Luciana Azevedo

Black bean (Phaseolus vulgaris L.) as a protective agent against DNA damage in mice

This study was designed to evaluate the toxicogenetic or protective effect of cooked and dehydrated black beans (Phaseolus vulgaris L.) in bone marrow and peripheral blood cells of exposed mice. The frequency of micronuclei detected using the bone marrow erythrocyte micronucleus test and level of DNA lesions detected by the comet assay were chosen as end-points reflecting mutagenic and genotoxic damage, respectively. Initially, Swiss male mice were fed with a 20% black bean diet in order to detect mutagenic and genotoxic activity. However, no increase in the frequency of bone marrow micronucleated polychromatic erythrocytes (MN PCEs) or DNA lesion in leukocytes was observed. In contrast, received diets containing 1, 10 or 20% of black beans, a clear, but not dose-dependent reduction in the frequency of MN PCEs were observed in animals simultaneously treated with cyclophosphamide, an indirect acting mutagen. Similar results were observed in leukocytes by the comet assay. Commercial anthocyanin was also tested in an attempt to identify the bean components responsible for this protective effect. However, instead of being protective, the flavonoid, at the highest dose administered (50 mg/kg bw), induced primary DNA lesion, as detected by the comet assay. These data indicate the importance of food components in preventing genetic damage induced by chemical mutagens, and also reinforce the role of toxicogenetic techniques in protecting human health.

Influence of Marolo (Annona crassiflora Mart.) Pulp Intake on the Modulation of Mutagenic/Antimutagenic Processes and Its Action on Oxidative Stress In Vivo

Brazilian savanna constitutes a valuable ecoregion that contributes to the supply of fruit known worldwide for its nutritional value and peculiar flavors. Among them, the Marolo (Annona crassiflora Mart.) fruit is known for its use in folk medicine. In order to establish the safety of Marolo for human consumption, this study evaluated the following: the chemical composition of Marolo pulp; its mutagenic and antimutagenic activities using micronucleus test; and the oxidative stress induced in the livers of mice fed a diet containing 1%, 10% or 20% pulp. It was observed that the chemical composition of marolo pulp was similar to that of common fruit; nevertheless, its lipidic content and energetic values were higher. In the mice fed experimental diets, the biochemical parameters of the blood serum showed normal levels of glucose, triglycerides, and cholesterol. The micronucleus test indicated neither mutagenic nor antimutagenic effects of Marolo consumption on bone marrow cells but showed potentialization of cyclophosphamide (CP). The oxidative stress levels observed indicated that CP was not exerting a great influence on the induction of reactive oxygen species. As the whole fruit is a complex matrix, the interactions between its components could be responsible for its negative and positive biological effects.

Organic and Conventional Coffea arabica L.: A Comparative Study of the Chemical Composition and Physiological, Biochemical and Toxicological Effects in Wistar Rats

Differentiation between organic and conventional coffee has increased due to the growing demand and high consumption of healthy foods that contain compounds with antioxidant potential, which have been associated with the reduction of chronic diseases. We used organic and conventional coffee in powder 4% (w/w) and infusions 5%, 10% and 20% (w/v) incorporated in a commercial diet to test in vivo. The levels of chlorogenic acid, caffeine and trigonelline were determined by high performance liquid chromatography. The body weight, weight gain, food consumption, aberrant foci crypt, mucin depleted foci, stress biomarkers protein carbonyl and malondialdehyde, biochemical parameters and behavior of the rats were compared between the experimental and control groups within a framework of colon carcinogenesis. The organic coffee showed higher levels of chlorogenic acid, caffeine and trigonelline than conventional, however, this difference did not significantly affect behavior. The infusions had an antioxidant effect, reducing the levels of malondialdehyde; however, the biochemical parameters of the serum were not altered, and there was neither induction nor prevention of preneoplasic lesions.

The addition of inulin and Lactobacillus casei 01 in sheep milk ice cream

The effect of the Lactobacillus casei 01 and inulin addition on sheep milk ice cream during storage (-18 °C, 150 days) was investigated. Control, probiotic and synbiotic ice cream (10% w/w sheep milk cream; 10% w/w sheep milk cream, L. casei 01, 6 log CFU/mL; 10% w/w inulin, L. casei 01, 6 log CFU/mL, respectively) were manufactured. Microbiological counts (probiotic count, survival after in vitro gastrointestinal resistance, Caco-2 cell adhesion), bioactivity and microstructure were analysed. Physical and textural characteristics, colour parameters, thermal analysis and organic acids/volatile compounds were also evaluated. All formulations supported L. casei 01 viability and maintained above the minimum therapeutic level (>6 log CFU/mL) during storage. Inulin did not affect L. casei 01 survival after the passage through simulated gastrointestinal tract and adhesion to Caco-2 cells while improved the ACE-inhibitory and antioxidant activity. L. casei 01 addition produced several volatile compounds, such as carboxylic acids, alcohols, aldehydes and ketones. Also, scanning electron microscopy showed an interaction between probiotic bacteria and inulin fibre on synbiotic ice cream and the adhesion of L. casei to Caco-2 cells was observed.

First evaluation of the antimutagenic effect of mangaba fruit in vivo and its phenolic profile identification

The chemical composition and functional effects of mangaba fruit pulp were evaluated through a multi-endpoint assay in mice, consisting of the bone marrow micronucleus test, gut micronucleus test, and the apoptosis, oxidative stress, and comet assays. Mangaba fruit pulp was administered in three doses, 10, 20, and 40ml/kg body weight (b.w.), by gavage to male Swiss mice against doxorubicin and dimethylhydrazine-induced mutagenicity. The phenolic profile of the mangaba fruit pulp was evaluated by HPLC, and seven compounds were identified: gallic acid, catechin, chlorogenic acid, vanillic acid, o-coumaric acid, rosmarinic acid, and rutin. The in vivo tests revealed that mangaba fruit pulp showed no toxic/mutagenic effects in any of the assays performed, and also showed protective effects at all endpoints. At the three administered extract concentrations, the main results about the protective effects were as follows: bone marrow micronucleus test (42.33, 58.14, and 77.21%), micronucleus gut test (34.21, 63.15, and 78.07%), and apoptosis index (57.5, 43.68, and 65.52%). This study provides scientific evidence for the antimutagenic potential of mangaba fruit pulp and emphasizes its potential as a functional food with widespread applicability in the food industry.

Optimized Camellia sinensis var . sinensis , Ilex paraguariensis , and Aspalathus linearis blend presents high antioxidant and antiproliferative activities in a beverage model

A statistical optimization study was conducted to obtain a tea containing fermented rooibos (Aspalathus linearis), white tea (Camellia sinensis var. sinensis), and roasted mate (Ilex paraguariensis). An optimal combination of these species was proposed. This optimized tea inhibited 64% the lipoperoxidation in vitro and presented a high phenolic content, especially kaempferol, (+)-catechin, (-)-epicatechin, rutin, (-)-epigallocatechin, and (-)-epicatechin-2-O-gallate. Indeed, the antioxidant effect was confirmed by decreasing 30% the reactive oxygen species generation in human hepatoma carcinoma cells (HepG2, 100 and 240 µg/mL). In the cell viability assay, the GI50 for human colorectal adenocarcinoma epithelial cells (Caco-2) was about 547 µg/mL and 481 µg/mL for HepG2. The pasteurization process (65 °C/30 min) did not affect the total phenolic content and antioxidant activity of the optimized tea formulation. The sensory test indicated an acceptability index of 78%, showing that the analytical approach adopted was feasible to develop a phenolic-rich beverage.

In vivo antimutagenic properties of transgenic and conventional soybeans.

The goal of the this study was to evaluate the mutagenic/antimutagenic effects of conventional (BRS133) and transgenic (BRS 245 RR) soybeans (CS and TS, respectively) in vivo using the bone marrow micronucleus (MN) test, histopathological analysis, chromosome aberration test (CAT), and mitotic index (MI) determination. Six-week-old male Swiss mice were fed with pelleted commercial diet mixed with CS or TS at 10% or 20%. Two experimental designs (MN and CAT) were conducted simultaneously with 10 groups each during a 15-day period. Animals were treated with pelleted commercial diet, CS (10% or 20%), or TS (10% or 20%), and on day 14 they also received cyclophosphamide (CP) (50 mg/kg i.p.). The 10% and 20% CS and TS diets did not significantly decrease the frequencies of micronucleated polychromatic erythrocytes in bone marrow induced by CP. However, the CAT indicated that the 10% and 20% CS diets significantly (P < .05) protected nucleated bone marrow cells against chemical-induced mutagenesis and also produced a significant (P < .05) decrease in the total percentage of spontaneous aberrations. Among the treatments with TS, only the 10% TS diet reduced the percentage of total aberrations induced by CP. The results also indicated that the treatment with 20% TS alone significantly (P < .05) decreased the MI, indicating cytotoxic effects related to the treatment. Taken together, our results suggest that, under the tested conditions, TS and CS have antimutagenic properties and are not toxic.

Soja transgênica BRS 243 RR: determinação de macronutrientes e das isoflavonas daidzeína e genisteína por Cromatografia Líquida de Alta Eficiência (CLAE)

The objective of this work was to evaluate the proximate composition, as well as Daidzein (D) and Genistein (G) contents by HPLC, of BRS 243 RR soybean. Sample preparation for the chromatographic analysis involved the use of hexane to remove lipids. Isoflavones were extracted with 70% ethanol containing 0.1% acetic acid. The optimized chromatographic conditions were: C18 column, mobile phase methanol:5% acetic acid (1:1 v/v), flow rate 0.5 mL/minute, column temperature 30 °C, UV absorbance at 254 nm and volume injected 40 µL. The validation parameters were: linearity of daidzein (y = 11242 x -37433, r = 0.9976) and genistein (y = 18510 x -66761, r = 0.9980); variation coefficients obtained in intra-day precision assays [CV = 5.3% (D), CV = 6.7% (G)] and inter-day precision assays [CV = 8.7% (D), CV = 9.7% (G)]; quantification limit 10 µg.g-1; detection limit 5 µg.g-1 and recovery 95.7%. Therefore, the optimized method was appropriate for the determination of daidzein and genistein in soybean. Carbohydrate (31.4%), protein (35.9%), lipid (20.9%), moisture (6.9%), ash (4.9%), daidzein (44.1 µg.g-1) and genistein (37.4 µg.g-1) contents determined in transgenic soybean were similar to those of conventional soybean determined in other studies.

Conventional (MG-BR46 Conquista) and Transgenic (BRS Valiosa RR) Soybeans Have No Mutagenic Effects and May Protect Against Induced-DNA Damage In Vivo

In the present study, we evaluated the pesticide and metal concentrations as well as the antimutagenic and mutagenic properties of commercial soybeans (Glycine max). Male Swiss mice were fed diets containing 1%, 10%, or 20% (w/w) transgenic soybeans (BRS Valiosa RR) or parental isogenic conventional soybeans (MG-BR46 Conquista). Cyclophosphamide (50 mg kg−1 b.w.) was added in a single dose 24 h before euthanasia as an induction agent. There was no difference in the composition (ash, total fat, protein, moisture, and carbohydrates) of the diets containing the same soybean concentration. The results show that the commercially available Brazilian soybeans tested are free of organochlorine, organophosphate, and carbamate pesticides and contain acceptable heavy metal concentrations. Both cyclophosphamide and soybean treatments were not sufficient to cause detectable oxidative damage on liver by the levels of malondialdehyde and protein carbonyl. The transgenic soybeans are also nonmutagenic and have protective effects against DNA damage similar to those of conventional soybeans but to a lesser percentage (64%–101% for conventional and 23%–33% for transgenic diets).

Estimate of iron and vitamin C intake in adolescents during the menstrual cycle phases

The vitamin C and iron ingestion level was evaluated in adolescents during the three phases of the menstrual cycle. The food intake of 30 adolescents was analyzed during the premenstrual, menstrual, and postmenstrual phases, by the registration technique, with the analysis of two registrations per individual, in each phase. The prevalence estimate was done considering six dietary registrations for each adolescent. The patterns used as reference were the Dietary Reference Intakes (DRI) - Estimated Average Requirement (EAR) from Institute of Medicine - IOM (2001), using the probability approaches and the cut - point method for the iron and vitamin C respectively, in addition to the values from the Recommended Dietary Allowances (National Research Council). The adolescents were aged 15.5 +/- 1.7; the age of the menarche was 11.8 +/- 1,1; and gynecological age 3.6 +/- 1.9. The medial consumption of iron was 10.6 mg/day and of vitamin C 77.6 mg/day. It was observed by DRIs-EAR an estimate of inadequacy for iron of 22.6% and for vitamin C of 34% and by National Research Council, 63.3% and 20%, respectively. Despite the divergence among the patterns, both results shows a nutritional risk for the studied population. The accomplishment of wider dietary studies, using DRI-EAR is suggested, favoring nutritional diagnoses with subsequent evaluation of intervention measures.