In order to improve fish health and reduce use of chemotherapeutants in aquaculture production, the immunomodulatory effect of various nutritional ingredients has been explored. In salmon, there is ...evidence that functional feeds can reduce the abundance of sea lice. This study aimed to determine if there were consistent changes in the skin mucus proteome that could serve as a biomarker for dietary yeast cell wall extract. The effect of dietary yeast cell wall extract on the skin mucus proteome of Atlantic salmon was examined using two-dimensional gel electrophoresis. Forty-nine spots showed a statistically significant change in their normalised volumes between the control and yeast cell wall diets. Thirteen spots were successfully identified by peptide fragment fingerprinting and LC-MS/MS and these belonged to a variety of functions and pathways. To assess the validity of the results from the proteome approach, the gene expression of a selection of these proteins was studied in skin mRNA from two different independent feeding trials using yeast cell wall extracts. A calreticulin-like protein increased in abundance at both the protein and transcript level in response to dietary yeast cell wall extract. The calreticulin-like protein was identified as a possible biomarker for yeast-derived functional feeds since it showed the most consistent change in expression in both the mucus proteome and skin transcriptome. The discovery of such a biomarker is expected to quicken the pace of research in the application of yeast cell wall extracts.
Helicobacter pylori is a major human pathogen that infects the gastric mucosa and is responsible for a range of infections including gastritis and gastric carcinoma. Although other bacteria within ...the Helicobacter genus can also infect the gastric mucosa, there are Helicobacter species that infect alternative sites within the gastrointestinal (GI) tract. Two-dimensional gel electrophoresis was used to compare the cellular proteomes of seven non-pylori Helicobacters (H. mustelae, H. felis, H. cinaedi, H. hepaticus, H. fennelliae, H. bilis and H. cholecystus) against the more extensively characterised H. pylori. The different Helicobacter species showed distinctive 2D protein profiles, it was possible to combine them into a single dataset using Progenesis SameSpots software. Principal Component Analysis was used to search for correlations between the bacterial proteomes and their sites of infection. This approach clearly discriminated between gastric (i.e. those which infect in the gastric mucosa) and enterohepatic Helicobacter species (i.e. those bacteria that infect the small intestine and hepatobillary regions of the GI tract). Selected protein spots showing significant differences in abundance between these two groups of bacteria were identified by LC–MS. The data provide an initial insight into defining those features of the bacterial proteome that influence the sites of bacterial infection.
This study demonstrated that representative members of the Helicobacter genus were readily discriminated from each other on the basis of their in vitro whole cell proteomes determined using 2D gel electrophoresis. Despite the intra-species heterogeneity observed it was possible, to demonstrate that the enterohepatic (represented by H. bilis, H. hepaticus, H. fennelliae, H. cinaedi and H. cholecystus) and gastric (represented by H. pylori, H. mustelae, and H. felis) Helicobacters formed discrete groups based on their 2D protein profiles. A provisional proteomic signature was identified that correlated with the typical sites of colonisation of these members of the Helicobacter genus.
This article is part of a Special Issue entitled: Trends in Microbial Proteomics.
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•A proteomic comparison of non-pylori Helicobacter species was performed.•The data discriminated gastric and enterohepatic Helicobacters at the proteome level.•Proteins differentially synthesised between the two bacterial groups were identified.
Our knowledge of the physiology and health of small ruminants, specifically sheep and goats, is frequently obtained by extrapolating information from other species, for example the cow. However, ...there are important genetic, physiological and anatomical differences between small and large ruminants that cannot be ignored. This review considers the advances that have been made in the investigation of sheep and goat physiology through the use of proteomic technologies. Proteomics is widely used to analyze clinically relevant body fluids for a number of animals to define productive traits and health status biomarkers as well as to monitor therapeutic interventions for infectious and metabolic diseases. Although the proteomes of body fluids have been described in detail for some animal species, there are few equivalent studies for sheep and goats. Nevertheless, the data now available for the proteomes of a range of body fluids in small ruminants have helped define new diagnostic and prognostic markers for these species. Moreover, these data are beneficial in studies where these small ruminants serve as models for human disease. However, despite the progress achieved to date, comprehensive data on the specific proteomes for many tissues and body fluids for sheep and goats remain scarce. The aim of this review is to describe the current status of small ruminant proteomic research and to demonstrate the potential benefits, as well as highlight the difficulties, of working with these animals.
► Toxicity of methylmercury were assessed in kidney of Atlantic salmon (
salmo salar). ► Methylmercury accumulated in kidney and liver of the Atlantic salmon. ► Methylmercury affected proteomic ...markers related to oxidative stress, inflammation, and energy metabolism.
Methylmercury (MeHg) is an environmental contaminant most known for its severe neurotoxic effects. Although accumulation of MeHg tends to be several folds higher in kidney compared to other tissues, studies on nephrotoxic effects are almost non-existing. In this study we aim to investigate the toxicity of dietary MeHg in kidney of Atlantic salmon (
Salmo salar).
Atlantic salmon were exposed to dietary MeHg for a period of 8 weeks, before the fish were euthanized and kidney was sampled for proteomic and real time RT-PCR analysis, as well as for mercury determination. Protein separation was done with 2-D PAGE, and differentially regulated spots were picked for analysis using liquid chromatography MS/MS analysis. Moreover, whole blood and liver tissue were sampled for mercury determination and real time RT-PCR (liver).
MeHg exposed fish accumulated significantly more mercury (Hg) than control fish. The proteomic analysis revealed differential abundance of 26 spots in the kidney, and 14 of these protein spots were successfully identified. The proteins identified indicated effects of MeHg on; metabolism, inflammation, oxidative stress, protein-folding, and cell-structural components. Gene expression analysis of selected markers revealed few differentially regulated transcripts in kidney and liver in the exposed fish compared to the control fish. However, the affected transcripts indicated a disruption in the expression of two metabolic markers due to MeHg exposure in liver.
This study suggests that dietary MeHg has similar effects in kidney as previously shown for other tissues in fish. The effects observed were in markers for oxidative stress, inflammation and energy metabolism. The identification of proteomic markers in this study provides a basis for a better understanding of MeHg-induced nephrotoxicity in fish.
Bacterial infections are a major cause of morbidity and mortality throughout the world. By extending our understanding of the process of bacterial pathogenesis at the molecular level new strategies ...for their treatment and prevention can be developed. Proteomic technologies, along with other methods for global gene expression analysis, play an important role in understanding the mechanism(s) of bacterial pathogenesis. This review highlights the use of proteomics to identify protein biomarkers for virulent bacterial isolates and how these biomarkers can be correlated with the outcome of bacterial infection. Biomarker identification typically looks at the proteomes of bacteria grown under laboratory conditions. It is, however, the characterisation of the bacterial proteome during in vivo infection of its host that will eventually provide the most significant insights into bacterial pathogenesis. Although this area of research has significant technical challenges, a number of complementary proteome analytical approaches are being developed to identify and characterise the bacterial genes specifically expressed in vivo. Ultimately, the development of newly targeted therapies and vaccines using specific protein targets identified through proteomic analyses will be one of the major practical benefits arising from the proteomic analysis of bacterial pathogens.
Skeletal muscle is a complex tissue that is dominated by the presence of a few abundant proteins. This wide dynamic range can mask the presence of lower abundance proteins, which can be a confounding ...factor in large-scale proteomic experiments. In this study, we have investigated a number of pre-fractionation methods, at both the protein and peptide level, for the characterization of the skeletal muscle proteome. The analyses revealed that the use of OFFGEL isoelectric focusing yielded the largest number of protein identifications (>750) compared to alternative gel-based and protein equalization strategies. Further, OFFGEL led to a substantial enrichment of a different sub-population of the proteome. Filter-aided sample preparation (FASP), coupled to peptide-level OFFGEL provided more confidence in the results due to a substantial increase in the number of peptides assigned to each protein. The findings presented here support the use of a multiplexed approach to proteome characterization of skeletal muscle, which has a recognized imbalance in the dynamic range of its protein complement.
Urinary tract infections (UTIs) are among the most common of bacterial infections in humans. Although a number of Gram-negative bacteria can cause UTIs, most cases are due to infection by ...uropathogenic E. coli (UPEC). Genomic studies have shown that UPEC encode a number of specialized activities that allow the bacteria to initiate and maintain infections in the environment of the urinary tract. Proteomic analyses have complemented the genomic data and have documented differential patterns of protein synthesis for bacteria growing ex vivo in human urine or recovered directly from the urinary tracts of infected mice. These studies provide valuable insights into the molecular basis of UPEC pathogenesis and have aided the identification of putative vaccine targets. Despite the substantial progress that has been achieved, many future challenges remain in the application of proteomics to provide a comprehensive view of bacterial pathogenesis in both acute and chronic UTIs.
The ability to discriminate bacterial isolates is important for a number of areas of research in Medical Microbiology, particularly in defining bacterial taxonomy and monitoring transmission in ...epidemiological investigations. Molecular techniques capable of typing bacteria at the level of the genome and proteome are now widely used for these investigations. This review considers two electrophoretic methods for typing bacteria on the basis of their proteomes, namely 1-D SDS-PAGE and 2-DE. The application of these two techniques for bacterial typing is described with reference to two publications that appeared in Electrophoresis Costa, Electrophoresis 1990, 11, 382-391 and Cash et al., Electrophoresis 1997, 18, 1472-1482. Even though these methods have been used for nearly 20 years to differentiate bacterial isolates they remain key technologies in proteome-based typing methods. The developments that have arisen from the two key papers are described in order to highlight the advantages and disadvantages in typing bacteria at the level of their proteomes. Some of the difficulties associated with electrophoretic typing methods can be overcome by using non-gel proteomic methods based on MS to provide improved sensitivity and specificity. The application of proteomic methods to investigate bacterial taxonomy, epidemiology and pathogenesis in general has significant potential in furthering our understanding of infectious diseases.
The aim of this study is to provide a broad cultural description and analysis of Cayuse, Nez Perce, and Sahaptin language use. Investigative priority is given to the behavioral correlates of fluent ...and semi-fluent speaker choices and the discursive consciousness that informs them. The findings show how language use is organized and embodied as ‘ways of speaking’ in traditional cultures of the Indigenous Southern Plateau region both as a responsive system to societal change and as a semiotic behavioral resourse for cultural continuity. The motivation for this study arises from my belonging to the Cayuse, Nez Perce, and Sahaptin speech communities where this research was conducted as well as from the growing global awareness concerning the endangered status of Indigenous languages in the Indigenous Southern Plateau and elsewhere throughout the world. It is hoped that the findings and language data contained in this language documentation research can inform and contribute to positive outcomes centering in the revitalization of culture and language in the Indigenous Southern Plateau.
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