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https://www.readbyqxmd.com/read/28350472/development-of-a-cho-based-cell-free-platform-for-synthesis-of-active-monoclonal-antibodies
#1
Rey W Martin, Natalia I Majewska, Cindy X Chen, Thomas E Albanetti, Rod Brian Jimenez, Albert E Schmelzer, Michael C Jewett, Varnika Roy
Chinese Hamster Ovary (CHO) cells are routinely optimized to stably express monoclonal antibodies (mAbs) at high titers. At the early stages of lead isolation and optimization, hundreds of sequences for the target protein of interest are screened. Typically, cell-based transient expression technology platforms are used for expression screening, but these can be time- and resource-intensive. Here, we have developed a cell-free protein synthesis (CFPS) platform utilizing a commercially available CHO extract for the rapid in vitro synthesis of active, aglycosylated mAbs...
March 28, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28346939/a-b12-dependent-radical-sam-enzyme-involved-in-oxetanocin-a-biosynthesis
#2
Jennifer Bridwell-Rabb, Aoshu Zhong, He G Sun, Catherine L Drennan, Hung-Wen Liu
Oxetanocin A (OXT-A) is a potent antitumour, antiviral and antibacterial compound. Biosynthesis of OXT-A has been linked to a plasmid-borne Bacillus megaterium gene cluster that contains four genes: oxsA, oxsB, oxrA and oxrB. Here we show that both the oxsA and oxsB genes are required for the production of OXT-A. Biochemical analysis of the encoded proteins, a cobalamin (Cbl)-dependent S-adenosylmethionine (AdoMet) radical enzyme, OxsB, and an HD-domain phosphohydrolase, OxsA, reveals that OXT-A is derived from a 2'-deoxyadenosine phosphate in an OxsB-catalysed ring contraction reaction initiated by hydrogen atom abstraction from C2'...
March 27, 2017: Nature
https://www.readbyqxmd.com/read/28346803/carbapenemase-producing-aeromonas-veronii-disseminated-in-the-environment-of-an-equine-specialty-hospital
#3
Rachael J Adams, Dimitria A Mathys, Dixie F Mollenkopf, Andrea Whittle, Joshua B Daniels, Thomas E Wittum
The emergence of bacterial resistance to carbapenem antibiotics is an urgent public health threat. Carbapenem drugs are a last resort treatment option for life-threatening infections. The frequent use of broad-spectrum antibiotics to treat hospitalized patients provides significant selection pressure favoring the emergence and dissemination of resistant organisms, including carbapenem-resistant Enterobacteriaceae (CRE). CREs have been reported in animal populations, but only rarely in horses. Our objective was to determine the prevalence of CRE in the environment of a referral equine specialty hospital...
March 27, 2017: Vector Borne and Zoonotic Diseases
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
#4
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
https://www.readbyqxmd.com/read/28344787/allele-specific-transcriptional-activity-of-the-variable-number-of-tandem-repeats-of-the-inducible-nitric-oxide-synthase-gene-is-associated-with-idiopathic-achalasia
#5
Giovanni Sarnelli, Michela Grosso, Ilaria Palumbo, Marcella Pesce, Alessandra D'Alessandro, Giovanni Zaninotto, Vito Annese, Raffaella Petruzzelli, Paola Izzo, Rossana Sepulveres, Dario Bruzzese, Giuseppe Esposito, Rosario Cuomo
BACKGROUND: Polymorphisms of genes involved in the regulation of the immune response are risk factors for achalasia, but their contribution to disease pathogenesis is unknown. Nitric oxide is involved both in immune function and inhibitory neurotransmission. OBJECTIVE: The objective of this article is to assess the association and the functional relevance of the CCTTT-inducible nitric oxide synthase (NOS2) gene promoter polymorphism in achalasia. METHODS: Genomic DNA was isolated from 181 achalasia patients and 220 controls...
March 2017: United European Gastroenterology Journal
https://www.readbyqxmd.com/read/28344773/escherichia-coli-st131-a-multidrug-resistant-clone-primed-for-global-domination
#6
REVIEW
Johann D D Pitout, Rebekah DeVinney
A single extra-intestinal pathogenic Escherichia coli (ExPEC) clone, named sequence type (ST) 131, is responsible for millions of global antimicrobial-resistant (AMR) infections annually. Population genetics indicate that ST131 consists of different clades (i.e. A, B, and C); however, clade C is the most dominant globally. A ST131 subclade, named C1-M27, is emerging in Japan and has been responsible for the recent increase in AMR ExPEC in that country. The sequential acquisition of several virulence and AMR genes associated with mobile genetic elements during the 1960s to 1980s primed clade C (and its subclades C1 and C2) for success in the 1990s to 2000s...
2017: F1000Research
https://www.readbyqxmd.com/read/28344161/cross-resistance-to-human-cationic-antimicrobial-peptides-and-to-polymyxins-mediated-by-the-plasmid-encoded-mcr-1
#7
Jan Dobias, Laurent Poirel, Patrice Nordmann
OBJECTIVES: To evaluate whether acquired resistance to cationic antimicrobial peptides (CAMP) group molecules, being normal components of the human immune system, may select co-resistance to antibiotic peptides such as polymyxins, considering they share the same mechanism of action. We aimed to evaluate strains producing the recently identified plasmid-encoded polymyxin resistance determinant MCR-1, which is a phosphoethanolamine transferase that modifies the lipopolysaccharide structure of Gram-negative bacteria...
March 23, 2017: Clinical Microbiology and Infection
https://www.readbyqxmd.com/read/28344092/the-over-expression-of-long-non-coding-rna-anril-promotes-epithelial-mesenchymal-transition-by-activating-the-atm-e2f1-signaling-pathway-in-pancreatic-cancer-an-in-vivo-and-in-vitro-study
#8
Shi Chen, Jia-Qiang Zhang, Jiang-Zhi Chen, Hui-Xing Chen, Fu-Nan Qiu, Mao-Lin Yan, Yan-Ling Chen, Cheng-Hong Peng, Yi-Feng Tian, Yao-Dong Wang
This study aims to investigate the roles of lncRNA ANRIL in epithelial-mesenchymal transition (EMT) by regulating the ATM-E2F1 signaling pathway in pancreatic cancer (PC). PC rat models were established and ANRIL overexpression and interference plasmids were transfected. The expression of ANRIL, EMT markers (E-cadherin, N-cadherin and Vimentin) and ATM-E2F1 signaling pathway-related proteins (ATM, E2F1, INK4A, INK4B and ARF) were detected. Small molecule drugs were applied to activate and inhibit the ATM-E2F1 signaling pathway...
March 23, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28343162/mesothelin-promoter-variants-are-associated-with-increased-soluble-mesothelin-related-peptide-levels-in-asbestos-exposed-individuals
#9
Chiara De Santi, Perla Pucci, Alessandra Bonotti, Ombretta Melaiu, Monica Cipollini, Roberto Silvestri, Veronika Vymetalkova, Elisa Barone, Elisa Paolicchi, Alda Corrado, Irene Lepori, Irene Dell'Anno, Lucia Pellè, Pavel Vodicka, Luciano Mutti, Rudy Foddis, Alfonso Cristaudo, Federica Gemignani, Stefano Landi
BACKGROUND: Soluble mesothelin-related peptide (SMRP) is a promising diagnostic biomarker for malignant pleural mesothelioma (MPM), but various confounders hinder its usefulness in surveillance programmes. We previously showed that a single nucleotide polymorphism (SNP) within the 3'untranslated region (3'UTR) of the mesothelin (MSLN) gene could affect the levels of SMRP. OBJECTIVES: To focus on SNPs located within MSLN promoter as possible critical genetic variables in determining SMRP levels...
March 25, 2017: Occupational and Environmental Medicine
https://www.readbyqxmd.com/read/28342941/is-there-any-crosstalk-between-the-chemotaxis-and-virulence-induction-signaling-in-agrobacterium-tumefaciens
#10
REVIEW
Minliang Guo, Zhiwei Huang, Jing Yang
Agrobacterium tumefaciens, a soil-born phytopathogenic bacterium, is well known as a nature's engineer due to its ability to genetically transform the host by transferring a DNA fragment (called T-DNA) from its Ti plasmid to host-cell genome. To combat the harsh soil environment and seek the appropriate host, A. tumefaciens can sense and be attracted by a large number of chemical compounds released by wounded host. As a member of α-proteobacterium, A. tumefaciens has a chemotaxis system different from that found in Escherichia coli, since many chemoattractants for A...
March 23, 2017: Biotechnology Advances
https://www.readbyqxmd.com/read/28342852/effects-of-sub-culturing-on-genetic-and-physiological-parameters-in-different-beauveria-bassiana-isolates
#11
Naser Eivazian Kary, Zhila Alizadeh
Beauveria bassiana is a fungus which is widely used as a biological insecticide to control a number of economically important insect pests. Knowledge of the genetic diversity of the isolates, understanding the underlying nature of these evolutionary phenomena and finding appropriate and simple screening tools play an important role in developing effective biocontrol agents. Here, we monitored changes of electrophoretic karyotype of small molecules of extrachromosomal DNAs, presumably mitochondrial DNA or plasmids in several individual isolates of B...
March 22, 2017: Journal of Invertebrate Pathology
https://www.readbyqxmd.com/read/28341962/tumour-suppressor-ep300-a-modulator-of-paclitaxel-resistance-and-stemness-is-downregulated-in-metaplastic-breast-cancer
#12
Muhammad Asaduzzaman, Stephanie Constantinou, Haoxiang Min, John Gallon, Meng-Lay Lin, Poonam Singh, Selina Raguz, Simak Ali, Sami Shousha, R Charles Coombes, Eric W-F Lam, Yunhui Hu, Ernesto Yagüe
PURPOSE: We have previously described a novel pathway controlling drug resistance, epithelial-to-mesenchymal transition (EMT) and stemness in breast cancer cells. Upstream in the pathway, three miRs (miR-106b, miR-93 and miR-25) target EP300, a transcriptional activator of E-cadherin. Upregulation of these miRs leads to the downregulation of EP300 and E-cadherin with initiation of an EMT. However, miRs regulate the expression of many genes, and the contribution to EMT by miR targets other than EP300 cannot be ruled out...
March 24, 2017: Breast Cancer Research and Treatment
https://www.readbyqxmd.com/read/28341681/chromosome-encoded-hemolysin-phospholipase-and-collagenase-contribute-to-virulence-for-fish-in-plasmidless-isolates-of-photobacterium-damselae-subsp-damselae
#13
Ana Vences, Amable J Rivas, Manuel L Lemos, Matthias Husmann, Carlos R Osorio
Photobacterium damselae subsp. damselae is a pathogen of marine animals including fish of importance in aquaculture. The virulence plasmid pPHDD1, characteristic of highly hemolytic isolates, encodes the hemolysins damselysin (Dly) and phobalysin (PhlyP). Strains lacking pPHDD1 constitute the vast majority of the isolates from fish outbreaks, but genetic studies to identify virulence factors in plasmidless strains are scarce. Here we show that the chromosome-I encoded hemolysin PhlyC plays a role in virulence and cell toxicity in pPHDD1-negative isolates of this pathogen...
March 24, 2017: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/28341670/prime-boost-immunization-eliminates-metastatic-colorectal-cancer-by-producing-high-avidity-effector-cd8-t-cells
#14
Bo Xiang, Trevor R Baybutt, Lisa Berman-Booty, Michael S Magee, Scott A Waldman, Vitali Y Alexeev, Adam E Snook
Heterologous prime-boost immunization with plasmid DNA and viral vector vaccines is an emerging approach to elicit CD8(+) T cell-mediated immunity targeting pathogens and tumor Ags that is superior to either monotherapy. Yet, the mechanisms underlying the synergy of prime-boost strategies remain incompletely defined. In this study, we examine a DNA and adenovirus (Ad5) combination regimen targeting guanylyl cyclase C (GUCY2C), a receptor expressed by intestinal mucosa and universally expressed by metastatic colorectal cancer...
March 24, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28339845/expanded-spectrum-%C3%AE-lactamase-producing-escherichia-coli-isolated-from-chickens-with-colibacillosis-in-egypt
#15
D A El-Shazly, S A Nasef, F F Mahmoud, Daniel Jonas
Throughout the world, expanded spectrum β-lactamases (ESBL) are increasing among clinical isolates of Enterobacteriaceae, both in humans and animals. Unfortunately, there is a paucity of data on ESBL or Ampicillin class C β-lactamase (AmpC) in Egypt, although antimicrobial consumption is high in this developing country. This study aims to characterize the resistance mechanisms to expanded spectrum cephalosporins among resistant veterinary Escherichia coli isolates in Egypt. We investigated 50 clinical multi-resistant E...
February 23, 2017: Poultry Science
https://www.readbyqxmd.com/read/28339056/nobiletin-inhibits-invasion-via-inhibiting-akt-gsk3%C3%AE-%C3%AE-catenin-signaling-pathway-in-slug-expressing-glioma-cells
#16
Xiang Zhang, Kebin Zheng, Chunhui Li, Yonghui Zhao, Heyang Li, Xuguang Liu, Yinbo Long, Junchao Yao
Epithelial-mesenchymal transition (EMT) is a pivotal event in tumor progression during which cancer cells undergo dramatic changes acquiring highly invasive properties. In this study, we found that nobiletin, a polymethoxylated flavone, suppressed migration and invasion in both U87 and U251 glioma cells. Expression of epithelial markers (E-cadherin and occludin) was upregulated; mesenchymal markers (N-cadherin, fibronectin) and the transcriptional factor Slug were downregulated after nobiletin treatment. Transforming growth factor β (TGF-β) was applied to stimulate EMT and the results showed that nobiletin not only influenced basal level cell migration but also prevented TGF-β-triggered migration and EMT, with the AKT/GSK3β/β-catenin signaling pathway greatly involved...
March 23, 2017: Oncology Reports
https://www.readbyqxmd.com/read/28339048/agr2-ameliorates-tumor-necrosis-factor-%C3%AE-induced-epithelial-barrier-dysfunction-via-suppression-of-nf-%C3%AE%C2%BAb-p65-mediated-mlck-p-mlc-pathway-activation
#17
Xiaolin Ye, Mei Sun
Intestinal epithelial barrier dysfunction plays a critical role in the pathogenesis of inflammatory bowel disease (IBD). Anterior gradient protein 2 homologue (AGR2) assists in maintaining intestinal homeostasis in dextran sulphate sodium-induced mouse ileocolitis; however, it is unclear whether it modulates intestinal barrier function. Our study aimed to investigate the protective role of AGR2 in tumor necrosis factor (TNF)-α-induced intestinal epithelial barrier injury. Caco-2 cell monolayers were pre-transfected with an AGR2 plasmid and then exposed to TNF-α...
March 21, 2017: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/28338809/resveratrol-derivative-btm-0512-mitigates-obesity-by-promoting-beige-remodeling-of-subcutaneous-preadipocytes
#18
Qingqing Li, Kuansong Wang, Yipeng Ma, Chunxiang Qin, Changsheng Dong, Ping Jin, Yan Wu, Xiaoming Xiong, Niansheng Li, Changping Hu, Jun Peng, Zhichun Yang
Recent studies revealed that sirtuin 1 (SIRT1) is involved in the regulation of energy metabolism and its agonist resveratrol showed anti-obesity effect. This study aims to determine whether BTM-0512, a novel derivative of resveratrol, acts as an antagonist of obesity and to explore its possible mechanisms. High-fat diet (HFD)-induced obese mice were intragastrically administered with BTM-0512 (5, 10, and 20 mg/kg/day) or resveratrol (10 mg/kg/day). It was found that the body weight, Lee's index, ratio of visceral adipose tissue (VAT) to body weight, and blood glucose were significantly reduced in BTM-0512-treated mice when compared with those in mice treated with resveratrol...
February 24, 2017: Acta Biochimica et Biophysica Sinica
https://www.readbyqxmd.com/read/28338777/the-chlamydia-suis-genome-exhibits-high-levels-of-diversity-plasticity-and-mobile-antibiotic-resistance-comparative-genomics-of-a-recent-livestock-cohort-shows-influence-of-treatment-regimes
#19
Helena M B Seth-Smith, Sabrina Wanninger, Nathan Bachmann, Hanna Marti, Weihong Qi, Manuela Donati, Antonietta di Francesco, Adam Polkinghorne, Nicole Borel
Chlamydia suis is an endemic pig pathogen, belonging to a fascinating genus of obligate intracellular pathogens. Of particular interest, this is the only chlamydial species to have naturally acquired genes encoding for tetracycline resistance. To date, the distribution and mobility of the Tet-island is not well understood. Our study focused on whole genome sequencing of 29 C. suis isolates from a recent porcine cohort within Switzerland, combined with data from USA tetracycline-resistant isolates. Our findings show that the genome of C...
March 2, 2017: Genome Biology and Evolution
https://www.readbyqxmd.com/read/28338198/the-expression-of-sirt3-in-primary-hepatocellular-carcinoma-and-the-mechanism-of-its-tumor-suppressing-effects
#20
Y Liu, Y-L Liu, W Cheng, X-M Yin, B Jiang
OBJECTIVE: To observe the SIRT 3 expression in primary hepatocellular carcinoma (HCC), and then establish the eukaryotic expression vector of SIRT3 to observe the proliferation and apoptosis of pZsGreen-c1-SIRT3 HepG2 cells. Furthermore, we explored the mechanism of SIRT3 in inhibiting HCC. PATIENTS AND METHODS: Immunohistochemistry was used to detect the expression of SIRT3 in the tumor tissue and para-tumor tissue in 32 patients with HCC and the normal liver tissue in 10 patients...
March 2017: European Review for Medical and Pharmacological Sciences
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