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https://www.readbyqxmd.com/read/28548037/moonlighting-glycolytic-enzyme-glyceraldehyde-3-phosphate-dehydrogenase-gapdh-is-required-for-efficient-hepatitis-c-virus-and-dengue-virus-infections-in-human-huh-7-5-1-cells
#1
Meera Raj, Mary Langley, Steven J McArthur, François Jean
Hijacking of cellular biosynthetic pathways by human enveloped viruses is a shared molecular event essential for the viral lifecycle. In this study, the accumulating evidence of the importance of human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in the host secretory pathway led us to hypothesize that this moonlighting enzyme could play a key role in the lifecycle steps of two important Flaviviridae members, hepatitis C virus (HCV) and dengue virus (DENV). We used short interfering RNA (siRNA)-mediated knockdown of human GAPDH in Huh-7...
May 26, 2017: Journal of General Virology
https://www.readbyqxmd.com/read/28539455/functional-consequences-of-rna-5-terminal-deletions-on-coxsackievirus-b3-rna-replication-and-ribonucleoprotein-complex-formation
#2
Nicolas Lévêque, Magali Garcia, Alexis Bouin, Joseph H C Nguyen, Genevieve P Tran, Laurent Andreoletti, Bert L Semler
Group B coxsackieviruses are responsible for chronic cardiac infections. However, the molecular mechanisms by which the virus can persist in the human heart long after the signs of acute myocarditis have abated are still not completely understood. Recently, coxsackievirus B3 strains with 5' terminal deletions in genomic RNAs were isolated from a patient suffering from idiopathic dilated cardiomyopathy, suggesting that such mutant viruses may be the forms responsible for persistent infection. These deletions lacked portions of 5' stem-loop I, which is an RNA secondary structure required for viral RNA replication...
May 24, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28535790/clonal-replacement-of-epidemic-kpc-producing-klebsiella-pneumoniae-in-a-hospital-in-china
#3
Yuying Liang, Xiuyun Yin, Lijun Zeng, Shuiping Chen
BACKGROUND: Klebsiella pneumoniae is a frequent nosocomial pathogen causing difficult-to-treat infections worldwide. The prevalence of Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae (KPC-KP) is increasing in China. The aim of this study was to investigate the molecular epidemiology of KPC-KP in a nosocomial outbreak. METHODS: Fifty-four KPC-KP isolates were consecutively collected between November 2013 and August 2014 during a KPC-KP outbreak in a tertiary care hospital in Beijing, China...
May 23, 2017: BMC Infectious Diseases
https://www.readbyqxmd.com/read/28522548/a-multi-purpose-toolkit-to-enable-advanced-genome-engineering-in-plants
#4
Tomas Cermak, Shaun J Curtin, Javier Gil-Humanes, Radim Čegan, Thomas J Y Kono, Eva Konečná, Joseph J Belanto, Colby G Starker, Jade W Mathre, Rebecca L Greenstein, Daniel F Voytas
We report a comprehensive toolkit that enables targeted, specific modification of monocot and dicot genomes using a variety of genome engineering approaches. Our reagents, based on TALENs and the CRISPR/Cas9 system, are systematized for fast, modular cloning and accommodate diverse regulatory sequences to drive reagent expression. Vectors are optimized to create either single or multiple gene knockouts and large chromosomal deletions. Moreover, integration of geminivirus-based vectors enables precise gene editing through homologous recombination...
May 18, 2017: Plant Cell
https://www.readbyqxmd.com/read/28516137/a-curated-dataset-of-complete-enterobacteriaceae-plasmids-compiled-from-the-ncbi-nucleotide-database
#5
Alex Orlek, Hang Phan, Anna E Sheppard, Michel Doumith, Matthew Ellington, Tim Peto, Derrick Crook, A Sarah Walker, Neil Woodford, Muna F Anjum, Nicole Stoesser
Thousands of plasmid sequences are now publicly available in the NCBI nucleotide database, but they are not reliably annotated to distinguish complete plasmids from plasmid fragments, such as gene or contig sequences; therefore, retrieving complete plasmids for downstream analyses is challenging. Here we present a curated dataset of complete bacterial plasmids from the clinically relevant Enterobacteriaceae family. The dataset was compiled from the NCBI nucleotide database using curation steps designed to exclude incomplete plasmid sequences, and chromosomal sequences misannotated as plasmids...
June 2017: Data in Brief
https://www.readbyqxmd.com/read/28508218/recovery-of-a-paramyxovirus-the-human-metapneumovirus-from-cloned-cdna
#6
B G van den Hoogen, R A M Fouchier
Human metapneumovirus (HMPV), a single-stranded negative-sense RNA virus belonging to the family Paramyxoviridae, is associated with respiratory tract illness, primarily in young children and persons with underlying disease. Based on genetic and antigenic variation, HMPV strains are classified into two serotypes, with isolates NL/1/00 and NL/1/99 as prototypes for serotypes A and B, respectively. The development of plasmid-based reverse genetics systems for both serotypes has resulted in developments of a wide range of vaccine candidates against HMPV infection...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28466040/agroinfection-of-tobacco-by-croton-yellow-vein-mosaic-virus-and-designing-of-a-replicon-vector-for-expression-of-foreign-gene-in-plant
#7
A Abdul Kader Jailani, Alok Kumar, Bikash Mandal, T Sivasudha, Anirban Roy
Croton yellow vein mosaic virus (CYVMV, genus Begomovirus family Geminiviridae) is a proliferating begomovirus in the Indian sub-continent. The infectious constructs in binary vector was developed against the CYVMV genome and its associated betasatellite. Agroinoculation of the genomic construct of CYVMV produced leaf curl symptoms alone in three species of tobacco, Nicotiana tabacum, N. benthamiana and N. glutinosa. Co-inoculation of betasatellite enhanced the severity of the disease and reduced the incubation time...
September 2016: Virusdisease
https://www.readbyqxmd.com/read/28458051/emergence-and-nosocomial-spread-of-carbapenem-resistant-oxa-232-producing-klebsiella-pneumoniae-in-brunei-darussalam
#8
Muhd Haziq Fikry Abdul Momin, Apostolos Liakopoulos, Lynette M Phee, David W Wareham
OBJECTIVES: Carbapenem-resistant Enterobacteriaceae (CRE) are identified as a major global health concern. The success of CRE is facilitated by the emergence, acquisition and spread of successful clones carrying plasmid-encoded resistance genes. In this study, an outbreak of carbapenem-resistant Klebsiella pneumoniae (CRKP) infections in patients hospitalised in Brunei Darussalam was investigated. METHODS: Over a 3-month period (May-July 2015), five multidrug-resistant K...
April 27, 2017: Journal of Global Antimicrobial Resistance
https://www.readbyqxmd.com/read/28456632/tumor-necrosis-factor-inhibits-spread-of-hepatitis-c-virus-among-liver-cells-independent-from-interferons
#9
Stephen M Laidlaw, Svetlana Marukian, Rachel H Gilmore, Siobhán B Cashman, Volodymyr Nechyporuk-Zloy, Charles M Rice, Lynn B Dustin
BACKGROUND & AIMS: Tumor necrosis factor (TNF) an inflammatory cytokine expressed by human fetal liver cells (HFLCs) following infection with cell culture-derived hepatitis C virus. TNF has been reported to increase entry of HCV pseudoparticles into hepatoma cells and inhibit signaling by interferon alpha (IFNA), but have no effect on replication of HCV RNA. We investigated the effects of TNF on HCV infection of and spread among Huh-7 hepatoma cells and primary HFLCs.  METHODS: Human hepatoma (Huh-7 and Huh-7...
April 26, 2017: Gastroenterology
https://www.readbyqxmd.com/read/28456022/cellular-dead-box-rna-helicase-ddx6-modulates-interaction-of-mir-122-with-the-5-untranslated-region-of-hepatitis-c-virus-rna
#10
Jason M Biegel, Eric Henderson, Erica M Cox, Gaston Bonenfant, Rachel Netzband, Samantha Kahn, Rachel Eager, Cara T Pager
Hepatitis C virus (HCV) subverts the cellular DEAD-box RNA helicase DDX6 to promote virus infection. Using polysome gradient analysis and the subgenomic HCV Renilla reporter replicon genome, we determined that DDX6 does not affect HCV translation. Rather expression of the subgenomic HCV Renilla luciferase reporter at late times, as well as labeling of newly synthesized viral RNA with 4-thiouridine showed that DDX6 modulates replication. Because DDX6 is an effector protein of the microRNA pathway, we also investigated its role in miR-122-directed HCV gene expression...
April 26, 2017: Virology
https://www.readbyqxmd.com/read/28446899/clonal-spread-of-16s-rrna-methyltransferase-producing-klebsiella-pneumoniae-st37-with-high-prevalence-of-esbls-from-companion-animals-in-china
#11
Jing Xia, Liang-Xing Fang, Ke Cheng, Guo-Hao Xu, Xi-Ran Wang, Xiao-Ping Liao, Ya-Hong Liu, Jian Sun
We screened 30 Klebsiella pneumoniae isolates from dogs and cats at a single animal hospital in Guangdong Province, China. Among them, 12 K. pneumoniae strains possessed high-level resistance to amikacin and gentamicin and these were screened for 16S rRNA methyltransferase (16S-RMTase) genes. And then the genes positive isolates were detected for ESBLs (extended spectrum β-lactamases) and analyzed by pulsed-field gel electrophoresis, multilocus sequence typing, PCR-based replicon typing and plasmid analysis...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28443083/intracellular-localization-interactions-and-functions-of-capsicum-chlorosis-virus-proteins
#12
Shirani M K Widana Gamage, Ralf G Dietzgen
Tospoviruses are among the most devastating viruses of horticultural and field crops. Capsicum chlorosis virus (CaCV) has emerged as an important pathogen of capsicum and tomato in Australia and South-east Asia. Present knowledge about CaCV protein functions in host cells is lacking. We determined intracellular localization and interactions of CaCV proteins by live plant cell imaging to gain insight into the associations of viral proteins during infection. Proteins were transiently expressed as fusions to autofluorescent proteins in leaf epidermal cells of Nicotiana benthamiana and capsicum...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28443075/grim-19-restricts-hcv-replication-by-attenuating-intracellular-lipid-accumulation
#13
Jung-Hee Kim, Pil S Sung, Eun B Lee, Wonhee Hur, Dong J Park, Eui-Cheol Shin, Marc P Windisch, Seung K Yoon
Gene-associated with retinoid-interferon-induced mortality 19 (GRIM-19) targets multiple signaling pathways involved in cell death and growth. However, the role of GRIM-19 in the pathogenesis of hepatitis virus infections remains unexplored. Here, we investigated the restrictive effects of GRIM-19 on the replication of hepatitis C virus (HCV). We found that GRIM-19 protein levels were reduced in HCV-infected Huh7 cells and Huh7 cells harboring HCV replicons. Moreover, ectopically expressed GRIM-19 caused a reduction in both intracellular viral RNA levels and secreted viruses in HCVcc-infected cell cultures...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28441762/replication-and-oncolytic-activity-of-an-avian-orthoreovirus-in-human-hepatocellular-carcinoma-cells
#14
Robert A Kozak, Larissa Hattin, Mia J Biondi, Juan C Corredor, Scott Walsh, Max Xue-Zhong, Justin Manuel, Ian D McGilvray, Jason Morgenstern, Evan Lusty, Vera Cherepanov, Betty-Anne McBey, David Leishman, Jordan J Feld, Byram Bridle, Éva Nagy
Oncolytic viruses are cancer therapeutics with promising outcomes in pre-clinical and clinical settings. Animal viruses have the possibility to avoid pre-existing immunity in humans, while being safe and immunostimulatory. We isolated an avian orthoreovirus (ARV-PB1), and tested it against a panel of hepatocellular carcinoma cells. We found that ARV-PB1 replicated well and induced strong cytopathic effects. It was determined that one mechanism of cell death was through syncytia formation, resulting in apoptosis and induction of interferon stimulated genes (ISGs)...
April 24, 2017: Viruses
https://www.readbyqxmd.com/read/28439036/reclassification-of-the-specialized-metabolite-producer-pseudomonas-mesoacidophila-atcc-31433-as-a-member-of-the-burkholderia-cepacia-complex
#15
E Joel Loveridge, Cerith Jones, Matthew J Bull, Suzy C Moody, Małgorzata W Kahl, Zainab Khan, Louis Neilson, Marina Tomeva, Sarah E Adams, Andrew C Wood, Daniel Rodriguez-Martin, Ingrid Pinel, Julian Parkhill, Eshwar Mahenthiralingam, John Crosby
Pseudomonas mesoacidophila ATCC 31433 is a Gram-negative bacterium, first isolated from Japanese soil samples, which produces the monobactam isosulfazecin and the β-lactam potentiating bulgecins. To characterize the biosynthetic potential of P. mesoacidophila ATCC 31433 its complete genome was determined using single molecule real time DNA sequence analysis. The 7.8 Mb genome comprised four replicons: three chromosomal, each encoding ribosomal RNA, and one plasmid. Phylogenetic analysis demonstrated that P...
April 24, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/28438609/novel-replicons-and-trans-encapsidation-systems-for-hepatitis-c-virus-proteins-live-imaging-and-virus-host-interaction-proteomics
#16
Ovidiu Vlaicu, Tudor Selescu, Florin Pastrama, Cristian Munteanu, Laura Riva, Jean Dubuisson, Yves Rouille, Costin-Ioan Popescu
Proteomics and imaging techniques are used more and more in tandem to investigate the virus-host interaction. Herein we present novel replicons, methods and trans-encapsidation systems suitable for determination of Hepatitis C Virus (HCV) proteins interactomes and live imaging of viral proteins dynamics in HCV cell culture (HCVcc) system. To identify endogenous factors involved in the HCV life cycle, we constructed full-length functional replicons with affinity purification (AP) tags fused to NS2 and NS5A proteins...
April 21, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28435072/development-of-a-pseudovirus-based-assay-for-measuring-neutralizing-antibodies-against-coxsackievirus-b5
#17
Pan Chen, Xing Wu, Yao Su, Xiaotian Hao, Qunying Mao, Zhenglun Liang
Coxsackievirus B5 (CV-B5), an important Coxsackie B virus from genus Enteroviruse within the family Picornaviridae, has also been isolated from Hand, Foot, and Mouth Disease (HFMD) patients, and often associated with neurological manifestations. In this study, we found out that Coxsackievirus B3 (CV-B3) replicon RNA could be encapsidated with CV-B5 capsid to assemble infectious CV-B5 pseudovirus. We then utilized this single round infection system of CV-B5 to develop a neutralizing antibody quantification assay...
April 20, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28432094/the-use-of-synthetic-hybrid-strains-to-determine-the-role-of-replicon-3-in-virulence-of-the-burkholderia-cepacia-complex
#18
Kirsty Agnoli, Roman Freitag, Margarida C Gomes, Christian Jenul, Angela Suppiger, Olga Mannweiler, Carmen Frauenknecht, Daniel Janser, Annette C Vergunst, Leo Eberl
The Burkholderia cepacia complex (Bcc) displays a wealth of metabolic diversity with great biotechnological potential, but the utilisation of these bacteria is limited by their opportunistic pathogenicity to humans. The third replicon of the Bcc, megaplasmid pC3 (0.5 to 1.4 Mb, previously chromosome 3), is important for various phenotypes, including virulence, antifungal and proteolytic activities, and the utilization of certain substrates. Approximately half of plasmid pC3 is well-conserved throughout sequenced Bcc members, while the other half is not...
April 21, 2017: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/28430437/discovery-of-a-hepatitis-c-virus-ns5b-replicase-palm-site-allosteric-inhibitor-bms-929075-advanced-to-phase-1-clinical-studies
#19
Kap-Sun Yeung, Brett R Beno, Kyle Parcella, John A Bender, Katherine A Grant-Young, Andrew Nickel, Prashantha Gunaga, Prakash Anjanappa, Rajesh Onkardas Bora, Kumaravel Selvakumar, Karen Rigat, Ying-Kai Wang, Mengping Liu, Julie Lemm, Kathy Mosure, Steven Sheriff, Changhong Wan, Mark Witmer, Kevin Kish, Umesh Hanumegowda, Xiaoliang Zhuo, Yue-Zhong Shu, Dawn Parker, Roy Haskell, Alicia Ng, Qi Gao, Elizabeth Colston, Joseph Raybon, Dennis M Grasela, Kenneth Santone, Min Gao, Nicholas A Meanwell, Michael Sinz, Matthew G Soars, Jay O Knipe, Susan B Roberts, John F Kadow
The hepatitis C virus (HCV) NS5B replicase is a prime target for the development of direct-acting antiviral drugs for the treatment of chronic HCV infection. Inspired by the overlay of bound structures of three structurally distinct NS5B palm site allosteric inhibitors, the high-throughput screening hit anthranilic acid 4, the known benzofuran analogue 5, and the benzothiadiazine derivative 6, an optimization process utilizing the simple benzofuran template 7 as a starting point for a fragment growing approach was pursued...
May 4, 2017: Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28427827/antiviral-screen-identifies-ev71-inhibitors-and-reveals-camptothecin-target-dna-topoisomerase-1-as-a-novel-ev71-host-factor
#20
Kan Xing Wu, Justin Jang-Hann Chu
Enterovirus 71 (EV71) is one of the causative agents of hand, foot and mouth disease (HFMD) associated with severe neurological disease. EV71's pathogenesis remains poorly understood and the lack of approved antiviral has led to its emergence as a clinically important neurotropic virus. The goals of this study were to: (i) identify novel anti-EV71 compounds that may serve as lead molecules for therapeutics; and (ii) investigate their targets in downstream studies. We screened a 502-compound library of highly purified natural products for anti-EV71 activities in a cell-based immunofluorescence assay that were then confirmed in viral plaque reduction assays...
April 17, 2017: Antiviral Research
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