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This study examined the efficacy of a probiotic in reducing depressive symptom severity in people with subthreshold depression. In a double-blind, randomised, placebo-controlled trial, a probiotic (1 × 10^9 live cells per strain: Limosilactobacillus fermentum LF16 (DSM26956), Lacticaseibacillus rhamnosus LR06 (DSM21981), Lactiplantibacillus plantarum LP01 (LMG P-21021) and Bifidobacterium longum 04 (DSM23233)) or placebo was taken daily for 12 weeks. Data were collected at baseline, 6 and 12 weeks including psychological symptom severity (Beck Depression Inventory, BDI; Patient Health Questionnaire, PHQ; Hospital Anxiety Depression Scale, HADS; and Depression Anxiety and Stress Scale, DASS). Biomarkers of glycaemia, inflammation (high-sensitivity C-reactive protein, hs-CRP), antioxidant status (total glutathione (GSH)) and stress (cortisol awakening response, CAR) were also measured. Thirty-nine participants (nineteen probiotic; twenty placebo) were enrolled. There were no significant between-group differences in the examined psychological symptom severity scores, despite certain significant within-group changes observed in both groups from baseline to 6 and/or 12 weeks of follow-up. Regarding biomarkers, the probiotic group showed reduced hs-CRP (-1520; 95 % CI -273·7, -2766·2 ng/dl) and CAR (-0·28; 95 % CI -0·05, -0·51 μg/dl) at 12 weeks, but increased total GSH (3·9; 95 % CI 0·1, 7·5 ng/dl) at 6 weeks, compared with the placebo. The current study reported favourable decreases in depressive symptoms in both groups. Although the within-group changes observed in the probiotic group were supported by favourable inflammatory, antioxidant status and stress biomarker changes compared with the placebo, further research is required to shed more light on the role of gut microbiota modulation on emotional regulation.

Olive oil (OO) polyphenols have been shown to improve HDL anti-atherogenic function, thus demonstrating beneficial effects against cardiovascular risk factors. The aim of the present study was to investigate the effect of extra virgin high polyphenol olive oil (HPOO) v . low polyphenol olive oil (LPOO) on the capacity of HDL to promote cholesterol efflux in healthy adults. In a double-blind, randomised cross-over trial, fifty participants (aged 38·5 (sd 13·9) years, 66 % females) were supplemented with a daily dose (60 ml) of HPOO (320 mg/kg polyphenols) or LPOO (86 mg/kg polyphenols) for 3 weeks. Following a 2-week washout period, participants crossed over to the alternate treatment. Serum HDL-cholesterol efflux capacity, circulating lipids (i.e. total cholesterol, TAG, HDL, LDL) and anthropometrics were measured at baseline and follow-up. No significant between-group differences were observed. Furthermore, no significant changes in HDL-cholesterol efflux were found within either the LPOO and HPOO treatment arms; mean changes were 0·54 % (95 % CI (0·29, 1·37)) and 0·10 % (95 % CI (0·74, 0·94)), respectively. Serum HDL increased significantly after LPOO and HPOO intake by 0·13 mmol/l (95 % CI (0·04, 0·22)) and 0·10 mmol/l (95 % CI (0·02, 0·19)), respectively. A small but significant increase in LDL of 0·14 mmol/l (95 % CI (0·001, 0·28)) was observed following the HPOO intervention. Our results suggest that additional research is warranted to further understand the effect of OO with different phenolic content on mechanisms of cholesterol efflux via different pathways in multi-ethnic populations with diverse diets.

Sugar-sweetened beverage (SSB) consumption has been associated with visceral fat partitioning in adults; however, the underlying mechanisms in childhood remain unclear and warrant exploration. This cross-sectional study aimed to investigate the association between SSB consumption and body fat in children aged 9-13 years and the potential modifying effect of children's sex and serum cortisol levels. A sample of 2665 Greek schoolchildren participated in the 'Healthy Growth Study', and anthropometric, body composition, dietary intake and serum cortisol data were assessed. SSB consumption was defined as low (<1 serving/d), medium (1-2 servings/d) or high (>2 servings/d). We used linear regression models to assess the association between SSB consumption and measures of adiposity and to assess effect modification; models were stratified by sex and tertiles of morning serum cortisol. A significant positive association was observed between high SSB consumption and visceral adipose tissue (VAT) ( β = 1·4, 95 % CI 0·4, 2·3, P = 0·01) but not BMI or BMI z -score. When stratified by sex, the association was observed in boys ( β = 1·8, 95 % CI 0·3, 3·4, P = 0·02) but not in girls. When stratified by cortisol levels, SSB consumption was associated with VAT in children with cortisol levels in the lowest tertile ( β = 2·8, 95 % CI 1·0, 4·6, P < 0·01). These results indicate that increased SSB consumption is associated with visceral adiposity in schoolchildren and this association may be modified by sex and morning serum cortisol. To prevent VAT accumulation and concomitant disease risk, dietary interventions should target SSB consumption during childhood.

Little is known about who would benefit from Internet-based personalised nutrition (PN) interventions. This study aimed to evaluate the characteristics of participants who achieved greatest improvements (i.e. benefit) in diet, adiposity and biomarkers following an Internet-based PN intervention. Adults (n 1607) from seven European countries were recruited into a 6-month, randomised controlled trial (Food4Me) and randomised to receive conventional dietary advice (control) or PN advice. Information on dietary intake, adiposity, physical activity (PA), blood biomarkers and participant characteristics was collected at baseline and month 6. Benefit from the intervention was defined as ≥5 % change in the primary outcome (Healthy Eating Index) and secondary outcomes (waist circumference and BMI, PA, sedentary time and plasma concentrations of cholesterol, carotenoids and omega-3 index) at month 6. For our primary outcome, benefit from the intervention was greater in older participants, women and participants with lower HEI scores at baseline. Benefit was greater for individuals reporting greater self-efficacy for 'sticking to healthful foods' and who 'felt weird if [they] didn't eat healthily'. Participants benefited more if they reported wanting to improve their health and well-being. The characteristics of individuals benefiting did not differ by other demographic, health-related, anthropometric or genotypic characteristics. Findings were similar for secondary outcomes. These findings have implications for the design of more effective future PN intervention studies and for tailored nutritional advice in public health and clinical settings.

Traditionally, personalised nutrition was delivered at an individual level. However, the concept of delivering tailored dietary advice at a group level through the identification of metabotypes or groups of metabolically similar individuals has emerged. Although this approach to personalised nutrition looks promising, further work is needed to examine this concept across a wider population group. Therefore, the objectives of this study are to: (1) identify metabotypes in a European population and (2) develop targeted dietary advice solutions for these metabotypes. Using data from the Food4Me study (n 1607), k-means cluster analysis revealed the presence of three metabolically distinct clusters based on twenty-seven metabolic markers including cholesterol, individual fatty acids and carotenoids. Cluster 2 was identified as a metabolically healthy metabotype as these individuals had the highest Omega-3 Index (6·56 (sd 1·29) %), carotenoids (2·15 (sd 0·71) µm) and lowest total saturated fat levels. On the basis of its fatty acid profile, cluster 1 was characterised as a metabolically unhealthy cluster. Targeted dietary advice solutions were developed per cluster using a decision tree approach. Testing of the approach was performed by comparison with the personalised dietary advice, delivered by nutritionists to Food4Me study participants (n 180). Excellent agreement was observed between the targeted and individualised approaches with an average match of 82 % at the level of delivery of the same dietary message. Future work should ascertain whether this proposed method could be utilised in a healthcare setting, for the rapid and efficient delivery of tailored dietary advice solutions.

The current study was aiming to report the prevalence of suboptimal vitamin D status among schoolchildren in Greece and investigate the role of sex, urbanisation and seasonality on vitamin D status. A sample of 2386 schoolchildren (9-13 years old) from four distinct prefectures was examined. The prevalence of 25-hydroxyvitamin D (25(OH)D) concentration <30 and <50 nmol/l (vitamin D deficiency and insufficiency respectively) was 5·2 and 52·5 %, respectively. Girls had a higher prevalence of 25(OH)D<30 (7·2 v. 3·2 %) and 50 nmol/l (57·0 v. 48·0 %) than boys (P<0·001). The highest prevalence rates of 25(OH)D<30 and 50 nmol/l (9·1 and 73·1 %, respectively) were observed during spring (April to June), whereas the lowest (1·5 and 31·9 %, respectively) during autumn (October to December). The prevalence of 25(OH)D<50 nmol/l was higher in urban/semi-urban than rural regions, particularly during spring months (74·6 v. 47·2 %; P<0·001). Female sex, urban/semi-urban region of residence and spring months were found to increase the likelihood of vitamin D deficiency and insufficiency, with the highest OR observed for spring months (7·47; 95 % CI 3·23, 17·3 and 5·14; 95 % CI 3·84, 6·89 for 25(OH)D<30 and 50 nmol/l respectively). In conclusion, despite the southerly latitude, the prevalence of low vitamin D status among primary schoolchildren in Greece is comparable to or exceeds the prevalence reported among children and adolescents on a European level. Sub-populations at highest risk are girls in urban/semi-urban areas during spring months, thus indicating the need for effective initiatives to support adequate vitamin D status in these population groups.

Individual response to dietary interventions can be highly variable. The phenotypic characteristics of those who will respond positively to personalised dietary advice are largely unknown. The objective of this study was to compare the phenotypic profiles of differential responders to personalised dietary intervention, with a focus on total circulating cholesterol. Subjects from the Food4Me multi-centre study were classified as responders or non-responders to dietary advice on the basis of the change in cholesterol level from baseline to month 6, with lower and upper quartiles defined as responder and non-responder groups, respectively. There were no significant differences between demographic and anthropometric profiles of the groups. Furthermore, with the exception of alcohol, there was no significant difference in reported dietary intake, at baseline. However, there were marked differences in baseline fatty acid profiles. The responder group had significantly higher levels of stearic acid (18 : 0, P=0·034) and lower levels of palmitic acid (16 : 0, P=0·009). Total MUFA (P=0·016) and total PUFA (P=0·008) also differed between the groups. In a step-wise logistic regression model, age, baseline total cholesterol, glucose, five fatty acids and alcohol intakes were selected as factors that successfully discriminated responders from non-responders, with sensitivity of 82 % and specificity of 83 %. The successful delivery of personalised dietary advice may depend on our ability to identify phenotypes that are responsive. The results demonstrate the potential use of metabolic profiles in identifying response to an intervention and could play an important role in the development of precision nutrition.

The interplay between the fat mass- and obesity-associated (FTO) gene variants and diet has been implicated in the development of obesity. The aim of the present analysis was to investigate associations between FTO genotype, dietary intakes and anthropometrics among European adults. Participants in the Food4Me randomised controlled trial were genotyped for FTO genotype (rs9939609) and their dietary intakes, and diet quality scores (Healthy Eating Index and PREDIMED-based Mediterranean diet score) were estimated from FFQ. Relationships between FTO genotype, diet and anthropometrics (weight, waist circumference (WC) and BMI) were evaluated at baseline. European adults with the FTO risk genotype had greater WC (AA v. TT: +1·4 cm; P=0·003) and BMI (+0·9 kg/m2; P=0·001) than individuals with no risk alleles. Subjects with the lowest fried food consumption and two copies of the FTO risk variant had on average 1·4 kg/m2 greater BMI (Ptrend=0·028) and 3·1 cm greater WC (Ptrend=0·045) compared with individuals with no copies of the risk allele and with the lowest fried food consumption. However, there was no evidence of interactions between FTO genotype and dietary intakes on BMI and WC, and thus further research is required to confirm or refute these findings.

An efficient and robust method to measure vitamin D (25-hydroxy vitamin D3 (25(OH)D3) and 25-hydroxy vitamin D2 in dried blood spots (DBS) has been developed and applied in the pan-European multi-centre, internet-based, personalised nutrition intervention study Food4Me. The method includes calibration with blood containing endogenous 25(OH)D3, spotted as DBS and corrected for haematocrit content. The methodology was validated following international standards. The performance characteristics did not reach those of the current gold standard liquid chromatography-MS/MS in plasma for all parameters, but were found to be very suitable for status-level determination under field conditions. DBS sample quality was very high, and 3778 measurements of 25(OH)D3 were obtained from 1465 participants. The study centre and the season within the study centre were very good predictors of 25(OH)D3 levels (P<0·001 for each case). Seasonal effects were modelled by fitting a sine function with a minimum 25(OH)D3 level on 20 January and a maximum on 21 July. The seasonal amplitude varied from centre to centre. The largest difference between winter and summer levels was found in Germany and the smallest in Poland. The model was cross-validated to determine the consistency of the predictions and the performance of the DBS method. The Pearson's correlation between the measured values and the predicted values was r 0·65, and the sd of their differences was 21·2 nmol/l. This includes the analytical variation and the biological variation within subjects. Overall, DBS obtained by unsupervised sampling of the participants at home was a viable methodology for obtaining vitamin D status information in a large nutritional study.

Research suggests that repeatedly offering infants a variety of vegetables during weaning increases vegetable intake and liking. The effect may extend to novel foods. The present study aimed to investigate the impact of advising parents to introduce a variety of single vegetables as first foods on infants' subsequent acceptance of a novel vegetable. Mothers of 4- to 6-month-old infants in the UK, Greece and Portugal were randomised to either an intervention group (n 75), who received guidance on introducing five vegetables (one per d) as first foods repeated over 15 d, or a control group (n 71) who received country-specific 'usual care'. Infant's consumption (g) and liking (maternal and researcher rated) of an unfamiliar vegetable were assessed 1 month post-intervention. Primary analyses were conducted for the full sample with secondary analyses conducted separately by country. No significant effect of the intervention was found for vegetable intake in the three countries combined. However, sub-group analyses showed that UK intervention infants consumed significantly more novel vegetable than control infants (32.8 (SD 23.6) v. 16.5 (sd 12.1) g; P =0.003). UK mothers and researchers rated infants' vegetable liking higher in the intervention than in control condition. In Portugal and Greece, there was no significant intervention effect on infants' vegetable intake or liking. The differing outcome between countries possibly reflects cultural variations in existing weaning practices. However, the UK results suggest in countries where vegetables are not common first foods, advice on introducing a variety of vegetables early in weaning may be beneficial for increasing vegetable acceptance.

The aim of the present study was to examine the associations of obesity, percentage body fat and visceral fat mass with body Fe status in a representative sample of 1493 schoolchildren aged 9-13 years. Anthropometric, body composition, biochemical, clinical (Tanner stage, age of menarche) and dietary intake data were collected. Fe deficiency (ID) was defined as transferrin saturation (TS) < 16 %; and Fe-deficiency anaemia (IDA) as ID with Hb < 120 g/l. Obese boys and girls and those in the highest quartiles of percentage body fat mass had significantly higher levels of serum ferritin (P ≤ 0.05) compared to their normal-weight peers and those in the corresponding lowest quartiles. Similarly, obese boys and girls and those in the highest quartiles of percentage body fat and visceral fat mass had significantly lower levels of TS (P ≤ 0.05) compared to normal-weight children and those in the corresponding lowest quartiles. The prevalence of ID and IDA was significantly higher in boys and girls in the highest quartiles of percentage body fat than in peers in the lowest quartile. Higher quartiles of percentage body fat and visceral fat mass were the main significant predictors of ID in boys, after controlling for other important confounders, with OR of 2.48 (95 % CI, 1.26, 4.88) and 2.12 (95 % CI, 1.07, 4.19), respectively. Similar significant associations were observed for girls. In conclusion, percentage body fat and visceral fat mass were positively associated with ID in both sexes of preadolescents. These associations might be attributed to the chronic inflammation induced by excess adiposity.

The present study was intended to examine whether ponderal index (PI) at birth modifies the effect of the fat mass and obesity associated (FTO) rs9939609 polymorphism on adiposity in European adolescents. A total of 628 adolescents aged 14·4 (se 1·3) years (56·8 % female) were recruited. PI was calculated from parental reports of birth weight and length (kg/m³), and the BMI (kg/m²), body fat percentage and fat mass index (FMI, kg/m²) were calculated. The rs9939609 polymorphism was genotyped and physical activity assessed by accelerometry. Sex, duration of pregnancy, pubertal status, centre and physical activity were used as confounders in all the analyses. The minor A allele of the FTO rs9939609 was significantly associated with higher BMI, body fat percentage and FMI (all P < 0·05) but not with PI. Significant interactions between PI and the rs9939609 polymorphism in terms of body fat percentage (P = 0·002) and FMI (P = 0·017) were detected. However, this polymorphism was only significantly associated with higher BMI, body fat percentage and FMI (all P < 0·05) in adolescents in the lower PI tertile. Indeed, both body fat percentage and FMI were higher in those adolescents in the lower PI tertile carrying the A allele of the FTO rs9939609 polymorphism than in those with the TT genotype (25·0 (se 0·8) v. 22·1 (se 1·0) %, adjusted P = 0·030 and 5·6 (se 0·3) v. 4·6 (se 0·4) kg/m2, P = 0·031, respectively). Our findings suggest that those adolescents born with lower PI could be more vulnerable to the influence of the A risk allele of the FTO polymorphism on total adiposity content.

Low dietary Ca intake and vitamin D insufficiency have been implicated as part of the aetiology leading to osteoporosis. The aim of the present study was to examine the effects of a 30-month dietary intervention that combined supplementation of dairy products fortified with Ca and vitamin D3 and lifestyle and nutrition counselling sessions on bone mineral density (BMD) of postmenopausal women. Sixty-six postmenopausal women (aged 55-65 years) were randomised into a dietary group (DG; n 35), receiving daily and for the first 12 months 1200 mg Ca and 7.5 microg vitamin D3, while for the next 18 months of intervention 1200 mg Ca and 22.5 microg vitamin D3 through fortified dairy products, and a control group (CG; n 31) receiving neither counselling nor dairy products. The DG was found to have more favourable changes in arms (P < 0.001), total spine (P = 0.001) and total body BMD (P < 0.001) compared with the CG. Furthermore, a significant increase was observed for the DG in lumbar spine BMD (0.056; 95 % CI 0.009, 0.103), which was not found to differentiate significantly compared with the change observed in the CG (P = 0.075). In conclusion, the present study showed that intakes of vitamin D of about 22.5 microg/d and of Ca close to the recommended level of 1200 mg from fortified dairy foods for 30 months, with compliance ensured by lifestyle and nutrition counselling sessions, can induce favourable changes in arms, total spine and total body BMD of postmenopausal women.

The aim of the current study was to examine whether calcium supplementation could prevent bone loss in postmenopausal women or more favourable outcomes could be obtained via the consumption of dairy products fortified with calcium and vitamin D3. For this purpose changes in bone mineral density (BMD) at different skeletal sites, assessed by dual-energy X-ray absorptiometry, as well as in quantitative ultrasound (QUS) parameters of the calcaneus over 12 months were estimated. A population of 101 postmenopausal women (55-65 years old) was randomized into a dairy group (DG: n 39), receiving approximately 1200 mg calcium/d and 7.5 microg vitamin D3/d through fortified dairy products; a calcium-supplemented group (CaG: n 26) provided with a calcium supplement of 600 mg/d; and a control group (CG: n 36). Over the intervention period the DG was found to have more favourable changes in pelvis (P=0.040), total spine (P<0.001) and total body BMD (P<0.001) than the other groups. A significant increase was also observed for DG in lumbar spine BMD (2.0%; 95% CI 0.5, 3.5) although it did not differentiate significantly compared to the other groups. No significant differences were observed with respect to the changes in QUS parameters. The current study revealed that recommended intakes of vitamin D3 and calcium via fortified dairy products for 12 months can induce favourable changes in pelvis, total spine and total body BMD in postmenopausal women but not in QUS parameters. No such favourable changes were observed via supplementation of calcium alone.