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Bin Deng, Shaolong Zhu, Andrew M Macklin, Jianrong Xu, Cristina Lento, Adnan Sljoka, Derek J Wilson
Localization of the interface between the candidate antibody and its antigen target, commonly known as epitope mapping, is a critical component of the development of therapeutic monoclonal antibodies. With the recent availability of commercial automated systems, hydrogen / deuterium eXchange (HDX) is rapidly becoming the tool for mapping epitopes preferred by researchers in both industry and academia. However, this approach has a significant drawback in that it can be confounded by 'allosteric' structural and dynamic changes that result from the interaction, but occur far from the point(s) of contact...
September 21, 2017: MAbs
Hui-Fang Ji, Lu Liu, Kai Li, Yue-Yuan Zhang, Ling-Zhi Wang
To analyze the protein composition of Brucea javanica seeds and evaluate the cytotoxicity of its gulbulin hydrolysates. Four protein fractions of albumin, gulbulin, prolamin and glutelin were sequentially extracted and then quantified by Kjeldahl method. Different kinds of proteases were applied to hydrolyze B.javanica gulbulin, and MTT assay was used to evaluate the cytotoxicity of low molecular weight hydrolysates (≤3 kDa) on human breast cancer MCF-7 cell. The results showed that: the total protein content of B...
November 2016: Zhongguo Zhong Yao za Zhi, Zhongguo Zhongyao Zazhi, China Journal of Chinese Materia Medica
Antonio Ortega-Martinez, Genna Touchette, Hong Zhu, Irene E Kochevar, Walfre Franco
Keratoconus is an eye disease in which the cornea progressively deforms due to loss of cornea mechanical rigidity, and thus causes deterioration of visual acuity. Techniques to characterize the mechanical characteristics of the cornea are important to better monitor changes and response to treatments. To investigate the feasibility of using the endogenous fluorescence of cornea for monitoring alterations of its mechanical rigidity, linear tensiometry was used to quantitate stiffness and Young's modulus (YM) after treatments that increase cornea stiffness (collagen photocross-linking) or decrease stiffness (enzymatic digestion)...
September 2017: Journal of Biomedical Optics
Chiara Zanoni, Gilda Aiello, Anna Arnoldi, Carmen Lammi
This study had the objective of preparing a hempseed protein hydrolysate and investigating its hypocholesterolemic properties. The hydrolysate was prepared treating a total protein extract with pepsin. Nano HPLC-ESI-MS/MS analysis permitted identifying in total 90 peptides belonging to 33 proteins. In the range 0.1-1.0 mg/mL, it inhibited the catalytic activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCoAR) in a dose-dependent manner. HepG2 cells were treated with 0.25, 0.5, and 1.0 mg/mL of the hydrolysate...
September 20, 2017: Journal of Agricultural and Food Chemistry
Kosuke Otake, Norio Sato, Ayako Kitaguchi, Takayuki Irahara, Satoru Murata, Keiichiro Shiraga, Yuichi Ogawa, Takahiro K Fujiwara, Kaoru Koike, Hiroyuki Yokota
Clostridium difficile infections (CDI) have recently increased worldwide. Some CDI progress to fulminant and recurrent CDI and are associated with high mortality and morbidity. CD produces toxins A and B, which cause intestinal mucosal damage, although toxin B exhibits greater cytotoxicity. Pepsin-treated lactoferrin (PLF) is the decomposed product of lactoferrin (LF), a multifunctional glycoprotein with anti-inflammatory properties. Here, we investigate the effects of LF and PLF in toxin B-stimulated rat intestinal epithelial (IEC-6) cells...
September 19, 2017: Shock
Douglas G Hayes, Ran Ye, Rachel N Dunlap, Matthew J Cuneo, Sai Venkatesh Pingali, Hugh M O'Neill, Volker S Urban
Bicontinuous microemulsions (BμEs), consisting of water and oil nanodomains separated by surfactant monolayers of near-zero curvature, are potentially valuable systems for purification and delivery of biomolecules, for hosting multiphasic biochemical reactions, and as templating media for preparing nanomaterials. We formed Winsor-III systems by mixing aqueous protein and sodium dodecyl sulfate (SDS) solutions with dodecane and 1-pentanol (cosurfactant) to efficiently extract proteins into the middle (BμE) phase...
September 7, 2017: Colloids and Surfaces. B, Biointerfaces
Yanjun Liu, Shiaw-Lin Wu, Kerry Routenberg Love, William S Hancock
Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. Understanding site specific glycosylation of hemagglutinin will increase our knowledge about virus evolution and can improve the design and quality of vaccines. In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion were used to determine the glycosites and glycan occupancy...
September 18, 2017: Analytical Chemistry
Yuan Jin, Xiaoyun He, Kwame Andoh-Kumi, Rachel Z Fraser, Mei Lu, Richard E Goodman
SCOPE: The Soybean (Glycine max) leghemoglobin c2 (LegHb) gene was introduced into Pichia pastoris yeast for sustainable production of a heme-carrying protein, for organoleptic use in plant-based meat. The potential allergenicity and toxicity of LegHb and 17 Pichia host-proteins each representing ≥1% of total protein in production batches were evaluated by literature review, bioinformatics sequence comparisons to known allergens or toxins, and in vitro pepsin digestion. METHODS AND RESULTS: Literature searches found no evidence of allergenicity or toxicity for these proteins...
September 18, 2017: Molecular Nutrition & Food Research
Siyao Liu, Jeannie Horak, Markus Höldrich, Michael Lämmerhofer
The ability to accurately quantify the protein coverage on nanoparticles is critical for assessing the quality of the surface chemistry and the success of the functionalization process of protein-nanoparticle conjugates. Surface coverage determination is therefore an integral part in the quality control of protein-modified nanoparticles in industrial nanotechnology. In this work, a novel and conventional method was established for direct quantification of the protein surface coverage on metallic nanoparticles...
October 9, 2017: Analytica Chimica Acta
Wei Liao, Xia Guanghua, Yongcheng Li, Xuan Ri Shen, Chuan Li
The present study isolated and characterized the barramundi (Lates calcarifer) skin collagen (BC) and tilapia (Oreochromis niloticus) skin collagen (TC). The yields of BC and TC by enzymatic extraction were 47.3±3.7% and 52.6±6.1% respectively, dry weight. The SDS-PAGE profile indicated both collagens were mainly type I with two different α chains. FTIR spectra and X-ray diffraction confirmed that the triple helical structure of both collagens was not affected by pepsin digestion. The denaturation (Td) and melting temperature (Tm) were 36...
September 11, 2017: International Journal of Biological Macromolecules
Masoud Homayouni-Tabrizi, Ahmad Asoodeh, Mozhgan Soltani
Peptides from natural sources such as milk are shown to have a wide spectrum of biological activities. In this study, three peptides with antioxidant capacity were identified from camel milk protein hydrolysate. Pepsin and pancreatin were used for hydrolysis of milk proteins. Ultrafiltration and reverse-phase high-performance liquid chromatography were used for the concentration and purification of the hydrolysate, respectively. Sequences of the three peptides, which were determined by matrix-assisted laser desorption/ionization time-of-flight spectrophotometry, were LEEQQQTEDEQQDQL [molecular weight (MW): 1860...
July 2017: Journal of Food and Drug Analysis
Veronique Demers-Mathieu, Søren Drud Nielsen, Mark A Underwood, Robyn Borghese, David C Dallas
OBJECTIVES: Our previous studies suggested that human milk proteases begin to hydrolyze proteins in the mammary gland and continue within the term infant stomach. No research has measured milk protease and pepsin activity in the gastric aspirates of preterm infants after human milk feeding. This study investigated how the concentrations of human milk proteases and protease inhibitors changed in the premature infant stomach. METHODS: Human milk and infant gastric samples were collected from 18 preterm-delivering mother-infant pairs (24-32 wk gestational age)...
September 12, 2017: Journal of Pediatric Gastroenterology and Nutrition
Morteza Parvan, Sayed-Ebrahim Sajjadi, Mohsen Minaiyan
BACKGROUND: In various studies, Cydonia oblonga Miller (quince) has been reported to have many properties such as antioxidant and anti-ulcerative effects. This study has aimed to investigate the protective effects of quince aqueous extract (QAE) and quince hydroalcoholic extract (QHE) on gastric ulcer caused by indomethacin and the relevant macroscopic, histopathology, and biochemical factors in rats. METHODS: Ten groups of male Wistar rats, six in each, were used in this study...
2017: International Journal of Preventive Medicine
Beatriz Miralles, Roberto Del Barrio, Carolina Cueva, Isidra Recio, Lourdes Amigo
BACKGROUND: A dynamic gastrointestinal simulator, simgi(®) , has been applied to assess the gastric digestion of a whey protein concentrate. Samples collected from the outlet of the stomach have been compared to those resulting from the static digestion protocol INFOGEST developed on the basis of physiologically inferred conditions. RESULTS: Progress of digestion was followed by SDS-PAGE and LC-MS/MS. By SDS-PAGE, serum albumin and α-lactalbumin were no longer detectable at 30 and 60 min, respectively...
September 12, 2017: Journal of the Science of Food and Agriculture
Nabil Souissi, Ola Abdelhedi, Aïcha Mbarek, Wassim Kammoun, Hela Kechaou, Moncef Nasri
Gelatins from grey triggerfish skin were extracted with different methods The supplementation by pepsin (PG) improved the yield of extraction when compared with untreated gelatin (UG) and acidic gelatin (AG). The outputs of gelatins AG, UG and PG, obtained respectively, with acitic acid, glycine buffer and glycine buffer added with 5U of pepsin/g of the skin beforehand treated by alkali, were 6.9%, 7.9% and 9.7%, respectively. The enzymatic treatment of the alkali-pretreated skin of grey triggerfish altered the electrophoresis profile, biophysical, gellification, rheological and thermal properties of the prepared gelatins extracted under acidic condition...
September 8, 2017: International Journal of Biological Macromolecules
Alicia J Dafferner, Lawrence M Schopfer, Gaoping Xiao, John R Cashman, Udaya Yerramalla, Rudolph C Johnson, Thomas A Blake, Oksana Lockridge
Nerve agents and organophosphorus pesticides make a covalent bond with the active site serine of acetylcholinesterase (AChE), resulting in inhibition of AChE activity and toxic symptoms. AChE in red blood cells (RBC) serves as a surrogate for AChE in the nervous system. Mass spectrometry analysis of adducts on RBC AChE could provide evidence of exposure. Our goal was to develop a method of immunopurifying human RBC AChE in quantities adequate for detecting exposure by mass spectrometry. For this purpose we immobilized 3 commercially available anti-human acetylcholinesterase monoclonals (AE-1, AE-2, and HR2) plus 3 new monoclonals...
September 11, 2017: Chemical Research in Toxicology
Vince Beachley, Garret Ma, Chris Papadimitriou, Matt Gibson, Michael Corvelli, Jennifer Elisseeff
Tissue extracellular matrix (ECM) is a complex material made up of fibrous proteins and ground substance (glycosaminoglycans, GAGs) that are secreted by cells. ECM contains important biological cues that modulate cell behaviors, and it also serves as a structural scaffold to which cells can adhere. For clinical applications, where immune rejection is a constraint, ECM can be processed using decellularization methods intended to remove cells and donor antigens from tissue or organs, while preserving native biological cues essential for cell growth and differentiation...
September 6, 2017: Journal of Biomedical Materials Research. Part A
Yang Li, Jing Li, Tianhong Liu, Ying Wang, Zhongzheng Zhou, Feng Cheng, Chao Feng, Xiaojie Cheng, Hongjun Liu, Xiguang Chen
In this work, a thermal and wide pH range (from 1 to 13) stable biological agent was extracted from Perinereis aibuhitensis (P. aibuhitensis), and the antithrombotic activity of this P. aibuhitensis extract (PaE) was investigated. In vitro antithrombotic activity test indicated that PaE could significantly extend the clotting time of recalcified rabbit blood: the clotting time of the 10 mg mL(-1), 20 mg mL(-1) and 40 mg mL(-1) PaE groups increased by 24.24%, 68.64% and 130.76%, respectively. Further platelet poor plasma (PPP) coagulation study proved that PaE could obviously inhibit the intrinsic coagulation pathway and slightly inhibit the extrinsic coagulation pathway...
September 6, 2017: Food & Function
Chalamaiah Meram, Jianping Wu
In this study, the egg yolk livetins (α, β, and γ-livetin) fraction and its hydrolysates, prepared by pepsin and Alcalase, were evaluated for their anti-inflammatory effects using lipopolysaccharide (LPS)-induced RAW 264.7 macrophages as an in vitro model. Enzymatic hydrolysis by pepsin and Alcalase successfully transformed the large molecular weight livetins into low molecular mass peptides mostly below 10kDa. Results revealed that livetins and its hydrolysates (peptides) treatment significantly reduced the inflammatory responses as evidenced by inhibition of production of nitric oxide (NO) (22...
October 2017: Food Research International
Juliane Floury, Tiago Bianchi, Jonathan Thévenot, Didier Dupont, Frédéric Jamme, Evelyne Lutton, Maud Panouillé, François Boué, Steven Le Feunteun
A novel time-lapse synchrotron deep-UV microscopy methodology was developed that made use of the natural tryptophan fluorescence of proteins. It enabled the monitoring in situ of the microstructural changes of protein gels during simulated gastric digestion. Two dairy gels with an identical composition, but differing by the coagulation mode, were submitted to static in vitro gastric digestion. The kinetics of gel particle breakdown were quantified by image analysis and physico-chemical analyses of digesta. The results confirm the tendency of rennet gels, but not acid gels, to form compact protein aggregates under acidic conditions of the stomach...
January 15, 2018: Food Chemistry
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