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Biomedical subjects

F Rosi

Publications and source records attributed to F Rosi.

At least 19 recordsLinked to original sources

Leptin in bovine colostrum and milk.

We studied leptin content in bovine colostrum, milk and plasma during the first month of lactation, and investigated relationships between selected milk components and milk leptin in five multiparous dairy cows. Colostrum/milk yield and composition were measured on days 0, 10, 20, and 30 of lactation. Leptin was assayed using a multi-species leptin RIA kit. Leptin concentration was 56 % lower in mature milk (day 10) than colostrum (13.90 vs. 6.14 microg/l; p < 0.001), but remained steady over the twenty days afterwards. Daily secretion of leptin into mature milk was 28 % lower than into colostrum (173.2 microg/d vs. 220.0 microg/d; p = 0.09) notwithstanding an 80 % increase in production. Colostrum and milk leptin levels correlated with fat (0.90; p < 0.001) and choline phospholipid (0.76; p < 0.05). Plasma and milk leptin decreased during the first month, but remained higher in milk, and highest in colostrum. Thus, leptin is present in large quantities in colostrum, less so and more variably in untreated milk, and is likely to be decreased in skimmed milk. These findings have implications for the use of untreated milk and colostrum-based (functional) food products.

Animals↗

Purine metabolism in B-cell lymphocytic leukemia: a microarray approach.

B-cell chronic lymphocytic leukemia (B-CLL) is an adult-onset highly heterogeneous malignancy characterized by a cells resistance to apoptosis rather than to highly proliferative cells. In previous research, we evidenced an imbalance of purine metabolism in B-CLL cells. Since the extracellular adenosine has been proved to induce apoptosis via A2b receptor, enzymes involved in adenosine metabolism could play an important role in apoptosis resistance of B-CLL cells. We prepared a microarray chip for the analysis of 50 selected genes that could be of interest in B-CLL: enzymes of purine de-novo, salvage and catabolic pathway, oxidative stress enzymes, and apoptotis-related proteins. Preliminary results identify many genes of purine metabolism that exhibit low or high expression, while genes involved in signal transduction and apoptosis exhibit lower alterations even if of remarkable interest. This application of microarray technique seems promising and at least a subset of these genes will be valid candidates for further studies.

Adenosine↗

Liver transplant: adenosine metabolism and apoptosis.

Apoptosis and necrosis coexist in ischemia-reperfusion (I/R) injury following organ transplant. During experimental liver transplant we evidenced a deep alteration in energy and antioxidant status. The activity of purine catabolic enzymes was also altered. Caspase-3 (C-3), protein tyrosine phosphatase (PTP) showed significative alterations that lead to DNA fragmentation. These findings could be of interest in new potential strategy to prevent and treat I/R injury.

Adenosine↗

Metabolism of adenosine in human colorectal tumour.

The aim of this work is to analyse the activities of the enzymes metabolising adenosine in fragments of neoplastic and normal-appearing mucosa, surrounding the tumour in 20 patients affected by colorectal cancer. The results show that the activities of the enzymes are markedly higher in tumour in comparison to normal mucosa to coope with the accelerated purine metabolism in cancerous tissues.

Adenosine↗

Enzyme activities controlling adenosine levels in normal and neoplastic tissues.

Adenosine is known to be associated with effects such as inhibition of immune response, coronary vasodilation, stimulation of angiogenesis, and inhibition of inflammatory reactions. Some authors suggest that adenosine may also have similar functions in tumor tissues. Tissue levels of adenosine are under close regulation by different enzymes acting at different levels. Adenosine is produced from AMP by the action of 5'-nucleotidase (5'-NT) and is converted back into AMP by adenosine kinase (AK) or into inosine by adenosine deaminase (ADA). Inosine is converted into purine catabolites by purine nucleoside phosphorylase (PNP), whereas AMP is converted into ADP and ATP by adenylate kinase (MK). The aim of this study was to analyze the activities of the above enzymes in fragments of neoplastic and apparently normal mucosa, obtained less than 5 cm and at least 10 cm from tumors, in 40 patients with colorectal cancer. The results showed much higher activities of ADA, AK, 5'-NT, and PNP in tumor tissue than in neighboring mucosa (p > 0.01 for ADA, AK, and PNP; p > 0.05 for 5'-NT), suggesting that the activities of purine metabolizing enzymes increase to cope with accelerated purine metabolism in cancerous tissue. The simultaneous increase in ADA and 5'-NT activities might be a physiological attempt by cancer cells to provide more substrate to accelerate salvage pathway activity.

Adenosine↗

Cardiac surgery: myocardial energy balance, antioxidant status and endothelial function after ischemia-reperfusion.

Myocardial and endothelial damage is still a widely debated problem during the ischemia-reperfusion sequence in heart surgery. We evaluated myocardial purine metabolites, antioxidant defense mechanisms, oxidative status and endothelial dysfunction markers in 14 patients undergoing coronary artery by-pass graft (CABG). Heart biopsies were taken before aortic cross-clamping (t1), before clamp removal (t2) and 30 min after reperfusion (t3); perchloric extracts of the tissue were analyzed for glutathione, NAD, nucleotide nucleoside and base content by capillary electrophoresis (CE). In plasma samples from the coronary sinus we evaluated: nitrate and nitrite concentrations by CE, plasma glutathione peroxidase (plGPx) by ELISA, endothelin-1 (ET-1) by RIA and reactive oxygen metabolites (ROM) by colorimetric assay. During the ischemic period (t2) we observed a reduction in cellular NAD and GSH levels, as well as nitrate, nitrite and plGPx. ATP and GTP levels decreased and their catabolic products AMP, GMP, IMP, adenosine, inosine and hypoxanthine accumulated. The energy charge, ATP/ADP ratio, and nucleotide/(nucleoside + base) ratios decreased. At t3, levels of plasma ET-1 increased and monophosphate nucleotides tended to return to basal values. The energy charge did not increase but the nucleotide/(nucleoside + nucleobase) ratio recovered to some extent. Levels of nitrates plus nitrites continued to decrease. No significant variation in ROM levels was observed. Our data indicate that oxidative stress and endothelial damage are major events during CABG, overwhelming the scavenging capacity of the myocyte and preventing restoration of the normal energy balance for 30 min after reperfusion. The AMP deaminase pathway leading to IMP production is active during ischemia and adenosine is not the main compound derived from ATP break-down in the human heart. The possible role of extracorporeal circulation is also discussed.

Aged↗

Glucose or essential amino acid infusions in late pregnant and early lactating Simmenthal cows failed to induce a leptin response.

To assess the leptin response to metabolic challenges, three Italian Simmental cows were infused for 6 h: with (a) saline (control); (b) glucose; and (c) amino acid solutions according to a 3 x 3 Latin square experiment. The infusions were carried out at the 36th week of pregnancy, and the second and 12th week of the following lactation. At each of the three infusion periods, blood samples were collected from the jugular vein before and 15, 30, 120, 180, 240, 300 and 360 min after the beginning of each infusion. All samples were analysed for leptin, insulin, glucagon, growth hormone (GH), glucose, non-esterified fatty acids (NEFA) and urea. The physiological phase of the cows significantly affected the basal concentrations of insulin, glucagon, urea and NEFA. The infusion of both glucose and the amino acid solutions did not affect leptin concentrations. Insulin response was significantly increased when animals were infused with the glucose solution and, within treatment, the greatest response was observed at the 12th week of lactation. The greatest glucagon response was observed when infusing the amino acid solution. Urea response to all treatments increased from the dry period to the 12th week of lactation. The GH and NEFA responses were not affected by treatments. The Multi Species radio-immunoassay used in this study showed a lower sensitivity for ruminant leptin which may partially explain the lack of significant leptin variations. However, it can be hypothesized that leptin variations around parturition can be affected by the negative energy balance, and leptin release is not acutely affected by glucose and amino acid availability. In addition, no short-term relationship were found between insulin, glucagon and GH and leptin release in Italian Simmental cows during the dry period and early lactation.

Amino Acids, Essential↗

Influence of dietary conjugated linoleic acid on growth, meat quality, lipogenesis, plasma leptin and physiological variables of lipid metabolism in rabbits.

We investigated the effects of conjugated linoleic acid (CLA) supplementation on growth, feed efficiency, carcass characteristics, meat quality, lipogenesis, and lipid metabolism in rabbits. One hundred forty-four New Zealand White rabbits, half males, half females, age 55 d, mean 1.8 kg BW, were randomly assigned to three weight- and sex-balanced feeding groups in which conventional pelleted diets were supplemented with 0, 0.25, or 0.5% of a CLA preparation. The CLA preparation contained 65% CLA isomers. Twelve rabbits (six males and six females from each group) were slaughtered at each of three slaughtering trials (2.5, 2.8, and 3.1 kg BW, or 76, 90, and 104 d of age). Conjugated linoleic acid supplementation did not influence growth performance (P > or = 0.05) or carcass characteristics but reduced perirenal fat at heavier slaughtering weights (P = 0.09 at 2.8 kg BW; P < 0.01 at 3.1 kg BW). Conjugated linoleic acid reduced acetyl-CoA-carboxylase (CBX) activity in liver (P < 0.05) and adipose tissues (P < 0.01) but did not influence malic enzyme (ME) or glucose-6-phosphate dehydrogenase activity. Significant differences were found between sex in interscapular fat (P < 0.05) for CBX, in perirenal (P < 0.01) and interscapular (P < 0.05) fat for ME, and a tendency (P = 0.070) in liver for glucose-6-phosphate dehydrogenase. The oxidative stability of longissimus lumborum muscle was increased at the higher level of supplementation (P < 0.05). Conjugated linoleic acid reduced (P < 0.05) triglycerides and total cholesterol in plasma with a trend to increased serum leptin (P = 0.06). Plasma triglycerides were higher in males than females (P < 0.01) and plasma leptin tended to be higher in females (2.57 vs. 2.13 ng/ml, P = 0.06). It is concluded that dietary CLA reduced carcass fat in rabbits slaughtered at 2.8 kg or above and altered lipid metabolism to produce lower concentrations of serum triglycerides and total cholesterol and higher concentrations of leptin.

Animal Feed↗

Determination, activity and biological role of adenylosuccinate lyase in blood cells.

Adenylosuccinate lyase deficiency, which is associated with severe mental retardation and autistic features, was discovered in 1984. Since then this enzyme has been analyzed in many human tissues and it is now generally agreed that screening for this enzyme defect should be performed in all unexplained neurological diseases. The aim of the present study was to analyze adenylosuccinate lyase activity in blood cells by a fast simple method adaptable to screening purposes. The activity was also analyzed in B-lymphocytes from patients with B-cell chronic lymphocytic leukemia. The biological role of adenylosuccinate lyase and its importance in regulating cellular levels of AMP is discussed.

Adenine Nucleotides↗

Capillary electrophoresis in the evaluation of ischemic injury: simultaneous determination of purine compounds and glutathione.

An understanding of tissue energy metabolism and antioxidant status is of major interest in the field of organ preservation for transplantation. Nucleotide and glutathione are indicators of cell damage occurring during ischemia and reperfusion. A high performance capillary electrophoresis (HPCE) method with UV detection (185 nm) for the simultaneous analysis of intracellular free ribonucleotides, nucleosides, bases and glutathione (oxidized and reduced form) in myocardial tissues is described. The method does not involve thiol derivatization. The separations were carried out in an uncoated fused-silica capillary, 60 cm long, 52.5 cm to detector, 75 microm ID, with 20 mM Na-borate buffer, pH 10.00, at 20 kV voltage and reading at 185 nm. Injection was hydrostatic for 12 s and total analysis time was 20 min. The technique enables optimum separation of all the compounds examined and has a resolution similar to that of HPLC analysis, with the advantage of fast simultaneous measurement of cell nucleotide metabolism and redox state, not possible with HPLC.

Adolescent↗

Fatty acid composition of phospholipids, triglycerides and cholesterol in serum of castrated and estradiol treated rats.

We have studied the levels of phospholipids, triglycerides, cholesterol esters, and their fatty acid composition in serum for normal, castrated and estradiol treated rats. The sex hormones did not greatly affect the levels of the various lipid fractions which did not undergo great significant variations, under the different treatments. More evident variations occurred in the percent composition of fatty acid and in the content of the various saturated (SAT), unsaturated (UNSAT), essential (EFA) and non essential fatty acids (NEFA). We studied the most important ratios: EFA/NEFA; UNS/SAT; 16:0/16:1; 18:0/18:1, 18:2/18:3; 18:2/20:4. 16:0/16:1; 18:0/18:1 represent the delta9 desaturase, one specific for palmitic, the other for stearic acid. 18:2/18:3 ratio is an index of the delta6 desaturase activity: 18:2/20:4 ratio of delta5 desaturase-elongase. Most changes were evident in triglycerides. We observed a different behaviour of the UNS/SAT and EFA/NEFA ratios in phospholipids and cholesterol esters, which may reflect either an effect of the sex hormones on the exchange of fatty acids between the same lipid fractions, or a redistribution of lipids among different tissues. Great variations were observed of the ratios 16:0/16:1; 18:0/18:1; 18:2/18:3; 18:2/20:4, which are ascribed a different effect of the sex hormones of delta9, delta6, delta5 desaturases.

Animals↗

Restoration of rat liver L-threonine dehydratase activity by pyridoxamine 5'-phosphate: the half-transaminating activity of L-threonine dehydratase and its regulatory role.

When a highly purified preparation of rat liver l-threonine deaminase (l-TDH, EC 4.2.1.16) was 99% inactivated by dialysis, removing bound pyridoxal 5'-phosphate (PLP), the apoenzyme was reactivated not only by PLP but also by pyridoxamine 5'-phosphate (PMP). When purified by HPLC, the commercial PMP used in the incubation mixture was found to contain only extremely small amounts of PLP, which could not account for restoration of l-threonine dehydratase activity. HPLC analysis of the assay mixtures showed that during incubation, sufficient PLP had been formed for reactivation of the apoenzyme. The apoenzyme evidently bound PMP and triggered transamination between PMP and the keto acids, which either contaminated, or were formed by the minimal amount of PLP-holoenzyme always present even in the dialyzed preparation. When sufficient PLP was formed, the PLP-holoenzyme and the original 'true' l-threonine dehydratase activity were restored. When PMP was incubated with the apoenzyme in the presence of small quantities of keto acids (pyruvate or 2-oxobutyrate) small amounts of l-alanine or l-aminobutyrate were formed. The reaction was not reversible; l-alanine and l-aminobutyrate did not react with the PLP-holoenzyme. No transaminating activity occurred with other amino acids. These results show that l-threonine dehydratase exists in two forms: the well known stable apoenzyme-PLP (hydrolase deaminating) and the transient apoenzyme-PMP (non-reversible half-transaminating). Half-transamination has the biological role of keeping the activity of the 'true' l-TDH constant and of regulating intracellular levels of pyruvate, alanine, oxobutyric acid, l-aminobutyric acid, l-threonine and l-serine.

Acetoacetates↗