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https://www.readbyqxmd.com/read/28637001/a-novel-method-to-generate-salmonella-typhi-ty21a-ghosts-exploiting-the-%C3%AE-phage-holin-endolysin-system
#1
Gayeon Won, Boram Kim, John Hwa Lee
Human typhoid fever caused by Salmonella Typhi still poses a severe global disease burden in developing countries despite the availability of commercial vaccines. In this study, we constructed a non-living S. Typhi Ty21a vaccine candidate by employing a lambda (λ) phage-derived holin-endolysin system to efficiently construct bacterial ghosts. The lysis plasmid pJHL464 harbors an R lysis cassette that is stringently regulated by dual promoters containing cI857/λPR and ParaBAD/araC components. The plasmid was introduced into an asd gene-deleted S...
June 6, 2017: Oncotarget
https://www.readbyqxmd.com/read/28636172/a-phos-tag-based-micropipette-tip-method-for-rapid-and-selective-enrichment-of-phosphopeptides
#2
Elena Tianfei Yuan, Yoko Ino, Maho Kawaguchi, Yayoi Kimura, Hisashi Hirano, Emiko Kinoshita-Kikuta, Eiji Kinoshita, Tohru Koike
Phosphorylated peptides are attractive targets in the study of the phosphoproteome. Here, we introduce a simple and convenient micropipette-tip method for the separation of phosphorylated and nonphosphorylated peptides by using a phosphate-binding zinc(II) complex of 1,3-bis(pyridin-2-ylmethylamino)propan-2-olate (Phos-tag). A 200-μL micropipette tip containing 10 μL of swollen agarose beads functionalized with Phos-tag moieties was prepared. All steps in the phosphate-affinity separation (binding, washing, and elution) were conducted by using aqueous buffers at neutral pH values...
June 21, 2017: Electrophoresis
https://www.readbyqxmd.com/read/28635254/sudoku-like-lab-on-paper-cyto-device-with-dual-enhancement-of-electrochemiluminescence-intermediates-strategy
#3
Hongmei Yang, Yan Zhang, Li Li, Lina Zhang, Feifei Lan, Jinghua Yu
This paper describes the design and construction of sudoku-like lab-on-paper platform, in which dual enhancement of reaction intermediates strategy was incorporated for multiplexed competitive electrochemiluminescence (ECL) cyto-assay. Benefit from the sudoku-like structure, integrated multifunction were obtained on such elaborately devised platform, including specific reagents storage, multiple samples immobilization, residual automatic washing, and signal collection. By utilizing semicarbazide (SE) and silver nanoparticles (AgNPs) as dual enhancers, more ECL intermediates could be obtained in the graphene quantum dots (GQDs) and peroxydisulfate system, resulting in the production of more excited state GQDs to emit light...
June 21, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28631587/dietary-supplementation-with-a-nucleotide-rich-yeast-extract-modulates-gut-immune-response-and-microflora-in-weaned-pigs-in-response-to-a-sanitary-challenge
#4
S M Waititu, F Yin, R Patterson, A Yitbarek, J C Rodriguez-Lecompte, C M Nyachoti
An experiment was carried out to evaluate the short-term effect of supplementing a nucleotide-rich yeast extract (NRYE) on growth performance, gut structure, immunity and microflora of piglets raised under sanitary and unsanitary conditions. A total of 84, 21-day old piglets were used in this study; 42 piglets were raised in a room designated as the clean room that was washed once per week, whereas the other 42 piglets were raised in a room designated as the unclean room in which 7 kg of manure from the sow herd was spread on each pen floor on day 1 and 7 and the room was not washed throughout the experiment...
June 20, 2017: Animal: An International Journal of Animal Bioscience
https://www.readbyqxmd.com/read/28630286/efficient-switching-of-mcherry-fluorescence-using-chemical-caging
#5
Bas M C Cloin, Elke De Zitter, Desiree Salas, Vincent Gielen, Gert E Folkers, Marina Mikhaylova, Maike Bergeler, Bartosz Krajnik, Jeremy Harvey, Casper C Hoogenraad, Luc Van Meervelt, Peter Dedecker, Lukas C Kapitein
Fluorophores with dynamic or controllable fluorescence emission have become essential tools for advanced imaging, such as superresolution imaging. These applications have driven the continuing development of photoactivatable or photoconvertible labels, including genetically encoded fluorescent proteins. These new probes work well but require the introduction of new labels that may interfere with the proper functioning of existing constructs and therefore require extensive functional characterization. In this work we show that the widely used red fluorescent protein mCherry can be brought to a purely chemically induced blue-fluorescent state by incubation with β-mercaptoethanol (βME)...
June 19, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28621606/nano-targeting-of-mucosal-addressin-cell-adhesion-molecule-1-identifies-bowel-inflammation-foci-in-murine-model
#6
Marta Truffi, Miriam Colombo, Jesus Peñaranda-Avila, Luca Sorrentino, Francesco Colombo, Matteo Monieri, Veronica Collico, Pietro Zerbi, Erika Longhi, Raffaele Allevi, Davide Prosperi, Fabio Corsi
AIM: We investigate MAdCAM-1 as a reliable target to detect active bowel inflammation for selective noninvasive nanodiagnostics. MATERIALS & METHODS: We coupled anti-MAdCAM-1 antibodies to manganese oxide nanoparticles, and analyzed nanoconjugate biodistribution and safety in murine model of inflammatory bowel disease by imaging and histology. RESULTS: Nanoparticles were stable and nontoxic. Upon administration in colitic mice, anti-MAdCAM-1 functionalized nanoparticles preferentially localized in the inflamed bowel, whereas untargeted nanoparticles were more rapidly washed out...
June 16, 2017: Nanomedicine
https://www.readbyqxmd.com/read/28620375/comparative-gene-expression-analysis-of-lymphocytes-treated-with-exosomes-derived-from-ovarian-cancer-and-ovarian-cysts
#7
Yujuan Li, Yang Yang, Aiwei Xiong, Xiaoqin Wu, Jingyan Xie, Suping Han, Shuli Zhao
Cancer cells employ many strategies to evade immune defense and to facilitate tumor growth and angiogenesis. As a novel mode of intercellular communication, cancer-derived exosomes contribute to the recruitment and mediation of lymphocytes within the tumor environment. However, the mechanisms and key molecules mediating the effect of exosomes on lymphocytes are unclear. We treated healthy peripheral blood lymphocytes with exosomes from ovarian cancer and ovarian cysts and screened for differentially expressed genes using the RT(2) Profiler Cancer Inflammation and Immunity Crosstalk PCR Array...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28618206/grazing-of-leaf-associated-cercomonads-protists-rhizaria-cercozoa-structures-bacterial-community-composition-and-function
#8
Sebastian Flues, David Bass, Michael Bonkowski
Preferential food selection in protists is well documented, but we still lack basic understanding on how protist predation modifies the taxonomic and functional composition of bacterial communities. We conducted feeding trials using leaf-associated cercomonad Cercozoa by incubating them on a standardized, diverse bacterial community washed from plant leaves. We used a shotgun metagenomics approach to investigate the taxonomic and functional changes of the bacterial community after five days protist predation upon bacteria...
June 15, 2017: Environmental Microbiology
https://www.readbyqxmd.com/read/28616254/in-vitro-ion-adsorption-and-cytocompatibility-of-dicalcium-phosphate-ceramics
#9
Martha Schamel, Jake E Barralet, Jürgen Groll, Uwe Gbureck
BACKGROUND: In vitro cell testing of degradable bioceramics such as brushite or monetite is often challenging due to the ion release into or adsorption from the culture medium. These ionic changes are then mostly responsible for cell proliferation and activity, which prohibits the investigation of effects originating from surface topography or further material modifications. METHODS: Here, we aimed to solve this problem by developing a pre-conditioning regime following the repeated immersion of brushite and monetite samples in various Ca(2+), Mg(2+) and PO4(3-) containing electrolytes, followed by studying ion adsorption / release as well as changes in phase composition and in vitro cytocompatibility with MG63 cells...
2017: Biomaterials Research
https://www.readbyqxmd.com/read/28613041/genotyping-horse-epithelial-cells-from-fecal-matter-by-isolation-of-polymerase-chain-reaction-products
#10
Pero Dimsoski
AIM: To show that application of the polymerase chain reaction (PCR) method modified for amplification of a low-copy number DNA samples, ie, the isolation of PCR products (IPCRp), would represent improvement in obtaining genotypes from a fecal DNA compared with previously used genotyping methods. METHODS: The DNA from the horse fecal matter was extracted by modified Qiagen DNA Stool Mini Kit protocol. Following the extraction, the DNA genotypes from fecal samples were obtained by the most powerful PCR amplification method, the IPCRp...
June 14, 2017: Croatian Medical Journal
https://www.readbyqxmd.com/read/28611099/cell-based-bioluminescent-assay-for-monitoring-the-interaction-between-pcsk9-and-the-ldl-receptor
#11
Sarah J Duellman, Thomas Machleidt, James J Cali, Jolanta Vidugiriene
Monitoring the expression of cell-surface receptors, their interaction with extracellular ligands, and their fate upon ligand binding is important for understanding receptor function and developing new therapies. We describe a cell-based method that utilizes bioluminescent protein complementation technology to interrogate binding of a cellular receptor with its extracellular protein ligand, specifically LDLR and PCSK9. Purified, full-length tagged PCSK9 is added to assay wells containing cells that stably express LDLR with an extracellular complementary tag...
June 13, 2017: Journal of Lipid Research
https://www.readbyqxmd.com/read/28606862/pathogenicity-profile-of-vibrio-parahaemolyticus-in-farmed-pacific-white-shrimp-penaeus-vannamei
#12
R Ananda Raja, R Sridhar, C Balachandran, A Palanisammi, S Ramesh, K Nagarajan
A pathobiological study was conducted using Vibrio parahaemolyticus (VP) strain isolated from vibriosis affected shrimp (Penaeus vannamei) farms in Kancheepuram and Thiruvallur districts of Tamil Nadu during August 2014 to February 2015. The isolate was identified based on the morphological, physiological, biochemical and molecular characters. LD50 value with intramuscular injection was determined as 2.6 × 10(4) cfu/shrimp and sequential pathology was studied giving 6.1 × 10(3) cfu/shrimp (LD25). Total plate count (TPC) and total Vibrio count (TVC) in water, pond sediment, haemolymph, muscle, HP and gut were found significantly (P < 0...
June 9, 2017: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/28605368/murine-short-axis-ventricular-heart-slices-for-electrophysiological-studies
#13
Gabriel Peinkofer, Juergen Hescheler, Marcel Halbach
Murine cardiomyocytes have been extensively used for in vitro studies of cardiac physiology and new therapeutic strategies. However, multicellular preparations of dissociated cardiomyocytes are not representative of the complex in vivo structure of cardiomyocytes, non-myocytes and extracellular matrix, which influences both mechanical and electrophysiological properties of the heart. Here we describe a technique to prepare viable ventricular slices of adult mouse hearts with a preserved in vivo like tissue structure, and demonstrate their suitability for electrophysiological recordings...
June 4, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28601555/comparison-of-skin-decontamination-efficacy-of-commercial-decontamination-products-following-exposure-to-vx-on-human-skin
#14
L Thors, M Koch, E Wigenstam, B Koch, L Hägglund, A Bucht
The decontamination efficacy of four commercially available skin decontamination products following exposure to the nerve agent VX was evaluated in vitro utilizing a diffusion cell and dermatomed human skin. The products included were Reactive Skin Decontamination Lotion (RSDL), the Swedish decontamination powder 104 (PS104), the absorbent Fuller's Earth and the aqueous solution alldecontMED. In addition, various decontamination procedures were assessed to further investigate important mechanisms involved in the specific products, e...
June 7, 2017: Chemico-biological Interactions
https://www.readbyqxmd.com/read/28598683/buffered-romanowsky-giemsa-method-for-formalin-fixed-paraffin-embedded-sections-taming-a-traditional-stain
#15
D Stefanović, G Samardžija, A Redžek, M Arnaut, Z Nikin, M Stefanović
Romanowsky-Giemsa (RG) stains were devised during the 19th century for identifying plasmodia parasites in blood smears. Later, RG stains became standard procedures for hematology and cytology. Numerous attempts have been made to apply RG staining to formalin-fixed paraffin-embedded tissue sections, with varied success. Most published work on this topic described RG staining methods in which sections were overstained, then subjected to acid differentiation; unfortunately, the differentiation step often caused inconsistent staining outcomes...
June 9, 2017: Biotechnic & Histochemistry: Official Publication of the Biological Stain Commission
https://www.readbyqxmd.com/read/28596492/detection-of-tumor-derived-dna-dispersed-in-the-airway-improves-the-diagnostic-accuracy-of-bronchoscopy-for-lung-cancer
#16
Taichiro Goto, Yosuke Hirotsu, Takahiro Nakagomi, Daichi Shikata, Yujiro Yokoyama, Kenji Amemiya, Toshiharu Tsutsui, Yumiko Kakizaki, Toshio Oyama, Hitoshi Mochizuki, Yoshihiro Miyashita, Masao Omata
The diagnostic accuracy of bronchoscopy for detecting lung cancer, especially peripheral lung cancer with lesions outside the endoscopically visible range, remains unsatisfactory. The aim of this study was to perform next-generation sequencing on bronchoscopic specimens to determine whether this improves the accuracy of bronchoscopy for diagnosing lung cancer and to identify factors influencing sensitivity. The bronchoscopic sensitivity for diagnosing lung cancer was initially evaluated in 191 patients who underwent lobectomy after bronchoscopy at our hospital...
May 24, 2017: Oncotarget
https://www.readbyqxmd.com/read/28595132/functionality-based-method-for-simultaneous-isolation-of-rodent-hepatic-sinusoidal-cells
#17
L Stradiot, S Verhulst, T Roosens, C I Øie, I M Moya, G Halder, I Mannaerts, L A van Grunsven
Chronic liver disease is the result of long term exposure to viruses or toxins such as alcohol, fat and drugs, and forms the basis for the development of liver fibrosis and primary liver cancer. In vitro and in vivo models are key to study the pathways involved in chronic liver disease and for the development of therapeutics. 3D co-culture systems are becoming the in vitro standard, which requires freshly isolated primary hepatic cells. We developed a novel isolation method to simultaneously isolate liver sinusoidal endothelial cells (LSECs), Kupffer cells (KCs) and hepatic stellate cells (HSCs)...
September 2017: Biomaterials
https://www.readbyqxmd.com/read/28595095/differential-expression-between-dsp-only-and-dsp-pp523-transcripts-in-rat-molar-teeth
#18
Ya-Qin Zhu, Ryan M Song, Helena H Ritchie
OBJECTIVE: To compare the expression patterns of two multiple transcripts derived from DSP-PP gene during tooth development. One is DSP-only transcript (i.e. does not encode PP) and the other is DSP-PP523 transcript, a main DSP-PP transcript. DESIGN: Unique antisense and sense riboprobes were generated from DSP-only and DSPPP523 cDNAs for in situ studies to examine DSP-only and DSP-PP523 transcript expression in developing molars. Paraffin-embedded sections (5-7μ m) from embryonic 20day, postnatal 2, 3 and 6days were deparaffined and hydrated...
May 31, 2017: Archives of Oral Biology
https://www.readbyqxmd.com/read/28592624/prolonged-pemetrexed-infusion-plus-gemcitabine-in-refractory-metastatic-colorectal-cancer-preclinical-rationale-and-phase-ii-study-results
#19
Alessandro Passardi, Francesca Fanini, Livia Turci, Flavia Foca, Paola Rosetti, Silvia Ruscelli, Andrea Casadei Gardini, Martina Valgiusti, Claudio Dazzi, Maurizio Marangolo
LESSONS LEARNED: Difficulties in translating in vitro results into clinical practice are inevitable.Further efforts to verify the efficacy of alternative schedules of pemetrexed in solid tumors are encouraged. BACKGROUND: We investigated the cytotoxic activity of pemetrexed in combination with several drugs (gemcitabine, carboplatin, vinorelbine, and mitomycin C) using different exposure schedules in three colon cancer cell lines. The best results were obtained with the following schedule: a prolonged pemetrexed exposure followed by a 48-hour wash-out and then gemcitabine...
June 7, 2017: Oncologist
https://www.readbyqxmd.com/read/28591737/characteristics-of-ovarian-cancer-detection-by-a-near-infrared-fluorescent-probe-activated-by-human-nad-p-h-quinone-oxidoreductase-isozyme-1-hnqo1
#20
Yuko Nakamura, Zhenhua Shen, Toshiko Harada, Tadanobu Nagaya, Kazuhide Sato, Shuhei Okuyama, Fusa Ogata, Peter L Choyke, Robin L McCarley, Hisataka Kobayashi
Near-infrared (NIR) fluorescent probes are ideal for in vivo imaging, because they offer deeper tissue penetration by the light and lower background autofluorescence than fluorophores that emit in the visible range. Q3STCy is a newly synthesized, NIR light-emitting probe that is activated by an enzyme commonly overexpressed in tumor cells, human nicotinamide adenine dinucleotide (phosphate): quinone oxidoreductase isozyme 1, known as hNQO1 or DT-diaphorase. The purpose of this study is to compare the sensitivity of detecting peritoneal ovarian cancer metastasis (POCM) with Q3STCy and gGlu-HMRG, a green fluorescent probe, upon their surface application...
May 20, 2017: Oncotarget
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