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https://www.readbyqxmd.com/read/28641210/comparative-analysis-of-polyspecificity-of-the-endogenous-trna-synthetase-of-different-expression-host-towards-photocrosslinking-amino-acids-using-an-in-silico-approach
#1
Nadarajan Saravanan Prabhu, Hyungdon Yun
Photo-induced covalent crosslinking has emerged as the powerful strategy for analyzing and characterizing the protein-protein interaction and mapping protein 3D conformations. In the last decades, a number of photocrosslinking amino acids have been reported but only a few have been efficiently utilized for photocrosslinking purposes. Recently, incorporation of diazirine containing photoactivatable analogs such as photo-methionine, photo-leucine, photo-isoleucine and photo-lysine into target proteins were accomplished in live cells (Human A549cells, HEK 293) by depleting corresponding natural amino acid and supplementing these analogs in the medium...
June 8, 2017: Journal of Molecular Graphics & Modelling
https://www.readbyqxmd.com/read/28636132/trpc1-and-trpc3-dependent-ca-2-signaling-in-mouse-cortical-astrocytes-affects-injury-evoked-astrogliosis-in-vivo
#2
Thabet Belkacemi, Alexander Niermann, Laura Hofmann, Ulrich Wissenbach, Lutz Birnbaumer, Petra Leidinger, Christina Backes, Eckart Meese, Andreas Keller, Xianshu Bai, Anja Scheller, Frank Kirchhoff, Stephan E Philipp, Petra Weissgerber, Veit Flockerzi, Andreas Beck
Following brain injury astrocytes change into a reactive state, proliferate and grow into the site of lesion, a process called astrogliosis, initiated and regulated by changes in cytoplasmic Ca(2+) . Transient receptor potential canonical (TRPC) channels may contribute to Ca(2+) influx but their presence and possible function in astrocytes is not known. By RT-PCR and RNA sequencing we identified transcripts of Trpc1, Trpc2, Trpc3, and Trpc4 in FACS-sorted glutamate aspartate transporter (GLAST)-positive cultured mouse cortical astrocytes and subcloned full-length Trpc1 and Trpc3 cDNAs from these cells...
June 21, 2017: Glia
https://www.readbyqxmd.com/read/28632911/polymer-functionalized-gold-nanoparticles-as-non-viral-gene-delivery-reagents
#3
M Mar Encabo-Berzosa, Maria Sancho-Albero, Victor Sebastian, Silvia Irusta, Manuel Arruebo, Jesus Santamaria, Pilar Martín Duque
BACKGROUND: In this study we investigated the ability of PEG functionalized gold nanoparticles as non-viral vectors in the transfection of different cell lines, comparing them with commercial lipoplexes. METHODS: Positively charged gold nanoparticles were synthesized using PEI as reducing and stabilizer agent and its cytotoxicity reduced by its functionalization with PEG. We bound the nanoparticles to three plasmids with different sizes (4-40 kpb). The vector internalization was evaluated by confocal and electronic microscopy...
June 20, 2017: Journal of Gene Medicine
https://www.readbyqxmd.com/read/28631939/a-high-throughput-screening-assay-for-nkcc1-cotransporter-using-nonradioactive-rubidium-flux-technology
#4
Sikander Gill, Rajwant Gill, Yang Wen, Thilo Enderle, Doris Roth, Dong Liang
A high-throughput screening (HTS) assay was developed for cotransporter, NKCC1, which is a potential target for the treatment of diverse disorders. This nonradioactive rubidium flux assay coupled with ion channel reader series provides a working screen for this target expressed in human embryonic kidney (HEK) cell line. An eightfold window of detection was achieved with the optimized assay. This new functional assay offered a robust working model for NKCC1 in determining reliable and concordant rank orders of the test compounds supporting its sensitivity and specificity...
May 2017: Assay and Drug Development Technologies
https://www.readbyqxmd.com/read/28630066/porphyromonas-gulae-activates-unprimed-and-ifn%C3%AE-primed-macrophages-via-the-pattern-recognition-receptors-tlr-2-tlr-4-and-nod2
#5
James A Holden, Neil M O'Brien-Simpson, Jason C Lenzo, Rebecca K H Orth, Ashley Mansell, Eric C Reynolds
Porphyromonas gulae is an anaerobic, Gram-negative coccobacillus that has been associated with periodontal disease in companion animals. The aims of this study were to analyse the ligation of pattern recognition receptors by P. gulae and the subsequent activation of macrophages. Exposure of HEK cells transfected with Toll-like receptors or NOD-like receptors to P. gulae resulted in the ligation of TLR2, TLR4 and NOD2. The effect of this engagement of receptors was investigated by measuring the synthesis of nitric oxide, CD86 expression and inflammatory cytokine production using wild type, TLR2-/- and TLR4-/- macrophages...
June 19, 2017: Infection and Immunity
https://www.readbyqxmd.com/read/28624448/identification-of-functional-hypoxia-inducible-factor-response-elements-in-the-human-lysyl-oxidase-gene-promoter
#6
Victoria Wang, David A Davis, Robert Yarchoan
Human lysyl oxidase (LOX) is a hypoxia-responsive gene whose product catalyzes collagen crosslinking and is thought to be important in cancer metastasis and osteoarthritis. We previously demonstrated that LOX was upregulated by hypoxia inducible factor 2 (HIF-2) more strongly than hypoxia inducible 1 (HIF-1). We further investigated the response of the LOX gene and LOX promoter to HIFs. LOX mRNA, measured by real time reverse transcriptase-PCR, was strongly up-regulated (almost 40-fold), by transfection of HEK-293T cells with a plasmid encoding the HIF-2α subunit of HIF-2, but only three-fold by a plasmid encoding HIF-1α...
June 14, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28622852/efficacy-of-e2-glycoprotein-fused-to-porcine-cd154-as-a-novel-chimeric-subunit-vaccine-to-prevent-classical-swine-fever-virus-vertical-transmission-in-pregnant-sows
#7
Sara Muñoz-González, Yusmel Sordo, Marta Pérez-Simó, Marisela Suárez, Albert Canturri, Maria Pilar Rodriguez, María Teresa Frías-Lepoureau, Mariano Domingo, Mario Pablo Estrada, Llilianne Ganges
Here we evaluated the effect of double vaccination with a novel subunit marker vaccine candidate based in the CSFV E2 glycoprotein fused to the porcine CD154 to prevent CSFV vertical transmission. A lentivirus-based gene delivery system was used to obtain a stable recombinant HEK 293 cell line for the expression of E2 fused to porcine CD154 molecule. Six pregnant sows were distributed in two groups and at 64days of gestation animals numbered 1-4 (group 1) were vaccinated via intramuscular inoculation with 50μg of E2-CD154 subunit vaccine...
June 2017: Veterinary Microbiology
https://www.readbyqxmd.com/read/28620305/comparative-effects-of-chloride-channel-inhibitors-on-lrrc8-vrac-mediated-chloride-conductance
#8
Jonas Friard, Michel Tauc, Marc Cougnon, Vincent Compan, Christophe Duranton, Isabelle Rubera
Chloride channels play an essential role in a variety of physiological functions and in human diseases. Historically, the field of chloride channels has long been neglected owing to the lack of powerful selective pharmacological agents that are needed to overcome the technical challenge of characterizing the molecular identities of these channels. Recently, members of the LRRC8 family have been shown to be essential for generating the volume-regulated anion channel (VRAC) current, a chloride conductance that governs the regulatory volume decrease (RVD) process...
2017: Frontiers in Pharmacology
https://www.readbyqxmd.com/read/28618136/amphiphilic-cargo-loaded-nanocarrier-enhances-antibiotic-uptake-and-perturbs-efflux-effective-synergy-for-mitigation-of-methicillin-resistant-staphylococcus-aureus
#9
Durairaj Thiyagarajan, Gopal Das, Ramesh Aiyagari
A pyridinium amphiphile-loaded Poly(lactic-co-glycolic acid) (PLGA) nanocarrier (C1-PNC) was developed as an adjuvant in order to break the resistance and restore susceptibility of methicillin-resistant Staphylococcus aureus (MRSA) cells to therapeutic antibiotics. Notably, against a clinical MRSA strain, C1-PNC could render 8 × and 6 × reduction of the minimum biofilm eradication concentration (MBEC90) of gentamicin and ciprofloxacin, respectively. Mechanistic studies on MRSA planktonic cells revealed that in case of gentamicin, C1-PNC promoted enhanced cellular uptake of the antibiotic, while the propensity of C1-PNC to inhibit efflux pump activity could be leveraged to enhance cellular accumulation of ciprofloxacin leading to effective killing of MRSA cells...
June 15, 2017: ChemMedChem
https://www.readbyqxmd.com/read/28614363/rigid-geometry-solves-curse-of-dimensionality-effects-in-clustering-methods-an-application-to-omics-data
#10
Shun Adachi
The quality of samples preserved long term at ultralow temperatures has not been adequately studied. To improve our understanding, we need a strategy to analyze protein degradation and metabolism at subfreezing temperatures. To do this, we obtained liquid chromatography-mass spectrometry (LC/MS) data of calculated protein signal intensities in HEK-293 cells. Our first attempt at directly clustering the values failed, most likely due to the so-called "curse of dimensionality". The clusters were not reproducible, and the outputs differed with different methods...
2017: PloS One
https://www.readbyqxmd.com/read/28613134/elevation-of-mir-9-3p-suppresses-the-epithelial-mesenchymal-transition-of-nasopharyngeal-carcinoma-cells-via-down-regulating-fn1-itgb1-and-itgav
#11
Yu Ding, Yinghua Pan, Shan Liu, Feng Jiang, Junbo Jiao
MicroRNAs had been proved to be pivotal regulators in nasopharyngeal carcinoma (NPC) by regulating a large amount of genes' expression. In our research, we aim to explore the functions of miR-9-3p on the metastases of NPC and figure out the potential mechanisms. First, we revealed downregulation of miR-9-3p and upregulation of fibronectin 1 (FN1), β1 integrin (ITGB1) and α5 integrin (ITGAV) expression in NPC tissues and cells compared with the normal using RNA-seq analysis, RT-qPCR, western blot and immunohistochemistry...
June 14, 2017: Cancer Biology & Therapy
https://www.readbyqxmd.com/read/28611375/identification-of-an-in-vivo-orally-active-dual-binding-protein-protein-interaction-inhibitor-targeting-tnf%C3%AE-through-combined-in-silico-in-vitro-in-vivo-screening
#12
Hadley Mouhsine, Hélène Guillemain, Gabriel Moreau, Najla Fourati, Chouki Zerrouki, Bruno Baron, Lucille Desallais, Patrick Gizzi, Nesrine Ben Nasr, Julie Perrier, Rojo Ratsimandresy, Jean-Louis Spadoni, Hervé Do, Patrick England, Matthieu Montes, Jean-François Zagury
TNFα is a homotrimeric pro-inflammatory cytokine, whose direct targeting by protein biotherapies has been an undeniable success for the treatment of chronic inflammatory diseases. Despite many efforts, no orally active drug targeting TNFα has been identified so far. In the present work, we identified through combined in silico/in vitro/in vivo approaches a TNFα direct inhibitor, compound 1, displaying nanomolar and micromolar range bindings to TNFα. Compound 1 inhibits the binding of TNFα with both its receptors TNFRI and TNFRII...
June 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28608407/disulfide-bonds-and-disorder-in-granulin-3-an-unusual-handshake-between-structural-stability-and-plasticity
#13
Gaurav Ghag, Christopher J Holler, Georgia Taylor, Thomas L Kukar, Vladimir N Uversky, Vijayaraghavan Rangachari
Granulins (GRNs) are a family of small (∼6 kDa) proteins generated by the proteolytic processing of their precursor, progranulin (PGRN), in many cell types. Both PGRN and GRNs are implicated in a plethora of biological functions, often in opposing roles to each other. Lately, GRNs have generated significant attention due to their implicated roles in neurodegenerative disorders. Despite their physiological and pathological significance, the structure-function relationships of GRNs are poorly defined. GRNs contain 12 conserved cysteines forming six intramolecular disulfide bonds, making them rather exceptional, even among a few proteins with high disulfide bond density...
June 12, 2017: Protein Science: a Publication of the Protein Society
https://www.readbyqxmd.com/read/28605224/effect-of-a-cooling-kit-on-physiology-and-performance-following-exercise-in-the-heat
#14
Cody R Smith, Cory L Butts, J D Adams, Matthew A Tucker, Nicole E Moyen, Matthew S Ganio, Brendon P McDermott
CONTEXT: Exercising in the heat leads to an increase in body temperature that can increase the risk of heat illness or cause detriments in exercise performance. OBJECTIVE: To examine a phase change heat emergency kit (HEK) on thermoregulatory and perceptual responses, and subsequent exercise performance following exercise in the heat. DESIGN: Two randomized crossover trials which consisted of 30 minutes of exercise, 15 minutes of treatment (T1), performance testing (5-10-5 pro-agility test, 1500 m run), and another 15 minutes of treatment (T2) identical to T1...
June 12, 2017: Journal of Sport Rehabilitation
https://www.readbyqxmd.com/read/28604766/the-deubiquitinating-enzymes-usp4-and-usp17-target-hyaluronan-synthase-2-and-differentially-affect-its-function
#15
M Mehić, V K de Sa, S Hebestreit, C-H Heldin, P Heldin
The levels of hyaluronan, a ubiquitous glycosaminoglycan prominent in the extracellular matrix, is balanced through the actions of hyaluronan-synthesizing enzymes (HAS1, 2 and 3) and degrading hyaluronidases (Hyal 1, 2, 3 and PH20). Hyaluronan accumulates in rapidly remodeling tissues, such as breast cancer, due to deregulated expression of the HAS2 gene and/or alterations of HAS2 activity. The activity of HAS2 is regulated by post-translational modifications, including ubiquitination. In order to identify deubiquitinating enzymes (DUBs) that are involved in de-ubiquitination of HAS2, a complementary (cDNA) library of 69 Flag-HA-tagged human DUBs cloned into retroviral vectors was screened in human embryonic kidney (HEK) 293T cells for their ability to de-ubiquitinate myc-tagged HAS2...
June 12, 2017: Oncogenesis
https://www.readbyqxmd.com/read/28603497/sigma-1-receptor-plays-a-negative-modulation-on-n-type-calcium-channel
#16
Kang Zhang, Zhe Zhao, Liting Lan, Xiaoli Wei, Liyun Wang, Xiaoyan Liu, Haitao Yan, Jianquan Zheng
The sigma-1 receptor is a 223 amino acids molecular chaperone with a single transmembrane domain. It is resident to eukaryotic mitochondrial-associated endoplasmic reticulum and plasma membranes. By chaperone-mediated interactions with ion channels, G-protein coupled receptors and cell-signaling molecules, the sigma-1 receptor performs broad physiological and pharmacological functions. Despite sigma-1 receptors have been confirmed to regulate various types of ion channels, the relationship between the sigma-1 receptor and N-type Ca(2+) channel is still unclear...
2017: Frontiers in Pharmacology
https://www.readbyqxmd.com/read/28603085/kaempferol-increases-levels-of-coenzyme-q-in-kidney-cells-and-serves-as-a-biosynthetic-ring-precursor
#17
Lucía Fernández-Del-Río, Anish Nag, Elena Gutiérrez Casado, Julia Ariza, Agape M Awad, Akil I Joseph, Ohyun Kwon, Eric Verdin, Rafael de Cabo, Claus Schneider, Jorge Z Torres, María I Burón, Catherine F Clarke, José M Villalba
Coenzyme Q (Q) is a lipid-soluble antioxidant essential in cellular physiology. Patients with Q deficiencies, with few exceptions, seldom respond to treatment. Current therapies rely on dietary supplementation with Q10, but due to its highly lipophilic nature, Q10 is difficult to absorb by tissues and cells. Plant polyphenols, present in the human diet, are redox active and modulate numerous cellular pathways. In the present study, we tested whether treatment with polyphenols affected the content or biosynthesis of Q...
June 9, 2017: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/28599308/an-ultra-sensitive-biophysical-risk-assessment-of-light-effect-on-skin-cells
#18
Devasier Bennet, Buddolla Viswanath, Sanghyo Kim, Jeong Ho An
The aim of this study was to analyze photo-dynamic and photo-pathology changes of different color light radiations on human adult skin cells. We used a real-time biophysical and biomechanics monitoring system for light-induced cellular changes in an in vitro model to find mechanisms of the initial and continuous degenerative process. Cells were exposed to intermittent, mild and intense (1-180 min) light with On/Off cycles, using blue, green, red and white light. Cellular ultra-structural changes, damages, and ECM impair function were evaluated by up/down-regulation of biophysical, biomechanical and biochemical properties...
May 24, 2017: Oncotarget
https://www.readbyqxmd.com/read/28598898/a-99mtc-labelled-scfv-antibody-fragment-that-binds-to-prostate-specific-membrane-antigen
#19
Saima Nawaz, Gregory E D Mullen, Philip J Blower, James R Ballinger
INTRODUCTION: Prostate-specific membrane antigen (PSMA) is an extensively studied antigen for imaging prostate cancer. We prepared a single-chain variable fragment (scFv) of J591, a monoclonal antibody that recognises an external epitope of PSMA, incorporating a His-tag for labelling with Tc tricarbonyl, and evaluated its binding using human PCa cell lines. METHODS: J591(scFv) was expressed in HEK-293T cells and purified by metal ion affinity chromatography, followed by size exclusion chromatography...
June 8, 2017: Nuclear Medicine Communications
https://www.readbyqxmd.com/read/28597972/a-novel-interacting-protein-serp1-regulates-the-n-linked-glycosylation-and-function-of-glp-1-receptor-in-the-liver
#20
Yuanyuan Xiao, Junfeng Han, Qianqian Wang, Yueqin Mao, Meilin Wei, Weiping Jia, Li Wei
Glucagon-like peptide 1 (GLP-1) exerts multiple effects on metabolism through its receptor, GLP-1R, in the liver. Activation and transduction of GLP-1R require complex interactions of largely unknown accessory proteins, and these processes are crucial to the response to endoplasmic reticulum (ER) stress. Using the membrane-based split ubiquitin yeast two-hybrid system (MYTH) and a human liver cDNA library, we obtained the human GLP-1R interactome and identified SERP1 as a potential interacting protein based on its ability to stabilize membrane proteins and facilitate N-linked glycosylation...
June 9, 2017: Journal of Cellular Biochemistry
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