Oxidative stress in normal-weight obese syndrome.

The normal-weight obese (NWO) syndrome was identified in women whose body weight (BW) and BMI are normal but whose fat mass (FM) is >30%. In these subjects, an early inflammatory status has been demonstrated. The aim was to verify whether oxidative stress occurs in NWO. Sixty age-matched white Italian women were studied and subdivided as follows: 20 normal-weight individuals (NW) (BMI <25 kg/m(2); FM% <30%); 20 NWO (BMI <25 kg/m(2); FM% >30%); 20 preobese-obese (OB) (BMI >25 kg/m(2); FM% >30%). Anthropometric, body composition (by dual-energy X-ray absorptiometry) variables, plasma levels of some cytokines, reduced glutathione (GSH), lipid hydroperoxide (LOOH), nitric oxide (NO) metabolites (NO(2)(-)/NO(3)(-)), antioxidant nonproteic capacity (ANPC) were measured and compared between groups. Glucose and lipid metabolism parameters were assessed. GSH and NO(2)(-)/NO(3)(-) levels resulted lower in OB and NWO compared to NW (P < 0.01). LOOH levels resulted higher in OB and NWO (P < 0.01). ANPC in NWO was lower than NW but higher with respect to OB (P < 0.01). Correlation analysis revealed strong associations between GSH levels and BW, BMI, FM% (R = -0.45, at least P < 0.05); waist circumference (W) (R = -0.33, P < 0.05); FFM% (R = 0.45, P < 0.01); IL-1α, IL-6, IL-10, IL-15 (R = -0.39, -0.33, -0.36 -0.34, respectively, P < 0.05); triglycerides (R = -0.416, P < 0.05). LOOH levels were negatively related to FFM% (R = -0.413, P < 0.05) and positively to FM%, IL-15, TNF-α, insulin, total cholesterol, low-density lipoprotein cholesterol, and triglycerides (R = 0.408, R = 0.502, R = 0.341, R = 0.412, R = 0.4036, R = 0.405, R = 0.405, respectively, P < 0.05). The study clearly indicates that NWO, besides being in early inflammatory status, are contextually exposed to an oxidative stress related to metabolic abnormalities occurring in obesity.

S100B Protein concentration in milk-formulas for preterm and term infants. Correlation with industrial preparation procedures.

Human milk S100B protein possesses important neurotrophic properties. However, in some conditions human milk is substituted by milk formulas. The aims of the present study were: to assess S100B concentrations in milk formulas, to verify any differences in S100B levels between preterm and term infant formulas and to evaluate the impact of industrial preparation at predetermined phases on S100B content. Two different set of samples were tested: (i) commercial preterm (n = 36) and term (n = 36) infant milk formulas; ii) milk preterm (n = 10) and term infant (n = 10) formulas sampled at the following predetermined industrial preparation time points: skimmed cow milk (Time 0); after protein sources supplementation (Time 1); after pasteurization (Time 2); after spray-drying (Time 3). Our results showed that S100B concentration in preterm formulas were higher than in term ones (p < 0.01). In addition, S100B concentrations during industrial preparation showed a significant increase (p < 0.001) at Time 1 followed by a slight decrease (p > 0.05) at Time 2, whereas a significant (p < 0.001) dip was observed at Time 3. In conclusion, S100B showed a sufficient thermostability to resist pasteurization but not spry-drying. New feeding strategies in preterm and term infants are therefore warranted in order to preserve S100B protein during industrial preparation.

Maternal dietary habits and mycotoxin occurrence in human mature milk.

During 2006, 82 samples of human mature milk were collected at Italian hospitals and checked for aflatoxin M1 (AFM1) and ochratoxin A (OTA) by immunoaffinity column extraction and HPLC. AFM1 was detected in four (5%) of milk samples (ranging from < 7 ng/L to 140 ng/L; mean level: 55.35 ng/L); OTA was detected in 61 (74%) of milk samples (ranging from < 5 ng/L to 405 ng/L; mean level: 30.43 ng/L. OTA levels were significantly higher (p less, not double equals 0.05) in milk of habitual consumers of bread, bakery products and cured pork meat. No other statistically significant differences were observed although habitual consumers of pasta (p = 0.059), cookies (p = 0.061) and juices (p = 0.063) had mean contamination values of OTA higher than the moderate consumer. The very few AFB1 positive samples did not allow statistical comparisons. The present study confirms that the occurrence of OTA in human milk is related to maternal dietary habits. The findings support the possibility of dietary recommendations to woman, during pregnancy and lactation, aimed to tentatively reduce the OTA contamination of human milk.

Response of cell cycle/stress-related protein expression and DNA damage upon treatment of CaCo2 cells with anthocyanins.

Anthocyanins are a class of flavonoids, widely spread throughout the plant kingdom, exhibiting important antioxidant and anti-inflammatory actions as well as chemotherapeutic effects; nonetheless, little is known about the molecular mechanisms by which these activities are exerted. The present study is aimed at investigating molecular mechanisms involved in the chemotherapeutic effects induced by both cyanidin-3-O-beta glucopyranoside (CY3G) and its aglycon form, cyanidin chloride (CY), in human colon cancer cells (CaCo2). The effect on cell growth, reactive oxygen species (ROS) formation and cell cycle/stress proteins modification, including ataxia teleangectasia mutated protein (ATM), p53, p21, 8-oxoguanine DNA glycosylase (OGG1), 70 kDa heat shock protein (HSP70) and topoisomerase IIbeta, as well as on DNA fragmentation, was determined. CY and CY3G treatment affect cell growth and cell proliferation, this latter in a moderately dose-dependent way. Interestingly, ROS level is decreased by any concentration of CY and, only at the lowest concentration, by CY3G. Moreover, the two molecules exert their activities increasing ATM, topoisomerase II, HSP70 and p53 expression. The analysis of DNA fragmentation by Comet assay evidences: (1) a dose-dependent increase in DNA damage only after treatment with CY3G; (2) a more evident trend in the DNA fragmentation when the treatment is performed on agarose embedded cells (cellular atypical Comet); (3) a highly dose-dependent DNA fragmentation induced by CY when the treatment is carried out on agarose embedded naked DNA (acellular atypical Comet). The present findings substantiate a possible chemotherapeutic role of anthocyanins and suggest that CY and CY3G act on CaCo2 by different mechanisms, respectively, ROS-dependent and ROS-independent.

Phenolic compounds and antioxidant activity of Italian extra virgin olive oil Monti Iblei.

The profile of phenolic compounds, antioxidant capacity, oxidative stability, and chemical characteristics (free acidity, peroxide value, specific extinction K232 and K270 values, and DeltaK) of 22 commercial extra virgin olive oil (EVOO) samples coming from the denomination of protected origin (DPO) Monti Iblei and obtained from olives harvested in the period September-December 2005 in the production area of the province of Siracusa (Sicily, Italy) were evaluated. The content of total phenols, expressed as gallic acid equivalents, ranged from 14.80 to 121.20 mg/100 g, with a mean value of 53.72 mg/100 g, mainly attributable to deacetoxyligstroside aglycone, deacetoxyoleuropein aglycone, oleuropein aglycone, and ligstroside aglycone. The mean values of Trolox equivalent antioxidant capacity (TEAC) and of oxidative stability were 54.76 and 11.99 hours, respectively. Both TEAC and oxidative stability were positively correlated to the phenol content and to the percentage of inclusion of the olive cultivar "Tonda Iblea." The high mean content of phenols, besides conferring prolonged oxidative stability, likely confers to the DPO Monti Iblei EVOO marked potential beneficial effects for human health.

Protective effect of cyanidin 3-O-beta-D-glucoside on ochratoxin A-mediated damage in the rat.

The aim of the present study was to verify whether the oral administration of cyanidin 3-O-beta-D-glucoside (C3G) might counteract damage induced by chronic exposure (28 d) to ochratoxin A (OTA) in rats and if its effect may be mediated by haeme oxygenase-1 (HO-1). Forty male Sprague-Dawley rats, individually caged, were divided into four groups of ten animals. A control group received a commercial diet, group C3G received the control diet supplemented with C3G (1 g/kg feed), group OTA received the control diet supplemented with 200 parts per billion of OTA, and group OTA+C3G received the OTA group diet supplemented with C3G (1 g/kg feed). After 4 weeks of treatment animals were killed and the liver, kidneys and brain of each rat were collected and homogenised to evaluate non-proteic thiol groups (RSH), lipid hydroperoxide (LOOH) levels, HO-1 expression and DNA fragmentation. Rats of the OTA group showed a significant (P < 0.001) decrease in RSH content of kidney and liver and a significant (P < 0.001) increase of LOOH in all the examined tissues compared with the control group. In the OTA+C3G group both RSH content and LOOH levels were similar to those observed in the control group, demonstrating that C3G was able to counteract the effects of OTA. A significant (P < 0.001) induction of HO-1 was evident in kidney and liver of both OTA and C3G groups. DNA damage occurred in all the examined tissues of the OTA group, whereas C3G was able to prevent it. The present study confirmed that the effects of OTA are mediated by oxidative stress and demonstrated that C3G efficiently counteracted deleterious effects of OTA because of its antioxidant and HO-1-inducing properties.

Bioavailability, antioxidant and biological properties of the natural free-radical scavengers cyanidin and related glycosides.

Cyanidin and its glycosides (Cy and Cyg) have been indicated as promising candidates as dietary compounds with a potential role in human health. They are the largest class of water-soluble compounds in plants, where they are responsible for the brilliant color (red, orange, blue) of fruits and flowers. As natural compounds of several foods such as vegetables, fruits and red wines, they are estimated to be widely ingested by humans. This paper, basing on the data previously reviewed in 2002, focuses on the findings regarding human and animal studies on Cy and Cyg absorption and metabolism, antioxidant activity and biological properties, with particular attention to anticarcinogenic activity, vasoprotective, anti-inflammatory, anti-obesity and anti-diabetes effects. It is concluded that although Cy and Cyg bioavailability is low, further investigations are necessary because some important metabolites may still not have been identified. Literature data on antioxidant activity and biological properties, however, widely confirm Cy and Cyg as dietary compounds with a potential beneficial role in human health.

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.

Survey of the occurrence of Aflatoxin M1 in ovine milk by HPLC and its confirmation by MS.

During the period of October-July 2000, 240 samples of dairy ewes milk, obtained from farms of Enna (Sicily, Italy), were checked for Aflatoxin M(1) (AFM(1)) by HPLC using a fluorimetric detector. The limit of detection and the limit of quantification were 250 ng/L for AFM(1). All the positive milk samples for AFM(1) were confirmed by LC-MS. AFM(1) was detected in 81% of milk samples, ranging from 2 to 108 ng/L. Three samples were over the legal limits (50 ng/L). Mean contamination of samples obtained from stabulated ewes was higher than that from grazing ewes (35.27 vs. 12.47 ng/L). Furthermore, samples collected in the period September-October showed higher contamination than samples collected during the other months (42.68 vs. 10.55 ng/L). Both differences are related to the administration of compound feed. Based on current toxicological knowledge we concluded that the AFM(1) contamination levels recorded in ewe milk did not present a serious human health hazard. However, as ewe milk is exclusively used to produce cheese due to its higher protein content, and also considering the preferential binding of AFM(1) to casein during coagulation of milk, a potentially high concentration effect could occur, thus the surveillance of contamination levels should be more continuous and widespread.

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.

Ochratoxin A-induced DNA damage in human fibroblast: protective effect of cyanidin 3-O-beta-d-glucoside.

Ochratoxin A (OTA), a mycotoxin produced by Aspergillus ochraceus and other moulds, has recently received growing attention because of its carcinogenic, teratogenic and nephrotoxic properties in both humans and farm animals. Nevertheless, with regard to the mechanism of toxicity, the data in the literature are inconclusive. The aim of our work was to verify in human fibroblasts treated with different OTA dosages the involvement of oxidative pathway in the damage mechanism of this mycotoxin and the possible protective effect exerted by cyanidin 3-O-beta-D-glucoside (C3G), an anthocyanin present in pigmented oranges, red wines, fruits and vegetables. The addition of OTA at 25 and 50 microM concentrations for 48 h determined only a slight but significant (P<.05) increase in radical oxygen species, whereas a substantial increase in their production was observed at longer exposure, in particular, when the fibroblasts were treated with 50 microM OTA for 72 h. Under the same experimental conditions, our data showed a significant (P<.05) increase in the rupture of cellular membrane and high damage to genomic DNA, evaluated by single-cell gel electrophoresis (comet assay), thus confirming the involvement of oxidative stress in the OTA genotoxicity in agreement with other studies. Diversely, mitochondrial functionality does not appear influenced by OTA treatment. C3G (0.125, 0.250 mM) added to the cells treated with 50 microM OTA significantly reduced free radical species production and prevented genomic DNA damage.

Cyanidins: metabolism and biological properties.

Cyanidin and its glycosides belong to the anthocyanins, a widespread class of water-soluble plant compounds that are responsible for the brilliant color (red, orange, blue) of fruits and flowers. They are widely ingested by humans as it has been estimated a daily intake around 180 mg, mainly deriving from fruits and red wines. This paper reviews the literature on the biological activities, absorption and metabolism of cyanidins, with emphasis to the antioxidant, antimutagenic and other protective activities ascribed to these compounds. Their role in contrasting development of cancer and other pathologies is also reviewed. It is concluded that a great deal of work is still necessary to i) definitively clarify the metabolism of cyanidins in human beings; ii) assess the dietary burden and variations within and between populations; iii) evaluate the relationship between cyanidin glycosides-rich food consumption and incidence of given pathologies. The amount of work to be performed is even more significant when considering a possible therapeutic use of cyanidin glycosides-based drugs. With this aim, information on absorption, distribution, metabolism and excretion of cyanidin-glycosides administered by main possible routes are largely insufficient. However, consisting findings allow looking at cyanidins as dietary compounds with a potential beneficial role for human health.