1,721,003 research outputs found
SERUM AMYLOID A IN RUMINANTS: DIAGNOSTIC VALUE AND FOOD CONTAMINATION ASSESSMENT
Full text linkThe aims of the work presented in this thesis were to investigate the bovine acute phase protein Serum Amyloid A, focussing on its value as safety marker in farm animals.
SAA can be considered as a natural anti-inflammatory and immunomodulatory agent and local expression of SAA, at the site of the initial acute phase reaction, could protect against the deleterious effects of inflammation.
In this study whether SAA can be isolated from tissues of bovine with clinical amyloidosis was investigated. We also investigated if AA fibrils present in milk can be then found in cheese after caseification, i.e. if the process of ripeining can degrade the AA fibrils.
In bovine, SAA was identified as potential marker of mastitis, and SAA milk concentration in milk increases before the raising of somatic cells.
In this thesis two aspect of the involvement of SAA in food safety were explored:
a)the acute phase reaction, strongly focused on the mammary gland. The animal model chosen was water buffalo, since no information is available so far about the acute phase reaction in this species. The acute phase proteins sequences are unknown, and also their concentration in physiological and pathological conditions are not established.
b)The possibility that high concentration of SAA in milk induce the formation of amyloid fibrils, which are considered to be potentially dangerous for human safety.
Results presented in this thesis advanced the knowledge of the acute phase reaction in water buffalo: the five APPs included in this investigation, namely Serum amyloid A, Haptoglobin, Ceruloplasmin, α1-acid glycoprotein and Lipolysaccaride Binding Protein were sequenced for the first time, and two of them were quantified.
In the second part of the thesis, we purified amyloid fibrils from amyloidosis-affected cows, and added purified fibrils at a given concentration in milk before ripening.
Results demonstrated the presence of insoluble fibrils in cheese added with amyloid proteins, even if a lower amount of precipitated insoluble SAA could be detected also in negative control cheese
In vitro effect of Interleukin-33 on gastric adenocarcinoma cell line (AGS) and intestinal epithelial cancer cell line (Caco-2)
No full textDichotomous effects of interleukin-33 on gastric and intestinal epithelial cell proliferation
No full textMesenchymal stem cells isolation from equine amnion and bone marrow for regenerative medicine applications
No full textDevelopment of a 3-D microstructurated scaffold as tracheal substitute : preliminary data
No full textAnalysis 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
Identification and characterization of Maternal Antigen That Embryos Require (Mater/NALP5) gene in pig somatic tissues and germ cells
No full textThe main objective of the present study was to evaluate the influence of pH and bicarbonate concentration in the activation or inhibition of European eel (Anguillaanguilla) spermatozoa and to evaluate the effect of different cryoprotectants: dimethyl sulphoxide (DMSO), acetamide, ethylene glycol, propanol, glycerol and methanol (MeOH). The effect of these factors was evaluated comparing the percentage of motile cells, the percentage of alive cells (by Hoechst staining) and the spermatozoa morphometry pre- and post-cryopreservation (by computer-assisted morphology analysis). Based on the above findings, three cryoprotectants (DMSO, MeOH and glycerol) were chosen and evaluated in two media (P1 and P1 modified) with different concentrations of NaHCO 3 and in the presence or absence of foetal bovine serum (FBS). The effect of these factors was evaluated comparing the percentage of alive and motile cells post-cryopreservation. DMSO was the cryoprotectant showing better results in relation to the percentage of spermatic alive cells post-freezing and caused a smaller modification of the head spermatozoa morphology. The combination of P1-modified medium with DMSO and containing FBS increased slightly but significantly the percentage of motile spermatozoa post-cryopreservation
Effect of pre-mating nutrition on mRNA levels of developmentally-relevant genes in sheep oocytes and granulosa cells
Full text linkThe present study was designed to investigate the relationship between pre-mating nutrition and the relative amounts of a panel of developmentally relevant genes in ovine oocytes and granulosa cells. Cast age ewes were fed a ration providing 0.5x (0.5 M) or 1.5x (1.5 M) live weight maintenance requirements for 2 weeks before slaughter. The ewes were synchronized and superovulated with FSH and pregnant mares serum gonadotropin. At slaughter, oocytes and granulosa cells were aspirated from follicles >2 mm in diameter and the relative abundance of 8 and 17 transcripts in oocytes and granulosa cells respectively were analyzed by semi-quantitative RT-PCR. In the oocytes, no differences between groups were observed for five transcripts (GDF9, BMP15, c-kit, glucose transporter 1 (SLC2A1), and hexokinase 1), but a lower amount of glucose transporter 3 (SLC2A3), sodium/glucose cotransporter 1 (SLC5A1), and Na+/K+ ATPase mRNAs was detected in the 0.5 M group. Increased expression of PTGS2, HAS2, and the leptin receptor long form was observed in granulosa cells from the 0.5 M group. No differences between groups were observed for the other transcripts (early growth response factor-1, estrogen receptor-, LH and FSH receptors, gremlin 1, pentraxin 3, KIT ligand, glucose transporters 1, 3, and 8, IGF1, IGF1 receptor, leptin receptor, and tumor necrosis factor-stimulated gene 6). Expression of leptin and sodium/glucose cotransporter 1 was not detected in both groups. The present data indicate that pre-mating nutrition is associated with alteration in the mRNA content in oocytes and surrounding follicle cells in ewes, which may account for the reduced reproductive performance typical of ewes that are fed a restricted ration for a short period of time before mating
Gene expression profile of ovine oocytes and granulosa cells with reference to premating nutrition
No full textPolimorfismi di sequenza del gene Mater/NALP5 e relazione genetica su razze bovine italiane
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