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https://www.readbyqxmd.com/read/28437636/chimeric-flock-house-virus-protein-a-with-endoplasmic-reticulum-targeting-domain-enhances-viral-replication-and-virus-like-particle-trans-encapsidation-in-plants
#1
Yiyang Zhou, Christopher M Kearney
Flock House virus (FHV) RNA can be trans-encapsidated, entirely in planta, by tobacco mosaic virus coat protein to form virus-like particles (VLPs). Vaccination with these VLPs leads to strong antigen expression in mice and immune-activation. We hypothesize that creating an additional cellular site for replication and/or trans-encapsidation might significantly improve the final output of trans-encapsidated product. FHV protein A was engineered to target the endoplasmic reticulum (ER) via a heterologous tobacco etch virus ER-targeting domain, and was expressed in cis or in trans relative to the replicating FHV RNA1...
April 21, 2017: Virology
https://www.readbyqxmd.com/read/28437617/electrostatic-origin-of-red-solvatochromic-shift-of-dfhbdi-in-rna-spinach
#2
Samik Bose, Suman Chakrabarty, Debashree Ghosh
Interactions with environment tune the spectral properties of biological chromophores, e.g. fluorescent proteins. Understanding of the relative contribution of the various types of non-covalent interactions in the spectral shifts can provide rational design principles towards developing new fluorescent probes. In this work, we investigate the origin of the red-shift in the absorption spectra of difluoro hydroxybenzylidene dimethyl imidazolinone (DFHBDI) chromophore in RNA Spinach as compared to the aqueous solution...
April 24, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/28437464/inhibition-of-%C3%AE-v%C3%AE-3-integrin-induces-loss-of-cell-directionality-of-oral-squamous-carcinoma-cells-oscc
#3
Cyntia F Montenegro, Bruna C Casali, Rafael L B Lino, Bianca C Pachane, Patty K Santos, Alan R Horwitz, Heloisa S Selistre-de-Araujo, Marcelo L Lamers
The connective tissue formed by extracellular matrix (ECM) rich in fibronectin and collagen consists a barrier that cancer cells have to overpass to reach blood vessels and then a metastatic site. Cell adhesion to fibronectin is mediated by αvβ3 and α5β1 integrins through an RGD motif present in this ECM protein, thus making these receptors key targets for cell migration studies. Here we investigated the effect of an RGD disintegrin, DisBa-01, on the migration of human fibroblasts (BJ) and oral squamous cancer cells (OSCC, SCC25) on a fibronectin-rich environment...
2017: PloS One
https://www.readbyqxmd.com/read/28437166/virus-like-particles-of-mrna-with-artificial-minimal-coat-proteins-particle-formation-stability-and-transfection-efficiency
#4
Shehrazade Jekhmane, Rob de Haas, Omar Paulino da Silva Filho, Alexander H van Asbeck, Marco Emanuele Favretto, Armando Hernandez Garcia, Roland Brock, Renko de Vries
RNA has enormous potential as a therapeutic, yet, the successful application depends on efficient delivery strategies. In this study, we demonstrate that a designed artificial viral coat protein, which self-assembles with DNA to form rod-shaped virus-like particles (VLPs), also encapsulates and protects mRNA encoding enhanced green fluorescent protein (EGFP) and luciferase, and yields cellular expression of these proteins. The artificial viral coat protein consists of an oligolysine (K12) for binding to the oligonucleotide, a silk protein-like midblock S10 = (GAGAGAGQ)10 that self-assembles into stiff rods, and a long hydrophilic random coil block C that shields the nucleic acid cargo from its environment...
February 21, 2017: Nucleic Acid Therapeutics
https://www.readbyqxmd.com/read/28437137/differential-requirement-of-the-ribosomal-protein-s6-and-ribosomal-protein-s6-kinase-for-plant-virus-accumulation-and-interaction-of-s6-kinase-with-potyviral-vpg
#5
Minna-Liisa Rajamäki, Dehui Xi, Sidona Sikorskaite-Gudziuniene, Jari P T Valkonen, Steven A Whitham
Ribosomal protein S6 (RPS6) is an indispensable plant protein regulated, in part, by ribosomal protein S6 kinase (S6K) which, in turn, is a key regulator of plant responses to stresses and developmental cues. Increased expression of RPS6 was detected in Nicotiana benthamiana during infection by diverse plant viruses. Silencing of the RPS6 and S6K genes in N. benthamiana affected accumulation of Cucumber mosaic virus, Turnip mosaic virus (TuMV), and Potato virus A (PVA) in contrast to Turnip crinkle virus and Tobacco mosaic virus...
April 24, 2017: Molecular Plant-microbe Interactions: MPMI
https://www.readbyqxmd.com/read/28436958/local-clearance-of-senescent-cells-attenuates-the-development-of-post-traumatic-osteoarthritis-and-creates-a-pro-regenerative-environment
#6
Ok Hee Jeon, Chaekyu Kim, Remi-Martin Laberge, Marco Demaria, Sona Rathod, Alain P Vasserot, Jae Wook Chung, Do Hun Kim, Yan Poon, Nathaniel David, Darren J Baker, Jan M van Deursen, Judith Campisi, Jennifer H Elisseeff
Senescent cells (SnCs) accumulate in many vertebrate tissues with age and contribute to age-related pathologies, presumably through their secretion of factors contributing to the senescence-associated secretory phenotype (SASP). Removal of SnCs delays several pathologies and increases healthy lifespan. Aging and trauma are risk factors for the development of osteoarthritis (OA), a chronic disease characterized by degeneration of articular cartilage leading to pain and physical disability. Senescent chondrocytes are found in cartilage tissue isolated from patients undergoing joint replacement surgery, yet their role in disease pathogenesis is unknown...
April 24, 2017: Nature Medicine
https://www.readbyqxmd.com/read/28436953/disruption-of-direct-3d-telomere-trf2-interaction-through-two-molecularly-disparate-mechanisms-is-a-hallmark-of-primary-hodgkin-and-reed-sternberg-cells
#7
Hans Knecht, Nathalie A Johnson, Tina Haliotis, Daniel Lichtensztejn, Sabine Mai
In classical Hodgkin's lymphoma (cHL), specific changes in the 3D telomere organization cause progression from mononuclear Hodgkin cells (H) to multinucleated Reed-Sternberg cells (RS). In a post-germinal center B-cell in vitro model, permanent latent membrane protein 1 (LMP1) expression, as observed in Epstein-Barr virus (EBV)-associated cHL, results in multinuclearity and complex chromosomal aberrations through downregulation of key element of the shelterin complex, the telomere repeat binding factor 2 (TRF2)...
April 24, 2017: Laboratory Investigation; a Journal of Technical Methods and Pathology
https://www.readbyqxmd.com/read/28436559/dopamine-d1-or-d2-receptor-expressing-neurons-in-the-central-nervous-system
#8
Xiaoyan Wei, Tengfei Ma, Yifeng Cheng, Cathy C Y Huang, Xuehua Wang, Jiayi Lu, Jun Wang
Dopamine signals mainly through D1 receptors (D1Rs) and D2 receptors (D2Rs); D1R-expressing or D2R-expressing neurons contribute to distinct reward and addictive behaviors. Traditionally, transgenic mice expressing green fluorescent protein (GFP) under D1R or D2R promoters are used for fluorescent verification in electrophysiology studies, whereas Cre mice are employed for behavioral research. However, it is unknown whether the same neuronal populations are targeted in GFP and Cre mice. Additionally, while D1Rs and D2Rs are known to be expressed in different striatal neurons, their expression patterns outside the striatum remain unclear...
April 24, 2017: Addiction Biology
https://www.readbyqxmd.com/read/28436474/interactions-of-amtn-odam-and-scpppq1-proteins-of-a-specialized-basal-lamina-that-attaches-epithelial-cells-to-tooth-mineral
#9
Aurélien Fouillen, Juliana Dos Santos Neves, Charline Mary, Jean-Daniel Castonguay, Pierre Moffatt, Christian Baron, Antonio Nanci
A specialized basal lamina (sBL) mediates adhesion of certain epithelial cells to the tooth. It is distinct because it does not contain collagens type IV and VII, is enriched in laminin-332, and includes three novel constituents called amelotin (AMTN), odontogenic ameloblast-associated (ODAM), and secretory calcium-binding phosphoprotein proline-glutamine rich 1 (SCPPPQ1). The objective of this study was to clarify the structural organization of the sBL. Fluorescence and immunogold labeling showed that the three proteins co-localize...
April 24, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28436469/a-novel-method-to-evaluate-ribosomal-performance-in-cell-free-protein-synthesis-systems
#10
Noémie Kempf, Cristina Remes, Ralph Ledesch, Tina Züchner, Henning Höfig, Ilona Ritter, Alexandros Katranidis, Jörg Fitter
Cell-free protein synthesis (CFPS) systems were designed to produce proteins with a minimal set of purified components, thus offering the possibility to follow translation as well as protein folding. In order to characterize the performance of the ribosomes in such a system, it is crucial to separately quantify the two main components of productivity, namely the fraction of active ribosomes and the number of synthesizing cycles. Here, we provide a direct and highly reliable measure of ribosomal activity in any given CFPS system, introducing an enhanced-arrest peptide variant...
April 24, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28436453/functional-thermodynamics-structural-and-biological-studies-of-in-silico-identified-inhibitors-of-mycobacterium-tuberculosis-enoyl-acp-coa-reductase-enzyme
#11
Leonardo K B Martinelli, Mariane Rotta, Anne D Villela, Valnês S Rodrigues-Junior, Bruno L Abbadi, Rogério V Trindade, Guilherme O Petersen, Giuliano M Danesi, Laura R Nery, Ivani Pauli, Maria M Campos, Carla D Bonan, Osmar Norberto de Souza, Luiz A Basso, Diogenes S Santos
Novel chemotherapeutics agents are needed to kill Mycobacterium tuberculosis, the main causative agent of tuberculosis (TB). The M. tuberculosis 2-trans-enoyl-ACP(CoA) reductase enzyme (MtInhA) is the druggable bona fide target of isoniazid. New chemotypes were previously identified by two in silico approaches as potential ligands to MtInhA. The inhibition mode was determined by steady-state kinetics for seven compounds that inhibited MtInhA activity. Dissociation constant values at different temperatures were determined by protein fluorescence spectroscopy...
April 24, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28436335/localization-of-the-werner-protein-together-with-h2ax-in-%C3%AE-irradiation-induced-neoplastic-transformed-human-mesenchymal-stem-cells
#12
Nedime Serakinci, Tufan Cankaya, Steen Kølvraa
The H2A histone family, member X (H2AX), and Werner (WRN) are important proteins for genome and telomere maintenance. WRN has a major role in genome stability, particularly during DNA replication, transcription, recombination, and repair of DNA double-stranded breaks (DSBs) via base excision repair, homologous recombination, or nonhomologous end joining. H2AX plays a part in the rapid, sensitive, cellular response to the ionizing radiation or DNA-damaging chemotherapeutic agents that cause DSBs. This occurs when radiation-induced DSBs trigger the activation of H2AX and begin the damage-repair process...
2017: Critical Reviews in Eukaryotic Gene Expression
https://www.readbyqxmd.com/read/28436225/watsonianone-a-from-rhodomyrtus-tomentosa-fruit-attenuates-respiratory-syncytial-virus-induced-inflammation-in-vitro
#13
Ling Zhuang, Li-Feng Chen, Yu-Bo Zhang, Zhong Liu, Xu-Hui Xiao, Wei Tang, Guo-Cai Wang, Wen-Jun Song, Yao-Lan Li, Man-Mei Li
Respiratory syncytial virus (RSV) is one of the most common respiratory pathogens. Immoderate inflammation plays a great role in causing RSV-induced diseases. In the present study, watsonianone A, isolated from the fruit of Rhodomyrtus tomentosa (Ait.) Hassk, was found to show a good inhibitory effect on RSV-induced NO production, with a half-maximal inhibitory concentration of 37.2 ± 1.6 μM. Enzyme-linked immunosorbent assay and fluorescence quantitative polymerase chain reaction analyses indicated that watsonianone A markedly reduced both mRNA and protein levels of tumor necrosis factor α, interleukin 6, and monocyte chemoattractant protein 1 in RSV-infected RAW264...
April 24, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28436037/high-level-ab-initio-absorption-spectra-simulations-of-neutral-anionic-and-neutral-chromophore-of-green-fluorescence-protein-chromophore-models-in-gas-phase-and-solution
#14
Ivelina Georgieva, Adelia J A Aquino, Natasha Trendafilova, Hans Lischka
Semi-classical ab initio simulations of the absorption spectra of neutral and anionic p-hydroxybenzylidene-2,3-dimethylimidazolinone (p-HBDI), a model chromophore of Green Fluorescent Protein (GFP) and of a positively charged neutral (N+)-HBDI chromophore model were performed in gas phase with the resolution-of-identity algebraic diagrammatic construction through second-order (RI-ADC(2)) method. The calculated absorption spectra in gas phase are composed of one band centered at 3.51 eV (HBDI), 2.50 eV (HBDI(-) ) and 3...
April 24, 2017: Photochemistry and Photobiology
https://www.readbyqxmd.com/read/28436014/a-critical-assessment-of-the-synthesis-and-biological-activity-of-p53-hdm2-stapled-peptide-inhibitors
#15
Rike Wallbrecher, Patrick Chène, Stephan Ruetz, Therese Stachyra, Thomas Vorherr, Roland Brock
BACKGROUND AND PURPOSE: Helix stapling enhances the activity of peptides that interact with a target protein in a helical conformation. These staples are also supposed to change the pharmacokinetics of the molecules and promote cytoplasmic targeting. We scrutinized to which degree the pharmacokinetic characteristics are a function of the staple and differ from those of the standard cationic cell-penetrating peptide nonaarginine for a peptide inhibiting the interaction of p53 with the human double minute 2 (Hdm2) protein...
April 23, 2017: British Journal of Pharmacology
https://www.readbyqxmd.com/read/28435997/assessment-of-organic-pollution-of-an-industrial-river-by-synchronous-fluorescence-and-uv-vis-spectroscopy-the-fensch-river-ne-france
#16
Aziz Assaad, Steve Pontvianne, Marie-Noëlle Pons
To rapidly monitor the surface water quality in terms of organic pollution of an industrial river undergoing restoration, optical methods (UV-visible spectrometry and fluorescence) were applied in parallel to classical physical-chemical analyses. UV-visible spectra were analyzed using the maximum of the second derivative at 225 nm (related to nitrates), specific absorbance at 254 nm (SUVA254), and the spectral slope between 275 and 295 nm (S 275-295) (related to the aromaticity and molecular weight of dissolved organic carbon)...
May 2017: Environmental Monitoring and Assessment
https://www.readbyqxmd.com/read/28435908/development-of-a-japanese-encephalitis-virus-like-particle-vaccine-in-silkworms-using-codon-optimised-prm-and-envelope-genes
#17
Sayaka Matsuda, Reiko Nerome, Kenichi Maegawa, Akira Kotaki, Shigeo Sugita, Kazunori Kawasaki, Kazumichi Kuroda, Ryoji Yamaguchi, Tomohiko Takasaki, Kuniaki Nerome
We have successfully prepared a Japanese encephalitis virus (JEV) - Nakayama virus like particle (NVLP) vaccine using synthetic codon-optimized prM and E genes. The expression of the recombinant JEV Nakayama-BmNPV (JEV-NNPV) virus was determined in infected silkworm Bm-N cells by fluorescence and Western blot analysis. The recombinant was inoculated into silkworm pupae and the yield of Nakayama VLP (NVLP) reached a peak in the homogenates after 3 days. Additionally, in the peptide analysis of infected pupae homogenate, it appeared approximately 300-500 μg E protein/pupa were produced...
April 2017: Heliyon
https://www.readbyqxmd.com/read/28435892/dramatic-improvement-of-crispr-cas9-editing-in-candida-albicans-by-increased-single-guide-rna-expression
#18
Henry Ng, Neta Dean
The clustered regularly interspaced short palindromic repeat system with CRISPR-associated protein 9 nuclease (CRISPR/Cas9) has emerged as a versatile tool for genome editing in Candida albicans. Mounting evidence from other model systems suggests that the intracellular levels of single guide RNA (sgRNA) limit the efficiency of Cas9-dependent DNA cleavage. Here, we tested this idea and describe a new means of sgRNA delivery that improves previously described methods by ~10-fold. The efficiency of Cas9/sgRNA-dependent cleavage and repair of a single-copy yeast enhanced monomeric red fluorescent protein (RFP) gene was measured as a function of various parameters that are hypothesized to affect sgRNA accumulation, including transcriptional and posttranscriptional processing...
March 2017: MSphere
https://www.readbyqxmd.com/read/28435665/a%C3%A2-voltage-dependent-fluorescent-indicator-for-optogenetic-applications-archaerhodopsin-3-structure-and-optical-properties-from-in-silico-modeling
#19
Dmitrii M Nikolaev, Anton Emelyanov, Vitaly M Boitsov, Maxim S Panov, Mikhail N Ryazantsev
It was demonstrated in recent studies that some rhodopsins can be used in optogenetics as fluorescent indicators of membrane voltage. One of the promising candidates for these applications is archaerhodopsin-3. However, the fluorescent signal for wild-type achaerhodopsin-3 is not strong enough for real applications. Rational design of mutants with an improved signal is an important task, which requires both experimental and theoretical studies. Herein, we used a homology-based computational approach to predict the three-dimensional structure of archaerhodopsin-3, and a Quantum Mechanics/Molecular Mechanics (QM/MM) hybrid approach with high-level multireference ab initio methodology (SORCI+Q/AMBER) to model optical properties of this protein...
2017: F1000Research
https://www.readbyqxmd.com/read/28435614/time-resolved-biophysical-approaches-to-nucleocytoplasmic-transport
#20
REVIEW
Francesco Cardarelli
Molecules are continuously shuttling across the nuclear envelope barrier that separates the nucleus from the cytoplasm. Instead of being just a barrier to diffusion, the nuclear envelope is rather a complex filter that provides eukaryotes with an elaborate spatiotemporal regulation of fundamental molecular processes, such as gene expression and protein translation. Given the highly dynamic nature of nucleocytoplasmic transport, during the past few decades large efforts were devoted to the development and application of time resolved, fluorescence-based, biophysical methods to capture the details of molecular motion across the nuclear envelope...
2017: Computational and Structural Biotechnology Journal
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