European Journal of Nutrition (v.51, #2)

Foreword by Stefan Zimmer (11-12).

Introduction: functional foods and oral health by Xiaojie Wang; Adrian Lussi (13-14).

Functional foods/ingredients and dental caries by Cor van Loveren; Zdenek Broukal; Edgar Oganessian (15-25).

Functional foods/ingredients and periodontal diseases by Marja L. Laine; Wim Crielaard (27-30).

Functional foods/ingredients on dental erosion by Xiaojie Wang; Adrian Lussi (39-48).

Numerous supplements containing conjugated linoleic acid (CLA) are presently being promoted for body weight reduction. The aim of this systematic review is to evaluate the evidence for or against the long-term efficacy of CLA.Electronic searches were conducted to identify relevant randomized clinical trials (RCTs). No restrictions in age, time, or language were imposed. Studies had to be at least 6 months in duration. Three reviewers independently determined the eligibility of studies. Two reviewers independently extracted data and assessed the reporting quality of all RCTs.Fifteen RCTs were identified, and seven were included. Four of the included RCTs had serious flaws in the reporting of their methodology. A meta-analysis revealed a statistically significant difference in weight loss favouring CLA over placebo (mean difference: −0.70 kg; 95% confidence interval: −1.09, −0.32). Our meta-analysis also revealed a small significant difference in fat loss favouring CLA over placebo (MD: −1.33 kg; 95% CI: −1.79, −0.86; I 2 = 54%). The magnitude of these effects is small, and the clinical relevance is uncertain. Adverse events included constipation, diarrhea, and soft stools.The evidence from RCTs does not convincingly show that CLA intake generates any clinically relevant effects on body composition on the long term.
Keywords: Obesity; Body weight; Body fat; Weight loss; Fat loss; Meta-analysis

The aim of this study was to evaluate the effect of CLA supplementation on antioxidant metabolism in healthy overweight/obese Korean individuals.We performed a randomized, double-blind, placebo-controlled trial, where 29 healthy overweight/obese (BMI ≥ 23 kg/m2) participants (2 men and 27 women) were randomly selected to receive placebo (n = 15, 2.4 g olive oil/day) or 2.4 g/day CLA mixture (n = 14, 36.9% of cis-9, trans-11 and 37.9% of trans-10, cis-12) for 8 weeks.There were no significant differences in plasma total radical-trapping antioxidant potential (TRAP), lipid peroxidation (conjugated dienes), lipid-soluble antioxidant vitamin concentration, erythrocyte antioxidant enzyme (superoxide dismutase, catalase, glutathione peroxidase) activities, and leukocyte DNA damage between the CLA and placebo groups.The data suggest that short-term supplementation (8 weeks) with CLA (2.4 g/day) might have no significant effects on lipid peroxidation and antioxidant metabolism.
Keywords: Conjugated linoleic acid; Overweight/obese; Antioxidant status

Is vitamin D status known among children living in Northern Italy? by Giuseppina Marrone; Ilaria Rosso; Raffaella Moretti; Francesca Valent; Carla Romanello (143-149).
To assess vitamin D status in children aged 2–220 months in northeastern Italy (latitude 46°). Serum 25-hydroxyvitamin D (25OHD) concentration was assessed in 93 children afferent to the Pediatric Department of the Hospital of Udine.Vitamin D status was defined as follows: sufficient with serum 25OHD between 50 and 250 nmol/l (level 4); insufficient between 37.5 and 50 nmol/l (level 3); deficient less than 37.5 nmol/l (level 2); severely deficient if less than 12.5 nmol/l (level 1). We investigated the potential risk factors of vitamin D deficit.We found that six children (6.4%) had level 1, 36 (38.7%) had level 2, 9 (9.7%) had level 3, and only 45.2% had sufficient level of 25OHD. Immigrate children had a higher risk for vitamin D deficiency if compared with Italians: 75% of non-Italian children had an insufficient 25OHD level compared with 47.0% of Italian children (p = 0.0036). There was a marked seasonal effect on 25OHD level: when plasma sample was withdrawn between November and May, only 29.4% of children showed sufficient vitamin D level, while 70.5% was insufficient (p < 0.0001). Among the obese children, 9.0% had sufficient level of 25OHD with 90% being deficient (p = 0.01). We did not find any significant difference in vitamin D status among children in different age groups.Vitamin D deficiency is common in children living in northeastern Italy. The risk factors were winter season for blood withdrawal, non-Caucasian race, and obesity. These high-risk groups should be targeted for screening and educated about the need of sunlight exposure.
Keywords: Nutrition; Vitamin D; Season; Race

Limited and excess protein intake of pregnant gilts differently affects body composition and cellularity of skeletal muscle and subcutaneous adipose tissue of newborn and weanling piglets by Charlotte Rehfeldt; Louis Lefaucheur; Jana Block; Bernd Stabenow; Ralf Pfuhl; Winfried Otten; Cornelia C. Metges; Claudia Kalbe (151-165).
This study investigated whether dietary protein intake less (50%) or greater (250%) than requirements throughout gestation differently affects offspring body composition and cellular properties of skeletal muscle and subcutaneous adipose tissue (SCAT).Primiparous gilts were fed iso-energetic diets containing adequate (22 AP), high (21 HP), or low (19 LP) protein contents. Newborn (n = 166) and weanling piglets cross-fostered to sows fed a standard diet (day 28; n = 83) were examined by morphological, biochemical, histological, and molecular analyses of the body, SCAT, and semitendinosus, longissimus, biceps femoris muscles.Lowered birth weight (BW) in response to the HP and LP diets (p < 0.01) resulted from decreases in all body constituents in LP, and mainly from reduced body fat in HP piglets (p < 0.05). In the light BW class within litters, HP piglets exhibited a greater percentage of muscle tissue (p < 0.05) than LP piglets. Less SCAT mass in HP and LP piglets resulted from reduced (p < 0.05) number, but not the size of adipocytes. The LP diet adversely affected myogenesis and muscular differentiation derived from less (p < 0.01) primary and secondary myofibers, lower creatine kinase activity (p < 0.05), less IGF2 mRNA (p < 0.10), and greater expression of the embryonic myosin heavy chain isoform (p < 0.01). Catch-up growth of LP but not HP pigs until day 28 increased body fat (p = 0.01). Despite compensated muscle growth in LP piglets, the deficit in myofiber number remained.Poor intrauterine environment by limited and excess protein supply retards fetal growth, but only limited protein supply impairs myogenesis, persistently restricts muscle growth potential, and favors obesity at infancy.
Keywords: Pregnancy protein intake; Pig; Fetal development; Body composition; Muscle fiber; Adipocyte

Identification of change-points in the relationship between food groups in the mediterranean diet and overall mortality: an ‘a posteriori’ approach by Francesco Sofi; Rosanna Abbate; Gian Franco Gensini; Alessandro Casini; Antonia Trichopoulou; Christina Bamia (167-172).
Adherence to Mediterranean diet has been shown to be associated with a better health and greater survival. The aim of the present study was to identify change-points in the relationship between food groups composing Mediterranean diet and overall mortality.The population of the Greek EPIC prospective cohort study (23,349 adult men and women in the Greek EPIC sample who had not previously been diagnosed as having cancer, coronary heart disease or diabetes mellitus at enrolment) was analysed. Segmented logistic regression analysis was conducted to examine the association between each of the food groups contributing to the Mediterranean diet score and overall mortality.This analysis allowed the determination of the following change-points: among men: 1 change-point for vegetables, legumes, cereals, fish and seafood and dairy products and 2 change-points for fruit and nuts, meat and meat products and ethanol; among women: 1 change-point for legumes and fish and seafood and 2 change-points for the remaining food groups. These cut-off points were used to construct an ‘a posteriori’ score that may be better in capturing the health-promoting potential of the traditional Mediterranean diet.Identification of change-points in the relationship between components of the Mediterranean diet and mortality can be used to increase the discriminatory ability of a widely used Mediterranean diet score in relation to mortality.
Keywords: Mediterranean diet; Adherence; Score; Food group

Dietary protein intake in community-dwelling, frail, and institutionalized elderly people: scope for improvement by Michael Tieland; Karin J. Borgonjen-Van den Berg; Luc J. C. van Loon; Lisette C. P. G. M. de Groot (173-179).
Adequate dietary protein intake is required to postpone and treat sarcopenia in elderly people. Insight into dietary protein intake in this heterogeneous population segment is needed to locate dietary inadequacies and to identify target populations and feeding strategies for dietary interventions. Therefore, we assessed dietary protein intake, distribution of protein intake throughout the day, and the use of protein-containing food sources in community-dwelling, frail, and institutionalized elderly people in the Netherlands.Secondary analyses were carried out using dietary data collected from studies among community-dwelling, frail, and institutionalized elderly people to evaluate protein intake characteristics.Dietary protein intake averaged 1.1 ± 0.3 g/kg-bw/day in community-dwelling, 1.0 ± 0.3 g/kg-bw/day in frail, and 0.8 ± 0.3 g/kg-bw/day in institutionalized elderly men. Similar protein intakes were found in women. Ten percent of the community-dwelling and frail elderly and 35% of the institutionalized elderly people showed a protein intake below the estimated average requirement (0.7 g/kg-bw/day). Protein intake was particularly low at breakfast in community-dwelling (10 ± 10 g), frail (8 ± 5 g), and institutionalized elderly people (12 ± 6 g) with bread and dairy products as predominant protein sources.Whereas daily protein intake is generally well above the recommended dietary allowance in community-dwelling and frail elderly people, a significant proportion of institutionalized elderly showed an intake below the current protein requirement, making them an important target population for dietary interventions. Particularly at breakfast, there is scope for improving protein intake.
Keywords: Skeletal muscle mass; Sarcopenia; Frail; Institutionalized; Nutrition; Aging

In vitro activity of dietary flavonol congeners against human cancer cell lines by Chrisiida Tsimplouli; Costas Demetzos; Margarita Hadzopoulou-Cladaras; Panayotis Pantazis; Konstantinos Dimas (181-190).
Flavonoids have physiological activity and a variety of pharmacological properties, including anticancer activity in vitro, but structure–anticancer activity relationships are unclear.The objectives of this work were to investigate the activity of dietary flavonol congeners against cell lines derived from human solid tumours and to examine whether the in vitro activity was associated with specific structural feature(s) of the molecules.Antiproliferative activity of the flavonol congeners was investigated against eight different human cancer cell lines representing different types of human solid tumour, using the sulforhodamine B (SRB) assay in accordance with the instructions published by the NCI. Cell cycle perturbations caused by the congeners were monitored by flow-cytometric analysis of DNA stained with propidium iodide.Most of the flavonols examined had weak antiproliferative and cytotoxic activity. Of all the flavonol congeners tested peracetylated tiliroside found to be the most powerful, with significant antiproliferative and cytotoxic activity. Most flavonols induced similar cell cycle perturbations, whereas induction of apoptosis was significant only for cells treated with peracetylated tiliroside.These findings indicated that the –OH groups of aromatic ring B were not linked to the cytotoxic and antiproliferative activity of the tested flavonols whereas peracetylation of the glycosides resulted in moderate improvement. In contrast, acetylation of tiliroside esterified with coumaric acid at position 5 of the sugar moiety greatly improved the activity of this congener. Overall, the results of this study suggest a critical role of sugar moiety substituents in the anticancer activity of the flavonols.
Keywords: Flavonols; Congeners; Tiliroside; Cytotoxicity; Cell cycle

Diet containing low n-6/n-3 polyunsaturated fatty acids ratio, provided by canola oil, alters body composition and bone quality in young rats by Carlos Alberto Soares da Costa; Aluana Santana Carlos; Gabrielle de Paula Lopes Gonzalez; Rejane Pontes Gaspar Reis; Mariana dos Santos Ribeiro; Aline de Sousa dos Santos; Alexandra Maria Vieira Monteiro; Egberto Gaspar de Moura; Celly Cristina Alves do Nascimento-Saba (191-198).
Adipocytes and osteoblasts were derived from a common progenitor, and canola oil intake may have an adipogenic and osteogenic effect. Thus, our objective was to evaluate the effect on adipocyte, lipid profile, glucose homeostasis, and bone of canola oil as main lipid source on the diet during development.After weaning, rats were divided into two groups (n = 10 per group): control (S) and experimental (C) diets containing 7 mL/100 g soybean or canola oil, respectively. At 60 days, body composition, liver and intra-abdominal fat mass, adipocyte morphology, serum analysis, femur and lumbar vertebras density by dual-energy X-ray absorptiometry and computed tomography were determined. Differences were considered significant with P < 0.05.C group showed the following: lower liver (−12%) and intra-abdominal fat mass (−19%) area of adipocyte (−60%), cholesterol (−33%), insulin (−22%), lower total body (−9%) and spine (−33%) bone mineral content and bone area (−7 and −24%, respectively), femur mass (−9%), width of the diaphysis (−6%), femur (−10%) and lumbar vertebrae bone mineral density (−9%), and radiodensity of femoral head (−8%).The lower intra-abdominal adiposity could have more beneficial effects in a short term, since it can be associated with a better insulin sensitivity and lipid profile, than the small reduction in femur and lumbar vertebra density. However, it has to be considered the incremental effect of this reduction along the aging process.
Keywords: Canola oil; Soybean oil; Adipocyte; Bone; Dual-energy X-ray absorptiometry; Computed tomography; Rats

Influence of gender on DHA synthesis: the response of rat liver to low dietary α-linolenic acid evidences higher ω3 ∆4-desaturation index in females by Jean-Marc Alessandri; Audrey Extier; Kaïs H. Al-Gubory; Emilie Harbeby; Marie-Sylvie Lallemand; Alain Linard; Monique Lavialle; Philippe Guesnet (199-209).
The conversion rate of α-linolenic acid (ALA) into docosahexaenoic acid (DHA) is determined by dietary and non-dietary factors. Higher capacity of DHA synthesis has been evidenced in females, indicating that sex factors influence the conversion pathway. To evaluate the extent to which sexual dimorphism of DHA synthesis is subordinated to nutritional handling, we measured the ω3 ∆4-desaturation index in male and female rats receiving adequate or inadequate amounts of ALA. The ω3 ∆4-desaturation index was drawn from the DHA to docosapentaenoic acid (ω3DPA) ratio in liver phospholipids.Male and female rats born to ω3-deficient dams were fed a supplemented diet supplying low, inadequate, intermediate, or adequate ALA (5, 20, 100, or 300 mg ALA/100 g diet, respectively). Control rats from both gender received the adequate diet from fetal life.Compared with control, low ALA feeding induced the ω3 ∆4-desaturation index to increase by 38 and 70% in the phosphatidylethanolamine fraction of males and females, respectively, and by 67% in phosphatidylcholine in females only. Supplementations with increased doses of ALA progressively smoothed this gender effect. Moreover, the analysis of our data from a previous study shows that ovariectomy decreased, whereas estradiol treatment increased the ω3 index to values comparable with those of diet-matched males and intact females, respectively.Females are more prone than males to increase their index of ω3 ∆4-desaturation, especially in response to low supplies in ALA. Estradiol supports the ω3 index, suggesting that this hormone plays a role in the effect of gender on DHA synthesis.
Keywords: Docosahexaenoic acid; α-Linolenic acid; Desaturation index; Δ6-Desaturase; ω3 Fatty acid; Requirements; Gender effect; Ovariectomy; Estrogen; Liver

Probiotics and dietary counselling targeting maternal dietary fat intake modifies breast milk fatty acids and cytokines by Ulla Hoppu; Erika Isolauri; Päivi Laakso; Jaakko Matomäki; Kirsi Laitinen (211-219).
Breast milk fatty acids possess immunomodulatory properties, and new intervention strategies beyond supplementation of maternal diet with single oils are called for. The objective of the present study was to evaluate the effect of dietary intervention during pregnancy and breastfeeding on breast milk fatty acid and cytokine composition.Pregnant women were randomised into three study groups: dietary intervention with probiotics (diet/probiotic) or with placebo (diet/placebo) and a control group (control/placebo). Dietary intervention included dietary counselling and provision of rapeseed oil-based food products. The probiotics used were Lactobacillus rhamnosus GG and Bifidobacterium lactis Bb12 in combination. Dietary intake was evaluated by food records at every trimester of pregnancy and 1 month postpartum. Breast milk samples were collected after birth (colostrum) and 1 month after delivery for fatty acid and cytokine analysis (n = 125).Dietary intervention improved the quality of fat in the diet. In breast milk, the proportion of α-linolenic acid and total n-3 fatty acids was higher in both dietary intervention groups compared with control group (p < 0.05). In the diet/probiotic group, the γ-linolenic acid content was higher compared with the diet/placebo group (p < 0.05). The concentrations of TNF-α, IL-10, IL-4 and IL-2 were higher in both dietary intervention groups compared with controls, and furthermore, long-chain n-3 fatty acids were associated with several cytokines in colostrum samples.The present intervention demonstrated the possibility of modifying breast milk immunomodulatory factors by dietary means.
Keywords: Breast milk; Fatty acid; Cytokine; Dietary counselling

Iron deficiency anaemia (IDA) is the last stage of iron deficiency, consecutive to an imbalance between iron supply through food intake and iron loss through physiological or pathological processes. As well as by haemoglobin levels, IDA is diagnosed by measuring biomarkers of iron stores. Women are most affected by IDA since their teenage years, as menstruation constitutes a chronic iron loss. Oral supplementation with ferrous sulphate is an effective therapy, but gastrointestinal side effects may impair treatment compliance.The present multicentric randomised controlled trial was designed to assess the non-inferiority of a ferrous sulphate prolonged release formulation called V0355 with the referential ferrous sulphate Ferrograd® in a population of Italian women aged 18–50 years diagnosed for IDA. Three hundred and ninety-nine patients were randomised to receive V0355 (80 mg Fe/day) or Ferrograd® (105 mg Fe/day).After 12 weeks of treatment, the difference in the mean haemoglobin level between the two groups was 0.081 g/dL ([−2.986;1.361], p = 0.54), which confirmed the hypothesis of non-inferiority. All the other biochemical parameters (serum iron, serum ferritin, transferrin, and soluble transferrin receptor) and haematological parameters (erythrocytes count, reticulocytes count, haematocrit, and mean corpuscular volume), as well as patient’s anaemia-related symptoms, were not different between treatment groups throughout the study. Furthermore, the incidence of gastrointestinal adverse events of moderate and severe intensity was significantly lower (p = 0.007) in the V0355 group (5.6%) than in the Ferrograd® group (13.9%).V0355 was as efficient as Ferrograd® in the treatment of anaemia and exhibited a better gastrointestinal tolerance profile.
Keywords: Iron deficiency anaemia; Ferrous sulphate; Gastrointestinal tolerance; Non-inferiority

Effect of the consumption of a fruit and vegetable soup with high in vitro carotenoid bioaccessibility on serum carotenoid concentrations and markers of oxidative stress in young men by Rebeca Martínez-Tomás; Elvira Larqué; Daniel González-Silvera; María Sánchez-Campillo; María Isabel Burgos; Anna Wellner; Soledad Parra; Lucy Bialek; Marie Alminger; Francisca Pérez-Llamas (231-239).
To evaluate the effect of the daily intake of a fruit & vegetable soup with high in vitro bioaccessibility of carotenoids on β-carotene and lycopene serum concentrations.Fourteen healthy young men (24 ± 1 years) received 300 mL/day of a carrot, tomato, and broccoli soup, containing 3.9 mg β-carotene and 4 mg lycopene, for 4 weeks followed by a 4-week washout period. The serum carotenoid response and oxidative markers were analyzed after 3 and 4 weeks of soup consumption and after a 4-week washout.The in vitro bioaccessibility of β-carotene and lycopene was 55 and 43%, respectively, in the soup. Serum β-carotene concentrations were significantly higher than baseline (0.33 ± 0.05 μmol/L) after 3 weeks (0.69 ± 0.06 μmol/L) and 4 weeks (0.78 ± 0.10 μmol/L) of soup consumption (P < 0.001). Serum lycopene was also significantly higher compared with baseline levels (0.26 ± 0.08–0.56 ± 0.04 μmol/L and 0.60 ± 0.04 μmol/L, after 3 and 4 weeks, respectively) (P < 0.001). Although the highest concentration of both carotenoids was found after 4 weeks, the levels were not statistically different from the levels at 3 weeks. A 4-week washout significantly decreased serum carotenoid concentrations, although only β-carotene returned to baseline. Glutathione peroxidase (GPx) increased significantly after soup supplementation compared with baseline, while superoxide dismutase was significantly lower only after 3 weeks. Glutathione reductase, lipid, protein, and DNA oxidative markers remained unchanged.The soup contributed to increasing the concentration of each carotenoid by more than 100% after 3 and 4 weeks of consumption, the maximum increase being observed after 4 weeks. Oxidative markers did not show any variation except for GPx. Serum lycopene half-life was longer than that of β-carotene, which may be important for studies evaluating both carotenoids.
Keywords: β-carotene; Lycopene; Oxidative stress; Glutathione peroxidase

Severe nutritional iron-deficiency anaemia has a negative effect on some bone turnover biomarkers in rats by Javier Díaz-Castro; Mercedes Ramírez López-Frías; Margarita S. Campos; Magdalena López-Frías; María J. M. Alférez; Teresa Nestares; María L. Ojeda; Inmaculada López-Aliaga (241-247).
The role of iron (Fe) in bone formation and disease have not received much attention, a fact that is interesting given the known biochemical role that this mineral has upon collagen maturation together with the high prevalence of Fe-deficiency anaemia worldwide.To investigate the changes in bone formation, resorption and mineral content in developing rats with induced nutritional Fe-deficiency anaemia.Thirty male Wistar rats were divided into two groups, a control group receiving AIN-93G diet with normal-Fe content and an anaemic group receiving AIN-93G diet with low-Fe content for 40 days. Both diets were prepared with an adequate calcium (Ca) and phosphorus (P) content. The most representative serum bone turnover biomarkers and femur and sternum calcium and phosphorus content, together with sternum Fe content were determined in both experimental groups.In anaemic rats, bone matrix formation diminished as revealed by the lower amount of procollagen type I N-terminal propeptide. Bone resorption process increased in Fe deficiency as shown by the increase of serum parathyroid hormone, tartrate-resistant acid phosphatase and levels of degradation products from C-terminal telopeptides of type I collagen released to the serum. In addition, mineralization process was affected by Fe deficiency, because Ca and P content in femur decreased markedly.Fe-deficiency anaemia had a significant impact upon bone, affecting bone mineralization, decreasing the matrix formation and increasing bone resorption, therefore it is of great interest to assess bone status in situation of Fe-deficiency anaemia.
Keywords: Iron-deficiency anaemia; Bone status; Rats

Effect of daily fiber intake on luteinizing hormone levels in reproductive-aged women by Audrey J. Gaskins; Sunni L. Mumford; Jean Wactawski-Wende; Enrique F. Schisterman (249-253).
To evaluate whether the association between fiber intake and LH levels is driven by the association between fiber and estradiol, or whether there is an independent association.A prospective cohort of 259 premenopausal women were followed for up to 2 menstrual cycles. Estrogen and LH were measured ≤8 times per cycle at visits scheduled using fertility monitors. Diet was assessed ≤4 times per cycle by 24-h recall. Linear mixed models on the log scale of hormones were utilized to evaluate the total effects of fiber intake. Inverse probability weights were utilized to estimate the independent effect of fiber on LH levels.In unweighted analyses, we observed a significant, inverse association between fiber intake (in 5 g/day increments) and log LH levels (β, −0.051, 95% confidence interval (CI), −0.100, −0.002). No association was observed in the weighted analyses, after estradiol levels were taken into account (β, −0.016, 95% CI, −0.060, 0.027).The decreased levels of LH associated with high fiber intake were attenuated after taking estradiol levels into account, suggesting that the association between fiber and LH is most likely a consequence of fiber’s impact on estradiol and not due to an independent mechanism.
Keywords: Dietary fiber; Luteinizing hormone; Estradiol; Women

Vitamin D status: sunshine is nice but other factors prevail by Göran Toss; Per Magnusson (255-256).

Treatment for vitamin D deficiency: here and there do not mean everywhere by Pietro Ameri; Marta Bovio; Giovanni Murialdo (257-259).