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https://www.readbyqxmd.com/read/28645197/pht%C3%AE-promoted-mobilization-of-non-conjugative-resistance-plasmids-from-enterococcus-faecium-to-enterococcus-faecalis
#1
Laura Di Sante, Gianluca Morroni, Andrea Brenciani, Carla Vignaroli, Alberto Antonelli, Marco Maria D'Andrea, Andrea Di Cesare, Eleonora Giovanetti, Pietro E Varaldo, Gian Maria Rossolini, Francesca Biavasco
Objectives: : To analyse the recombination events associated with conjugal mobilization of two multiresistance plasmids, pRUM 17i48 and pLAG (formerly named pDO1-like), from Enterococcus faecium 17i48 to Enterococcus faecalis JH2-2. Methods: : The plasmids from two E. faecalis transconjugants (JH-4T, tetracycline resistant, and JH-8E, erythromycin resistant) and from the E. faecium donor (also carrying a pHTβ-like conjugative plasmid, named pHTβ 17i48 ) were investigated by several methods, including PCR mapping and sequencing, S1-PFGE followed by Southern blotting and hybridization, and WGS...
June 22, 2017: Journal of Antimicrobial Chemotherapy
https://www.readbyqxmd.com/read/28645187/salmonella-virulence-plasmid-pathogenesis-and-ecology
#2
Claudia Silva, José Luis Puente, Edmundo Calva
A current view on the role of the Salmonella virulence plasmid in the pathogenesis of animal and human hosts is discussed; including the possible relevance in secondary ecological niches. Various strategies towards further studies in this respect are proposed within the One Health Concept.
June 22, 2017: Pathogens and Disease
https://www.readbyqxmd.com/read/28645177/mobile-genetic-elements-in-neisseria-gonorrhoeae-movement-for-change
#3
Ana Cehovin, Steven B Lewis
Neisseria gonorrhoeae, the causative agent of the sexually transmitted disease gonorrhoeae, possesses several mobile genetic elements (MGEs). The MGEs such as transposable elements mediate intra-chromosomal rearrangements, while plasmids and the gonococcal genetic island are involved in inter-chromosomal gene transfer. Additionally, gonococcal MGEs serve as hotspots for recombination and integration of other genetic elements such as bacteriophages, contribute to gene regulation or spread genes through gonococcal populations by horizontal gene transfer...
June 22, 2017: Pathogens and Disease
https://www.readbyqxmd.com/read/28644573/characteristics-of-integrons-and-associated-gene-cassettes-in-antibiotic-resistant-escherichia-coli-isolated-from-free-ranging-food-animals-in-china
#4
Mujeeb Ur Rehman, Hui Zhang, Shucheng Huang, Muhammad Kashif Iqbal, Khalid Mehmood, Houqiang Luo, Jiakui Li
We investigated the occurrence of integrons in antibiotic-resistant Escherichia coli strains isolated from free-ranging food animals, including yaks, piglets, and chickens, in China, and characterized the gene cassettes harbored within the integrons. We examined 432 E. coli strains that exhibited resistance to at least one class of antibiotics. Integrase genes and associated gene cassettes were characterized by polymerase chain reaction (PCR) analysis, restriction fragment-length polymorphism, DNA sequencing, conjugation experiments, and plasmid analysis...
June 23, 2017: Journal of Food Science
https://www.readbyqxmd.com/read/28644560/identification-and-characterization-of-ars-like-sequences-as-putative-origin-s-of-replication-in-human-malaria-parasite-plasmodium-falciparum
#5
Meetu Agarwal, Krishanu Bhowmick, Kushal Shah, Annangarachari Krishnamachari, Suman Kumar Dhar
DNA replication is a fundamental process in genome maintenance, and initiates from several genomic sites (origins) in eukaryotes. In S. cerevisiae, conserved sequences known as Autonomously Replicating Sequences (ARSs) provide a landing pad for the Origin Recognition Complex (ORC), leading to replication initiation. Although origins from higher eukaryotes share some common sequence features, the definitive genomic organization of these sites remains elusive. The human malaria parasite Plasmodium falciparum undergoes multiple rounds of DNA replication; therefore, control of initiation events is crucial to ensure proper replication...
June 23, 2017: FEBS Journal
https://www.readbyqxmd.com/read/28643952/asn194lys-mutation-in-rvg29-peptide-increases-gfp-transgene-delivery-by-endocytosis-to-neuroblastoma-and-astrocyte-cells
#6
Sheila Adela Villa-Cedillo, Humberto Rodríguez-Rocha, Laura Mireya Zavala-Flores, Roberto Montes-de-Oca-Luna, Aracely García-García, Maria de Jesus Loera-Arias, Odila Saucedo-Cárdenas
OBJECTIVES: A cell-penetrating peptide-based delivery system could target specific types of cells for therapeutic genes delivery. To increase the gene delivery efficiency into neuronal phenotype cells, we introduced an Asn194Lys mutation to RVG29 peptide derived from rabies virus glycoprotein and added a nuclear localization signal to enhance its nuclear import. METHODS: Mutant RVG or wild-type RVG peptide, a karyophilic peptide (KP) and a plasmid encoding green fluorescent protein (pGL) were bound by electrostatic charges to form four different kinds of RVG complexes...
June 23, 2017: Journal of Pharmacy and Pharmacology
https://www.readbyqxmd.com/read/28643761/the-effects-of-gene-therapy-with-granulocyte-macrophage-colony-stimulating-factor-in-the-regression-of-tumor-masses-in-fibrosarcoma-mouse-model
#7
Saiedeh RaziSoofiyani, Tohid Kazemi, Farzaneh Lotfipour, Leila Mohammadnejad, Somayeh Hallaj-Nezhadi, Siamak Sandoghchian Shotorbani, Akbar Mohammad Hosseini, Behzad Baradaran
INTRODUCTION: Cytokine gene therapy is one of the cancer treatment strategies. Recently, granulocyte-monocyte colony-stimulating factor (GM-CSF), as an important cytokine in activating dendritic cells and boosting the anti-tumor immune responses, has been utilized as an immunotherapeutic agent in cancer gene therapy. The purpose of the present investigation was to study the GM-CSF gene therapy effects in regression of tumor masses in fibrosarcoma mouse model. MATERIALS AND METHODS: To investigate the therapeutic efficacy of GM-CSF, WEHI-164 tumor cells were transfected with murine GM-CSF plasmid...
April 2017: Journal of Cancer Research and Therapeutics
https://www.readbyqxmd.com/read/28643490/characterization-of-extended-spectrum-%C3%AE-lactamase-genes-of-shigella-flexneri-isolates-with-fosfomycin-resistance-from-patients-in-china
#8
Yanyan Liu, Yue Cheng, Haifei Yang, Lifen Hu, Jun Cheng, Ying Ye, Jiabin Li
BACKGROUND: The emergence of fosfomycin resistance and extended-spectrum β-lactamase (ESBL) genes is a serious threat to public health and a new challenge in shigellosis treatment. The purpose of this study was to identify fosfomycin resistance and characterize β-lactamase genes in fos-carrying isolates of Shigella flexneri from patients in China. METHODS: A total of 263 S. flexneri isolates were collected from 34 hospitals in the Anhui Province of China during September 2012-September 2015 and screened for fosA3, fosA, and fosC2 by PCR amplification and sequencing...
September 2017: Annals of Laboratory Medicine
https://www.readbyqxmd.com/read/28643488/multidrug-resistance-mechanisms-of-carbapenem-resistant-klebsiella-pneumoniae-strains-isolated-in-chongqing-china
#9
Jinrong Yan, Shuli Pu, Xiaojiong Jia, Xiuyu Xu, Shuangshuang Yang, Jing Shi, Shan Sun, Liping Zhang
BACKGROUND: Carbapenem-resistant Klebsiella pneumoniae (CRKP) is considered a serious global threat. However, little is known regarding the multidrug resistance (MDR) mechanisms of CRKP. This study investigated the phenotypes and MDR mechanisms of CRKP and identified their clonal characteristics. METHODS: PCR and sequencing were utilized to identify antibiotic resistance determinants. Integron gene cassette arrays were determined by restriction fragment length polymorphism (RFLP) analysis...
September 2017: Annals of Laboratory Medicine
https://www.readbyqxmd.com/read/28643259/genome-editing-of-medaka
#10
Satoshi Ansai, Masato Kinoshita
Medaka (Oryzias latipes), along with zebrafish (Danio rerio), is a useful experimental model fish. Here, we describe a simple method for generating medaka gene knock-out strains using an automated microchip electrophoresis system. We also describe a method for targeted gene knock-in using a plasmid carrying a sequence that does not cause off-target effects in medaka.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28643254/gene-targeting-in-rabbits-single-step-generation-of-knock-out-rabbits-by-microinjection-of-crispr-cas9-plasmids
#11
Yoshihiro Kawano, Arata Honda
The development of genome editing technology has allowed gene disruptions to be achieved in various animal species and has been beneficial to many mammals. Gene disruption using pluripotent stem cells is difficult to achieve in rabbits, but thanks to advances in genome editing technology, a number of gene disruptions have been conducted. This paper describes a simple and easy method for carrying out gene disruptions in rabbits using CRISPR/Cas9 in which the gene to be disrupted is marked, the presence or absence of off-target candidates is checked, and a plasmid allowing simultaneous expression of Cas9 and sgRNA is constructed...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28643250/genome-editing-in-mouse-zygotes-and-embryonic-stem-cells-by-introducing-sgrna-cas9-expressing-plasmids
#12
Taichi Noda, Asami Oji, Masahito Ikawa
In mammalian cells, genome editing with the single guide RNA (sgRNA)/Cas9 complex allows for high targeting efficiency within a relatively short time frame with the added benefits of being low cost and easy to design. sgRNA/Cas9-mediated editing in mouse zygotes has accelerated the analysis of gene functions and the generation of mouse models of human diseases. Despite the benefits, this method still suffers from several problems, such as mosaicism in the founder generation which complicates genotyping and phenotypical analyses, and the low efficiency of more complicated genome editing...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28642992/plasmid-mediated-quinolone-resistance-pmqr-genes-and-class-1-integrons-in-quinolone-resistant-marine-bacteria-and-clinical-isolates-of-escherichia-coli-from-an-aquacultural-area
#13
Alexandra Tomova, Larisa Ivanova, Alejandro H Buschmann, Henry P Godfrey, Felipe C Cabello
Antimicrobial usage in aquaculture selects for antimicrobial-resistant microorganisms in the marine environment. The relevance of this selection to terrestrial animal and human health is unclear. Quinolone-resistance genes qnrA, qnrB, and qnrS were chromosomally located in four randomly chosen quinolone-resistant marine bacteria isolated from an aquacultural area with heavy quinolone usage. In quinolone-resistant uropathogenic clinical isolates of Escherichia coli from a coastal area bordering the same aquacultural region, qnrA was chromosomally located in two E...
June 23, 2017: Microbial Ecology
https://www.readbyqxmd.com/read/28642849/aim2-co-immunization-with-vp1-is-associated-with-increased-memory-cd8-t-cells-and-mounts-long-lasting-protection-against-coxsackievirus-b3-challenge
#14
Liang Yin, Dafei Chai, Yan Yue, Chunsheng Dong, Sidong Xiong
The recurrent Coxsackievirus B3 (CVB3) infection is the most important cause of intractable myocarditis which often leads to chronic myocarditis and even dilated cardiomyopathy. Therefore, enhanced DNA vaccines capable of memory CD8 T cells are essential for long-lasting immunological protection against CVB3 infection. In this study, absent in melanoma 2 (AIM2) was used as an adjuvant to enhance the induction of memory CD8 T cells elicited by VP1 (viral capsid protein 1) vaccine. Mice were intramuscularly injected with 50 μg AIM2 plasmid and equal amount of VP1 plasmid (pAIM2/pVP1) vaccine 4 times at 2 week-intervals...
2017: Frontiers in Cellular and Infection Microbiology
https://www.readbyqxmd.com/read/28642846/decreased-fitness-and-virulence-in-st10-escherichia-coli-harboring-blandm-5-and-mcr-1-against-a-st4981-strain-with-blandm-5
#15
Yawei Zhang, Kang Liao, Hua Gao, Qi Wang, Xiaojuan Wang, Henan Li, Ruobing Wang, Hui Wang
Although coexistence of blaNDM-5 and mcr-1 in Escherichia coli has been reported, little is known about the fitness and virulence of such strains. Three carbapenem-resistant Escherichia coli (GZ1, GZ2, and GZ3) successively isolated from one patient in 2015 were investigated for microbiological fitness and virulence. GZ1 and GZ2 were also resistant to colistin. To verify the association between plasmids and fitness, growth kinetics of the transconjugants were performed. We also analyzed genomic sequences of GZ2 and GZ3 using PacBio sequencing...
2017: Frontiers in Cellular and Infection Microbiology
https://www.readbyqxmd.com/read/28642136/a-novel-hepatitis-b-virus-derived-cis-acting-sequence-that-enhances-expression-of-transgenes-delivered-by-plasmid-vectors-in-eukaryote-cell-culture-systems
#16
Seoung-Ae Lee, Hong Kim, So-Young Lee, Bum-Joon Kim
We tested the effectiveness of a novel 13-bp hepatitis B virus (HBV)-derived cis-acting element (CAE) (ACCTCGACAAGGC), called the DT2 CAE, in augmenting transgene expression delivered by plasmid vectors in eukaryotic cells. The addition of the DT2 CAE just upstream of the start codon of several different target proteins (luciferase, EGFP, LHB, HBsAg, and MIF) in DNA plasmid constructs enhanced their translation in a posttranscriptional manner, irrespective of cell type (cell lines or primary cells) or promoter (CMV or HBV preS1 promoters), suggesting its feasibility for enhanced protein production in eukaryotic cell systems...
June 19, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28641931/transfer-of-antibiotic-resistance-in-staphylococcus-aureus
#17
REVIEW
Jakob Haaber, José R Penadés, Hanne Ingmer
Staphylococcus aureus is a serious human pathogen with remarkable adaptive powers. Antibiotic-resistant clones rapidly emerge mainly by acquisition of antibiotic-resistance genes from other S. aureus strains or even from other genera. Transfer is mediated by a diverse complement of mobile genetic elements and occurs primarily by conjugation or bacteriophage transduction, with the latter traditionally being perceived as the primary route. Recent work on conjugation and transduction suggests that transfer by these mechanisms may be more extensive than previously thought, in terms of the range of plasmids that can be transferred by conjugation and the efficiency with which transduction occurs...
June 19, 2017: Trends in Microbiology
https://www.readbyqxmd.com/read/28640821/noise-reduction-in-single-time-frame-optical-dna-maps
#18
Paola C Torche, Vilhelm Müller, Fredrik Westerlund, Tobias Ambjörnsson
In optical DNA mapping technologies sequence-specific intensity variations (DNA barcodes) along stretched and stained DNA molecules are produced. These "fingerprints" of the underlying DNA sequence have a resolution of the order one kilobasepairs and the stretching of the DNA molecules are performed by surface adsorption or nano-channel setups. A post-processing challenge for nano-channel based methods, due to local and global random movement of the DNA molecule during imaging, is how to align different time frames in order to produce reproducible time-averaged DNA barcodes...
2017: PloS One
https://www.readbyqxmd.com/read/28640659/phlda1-pleckstrin-homology-like-domain-family-a-member-1-knockdown-promotes-migration-and-invasion-of-mcf10a-breast-epithelial-cells
#19
Naieli Bonatto, Maria José Carlini, Simone Aparecida de Bessa Garcia, Maria Aparecida Nagai
PHLDA1 (pleckstrin homology-like domain, family A, member 1) is a multifunctional protein that plays distinct roles in several biological processes including cell death and therefore its altered expression has been identified in different types of cancer. Progressively loss of PHLDA1 was found in primary and metastatic melanoma while its overexpression was reported in intestinal and pancreatic tumors. Previous work from our group showed that negative expression of PHLDA1 protein was a strong predictor of poor prognosis for breast cancer disease...
June 22, 2017: Cell Adhesion & Migration
https://www.readbyqxmd.com/read/28640456/first-environmental-sample-containing-plasmid-mediated-colistin-resistant-esbl-producing-escherichia-coli-detected-in-norway
#20
Silje Bakken Jørgensen, Arne Søraas, Lotte Stenfors Arnesen, Truls Leegaard, Arnfinn Sundsfjord, Pål A Jenum
We hereby report a detection of the plasmid borne mcr-1 gene conferring colistin resistance in an extended-spectrum β-lactamase (ESBL) producing Escherichia coli ST10 strain retrieved from seawater at a public beach in Norway. The sample was collected in September 2010 and was investigated by whole-genome sequencing in 2016. This report illustrates that E. coli strains carrying plasmid-mediated colistin resistance genes have also reached areas where this drug is hardly used at all. Surveillance of colistin resistance in environmental, veterinary, and human strains is warranted also in countries where colistin resistance is rare in clinical settings...
June 22, 2017: APMIS: Acta Pathologica, Microbiologica, et Immunologica Scandinavica
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