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https://www.readbyqxmd.com/read/28442010/-effect-of-nm23-h1-nuclear-localization-on-proliferation-of-%C3%A2-human-lung-adenocarcinoma-cell-line-a549
#1
Ya Sheng, Yanli Xiong, Mingfang Xu, Xunjie Kuang, Dong Wang, Xueqin Yang
BACKGROUND: Recent studies have indicated that Nm23-H1 is found in the nucleus, but previous studies have been based on the overexpression or suppression of Nm23-H1 in the cytoplasm. Due to the lacking nuclear localization signal of Nm23-H1, these results cannot reflect or repeat cells in which Nm23-H1 mainly positioned in nuclei and whether they cause clinical biological effects. Therefore, to explore the effects of transposing Nm23-H1 from the cytoplasm to the nucleus during lung cancer cell proliferation, a vector with a nuclear localization signal of Nm23-H1 was constructed and A549 cells were transfected...
April 20, 2017: Zhongguo Fei Ai za Zhi, Chinese Journal of Lung Cancer
https://www.readbyqxmd.com/read/28427998/nitric-oxide-regulated-proteolysis-of-human-cyp2b6-via-the-ubiquitin-proteasome-system
#2
Choon-Myung Lee, Shweta Tripathi, Edward T Morgan
We showed previously that rat cytochrome P450 CYP2B1 undergoes NO-dependent proteasomal degradation in response to inflammatory stimuli, and that the related human enzyme CYP2B6 is also down-regulated by NO in primary human hepatocytes. To investigate the mechanism of CYP2B6 down-regulation, we made several cell lines (HeLa and HuH7 cells) in which native CYP2B6 or CYP2B6 with a C-terminal V5 tag (CYP2B6V5) are expressed from a lentiviral vector with a cytomegalovirus promoter. Native CYP2B6 protein was rapidly down-regulated in HeLa cells within 3h of treatment with the NO donor (Z)-1-[2-(2-Aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate, while its mRNA level was not down-regulated...
April 17, 2017: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/28420741/new-mechanism-by-which-human-cytomegalovirus-micrornas-negate-the-proinflammatory-response-to-infection
#3
Andrew D Yurochko
Viruses have evolved many novel mechanisms to promote infection and to mitigate the host cell response to that infection. In the article by M. H. Hancock et al. (mBio 8:e00109-17, 2017, https://doi.org/10.1128/mBio.00109-17), the authors describe a new mechanism by which human cytomegalovirus (HCMV) microRNAs (miRNAs; miR-US5-1 and miR-UL112-3p) negate the proinflammatory response to infection. The authors document that these two viral miRNAs downregulate the NF-κB response through direct targeting of the IKKα and IKKβ mRNAs, which in turn, through diminished IκB kinases (IKKs), block production of proinflammatory cytokines (interleukin-6 [IL-6], CCL5, and tumor necrosis factor alpha [TNF-α])...
April 18, 2017: MBio
https://www.readbyqxmd.com/read/28420570/herpesviradae-infections-in-severely-burned-children
#4
Paul Wurzer, Megan R Cole, Robert P Clayton, Gabriel Hundeshagen, Omar Nunez Lopez, Janos Cambiaso-Daniel, Raimund Winter, Ludwik K Branski, Hal K Hawkins, Celeste C Finnerty, David N Herndon, Jong O Lee
OBJECTIVE: Burn-related immunosuppression can promote human herpesviridae infections. However, the effect of these infections on morbidity and mortality after pediatric burn injuries is unclear. METHODS: We retrospectively analyzed pediatric patients with burns ≥10% of the total body surface area (TBSA) who were admitted between 2010 and 2015. On clinical suspicion of a viral infection, antiviral therapy was initiated. Viral infection was confirmed via Tzanck smear, viral culture, and/or PCR...
April 15, 2017: Burns: Journal of the International Society for Burn Injuries
https://www.readbyqxmd.com/read/28411277/long-term-survival-in-glioblastoma-with-cytomegalovirus-pp65-targeted-vaccination
#5
Kristen A Batich, Elizabeth A Reap, Gary E Archer, Luis Sanchez-Perez, Smita K Nair, Robert J Schmittling, Pam Norberg, Weihua Xie, James E Herndon, Patrick Healy, Roger E McLendon, Allan H Friedman, Henry S Friedman, Darell Bigner, Gordana Vlahovic, Duane A Mitchell, John H Sampson
Purpose: Patients with glioblastoma have less than 15-month median survival despite surgical resection, high-dose radiation, and chemotherapy with temozolomide. We previously demonstrated that targeting cytomegalovirus pp65 using dendritic cells (DC) can extend survival and, in a separate study, that dose-intensified temozolomide (DI-TMZ) and adjuvant granulocyte macrophage colony-stimulating factor (GM-CSF) potentiate tumor-specific immune responses in patients with glioblastoma. Here, we evaluated pp65-specific cellular responses following DI-TMZ with pp65-DCs and determined the effects on long-term progression-free survival (PFS) and overall survival (OS)...
April 15, 2017: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/28409504/comparative-study-of-the-transfection-efficiency-of-commonly-used-viral-vectors-in-rhesus-monkey-macaca-mulatta-brains
#6
Shi-Hao Wu, Zhi-Xing Liao, Joshua D Rizak, Na Zheng, Lin-Heng Zhang, Hen Tang, Xiao-Bin He, Yang Wu, Xia-Ping He, Mei-Feng Yang, Zheng-Hui Li, Dong-Dong Qin, Xin-Tian Hu
Viral vector transfection systems are among the simplest of biological agents with the ability to transfer genes into the central nervous system. In brain research, a series of powerful and novel gene editing technologies are based on these systems. Although many viral vectors are used in rodents, their full application has been limited in non-human primates. To identify viral vectors that can stably and effectively express exogenous genes within non-human primates, eleven commonly used recombinant adeno-associated viral and lentiviral vectors, each carrying a gene to express green or red fluorescence, were injected into the parietal cortex of four rhesus monkeys...
March 18, 2017: Zoological Research
https://www.readbyqxmd.com/read/28407741/the-pd-1-pd-l1-pathway-promotes-development-of-brain-resident-memory-t-cells-following-acute-viral-encephalitis
#7
Sujata Prasad, Shuxian Hu, Wen S Sheng, Priyanka Chauhan, Amar Singh, James R Lokensgard
BACKGROUND: Previous work from our laboratory has demonstrated that during acute viral brain infection, glial cells modulate antiviral T cell effector responses through the PD-1: PD-L1 pathway, thereby limiting the deleterious consequences of unrestrained neuroinflammation. Here, we evaluated the PD-1: PD-L1 pathway in development of brain-resident memory T cells (bTRM) following murine cytomegalovirus (MCMV) infection. METHODS: Flow cytometric analysis of immune cells was performed at 7, 14, and 30 days post-infection (dpi) to assess the shift of brain-infiltrating CD8(+) T cell populations from short-lived effector cells (SLEC) to memory precursor effector cells (MPEC), as well as generation of bTRMs...
April 13, 2017: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/28404844/intramuscular-immunization-of-mice-with-the-live-attenuated-herpes-simplex-virus-type-1-hsv-1-vaccine-strain-vc2-expressing-the-equine-herpesvirus-1-ehv-1-glycoprotein-d-gd-generates-anti-ehv-1-immune-responses-in-mice
#8
Shiliang A Liu, Brent A Stanfield, Vladimir N Chouljenko, Shan Naidu, Ingeborg Langohr, Fabio Del Piero, Jacqueline Ferracone, Alma A Roy, Konstantin G Kousoulas
Vaccination remains the best option to combat EHV-1 infection and several different strategies of vaccination have been investigated and developed over the past decades. Herein, we report that the live-attenuated Herpes Simplex Virus Type-1 (HSV-1) VC2 vaccine strain, which has been shown to be unable to enter into neurons and establish latency in mice, can be utilized as a vector for the heterologous expression of the Equine Herpesvirus-1 (EHV-1) glycoprotein D (gD), and that intramuscular immunization of mice resulted in strong anti-viral humoral and cellular immune responses...
April 12, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28403202/human-cytomegalovirus-glycoprotein-complex-gh-gl-go-uses-pdgfr-%C3%AE-as-a-key-for-entry
#9
Yiquan Wu, Adrian Prager, Simone Boos, Moritz Resch, Ilija Brizic, Michael Mach, Sabrina Wildner, Laura Scrivano, Barbara Adler
Herpesvirus gH/gL envelope glycoprotein complexes are key players in virus entry as ligands for host cell receptors and by promoting fusion of viral envelopes with cellular membranes. Human cytomegalovirus (HCMV) has two alternative gH/gL complexes, gH/gL/gO and gH/gL/UL128,130,131A which both shape the HCMV tropism. By studying binding of HCMV particles to fibroblasts, we could for the first time show that virion gH/gL/gO binds to platelet-derived growth factor-α (PDGFR-α) on the surface of fibroblasts and that gH/gL/gO either directly or indirectly recruits gB to this complex...
April 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/28400599/lps-promotes-a-monocyte-phenotype-permissive-for-human-cytomegalovirus-immediate-early-gene-expression-upon-infection-but-not-reactivation-from-latency
#10
V G Kew, M R Wills, M B Reeves
Human cytomegalovirus (HCMV) infection of myeloid cells is closely linked with the differentiation status of the cell. Haematopoietic progenitors and CD14+ monocytes are usually non-permissive for lytic gene expression which can lead to the establishment of latent infections. In contrast, differentiation to macrophage or dendritic cell (DC) phenotypes promotes viral reactivation or renders them permissive for lytic infection. The observation that high doses of Lipopolysaccharide (LPS) drove rapid monocyte differentiation in mice led us to investigate the response of human monocytes to HCMV following LPS stimulation in vitro...
April 11, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28394283/single-round-infectious-particle-antiviral-screening-assays-for-the-japanese-encephalitis-virus
#11
Chien-Yi Lu, Mann-Jen Hour, Ching-Ying Wang, Su-Hua Huang, Wen-Xiang Mu, Yu-Chun Chang, Cheng-Wen Lin
Japanese Encephalitis virus (JEV) is a mosquito-borne flavivirus with a positive-sense single-stranded RNA genome that contains a big open reading frame (ORF) flanked by 5'- and 3'- untranslated regions (UTRs). Nearly 30,000 JE cases with 10,000 deaths are still annually reported in East Asia. Although the JEV genotype III vaccine has been licensed, it elicits a lower protection against other genotypes. Moreover, no effective treatment for a JE case is developed. This study constructed a pBR322-based and cytomegaloviruses (CMV) promoter-driven JEV replicon for the production of JEV single-round infectious particles (SRIPs) in a packaging cell line expressing viral structural proteins...
April 10, 2017: Viruses
https://www.readbyqxmd.com/read/28393254/antitumor-effects-of-a-dual-specific-lentiviral-vector-carrying-the-escherichia-coli-purine-nucleoside-phosphorylase-gene
#12
Xiaolong Wang, Lei Sun, Xuejun Sun, Junhui Yu, Kai Wang, Yunhua Wu, Qi Gao, Jianbao Zheng
The Escherichia coli purine nucleoside phosphorylase/Fludarabine phosphate (ePNP/Fludara) suicide system has several drawbacks, such as side-effects and the low efficiency of ePNP expression. In this study, we evaluated the antitumor effects of the dual-specific 8HSEs-hTERTp-ePNP/Fludara suicide system under hyperthermia in vitro and in vivo. Luciferase activities from the 8HSEs‑hTERT and CMV promoters were compared using the dual luciferase assay in SW480 (high hTERT expression) and MKN74 cells (hTERT-negative) in the presence and absence of hyperthermia...
May 2017: International Journal of Oncology
https://www.readbyqxmd.com/read/28386736/an-efficient-rapid-system-for-assaying-hbx-mediated-transactivation
#13
Xiaoling Zhou, Haijun Shi, Shaozhe Yang, Pingnan Sun
OBJECTIVES: To develop a rapid and accurate assay system for screening inhibitors or enhancing agents targeting the transactivation capability of hepatitis B virus X protein (HBx) that activates cellular promoters in host cells to facilitate viral replication. RESULTS: We constructed a new GFP-based reporter system which was different from a luciferase-based reporter system. Firstly, a FLAG-tagged HBx gene was inserted into an expression plasmid, resulting in plasmid pHBx...
April 6, 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28381568/impact-of-antibodies-and-strain-polymorphisms-on-cytomegalovirus-entry-and-spread-in-fibroblasts-and-epithelial-cells
#14
Xiaohong Cui, Daniel C Freed, Dai Wang, Ping Qiu, Fengsheng Li, Tong-Ming Fu, Lawrence M Kauvar, Michael A McVoy
Cytomegalovirus (CMV) entry into fibroblasts differs from entry into epithelial cells. CMV also spreads cell-to-cell and can induce syncytia. To gain insights into these processes, 27 antibodies targeting epitopes in CMV virion glycoprotein complexes, including gB, gH/gL, and the pentamer, were evaluated for their effects on viral entry and spread. No antibodies inhibited CMV spread in fibroblasts, including those with potent neutralizing activity against fibroblast entry, while all antibodies that neutralized epithelial entry also inhibited spread in epithelial cells and a correlation existed between the potencies of these two activities...
April 5, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28357596/combined-sense-antisense-alu-elements-activate-the-egfp-reporter-gene-when-stable-transfection
#15
Zhihong Ma, Xianglong Kong, Shufeng Liu, Shuxian Yin, Yuehua Zhao, Chao Liu, Zhanjun Lv, Xiufang Wang
Alu elements in the human genome are present in more than one million copies, accounting for 10% of the genome. However, the biological functions of most Alu repeats are unknown. In this present study, we detected the effects of Alu elements on EGFP gene expression using a plasmid system to find the roles of Alu elements in human genome. We inserted 5'-4TMI-Alus-CMV promoter-4TMI-Alus (or antisense Alus)-3' sequences into the pEGFP-C1 vector to construct expression vectors. We altered the copy number of Alus, the orientation of the Alus, and the presence of an enhancer (4TMI) in the inserted 5'-4TMI-Alus-CMV promoter-4TMI-Alus (or antisense Alus)-3' sequences...
March 29, 2017: Molecular Genetics and Genomics: MGG
https://www.readbyqxmd.com/read/28356534/mutual-interplay-between-the-human-cytomegalovirus-terminase-subunits-pul51-pul56-and-pul89-promotes-terminase-complex-formation
#16
Sebastian Neuber, Karen Wagner, Thomas Goldne, Peter Lischka, Lars Steinbrueck, Martin Messerle, Eva Maria Borst
Human cytomegalovirus (HCMV) genome encapsidation requires several essential viral proteins, among them pUL56, pUL89 and the recently described pUL51, which constitute the viral terminase. To gain insight into terminase complex assembly, we investigated interactions between the individual subunits. For analysis in the viral context, HCMV bacterial artificial chromosomes carrying deletions in the open reading frames encoding the terminase proteins were used. These experiments were complemented by transient transfection assays with plasmids expressing the terminase components...
March 29, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28352540/promoter-transgene-and-cell-line-effects-in-the-transfection-of-mammalian-cells-using-pdmaema-based-nano-stars
#17
Alexander Raup, Valérie Jérôme, Ruth Freitag, Christopher V Synatschke, Axel H E Müller
Non-viral transfection protocols are typically optimized using standard cells and reporter proteins, potentially underestimating cellular or transgene effects. Here such effects were studied for two human (Jurkat, HEK-293) and two rodent (CHO-K1, L929) cell lines and three fluorescent reporter proteins. Expression of the enhanced green fluorescent protein (EGFP) was studied under the control of the human elongation factor 1 alpha promoter and three viral promoters (SV40, SV40/enhancer, CMV), that of ZsYellow1 (yellow fluorescence) and mCherry (red fluorescence) for the CMV promoter...
September 2016: Biotechnology Reports
https://www.readbyqxmd.com/read/28350327/the-envelope-gene-of-hepatitis-b-virus-is-implicated-in-both-differential-virion-secretion-and-genome-replication-capacities-between-genotype-b-and-genotype-c-isolates
#18
Haodi Jia, Yanli Qin, Chaoyang Chen, Fei Zhang, Cheng Li, Li Zong, Yongxiang Wang, Jiming Zhang, Jisu Li, Yumei Wen, Shuping Tong
Chronic infection by hepatitis B virus (HBV) genotype C is associated with a prolonged replicative phase and an increased risk of liver cancer, compared with genotype B infection. We previously found lower replication capacity but more efficient virion secretion by genotype C than genotype B isolates. Virion secretion requires interaction between core particles and ENVELOPE proteins. In the present study, chimeric constructs between genotype B and genotype C clones were generated to identify the structural basis for differential virion secretion...
March 28, 2017: Viruses
https://www.readbyqxmd.com/read/28346434/optimization-of-adeno-associated-virus-vector-mediated-gene-transfer-to-the-respiratory-tract
#19
F Kurosaki, R Uchibori, N Mato, Y Sehara, Y Saga, M Urabe, H Mizukami, Y Sugiyama, A Kume
An efficient adeno-associated virus (AAV) vector was constructed for the treatment of respiratory diseases. AAV serotypes, promoters, and routes of administration potentially influencing the efficiency of gene transfer to airway cells were examined in the present study. Among the nine AAV serotypes (AAV1-9) screened in vitro and four serotypes (AAV1, 2, 6, 9) evaluated in vivo, AAV6 showed the strongest transgene expression. As for promoters, the cytomegalovirus (CMV) early enhancer/chicken β-actin (CAG) promoter resulted in more robust transduction than the CMV promoter...
March 27, 2017: Gene Therapy
https://www.readbyqxmd.com/read/28344994/a-new-broad-range-plasmid-for-dna-delivery-in-eukaryotic-cells-using-lactic-acid-bacteria-in%C3%A2-vitro-and-in%C3%A2-vivo-assays
#20
Pamela Mancha-Agresti, Mariana Martins Drumond, Fillipe Luiz Rosa do Carmo, Monica Morais Santos, Janete Soares Coelho Dos Santos, Franco Venanzi, Jean-Marc Chatel, Sophie Yvette Leclercq, Vasco Azevedo
Lactococcus lactis is well documented as a promising candidate for development of novel oral live vaccines. It has been broadly engineered for heterologous expression, as well as for plasmid expression vector delivery, directly inside eukaryotic cells, for DNA vaccine, or as therapeutic vehicle. This work describes the characteristics of a new plasmid, pExu (extra chromosomal unit), for DNA delivery using L. lactis and evaluates its functionality both by in vitro and in vivo assays. This plasmid exhibits the following features: (1) a theta origin of replication and (2) an expression cassette containing a multiple cloning site and a eukaryotic promoter, the cytomegalovirus (pCMV)...
March 17, 2017: Molecular Therapy. Methods & Clinical Development
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