Laura Restelli

Escherichia coli lipopolysaccharides and Staphylococcus aureus enterotoxin B differentially modulate inflammatory microRNAs in bovine monocytes.

MicroRNAs (miRNAs) are a family of regulatory molecules involved in many physiological processes, including activation of cells of the immune system. This study investigated the effect of Escherichia coli lipopolysaccharide (LPS) and Staphylococcus aureus enterotoxin B (SEB) on the expression of five miRNAs involved in the inflammatory response, including miR-9, miR-125 b, miR-155, miR-146 a and miR-223, in bovine CD14(+) cells (monocytes). Incubation of monocytes with SEB induced down-regulation of miR-155, miR-223 and miR-125 b, but not the anti-inflammatory miRNA miR-146 a. Conversely, incubation with LPS upregulated both miR-155 and miR-146 a. In vitro incubation of isolated CD14(+) bovine monocytes with LPS and SEB elicited different and opposite expression of miRNAs reportedly involved in inflammatory reactions.

Proteomics in farm animals models of human diseases

The need to provide in vivo complex environments to understand human diseases strongly relies on the use of animal models, which traditionally include small rodents and rabbits. It is becoming increasingly evident that the few species utilised to date cannot be regarded as universal. There is a great need for new animal species that are naturally endowed with specific features relevant to human diseases. Farm animals, including pigs, cows, sheep and horses, represent a valid alternative to commonly utilised rodent models. There is an ample scope for the application of proteomic techniques in farm animals, and the establishment of several proteomic maps of plasma and tissue has clearly demonstrated that farm animals provide a disease environment that closely resembles that of human diseases. The present review offers a snapshot of how proteomic techniques have been applied to farm animals to improve their use as biomedical models. Focus will be on specific topics of biomedical research in which farm animal models have been characterised through the application of proteomic techniques.

LC-MS/MS analysis of visceral and subcutaneous adipose tissue proteomes in young goats with focus on innate immunity and inflammation related proteins

UNLABELLED The endocrine role of adipose tissue and its involvement in several physiological and pathological processes are well recognized. Studies on human, mouse and rat adipose tissues have made clear that subcutaneous and visceral deposits play different roles, which is also reflected by different protein and gene expression patterns. In ruminants, fat tissues play important biological roles not only for animal health, but also for quality and gain in meat and milk production. Yet very few studies have explored the ruminant adipose tissue proteomes. The aim of our study was to compare subcutaneous and visceral adipose tissues of goat, focusing on proteins involved in immune and inflammatory response. A 2-D LC-MS/MS approach followed by cluster analysis shows a clear distinction between subcutaneous and visceral fat tissue proteomes, and qualitative RT-PCR based analysis of 30 potential adipokines further confirmed the individual expression patterns of 26 of these, including 7 whose mRNA expression was observed for the first time in adipose tissues. This study provides a first description of adipose tissue proteomes in goat, and presents observations on novel proteins related to metabolic and inflammatory pathways. The mass spectrometry data have been deposited to the ProteomeXchange Consortium with the data set identifier PXD000564. BIOLOGICAL SIGNIFICANCE The proteomic analysis of different subcutaneous and visceral adipose tissue deposits showed tissue specific differences in protein expressions of well known as well as novel adipokines. This highlights the importance of sampling site when studying adipose tissues metabolic roles. The protein expression characteristics of adipose tissues was evaluated by quantitative RT-PCR, and confirmed that adipose tissues play a central role in controlling inflammation, detoxification and coagulation pathways, as well as regulation of body fat mobilization in dairy animals. These findings are of particular interest in farm animals where health and production traits are important for animal welfare and for economic gains.

Expression of α1-acid glycoprotein and lipopolysaccharide binding protein in visceral and subcutaneous adipose tissue of dairy cattle

Adipose tissue is an endocrine compartment that plays an important role in immune defence by producing and releasing a wide range of proteins, including acute phase proteins (APPs). The liver is the main organ of APP synthesis, although extrahepatic production has also been reported. In the present study, expression of two APPs in dairy cattle, lipopolysaccharide binding protein (LBP) and α1-acid glycoprotein (AGP), was determined in four visceral (pericardial, mesenteric, omental and retroperitoneal) and three subcutaneous (withers, tail head and sternum) adipose tissue depots. mRNA expression was evaluated using qualitative and quantitative PCR, protein profiles were assessed by Western blot analysis and cellular localisation was determined by immunohistochemistry. The presence of LBP and AGP was demonstrated at mRNA and protein levels in all seven adipose tissue depots. Expression of AGP and LBP suggests that they may have roles as local and systemic inflammatory adipokines.

Proteomics and protein analyses of ovine and caprine body fluids: current studies and future promises.

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.

Hyposialylated α1-acid glycoprotein inhibits phagocytosis of feline neutrophils

Abstract Feline α1-acid glycoprotein (fAGP) modifies both its serum concentration and its glycan moiety during diseases. fAGP is hyposialylated in cats with feline infectious peritonitis (FIP), but not in clinically healthy cats or in cats with other diseases. This study was aimed to determine whether hyposialylated fAGP influences phagocytosis. A flow cytometric method based on ingestion of fluoresceinated bacteria and adapted to feline blood was used to assess phagocytosis of leukocytes incubated with ‘non-pathological’ fAGP (purified from sera with normal concentrations of AGP) and ‘pathological’ fAGP (purified from sera with >1.5mg/mL hyposialylated AGP). The flow cytometric method provided repeatable results for neutrophils (coefficients of variations, CVs <15%) but not for monocytes (CVs>20%) which had also a high individual variability. Compared with saline solution and with non-pathological fAGP, pathological fAGP significantly decreased phagocytosis in neutrophils and monocytes. This study demonstrated that hyposialylated fAGP down-regulates the phagocytic activity of feline neutrophils.

Protein expression in bovine mononuclear cells after stimulation with lipopolysaccharides and lipoteichoic acid: a proteomic approach

The aim of this project was to investigate the proteome of bovine mononuclear cells in physiological condition and after stimulation with different pathogen associated molecular patterns.

Bitter taste in water-buffalo (Bubalus bubalis): from T2R gene identification to expression studies

Among farm animals the water buffalo (Bubalus bubalis) is largely used for milk, meat, and fertilizer production, representing an important fraction for the economy of many countries, especially in Tropical and Mediterranean areas. Taste plays a crucial role for animal regulation of food intake, being a determinant factor for them to chose suitable and reject unsuitable foods, hence defining their ingestive behaviour. Being bitter is considered the most interesting taste modality in herbivores due to its evolution to prevent the consumption of plant toxins. It is a very important factor to consider in animal production, as it is highly related to food preferences. In ruminants, the knowledge of genes and molecular mechanisms behind bitter taste perception is scarce and most of the studies were carried out only in cattle, due to the lack of genomic data for the other species.

LC-MS/MS analyses of visceral and subcutaneous adipose tissue proteomes in goats

Adipose tissue (AT) is considered a group of highly active endocrine organs, taking part in regulating several metabolic processes, including reproduction, inflammatory response, and production and secretion of signalling molecules with important biological roles, also known as adipokines. Different deposits of visceral and subcutaneous origin differ in mRNA abundance of several adipokines in sheep (Lemor et al., 2010); few studies have been carried out at a proteomic level in ruminant adipose tissue (Rajesh et al., 2012) and none in goats.

Proteomic analysis of visceral and subcutaneous adipose tissue in goats

Adipose tissue (AT) has for a long time been regarded as a simple storage tissue for energy in the form of triglycerides. There is currently accumulating scientific evidence that adipose tissue is a highly active organ, that actively takes part in regulating several metabolic processes, including reproduction, inflammatory response, and production and secretion of signalling molecules that have important biological roles, also known as adipokines. These most importantly include tumor necrosis factor alpha (TNF-alpha), interleukins, leptin and adiponectin.