185 research outputs found
Proteomics in veterinary medicine: applications and trends in disease pathogenesis and diagnostics
No full textAdvancement in electrophoresis and mass spectrometry techniques along with the recent progresses in genomics, culminating in bovine and pig genome sequencing, widened the potential application of proteomics in the field of veterinary medicine. The aim of the present review is to provide an in-depth perspective about the application of proteomics to animal disease pathogenesis, as well as its utilization in veterinary diagnostics. After an overview on the various proteomic techniques that are currently applied to veterinary sciences, the article focuses on proteomic approaches to animal disease pathogenesis. Included as well are recent achievements in immunoproteomics (ie, the identifications through proteomic techniques of antigen involved in immune response) and histoproteomics (ie, the application of proteomics in tissue processed for immunohistochemistry). Finally, the article focuses on clinical proteomics (ie, the application of proteomics to the identification of new biomarkers of animal diseases)
Differential in-gel electrophoresis (DIGE) analysis of human bone marrow osteoprogenitor cell contact guidance
No full textWe have used a recent comparative proteomics technique, differential in-gel electrophoresis (DIGE), to study osteoprogenitor cell response to contact guidance in grooves. In order to increase protein output from small sample sizes, we used bioreactor culture before protein extraction and gel electrophoresis. Mass spectroscopy was used for protein identification. A number of distinct proteins were observed to exhibit significant changes in expression. These changes in protein expression suggest that the cells respond to tailored grooved topographies, with alterations in their proteome concurrent with changes in osteoprogenitor phenotyp
Effects of a surface topography composite with puerariae radix on human STRO-1-positive stem cells
No full textHuman skeletal stem cells (STRO-1 positive/STRO-1+) respond to different topographical features in various ways. On a flat surface these cells spread and tend to develop a fibroblast-like morphology. On a microgrooved surface enriched skeletal stem cell populations prefer to stretch along the grooves, which affects their cellular structure and differentiation, a phenomenon known as contact guidance. Growth factors, hormones and chemicals can also stimulate cell differentiation. A traditional Chinese medicine, puerariae radix, has previously been observed to stimulate bone formation. The active ingredients have been identified as isoflavones with estrogen-like bioactivity. This study combined the effects of microgrooved topology and hormone-like isoflavones in the biodegradable polymer polycaprolactone (PCL). Human osteogenic cells (STRO-1+) were cultured on flat PCL, grooved PCL and puerariae powder-impregnated grooved PCL for 5 weeks. Coomassie staining indicated that cell growth and survival was similar on flat PCL, grooved PCL and grooved PCL impregnated with 1 wt.% or 2 wt.% puerariae powder. Grooved PCL impregnated with 2 wt.% puerariae powder was observed to have an influence on protein expression, as observed by positive osteocalcin staining. Protein expression profiles were analyzed by difference gel electrophoresis to identify proteins that showed modulation of expression in response to these different environments. Overall, our results suggest that puerariae powder has an additive effect, along with microgrooved topographical stimulation, to promote changes in the STRO-1+ proteome that affect cell phenotype
Proteomics as a tool to explore human milk in health and disease
No full textProteins in milk have wide range of functions, they are carriers of minerals or chemically vulnerable and insoluble vitamins and other compounds, stabilisers of large aggregates or micelles of lipids, and components of both innate and acquired immune defence systems. Together with other components of milk, proteins may also contribute to the selection and establishment of appropriate microbiome in the gut of the infant. The proteome of mammalian milk is now known to be dynamic and changes radically with time after birth from colostrum to mature lactation. Significantly, immune and innate defence proteins appear in milk during infection of the mammary gland and possibly also during systemic infections. The understanding of the human milk proteome and how it changes with time during lactation and in disease is developing rapidly, and is to a large extent informed by proteomics of the milks of non-human mammals, domestic animals in particular. We review general methods now being applied for proteomic analysis of human milk. Moreover we place emphasis on how the milk proteome may change in different ways in response to disease, mastitis in particular, how such changes may be specific to pathogen types, and we give some insights about evolution
Tandem mass tag-based proteomics for studying the effects of low dietary ω6:ω3 fatty acid ratio during gestation and lactation on plasma protein profiles in pigs
No full textDifferential recognition patterns of Schistosoma haematobium adult worm antigens by the human antibodies IgA, IgE, IgG1 and IgG4
No full textAbstract Schistosoma haematobium antigen recognition profiles of the human isotypes IgA, IgE, IgG1 and IgG4 were compared by image analysis of western blots. Adult worm antigens separated by 2-dimensional gel electrophoresis were probed with pooled sera from Zimbabweans resident in a S. haematobium endemic area, followed by identification of individual antigenic parasite proteins using mass spectrometry. Overall, IgG1 reacted with the largest number of antigens, followed by IgE and IgA which detected the same number, while IgG4 detected the fewest antigens. IgE recognised all the antigens reactive with IgG4 as well as an additional four antigens; an isoform of 28kDaGST, phosphoglycerate kinase, actin 1 and calreticulin. IgG1 additionally recognised fatty acid binding protein, triose-phosphate isomerase, and heat shock protein 70, which were not recognised by IgA. Recognition patterns varied between some isoforms e.g. the 2 fructose 1-6-bis-phosphate aldolase isoforms differentially recognised by IgA and IgG1. Although the majority of S. haematobium adult worm antigens are recognised by all the four isotypes there are clear restrictions in antibody recognition for some antigens. This may partly explain differences observed in isotype dynamics at a population level. Differential recognition patterns for some isoforms indicated in the study have potential importance for vaccine developmen
Altered serum proteomes in newly weaned piglets born from the sows receiving a low ratio of n6:n3
No full textPerformance, oxidative status and serum proteome in piglets from sows fed at divergent ω6:ω3 ratios
No full textThis study aimed at assessing the effect of low versus high ratios of ω6:ω3 fatty acids in the gestation and lactation sow diets on their piglets. Sixteen multiparous sows were randomly allocated to two diets with ω-6:ω-3 ratios of 13:1 (C) and 4:1 (T) from d 28 of gestation onwards. Their post-weaning piglets were divided into four groups (10 piglets/group): C-without seaweed supplementation (C0); C-with seaweed (SW) at 4 g/kg feed (CSW); T-without seaweed (T0); and T-with seaweed (TSW). Body weight (BW), plasma and serum were collected at d 0, 7, 15 and 21 post-weaning. Pigs from CSW and T0 had higher (P<0.05) BW on d 15 and 21; average daily gain from d 0-21; feed intake from d 0-21; and gain:feed ratio from d 0-15. Protein and lipid oxidation products (AOPP and TBARS) in plasma were not different between groups. However, AOPP and total antioxidants (FRAP) decreased (P<0.0001) and TBARS increased (P<0.0001) from d 0 to 15. Moreover, FRAP was lower (P<0.05) in the SW group compared to the 0 group within the same sow diet on d 7 and FRAP tended to be lower (P=0.06) in piglets from T-mothers than those from C-mothers from d 0 to 21. A pilot proteomic experiment on serum showed some variations of the exponentially modified protein abundance index (emPAI) between C0 and CSW groups: albumin ranged from 280 to 604; haptoglobin from 0.47 to 8.16 whereas less difference was found on complement C3 (0.53-0.91) and alpha- 2-HS-glycoprotein (0.95-1.62). Further exploiting these results will aid to explain the mechanisms underlying the improved growth performance in piglets from sows receiving a low dietary ω6:ω3 ratio and fed SW post-weaning
Protein expression of STRO-1 cells in response to different topographic features
No full textHuman skeletal stem cells (STRO-1 positive) display distinct responses to different topographical features. On a flat surface, skeletal cells spread, and in vitro, they typically display a polarized, fibroblast-like morphology. However, on microgrooved surfaces, these cells prefer to stretch along the grooves forming a similar morphology to in vivo, bipolarized fibroblasts. In contrast, on nanopits, these cells display a polygonal and osteoblastic phenotype. We have examined mechanotransduction events of STRO-1 positive in response to fibroblastic, microgrooved and osteogenic, controlled disorder nanopit, topographies using proteomics after 3 days in culture. Protein expression profiles were analyzed by difference gel electrophoresis to identify proteins that showed modulation of expression in response to different topographic features to assess early decision events in these cells on these discrete topographies. After only 72 hours in culture, STRO-1 positive displayed differential regulations of families of proteins involved in cell migration and proliferation. The current study indicated that osteogenic decision specific events had already occurred. Runx2 was localized in nuclei of the skeletal stem cells on the osteogenic nanopits; however, few signaling pathway changes were observed. This study demonstrated that micro- and nanotopographies activated skeletal stem cells at different times and with distinct mechanotransduction profiles
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