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https://www.readbyqxmd.com/read/28816710/establishment-of-primary-cell-culture-from-ascitic-fluid-and-solid-tumor-obtained-from-epithelial-ovarian-carcinoma-patients
#1
Rajarshi Kar, Diwesh Chawla, Bindiya Gupta, Mohit Mehndiratta, Neelam Wadhwa, Rachna Agarwal
OBJECTIVE: Ovarian cancer is the seventh leading cause of cancer death worldwide. This is mainly due to late diagnosis and high rate of relapse and resistance following chemotherapy. In the present study, we describe simple and cost-effective method to establish primary culture from ascitic fluid and solid tumor obtained from epithelial ovarian carcinoma patient, which may provide a better tool for in vitro testing of drug sensitivity and designing individualized treatment protocol. METHODS: Complete Dulbecco modified Eagle medium (DMEM) was prepared by supplementing DMEM with 10% fetal bovine serum and antibiotics (ciprofloxacin and amphotericin B)...
August 16, 2017: International Journal of Gynecological Cancer
https://www.readbyqxmd.com/read/28815245/-experimental-study-on-osteogenesis-of-acellular-porcine-pericardium-in-guided-bone-regeneration
#2
Jun-Zhao Gao, Xue-Jian Zhang, Jia-Li Hou, Yue Deng, Yun-Peng Li
PURPOSE: To observe the retention time and effect of acellular porcine pericardium (APP) in bone regeneration for rabbit femoral bone defects. METHODS: APP was prepared as follows: fresh porcine pericardium was chosen, high and low osmotic NaCl solution were used to soak the specimen alternately, trypsin+EDTA were used for digest, and then immersed with TritonX-100, cross-linked by using glutaraldehyde. 24 New Zealand rabbits were selected, and the area of bone defect was created 8 mm×8 mm×5 mm in size on bilateral distal femur...
April 2017: Shanghai Kou Qiang Yi Xue, Shanghai Journal of Stomatology
https://www.readbyqxmd.com/read/28815010/development-and-characterisation-of-a-low-concentration-sodium-dodecyl-sulphate-decellularised-porcine-dermis
#3
Jack A Helliwell, Daniel S Thomas, Vaia Papathanasiou, Shervanthi Homer-Vanniasinkam, Amisha Desai, Louise M Jennings, Paul Rooney, John N Kearney, Eileen Ingham
The aim of this study was to adapt a proprietary decellularisation process for human dermis for use with porcine skin. Porcine skin was subject to: sodium chloride (1 M) to detach the epidermis, trypsin paste to remove hair follicles, peracetic acid (0.1% v/v) disinfection, washed in hypotonic buffer and 0.1% (w/v) sodium dodecyl sulphate in the presence of proteinase inhibitors followed by nuclease treatment. Cellular porcine skin, decellularised porcine and human dermis were compared using histology, immunohistochemistry, GSL-1 lectin (alpha-gal epitope) staining, biochemical assays, uniaxial tensile and in vitro cytotoxicity tests...
January 2017: Journal of Tissue Engineering
https://www.readbyqxmd.com/read/28805888/development-of-small-intestinal-enzyme-activities-and-their-relationship-with-some-gut-regulatory-peptides-in-grazing-sheep
#4
C L Wang, X Lang, P J Wu, D P Casper, F D Li
Growth depends on an animal's capacity to digest and assimilate ingested nutrients, and insufficient supply and impairment will constrain lamb growth. Eight groups of Alpine Finewool lambs were harvested on 0, 3, 7, 14, 21, 28, 42, and 56 d to measure pH and enzymatic activities in the duodenum, proximal jejunum, middle jejunum, distal jejunum, and ileum mucosa or digesta. From the duodenum to the ileum the pH of intestinal mucosa and digesta increased, whereas pH changed very little with age. The trypsin, chymotrypsin, lipase, lactase, and α-amylase activities observed at birth decreased by d 3, followed by a nonuniform enzymatic response in the small intestine...
August 2017: Journal of Animal Science
https://www.readbyqxmd.com/read/28802115/production-of-high-purity-alveolar-like-cells-from-ipscs-through-depletion-of-uncommitted-cells-after-afe-induction
#5
Adam Mitchell, Charles T Drinnan, Todd Jensen, Christine Finck
Protocols to differentiate induced pluripotent stem cells (iPSCs) into specialized cells are continually evolving. iPSCs can be differentiated to alveolar cells with protocols that focus on development, specifically by inducing differentiation into definitive endoderm (DE), anterior foregut endoderm (AFE) and then lung bud progenitor intermediaries. However, current protocols result in a relatively low yield of the desired alveolar cells. The aim of this study was to evaluate whether depleting uncommitted cells after AFE induction would have a beneficial effect on alveolar cell yield...
August 3, 2017: Differentiation; Research in Biological Diversity
https://www.readbyqxmd.com/read/28801839/biotransformation-with-cellulase-hemicellulase-and-yarrowia-lipolytica-boosts-health-benefits-of-okara
#6
Weng Chan Vong, Xin Ying Lim, Shao-Quan Liu
Okara (soybean residue) is a highly perishable food processing by-product from soymilk and tofu manufacture. It contains a large proportion of insoluble dietary fibre (40-60% on a dry basis), as well as digestion-resistant proteins, trypsin inhibitors and phytic acid. These factors contribute lead to the under-utilisation of okara. To improve the overall nutritional quality of okara, sequential saccharification of okara by Celluclast® 1.5L (cellulase) or Viscozyme® L (cellulase and hemicellulase) and fermentation by the yeast Yarrowia lipolytica were performed...
August 11, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28800539/development-of-a-rapid-and-sensitive-multiple-reaction-monitoring-proteomic-approach-for-quantification-of-transporters-in-human-liver-tissue
#7
Li Wang, Kasi J Rubadue, Jeffrey Alberts, David W Bedwell, Kenneth J Ruterbories
With increasing knowledge on the role of hepatic transporters in drug disposition, numerous efforts have been described to quantify the expression of human hepatic transporters. However, reported quantitative proteomic approaches often require long analysis times. Additionally, greater assay sensitivity is still necessary for less abundant transporters or limited quantity of samples (e.g. hepatocytes and liver tissue). In the present study, an LC-MS/MS method for rapid and simultaneous quantification of 12 hepatic transporters (BCRP, BSEP, MATE1, MRP2, MRP3, MRP4, NTCP, OATP1B1, 1B3, 2B1, OCT1, and P-gp) was developed...
August 1, 2017: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/28798909/analysis-of-urine-composition-in-type-ii-diabetic-mice-after-intervention-therapy-using-holothurian-polypeptides
#8
Yanyan Li, Jiajie Xu, Xiurong Su
Hydrolysates and peptide fractions (PF) obtained from sea cucumber with commercial enzyme were studied on the hyperglycemic and renal protective effects on db/db rats using urine metabolomics. Compared with the control group the polypeptides from the two species could significantly reduce the urine glucose and urea. We also tried to address the compositions of highly expressed urinary proteins using a proteomics approach. They were serum albumins, AMBP proteins, negative trypsin, elastase, and urinary protein, GAPDH, a receptor of urokinase-type plasminogen activator (uPAR), and Ig kappa chain C region...
2017: Frontiers in Chemistry
https://www.readbyqxmd.com/read/28798698/%C3%AE-catenin-knockdown-affects-mitochondrial-biogenesis-and-lipid-metabolism-in-breast-cancer-cells
#9
Daniele Vergara, Eleonora Stanca, Flora Guerra, Paola Priore, Antonio Gaballo, Julien Franck, Pasquale Simeone, Marco Trerotola, Stefania De Domenico, Isabelle Fournier, Cecilia Bucci, Michel Salzet, Anna M Giudetti, Michele Maffia
β-catenin plays an important role as regulatory hub in several cellular processes including cell adhesion, metabolism, and epithelial mesenchymal transition. This is mainly achieved by its dual role as structural component of cadherin-based adherens junctions, and as a key nuclear effector of the Wnt pathway. For this dual role, different classes of proteins are differentially regulated via β-catenin dependent mechanisms. Here, we applied a liquid chromatography-mass spectrometry (LC-MS/MS) approach to identify proteins modulated after β-catenin knockdown in the breast cancer cell line MCF-7...
2017: Frontiers in Physiology
https://www.readbyqxmd.com/read/28783436/a-multi-channel-open-tubular-enzyme-reactor-on-line-coupled-with-mass-spectrometry-for-detecting-ricin
#10
Ole Kristian Brandtzæg, Bent Tore Røen, Siri Enger, Elsa Lundanes, Steven Ray Wilson
For counter-terrorism purposes, a selective nano liquid chromatography-mass spectrometry (nanoLC-MS) platform was developed for detecting the highly lethal protein ricin from castor bean extract. Manual sample preparation steps were omitted by implementing a trypsin/Lys C enzyme-immobilized multi-channel reactor (MCR) consisting of 126 channels (8 µm inner diameter in all channels) that performed on-line digestion of proteins (5 minute reaction time, instead of 4-16 hours in previous in-solution methods). Reduction and alkylation steps were not required...
August 7, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28783371/molecular-characterization-and-biological-activity-of-interferon-%C3%AE-in-indian-peafowl-pavo-cristatus
#11
Hongjing Zhao, Yu Wang, Juanjuan Liu, Yizhi Shao, Jinglun Li, Hongliang Chai, Mingwei Xing
Interferon-α (IFN-α) genes have been cloned from a variety of animals, but information about Indian peafowl Pavo cristatus IFN-α (PcIFN-α) has not been reported. In this study, a PcIFN-α gene was amplified, which encoded a protein of 193 amino acids with a 26-amino acid signal peptide sharing 72.16-95.70% identity with other avians in Aves. After expression in prokaryote, purification, and renaturation, PcIFN-α was analyzed for its physicochemical property and antiviral activity. Cytopathic inhibition test showed that the recombinant PcIFN-α could inhibit the replication of vesicular stomatitis virus (VSV), Newcastle disease virus (NDV), and avian influenza virus (AIV) on chicken embryo fibroblasts...
August 7, 2017: DNA and Cell Biology
https://www.readbyqxmd.com/read/28783006/development-of-a-confirmatory-method-for-detecting-recombinant-bovine-somatotropin-in-plasma-by-immunomagnetic-precipitation-followed-by-ultra-high-performance-liquid-chromatography-coupled-to-tandem-mass-spectrometry
#12
Christelle Robert, Anne-Catherine Huet, Célia Suárez-Pantaleón, Amaury Brasseur, Philippe Delahaut, Nathalie Gillard
Recombinant bovine somatotropin (rbST), a synthetic growth hormone, is used to stimulate growth and enhance milk production in dairy cows. Both its use and the sale of dairy products from treated animals are prohibited in the European Union, Japan, Australia, New Zealand, and Canada, but authorised in several countries (e.g. the USA, Brazil). Screening methods involve detecting anti-rbST antibodies (biomarkers) in treated cows. Confirmatory methods are required to prove rbST abuse. The major challenges in determining rbST are its potentially low levels, its high similarity to native bST, and matrix interferences...
August 7, 2017: Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment
https://www.readbyqxmd.com/read/28781773/the-relationship-between-dietary-methionine-and-growth-digestion-absorption-and-antioxidant-status-in-intestinal-and-hepatopancreatic-tissues-of-sub-adult-grass-carp-ctenopharyngodon-idella
#13
Pei Wu, Ling Tang, Weidan Jiang, Kai Hu, Yang Liu, Jun Jiang, Shengyao Kuang, Ling Tang, Wuneng Tang, Yongan Zhang, Xiaoqiu Zhou, Lin Feng
BACKGROUND: Methionine is an essential amino acid for fish. The present study was conducted to investigate the effects of dietary methionine on growth performance, digestive and absorptive ability, as well as antioxidant capacity in the intestine and hepatopancreas of sub-adult grass carp (Ctenopharyngodon idella). RESULTS: Dietary methionine deficiency significantly decreased percentage weight gain (PWG), feed intake, feed efficiency and protein efficiency ratio, as well as activities of hepatopancreatic glutamate-oxaloacetate transaminase and muscle glutamate-pyruvate transaminase in sub-adult grass carp (P < 0...
2017: Journal of Animal Science and Biotechnology
https://www.readbyqxmd.com/read/28780598/optimization-of-monomethoxy-polyethyleneglycol-modified-oxalate-decarboxylase-by-response-surface-methodology
#14
Han Long, XingHua Cai, Hui Yang, JunBin He, Jia Wu, RiHui Lin
In order to improve the stability of oxalate decarboxylase (Oxdc), response surface methodology (RSM), based on a four-factor three-level Box-Behnken central composite design was used to optimize the reaction conditions of oxalate decarboxylase (Oxdc) modified with monomethoxy polyethyleneglycol (mPEG5000). Four independent variables such as the ratio of mPEG-aldehyde to Oxdc, reaction time, temperature, and reaction pH were investigated in this work. The structure of modified Oxdc was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier transform infrared (FTIR) spectroscopy, the stability of the modified Oxdc was also investigated...
August 5, 2017: Journal of Biological Physics
https://www.readbyqxmd.com/read/28780458/influence-of-an-external-electric-field-on-removal-of-protein-fouling-on-a-stainless-steel-surface-by-proteolytic-enzymes
#15
Ei Ei Htwe, Yuhi Nakama, Hiroyuki Imanaka, Naoyuki Ishida, Koreyoshi Imamura
Enzymatic cleaning is a potentially useful method for removing proteinaceous fouling from solid surfaces under mild conditions. Herein, the influence of an external electric field on the enzymatic cleaning of a metal surface fouled with a protein was investigated. The model fouling protein (BSA; lysozyme) was prepared on a stainless steel (St) surface, and the resulting surface subjected to enzymatic cleaning with an electric potential being applied to the St plate. Trypsin, α-chymotrypsin, and thermolysin were used as model proteases...
July 27, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28777955/supplementation-of-lactobacilli-improves-growth-regulates-microbiota-composition-and-suppresses-skeletal-anomalies-in-juvenile-pike-perch-sander-lucioperca-reared-in-recirculating-aquaculture-system-ras-a-pilot-study
#16
Uros Ljubobratovic, Dejana Kosanovic, Goran Vukotic, Zsuzsanna Molnar, Nemanja Stanisavljevic, Tijana Ristovic, Geza Peter, Jovanka Lukic, Galina Jeney
This research aimed to test the effects of lactobacilli, applied to cultured pike-perch, either through hydrolyzed OTOHIME fish diet, or through Artemia nauplii, on fish growth, microbiota balance and skeletal development. On the 12th Day Post Hatching (DPH) fish were divided into following treatment groups: two groups received the combination of OTOHIME and nauplii enriched either with Lactobacillus paracasei BGHN14+Lactobacillus rhamnosus BGT10 or with Lactobacillus reuteri BGGO6-55+Lactobacillus salivarius BGHO1, and one group received OTOHIME hydrolyzed by BGHN14+BGT10 and non-enriched nauplii...
July 24, 2017: Research in Veterinary Science
https://www.readbyqxmd.com/read/28777090/high-resolution-structure-of-a-kazal-type-serine-protease-inhibitor-from-the-dengue-vector-aedes-aegypti
#17
Ricardo J S Torquato, Stephen Lu, Nadia Helena Martins, Aparecida S Tanaka, Pedro José Barbosa Pereira
Blood-feeding exoparasites are rich sources of protease inhibitors, and the mosquito Aedes aegypti, which is a vector of Dengue virus, Yellow fever virus, Chikungunya virus and Zika virus, is no exception. AaTI is a single-domain, noncanonical Kazal-type serine proteinase inhibitor from A. aegypti that recognizes both digestive trypsin-like serine proteinases and the central protease in blood clotting, thrombin, albeit with an affinity that is three orders of magnitude lower. Here, the 1.4 Å resolution crystal structure of AaTI is reported from extremely tightly packed crystals (∼22% solvent content), revealing the structural determinants for the observed inhibitory profile of this molecule...
August 1, 2017: Acta Crystallographica. Section F, Structural Biology Communications
https://www.readbyqxmd.com/read/28776699/poly-n-acryloxysuccinimide-co-ethylene-glycol-dimethacrylate-precursor-monolith-and-its-post-polymerization-modification-with-alkyl-ligands-trypsin-and-lectins-for-reversed-phase-chromatography-miniaturized-enzyme-reactors-and-lectin-affinity-chromatography
#18
Murthy Jonnada, Ziad El Rassi
This investigation was aimed at introducing a monolithic precursor that can be conveniently grafted with the desired chromatographic ligand via the process of post polymerization modification (PPM). The precursor was obtained by the in-situ polymerization of N-acryloxysuccinimide (NAS) and ethylene glycol dimethacrylate (EDMA) in a narrow bore stainless steel column of 1 mm i.d. yielding a poly(NAS-co-EDMA) monolithic column designated as the NASM column. In a first PPM, the NASM column was bonded with octadecyl (OD) ligands yielding a nonpolar NASM-OD column that proved useful for RPC of proteins in gradient elution at increasing %ACN in the mobile phase...
August 4, 2017: Electrophoresis
https://www.readbyqxmd.com/read/28771242/pancreatic-and-intestinal-function-post-roux-en-y-gastric-bypass-surgery-for-obesity
#19
Stephen J D O'Keefe, Tina Rakitt, Junhai Ou, Ihab I El Hajj, Elizabeth Blaney, Kishore Vipperla, Jens-Jules Holst, Jens Rehlfeld
OBJECTIVES: Despite the fact that the most effective treatment for morbid obesity today is gastric bypass surgery, some patients develop life-threatening nutritional complications associated with their weight loss. METHODS: Here we examine the influence of the altered anatomy and digestive physiology on pancreatic secretion and fat absorption. Thirteen post Roux-en-Y gastric bypass (RYGB) patients who had lost >100 lbs in the first year following surgery and who gave variable histories of gastrointestinal (GI) dysfunction, were selected for study...
August 3, 2017: Clinical and Translational Gastroenterology
https://www.readbyqxmd.com/read/28769120/interferon-gamma-released-from-omental-adipose-tissue-of-insulin-resistant-humans-alters-adipocyte-phenotype-and-impairs-response-to-insulin-and-adiponectin-release
#20
J M Wentworth, J-G Zhang, E Bandala-Sanchez, G Naselli, C Liu, M Ritchie, G K Smyth, P E O'Brien, L C Harrison
BACKGROUND: Inflammatory factors derived from adipose tissue have been implicated in mediating insulin resistance in obesity. We sought to identify these using explanted human adipose tissue exposed to innate and adaptive immune stimuli. METHOD: Subcutaneous and omental adipose tissue from obese, insulin-resistant donors was cultured in the presence of macrophage and T-cell stimuli and the conditioned medium tested for its ability to inhibit insulin-stimulated glucose uptake into human Simpson-Golabi-Behmel Syndrome (SGBS) adipocytes...
August 3, 2017: International Journal of Obesity: Journal of the International Association for the Study of Obesity
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