Proteomic evaluation of sheep serum proteins.

BACKGROUND: The applications of proteomic strategies to ovine medicine remain limited. The definition of serum proteome may be a good tool to identify useful protein biomarkers for recognising sub-clinical conditions and overt disease in sheep. Findings from bovine species are often directly translated for use in ovine medicine. In order to characterize normal protein patterns and improve knowledge of molecular species-specific characteristics, we generated a two-dimensional reference map of sheep serum. The possible application of this approach was tested by analysing serum protein patterns in ewes with mild broncho-pulmonary disease, which is very common in sheep and in the peripartum period which is a stressful time, with a high incidence of infectious and parasitic diseases. RESULTS: This study generated the first reference 2-DE maps of sheep serum. Overall, 250 protein spots were analyzed, and 138 identified.Compared with healthy sheep, serum protein profiles of animals with rhino-tracheo-bronchitis showed a significant decrease in protein spots identified as transthyretin, apolipoprotein A1 and a significant increase in spots identified as haptoglobin, endopin 1b and alpha1B glycoprotein.In the peripartum period, haptoglobin, alpha-1-acid glycoprotein, apolipoprotein A1 levels rose, while transthyretin content dropped. CONCLUSIONS: This study describes applications of proteomics in putative biomarker discovery for early diagnosis as well as for monitoring the physiological and metabolic situations critical for ovine welfare.

D-galactose/D-glucose-binding Protein from Escherichia coli as Probe for a Non-consuming Glucose Implantable Fluorescence Biosensor.

D-Galactose/D-glucose-binding protein from E. coli (GGBP) is a monomer thatbinds glucose with high affinity. The protein structure of GGBP is organized in twoprincipal domains linked by a hinge region that form the sugar-binding site. In this workwe show that the mutant form of GGBP at the amino acid position 182 can be utilized as aprobe for the development of a non-consuming analyte fluorescence biosensor to monitorthe glucose level in diabetes health care.

Ovine subclinical mastitis: proteomic analysis of whey and milk fat globules unveils putative diagnostic biomarkers in milk.

Subclinical mastitis is one of the main causes of alteration in milk content and has a major impact on both animal welfare and economy in the dairy industry. A better knowledge is needed to understand the ovine mammary gland metabolism and its response to bacterial infection. In this study, the proteomic changes in ovine milk as a result of subclinical mastitis were investigated by comparing both whey and fat globule membrane profiles of samples from Staphylococcus chromogenes-positive individuals, with those from non-infected counterparts having high or low somatic cell count; the latter were used as control. 2-DE and combined MS procedures were utilized for this purpose. Although sample bromatological parameters were very similar, proteomic analysis highlighted significant differences between the three experimental groups. Most relevant changes were observed between samples of infected milk and control. Modifications related to the defense response of the mammary gland to the pathogen were evident, with important consequences on nutritional and technological properties of milk. On the other hand, quantitative protein changes between non-infected samples with low and high levels of somatic cells indicated that the latter may result as a consequence of a probable unpaired cellular metabolism due to cellular stress, hormonal variations or previous infections. Putative biomarkers useful for the monitoring of sheep mammary metabolism and for the careful management of ovine subclinical mastitis to avoid its clinical degeneration are proposed and discussed. BIOLOGICAL SIGNIFICANCE: Proteomics has been here applied to the differentiation of healthy and subclinical mastitic sheep milk samples, evidencing the response of the mammary gland to S. chromogenes infection. Presented results propose useful protein biomarkers for the detection of ewe mammary infection at its subclinical stages and, subsequently, mastitis recognition and treatment. Differently from bovine, these data confirm that the increase in somatic cell count in sheep milk is not always associated with protein factors that characterize the mammary gland infection; accordingly, somatic cell count cannot be considered as a useful parameter to certainly diagnose subclinical mastitis in ovine.

Plasma protein changes in horse after prolonged physical exercise: a proteomic study.

Physical exercise induces various stress responses and metabolic adaptations that have not yet been completely elucidated. Novel biomarkers are needed in sport veterinary medicine to monitor training levels and to detect subclinical conditions that can develop into exercise-related diseases. In this study, protein modifications in horse plasma induced by prolonged, aerobic physical exercise were investigated by using a proteomic approach based on 2-DE and combined mass spectrometry procedures. Thirty-eight protein spots, associated with expression products of 13 genes, showed significant quantitative changes; spots identified as membrane Cu amine oxidase, α-1 antitrypsin, α-1 antitrypsin-related protein, caeruloplasmin, α-2 macroglobulin and complement factor C4 were augmented in relative abundance after the race, while haptoglobin ß chain, apolipoprotein A-I, transthyretin, retinol binding protein 4, fibrinogen γ chain, complement factor B and albumin fragments were reduced. These results indicate that prolonged physical exercise affects plasma proteins involved in pathways related to inflammation, coagulation, immune modulation, oxidant/antioxidant activity and cellular and vascular damage, with consequent effects on whole horse metabolism.

Gambling on putative biomarkers of osteoarthritis and osteochondrosis by equine synovial fluid proteomics.

Osteoarthritis (OA) and osteochondrosis (OC) are two of the main challenges in orthopedics, whose definitive diagnosis is usually based on radiographic/arthroscopic evidences. Their early diagnosis should allow preventive or timely therapeutic actions, which are generally precluded from the poor relationships occurring between symptomatologic and radiographic evidences. These limitations should be overcome by improving the knowledge on articular tissue metabolism and on molecular factors regulating its normal homeostasis, also identifying novel OA and OC biomarkers suitable for their earlier diagnoses, whenever clinical/pathological inflammatory scenarios between these joint diseases seem somewhat related. To identify proteins involved in their aetiology and progression, we undertook a differential proteomic analysis of equine synovial fluid (SF), which compared the protein pattern of OA or OC patients with that of healthy individuals. Deregulated proteins in OA and OC included components related to inflammatory state, coagulation pathways, oxidative stress and matrix damage, which were suggestive of pathological alterations in articular homeostasis, plasma-SF exchange, joint nutritional status and vessel permeability. Some proteins seemed commonly deregulated in both pathologies indicating that, regardless of the stimulus, common pathways are affected and/or the animal joint uses the same molecular mechanisms to restore its homeostasis. On the other hand, the increased number of deregulated proteins observed in OA with respect to OC, together with their nature, confirmed the high inflammatory character of this disease. Some deregulated proteins in OA found a verification by analyzing the SF of injured arthritic joints following autologous conditioned serum treatment, an emergent therapy that provides positive results for both human and equine OA. Being the horse involved in occupational/sporting activities and considered as an excellent animal model for human joint diseases, our data provide suggestive information for tentative biomedical extrapolations, allowing to overcome the limitations in joint size and workload that are typical of other small animal models.