the mitochondrial lysyl-tRNA synthetase (preMsk1p) takes over enolase to carry the tRNA within the mitochondrial matrix, where it is supposed to participate in translation following correct refolding...
November 2017: Biochemistry. Biokhimii︠a︡
https://www.readbyqxmd.com/read/29223157/self-organization-of-recombinant-membrane-porin-ompf-from-yersinia-pseudotuberculosis-in-aqueous-environments
#13
E V Sidorin, V A Khomenko, N Yu Kim, P S Dmitrenok, A M Stenkova, O D Novikova, T F Solov'eva
Recombinant porin OmpF (an integral protein of bacterial outer membrane) from Yersinia pseudotuberculosis was synthesized in Escherichia coli cells as inclusion bodies. By combining the methods of anion-exchange and gel filtration chromatographies, recombinant OmpF (rOmpF) was isolated as an individual protein in its denatured state, and its characteristic properties (molecular mass, N-terminal amino acid sequence, and hydrodynamic radius of the protein in 8 M urea solution) were determined. According to the data of gel filtration, dynamic light scattering, optical spectroscopy, and binding of the hydrophobic fluorescent probe 8-anilino-1-naphthalenesulfonic acid, the rOmpF is fully unfolded in 8 M urea and exists in random coil conformation...
November 2017: Biochemistry. Biokhimii︠a︡
https://www.readbyqxmd.com/read/29223156/biochemical-properties-and-phylogeny-of-hydroxypyruvate-reductases-from-methanotrophic-bacteria-with-different-c1-assimilation-pathways
#14
S Y But, S V Egorova, V N Khmelenina, Y A Trotsenko
In the aerobic methanotrophic bacteria Methylomicrobium alcaliphilum 20Z, Methylococcus capsulatus Bath, and Methylosinus trichosporium OB3b, the biochemical properties of hydroxypyruvate reductase (Hpr), an indicator enzyme of the serine pathway for assimilation of reduced C1-compounds, were comparatively analyzed. The recombinant Hpr obtained by cloning and heterologous expression of the hpr gene in Escherichia coli catalyzed NAD(P)H-dependent reduction of hydroxypyruvate or glyoxylate, but did not catalyze the reverse reactions of D-glycerate or glycolate oxidation...
November 2017: Biochemistry. Biokhimii︠a︡
https://www.readbyqxmd.com/read/29223155/recombinant-human-erythropoietin-with-additional-processable-protein-domains-purification-of-protein-synthesized-in-escherichia-coli-heterologous-expression-system
#15
T M Grunina, A V Demidenko, A M Lyaschuk, M S Poponova, Z M Galushkina, L A Soboleva, S A Cherepushkin, N B Polyakov, D A Grumov, A I Solovyev, V G Zhukhovitsky, I S Boksha, M E Subbotina, A V Gromov, V G Lunin, A S Karyagina
Three variants of human recombinant erythropoietin (rhEPO) with additional N-terminal protein domains were obtained by synthesis in an Escherichia coli heterologous expression system. These domains included (i) maltose-binding protein (MBP), (ii) MBP with six histidine residues (6His) in N-terminal position, (iii) s-tag (15-a.a. oligopeptide derived from bovine pancreatic ribonuclease A) with N-terminal 6His. Both variants of the chimeric protein containing MBP domain were prone to aggregation under nondenaturing conditions, and further purification of EPO after the domain cleavage by enterokinase proved to be impossible...
November 2017: Biochemistry. Biokhimii︠a︡
https://www.readbyqxmd.com/read/29223152/electron-transfer-through-the-acceptor-side-of-photosystem-i-interaction-with-exogenous-acceptors-and-molecular-oxygen
#16
REVIEW
D A Cherepanov, G E Milanovsky, A A Petrova, A N Tikhonov, A Yu Semenov
This review considers the state-of-the-art on mechanisms and alternative pathways of electron transfer in photosynthetic electron transport chains of chloroplasts and cyanobacteria. The mechanisms of electron transport control between photosystems (PS) I and II and the Calvin-Benson cycle are considered. The redistribution of electron fluxes between the noncyclic, cyclic, and pseudocyclic pathways plays an important role in the regulation of photosynthesis. Mathematical modeling of light-induced electron transport processes is considered...
November 2017: Biochemistry. Biokhimii︠a︡
https://www.readbyqxmd.com/read/29223136/allele-exchange-at-the-epsps-locus-confers-glyphosate-tolerance-in-cassava
#17
Aaron W Hummel, Raj Deepika Chauhan, Tomas Cermak, Andrew M Mutka, Anupama Vijayaraghavan, Adam Boyher, Colby G Starker, Rebecca Bart, Daniel F Voytas, Nigel J Taylor
Effective weed control can protect yields of cassava (Manihot esculenta) storage roots (Doll and Piedrahita Cañola, 1978). Farmers could benefit from using herbicide with a tolerant cultivar. We applied traditional transgenesis and gene editing to generate robust glyphosate tolerance in cassava. By comparing promoters regulating expression of transformed 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) genes with various paired amino acid substitutions, we found that strong constitutive expression is required to achieve glyphosate tolerance during in vitro selection and in whole cassava plants...
December 9, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/29223031/protonography-and-anion-inhibition-profile-of-the-%C3%AE-carbonic-anhydrase-cruca4-identified-in-the-mediterranean-red-coral-corallium-rubrum
#18
Sonia Del Prete, Daniela Vullo, Natacha Caminiti-Segonds, Didier Zoccola, Sylvie Tambutté, Claudiu T Supuran, Clemente Capasso
CruCA4 is a secreted isoform of the α-carbonic anhydrase (CA, EC 4.2.1.1) family, which has been identified in the octocoral Corallium rubrum. This enzyme is involved in the calcification process leading to the formation of the coral calcium carbonate skeleton. We report here experiments performed on the recombinant CruCA4 with the technique of protonography that can be used to detect in a simple way the enzyme activity. We have also investigated the inhibition profile of CruCA4 with one major class of CA inhibitors, the inorganic anions...
December 5, 2017: Bioorganic Chemistry
https://www.readbyqxmd.com/read/29223005/the-emerging-role-of-systems-biology-for-engineering-protein-production-in-cho-cells
#19
REVIEW
Chih-Chung Kuo, Austin Wt Chiang, Isaac Shamie, Mojtaba Samoudi, Jahir M Gutierrez, Nathan E Lewis
To meet the ever-growing demand for effective, safe, and affordable protein therapeutics, decades of intense efforts have aimed to maximize the quantity and quality of recombinant proteins produced in CHO cells. Bioprocessing innovations and cell engineering efforts have improved product titer; however, uncharacterized cellular processes and gene regulatory mechanisms still hinder cell growth, specific productivity, and protein quality. Herein, we summarize recent advances in systems biology and data-driven approaches aiming to unravel how molecular pathways, cellular processes, and extrinsic factors (e...
December 6, 2017: Current Opinion in Biotechnology
https://www.readbyqxmd.com/read/29222842/kinetic-characterisation-of-wild-type-and-mutant-human-thioredoxin-glutathione-reductase-defines-its-reaction-and-regulatory-mechanisms
#20
Christina Brandstaedter, Karin Fritz-Wolf, Stine Weder, Marina Fischer, Beate Hecker, Stefan Rahlfs, Katja Becker
In most cells the thioredoxin (Trx) and glutathione systems are essentially involved in maintaining redox homeostasis. The selenoprotein thioredoxin glutathione reductase (TGR) is a hybrid enzyme in which a glutaredoxin (Grx) domain is linked to a thioredoxin reductase (TrxR). Notably, the protein is also capable of reducing glutathione disulfide (GSSG) thus representing an important link between the two redox systems. In this study, we recombinantly produced human TGR (hTGR wild type) by fusing its open reading frame with a bacterial SECIS element and co-expressing the construct in E...
December 8, 2017: FEBS Journal
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