Potential antiproliferative activity of polyphenol metabolites against human breast cancer cells and their urine excretion pattern in healthy subjects following acute intake of a polyphenol-rich juice of grumixama (Eugenia brasiliensis Lam.).

The bioavailability and metabolism of anthocyanins and ellagitannins following acute intake of grumixama fruit, native Brazilian cherry, by humans, and its in vitro antiproliferative activity against breast cancer cells (MDA-MB-231) were investigated. A single dose of grumixama juice was administered to healthy women (n = 10) and polyphenol metabolites were analyzed in urine and plasma samples collected over 24 h. The majority of the metabolites circulating and excreted in urine were phenolic acids and urolithin conjugates, the gut microbiota catabolites of both classes of polyphenols, respectively. According to pharmacokinetic parameters, the subjects were divided into two distinct groups, high and low urinary metabolite excretors. The pool of polyphenol metabolites found in urine samples showed a significant inhibition of cell proliferation and G2/M cell cycle arrest in MDA-MB-231 cells. Our findings demonstrate the large interindividual variability concerning the polyphenol metabolism, which possibly could reflect in health promotion.

Water extracts of cabbage and kale inhibit ex vivo H(2)O(2)-induced DNA damage but not rat hepatocarcinogenesis.

The chemopreventive potential of water extracts of the Brassica vegetables cabbage and kale was evaluated by administering their aqueous extracts in drinking water ad libitum to Wistar rats submitted to Ito's hepatocarcinogenesis model (CB group and K group, respectively - 14 rats per group). Animals submitted to this same model and treated with water were used as controls (W group - 15 rats). Treatment with the vegetable extracts did not inhibit (P > 0.05) placental glutathione S-transferase-positive preneoplastic lesions (PNL). The number of apoptotic bodies did not differ (P > 0.05) among the experimental groups. Ex vivo hydrogen peroxide treatment of rat livers resulted in lower (P < 0.05) DNA strand breakage in cabbage- (107.6 +/- 7.8 microm) and kale- (110.8 +/- 10.0 microm) treated animals compared with control (120.9 +/- 12.7 microm), as evaluated by the single cell gel (comet) assay. Treatment with cabbage (2 +/- 0.3 microg/g) or kale (4 +/- 0.2 microg/g) resulted in increased (P < 0.05) hepatic lutein concentration compared with control (0.5 +/- 0.07 microg/g). Despite the absence of inhibitory effects of cabbage and kale aqueous extracts on PNL, these Brassica vegetables presented protection against DNA damage, an effect possibly related to increased hepatic lutein concentrations. However, it must be pointed out that the cause-effect relationship between lutein levels and protection is hypothetical and remains to be demonstrated.

Water extracts of cabbage and kale inhibit ex vivo H2O2-induced DNA damage but not rat hepatocarcinogenesis

The chemopreventive potential of water extracts of the Brassica vegetables cabbage and kale was evaluated by administering their aqueous extracts in drinking water ad libitum to Wistar rats submitted to Ito’s hepatocarcinogenesis model (CB group and K group, respectively - 14 rats per group). Animals submitted to this same model and treated with water were used as controls (W group - 15 rats). Treatment with the vegetable extracts did not inhibit (P > 0.05) placental glutathione S-transferase-positive preneoplastic lesions (PNL). The number of apoptotic bodies did not differ (P > 0.05) among the experimental groups. Ex vivo hydrogen peroxide treatment of rat livers resulted in lower (P < 0.05) DNA strand breakage in cabbage- (107.6 ± 7.8 µm) and kale- (110.8 ± 10.0 µm) treated animals compared with control (120.9 ± 12.7 µm), as evaluated by the single cell gel (comet) assay. Treatment with cabbage (2 ± 0.3 µg/g) or kale (4 ± 0.2 µg/g) resulted in increased (P < 0.05) hepatic lutein concentration compared with control (0.5 ± 0.07 µg/g). Despite the absence of inhibitory effects of cabbage and kale aqueous extracts on PNL, these Brassica vegetables presented protection against DNA damage, an effect possibly related to increased hepatic lutein concentrations. However, it must be pointed out that the cause-effect relationship between lutein levels and protection is hypothetical and remains to be demonstrated.

Potencial de fibra alimentar em países ibero-americanos: alimentos, produtos e resíduos / Dietary fiber potential in Iberian-American countries: food, products and residues

The reduced intake of dietary fiber (DF) has been associated with the increase in the incidence of several non-transmissible chronic diseases. Recently many researchers from Iberian-American countries have been studying DF of food and industrial residues in order to improve technological knowledge about concentrates production as well as the development and test of enriched products originated from regional food. The present study aimed to evaluate the regional DF potential based on compilation of data about this nutrient. There is great availability of regional and traditional food with considerable DF amounts as we may find in fruits, vegetables and cereals and legumes. Concentrated sources of DF obtained from different industrial residues were characterized and evaluated making possible their utilization in food enrichment. Cookies added to corn residues (16.8 per cent DF in integral weight) in Colombia, and pasta added to lupine fiber in Chile (11.2 per cent) are examples of tested enriched products. Products like bread with whole wheat (12 per cent), products for especial uses as well as pills of citric DF (55 per cent) and food for enteral diets (1-1.9 per cent) are already available on Cuba market. The dietary fiber contents presented in this work allow us to have an idea of the potential of such nutrient in Iberian-American countries. Such potential has been proved to be considerably representative. It is worth to inform that the complete DF database is available on the CYTED X1.6/CNPq Project Web site (http://www.fcf.usp.br/cytedxi16). It provides data on 817 regional, traditional and not conventional foods, that can be searched and utilized by professional from different areas.

Inhibition of beta-amylase activity, starch degradation and sucrose formation by indole-3-acetic acid during banana ripening.

In order to observe the effect of indole-3-acetic acid (IAA) on carbohydrate metabolism, unripe banana (Musa acuminata AAA, cv. Nanicão) slices were infiltrated with the hormone and left to ripen under controlled conditions. The climacteric respiration burst was reduced by the action of IAA, and starch degradation and sucrose formation were delayed. Sucrose synthase (SuSy; EC 2.4.1.13) and sucrose-phosphate synthase (SPS; EC 2.4.1.14) activities and transcript levels were not affected, indicating that prevention of sucrose accumulation was not related to sucrose-metabolizing enzymes. Impairment of sucrose synthesis could be a consequence of lack of substrate, since starch degradation was inhibited. The increase in activity and transcript level of beta-amylase was delayed, indicating that this enzyme could be important in starch-to-sucrose metabolism in bananas and that it might be, at least partially, controlled at the transcriptional level. This is the first report showing that IAA can delay starch degradation, possibly affecting the activity of hydrolytic enzymes such as beta-amylase (EC 3.2.1.2).

Phosphoglucose isomerase from bananas: partial characterization and relation to main changes in carbohydrate composition during ripening.

Some characteristics of phosphoglucose isomerase (PGI, EC 5.3.1.9) from banana were measured during fruit ripening of three banana cultivars. In banana, PGI was present as two dimeric isoenzymes, named PGI1 and PGI2, which had similar native molecular masses but differed in relation to heat stability and isoelectric point. Total PGI activity showed a distinct two-step change during fruit ripening. Before the climacteric period, PGI activity gradually decreased with the starch content, then its activity began to increase with sucrose accumulation. The ratio of PGI1, and PGI2 was constant, indicating that both enzymes would be involved in starch degradation and sucrose synthesis. PGI activity and changes in carbohydrate composition suggests the existence of some control to fit the requirements of the intense carbon flow from starch to sucrose.

[Trypsin inhibitory activity of beans (Phaseolus vulgaris L.): critical evaluation of methods for determination]. / Atividade inibitória de tripsina do feijão (Phaseolus vulgaris L.): avaliação crítica dos métodos de determinação.

High values of residual trypsin inhibitory activity found for heat-treated beans indicated the occurrence of artifices due to the methodology. A critical evaluation of the methods in use and of possible interferents was performed trying to determine the most adequate way of measuring residual trypsin inhibitory activity. Results showed that extraction conditions, pH of reaction and sample preparation are responsible for significant alterations of the values found, requiring careful standardization. This is the first report about the influence of lyophilization upon residual trypsin inhibitory activity of cooked beans.

Fish oil given by gavage increases lymphocyte proliferation and production of hydrogen peroxide by rat macrophages.

1. The effect of fish oil (FO) administration by gavage (0.4% body weight) on macrophage and lymphocyte function was investigated in young male rats. The results were compared with those obtained by administration of soybean oil (SB) and cocoa butter (CB). 2. Lymphocyte proliferation was markedly increased by FO administration compared with control and other oils. 3. Macrophage phagocytosis capacity was not affected by FO, but it was increased by CB and SB. 4. The oils did not affect the production of O2.- but increased the production of H2O2 in the presence of PMA. 5. The administration of the oils did not markedly affect the activity of antioxidant enzymes in macrophages, except for a decrease in superoxide dismutase activity by FO.

Effect of the administration of fish oil by gavage on activities of antioxidant enzymes of rat lymphoid organs.

1. The effect of administration of fish oil by gavage on catalase (CAT), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities of the lymphoid organs and liver was compared with those of soybean oil and cocoa butter. 2. Fish oil did not affect the activities of SOD and CAT but reduced that of GSH-Px in the spleen. In contrast, cocoa butter reduced the CAT activity in the thymus and liver, and soybean oil decreased CAT activity in the thymus. 3. The content of thiobarbituric acid reactive substances of the lymphoid organs was not modified but was increased in plasma.

Banana sucrose-phosphate synthase gene expression during fruit ripening.

A 952-base pair polymerase-chain-reaction product of sucrose-phosphate synthase (SPS) (EC 2.3.1.14) from banana (Musa acuminata cv. Nanicão) fruit was cloned and used to study expression of the enzyme during development and ripening. The deduced amino acid sequence shows that banana SPS has a high homology with the leaf, tap-root and bean enzymes from other species. Enzyme activity, and mRNA and protein levels point to an increase in SPS expression during ripening. The accumulation of sucrose was correlated to starch degradation and happened 4 d after SPS mRNA and activity had reached their maxima. These results indicate that access to substrate and transcriptional activation with increase in SPS expression might be important regulatory events of sweetening during banana fruit ripening.

Administration of fish oil by gavage increases the activities of hexokinase, glucose-6-phosphate dehydrogenase, and citrate synthase in rat lymphoid organs.

1. The effect of administration of fish oil by gavage on key enzyme activities of glucose metabolism of the thymus, spleen, and mesenteric lymph nodes was investigated. 2. The activities of hexokinase, glucose-6-phosphate dehydrogenase, and citrate synthase in the lymphoid organs were markedly raised due to a daily administration of fish oil by gavage (0.4% of body weight). 3. These findings indicate that the therapeutic utilization of fish oil does affect the metabolism of the lymphoid organs, and possibly immune function; however, the mechanism involved remains to be investigated.

[Evolution of phenolic compounds and condensed tannins (Proanthocyanidins) during the development of bean seeds (Phaseolus vulgaris L.)]. / Evolução dos fenólicos totais e taninos condensados (Proantocianidinas) durante o desenvolvimento das sementes do feijão (Phaseolus vulgaris L.).

The evolution of phenolic compounds and tannins (proanthocyanidins) of bean seeds, Phaseolus vulgaris L., (cultivar aroana 80), from anthesis to maturity (10 to 45 days after anthesis), was investigated. During seed development, phenolic compounds and tannins contents increased by seed unit, until the 31st and 21st day after anthesis respectively, decreasing afterwards. The gradual decrease in catechin and the increase of its polymers (intermediate compounds), as well as the gradual increase in alpha-amylase inhibition capacity were indications that tannins polymerize during seed development.

Purification and Characterization of Two Major Lectins from Araucaria brasiliensis syn. Araucaria angustifolia Seeds (Pinhão).

Two major lectins (lectin I and lectin II) were purified to homogeneity from the seeds of Araucaria brasiliensis (Gymnospermae). The purity of the lectins was confirmed by polyacrylamide gel electrophoresis, isoelectric focusing, and high performance liquid chromatography. They are glycoproteins in nature containing 6.3 and 2.9%, respectively, of neutral sugar and have absorption coefficients of 3.8 and 4.7, respectively, at 280 nanometers. The molecular weights of both lectins obtained by gel filtration on Sephacryl S-400 were equal: 200,000. After dissociation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, molecular weights were 20,000 and 34,000, respectively, for lectin I and lectin II, suggesting they are decameric and hexameric in nature. The amino acid composition of both lectins showed little difference, but both had high amounts of acidic amino acids and lacked methionine in their molecule. The carbohydrate binding specificity of lectins was directed towards mannose, glucose, and their oligomers. High inhibitory activity was also found with thyroglobulin. The erythroagglutinating activity of the lectins was enhanced in the presence of high-molecular-weight substances both at 37 and 4 degrees C. Divalent cations do not appear to be essential for activity. They maintained their agglutinating activity over a broad but different range of pH: 5.5 to 7.5 and 6.5 to 7.5, respectively. Both lectins agglutinated erythrocytes of human ABO blood types equally well.

Effect of oral phytohaemagglutinin intake on cell adaptation in the epithelium of the small intestine of the rat.

The effect of phytohaemagglutinin (PHA) on the adaptation of the proximal jejunal epithelium and on the distal ileal epithelium was studied in rats. The group receiving PHA gained less weight than controls, and the enterocyte population of their jejunal villi, as well as the morphokinetic parameters (length, population, crypt cell production per crypt) of their jejunal and ileal crypts were higher than those of the controls. The proximal lesion caused by PHA (reduction of villus cell populations) stimulates hyperplasia of the crypt-villus unit of the ileal epithelium with the development of adaptation from afar. These adaptations occurred in animals that ingested PHA even in the presence of severe malnutrition.

[Effect of phytohemagglutinins on cell proliferation of the small intestine epithelium of rats]. / Efeito da fitohemaglutinina na proliferação celular do epitélio do intestino delgado de ratos.

Twenty male rats, were divided into two groups of ten animals each: E group, which received solid diet containing 4% of phytohemagglutinin and P group iso-caloric par-fed control, which received the same diet but the phytohemagglutinin was inactivated by heat. Water was offered ad libitum to all groups. The animals were weighed every day and the consumption of diet and water was registered. In the fourteenth day of experiment, the animals were sacrificed and the fragments of jejunum and ileum were removed to morphokinetic study. The results showed that the hydric ingestion was the same in both groups, the body weight of the E group was significant smaller than P group, the villus cell population from the jejunum of the E group was statistically smaller than P group and the contrary happened with ileum samples, wherein the E group was statistically larger than P. The jejunum villus height from E group was similar with P group, but in the ileum of the E group was larger than P. The depth, the cell population and cell production rate of crypt of E group were larger than P, in the jejunum and in the ileum. In conclusion, these results in the present study supply evidence that the intake of the phytohemagglutinin provokes injury of jejunal mucosa, reducing the villus cell population and stimulating the crypt hyperplasia, developing local adaptation. This adaptative model is similar to the one that occurs in celic disease. This proximal lesion stimulate crypt-villus unit hyperplasia of the ileal epithelium, developing distal adaptation. These adaptations occurred in animals that ingested phytohemagglutinin, even though with multicarencial malnutrition.

[Carbohydrates and fiber]. / Considerações sobre carboidratos e fibra.

Dietary carbohydrates comprise two fractions that may be classified as digestible, and which are useful as energy sources (simple and complex carbohydrates) and fiber, which is presumed to be of no use to the human body. There are insufficient epidemiologic data on the metabolic effects of simple carbohydrates and it is not advisable to make quantitative recommendations of intake. It is questionable to recommend in developing countries that a fixed proportion of dietary energy be derived from simple sugars, due to the high prevalence of deficient energy intake, cultural habits, and regional differences in food intake and physical activity. In relation to recommendations of complex carbohydrates, it should be considered that their absorption is influenced by many factors inherent to the individual and to the foods. Fiber is defined as a series of different substances derived from tissue structures, cellular residues and undigested chemical substances that may be partially utilized after intestinal bacteria have acted on them. There is not a clear definition of the chemical composition of fiber, but it consists mainly of polysaccharides (such as cellulose, hemicellulose and pectins), lignin and end products of the interactions of various food components. The effects of fiber, such as control of food intake, regulation of gastrointestinal transit, post-prandial blood concentrations of cholesterol, glucose and insulin, flatulence and alterations in nutrient bioavailability are due to various physical properties inherent to its chemical components. Impairment of nutrient absorption may be harmful, mainly among populations whose food intake is lower than their energy needs, and with a high fiber content. This may be particularly important in pregnant women, growing children and the elderly, and should be considered when making nutrient recommendations. A precise knowledge of fiber is also important to calculate the real energy value of foods, mainly for two reasons: 1) the proportion of "crude fiber" (as measured by acid and alkaline digestion) leads to an over-estimation of the proportion of digestible carbohydrates calculated by difference; 2) fiber may alter the polysaccharide utilization of some foods, as shown by the "glycemic index". It is difficult to make recommendations on dietary fiber due to insufficient data on intake, fiber composition, its physiological effects, and epidemiological studies. However, a preliminary evaluation of the diets from most Latin American countries shows large intakes of vegetable foods and, consequently, an adequate fiber intake may be expected.

Parameters involved in binding of porcine pancreatic alpha-amylase with black bean inhibitor: role of sulfhydryl groups, chloride, calcium, solvent composition and temperature.

The amylase inhibitor of black (kidney) beans (Phaseolus vulgaris; MW 53,000) forms a 1:1 stoichiometric complex with porcine pancreatic alpha-amylase (MW 52,000) at pH 5.40. The single sulfhydryl group of the inhibitor and the two sulfhydryl groups of alpha-amylase are not involved in recognition and binding. Chloride ions, required for activity of alpha-amylase at both pH 5.40 and 6.90, are important for inhibitor--enzyme binding at pH 6.90 but not at pH 5.40. Calcium-free alpha-amylase binds with the inhibitor. An increase in the ionic strength of the solvent increases the rate of binding of the inhibitor with alpha-amylase; a decrease in the dielectric constant decreases the rate of binding; and decreasing the temperature increases the dissociation constant, Kd, of the complex. These data support the hypothesis that hydrophobic interaction is of primary importance in complex formation. The activation energy, Ea, for complex formation was found to be 12.4 kcal/mol at pH 5.40 and 24.2 kcal/mol at pH 6.90. In the presence of the poor substrate, p-nitrophenyl-alpha-D-maltoside, the Ea for complex formation was 4.1 kcal/mol at pH 6.90.

[Antinutritional effect of phytohemagglutinins of Phaseolus vulgaris L]. / Ação anti-nutricional das fito-hemaglutininas de Phaseolus vulgaris,L.

The antinutritional effect caused by the ingestion of lectins from two Brazilian varieties of beans: Rico 23 and Jalo, was studied in rats. The two varieties were selected in a previous screening of toxicity in rats: one of them (Jalo) was lethal, and the other (Rico 23) was not, when injected intra-peritoneally. Different amounts of each one of the lectins were added to casein experimental diets and fed to rats. The amount of protein (casein) also varied from 5% to 20%. The addition to the diet of 1% lectins from the Jalo variety caused a growth depression, as well as a decrease in food efficiency ratio and serum glucose; also, it reduced the maltase and invertase activity of the intestinal mucosa. All these effects appeared when the protein contents in the rations were 5% or 10%. At the 20% level only a depression of the maltase activity was observed. Similar effects were shown by the lectins of the Rico 23 variety, but only when added in a higher (5%) percentage to the diet. The phosphatase and protease activity were not changed by any of the lectins. The inhibitor activity that occurred in vivo was not detected in vitro.