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https://www.readbyqxmd.com/read/29779223/gene-editing-vectors-for-studying-nicotinic-acetylcholine-receptors-in-cholinergic-transmission
#1
Can Peng, Yijin Yan, Veronica J Kim, Staci E Engle, Jennifer N Berry, J Michael McIntosh, Rachael L Neve, Ryan M Drenan
Nicotinic acetylcholine receptors (nAChRs), prototype members of the cys-loop ligand gated ion channel family, are key mediators of cholinergic transmission in the central nervous system. Despite their importance, technical gaps exist in our ability to dissect the function of individual subunits in the brain. To overcome these barriers, we designed CRISPR/Cas9 small guide RNA sequences (sgRNAs) for production of loss-of-function alleles in mouse nAChR genes. These sgRNAs were validated in vitro via deep sequencing...
May 19, 2018: European Journal of Neuroscience
https://www.readbyqxmd.com/read/29779095/crispr-cas13a-targeting-of-rna-virus-in-plants
#2
Kulbhushan Chaudhary
This approach is quite promising to control plant viral diseases and create synthetic networks to better understand the structure/function relationship in RNA and proteins. Plant viruses are obligate intracellular parasites which causes enormous losses in crop yield worldwide. These viruses replicate into infected cells by highjacking host cellular machinery. Over the last two decades, diverse approaches such as conventional breeding, transgenic approach and gene silencing strategies have been used to control RNA viruses, but escaped due to high rate of mutation...
May 19, 2018: Plant Cell Reports
https://www.readbyqxmd.com/read/29778892/structure-based-design-of-novel-quinoxaline-2-carboxylic-acids-and-analogues-as-pim-1-inhibitors
#3
Bruno Oyallon, Marie Brachet-Botineau, Cédric Logé, Pascal Bonnet, Mohamed Souab, Thomas Robert, Sandrine Ruchaud, Stéphane Bach, Pascal Berthelot, Fabrice Gouilleux, Marie-Claude Viaud-Massuard, Caroline Denevault-Sabourin
We identified a new series of quinoxaline-2-carboxylic acid derivatives, targeting the human proviral integration site for Moloney murine leukemia virus-1 (HsPim-1) kinase. Seventeen analogues were synthesized providing useful insight into structure-activity relationships studied. Docking studies realized in the ATP pocket of HsPim-1 are consistent with an unclassical binding mode of these inhibitors. The lead compound 1 was able to block HsPim-1 enzymatic activity at nanomolar concentrations (IC50 of 74 nM), with a good selectivity profile against a panel of mammalian protein kinases...
May 11, 2018: European Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/29778875/purification-of-recombinant-trichodysplasia-spinulosa-associated-polyomavirus-vp1-derived-virus-like-particles-using-chromatographic-techniques
#4
Mindaugas Zaveckas, Karolis Goda, Danguole Ziogiene, Alma Gedvilaite
Trichodysplasia spinulosa-associated polyomavirus (TSPyV) has been linked to a rare and recently characterized skin disease occurring in immunocompromised patients. In analogy with other polyomaviruses, the major capsid protein VP1 of TSPyV can self-assemble into virus-like particles (VLPs). VLPs are increasingly applied for the vaccination and diagnostics. Mostly, non-scalable and labor intensive ultracentrifugation-based techniques are used for the purification of VLPs. In this work, we developed a purification procedure for TSPyV VP1 VLPs based on two chromatographic steps, ion-exchange monolith and core bead chromatography...
May 9, 2018: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/29778844/identification-of-micrornas-with-heat-stress-responsive-and-immune-properties-in-marsupenaeus-japonicus-based-on-next-generation-sequencing-and-bioinformatics-analysis-essential-regulators-in-the-heat-stress-host-interactions
#5
Jinbin Zheng, Jiawen Cao, Yong Mao, Yongquan Su, Jun Wang
Summer mortality syndrome is one of the most serious issue for Marsupenaeus japonicus aquaculture in China. Since it causes massive economic loss and threatens sustainability of M. japonicus aquaculture industry, thus, there is an urgent desire to reveal the heat stress-host interactions mechanisms that lead to mass mortalities of M. japonicus in hot summer months. MicroRNAs (miRNAs) are small noncoding RNAs that involved in regulation of diverse biological processes, including stress and immune response, and might serve as potential regulators in the heat stress-host interactions...
May 17, 2018: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/29778741/changes-in-lipid-protein-and-carbohydrate-metabolism-in-spodoptera-exigua-larvae-associated-with-infection-by-heliothis-virescens-ascovirus-3h
#6
Zi-Qi Li, Huan Yu, Guo-Hua Huang
Spodoptera exigua (Lepidoptera: Noctuidae) larvae were used to analyze the regulation of lipid, protein and carbohydrate metabolism in host larvae infected with the Heliothis virescens ascovirus 3h (HvAV-3h). Using histological sections, significant pathological changes were found in the fat bodies of infected larvae from 24 h to 72 h post-infection (hpi). The lipid and protein contents of the infected larvae were significantly higher than those of the uninfected larvae, while the carbohydrate content of the infected larvae was significantly lower than that of the mock-infected larvae...
May 17, 2018: Journal of Invertebrate Pathology
https://www.readbyqxmd.com/read/29778701/natural-history-of-avian-papillomaviruses
#7
REVIEW
Daniel A Truchado, Richard A J Williams, Laura Benítez
Papillomaviruses (Family: Papillomaviridae) are small non-enveloped viruses that cause skin and mucosa infections in diverse vertebrates. The vast majority have been detected in mammals. However, the number of papillomaviruses described in birds is growing, especially because of metagenomic studies. Seven complete genomes and one partial sequence have been described, corresponding to five papillomavirus genera. These have been detected from various sample types, including skin, internal epithelium, and faecal material, from seven highly diverse wild and captive avian species...
May 17, 2018: Virus Research
https://www.readbyqxmd.com/read/29778543/expression-of-a-functional-intrabody-against-hepatitis-c-virus-core-protein-in-escherichia-coli-and-silkworm-pupae
#8
Tatsuya Kato, Moeko Hasegawa, Takeshi Yamamoto, Takatsugu Miyazaki, Ryosuke Suzuki, Takaji Wakita, Tetsuro Suzuki, Enoch Y Park
It has been shown that the single-domain intrabody 2H9-L against the hepatitis C virus (HCV) capsid (core) protein inhibits the viral propagation and NF-κB promoter activity induced by the HCV core. In this study, 2H9-L fused with the FLAG tag sequence was expressed in both Escherichia coli and silkworm pupae and then purified. In addition, the full-length and its C terminal deletions of the HCV core protein, i.e., 1-123 amino acid residues (C123), 1-152 amino acid residues (C152), 1-177 amino acid residues (C177) and 1-191 amino acid residues (C191), were expressed as fusion proteins with a 6 × His tag at their N-terminus in E...
May 17, 2018: Protein Expression and Purification
https://www.readbyqxmd.com/read/29778203/evaluation-of-the-immune-response-afforded-by-a-subunit-vaccine-candidate-against-bluetongue-virus-in-mice-and-sheep
#9
Darien Kheder Ali Mohamed, Junzheng Du, Shandian Gao, Zhancheng Tian, Guorui Zhang, Dexuan Huang, Rongsheng Du, Biao Kang, Guangyuan Liu, Jianxun Luo, Hong Yin
Bluetongue virus (BTV), a vector-borne pathogen, is the causative agent of bluetongue disease in ruminants. In view of the recent emergence of BTV in regions previously known to be free from the disease and/or specific serotypes or strains, optimization of the currently available vaccination strategies to control the spread of vector-borne bluetongue is crucial. The main objective of the current study was to develop a subunit vaccine candidate targeting BTV-16, a strain previously isolated in China from sheep with obvious clinical signs...
June 2018: Veterinary Microbiology
https://www.readbyqxmd.com/read/29778202/porcine-monocyte-derived-dendritic-cells-can-be-differentially-activated-by-vesicular-stomatitis-virus-and-its-matrix-protein-mutants
#10
Guoyan Sun, Xinkui Fang, Hao Wu, Xinchu Zhou, Yong Ke, Tao Sun
Vesicular stomatitis virus (VSV) can cause serious vesicular lesions in pigs, and the matrix (M) protein is its predominant virulence factor. Dendritic cells (DCs) act as the bridge between innate and adaptive immune responses. However, the susceptibility of porcine DCs to VSV infection and the role of M protein in modulating the function of infected DCs are still poorly defined. Thus, this study aimed to determine the ability of virulent wild-type VSV(wtVSV) and two attenuated M protein variants (VSVΔM51 and VSVMT ) to induce maturation of porcine monocyte-derived DCs (MoDCs) in vitro...
June 2018: Veterinary Microbiology
https://www.readbyqxmd.com/read/29778186/identification-of-antigenic-epitopes-of-monoclonal-antibodies-against-the-vp2-protein-of-the-25-serotype-of-bluetongue-virus
#11
Shuangyu Xie, Yiming Shi, Ruyue Gong, Wen Cui, Yanping Jiang, Min Liu, Li Wang, Han Zhou, Xinyuan Qiao, Yijing Li, Yigang Xu, Lijie Tang
Serious outbreaks of bluetongue, an arbovirus of domestic and wild ruminants caused by bluetongue virus serotypes (BTV), have occurred around the world. More than 27 distinct serotypes are recognized throughout the world. A new virus, BTV-25 (Toggenburg orbivirus [TOV]), was first detected in Switzerland, and has not yet been found in China. VP2 is an important outer shell protein that defines BTV serotypes and is, therefore, an ideal target antigen for serotype identification. To produce a monoclonal antibody against VP2 of BTV-25, the segment 2 gene was divided into three segments, cloned into pET-28a (+) and pMAL-c5X vectors, and the protein was expressed in E...
June 2018: Veterinary Microbiology
https://www.readbyqxmd.com/read/29778185/serine-105-and-120-are-important-phosphorylation-sites-for-porcine-reproductive-and-respiratory-syndrome-virus-n-protein-function
#12
Yao Chen, Xiulin Xing, Qi Li, Songlin Feng, Xiaoliang Han, Shuyi He, Guihong Zhang
The nucleocapsid (N) protein is the most abundant protein of porcine reproductive and respiratory syndrome virus (PRRSV). It has been shown to be multiphosphorylated. However, the phosphorylation sites are still unknown. In this study, we used liquid chromatography tandem mass spectrometry (LC-MS/MS) to analyze the phosphorylation sites of N protein expressed in Sf9 cells. The results showed that N protein contains two phosphorylation sites. Since N protein can regulate IL-10, which may facilitate PRRSV replication, we constructed four plasmids (pCA-XH-GD, pCA-A105, pCA-A120 and pCA-A105-120) and transfected them into Pig alveolar macrophages (PAMs,3D4/2)...
June 2018: Veterinary Microbiology
https://www.readbyqxmd.com/read/29778179/recombinant-adenovirus-delivered-soluble-cd163-and-sialoadhesin-receptors-protected-pigs-from-porcine-reproductive-and-respiratory-syndrome-virus-infection
#13
Wenlong Xia, Zhi Wu, Changming Guo, Shanyuan Zhu, Xinyu Zhang, Xiaoli Xia, Huaichang Sun
Porcine reproductive and respiratory syndrome (PRRS) is one of the most important swine diseases affecting pig industry worldwide. Sialoadehesin (Sn) and CD163 are the two specific receptors for PRRSV infection of porcine alveolar macrophages. Our previous study showed that the soluble Sn receptor Sn4D-Fc and soluble CD163 receptor SRCR59-Fc expressed by the two recombinant adenoviral (rAd) vectors have an additive anti-PRRSV effect in vitro. In the present study, rAd-Sn4D-Fc and rAd-SRCR59-Fc were inoculated into pigs, and the efficient expression of Sn4D-Fc and SRCR59-Fc proteins was detected by ELISA...
June 2018: Veterinary Microbiology
https://www.readbyqxmd.com/read/29777913/the-tp53-induced-glycolysis-and-apoptosis-regulator-mediates-cooperation-between-htlv-1-p30-ii-and-the-retroviral-oncoproteins-tax-and-hbz-and-is-highly-expressed-in-an-in-vivo-xenograft-model-of-htlv-1-induced-lymphoma
#14
Tetiana Hutchison, Aditi Malu, Laçin Yapindi, Rachel Bergeson, Kendra Peck, Megan Romeo, Carolyn Harrod, Jordan Pope, Louisa Smitherman, Wesleigh Gwinn, Lee Ratner, Courtney Yates, Robert Harrod
The human T-cell leukemia virus type-1 (HTLV-1) is an oncoretrovirus that infects and transforms CD4+ T-cells and causes adult T-cell leukemia/lymphoma (ATLL) -an aggressive lymphoproliferative disease that is highly refractive to most anticancer therapies. The HTLV-1 proviral genome encodes several regulatory products within a conserved 3' nucleotide sequence, known as pX; however, it remains unclear how these factors might cooperate or dynamically interact in virus-infected cells. Here we demonstrate that the HTLV-1 latency-maintenance factor p30II induces the TP53-induced glycolysis and apoptosis regulator (TIGAR) and counters the oxidative stress, mitochondrial damage, and cytotoxicity caused by the viral oncoproteins Tax and HBZ...
May 16, 2018: Virology
https://www.readbyqxmd.com/read/29777875/switching-cell-penetrating-and-cxcr4-binding-activities-of-nanoscale-organized-arginine-rich-peptides
#15
Marianna Teixeira de Pinho Favaro, Naroa Serna, Laura Sánchez-García, Rafael Cubarsi, Mónica Roldán, Alejandro Sánchez-Chardi, Ugutz Unzueta, Ramón Mangues, Neus Ferrer-Miralles, Adriano Rodrigues Azzoni, Esther Vázquez, Antonio Villaverde
Arginine-rich protein motifs have been described as potent cell-penetrating peptides (CPPs) but also as rather specific ligands of the cell surface chemokine receptor CXCR4, involved in the infection by the human immunodeficiency virus (HIV). Polyarginines are commonly used to functionalize nanoscale vehicles for gene therapy and drug delivery, aimed to enhance cell penetrability of the therapeutic cargo. However, under which conditions these peptides do act as either unspecific or specific ligands is unknown...
May 16, 2018: Nanomedicine: Nanotechnology, Biology, and Medicine
https://www.readbyqxmd.com/read/29777870/profiling-modifications-for-glioblastoma-proteome-using-ultra-tolerant-database-search-are-the-peptide-mass-shifts-biologically-relevant-or-chemically-induced
#16
Irina A Tarasova, Peter M Chumakov, Sergei A Moshkovskii, Mikhail V Gorshkov
Peptide mass shifts were profiled using ultra-tolerant database search strategy for shotgun proteomics data sets of human glioblastoma cell lines demonstrating strong response to the type I interferon (IFNα-2b) treatment. The main objective of this profiling was revealing the cell response to IFN treatment at the level of protein modifications. To achieve this objective, statistically significant changes in peptide mass shift profiles between IFN treated and untreated glioblastoma samples were analyzed. Detailed analysis of MS/MS spectra allowed further interpretation of the observed mass shifts and differentiation between post-translational and artifact modifications...
May 16, 2018: Journal of Proteomics
https://www.readbyqxmd.com/read/29777803/an-isolate-of-potato-virus-x-capsid-protein-from-n-benthamiana-insights-from-homology-modeling-and-molecular-dynamics-simulation
#17
Neda Esfandiari, Yahya Sefidbakht
Since Potato Virus X (PVX) is easily transmitted mechanically between their hosts, its control is difficult. We have previously reported new isolate of this virus (PVX-Iran, GenBank Accession number FJ461343). However, the molecular basis of resistance breaking activity and its relation to capsid protein structure are still not well-understood. SDS-PAGE, ELISA, Western blot and RT-PCR molecular examinations were performed on the inoculated plants Nicotiana benthamiana. The pathological symptoms were related to the PVX isolate...
May 16, 2018: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/29777766/immune-response-and-protective-effect-against-spring-viremia-of-carp-virus-induced-by-intramuscular-vaccination-with-a-swcnts-dna-vaccine-encoding-matrix-protein
#18
Chen Zhang, Zhao Zhao, Gao-Yang Liu, Jian Li, Gao-Xue Wang, Bin Zhu
To elicit the immune protective of vaccine against the highly contagious and pathogenic disease caused by spring viremia of carp virus (SVCV), a novel functionalized single-walled carbon nanotubes (SWCNTs) were applied as a delivery vehicle for DNA vaccine. In this study, we report a SWCNTs-DNA vaccine encoding matrix protein of SVCV which, when injected in the muscle at a dose of 10 μg SWCNTs-pcDNA-M vaccine, confers up to 51.3% protection against intraperitoneal challenge with SVCV. In addition, SWCNTs as a promising vehicle can enhance about 17...
May 16, 2018: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/29777657/mcv-mir-m1-targets-the-host-cell-immune-response-resulting-in-the-attenuation-of-neutrophil-chemotaxis
#19
P Akhbari, D Tobin, K Poterlowicz, W Roberts, J R Boyne
Virus-encoded miRNAs are emerging as key regulators of persistent infection and host-cell immune evasion. Merkel cell polyomavirus (MCPyV), the predominant aetiological agent of Merkel cell carcinoma (MCC), encodes a single miRNA, MCV-miR-M1, which targets the oncogenic MCPyV large T antigen (LT). MCV-miR-M1 has previously been shown to play an important role in establishment of long-term infection, however, the underlying mechanism is not fully understood. A key unanswered question is whether, in addition to auto-regulating LT, MCV-miR-M1 also targets cellular transcripts to orchestrate an environment conducive for persistent infection...
May 16, 2018: Journal of Investigative Dermatology
https://www.readbyqxmd.com/read/29777503/anti-tmv-effects-of-amaryllidaceae-alkaloids-isolated-from-the-bulbs-of-lycoris-radiata-and-lycoricidine-derivatives
#20
Dong-Qiong Yang, Zhao-Rong Chen, Duo-Zhi Chen, Xiao-Jiang Hao, Shun-Lin Li
Fifteen known amaryllidaceae alkaloids were isolated from the bulbs of Lycoris radiata. Some of the compounds and lycoricidine derivatives had been screened for the activities against tobacco mosaic virus (TMV) by the conventional half-leaf method. Lycoricidine derivatives were also carried out the assay of effect on systemic infection of TMV by western-blot and RT-PCR analysis. The tested compounds showed moderate inactivation effect, whereas the lycoricidine derivatives showed good protective effect. The protective inhibitory activity of compounds L1 (N-methyl-2,3,4-trimethoxylycoricidine) (60...
May 18, 2018: Natural Products and Bioprospecting
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