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https://www.readbyqxmd.com/read/28199384/basic-studies-on-epigenetic-carcinogenesis-of-low-dose-exposure-to-1-trichloromethyl-1-2-3-4-tetrahydro-%C3%AE-carboline-taclo-in-vitro
#1
Renjie Wang, Yi Cui, Yi Xu, Joseph Irudayaraj
1-Trichloromethyl-1,2,3,4-tetrahydro-β-carboline (TaClo) has been widely studied as a neurotoxic substance, however, only few reports have explored its effect on carcinogenicity. Since the aberrant modification of DNA methylation occurs very early in almost all human cancers, the focus of this study is to assess the carcinogenicity of TaClo by characterizing alterations of the epigenetic state, specifically, DNA methylation, upon exposure to TaClo in a HEK 293 model cell line. Our results suggest that TaClo could induce global DNA hypomethylation and transcriptional repression of critical tumor suppressor genes by increasing their promoter methylation...
2017: PloS One
https://www.readbyqxmd.com/read/28198671/transcriptome-analysis-reveals-dynamic-changes-in-coxsackievirus-a16-infected-hek-293t-cells
#2
Jun Jin, Rujiao Li, Chunlai Jiang, Ruosi Zhang, Xiaomeng Ge, Fang Liang, Xin Sheng, Wenwen Dai, Meili Chen, Jiayan Wu, Jingfa Xiao, Weiheng Su
BACKGROUND: Coxsackievirus A16 (CVA16) and enterovirus 71 (EV71) are two of the major causes of hand, foot and mouth disease (HFMD) world-wide. Although many studies have focused on infection and pathogenic mechanisms, the transcriptome profile of the host cell upon CVA16 infection is still largely unknown. RESULTS: In this study, we compared the mRNA and miRNA expression profiles of human embryonic kidney 293T cells infected and non-infected with CVA16. We highlighted that the transcription of SCARB2, a cellular receptor for both CVA16 and EV71, was up-regulated by nearly 10-fold in infected cells compared to non-infected cells...
January 25, 2017: BMC Genomics
https://www.readbyqxmd.com/read/28198589/synthesis-and-characterization-of-stimuli-responsive-star-like-polypept-o-ides-introducing-biodegradable-peptostars
#3
Regina Holm, Benjamin Weber, Philipp Heller, Kristina Klinker, Dana Westmeier, Dominic Docter, Roland H Stauber, Matthias Barz
Star-like polymers are one of the smallest systems in the class of core crosslinked polymeric nanoparticles. This article reports on a versatile, straightforward synthesis of three-arm star-like polypept(o)ide (polysarcosine-block-polylysine) polymers, which are designed to be either stable or degradable at elevated levels of glutathione. Polypept(o)ides are a recently introduced class of polymers combining the stealth-like properties of the polypeptoid polysarcosine with the functionality of polypeptides, thus enabling the synthesis of materials completely based on endogenous amino acids...
February 15, 2017: Macromolecular Bioscience
https://www.readbyqxmd.com/read/28197552/altered-channel-conductance-states-and-gating-of-gabaa-receptors-by-a-pore-mutation-linked-to-dravet-syndrome
#4
Ciria C Hernandez, Weijing Kong, Ningning Hu, Yujia Zhang, Wangzhen Shen, Laurel Jackson, Xiaoyan Liu, Yuwu Jiang, Robert L Macdonald
We identified a de novo missense mutation, P302L, in the γ-aminobutyric acid type A (GABAA) receptor γ2 subunit gene GABRG2 in a patient with Dravet syndrome using targeted next-generation sequencing. The mutation was in the cytoplasmic portion of the transmembrane segment M2 of the γ2 subunit that faces the pore lumen. GABAA receptor α1 and β3 subunits were coexpressed with wild-type (wt) γ2L or mutant γ2L(P302L) subunits in HEK 293T cells and cultured mouse cortical neurons. We measured currents using whole-cell and single-channel patch clamp techniques, surface and total expression levels using surface biotinylation and Western blotting, and potential structural perturbations in mutant GABAA receptors using structural modeling...
January 2017: ENeuro
https://www.readbyqxmd.com/read/28193781/real-time-imaging-of-sodium-glucose-transporter-sglt1-trafficking-and-activity-in-single-cells
#5
Chiara Ghezzi, Guillaume Calmettes, Pauline Morand, Bernard Ribalet, Scott John
The processes controlling targeting of glucose transporters to apical and basolateral membranes of polarized cells are complex and not-well understood. We have engineered SGLT1 and GLUT4 constructs linked to fluorescent proteins to highlight the differences in transporter expression and trafficking, in real time, in different cell types. Activity was assessed in parallel using a FRET glucose sensor. In COS cells and HEK cells, SGLT1 was distributed between the plasma membrane and intracellular compartments, but there was little expression in CHO cells...
February 2017: Physiological Reports
https://www.readbyqxmd.com/read/28188528/lumier-a-discovery-tool-for-mammalian-protein-interaction-networks
#6
Miriam Barrios-Rodiles, Jonathan D Ellis, Benjamin J Blencowe, Jeffrey L Wrana
Protein-protein interactions (PPIs) play an essential role in all biological processes. In vivo, PPIs occur dynamically and depend on extracellular cues. To discover novel protein-protein interactions in mammalian cells, we developed a high-throughput automated technology called LUMIER (LUminescence-based Mammalian IntERactome). In this approach, we co-express a Luciferase (LUC)-tagged fusion protein along with a Flag-tagged protein in an efficiently transfectable cell line such as HEK-293T cells. The interaction between the two proteins is determined by co-immunoprecipitation using an anti-Flag antibody, and the presence of the LUC-tagged interactor in the complex is subsequently detected via its luciferase activity...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28182310/ankyrin-rich-membrane-spanning-protein-as-a-novel-modulator-of-transient-receptor-potential-vanilloid-1-function-in-nociceptive-neurons
#7
J Peter, C Kasper, M Kaufholz, R Buschow, J Isensee, T Hucho, F W Herberg, F Schwede, C Stein, S-E Jordt, M Brackmann, V Spahn
BACKGROUND: The ion channel TRPV1 is mainly expressed in small diameter dorsal root ganglion (DRG) neurons, which are involved in the sensation of acute noxious thermal and chemical stimuli. Direct modifications of the channel by diverse signalling events have been intensively investigated, but little is known about the composition of modulating macromolecular TRPV1 signalling complexes. Here, we hypothesize that the novel adaptor protein ankyrin-rich membrane spanning protein/kinase D interacting substrate (ARMS) interacts with TRPV1 and modulates its function in rodent DRG neurons...
February 9, 2017: European Journal of Pain: EJP
https://www.readbyqxmd.com/read/28181895/-inter-practice-variation-in-polypharmacy-prevalence-amongst-older-patients-in-primary-care
#8
J Sinnige, J C Braspenning, F G Schellevis, K Hek, I Stirbu, G P Westert, J C Korevaar
PURPOSE: Complex medication management in older people with multiple chronic conditions can introduce practice variation in polypharmacy prevalence. This study aimed to determine the inter-practice variation in polypharmacy prevalence and examine how this variation was influenced by patient and practice characteristics. METHODS: This cohort study included 45,731 patients aged 55 years and older with at least one prescribed medication from 126 general practices that participated in NIVEL Primary Care Database in the Netherlands...
2017: Nederlands Tijdschrift Voor Geneeskunde
https://www.readbyqxmd.com/read/28168212/novel-synaptobrevin-1-mutation-causes-fatal-congenital-myasthenic-syndrome
#9
Xin-Ming Shen, Rosana H Scola, Paulo J Lorenzoni, Cláudia S K Kay, Lineu C Werneck, Joan Brengman, Duygu Selcen, Andrew G Engel
OBJECTIVE: To identify the molecular basis and elucidate the pathogenesis of a fatal congenital myasthenic syndrome. METHODS: We performed clinical electrophysiology studies, exome and Sanger sequencing, and analyzed functional consequences of the identified mutation. RESULTS: Clinical electrophysiology studies of the patient revealed several-fold potentiation of the evoked muscle action potential by high frequency nerve stimulation pointing to a presynaptic defect...
February 2017: Annals of Clinical and Translational Neurology
https://www.readbyqxmd.com/read/28165042/relationship-between-auxiliary-gamma-subunits-and-mallotoxin-on-bk-channel-modulation
#10
Xin Guan, Qin Li, Jiusheng Yan
The large-conductance, calcium- and voltage-activated K(+)(BK) channel consists of the pore-forming α subunits (BKα) and auxiliary subunits. The auxiliary γ1-3 subunits potently modulate the BK channel by shifting its voltage-dependence of channel activation toward the hyperpolarizing direction by approximately 145 mV (γ1), 100 mV (γ2), and 50 mV (γ3). Mallotoxin is a potent small-molecule BK channel activator. We analyzed the relationship between mallotoxin and the γ subunits in their BK channel-activating effects in membrane patches excised from HEK-293 cells...
February 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28163207/expression-of-recombinant-surfactant-protein-sfta3-in-the-human-kidney-cell-line-hek-293t
#11
H Schröder, R Fischer, L Sollfrank, F Paulsen, L Bräuer, M Schicht
Pulmonary surfactant is broadly known to keep the lung dry, clean and open by lowering the surface tension of the fluid-film that lines the alveoli. The surfactant's protein component, the so called surfactant proteins (SPs), make up a multifunctional protein family. In addition to the four "classical" surfactant proteins (SP-A, SP-B, SP-C and SP-D), which possess immunologic as well as surfactant regulatory properties, two novel putative surfactant proteins (SFTA2 and SFTA3) have recently been described. Neither of them shows sequential nor structural similarity with the already known surfactant proteins...
February 3, 2017: Annals of Anatomy, Anatomischer Anzeiger: Official Organ of the Anatomische Gesellschaft
https://www.readbyqxmd.com/read/28161964/in-vitro-evaluation-of-a-polytope-dna-construct-as-a-novel-dna-vaccine-strategy-against-human-cytomegalovirus-associated-diseases
#12
E Sabbaghian, F Roodbari, J Amani, A Rafiei
Human cytomegalovirus (HCMV) establishes severe disease in fetus, newborn and immunocompromised individuals. Polytope DNA vaccine strategy allows us to choose conserved and immunodominant epitopes from different antigens that can stimulate cellular and humoral immune responses simultaneously. In this study, a synthetic chimeric gene fragment was subcloned in to DNA vaccine vector pcDNA3.1+. The recombinant vector was transferred in to suitable eukaryotic cell line HEK 293T and the expression level of polytope construct from HEK 293T-infected cells was determined by western blot...
February 6, 2017: Acta Virologica
https://www.readbyqxmd.com/read/28160563/tumor-suppressor-role-of-mir-3622b-5p-in-erbb2-positive-cancer
#13
Mingjie Lu, Tongshan Wang, Mingfeng He, Wenfang Cheng, Ting Yan, Zebo Huang, Lan Zhang, Huo Zhang, Wei Zhu, Yichao Zhu, Ping Liu
Over-expression or amplification of ERBB2 is observed in multifarious carcinomas. However, the molecular mechanism of ERBB2 downregulation in ERBB2-positive cancers remains obscure. This experiment investigated the suppressive role of miR-3622b-5p in ERBB2-positive breast and gastric cancers. The luciferase activity of ERBB2 3'-untranslated region-based reporters constructed in HEK-293T, SK-BR-3 and MCF-10A cells suggested that ERBB2 was the target gene of miR-3622b-5p. Over-expressed miR-3622b-5p reduced the protein level of ERBB2, weakened the activation of mTORC1/S6, and induced the apoptosis of ERBB2-positive cancer cells...
February 1, 2017: Oncotarget
https://www.readbyqxmd.com/read/28159417/design-synthesis-and-cytotoxicity-of-stabilized-mycolactone-analogs
#14
Vaddela Sudheer Babu, Ya Zhou, Yoshito Kishi
On exposure to visible light, mycolactone A/B, the causative toxin of Buruli ulcer, rearranges to a mixture of four photo-mycolactones apparently via a rare photochemically-induced [4πs+2πa] cycloaddition. In order to prevent the rearrangement, two C6'-C7' dihydromycolactone analogs 6'α-15 and 6'β-15 were designed and synthesized. 6'α-15 and 6'β-15 were shown to be stable under not only photochemical, but also acidic and basic conditions. Cytotoxicity was tested against arbitrarily chosen four cell lines (human Hek-293, human lung carcinoma A-549, human melanoma LOX-IMVI, and mouse L-929), thereby revealing that: (1) both analogs maintain potent cytotoxicity; (2) 6'β-15 exhibits significantly higher potency against human cell lines than 6'α-15; (3) in comparison with parent mycolactone A/B, 6'β-15 exhibits equal potency against human Hek-293, whereas significantly lower potency against human lung carcinoma A-549 and human melanoma LOX-IMVI...
January 17, 2017: Bioorganic & Medicinal Chemistry Letters
https://www.readbyqxmd.com/read/28154813/the-effect-of-selective-d-or-n-%C3%AE-methyl-arginine-substitution-on-the-activity-of-the-proline-rich-antimicrobial-peptide-chex1-arg20
#15
Wenyi Li, Zhe Sun, Neil M O'Brien-Simpson, Laszlo Otvos, Eric C Reynolds, Mohammed A Hossain, Frances Separovic, John D Wade
In vivo pharmacokinetics studies have shown that the proline-rich antimicrobial peptide, A3-APO, which is a discontinuous dimer of the peptide, Chex1-Arg20, undergoes degradation to small fragments at positions Pro6-Arg7 and Val19-Arg20. With the aim of minimizing or abolishing this degradation, a series of Chex1-Arg20 analogs were prepared via Fmoc/tBu solid phase peptide synthesis with D-arginine or, in some cases, peptide backbone N(α)-methylated arginine, substitution at these sites. All the peptides were tested for antibacterial activity against the Gram-negative bacterium Klebsiella pneumoniae...
2017: Frontiers in Chemistry
https://www.readbyqxmd.com/read/28152550/novel-11%C3%AE-hydroxysteroid-dehydrogenase-1-inhibitors-reduce-cortisol-levels-in-keratinocytes-and-improve-dermal-collagen-content-in-human-ex-vivo-skin-after-exposure-to-cortisone-and-uv
#16
Stéphanie M Boudon, Anna Vuorinen, Piero Geotti-Bianchini, Eliane Wandeler, Denise V Kratschmar, Marc Heidl, Remo Campiche, Eileen Jackson, Alex Odermatt
Activity and selectivity assessment of new bi-aryl amide 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1) inhibitors, prepared in a modular manner via Suzuki cross-coupling, are described. Several compounds inhibiting 11β-HSD1 at nanomolar concentrations were identified. Compounds 2b, 3e, 7b and 12e were shown to selectively inhibit 11β-HSD1 over 11β-HSD2, 17β-HSD1 and 17β-HSD2. These inhibitors also potently inhibited 11β-HSD1 activity in intact HEK-293 cells expressing the recombinant enzyme and in intact primary human keratinocytes expressing endogenous 11β-HSD1...
2017: PloS One
https://www.readbyqxmd.com/read/28150939/antinociceptive-effects-of-a-novel-%C3%AE-2-%C3%AE-3-subtype-selective-gabaa-receptor-positive-allosteric-modulator
#17
Lakeisha A Lewter, Janet L Fisher, Justin N Siemian, Kashi Reddy Methuku, Michael M Poe, James M Cook, Jun-Xu Li
Pain remains a challenging clinical condition and spinal GABAA receptors are crucial modulators of pain processing. α2/α3-subtype GABAA receptors mediate the analgesic actions of benzodiazepines. Positive allosteric modulators (PAMs) at α2/α3-subtype GABAA receptors may have analgesic potential. Here we report a new selective α2/α3-subtype GABAA receptor PAM in in vitro and in vivo pain assays. KRM-II-81 demonstrated similar efficacy at α1/α2/α3 GABAA receptors and negligible efficacy at α4/α5/α6 GABAA receptors, with α2 and α3-subtypes being 17- and 28-fold more potent than α1 subtypes in HEK-293T cells expressing GABAA receptors with different α subunits...
February 2, 2017: ACS Chemical Neuroscience
https://www.readbyqxmd.com/read/28145731/coupling-of-airway-smooth-muscle-tas2rs-to-intracellular-signaling-and-relaxation-is-via-g%C3%AE-i1-2-3
#18
Donghwa Kim, Jung A Woo, Ezekiel Geffken, Steven S An, Stephen B Liggett
Bitter taste receptors (TAS2Rs) are expressed on human airway smooth muscle (HASM) and evoke marked relaxation. Agonist interaction with TAS2Rs activates PLC and increases compartmentalized [Ca(2+)]i via IP3. In taste cells, the G-protein gustducin couples TAS2R to PLC, however we find very low levels of Gαgust mRNA or protein in HASM. We hypothesized that another G-protein in HASM transmits TAS2R function. TAS2R signaling to [Ca(2+)]i, ERK1/2, and physiologic relaxation was sensitive to pertussis toxin confirming a role for a member of the Gi-family...
February 1, 2017: American Journal of Respiratory Cell and Molecular Biology
https://www.readbyqxmd.com/read/28137449/the-binding-orientations-of-structurally-related-ligands-can-differ-a-cautionary-note
#19
Marc-David Ruepp, Hao Wei, Michele Leuenberger, Martin Lochner, Andrew J Thompson
Crystal structures can identify ligand-receptor interactions and assist the development of novel therapeutics, but experimental challenges sometimes necessitate the use of homologous proteins. Tropisetron is an orthosteric ligand at both 5-HT3 and α7 nACh receptors and its binding orientation has been determined in the structural homologue AChBP (pdbid: 2WNC). Co-crystallisation with a structurally-related ligand, granisetron, reveals an almost identical orientation (pdbid; 2YME). However, there is a >1000-fold difference in the affinity of tropisetron at 5-HT3 versus α7 nACh receptors, and α7 nACh receptors do not bind granisetron...
January 27, 2017: Neuropharmacology
https://www.readbyqxmd.com/read/28132817/systematic-mapping-of-rna-chromatin-interactions-in%C3%A2-vivo
#20
Bharat Sridhar, Marcelo Rivas-Astroza, Tri C Nguyen, Weizhong Chen, Zhangming Yan, Xiaoyi Cao, Lucie Hebert, Sheng Zhong
RNA molecules can attach to chromatin. It remains difficult to know what RNAs are associated with chromatin and where the genomic target loci of these RNAs are. Here, we present MARGI (mapping RNA-genome interactions), a technology to massively reveal native RNA-chromatin interactions from unperturbed cells. The gist of this technology is to ligate chromatin-associated RNAs (caRNAs) with their target genomic sequences by proximity ligation, forming RNA-DNA chimeric sequences, which are converted to a sequencing library for paired-end sequencing...
January 25, 2017: Current Biology: CB
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