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https://www.readbyqxmd.com/read/28441472/repurposing-a-two-component-system-based-biosensor-for-the-killing-of-vibrio-cholerae
#1
Premkumar Jayaraman, Maciej Bartosz Holowko, Jing Wui Yeoh, Sierin Lim, Chueh Loo Poh
New strategies to control cholera are urgently needed. This study develops an in vitro proof-of-concept sense-and-eradicate system in a wild-type Escherichia coli strain to target the causative pathogen Vibrio cholerae using a synthetic biology approach. Our engineered E. coli specifically detects V. cholerae via its quorum-sensing molecule CAI-1 and responds by expressing the lysis protein (YebF-Art-085), thereby self-lysing to release the killing protein (Art-085) to eradicate V. cholerae. For this report, we individually characterized the YebF-Art-085 and Art-085 expression and their activities when coupled to our previously developed V...
April 25, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28439826/surface-acoustic-wave-lysis-and-ion-exchange-membrane-quantification-of-exosomal-microrna
#2
Katherine E Richards, David B Go, Reginald Hill
MicroRNA detection and quantification are commonly explored techniques for diagnostic and prognostic predictions. Typically, microRNAs are extracted and purified from a biological source, converted into complementary DNA (cDNA), and amplified using real time polymerase chain reaction (RT-PCR). The number of RT-PCR cycles required to reach the threshold of detection provides a relative quantification of the target microRNA when this data is normalized to the quantity of a control microRNA. This methodology has several drawbacks, including the need to artificially amplify the target microRNA for detection as well as quantification errors that can occur due to expression level differences of the control microRNAs for normalization in various sample sources...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28439632/a-rat-hysteropexy-model-for-evaluating-adhesion-formation-and-comparison-of-two-different-structured-meshes
#3
Ayse Filiz Gokmen-Karasu, Serdar Aydin, Fatma Cavide Sonmez, Ilknur Adanir, Gulsah Ilhan, Seda Ates
INTRODUCTION AND HYPOTHESIS: Peritonization of mesh during sacrohysteropexy is generally advocated to prevent adhesions to the viscera; however, randomized clinical trials are lacking, and peritonization may not be completely possible in a laparoscopic hysteropexy procedure. Our main objective was to describe a basic experimental rat sacrohysteropexy model. We hypothesized that even when peritoneal closure was omitted, using composite mesh would result in less adhesions to the viscera...
April 24, 2017: International Urogynecology Journal
https://www.readbyqxmd.com/read/28439259/increased-biological-activity-of-aneurinibacillus-migulanus-strains-correlates-with-the-production-of-new-gramicidin-secondary-metabolites
#4
Faizah N Alenezi, Imen Rekik, Ali Chenari Bouket, Lenka Luptakova, Hedda J Weitz, Mostafa E Rateb, Marcel Jaspars, Stephen Woodward, Lassaad Belbahri
The soil-borne gram-positive bacteria Aneurinibacillus migulanus strain Nagano shows considerable potential as a biocontrol agent against plant diseases. In contrast, A. migulanus NCTC 7096 proved less effective for inhibition of plant pathogens. Nagano strain exerts biocontrol activity against some gram-positive and gram-negative bacteria, fungi and oomycetes through the production of gramicidin S (GS). Apart from the antibiotic effects, GS increases the rate of evaporation from the plant surface, reducing periods of surface wetness and thereby indirectly inhibiting spore germination...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28438599/newly-identified-invertebrate-type-lysozyme-splys-i-in-mud-crab-scylla-paramamosain-exhibiting-muramidase-deficient-antimicrobial-activity
#5
Jian Zhou, Shu Zhao, Wen-Hong Fang, Jun-Fang Zhou, Jing-Xiao Zhang, Hongyu Ma, Jiang-Feng Lan, Xin-Cang Li
Lysozymes are widely distributed immune effectors exerting muramidase activity against the peptidoglycan of the bacterial cell wall to trigger cell lysis. However, some invertebrate-type (i-type) lysozymes deficient of muramidase activity still exhibit antimicrobial activity. To date, the mechanism underlying the antimicrobial effect of muramidase-deficient i-type lysozymes remains unclear. Accordingly, this study characterized a novel i-type lysozyme, Splys-i, in the mud crab Scylla paramamosain. Splys-i shared the highest identity with the Litopenaeus vannamei i-type lysozyme (Lvlys-i2, 54% identity) at the amino acid level...
April 21, 2017: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/28436069/the-eprep-system-a-new-electrophoretic-approach-for-dna-isolation-from-biological-samples
#6
Corinna Kueppers, Sabine Schlappa, Annette Bogdoll, Lothar Breitkopf, Nicole Seip, Thorsten Singer
We present a new free-flow electrophoretic separation system that extends the established concepts of nucleic acid migration in an electric field to a broadly applicable preparative scale. The system comprises a disposable flow tube in which the target nucleic acids are separated from impurities by a balanced combination of electrophoretic migration and counter-streaming electroosmotic flow under the influence of an applied external electric field. Despite the complex theoretical background the introduced electrophoretic technology offers simple hardware setup and handling protocols...
April 24, 2017: Electrophoresis
https://www.readbyqxmd.com/read/28434980/thaw-and-use-target-cells-pre-labeled-with-calcein-am-for-antibody-dependent-cell-mediated-cytotoxicity-assays
#7
Shan Chung, Van Nguyen, Yuwen Linda Lin, Lynn Kamen, An Song
In vitro antibody-dependent cell-mediated cytotoxicity (ADCC) assays are routinely performed to support the research and development of therapeutic antibodies. In ADCC assays, target cells bound by the antibodies are lysed by activated effector cells following interactions between the Fc region of the bound antibody and Fcγ receptors on effector cells. Target cell lysis is typically measured by quantification of released endogenous enzymes, e.g., lactate dehydrogenase, or measurement of released exogenous labels, e...
April 20, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28433637/inhibition-of-pro-inflammatory-enzymes-by-medicinal-plants-from-the-argentinean-highlands-puna
#8
Romina Torres-Carro, María Inés Isla, Samanta Thomas-Valdes, Felipe Jiménez-Aspee, Guillermo Schmeda-Hirschmann, María Rosa Alberto
ETHNOPHARMACOLOGICAL RELEVANCE: Human groups in the Argentinean Andes highlands (Puna) selected native plants as anti-inflammatory agents. The indications of use are mainly to relieve pain, in infusions, ethanolic extracts or plasters. AIM OF THE STUDY: The objective of the study was to assess the effect of hydroalcoholic extracts from native highland plants as anti-inflammatory according to the traditional indications of use. The chemical profile of the three most active species was analyzed by HPLC-ESI-MS to get in insight into the constituents and the effects observed, according to the ethnopharmacological information...
April 19, 2017: Journal of Ethnopharmacology
https://www.readbyqxmd.com/read/28432442/smokeless-tobacco-products-harbor-diverse-bacterial-microbiota-that-differ-across-products-and-brands
#9
Eoghan M Smyth, Prachi Kulkarni, Emma Claye, Stephen Stanfill, Robert Tyx, Cynthia Maddox, Emmanuel F Mongodin, Amy R Sapkota
Smokeless tobacco products contain numerous chemical compounds, including known human carcinogens. Other smokeless tobacco constituents, including bacteria, may also contribute to adverse health effects among smokeless tobacco users. However, there is a lack of data regarding the microbial constituents of smokeless tobacco. Our goal was to characterize the bacterial microbiota of different smokeless tobacco products and evaluate differences across product types and brands. DNA was extracted from 15 brands of smokeless tobacco products (including dry snuff, moist snuff, snus, and Swedish snus) and 6 handmade products (e...
April 22, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28431999/novel-method-to-rapidly-and-efficiently-lyse-escherichia-coli-for-the-isolation-of-recombinant-protein
#10
Himanshu Joshi, Vikas Jain
Rapid and high-throughput protein purification methods are required to explore structure and function of several uncharacterized proteins. Isolation of recombinant protein expressed in Escherichia coli strain BL21 (DE3) depends largely on the efficient and speedy bacterial cell lysis, which is considered as the bottleneck during protein purification. Cells are usually lysed by either sonication or high pressure homogenization, both of which are slow, require special equipment, lead to heat generation, and may result in loss of protein's biological activity...
April 18, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28429726/verteporfin-induced-formation-of-protein-cross-linked-oligomers-and-high-molecular-weight-complexes-is-mediated-by-light-and-leads-to-cell-toxicity
#11
Eleni K Konstantinou, Shoji Notomi, Cassandra Kosmidou, Katarzyna Brodowska, Ahmad Al-Moujahed, Fotini Nicolaou, Pavlina Tsoka, Evangelos Gragoudas, Joan W Miller, Lucy H Young, Demetrios G Vavvas
Verteporfin (VP) was first used in Photodynamic therapy, where a non-thermal laser light (689 nm) in the presence of oxygen activates the drug to produce highly reactive oxygen radicals, resulting in local cell and tissue damage. However, it has also been shown that Verteporfin can have non-photoactivated effects such as interference with the YAP-TEAD complex of the HIPPO pathway, resulting in growth inhibition of several neoplasias. More recently, it was proposed that, another non-light mediated effect of VP is the formation of cross-linked oligomers and high molecular weight protein complexes (HMWC) that are hypothesized to interfere with autophagy and cell growth...
April 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28429138/prothrombotic-state-in-asthma-is-related-to-increased-levels-of-inflammatory-cytokines-il-6-and-tnf%C3%AE-in-peripheral-blood
#12
Stanislawa Bazan-Socha, Lucyna Mastalerz, Agnieszka Cybulska, Lech Zareba, Romy Kremers, Michal Zabczyk, Grazyna Pulka, Teresa Iwaniec, Coenraad Hemker, Anetta Undas
Recently, we have reported that asthma is associated with enhanced plasma thrombin formation and impaired fibrinolysis. The mechanisms underlying the prothrombotic state in this disease are unknown. Our aim was to investigate whether prothrombotic alterations in asthmatics are associated with inflammation. We studied 164 adult, white, stable asthmatics and 72 controls matched for age, sex, body mass index (BMI), and smoking. Plasma tumor necrosis factor α (TNFα), interleukin (IL)-6, and serum periostin were evaluated using ELISAs, and their associations with thrombin generation, fibrinolytic capacity, expressed as clot lysis time (CLT), and platelet markers were later analyzed...
April 20, 2017: Inflammation
https://www.readbyqxmd.com/read/28429059/a-novel-method-to-recover-inclusion-body-protein-from-recombinant-e-coli-fed-batch-processes-based-on-phage-%C3%AE-x174-derived-lysis-protein-e
#13
Daniela Ehgartner, Patrick Sagmeister, Timo Langemann, Andrea Meitz, Werner Lubitz, Christoph Herwig
Production of recombinant proteins as inclusion bodies is an important strategy in the production of technical enzymes and biopharmaceutical products. So far, protein from inclusion bodies has been recovered from the cell factory through mechanical or chemical disruption methods, requiring additional cost-intensive unit operations. We describe a novel method that is using a bacteriophage-derived lysis protein to directly recover inclusion body protein from Escherichia coli from high cell density fermentation process: The recombinant inclusion body product is expressed by using a mixed feed fed-batch process which allows expression tuning via adjusting the specific uptake rate of the inducing substrate...
April 20, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28428904/salvage-of-necrotic-appearing-limb-after-in-utero-endoscopic-lysis-of-constriction-bands
#14
M Abdel-Sattar, A Chon, B Chen, W Burkhalter, R H Chmait
We report a case of amniotic band syndrome complicated by constriction bands and marked distal swelling of both lower extremities. Color Doppler interrogation of the right lower extremity revealed complete lack of blood flow below the level of the constriction. Upon fetoscopic survey, the right lower extremity beyond the constriction band appeared dusky red and discolored with desquamation, consistent with a necrotic appearance. The constriction bands were cut in utero using endoshears, thereby allowing restoration of blood flow on postoperative day 1...
April 2017: American Journal of Perinatology Reports
https://www.readbyqxmd.com/read/28428785/interferon-gamma-induces-changes-in-natural-killer-nk-cell-ligand-expression-and-alters-nk-cell-mediated-lysis-of-pediatric-cancer-cell-lines
#15
Arianexys Aquino-López, Vladimir V Senyukov, Zlatko Vlasic, Eugenie S Kleinerman, Dean A Lee
Natural killer (NK) cells have therapeutic potential for cancer due to their capacity for targeting tumor cells without prior sensitization. Our laboratory has developed an NK cell expansion protocol that generates large quantities of NK cells for therapeutic infusion that secret 20 times the amount of interferon gamma (IFNγ) than resting NK cells. IFNγ can upregulate major histocompatibility complex (MHC)-class I, an inhibitory ligand for NK cells, but can also upregulate intercellular adhesion molecule 1 (ICAM-1) which promotes NK:target cell interaction for an efficient lysis...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28427157/the-anti-tumor-efficacy-of-3c23k-a-glyco-engineered-humanized-anti-misrii-antibody-in-an-ovarian-cancer-model-is-mainly-mediated-by-engagement-of-immune-effector-cells
#16
Pauline Estupina, Alexandre Fontayne, Jean-Marc Barret, Nathalie Kersual, Olivier Dubreuil, Marion Le Blay, Alexandre Pichard, Marta Jarlier, Martine Pugnière, Maëva Chauvin, Thierry Chardès, Jean-Pierre Pouget, Emmanuel Deshayes, Alexis Rossignol, Toufik Abache, Christophe de Romeuf, Aurélie Terrier, Lucie Verhaeghe, Christine Gaucher, Jean-François Prost, André Pèlegrin, Isabelle Navarro-Teulon
Ovarian cancer is the leading cause of death in women with gynecological cancers and despite recent advances, new and more efficient therapies are crucially needed. Müllerian Inhibiting Substance type II Receptor (MISRII, also named AMHRII) is expressed in most ovarian cancer subtypes and is a novel potential target for ovarian cancer immunotherapy. We previously developed and tested 12G4, the first murine monoclonal antibody (MAb) against human MISRII. Here, we report the humanization, affinity maturation and glyco-engineering steps of 12G4 to generate the Fc-optimized 3C23K MAb, and the evaluation of its in vivo anti-tumor activity...
February 24, 2017: Oncotarget
https://www.readbyqxmd.com/read/28426838/a-second-wave-of-salmonella-t3ss1-activity-prolongs-the-lifespan-of-infected-epithelial-cells
#17
Ciaran E Finn, Audrey Chong, Kendal G Cooper, Tregei Starr, Olivia Steele-Mortimer
Type III secretion system 1 (T3SS1) is used by the enteropathogen Salmonella enterica serovar Typhimurium to establish infection in the gut. Effector proteins translocated by this system across the plasma membrane facilitate invasion of intestinal epithelial cells. One such effector, the inositol phosphatase SopB, contributes to invasion and mediates activation of the pro-survival kinase Akt. Following internalization, some bacteria escape from the Salmonella-containing vacuole into the cytosol and there is evidence suggesting that T3SS1 is expressed in this subpopulation...
April 20, 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/28426158/a-novel-technique-for-decellularization-of-allogenic-nerves-and-in-vivo-study-of-their-use-for-peripheral-nerve-reconstruction
#18
F Boriani, N Fazio, C Fotia, L Savarino, N Nicoli Aldini, L Martini, N Zini, M Bernardini, N Baldini
Autografts represent the gold standard for peripheral nerve reconstruction but their limited availability, the discrepancy of nerve caliber and long surgical times are drawbacks. Allografts have therefore become a valid alternative option. In particular, acellular nerve allografts (ANAs) rather than fresh allografts do not need immunosuppression and appear to be safe and effective based on recent studies. An innovative method was conceived to obtain ANAs, so as to speed up nerve decellularization, without compromising nerve architecture, and without breaking the asepsis chain...
April 20, 2017: Journal of Biomedical Materials Research. Part A
https://www.readbyqxmd.com/read/28424941/isolation-and-complete-genome-sequence-of-a-novel-pseudoalteromonas-phage-ph357-from-the-yangtze-river-estuary
#19
Zheng Gong, Min Wang, Qingwei Yang, Zhongshi Li, Jun Xia, Yu Gao, Yong Jiang, Xue Meng, Zhaoyang Liu, Ding Yang, Fangfei Zhang, Hongbing Shao, Duobing Wang
Phage PH357, a novel lytic Pseudoalteromonas lipolytica phage belonging to the Myoviridae family was isolated from the Yangtze River estuary. The microbiological characterization demonstrated that phage PH357 is stable from -20 to 60 °C and the optimal pH 7. The one-step growth curve showed a latent period of 20 min, a rise period of 20 min, and the average burst size was about 85 virions per cell. Complete genome of phage PH357 was determined. Genome of phage PH357 consisted of a linear, double-stranded 136,203 bp DNA molecule with 34...
April 20, 2017: Current Microbiology
https://www.readbyqxmd.com/read/28424044/emergent-carotid-thromboendarterectomy-for-acute-symptomatic-occlusion-of-the-extracranial-internal-carotid-artery
#20
Igor Gunka, Dagmar Krajickova, Michal Lesko, Stanislav Jiska, Jan Raupach, Miroslav Lojik, Radovan Maly
BACKGROUND: Strokes secondary to acute internal carotid artery (ICA) occlusion are associated with an extremely poor prognosis. The best treatment approach in this setting is still unknown. The aim of our study was to evaluate the efficacy, safety, and outcomes of emergent surgical revascularization of acute extracranial ICA occlusion in patients with minor to severe ischemic stroke. METHODS: A retrospective analysis was performed using prospectively collected data of consecutive patients who underwent carotid thromboendarterectomy for symptomatic acute ICA occlusion during the period from January 2013 to December 2015...
May 2017: Vascular and Endovascular Surgery
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