European Journal of Nutrition (v.51, #3)

Effects of micronutrients on DNA repair by Andrew R. Collins; Amaya Azqueta; Sabine A. S. Langie (261-279).
DNA repair is an essential cellular function, which, by removing DNA damage before it can cause mutations, contributes crucially to the prevention of cancer. Interest in the influence of micronutrients on DNA repair activity is prompted by the possibility that the protective effects of fruits and vegetables might thus be explained. Two approaches to measuring repair—monitoring cellular removal of DNA damage and incubating cell extract with specifically damaged DNA in an in vitro assay—have been applied in cell culture, whole animal studies, and human trials. In addition, there are numerous investigations at the level of expression of DNA repair–related genes.Depending on the pathway studied and the phytochemical or food tested, there are varied reports of stimulation, inhibition or no effect on DNA repair. The clearest findings are from human supplementation trials in which lymphocytes are assessed for their repair capacity ex vivo. Studying cellular repair of strand breaks is complicated by the fact that lymphocytes appear to repair them very slowly. Applying the in vitro repair assay to human lymphocytes has revealed stimulatory effects on repair of oxidised bases by various micronutrients or a fruit- and vegetable-rich diet, while other studies have failed to demonstrate effects.Despite varied results from different studies, it seems clear that micronutrients can influence DNA repair, usually but not always enhancing activity. Different modes of DNA repair are likely to be subject to different regulatory mechanisms. Measures of gene expression tend to be a poor guide to repair activity, and there is no substitute for phenotypic assays.
Keywords: DNA repair; Base excision repair; Nucleotide excision repair; Micronutrients; Antioxidants

Major dietary patterns and cardiovascular risk factors among young Brazilian adults by Maria Teresa A. Olinto; Denise P. Gigante; Bernardo Horta; Vera Silveira; Isabel Oliveira; Walter Willett (281-291).
Diet is one of the most important modifiable risk factors for cardiovascular diseases. The scientific literature has consistently shown the effects of certain diets on health; however, given the variety of cultures and dietary habits across the world, it is likely that much remains to be learned about dietary patterns and health outcomes. We assessed the associations between main dietary patterns and cardiovascular risk factors among 4,202 young Brazilian adults in a cross-sectional analysis.In a principle components analysis, two main dietary patterns were identified: common Brazilian and processed food. As outcomes, we examined body mass index (BMI), waist circumference (WC), systolic blood pressure (SBP), diastolic blood pressure (DBP), total cholesterol, HDL cholesterol (HDL-c), and LDL cholesterol (LDL-c). Means, crude, and adjusted β coefficients and 95% CIs were estimated according to quintiles of dietary patterns.Common Brazilian scores were inversely associated with BMI, WC, LDL-c, HDL-c, and total cholesterol values among men. Among women, inverse association trends were observed with SBP, DBP, LDL-c, HDL-c, and total cholesterol. The processed food pattern was positively associated with LDL-c, HDL-c, total cholesterol, BMI, and WC values among the men. Among the women, the processed food pattern was not significantly associated with cardiovascular risk factors.In conclusion, our findings confirm that diet has an important role on health during early adulthood. The common Brazilian pattern showed generally healthier trends regarding CVD risk factors, but the ultimate effects on risk of risk of disease are unclear because of the inverse relation with HDL-c levels.
Keywords: Cardiovascular disease; Risk factors; Dietary patterns; Young adult

A gluten metabolism study in healthy individuals shows the presence of faecal glutenasic activity by Alberto Caminero; Esther Nistal; Laura Arias; Santiago Vivas; Isabel Comino; Ana Real; Carolina Sousa; José M. Ruiz de Morales; Miguel A. Ferrero; Leandro B. Rodríguez-Aparicio; Javier Casqueiro (293-299).
To study the gluten metabolism in healthy individuals and its effect over the intestinal microbial activity.The faeces of eleven healthy subjects were analysed under 4 diet regimens: their normal gluten diet, a strict gluten-free diet (GFD), a GFD with a supplemental intake of 9 g gluten/day and a GFD with a supplemental intake of 30 g gluten/day. Gluten content, faecal tryptic activity (FTA), short-chain fatty acids (SCFAs) and faecal glutenasic activity (FGA) were analysed in faecal samples.Faecal gluten contents, FTA, SCFAs and FGA varied significantly with different levels of gluten intake in the diet. When high gluten doses (30 g/day) were administered in the diet, SCFA concentrations (70.5 mmoles/kg faeces) were significantly different from those from the GFD period (33.8 mmoles/kg faeces) of the experiment. However, the FTA showed significant differences between the GFD (34 units) and the normal gluten-containing diet (60 units) and also between the GFD and the GFD + 30 g of gluten/day (67 units). When gluten was present in the diet, gluten was detected in the faeces, showing that at least a portion of the gluten ingested is eliminated in the large intestine, providing a substrate for intestinal microbial proteases. We have also shown the presence of faecal glutenasic activity that increased proportionally with the gluten intake in the diet, showing an enzymatic activity of 993 units in DSG, 2,063 units in DSG + 9 g and 6,090 units in DSG + 30 g.The activity of the intestinal microbiota is modified by gluten intake in the diet. The incorporation of gluten in the diet increases the activity of a gluten proteolytic activity in the faeces.
Keywords: Gluten; Coeliac disease; Faecal glutenasic activity; Gut microbiota

Current evidence suggests that consumption of virgin olive oil (VOO) helps to protect against the development of atherosclerosis and that minor components such as oleanolic acid contribute to this effect. In this study, the effects of triacylglycerol-rich lipoproteins (TRLs) derived from olive oil on inflammatory processes in macrophages and how they are modulated by oleanolic acid was investigated.TRLs isolated from healthy volunteers 2 and 4 h after a test meal containing VOO, pomace olive oil (POO) (the second pressing of olive oil, enriched in minor components) or POO enriched with oleanolic acid (OPOO) were incubated with macrophages derived from the human monocyte cell line, THP-1.All types of TRLs caused a decrease of about 50% in the secretion of monocyte chemoattractant protein-1 (MCP-1) by the cells. Interleukin (IL)-6 secretion was also significantly decreased by 2 and 4 h VOO TRLs and by 4 h OPOO TRLs. In contrast, increased IL-1β secretion was observed with all 2 h TRL types, and increased tumour necrosis factor-α (TNF-α) production with 2 h VOO and POO, but not OPOO, TRLs. TRLs isolated after 4 h, however, had no significant effects on TNF-α secretion and increased IL-1β secretion only when they were derived from VOO. Cyclooxygenase-2 (COX-2) mRNA expression was strongly down-regulated by all types of TRLs, but protein expression was significantly depressed only by 4 h OPOO TRLs.These findings demonstrate that TRLs derived from olive oil influence inflammatory processes in macrophages and suggest that oleanolic acid may have beneficial effects.
Keywords: Triacylglycerol-rich lipoproteins; Olive oil; Oleanolic acid; Cytokine secretion; Cyclooxygenase-2; Macrophages

Plant polyphenols attenuate hepatic injury after hemorrhage/resuscitation by inhibition of apoptosis, oxidative stress, and inflammation via NF-kappaB in rats by Borna Relja; Eva Töttel; Lara Breig; Dirk Henrich; Heinz Schneider; Ingo Marzi; Mark Lehnert (311-321).
Oxidative stress and inflammation contribute to hepatic injury after hemorrhage/resuscitation (H/R). Natural plant polyphenols, i.e., green tea extract (GTE) possess high anti-oxidant and anti-inflammatory activities in various models of acute inflammation. However, possible protective effects and feasible mechanisms by which plant polyphenols modulate pro-inflammatory, apoptotic, and oxidant signaling after H/R in the liver remain unknown. Therefore, we investigated the effects of GTE and its impact on the activation of NF-kappaB in the pathogenesis of hepatic injury induced by H/R.Twenty-four female LEWIS rats (180–250 g) were fed a standard chow (ctrl) or a diet containing 0.1% polyphenolic extracts (GTE) from Camellia sinensis starting 5 days before H/R. Rats were hemorrhaged to a mean arterial pressure of 30 ± 2 mmHg for 60 min and resuscitated (H/R and GTE H/R groups). Control groups (sham, ctrl, and GTE) underwent surgical procedures without H/R. Two hours after resuscitation, tissues were harvested.Plasma alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) increased 3.5-fold and fourfold, respectively, in vehicle-treated rats as compared to GTE-fed rats. Histopathological analysis revealed significantly decreased hepatic necrosis and apoptosis in GTE-fed rats after H/R. Real-time PCR showed that GTE diminished gene expression of pro-apoptotic caspase-8 and Bax, while anti-apoptotic Bcl-2 was increased after H/R. Hepatic oxidative (4-hydroxynonenal) and nitrosative (3-nitrotyrosine) stress as well as systemic IL-6 level and hepatic IL-6 mRNA were markedly reduced in GTE-fed rats compared with controls after H/R. Plant polyphenols also decreased the activation of both JNK and NFκB.Taken together, GTE application blunts hepatic damage, apoptotic, oxidative, and pro-inflammatory changes after H/R. These results underline the important roles of JNK and NF-kappaB in inflammatory processes after H/R and the beneficial impact of plant polyphenols in preventing their activation.
Keywords: Plant polyphenols; Green tea extract; NF-kappaB; Hemorrhagic shock; Liver; Apoptosis; In vivo

To study the protective effects and underlying molecular mechanisms of SAMC on carbon tetrachloride (CCl4)-induced acute hepatotoxicity in the mouse model.Mice were intraperitoneally injected with CCl4 (50 μl/kg; single dose) to induce acute hepatotoxicity with or without a 2-h pre-treatment of SAMC intraperitoneal injection (200 mg/kg; single dose). After 8 h, the blood serum and liver samples of mice were collected and subjected to measurements of histological and molecular parameters of hepatotoxicity.SAMC reduced CCl4-triggered cellular necrosis and inflammation in the liver under histological analysis. Since co-treatment of SAMC and CCl4 enhanced the expressions of antioxidant enzymes, reduced the nitric oxide (NO)-dependent oxidative stress, and inhibited lipid peroxidation induced by CCl4. SAMC played an essential antioxidative role during CCl4-induced hepatotoxicity. Administration of SAMC also ameliorated hepatic inflammation induced by CCl4 via inhibiting the activity of NF-κB subunits p50 and p65, thus reducing the expressions of pro-inflammatory cytokines, mediators, and chemokines, as well as promoting pro-regenerative factors at both transcriptional and translational levels.Our results indicate that SAMC mitigates cellular damage, oxidative stress, and inflammation in CCl4-induced acute hepatotoxicity mouse model through regulation of NF-κB. Garlic or garlic derivatives may therefore be a potential food supplement in the prevention of liver damage.
Keywords: Liver injury; S-allylmercaptocysteine; Carbon tetrachloride; Oxidative stress; Necroinflammation; Nuclear factor κB

Differential effects of 1α,25-dihydroxycholecalciferol on MCP-1 and adiponectin production in human white adipocytes by Silvia Lorente-Cebrián; Anna Eriksson; Thomas Dunlop; Niklas Mejhert; Ingrid Dahlman; Gaby Åström; Eva Sjölin; Kerstin Wåhlén; Carsten Carlberg; Jurga Laurencikiene; Per Hedén; Peter Arner; Mikael Rydén (335-342).
Obesity is characterized by a low-grade inflammation in white adipose tissue (WAT), which promotes insulin resistance. Low serum levels of 1α,25-dihydroxycholecalciferol (DHCC) associate with insulin resistance and higher body mass index although it is unclear whether vitamin D supplementation improves insulin sensitivity. We investigated the effects of DHCC on adipokine gene expression and secretion in adipocytes focusing on two key factors with pro-inflammatory [monocyte chemoattractant protein-1 (MCP-1/CCL2)] and anti-inflammatory [adiponectin (ADIPOQ)] effects.Pre-adipocytes were isolated from human subcutaneous WAT and cultured until full differentiation. Differentiated adipocytes were either pre-treated with DHCC (10−7 M) and subsequently incubated with tumor necrosis factor-α (TNFα, 100 ng/mL) or concomitantly incubated with TNFα/DHCC. MCP1 and adiponectin mRNA expression was measured by RT–PCR and protein release by ELISA.DHCC was not toxic and did not affect adipocyte morphology or the mRNA levels of adipocyte-specific genes. TNFα induced a significant increase in CCL2 mRNA and protein secretion, while DHCC alone reduced CCL2 mRNA expression (~25%, p < 0.05). DHCC attenuated TNFα-induced CCL2 mRNA expression in both pre-incubation (~15%, p < 0.05) and concomitant (~60%, p < 0.01) treatments. TNFα reduced ADIPOQ mRNA (~80%) and secretion (~35%). DHCC alone decreased adiponectin secretion to a similar degree (~35%, p < 0.05). Concomitant treatment with DHCC/TNFα for 48 h had an additive effect, resulting in a pronounced reduction in adiponectin secretion (~70%).DHCC attenuates MCP-1 and adiponectin production in human adipocytes, thereby reducing the expression of both pro- and anti-inflammatory factors. These effects may explain the difficulties so far in determining the role of DHCC in insulin sensitivity and obesity in humans.
Keywords: Adipocyte; 1α,25-dihydroxycholecalciferol; MCP-1; Adiponectin; Insulin resistance; Obesity

Orally administered bovine lactoferrin (bLF) exerts an anti-inflammatory effect on hepatitis and colitis animal models. To investigate the mechanism underlying the action of bLF, we explored the expression of inflammation-related factors in the intestine of a hepatitis mouse model after the oral administration of bLF and in several human intestinal cell lines treated with bLF.The effects of bLF on the expression of interleukin-11 (IL-11) and bone morphogenetic protein 2 (BMP2) in the intestinal mucosa of a hepatitis mouse model as well as in cell cultures of human intestinal epithelial cells, myofibroblasts, and monocytes were examined using the real-time reverse transcription polymerase chain reaction. Epithelial cells and myofibroblasts were also cocultured using transwells. bLF transport, and IL-11 and BMP2 induction, as well as the interactions between the two cell types, were then analyzed after bLF treatment.In vivo, oral bLF administration increased the production of IL-11 and BMP2 in intestinal specimens. In vitro, bLF only stimulated the production of IL-11 in human intestinal myofibroblasts; i.e., it had no effect on BMP2 production in any cell type. In the transwell cocultures, bLF passed through the epithelium and directly stimulated IL-11 production in the myofibroblasts on the basolateral side. The IL-11 produced in the myofibroblasts subsequently acted protectively on the epithelial cells of the coculture.bLF upregulated the activity of anti-inflammatory factors, such as IL-11, in the intestine of a hepatitis mouse model and human intestinal myofibroblasts.
Keywords: Lactoferrin; Interleukin-11; Bone morphogenetic protein 2; Anti-inflammatory factor

Positive effects of pistachio nut consumption on plasma inflammatory biomarkers have been described; however, little is known about molecular events associated with these effects.We studied the anti-inflammatory activity of a hydrophilic extract from Sicilian Pistacia L. (HPE) in a macrophage model and investigated bioactive components relevant to the observed effects.HPE oligomer/polymer proanthocyanidin fractions were isolated by adsorbance chromatography, and components quantified as anthocyanidins after acidic hydrolysis. Isoflavones were measured by gradient elution HPLC analysis. RAW 264.7 murine macrophages were pre-incubated with either HPE (1- to 20-mg fresh nut equivalents) or its isolated components for 1 h, then washed before stimulating with lipopolysaccharide (LPS) for 24 h. Cell viability and parameters associated with Nuclear Factor-κB (NF-κB) activation were assayed according to established methods including ELISA, Western blot, or cytofluorimetric analysis.HPE suppressed nitric oxide (NO) and tumor necrosis factor-α (TNF-α) production and inducible NO-synthase levels dose dependently, whereas inhibited prostaglandin E2 (PGE2) release and decreased cyclo-oxygenase-2 content, the lower the HPE amount the higher the effect. Cytotoxic effects were not observed. HPE also caused a dose-dependent decrease in intracellular reactive oxygen species and interfered with the NF-κB activation. Polymeric proanthocyanidins, but not isoflavones, at a concentration comparable with their content in HPE, inhibited NO, PGE2, and TNF-α formation, as well as activation of IκB-α. Oligomeric proanthocyanidins showed only minor effects.Our results provide molecular evidence of anti-inflammatory activity of pistachio nut and indicate polymeric proanthocyanidins as the bioactive components. The mechanism may involve the redox-sensitive transcription factor NF-κB. Potential effects associated with pistachio nut consumption are discussed in terms of the proanthocyanidin bioavailability.
Keywords: Inflammation; Isoflavones; Macrophages; Nut; Proanthocyanidins; Sicilian pistachio

The immunomodulatory properties of viable Lactobacillus salivarius ssp. salivarius CECT5713 are not restricted to the large intestine by Belén Arribas; Natividad Garrido-Mesa; Laura Perán; Desirée Camuesco; Mònica Comalada; Elvira Bailón; Mónica Olivares; Jordi Xaus; Laurens Kruidenier; Ian R. Sanderson; Antonio Zarzuelo; Maria Elena Rodríguez-Cabezas; Julio Gálvez (365-374).
The aim of this study was to better characterise the biological effects of Lactobacillus salivarius ssp. salivarius CECT5713, a probiotic with immunomodulatory properties.Live or dead probiotic was assayed in the TNBS model of rat colitis to determine whether viability was a requisite to exert the beneficial effects. In vitro studies were also performed in Caco-2 cells to evaluate its effects on epithelial cell recovery and IL-8 production. Finally, the probiotic was assayed in the LPS model of septic shock in mice to establish its effects when there is an altered systemic immune response.The viability of the probiotic was required for its anti-inflammatory activity. The probiotic inhibited IL-8 production in stimulated Caco-2 cells and facilitated the recovery of damaged intestinal epithelium. In LPS-treated mice, the probiotic inhibited the production of TNFα in plasma and lungs and increased the hepatic glutathione content. These effects were associated with an improvement in the altered production of the T-cell cytokines in splenocytes, by reducing IL-2 and IL-5 and by increasing IL-10. Finally, it reduced the increased plasma IgG production in LPS-treated mice.The anti-inflammatory effects of viable L. salivarius ssp. salivarius CECT5713 are not restricted to the gastrointestinal tract.
Keywords: L. salivarius ssp. salivarius CECT5713; Mice LPS septic shock; Cytokines; Immunoglobulin; Intestinal anti-inflammatory activity; TNBS rat colitis; Caco-2 cells

Effects of creatine in a rat intestinal model of ischemia/reperfusion injury by M. N. Orsenigo; C. Porta; C. Sironi; U. Laforenza; G. Meyer; M. Tosco (375-384).
Creatine belongs to a buffering system of cellular ATP level and has been reported to display direct antioxidant activity. Aim of this work was to investigate whether creatine treatment could ameliorate the antioxidant response of intestinal cells and limit the oxidative injury induced by anoxia and subsequent reoxygenation.Jejunal and ileal tracts of rat intestine were everted and incubated in vitro under normoxic, anoxic and reoxygenation conditions in the absence and in the presence of 10 mM creatine. (Na+, K+)-ATPase, γ-GT and antioxidant enzymes activities were determined in mucosal homogenate, as well as malondialdehyde production and HSP70 expression.Both in jejunum and ileum, creatine treatment increases (Na+, K+)-ATPase activity; γ-GT is unaffected in jejunum but stimulated in ileum. In both tissues, creatine does not alter the antioxidant activities or malondialdehyde level. HSP70 expression is increased only in jejunum. Anoxic conditions stimulate antioxidant activities to a greater extent in jejunum compared to ileum; reoxygenation does not evoke further effects, but enhances malondialdehyde production in both tracts. The protective action of creatine, in reoxygenation, is more marked in jejunum as for its stimulation of antioxidant activities; however, in jejunum, a prooxidant action of creatine is suggested, since malondialdehyde production is enhanced by its presence; on the contrary in ileum, where HSP70 is overexpressed in reoxygenation, peroxidation level is significantly reduced.The presence of creatine seems to potentiate the defensive response of both tissues, in jejunum by means of cell antioxidant equipment, in ileum by the involvement of HSP70.
Keywords: Creatine; Rat; Jejunum; Ileum; Ischemia/reperfusion

Influence of maternal cigarette smoking during pregnancy on neonatal serum folate levels by Mehmet Yekta Oncel; Ramazan Ozdemir; Omer Erdeve; Ugur Dilmen (385-387).
Folate is an essential micronutrient for fetal development because of its role in de novo synthesis of DNA. The aim of this study was to compare neonatal serum folate levels of babies born to smoking and non-smoking mothers.Infants of consenting pregnant mothers presenting at ≥37 weeks of gestation were enrolled. Subjects were divided into two groups based on their mother’s smoking habits. Blood samples were obtained at birth (from the umbilical cord) and 1 month after delivery for the determination of serum folate levels using a chemiluminescence method.Among 140 consenting subjects, 108 (77%) brought their newborns to their scheduled visit 1 month after delivery, 68 of whom were non-smokers and 40 were smokers. Babies born to smoking mothers had significantly lower serum folate levels compared to those born to non-smoking mothers, both at birth (17.2 ± 5 vs. 24.3 ± 4.9; p < 0.01) and 1 month after delivery (11 ± 4.1 vs. 17.5 ± 4.3; p < 0.01).Our study is the first of its kind to demonstrate that smoking results in significant reductions in serum folate levels of newborns. These results suggest that folic acid supplementation may be required for expectant smoking mothers throughout pregnancy, not just during the first trimester. Similar supplementation may also be warranted for infants born to such mothers.
Keywords: Folic acid; Newborn; Smoking