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https://www.readbyqxmd.com/read/29235978/rigid-amphipathic-fusion-inhibitors-demonstrate-antiviral-activity-against-african-swine-fever-virus
#1
Astghik Hakobyan, Inmaculada Galindo, Almudena Nañez, Erik Arabyan, Zaven Karalyan, Alexey A Chistov, Philipp P Streshnev, Vladimir A Korshun, Covadonga Alonso, Hovakim Zakaryan
Rigid amphipathic fusion inhibitors (RAFIs) are a family of nucleoside derivatives that inhibit the infectivity of several enveloped viruses by interacting with virion envelope lipids and inhibiting fusion between viral and cellular membranes. Here we tested the antiviral activity of two RAFIs, 5-(Perylen-3-ylethynyl)-arabino-uridine (aUY11) and 5-(Perylen-3-ylethynyl)uracil-1-acetic acid (cm1UY11) against African swine fever virus (ASFV), for which no effective vaccine is available. Both compounds displayed a potent, dose-dependent inhibitory effect on ASFV infection in Vero cells...
December 13, 2017: Journal of General Virology
https://www.readbyqxmd.com/read/29235215/cloning-free-template-dna-preparation-for-cell-free-protein-synthesis-via-two-step-pcr-using-versatile-primer-designs-with-short-3-utr
#2
Mika Nomoto, Yasuomi Tada
Cell-free protein synthesis (CFPS) systems largely retain the endogenous translation machinery of the host organism, making them highly applicable for proteomics analysis of diverse biological processes. However, laborious and time-consuming cloning procedures hinder progress with CFPS systems. Herein, we report the development of a rapid and efficient two-step polymerase chain reaction (PCR) method to prepare linear DNA templates for a wheat germ CFPS system. We developed a novel, effective short 3'-untranslated region (3'-UTR) sequence that facilitates translation...
December 13, 2017: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
https://www.readbyqxmd.com/read/29235053/classification-of-c2c12-cells-at-differentiation-by-convolutional-neural-network-of-deep-learning-using-phase-contrast-images
#3
Hirohiko Niioka, Satoshi Asatani, Aina Yoshimura, Hironori Ohigashi, Seiichi Tagawa, Jun Miyake
In the field of regenerative medicine, tremendous numbers of cells are necessary for tissue/organ regeneration. Today automatic cell-culturing system has been developed. The next step is constructing a non-invasive method to monitor the conditions of cells automatically. As an image analysis method, convolutional neural network (CNN), one of the deep learning method, is approaching human recognition level. We constructed and applied the CNN algorithm for automatic cellular differentiation recognition of myogenic C2C12 cell line...
December 13, 2017: Human Cell
https://www.readbyqxmd.com/read/29234687/delivery-of-cr2-fh-using-aav-vector-therapy-as-treatment-strategy-in-the-mouse-model-of-choroidal-neovascularization
#4
Gloriane Schnabolk, Nathaniel Parsons, Elisabeth Obert, Balasubramaniam Annamalai, Cecile Nasarre, Stephen Tomlinson, Alfred S Lewin, Bärbel Rohrer
Complement activation plays a significant role in age-related macular degeneration (AMD) pathogenesis, and polymorphisms interfering with factor H (fH) function, a complement alternative pathway (AP) inhibitor, are associated with increased AMD risk. We have previously validated an AP inhibitor, a fusion protein consisting of a complement receptor 2 fragment linked to the inhibitory domain of fH (CR2-fH) as an efficacious treatment for choroidal neovascularization (CNV) when delivered intravenously. Here we tested an alternative approach of AAV-mediated delivery (AAV5-VMD2-CR2-fH or AAV5-VMD2-mCherry) using subretinal delivery in C57BL/6J mice...
June 15, 2018: Molecular Therapy. Methods & Clinical Development
https://www.readbyqxmd.com/read/29234347/ectopic-expression-of-pumpkin-nac-transcription-factor-cmnac1-improves-multiple-abiotic-stress-tolerance-in-arabidopsis
#5
Haishun Cao, Li Wang, Muhammad A Nawaz, Mengliang Niu, Jingyu Sun, Junjun Xie, Qiusheng Kong, Yuan Huang, Fei Cheng, Zhilong Bie
Drought, cold and salinity are the major environmental stresses that limit agricultural productivity. NAC transcription factors regulate the stress response in plants. Pumpkin (Cucurbita moschata) is an important cucurbit vegetable crop and it has strong resistance to abiotic stress; however, the biological functions of stress-related NAC genes in this crop are largely unknown. This study reports the function of CmNAC1, a stress-responsive pumpkin NAC domain protein. The CmNAC1-GFP fusion protein was transiently expressed in tobacco leaves for subcellular localization analysis, and we found that CmNAC1 is localized in the nucleus...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/29234103/an-exhaustive-search-algorithm-to-aid-nmr-based-structure-determination-of-rotationally-symmetric-transmembrane-oligomers
#6
Jing Yang, Alessandro Piai, Hong-Bin Shen, James J Chou
Nuclear magnetic resonance (NMR) has been an important source of structural restraints for solving structures of oligomeric transmembrane domains (TMDs) of cell surface receptors and viral membrane proteins. In NMR studies, oligomers are assembled using inter-protomer distance restraints. But, for oligomers that are higher than dimer, these distance restraints all have two-fold directional ambiguity, and resolving such ambiguity often requires time-consuming trial-and-error calculations using restrained molecular dynamics (MD) with simulated annealing (SA)...
December 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29233896/retargeting-lentiviruses-via-spycatcher-spytag-chemistry-for-gene-delivery-into-specific-cell-types
#7
Nagarjun Kasaraneni, Ana M Chamoun-Emanuelli, Gus Wright, Zhilei Chen
We report a simple strategy for the creation of lentiviral vectors specific to any desired target cells. SpyTag is inserted into an engineered Sindbis virus envelope protein and displayed on the lentivirus surface to create Sindbis virus-SpyTag pseudoparticles (Sind-SpyTag-pp). The SpyTag serves as the covalent anchoring site for a target-cell-specific cell-binding protein (CBP) that is fused to a truncated SpyCatcher (SpyCatcherΔ). Target-cell-specific lentiviruses are created by mixing the Sind-SpyTag-pp and CBP-SpyCatcherΔ in vitro We first used a HER2-binding designed ankyrin repeat protein (DARPin...
December 12, 2017: MBio
https://www.readbyqxmd.com/read/29233140/selection-characterization-and-thermal-stabilization-of-llama-single-domain-antibodies-towards-ebola-virus-glycoprotein
#8
Jinny L Liu, Lisa C Shriver-Lake, George P Anderson, Dan Zabetakis, Ellen R Goldman
BACKGROUND: A key advantage of recombinant antibody technology is the ability to optimize and tailor reagents. Single domain antibodies (sdAbs), the recombinantly produced variable domains derived from camelid and shark heavy chain antibodies, provide advantages of stability and solubility and can be further engineered to enhance their properties. In this study, we generated sdAbs specific for Ebola virus envelope glycoprotein (GP) and increased their stability to expand their utility for use in austere locals...
December 12, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/29232933/a-split-luciferase-reporter-recognizing-gfp-and-mcherry-tags-to-facilitate-studies-of-protein-protein-interactions
#9
Mehdi Moustaqil, Akshay Bhumkar, Laura Gonzalez, Lisa Raoul, Dominic J B Hunter, Pascal Carrive, Emma Sierecki, Yann Gambin
The use of fluorescently-tagged proteins in microscopy has become routine, and anti-GFP (Green fluorescent protein) affinity matrices are increasingly used in proteomics protocols. However, some protein-protein interactions assays, such as protein complementation assays (PCA), require recloning of each protein as a fusion with the different parts of the complementation system. Here we describe a generic system where the complementation is separated from the proteins and can be directly used with fluorescently-tagged proteins...
December 11, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29231808/abelson-tyrosine-protein-kinase-2-regulates-myoblast-proliferation-and-controls-muscle-fiber-length
#10
Jennifer K Lee, Steven J Burden
Muscle fiber length is nearly uniform within a muscle but widely different among different muscles. We show that Abelson tyrosine-protein kinase 2 (Abl2) has a key role in regulating myofiber length, as a loss of Abl2 leads to excessively long myofibers in the diaphragm, intercostal and levator auris muscles but not limb muscles. Increased myofiber length is caused by enhanced myoblast proliferation, expanding the pool of myoblasts and leading to increased myoblast fusion. Abl2 acts in myoblasts, but as a consequence of expansion of the diaphragm muscle, the diaphragm central tendon is reduced in size, likely contributing to reduced stamina of Abl2 mutant mice...
December 12, 2017: ELife
https://www.readbyqxmd.com/read/29231480/-effects-of-electroacupuncture-on-synaptic-plasticity-in-hippocampal-ca3-area-of-rats-with-chronic-stress-depression
#11
Long Wang, Di Zhang, Xusheng Tian, Dan Liu, Xiaowei Sun, Yanqiu Li, Cuijuan Wang, Ming Ma, Dongjie Li, Zhongren Sun
OBJECTIVE: To explore the effects of electroacupuncture (EA) on behavioral function and synaptic plasticity in hippocampal CA3 area in rats with chronic stress depression. METHODS: According to the random number table method, 144 SD male rats were assigned into a blank group, a model group, an EA group and a fluoxetine group, then each group was divided into a 7 d subgroup, a 14 d subgroup and a 21 d subgroup, 12 rats in each subgroup. The chronic mild unpredictable stress stimulus combined with lonely breeding were applied to establish the depression model of rats, which was performed simultaneously with intervention treatment...
February 12, 2017: Zhongguo Zhen Jiu, Chinese Acupuncture & Moxibustion
https://www.readbyqxmd.com/read/29230547/novel-fluorochromes-label-tonoplast-in-living-plant-cells-and-reveal-changes-in-vacuolar-organization-after-treatment-with-protein-phosphatase-inhibitors
#12
Miklós Nagy, Sándor Kéki, Dávid Rácz, Jaideep Mathur, György Vereb, Tamás Garda, Márta M-Hamvas, François Chaumont, Károly Bóka, Béla Böddi, Csongor Freytag, Gábor Vasas, Csaba Máthé
The recently synthesized isocyanonaphtalene derivatives ACAIN and CACAIN are fluorochromes excitable at wavelengths of around 366 nm and bind cysteine-rich proteins with hydrophobic motifs. We show that these compounds preferentially label tonoplasts in living Arabidopsis and tobacco (Nicotiana tabacum SR1) cells. ACAIN-labeled membranes co-localized with the GFP signal in plants expressing GFP-δ-TIP (TIP2;1) (a tonoplast aquaporin) fusion protein. ACAIN preserved the dynamics of vacuolar structures. tip2;1 and triple tip1;1-tip1;2-tip2;1 knockout mutants showed weaker ACAIN signal in tonoplasts...
December 12, 2017: Protoplasma
https://www.readbyqxmd.com/read/29230372/integrated-downstream-regulation-by-the-quorum-sensing-controlled-transcription-factors-lrha-and-rcsa-impacts-phenotypic-outputs-associated-with-virulence-in-the-phytopathogen-pantoea-stewartii-subsp-stewartii
#13
Duy An Duong, Ann M Stevens
Pantoea stewartii subsp. stewartii is a Gram-negative proteobacterium that causes leaf blight and Stewart's wilt disease in corn. Quorum sensing (QS) controls bacterial exopolysaccharide production that blocks water transport in the plant xylem at high bacterial densities during the later stage of the infection, resulting in wilt. At low cell density the key master QS regulator in P. stewartii, EsaR, directly represses rcsA, encoding an activator of capsule biosynthesis genes, but activates lrhA, encoding a transcription factor that regulates surface motility...
2017: PeerJ
https://www.readbyqxmd.com/read/29230211/evaluation-of-aggregated-ag85b-antigen-for-its-biophysical-properties-immunogenicity-and-vaccination-potential-in-a-murine-model-of-tuberculosis-infection
#14
Faraz Ahmad, Swaleha Zubair, Pushpa Gupta, Umesh Datta Gupta, Rakesh Patel, Mohammad Owais
Protein aggregates have been reported to act as a reservoir that can release biologically active, native form of precursor protein. Keeping this fact into consideration, it is tempting to exploit protein aggregate-based antigen delivery system as a functional vaccine to expand desirable immunological response in the host. Herein, we explored the capacity of aggregated Ag85B of Mycobacterium tuberculosis (Mtb) to act as a prophylactic vaccine system that releases the precursor antigen in slow and sustained manner...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/29230043/nanoparticle-orientation-to-control-rna%C3%A2-loading-and-ligand-display-on-extracellular-vesicles-for-cancer-regression
#15
Fengmei Pi, Daniel W Binzel, Tae Jin Lee, Zhefeng Li, Meiyan Sun, Piotr Rychahou, Hui Li, Farzin Haque, Shaoying Wang, Carlo M Croce, Bin Guo, B Mark Evers, Peixuan Guo
Nanotechnology offers many benefits, and here we report an advantage of applying RNA nanotechnology for directional control. The orientation of arrow-shaped RNA was altered to control ligand display on extracellular vesicle membranes for specific cell targeting, or to regulate intracellular trafficking of small interfering RNA (siRNA) or microRNA (miRNA). Placing membrane-anchoring cholesterol at the tail of the arrow results in display of RNA aptamer or folate on the outer surface of the extracellular vesicle...
December 11, 2017: Nature Nanotechnology
https://www.readbyqxmd.com/read/29229733/expression-of-tlr2-by-dendritic-cells-is-essential-for-dnaj-%C3%AE-a146ply-mediated-th1-immune-response-against-streptococcus-pneumoniae
#16
Xiaofang Wang, Taixian Yuan, Jun Yuan, Yufeng Su, Xiaoyu Sun, Jingwen Wu, Hong Zhang, Xun Min, Xuemei Zhang, Yibing Yin
The fusion protein, DnaJ-ΔA146Ply, could induce cross-protective immunity against pneumococcal infection via mucosal and subcutaneous immunization in mice in the absence of additional adjuvants. DnaJ and Ply are both TLR4 but not TLR2 ligands. However, we found that TLR2-/- mice immunized subcutaneously with DnaJ-ΔA146Ply showed significantly lower survival rates and higher bacterial loads in nasal washes than those of WT mice after being challenged with pneumococcal strains D39 or 19F. The IFN-γ level in splenocytes decreased in TLR2-/- mice, indicating that Th1 immunity elicited by DnaJ-ΔA146Ply was impaired in TLR2-/- mice...
December 11, 2017: Infection and Immunity
https://www.readbyqxmd.com/read/29229732/utilization-of-variant-and-fusion-proteins-to-functionally-map-the-aggregatibacter-actinomycetemcomitans-trimeric-autotransporter-protein-apia
#17
Carla Cugini, Yongyi Mei, David Furgang, Nisha George, Narayanan Ramasubbu, Daniel H Fine
The Gram-negative bacterium Aggregatibacter actinomycetemcomitans is a causative agent of localized aggressive periodontitis. Critical to its infection process is the first and essential step of attachment, which is related to the coordinated functions of surface components comprised of proteins and extracellular polysaccharides. One such protein is the outer membrane trimeric autotransporter protein ApiA, a versatile virulence factor with numerous functions including cell binding, invasion, serum resistance, autoaggregation, and induction of cytokine release...
December 11, 2017: Infection and Immunity
https://www.readbyqxmd.com/read/29229583/integrative-genomic-analysis-of-coincident-cancer-foci-implicates-ctnnb1-and-pten-alterations-in-ductal-prostate-cancer
#18
Marc Gillard, Justin Lack, Andrea Pontier, Divya Gandla, David Hatcher, Adam G Sowalsky, Jose Rodriguez-Nieves, Donald Vander Griend, Gladell Paner, David VanderWeele
BACKGROUND: Ductal adenocarcinoma of the prostate is an aggressive subtype, with high rates of biochemical recurrence and overall poor prognosis. It is frequently found coincident with conventional acinar adenocarcinoma. The genomic features driving evolution to its ductal histology and the biology associated with its poor prognosis remain unknown. OBJECTIVE: To characterize genomic features distinguishing ductal adenocarcinoma from coincident acinar adenocarcinoma foci from the same patient...
December 8, 2017: European Urology Focus
https://www.readbyqxmd.com/read/29229552/caspase-6-is-a-dispensable-enabler-of-adult-mammalian-axonal-degeneration
#19
V Woo, C Cheng, A Duraikannu, A Chandrasekhar, K Purdy, J A Martinez, D W Zochodne
The progress of axonal degeneration (AxD) following injury or insult impacts both recovery from axonal transection and protection of axons from diverse insults, or axonopathy. Here we provide evidence that increases in capase-6 (Casp6) expression and action contribute to the progression of AxD. The expression of Casp6 protein and mRNA in distal branches of sensory axons undergoing AxD was confirmed. We developed and utilized a new model of axonopathy in live mice by serially visualizing the viability of cutaneous axons in the ear pinna that expressed an axonal YFP transgene, in response to capasaicin-induced AxD...
December 8, 2017: Neuroscience
https://www.readbyqxmd.com/read/29229290/simple-detection-of-telomere-fusions-in-pancreatic-cancer-intraductal-papillary-mucinous-neoplasm-and-pancreatic-cyst-fluid
#20
Tatsuo Hata, Marco Dal Molin, Anne McGregor-Das, Tae Jun Song, Christopher Wolfgang, James R Eshleman, Ralph H Hruban, Michael Goggins
Telomere end-to-end fusions are an important source of chromosomal instability that arise in cells with critically shortened telomeres. We developed a nested real-time quantitative PCR method for telomere fusion detection in pancreatic ductal adenocarcinomas, intraductal papillary mucinous neoplasms (IPMNs), and IPMN cyst fluids. Ninety-one pancreatic cancer cell lines and xenograft samples, 93 IPMNs, and 93 surgically aspirated IPMN cyst fluid samples were analyzed. The association between telomere shortening, telomerase activity, and telomere fusion detection was evaluated...
December 6, 2017: Journal of Molecular Diagnostics: JMD
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