1,721,077 research outputs found
Proteomics in farm animals models of human diseases
No full textThe 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
The acute phase protein α1-acid glycoprotein : a model for altered glycosylation during diseases. : Updates and new perspectives
No full textWidespread expression of SAA and Hp RNA in bovine tissues after evaluation of suitable reference genes
No full textThe serum amyloid A (SAA) and haptoglobin (Hp) are the most prominent acute phase proteins (APPs) in cow. Liver mainly produces APPs, but extra hepatic expression has also been demonstrated in some tissues. The major aim of the present study was to assess the constitutive SAA and Hp mRNA expression by quantitative PCR (qPCR) in a wide panel of 33 bovine tissues, including gastrointestinal tract, respiratory system, urogenital system, mammary gland, hematopoietic system, central nervous system, eye, thyroid and heart. Normalization of gene expression in different samples requires reference genes, which are stably expressed. Therefore, seven reference genes were investigated (ACTB, GAPDH, HMBS, SDHA, YWHAZ, SF3A1, EEF1A2) and three genes, namely SF3A1, HMBS and ACTB, were selected after assessing their stability with geNorm and NormFinder() softwares. The qPCR analysis confirmed liver as the principal source of SAA and Hp, but also identified both APPs' mRNA in almost all tissues. The highest expression rate of SAA was found in thyroid, followed by pancreas and submandibulary gland. Hp mRNA expression was detected at high concentration in pancreas and submandibulary gland. The present data indicated a widespread expression of SAA and Hp also in non pathological conditions, thus envisaging a possible role as immunomodulatory and protective molecules. To understand where SAA and Hp come from is the prerequisite to their utilization as Acute Phase Reaction biomarkers
Proteomics and metabolomics characterizing the pathophysiology of adaptive reactions to the metabolic challenges during the transition from late pregnancy to early lactation in dairy cows
Full text linkThe transition from late pregnancy to early lactation is a critical period in a dairy cow's life due to the rapidly increasing drain of nutrients from the maternal organism towards the foetus and into colostrum and milk. In order to cope with the challenges of parturition and lactation, comprehensive adaptive reactions comprising the endocrine and the immune system need to be accomplished. There is high variation in this coping ability and both metabolic and infectious diseases, summarized as âproduction diseasesâ, such as hypocalcaemia (milk fever), fatty liver syndrome, laminitis and ketosis, may occur and impact welfare, productive lifespan and economic outcomes. Proteomics and metabolomics have emerged as valuable techniques to characterize proteins and metabolite assets from tissue and biological fluids, such as milk, blood and urine. In this review we provide an overview on metabolic status and physiological changes during the transition period and the related production diseases in dairy cows, and summarize the state of art on proteomics and metabolomics of biological fluids and tissues involved in metabolic stress during the peripartum period. We also provide a current and prospective view of the application of the recent achievements generated by omics for biomarker discovery and their potential in diagnosis. Biological significance: For high-yielding dairy cows there are several âoccupational diseasesâ that occur mainly during the metabolic challenges related to the transition from pregnancy to lactation. Such diseases and their sequelae form a major concern for dairy production, and often lead to early culling of animals. Beside the economical perspective, metabolic stress may severely influence animal welfare. There is a multitude of studies about the metabolic backgrounds of such so called production diseases like ketosis, fatty liver, or hypocalcaemia, although the investigations aiming to assess the complexity of the pathophysiological reactions are largely focused on gene expression, i.e. transcriptomics. For extending the knowledge towards the proteome and the metabolome, the respective technologies are of increasing importance and can provide an overall view of how dairy cows react to metabolic stress, which is needed for an in-depth understanding of the molecular mechanisms of the related diseases. We herein review the current findings from studies applying proteomics and metabolomics to transition-related diseases, including fatty liver, ketosis, endometritis, hypocalcaemia and laminitis. For each disease, a brief overview of the up to date knowledge about its pathogenesis is provided, followed by an insight into the most recent achievements on the proteome and metabolome of tissues and biological fluids, such as blood serum and urine, highlighting potential biomarkers. We believe that this review would help readers to be become more familiar with the recent progresses of molecular background of transition-related diseases thus encouraging research in this field
Analysis of bovine alpha1-acid glycoprotein phosphorylation by 2-D electrophoresis and IN-GEL fluorescence
No full textAlpha1-acid glycoprotein is considered an acute phase proteins (APP) in most species. One of the most interesting features of AGP is that its concentration in plasma not only rises during inflammation, but also undergoes structural modifications of its oligosaccharide moiety (more than 40% of the molecular mass of the protein), resulting in a change of both the degree of branching and fucosylation. Furthermore, a throughout analysis of the primary structure revealed that AGP’s sequence presents also seven potential phosphorylation sites. No studies report the actual phosphorylation of the protein.
AGP purified from bovine serum was submitted to 2D-electrophoresis and the separates isoforms of the protein has been analyzed by mean of a general fluorescence dye and Phos-tag phosphoprotein gel stain dye, which selectively binds the phophoserine, phosphothreonine and phosphotyrosine residues. The binding has been detected by in-gel fluorescence.
The 2D maps clearly reveal the purified AGP from healthy animals presents a marked microheterogeneity, concerning both the pI and MW. At least three different families of protein isoforms can be detected: the first is heterogeneous in charge but is not phosphorylated, the second is heterogeneous both in charge and MW, indicating also a different glycosylation pattern, and the third is heterogeneous in charge and appear to be strongly phosphorylated.
In conclusion, the observed electrophoretic charge microheterogeneity of AGP may be due to the phosphorylation status of the protein and, possibly, to different levels of amidation of the glutammic and aspartic acid residues.
Although the findings predented in this communication should be considered as still preliminary, they show that, further to gene expression control and glycosylation status, the activity of AGP may be also regulated by phosphorylation events. The next step of this investigation will be to evaluate if phosphorylation of AGP may be modified and regulated during diseases
Real-time pcr per la quantificazione dell'espressione del recettore tir8 sigirr in tessuti di bovino
No full textVALUTAZIONE DI PROTEINE DI FASE ACUTA NEL PERIPARTO DELLA CAPRA
No full textHaptoglobin (Hp), Serum Amyloid A (SAA), Alpha 1 acid glycoprotein (AGP)
and Ceruloplasmin were evaluated in blood serum from 24 goats, starting from 2 days before
parturition until 7 days after. Metabolic profile was also assayed. 17 goats were clinically
healthy, 4 presented dystocia, 2 retained placenta and 1 bronchopneumonia. In healthy goats Hp
increased significantly at 1 and 2 days after parturition; SAA increased in similar way, but
without statistical significance. Goats with dystocia showed higher SAA values after delivery.
The goat with bronchopneumonia had high level of AGP. All but one goats carried two or three
fetuses; NEFA and B-OH butyrate were elevated at parturition and decreased significantly from
the day after; no positive correlation were observed with AP
Acute phase protein response in Alpine ibex with sarcoptic mange
No full textThe acute phase proteins (APP) are a group of serum proteins that change their concentration in animals following external or internal challenges, such as infection, inflammation or stress. The concentrations of four APPs, including serum amyloid A (SAA), haptoglobin (Hp), α1-acid glycoprotein (AGP) and ceruloplasmin (Cp) were determined in serum collected from healthy Alpine ibexes (Capra ibex) and ibexes with Sarcoptes scabiei mange. Primary structures of all four APPs were determined by cDNA sequencing. The concentrations of all four APPs were higher in serum of animals with clinical signs of sarcoptic mange when compared to healthy animals. Two of the APPs, including SAA and AGP, acted as major APPs, since their serum concentrations were increased more than 10-folds when compared to healthy animals (P < 0.001). The other two APPs, including Hp and Cp, acted as minor acute phase proteins, as their concentrations were increased from two to five folds (P < 0.001).
These findings provide a remarkable potential as diagnostic markers for the early detection of sarcoptic mange in free ranging animals
Extra hepatic expression of the acute phase protein alpha 1-acid glycoprotein in normal bovine tissues
No full textCombined quantitative (real-time) polymerase chain reaction and immunohistochemistry were used to evaluate the expression of the minor acute phase protein alpha 1-acid glycoprotein (AGP, orosomucoid) in bovine extrahepatic tissues. AGP was produced mainly in the salivary glands and spleen, whereas minor expression was detected in all other tissues sampled, including lung, lymph nodes, uterus, ovary, kidney and tongue. The findings were consistent with immunohistochemical results. In view of the immunomodulatory and direct antibacterial activity of AGP, its expression in the salivary glands may signal an involvement in the regulation of the local immunity, even in non-pathological conditions
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