An analytical approach using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) separated proteins from the brain tissue of the fish Paralichthys olivaceus. Approximately 600 protein spots were detected from the brain sample when applying 600 microg protein to a 2D-PAGE gel in the pH range 3.5-10.0. Compared to a control sample, significant changes of 24 protein spots were observed in the fish tissue exposed to acute toxicity of seawater cadmium (SCAT) at 10 ppm for 24h. Among these spots, nine were down-regulated, nine were up-regulated, two showed high expression, and four showed low expression. The collected spots were identified by peptide mass fingerprinting (PMF) and database search, and they were further classified by LOC tree, a hierarchical system of support vector machines which predict their sub-cellular localization. The amount of transferrin expression in brain cells decreased linearly with the increase of SCAT concentration in seawater. Among the 24 proteins identified on a 2D-PAGE gel, 9 demonstrated a synchronous response to acute cadmium, suggesting that they might represent a biomarker profile. Based on their variable levels and trends on the 2D-PAGE gel this protein (likely to be transferrin) suggesting they might be utilized as biomarkers to investigate cadmium pollution levels in seawater and halobios survival, as well as to evaluate the degree of risk of human fatalities. The results indicate that the application of multiple biomarkers has an advantage over a single biomarker for monitoring levels of environmental contamination.
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http://dx.doi.org/10.1016/j.aquatox.2006.02.010 | DOI Listing |
Bio Protoc
December 2024
Key Laboratory of Sports and Physical Fitness of the Ministry of Education, Beijing Sport University, Beijing, China.
Proteomics analysis is crucial for understanding the molecular mechanisms underlying muscle adaptations to different types of exercise, such as concentric and eccentric training. Traditional methods like two-dimensional gel electrophoresis and standard mass spectrometry have been used to analyze muscle protein content and modifications. This protocol details the preparation of muscle samples for proteomics analysis using ultra-high-performance liquid chromatography (UHPLC).
View Article and Find Full Text PDFVet Parasitol
December 2024
Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China. Electronic address:
Currently, the 'gold standard' for diagnosis of Psoroptes ovis infections is detecting Psoroptes mites or eggs in skin scrapings under microscopy, but it is prone to be mis-diagnosed for detecting early infection of P. ovis. Hence, seeking a reliable diagnostic technique for detecting early-stage mite infections is extremely desirable.
View Article and Find Full Text PDFFront Cell Dev Biol
December 2024
Proteomics, Lipidomics and Metabolomics Core Facility, School of Life Sciences, Faculty of Science, University of Technology Sydney, Ultimo, NSW, Australia.
Introduction: The identification of effective, selective biomarkers and therapeutics is dependent on truly deep, comprehensive analysis of proteomes at the proteoform level.
Methods: Bovine serum albumin (BSA) isolated by two different protocols, cold ethanol fractionation and heat shock fractionation, was resolved and identified using Integrative Top-down Proteomics, the tight coupling of two-dimensional gel electrophoresis (2DE) with liquid chromatography and tandem mass spectrometry (LC-MS/MS).
Results And Discussion: Numerous proteoforms were identified in both "purified" samples, across a broad range of isoelectric points and molecular weights.
A previously developed algorithm for the preliminary identification of protein proteoforms associated with post-translational modifications (PTMs) based on 2D electrophoresis data (DOI: 10.18097/BMCRM00191) has been used in this study for analysis of experimental data obtained using mice and reported in two papers by different authors. The authors of the first paper identified 8 groups of spots on 2D electrophoretic maps corresponding to 8 proteins with at least two unconcretised proteoforms.
View Article and Find Full Text PDFChemosphere
December 2024
BRIC-National Institute of Animal Biotechnology (NIAB), Hyderabad, 500032, Telangana, India; BRIC- Regional Centre for Biotechnology (RCB), Faridabad, 121001, Haryana, India. Electronic address:
Endosulfan (Ed), a widely used organochlorine pesticide, is classified as a persistent organic pollutant (POP). Its long half-life, resistance to degradation, and bioaccumulation in the food chain contaminates soil, water, and air. Such widespread environmental damage triggers monitoring its levels for ensuring compliance with safety regulations and protecting public health.
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