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Biomedical subjects

Vincenzo Fogliano

Publications and source records attributed to Vincenzo Fogliano.

At least 19 recordsLinked to original sources

Treatment of cereal products with a tailored preparation of trichoderma enzymes increases the amount of soluble dietary fiber.

Nutritionists recommend increasing the intake of soluble dietary fiber (SDF), which is very low in most cereal-based products. Conversion of insoluble DF (IDF) into SDF can be achieved by chemical treatments, but this affects the sensorial properties of the products. In this study, the possibility of getting a substantial increase of SDF from cereal products using a tailored preparation of Trichoderma enzymes is reported. Enzymes were produced cultivating Trichoderma using durum wheat fiber (DWF) and barley spent grain (BSG) as unique carbon sources. Many Trichoderma strains were screened, and the hydrolysis conditions able to increase by enzymatic treatment the amount of SDF in DWF and BSG were determined. Results demonstrate in both products that it is possible to triple the amount of SDF without a marked decrease of total DF. The enzymatic treatment also causes the release of hydroxycinnamic acids, mainly ferulic acid, that are linked to the polysaccharides chains. This increases the free phenolic concentration, the water-soluble antioxidant activity, and, in turn, the phenol compounds bioavailability.

Coumaric Acids↗

Study of the three-way interaction between Trichoderma atroviride, plant and fungal pathogens by using a proteomic approach.

The main molecular factors involved in the complex interactions occurring between plants (bean), two different fungal pathogens (Botrytis cinerea, Rhizoctonia solani) and an antagonistic strain of the genus Trichoderma were investigated. Two-dimensional (2-D) electrophoresis was used to analyze separately collected proteomes from each single, two- or three-partner interaction (i.e., plant, pathogenic and antagonistic fungus alone and in all possible combinations). Differential proteins were subjected to mass spectrometry and in silico analysis to search for homologies with known proteins. In the plant proteome, specific pathogenesis-related proteins and other disease-related factors (i.e., potential resistance genes) seem to be associated with the interaction with either one of the two pathogens and/or T. atroviride. This finding is in agreement with the demonstrated ability of Trichoderma spp. to induce systemic resistance against various microbial pathogens. On the other side, many differential proteins obtained from the T. atroviride interaction proteome showed interesting homologies with a fungal hydrophobin, ABC transporters, etc. Virulence factors, like cyclophilins, were up-regulated in the pathogen proteome during the interaction with the plant alone or with the antagonist too. We isolated and confidently identified a large number of protein factors associated to the multi-player interactions examined.

Amino Acid Sequence↗

Glycated fibroblast growth factor-2 is quickly produced in vitro upon low-millimolar glucose treatment and detected in vivo in diabetic mice.

Angiogenesis impairment in hyperglycemic patients represents a leading cause of severe vascular complications of both type-1 and -2 diabetes mellitus (DM). Angiogenesis dysfunction in DM is related to glycemic control; however, molecular mechanisms involved are still unclear. Fibroblast growth factor-2 (FGF-2) is a potent angiogenic factor and, according to previous evidence, may represent a key target of molecular modifications triggered by high-sugar exposure. Therefore, the purpose of this study was to investigate whether short incubation with hyperglycemic levels of glucose affected FGF-2 and whether glucose-modified FGF-2 was detectable in vivo. Biochemical analyses carried out with SDS-PAGE, fluorescence emission, mass-spectrometry, immunoblot, and competitive ELISA experiments demonstrated that human FGF-2 undergoes a rapid and specific glycation upon 12.5-50 mm glucose exposure. In addition, FGF-2 exposed for 30 min to 12.5 mm glucose lost mitogenic and chemotactic activity in a time- and dose-dependent manner. Under similar conditions, binding affinity to FGF receptor 1 was dramatically reduced by 20-fold, as well as FGF receptor 1 and ERK-1/2 phosphorylation, and FGF-2 lost about 45% of angiogenic activity in two different in vivo angiogenic (Matrigel and chorioallantoic-membrane) assays. Such glucose-induced modification was specific, because other angiogenic growth factors, namely platelet-derived growth factor BB and placental-derived growth factor were not significantly or markedly less modified. Finally, for the first time, glycated-FGF-2 was detected in vivo, in tissues from hyperglycemic nonobese diabetic mice, in significantly higher amounts than in normoglycemic mice. In conclusion, hyperglycemic levels of glucose may strongly affect FGF-2 structure and impair its angiogenic features, and endogenous glycated-FGF-2 is present in diabetic mice, indicating a novel pathogenetic mechanism underlying angiogenesis defects in DM.

Animals↗

Effect of cooking on the concentration of Vitamins B in fortified meat products.

B vitamins fortification of meat products is useful to compensate the loss of these compounds occurring during the heat treatment. The objective of this study was to evaluate the influence of heat treatments on the B vitamins concentration in fortified meat products. A rapid and reliable method for the simultaneous determination of Vitamins B(1), B(6) and B(12) in homogenized boiled ham and in various fortified burgers was set up. Extraction procedure and HPLC method ensure low detection limits, good sensitivity and resolution. Results showed that cooking processes caused a decrease in the B vitamins content both in mild (70-90 degrees C) and severe (120 degrees C) conditions. Performing a fortification of 25 microg g(-1) the residual concentration of B vitamins after cooking allow to reach the recommended daily allowance, thus suggesting that B vitamins fortification of meat product is an useful practice.

Animals↗

Glycation of lysine-containing dipeptides.

Protein glycation through Maillard reaction (MR) is a fundamental reaction both in foods and in the human body. The first step of the reaction is the formation of Amadori product (AP) that is converted into intermediate and advanced MR products during reaction development. Although the MR is not an enzymatic reaction, a certain degree of specificity in the glycation site has been observed. In the present study, we have monitored the glycation of different lysine-containing dipeptides to evaluate the influence on the NH(2) reactivity of the neighboring amino acid.Lysine dipeptides were reacted with glucose, galactose, lactose and maltose. The formation and identification of glycated compounds were monitored by mass spectrometry (MALDI-TOF and ESI-MS/MS) and by HPLC of their Fmoc derivatives. MS/MS analysis showed that the glucose APs formed on dipeptides have a characteristic fragmentation pattern: the fragment at [M - 84](+) due to the formation of pyrylium and furylium ion is mainly present in the monoglucosylated form, while the [M - 162](+) and the [M - 324](+) are more evident in the fragmentation pattern of the diglucosylated forms. The nature of the vicinal amino acids strongly affects lysine reactivity towards the different carbohydrates: the presence of hydrophobic residues such as Ile, Leu, Phe strongly increases lysine reactivity. Contrasting results were obtained with basic residues. The Lys-Arg dipeptide was among the most reactive while the Lys-Lys was not.

Dipeptides↗

Absorption and metabolism of red orange juice anthocyanins in rats.

Anthocyanins are natural pigments that could be involved in various health effects. Red oranges are an important dietary source of anthocyanins, including cyanidin 3-glucoside (Cy 3-glc) and an acylated derivative, cyanidin 3-(6''-malonyl)-glucoside (Cy 3-malglc). The aim of this study was to evaluate the absorption and metabolism of red orange anthocyanins in rats fed an anthocyanin-enriched diet for 12 d (approximately 2.8 micromol anthocyanins/d). Furthermore, the absorption of these anthocyanins was studied in both the stomach and intestine using in situ models in rats. Anthocyanin metabolites were identified and quantified by HPLC-electrospray ionization tandem MS and HPLC-diode array detection, respectively. The red orange anthocyanins, Cy 3-glc and Cy 3-malglc, as well as their respective methylated derivatives, were recovered in urine after red orange juice intake. The 24 h urinary excretion of total anthocyanins was low (0.081 (SEM 0.009) % of the ingested amount). However, a high proportion (about 20 %) of red orange anthocyanins was absorbed from the stomach. More Cy 3-malglc than Cy 3-glc was absorbed in the intestine. This study thus indicated that red orange juice anthocyanins were rapidly absorbed from both stomach and small intestine, and then excreted in the urine as intact and methylated forms. Moreover, the absorption and metabolism of acylated anthocyanins and non-acylated anthocyanins were similar.

Acylation↗

Apples increase nitric oxide production by human saliva at the acidic pH of the stomach: a new biological function for polyphenols with a catechol group?

Dietary inorganic nitrate is secreted in saliva and reduced to nitrite by bacterial flora. At the acidic pH of the stomach nitrite is present as nitrous acid in equilibrium with nitric oxide (*NO), and other nitrogen oxides with nitrating and nitrosating activity. *NO in the stomach exerts several beneficial effects, but nitrosating/nitrating species have been implicated as a possible cause of epithelial neoplasia at the gastroesophageal junction. We investigated the effects of apple extracts on *NO release by human saliva at pH 2. A water extract obtained from apple homogenate increased *NO release caused by acidification of saliva. Data show that polyphenols were responsible for this activity, with chlorogenic acid and (+)-catechin the most active and concentrated species. However, ferulic acid, a hydroxycinnamic acid with only one aromatic hydroxyl group, did not increase *NO release. Fructose, the most representative sugar in apples, was also inactive. Interestingly, ascorbic acid in saliva induced a SCN(-)-enhanced burst of *NO but, unlike apple, the release was transient. The simultaneous addition of ascorbic acid and apple extract caused a burst of *NO followed by the increased steady-state level characteristic of saliva containing apple extract. Chlorogenic acid and (+)-catechin, but not ferulic acid, formed o-semiquinone radicals and nitrated polyphenols, suggesting the scavenging of *NO(2) by o-semiquinones. Our results propose that some apple polyphenols not only inhibit nitrosation/nitration but also promote *NO bio-availabilty at the gastric level, a previously unappreciated function.

Antioxidants↗

Thermal degradation studies of food melanoidins.

Food melanoidins were isolated from bread crust, coffee, and tomato sauce and their composition was investigated by thermal degradation. Among the generated volatiles, important food flavor compounds were detected: in particular furans, carbonyl compounds, 1,3-dioxolanes, pyrroles, pyrazines, pyridines, thiophenes, and phenols. The results indicated that the isolated melanoidin fractions mainly consisted of compounds formed from carbohydrates and their degradation products. Besides proteins, other food constituents were incorporated in the melanoidin structure as well, such as lipid oxidation products in tomato melanoidins and phenolic compounds in coffee melanoidins. A comparison of the thermal generation of volatiles between these food-derived melanoidins and model melanoidins prepared from a single carbonyl compound and an amino acid showed that the degradation pattern of food melanoidins is quite different from that obtained from a glucose-glycine model system.

Bread↗

Optothermistor as a breakthrough in the quantification of lycopene content of thermally processed tomato-based foods: verification versus absorption spectrophotometry and high-performance liquid chromatography.

This study reports on the first use of the "optothermistor" as a novel, precise, fast, and low-cost detector of lycopene in a wide range of commercially available processed-tomato products. The quantitative performance of the new device was evaluated by comparing data obtained to that acquired by conventional methods, namely, absorption spectrophotometry and high-performance liquid chromatography (HPLC); the linear correlation was high (R = 0.98). The variation of data obtained with the optothermistor in a series of consecutive measurements performed with the same loading of the sample was better than 1%. However, the repeatability (RSD 0.5-9.0%, n = 3-5) achieved with the optothermistor by independent analyses (multiple loading) is comparable to that of HPLC and spectrophotometry. Results of the studies performed on the 19 products derived from tomatoes demonstrated that the optothermistor is suitable for selective, accurate, precise, and simple determination of lycopene (range = 7-75 mg/100 g of product weight) without the need for a sample pretreatment step. The estimated sensitivity of the present optothermistor is 2 mg of lycopene/100 g of product.

Carotenoids↗

Changes in carotenoid and ascorbic acid contents in fruits of different tomato genotypes related to the depletion of UV-B radiation.

The aim of the present study was to investigate if the depletion of UV-B radiation affected the most representative carotenoids as well as the ascorbic acid content in tomato fruits, harvested at both breaker and firm red stages. To do this, three tomato genotypes, DRW 5981, HP 1, and Esperanza, were grown inside a greenhouse either covered with polyethylene transparent to UV-B or depleted of UV-B by a special covering film. The antioxidant properties of the fruits were evaluated on the water-insoluble fractions according to the ABTS method. UV-B effect on antioxidant activity was negligible in DRW and HP 1 genotypes, whereas it was detrimental in Esperanza at both ripening stages. This genotype seems to have a negligible capability of accumulating carotenoids and a great susceptibility to detrimental effects of UV-B; conversely, the DRW genotype shows high carotenoid levels under sunlight conditions and a further promotion by UV-B. On the other hand, the HP 1 mutant displays an intermediate behavior and represents the only genotype favored by UV-B with respect to ascorbic acid accumulation.

Antioxidants↗

Functional foods: planning and development.

The discussion on functional foods among scientific communities and EC commissions is mainly focused on definitions and on legislative issues related to the presence on the market of this kind of products. Although many new products continuously appear, the functional food market is characterized by a high rate of failure. To realize a successful functional food, experts having different background should work together following a detailed workplan. In this paper, the problems related to planning and development of functional foods are considered using a step-by-step approach. The strategies for invention and development, formulation and validation of nutritional claims are regarded also illustrating practical examples of functional food development. The concept of food for special medical purposes, intended for individuals who are being treated under medical supervision, is also introduced.

Chemical Phenomena↗

Bioavailability of trans-resveratrol from red wine in humans.

Many in vitro studies demonstrated significant biological effects of trans-resveratrol. Thus, understanding the rate of intestinal absorption and metabolization in vivo of trans-resveratrol is the prerequisite to evaluate its potential health impact. Bioavailability studies mainly in animals or in humans using the pure compound at very high doses were performed. In this work, trans-resveratrol bioavailability from a moderate consumption of red wine in 25 healthy humans has been studied by three different experiments. The wine ingestion was associated to three different dietary approaches: fasting, a standard meal, a meal with high and low amount of lipids. Trans-resveratrol 3- and 4'-glucuronides were synthesized, purified, and characterized as pure standards. Bioavailability data were obtained by measuring the concentration of free, 3-glucuronide and 4'-glucuronide trans-resveratrol by high-performance liquid chromatography (HPLC), both with ultraviolet (UV) and mass spectrometry (MS) detection, in serum samples taken at different times after red wine administration. Free trans-resveratrol was found, in trace amounts, only in some serum samples collected 30 min after red wine ingestion while after longer times resveratrol glucuronides predominated. Trans-resveratrol bioavailability was shown to be independent from the meal or its lipid content. The finding in human serum of trans-resveratrol glucuronides, rather than the free form of the compound, with a high interindividual variability, raises some doubts about the health effects of dietary resveratrol consumption and suggests that the benefits associated to red wine consumption could be probably due to the whole antioxidant pool present in red wine.

Adult↗

Bread crust melanoidins as potential prebiotic ingredients.

Melanoidins are the final products of the Maillard reaction. They are a heterogeneous mixture of compounds characterized by brown color and high molecular weight. The physiological properties of melanoidins have been widely investigated and there is a general consensus on their poor digestibility and bioavailability. In vitro studies on food melanoidins are in many cases limited by their poor water solubility. This problem was recently overcome for bread melanoidins using an enzymatic digestion procedure. Bread melanoidins are constituted by low-molecular-weight, colored compounds linked to the gluten polymer. In this work, melanoidins from different bread types were investigated for their potential prebiotic activity by a static batch culture. Results showed that anaerobic bacteria, particularly Bifidobacteria strains, are able to use bread melanoidins as carbon source. The bacterial growth is different for the various types of melanoidins samples indicating that starting materials and processing conditions have a strong influence on the prebiotic potential of bread melanoidins. In all cases the bacterial growth obtained using bread melanoidins is lower than that previously observed using melanoidins from other sources, such as coffee silverskin.

Bacteroides↗

Antioxidant activity of pasteurized and sterilized commercial red orange juices.

Blood orange juice is a typical Italian product whose red color is primarily associated with anthocyanin pigments. Two orange-based products are present on the market: pasteurized pure juice with 40 days of shelf life, and sterilized beverage containing minimum 12% of concentrated fruit juice. The aim of the present paper is to verify the relationships between the antioxidant properties and the anthocyanins content in a sampling of pasteurized and sterilized commercial red orange juices. The anthocyanins composition was determined by HPLC-MS/MS, while the antioxidant activity was evaluated by the Briggs-Rauscher reaction, selected in order to acquire information at acid pH values, by three radical scavenging assays (DMPD, 2-2'-azinobis-(3-ethylenbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), DPPH), and by FRAP assay to monitor the ferric reducing power. Results showed that antioxidant activity, particularly when measured by ABTS method, is positively related to the content of anthocyanins and that the reduction of anthocyanins content, typical of commercial long-shelf life juices, leads to a remarkable loss of antioxidant power.

Anthocyanins↗

Novel cyclic lipodepsipeptide from Pseudomonas syringae pv. lachrymans strain 508 and syringopeptin antimicrobial activities.

The syringopeptins are a group of antimicrobial cyclic lipodepsipeptides produced by several plant-associated pseudomonads. A novel syringopeptin, SP508, was shown to be produced as two homologs (A and B) by Pseudomonas syringae pv. lachrymans strain 508 from apple and to structurally resemble syringopeptin SP22. SP508 differed from SP22 and other syringopeptins by having three instead of four alpha,beta-unsaturated amino acids and a longer beta-hydroxy acyl chain. Both SP508 and SP22 displayed growth-inhibitory activities against Mycobacterium smegmatis, other gram-positive bacteria, and yeasts but not against gram-negative bacteria. Structure-activity analyses of the SP508 and SP22 homologs indicated chemical structural features that lead to enhanced antimycobacterial activity by these pseudomonad cyclic lipodepsipeptides.

Anti-Infective Agents↗

Substrate specificity of amadoriase I from Aspergillus fumigatus.

Generation of Amadori products is the major single modification by the Maillard reaction in vivo and a source of biologically active glycoxidation products leading to protein cross-linking in biological tissues. Amadoriase I from Aspergillus fumigatus cleaves Amadori products into deoxyglucosone, hydrogen peroxide, and the corresponding primary amine. It has been reported that Amadori products formed on free amino acids are a good substrate for amadoriase I, whereas the enzyme is unable to cleave Amadori products formed on whole proteins. This work aims to investigate the affinity of amadoriase I for oligopeptides and small proteins. Recombinant amadoriase I was expressed in E. coli and purified by Ni His-tag affinity chromatography. Di-, tri-, and tetrapeptides were derivatized with glucose, and the corresponding Amadori products were purified by TLC and HPLC. Glycated beta-lactoglobulin was also used as a substrate. In both cases the formation of Amadori products was monitored by electrospray ionization-mass spectrometry (ESI-MS). The Km of amadoriase for glycated-L-lysine was 4.2 mM, which is in accordance with the literature. Km decreases with the length of peptide, being slightly reduced for dipeptides, and is around 10 mM for tri- and tetrapeptides. Glycated proteins are not substrates of the enzyme; but when amadoriase I was added during the glycation reaction, a significant reduction of Amadori product formation was observed on both peptides and proteins. Data confirm the hypothesis that steric hindrance is critical for amadoriase I activity, indicating that oligopeptides up to four amino acids in length are good substrates. Moreover, these data show that, at least in some experimental conditions, amadoriase I is able to reduce the formation of protein-bound Amadori product.

Amino Acid Oxidoreductases↗

Structure, conformation and biological activity of a novel lipodepsipeptide from Pseudomonas corrugata: cormycin A.

Cationic lipodepsipeptides from Pseudomonas spp. have been characterized for their structural and antimicrobial properties. In the present study, the structure of a novel lipodepsipeptide, cormycin A, produced in culture by the tomato pathogen Pseudomonas corrugata was elucidated by combined protein chemistry, mass spectrometry and two-dimensional NMR procedures. Its peptide moiety corresponds to L-Ser-D-Orn-L-Asn-D-Hse-L-His-L-aThr-Z-Dhb-L-Asp(3-OH)-L-Thr(4-Cl) [where Orn represents ornithine, Hse is homoserine, aThr is allo-threonine, Z-Dhb is 2,3-dehydro-2-aminobutanoic acid, Asp(3-OH) is 3-hydroxyaspartic acid and Thr(4-Cl) is 4-chlorothreonine], with the terminal carboxy group closing a macrocyclic ring with the hydroxy group of the N-terminal serine residue. This is, in turn, N-acylated by 3,4-dihydroxy-esadecanoate. In aqueous solution, cormycin A showed a rather compact structure, being derived from an inward orientation of some amino acid side chains and from the 'hairpin-bent' conformation of the lipid, due to inter-residue interactions involving its terminal part. Cormycin was significantly more active than the other lipodepsipeptides from Pseudomonas spp., as demonstrated by phytotoxicity and antibiosis assays, as well as by red-blood-cell lysis. Differences in biological activity were putatively ascribed to its weak positive net charge at neutral pH. Planar lipid membrane experiments showed step-like current transitions, suggesting that cormycin is able to form pores. This ability was strongly influenced by the phospholipid composition of the membrane and, in particular, by the presence of sterols. All of these findings suggest that cormycin derivatives could find promising applications, either as antifungal compounds for topical use or as post-harvest biocontrol agents.

Amino Acid Sequence↗