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1.  Lipase-catalyzed biodiesel production and quality with Jatropha curcas oil: exploring its potential for Central America 
Background
Extensive native Jatropha curcas L. (Jatropha) crop areas have been planted in Central America marginal lands since 2008 as a non-edible prospective feedstock alternative to high-value, edible palm oil. Jatropha biodiesel is currently exclusively produced in the region at commercial scale utilizing alkaline catalysts. Recently, a free, soluble Thermomyces lanuginosus (TL) 1,3 specific lipase has shown promise as biocatalyst, reportedly yielding up to 96 % ASTM D6751 compliant biodiesel after 24 h transesterification of soybean, canola oils and other feedstocks. Biodiesel conversion rate and quality of enzymatically catalyzed transesterification of Jatropha oil was evaluated. Two lipases: free, soluble TL and immobilized Candida antarctica (CA) catalyzed methanolic transesterification of crude Jatropha and refined palm oil.
Results
Jatropha yields were similar to palm biodiesel with NaOH as catalyst. After 24 h transesterification, Jatropha (81 %) and palm oil (86 %) biodiesel yields with TL as catalyst were significantly higher than CA (<70 %) but inferior to NaOH (>90 %). Enzymatic catalysts (TL and CA) produced Jatropha biodiesel with optimum flow properties but did not complied with ASTM D6751 stability parameters (free fatty acid content and oil stability index).
Conclusions
Biodiesel production with filtered, degummed, low FFA Jatropha oil using a free liquid lipase (TL) as catalyst showed higher yielding potential than immobilized CA lipase as substitute of RBD palm oil with alkaline catalyst. However, Jatropha enzymatic biodiesel yield and stability were inferior to alkaline catalyzed biodiesel and not in compliance with international quality standards. Lower quality due to incomplete alcoholysis and esterification, potential added costs due to need of more than 24 h to achieve comparable biodiesel yields and extra post-transesterification refining reactions are among the remaining drawbacks for the environmentally friendlier enzymatic catalysis of crude Jatropha oil to become an economically viable alternative to chemical catalysis.
doi:10.1186/s13036-015-0009-9
PMCID: PMC4513677  PMID: 26213567
Biodiesel; Crude oil; Jatropha curcas; Transesterification; Soluble lipase; ASTM D675
2.  Total fluid intake of children and adolescents: cross-sectional surveys in 13 countries worldwide 
European Journal of Nutrition  2015;54(Suppl 2):57-67.
Purpose
To describe total fluid intake (TFI) according to socio-demographic characteristics in children and adolescents worldwide.
Methods
Data of 3611 children (4–9 years) and 8109 adolescents (10–18 years) were retrieved from 13 cross-sectional surveys (47 % males). In three countries, school classes were randomly recruited with stratified cluster sampling design. In the other countries, participants were randomly recruited based on a quota method. TFI (drinking water and beverages of all kinds) was obtained with a fluid-specific record over 7 consecutive days. Adequacy was assessed by comparing TFI to 80 % of adequate intake (AI) for total water intake set by European Food Safety Authority. Data on height, weight and socio-economic level were collected in most countries.
Results
The mean (SD) TFI ranged from [1.32 (0.68)] to [1.35 (0.71)] L/day. Non-adherence to AIs for fluids ranged from 10 % (Uruguay) to >90 % (Belgium). Females were more likely to meet the AIs for fluids than males (4–9 years: 28 %, OR 0.72, p = 0.002; 10–18 years: 20 %, OR 0.80, p = 0.001), while adolescents were less likely to meet the AI than children (OR 1.645, p < 0.001 in males and OR 1.625, p < 0.001 in females).
Conclusions
A high proportion of children and adolescents are at risk of an inadequate fluid intake. This risk is especially high in males and adolescents when compared with females or children categories. This highlights water intake among young populations as an issue of global concern.
Electronic supplementary material
The online version of this article (doi:10.1007/s00394-015-0946-6) contains supplementary material, which is available to authorized users.
doi:10.1007/s00394-015-0946-6
PMCID: PMC4473088  PMID: 26081646
Fluid intake; Children; Adolescents; Worldwide; Dietary assessment; Hydration
3.  Intake of water and beverages of children and adolescents in 13 countries 
European Journal of Nutrition  2015;54(Suppl 2):69-79.
Purpose
To describe the intake of water and all other beverages in children and adolescents in 13 countries of three continents.
Methods
Data of 3611 children (4–9 years) and 8109 adolescents (10–17 years) were retrieved from 13 cross-sectional surveys (47 % males). In three countries, stratified cluster sampling design was applied to randomly recruit schools classes. A quota method was applied in the other countries to randomly recruit participants. Details on the intake of all fluid types were obtained with a fluid-specific record over 7 consecutive days.
Results
In the total sample, the highest mean intakes were observed for water (738 ± 567 mL/day), followed by milk (212 ± 209 mL/day), regular soft beverages (RSB) (168 ± 290 mL/day) and juices (128 ± 228 mL/day). Patterns characterized by a high contribution of water, RSB or hot beverages to total fluid intake were identified among the countries with close geographical location. Adolescents had a significantly lower milk intake and higher intake of RSB and hot beverages than children in most countries. The most consistent gender difference observed was that in both age groups males reported a significantly higher RSB consumption than females.
Conclusion
On average, water was the fluid consumed in the largest volume by children and adolescents, but the intake of the different fluid types varied substantially between countries. Since the RSB intake was as large, or even larger, than water intake in some countries, undertaking actions to improve fluid intake habits of children and adolescents are warranted.
Electronic supplementary material
The online version of this article (doi:10.1007/s00394-015-0955-5) contains supplementary material, which is available to authorized users.
doi:10.1007/s00394-015-0955-5
PMCID: PMC4473084  PMID: 26072216
Water; Beverages; Fluid intake; Children; Adolescents
4.  Resting Heart Rate Is Not a Good Predictor of a Clustered Cardiovascular Risk Score in Adolescents: The HELENA Study 
PLoS ONE  2015;10(5):e0127530.
Background
Resting heart rate (RHR) reflects sympathetic nerve activity a significant association between RHR and all-cause and cardiovascular mortality has been reported in some epidemiologic studies.
Methods
To analyze the predictive power and accuracy of RHR as a screening measure for individual and clustered cardiovascular risk in adolescents. The study comprised 769 European adolescents (376 boys) participating in the HELENA cross-sectional study (2006–2008) were included in this study. Measurements on systolic blood pressure, HOMA index, triglycerides, TC/HDL-c, VO2máx and the sum of four skinfolds were obtained, and a clustered cardiovascular disease (CVD) risk index was computed. The receiver operating characteristics curve was applied to calculate the power and accuracy of RHR to predict individual and clustered CVD risk factors.
Results
RHR showed low accuracy for screening CVD risk factors in both sexes (range 38.5%–54.4% in boys and 45.5%–54.3% in girls). Low specificity’s (15.6%–19.7% in boys; 18.1%–20.0% in girls) were also found. Nevertheless, the sensitivities were moderate-to-high (61.4%–89.1% in boys; 72.9%–90.3% in girls).
Conclusion
RHR is a poor predictor of individual CVD risk factors and of clustered CVD and the estimates based on RHR are not accurate. The use of RHR as an indicator of CVD risk in adolescents may produce a biased screening of cardiovascular health in both sexes.
doi:10.1371/journal.pone.0127530
PMCID: PMC4444318  PMID: 26010248
5.  Differences in Energy Balance-Related Behaviours in European Preschool Children: The ToyBox-Study 
PLoS ONE  2015;10(3):e0118303.
Background
The aim of the current study was to compare levels of energy balance-related behaviours (physical activity, sedentary behaviour, and dietary behaviours (more specifically water consumption, sugar-sweetened beverage consumption and unhealthy snacking)) in four- to six-year-old preschoolers from six European countries (Belgium, Bulgaria, Germany, Greece, Poland, and Spain) within the ToyBox cross-sectional study.
Methods
A sample of 4,045 preschoolers (4.77 ± 0.43 years; 52.2% boys) had valid physical activity data (steps per day), parents of 8,117 preschoolers (4.78 ± 0.46 years; 53.0% boys) completed a parental questionnaire with questions on sedentary behaviours (television viewing, computer use, and quiet play), and parents of 7,244 preschoolers (4.77 ± 0.44 years; 52.0% boys) completed a food frequency questionnaire with questions on water consumption, sugar-sweetened beverage consumption and unhealthy snacking.
Results
The highest levels of physical activity were found in Spain (12,669 steps/day on weekdays), while the lowest levels were found in Bulgaria and Greece (9,777 and 9,656 steps/day on weekdays, respectively). German preschoolers spent the least amount of time in television viewing (43.3 min/day on weekdays), while Greek preschoolers spent the most time in television viewing (88.5 min/day on weekdays). A considerable amount of time was spent in quiet play in all countries, with the highest levels in Poland (104.9 min/day on weekdays), and the lowest levels in Spain (60.4 min/day on weekdays). Belgian, German, and Polish preschoolers had the lowest intakes of water and the highest intakes of sugar-sweetened beverages. The intake of snacks was the highest in Belgian preschoolers (73.1 g/day) and the lowest in Greek preschoolers (53.3 g/day).
Conclusions
Across six European countries, differences in preschoolers’ energy balance-related behaviours were found. Future interventions should target European preschoolers’ energy balance-related behaviours simultaneously, but should apply country-specific adaptations.
doi:10.1371/journal.pone.0118303
PMCID: PMC4364763  PMID: 25785738
6.  Dietary animal and plant protein intakes and their associations with obesity and cardio-metabolic indicators in European adolescents: the HELENA cross-sectional study 
Nutrition Journal  2015;14:10.
Background
Previous studies suggest that dietary protein might play a beneficial role in combating obesity and its related chronic diseases. Total, animal and plant protein intakes and their associations with anthropometry and serum biomarkers in European adolescents using one standardised methodology across European countries are not well documented.
Objectives
To evaluate total, animal and plant protein intakes in European adolescents stratified by gender and age, and to investigate their associations with cardio-metabolic indicators (anthropometry and biomarkers).
Methods
The current analysis included 1804 randomly selected adolescents participating in the HELENA study (conducted in 2006–2007) aged 12.5-17.5 y (47% males) who completed two non-consecutive computerised 24-h dietary recalls. Associations between animal and plant protein intakes, and anthropometry and serum biomarkers were examined with General linear Model multivariate analysis.
Results
Average total protein intake exceeded the recommendations of World Health Organization and European Food Safety Authority. Mean total protein intake was 96 g/d (59% derived from animal protein). Total, animal and plant protein intakes (g/d) were significantly lower in females than in males and total and plant protein intakes were lower in younger participants (12.5-14.9 y). Protein intake was significantly lower in underweight subjects and higher in obese ones; the direction of the relationship was reversed after adjustments for body weight (g/(kg.d)). The inverse association of plant protein intakes was stronger with BMI z-score and body fat percentage (BF%) compared to animal protein intakes. Additionally, BMI and BF% were positively associated with energy percentage of animal protein.
Conclusions
This sample of European adolescents appeared to have adequate total protein intake. Our findings suggest that plant protein intakes may play a role in preventing obesity among European adolescents. Further longitudinal studies are needed to investigate the potential beneficial effects observed in this study in the prevention of obesity and related chronic diseases.
doi:10.1186/1475-2891-14-10
PMCID: PMC4334414  PMID: 25609179
Protein intake; Adolescence; Body composition; Biomarkers; HELENA study
7.  Assessment of the feasibility of high-concentration capsaicin patches in the pain unit of a tertiary hospital for a population of mixed refractory peripheral neuropathic pain syndromes in Non-diabetic patients 
BMC Anesthesiology  2014;14:120.
Background
High-concentration-capsaicin-patches (Qutenza®) have been put on the market as a treatment for peripheral neuropathic pain. A minimum infrastructure and a determinate skill set for its application are required. Our aim was to assess the feasibility of treatment with high-concentration-capsaicin-patches in clinical practice in a variety of refractory peripheral neuropathic pain syndromes in non-diabetic patients.
Methods
Observational, prospective, single-center study of patients attended to in the Pain Unit of a tertiary hospital, ≥18 year-old non-responders to multimodal analgesia of both genders. The feasibility for the application of capsaicin patch in clinical practice was evaluated by means of the number of patients controlled per day when this one was applied and by means of the times used for patch application.
Results
Between October 2010 and September 2011, 20 consecutive non-diabetic patients (7 males, 13 females) with different diagnoses of refractory peripheral neuropathic pain syndromes, with a median (range) age of 60 (33–88) years-old were treated with a single patch application. The median (range) number of patients monitored per day was not modified when the capsaicin patch was applied [27 (26–29)] in comparison with it was not applied [28 (26–30)]. The median (range) total time to determine and mark the painful area was 9 (6–15) minutes and of patch application was 60 (58–65) minutes. No important adverse reactions were observed.
Conclusion
High-concentration-capsaicin-patch treatment was feasible in our unit for the treatment of a population with refractory peripheral neuropathic pain. The routine of our unit was not affected by its use.
doi:10.1186/1471-2253-14-120
PMCID: PMC4289547  PMID: 25580085
Peripheral neuropathic pain; Capsaicin patch; Clinical practice
8.  Energy Balance Related Behaviour: Personal, Home- and Friend-Related Factors among Schoolchildren in Europe Studied in the ENERGY-Project 
PLoS ONE  2014;9(11):e111775.
Objective
To design interventions that target energy balance-related behaviours, knowledge of primary schoolchildren's perceptions regarding soft drink intake, fruit juice intake, breakfast consumption, TV viewing and physical activity (PA) is essential. The current study describes personal beliefs and attitudes, home- and friend-related variables regarding these behaviours across Europe.
Design
Cross-sectional study in which personal, family and friend -related variables were assessed by validated questionnaires, and dichotomized as favourable versus unfavourable answers. Logistic regression analyses were conducted to estimate proportions of children giving unfavourable answers and test between-country differences.
Setting
A survey in eight European countries.
Subjects
A total of 7903 10–12 year old primary schoolchildren.
Results
A majority of the children reported unfavourable attitudes, preferences and subjective norms regarding soft drink, fruit juice intake and TV viewing accompanied with high availability and accessibility at home. Few children reported unfavourable attitudes and preferences regarding breakfast consumption and PA. Many children reported unfavourable health beliefs regarding breakfast consumption and TV viewing. Substantial differences between countries were observed, especially for variables regarding soft drink intake, breakfast consumption and TV viewing.
Conclusion
The surveyed children demonstrated favourable attitudes to some healthy behaviours (PA, breakfast intake) as well as to some unhealthy behaviours (soft drink consumption, TV viewing). Additionally, many children across Europe have personal beliefs and are exposed to social environments that are not supportive to engagement in healthy behaviours. Moreover, the large differences in personal, family and friend-related variables across Europe argue for implementing different strategies in the different European countries.
doi:10.1371/journal.pone.0111775
PMCID: PMC4221133  PMID: 25372490
9.  Agreement between parent and child report on parental practices regarding dietary, physical activity and sedentary behaviours: the ENERGY cross-sectional survey 
BMC Public Health  2014;14:918.
Background
Parents and their parenting practices play an important role in shaping their children’s environment and energy-balance related behaviours (EBRBs). Measurement of parenting practices can be parent- or child-informed, however not much is known about agreement between parent and child perspectives. This study aimed to assess agreement between parent and child reports on parental practices regarding EBRBs across different countries in Europe and to identify correlates of agreement.
Methods
Within the ENERGY-project, a cross-sectional survey was conducted among 10–12 year old children and their parents in eight European countries. Both children and parents filled in a questionnaire on 14 parental practices regarding five different EBRBs (i.e. soft drink, fruit juice and breakfast consumption, sports activity and watching TV) and socio-demographic characteristics. Children’s anthropometric measurements were taken at school. We calculated percentages of agreement between children and their parents and weighted kappa statistics (for ordinal variables) per practice and country and assessed factors associated with agreement using multilevel linear regression.
Results
Reports of 6425 children and their parents were available for analysis. Overall mean agreement between parent and child reports was 43% and varied little among countries. The lowest agreement was found for questions assessing joint parent–child activities, such as sports (27%; Kappa (κ) = 0.14) or watching TV (30%;κ = 0.17), and for parental allowance of the child to have soft drinks (32%;κ = 0.24) or fruit juices (32%;κ = 0.19), or to watch TV (27%;κ = 0.17). Having breakfast products available at home or having a TV in the child’s bedroom were the only practices with moderate to good agreement (>60%;κ = 0.06 and 0.77, respectively). In general, agreement was lower for boys, younger children, younger parents, parents with less than 14 years of education, single parents, parents with a higher self-reported body mass index and parents who perceived their child to be underweight.
Conclusions
Parents and children perceive parental practices regarding dietary, physical activity and sedentary behaviours differently in all parts of Europe, with considerable variation across specific practices and countries. Therefore, future studies should assess both, parents and children’s view on parental practices.
Electronic supplementary material
The online version of this article (doi:10.1186/1471-2458-14-918) contains supplementary material, which is available to authorized users.
doi:10.1186/1471-2458-14-918
PMCID: PMC4169834  PMID: 25190541
Children; Parent; Parenting; Questionnaire; Inter-observer agreement; Inter-observer variability; Health behaviour; Overweight
10.  Parents and friends both matter: simultaneous and interactive influences of parents and friends on European schoolchildren’s energy balance-related behaviours – the ENERGY cross-sectional study 
Background
The family, and parents in particular, are considered the most important influencers regarding children’s energy-balance related behaviours (EBRBs). When children become older and gain more behavioural autonomy regarding different behaviours, the parental influences may become less important and peer influences may gain importance. Therefore the current study aims to investigate simultaneous and interactive associations of family rules, parent and friend norms and modelling with soft drink intake, TV viewing, daily breakfast consumption and sport participation among schoolchildren across Europe.
Methods
A school-based cross-sectional survey in eight countries across Europe among 10–12 year old schoolchildren. Child questionnaires were used to assess EBRBs (soft drink intake, TV viewing, breakfast consumption, sport participation), and potential determinants of these behaviours as perceived by the child, including family rules, parental and friend norms and modelling. Linear and logistic regression analyses (n = 7811) were applied to study the association of parental (norms, modelling and rules) and friend influences (norm and modelling) with the EBRBs. In addition, potential moderating effects of parental influences on the associations of friend influences with the EBRBs were studied by including interaction terms.
Results
Children reported more unfavourable friend norms and modelling regarding soft drink intake and TV viewing, while they reported more favourable friend and parental norms and modelling for breakfast consumption and physical activity. Perceived friend and parental norms and modelling were significantly positively associated with soft drink intake, breakfast consumption, physical activity (only modelling) and TV time. Across the different behaviours, ten significant interactions between parental and friend influencing variables were found and suggested a weaker association of friend norms and modelling when rules were in place.
Conclusion
Parental and friends norm and modelling are associated with schoolchildren’s energy balance-related behaviours. Having family rules or showing favourable parental modelling and norms seems to reduce the potential unfavourable associations of friends’ norms and modelling with the EBRBs.
doi:10.1186/1479-5868-11-82
PMCID: PMC4098693  PMID: 25001090
Parents; Friends; Social norm; Modelling; Rules; Soft drink; TV viewing; Physical activity
11.  Innovative Pressure Sensor Platform and Its Integration with an End-User Application 
Sensors (Basel, Switzerland)  2014;14(6):10273-10291.
This paper describes the fully integration of an innovative and low-cost pressure sensor sheet based on a bendable and printed electronics technology. All integration stages are covered, from most low-level functional system, like physical analog sensor data acquisition, followed by embedded data processing, to end user interactive visual application. Data acquisition embedded software and hardware was developed using a Rapid Control Prototyping (RCP). Finally, after first electronic prototype successful testing, a Taylor-made electronics was developed, reducing electronics volume to 3.5 cm × 6 cm × 2 cm with a maximum power consumption of 765 mW for both electronics and pressure sensor sheet.
doi:10.3390/s140610273
PMCID: PMC4118367  PMID: 24922455
systems integration; pressure sensor; bendable sensor; Rapid Control Prototyping; embedded software; end-user application
12.  Potential biases in the classification, analysis and interpretations in cross-sectional study: commentaries – surrounding the article "resting heart rate: its correlations and potential for screening metabolic dysfunctions in adolescents" 
BMC Pediatrics  2014;14:117.
Background
Resting heart rate reflects sympathetic nerve activity. A significant association between resting heart rate (HR) and all causes of cardiovascular mortality has been reported by some epidemiologic studies. Despite suggestive evidence, resting heart rate (RHR) has not been formally explored as a prognostic factor and potential therapeutic outcome and, therefore, is not generally accepted in adolescents.
Discussion
The core of the debate is the methodological aspects used in "Resting heart rate: its correlations and potential for screening metabolic dysfunctions in adolescents"; the points are: cutoff used for cluster RHR, two different statistical models used to analyze the same set of variables, one for continuous data, and another for categorical data; interpretation of p-value < 0.05, sampling process involving two random stages, analysis of design effect and the parameters of screening tests.
Summary
Aspects that must be taken into account for evaluation of a screening test to measure the potential for discrimination for a common variable (population with outcome vs. no outcome population), the main indicators are: sensitivity, specificity, accuracy, positive predictive value and negative predictive value. The measures of argumentation equality (CI) or difference (p-valor) are important to validate these indicators but do not indicate quality of screening.
doi:10.1186/1471-2431-14-117
PMCID: PMC4012522  PMID: 24885992
Resting heart rate; Screening test; P-value; High glucose; High triglycerides
13.  Genetic and Molecular Insights Into the Role of PROX1 in Glucose Metabolism 
Diabetes  2013;62(5):1738-1745.
Genome-wide association studies have shown that the rs340874 single nucleotide polymorphism (SNP) in PROX1 is a genetic susceptibility factor for type 2 diabetes. We conducted genetic and molecular studies to better understand the role of PROX1 in type 2 diabetes. We assessed the impact of the whole common genetic variability of PROX1 (80 SNPs) on type 2 diabetes–related biochemical traits in the HELENA (Healthy Lifestyle in Europe by Nutrition in Adolescence) study (n = 1,155). Three SNPs (rs340838, rs340837, and rs340836) were significantly associated with fasting plasma insulin levels (P ≤ 0.00295). We evaluated the impact of nine PROX1 SNPs (the three insulin-associated SNPs plus six SNPs in strong linkage disequilibrium) on luciferase reporter gene expression. The insulin-lowering alleles of rs340874, rs340873, and rs340835 were associated with lower luciferase activity in MIN6 and HepG2 cells (except for rs340874, which was in HepG2 cells only). Electrophoretic mobility shift assays indicated that specific nuclear protein bindings occur at the three SNPs in HepG2 cells, with allele-binding differences for rs340874. We also showed that the knockdown of Prox1 expression by small interfering RNAs in INS-1E cells resulted in a 1.7-fold reduction in glucose-stimulated insulin secretion. All together, we propose that reduced expression of PROX1 by cis-regulatory variants results in altered β-cell insulin secretion and thereby confers susceptibility to type 2 diabetes.
doi:10.2337/db12-0864
PMCID: PMC3636631  PMID: 23274905
14.  Telomere Length as a Biomarker for Adiposity Changes after a Multidisciplinary Intervention in Overweight/Obese Adolescents: The EVASYON Study 
PLoS ONE  2014;9(2):e89828.
Context
Telomeres are biomarkers of biological aging. Shorter telomeres have been associated with increased adiposity in adults. However, this relationship remains unclear in children and adolescents.
Objective
To evaluate the association between telomere length (TL) and adiposity markers in overweight/obese adolescents after an intensive program. We hypothesize that greater TL at baseline would predict a better response to a weight loss treatment.
Design, Setting, Patients and Intervention
The EVASYON is a multidisciplinary treatment program for adolescents with overweight and obesity that is aimed at applying the intervention to all possibly involved areas of the individual, such as dietary habits, physical activity and cognitive and psychological profiles. Seventy-four participants (36 males, 38 females, 12–16 yr) were enrolled in the intervention program: 2 months of an energy-restricted diet and a follow-up period (6 months).
Main Outcome
TL was measured by quantitative real-time polymerase chain reaction at baseline and after 2 months; meanwhile, anthropometric variables were also assessed after 6 months of follow-up.
Results
TL lengthened in participants during the intensive period (+1.9±1.0, p<0.001) being greater in overweight/obese adolescents with the shortest telomeres at baseline (r = −0.962, p<0.001). Multivariable linear regression analysis showed that higher baseline TL significantly predicted a higher decrease in body weight (B = −1.53, p = 0.005; B = −2.25, p = 0.047) and in standard deviation score for body mass index (BMI-SDS) (B = −0.22, p = 0.010; B = −0.47, p = 0.005) after the intensive and extensive period treatment respectively, in boys.
Conclusion
Our study shows that a weight loss intervention is accompanied by a significant increase in TL in overweight/obese adolescents. Moreover, we suggest that initial longer TL could be a potential predictor for a better weight loss response.
doi:10.1371/journal.pone.0089828
PMCID: PMC3933678  PMID: 24587065
15.  Early Life Course Risk Factors for Childhood Obesity: The IDEFICS Case-Control Study 
PLoS ONE  2014;9(2):e86914.
Background
The early life course is assumed to be a critical phase for childhood obesity; however the significance of single factors and their interplay is not well studied in childhood populations.
Objectives
The investigation of pre-, peri- and postpartum risk factors on the risk of obesity at age 2 to 9.
Methods
A case-control study with 1,024 1∶1-matched case-control pairs was nested in the baseline survey (09/2007–05/2008) of the IDEFICS study, a population-based intervention study on childhood obesity carried out in 8 European countries in pre- and primary school settings. Conditional logistic regression was used for identification of risk factors.
Results
For many of the investigated risk factors, we found a raw effect in our study. In multivariate models, we could establish an effect for gestational weight gain (adjusted OR = 1.02; 95%CI 1.00–1.04), smoking during pregnancy (adjusted OR = 1.48; 95%CI 1.08–2.01), Caesarian section (adjusted OR = 1.38; 95%CI 1.10–1.74), and breastfeeding 4 to 11 months (adjusted OR = 0.77; 95%CI 0.62–0.96). Birth weight was related to lean mass rather than to fat mass, the effect of smoking was found only in boys, but not in girls. After additional adjustment for parental BMI and parental educational status, only gestational weight gain remained statistically significant. Both, maternal as well as paternal BMI were the strongest risk factors in our study, and they confounded several of the investigated associations.
Conclusions
Key risk factors of childhood obesity in our study are parental BMI and gestational weight gain; consequently prevention approaches should target not only children but also adults. The monitoring of gestational weight seems to be of particular importance for early prevention of childhood obesity.
doi:10.1371/journal.pone.0086914
PMCID: PMC3923715  PMID: 24551043
16.  Myeloperoxidase Is an Early Biomarker of Inflammation and Cardiovascular Risk in Prepubertal Obese Children 
Diabetes Care  2012;35(11):2373-2376.
OBJECTIVE
Obesity is associated with a state of chronic low-grade inflammation. Myeloperoxidase (MPO) plays an important role in the initiation and progression of acute and chronic inflammatory diseases, such as cardiovascular disease (CVD). The objectives of the current study were to evaluate plasma MPO levels in prepubertal obese children and to determine whether MPO could be an early biomarker of inflammation and CVD risk.
RESEARCH DESIGN AND METHODS
In a prospective multicenter case-control study paired by age and sex of 446 Caucasian prepubertal children ages 6–12 years, 223 normal-weight and 223 obese children were recruited. Blood pressure, waist circumference, weight, and height were measured. In addition to MPO, glucose, insulin, metabolic lipid parameters, oxidized low-density lipoproteins, adiponectin, leptin, resistin, C-reactive protein (CRP), interleukin 6, tumor necrosis factor α, matrix metalloproteinase-9 (MMP-9), and plasminogen activator inhibitor 1 were determined.
RESULTS
We found that MPO was elevated in prepubertal obese children and that this enzyme was associated with such proinflammatory and cardiovascular risk biomarkers as CRP, MMP-9, and resistin. Insulin resistance calculated by the homeostatic assessment model was the best predictor of MPO.
CONCLUSIONS
MPO is an early biomarker of inflammation associated with CVD risk in obese children at the prepubertal age.
doi:10.2337/dc12-0614
PMCID: PMC3476926  PMID: 22912422
17.  Can Ethnic Background Differences in Children’s Body Composition Be Explained by Differences in Energy Balance-Related Behaviors? A Mediation Analysis within the Energy-Project 
PLoS ONE  2013;8(8):e71848.
Background
In affluent countries, children from non-native ethnicity have in general less favourable body composition indicators and energy balance-related behaviors (EBRBs) than children from native ethnicity. However, differences between countries have been reported.
Methodology/Principal Findings
A school-based survey among 10–12 years old children was conducted in seven European countries with a standardized protocol. Weight, height and waist circumference were measured; engagement in EBRBs was self-reported. For those countries with significant ethnic differences in body composition (Greece and the Netherlands), multilevel mediation analyses were conducted, to test the mediating effect of the EBRBs in the association between ethnic background and body composition indicators. Analyses were adjusted for gender and age, and for parental education in a later step. Partial mediation was found for sugared drinks intake and sleep duration in the Greek sample, and breakfast in the Dutch sample. A suppression effect was found for engagement in sports activites in the Greek sample.
Conclusions/Significance
Ethnic differences in children’s body composition were partially mediated by differences in breakfast skipping in the Netherlands and sugared drinks intake, sports participation and sleep duration in Greece.
doi:10.1371/journal.pone.0071848
PMCID: PMC3744449  PMID: 23967254
18.  Understanding the Links among neuromedin U Gene, beta2-adrenoceptor Gene and Bone Health: An Observational Study in European Children 
PLoS ONE  2013;8(8):e70632.
Neuromedin U, encoded by the NMU gene, is a hypothalamic neuropeptide that regulates both energy metabolism and bone mass. The beta-2 adrenergic receptor, encoded by the ADRB2 gene, mediates several effects of catecholamine hormones and neurotransmitters in bone. We investigated whether NMU single nucleotide polymorphisms (SNPs) and haplotypes, as well as functional ADRB2 SNPs, are associated with bone stiffness in children from the IDEFICS cohort, also evaluating whether NMU and ADRB2 interact to affect this trait. A sample of 2,274 subjects (52.5% boys, age 6.2±1.8 years) from eight European countries, having data on calcaneus bone stiffness index (SI, mean of both feet) and genotyping (NMU gene: rs6827359, rs12500837, rs9999653; ADRB2 gene: rs1042713, rs1042714), was studied. After false discovery rate adjustment, SI was significantly associated with all NMU SNPs. rs6827359 CC homozygotes showed the strongest association (recessive model, Δ = −1.8, p = 0.006). Among the five retrieved haplotypes with frequencies higher than 1% (range 2.0–43.9%), the CCT haplotype (frequency = 39.7%) was associated with lower SI values (dominant model, Δ = −1.0, p = 0.04) as compared to the most prevalent haplotype. A non-significant decrease in SI was observed in in ADRB2 rs1042713 GG homozygotes, while subjects carrying SI-lowering genotypes at both SNPs (frequency = 8.4%) showed much lower SI than non-carriers (Δ = −3.9, p<0.0001; p for interaction = 0.025). The association was more evident in preschool girls, in whom SI showed a curvilinear trend across ages. In subgroup analyses, rs9999653 CC NMU or both GG ADRB2 genotypes were associated with either lower serum calcium or β-CrossLaps levels (p = 0.01). This study in European children shows, for the first time in humans, a role for NMU gene through interaction with ADRB2 gene in bone strength regulation, more evident in preschool girls.
doi:10.1371/journal.pone.0070632
PMCID: PMC3731254  PMID: 23936460
19.  Physical activity and clustered cardiovascular disease risk factors in young children: a cross-sectional study (the IDEFICS study) 
BMC Medicine  2013;11:172.
Background
The relevance of physical activity (PA) for combating cardiovascular disease (CVD) risk in children has been highlighted, but to date there has been no large-scale study analyzing that association in children aged ≤9 years of age. This study sought to evaluate the associations between objectively-measured PA and clustered CVD risk factors in a large sample of European children, and to provide evidence for gender-specific recommendations of PA.
Methods
Cross-sectional data from a longitudinal study in 16,224 children aged 2 to 9 were collected. Of these, 3,120 (1,016 between 2 to 6 years, 2,104 between 6 to 9 years) had sufficient data for inclusion in the current analyses. Two different age-specific and gender-specific clustered CVD risk scores associated with PA were determined. First, a CVD risk factor (CRF) continuous score was computed using the following variables: systolic blood pressure (SBP), total triglycerides (TG), total cholesterol (TC)/high-density lipoprotein cholesterol (HDL-c) ratio, homeostasis model assessment of insulin resistance (HOMA-IR), and sum of two skinfolds (score CRFs). Secondly, another CVD risk score was obtained for older children containing the score CRFs + the cardiorespiratory fitness variable (termed score CRFs + fit). Data used in the current analysis were derived from the IDEFICS (‘Identification and prevention of Dietary- and lifestyle-induced health EFfects In Children and infantS’) study.
Results
In boys <6 years, the odds ratios (OR) for CVD risk were elevated in the least active quintile of PA (OR: 2.58) compared with the most active quintile as well as the second quintile for vigorous PA (OR: 2.91). Compared with the most active quintile, older children in the first, second and third quintiles had OR for CVD risk score CRFs + fit ranging from OR 2.69 to 5.40 in boys, and from OR 2.85 to 7.05 in girls.
Conclusions
PA is important to protect against clustering of CVD risk factors in young children, being more consistent in those older than 6 years. Healthcare professionals should recommend around 60 and 85 min/day of moderate-to-vigorous PA, including 20 min/day of vigorous PA.
Please see related commentary: http://www.biomedcentral.com/1741-7015/11/173.
doi:10.1186/1741-7015-11-172
PMCID: PMC3728104  PMID: 23899208
Accelerometers; Cardiovascular disease risk factors; Physical activity; Younger children
20.  Association of objectively measured physical activity with body components in European adolescents 
BMC Public Health  2013;13:667.
Background
Physical activity (PA) is suggested to contribute to fat loss not only through increasing energy expenditure “per se” but also increasing muscle mass; therefore, it would be interesting to better understand the specific associations of PA with the different body’s components such as fat mass and muscle mass. The aim of the present study was to examine the association between objectively measured PA and indices of fat mass and muscle components independently of each other giving, at the same time, gender-specific information in a wide cohort of European adolescents.
Methods
A cross-sectional study in a school setting was conducted in 2200 (1016 males) adolescents (14.7 ±1.2 years). Weight, height, skinfold thickness, bioimpedance and PA (accelerometry) were measured. Indices of fat mass (body mass index, % fat mass, sum of skinfolds) and muscular component (assessed as fat-free mass) were calculated. Multiple regression analyses were performed adjusting for several confounders including fat-free mass and fat mass when possible.
Results
Vigorous PA was positively associated with height (p < 0.05) in males, whilst, vigorous PA, moderate-vigorous PA and average PA were negatively associated with all the indices of fat mass (all p < 0.01) in both genders, except for average PA in relation with body mass index in females. Regarding muscular components, vigorous PA showed positive associations with fat-free mass and muscle mass (all p < 0.05) in both genders. Average PA was positively associated with fat-free mass (both p < 0.05) in males and females.
Conclusion
The present study suggests that PA, especially vigorous PA, is negatively associated with indices of fat mass and positively associated with markers of muscle mass, after adjusting for several confounders (including indices of fat mass and muscle mass when possible). Future studies should focus not only on the classical relationship between PA and fat mass, but also on PA and muscular components, analyzing the independent role of both with the different PA intensities.
doi:10.1186/1471-2458-13-667
PMCID: PMC3723445  PMID: 23866681
Physical activity; Fat mass; Muscular components; Adolescence
21.  New loci associated with birth weight identify genetic links between intrauterine growth and adult height and metabolism 
Horikoshi, Momoko | Yaghootkar, Hanieh | Mook-Kanamori, Dennis O. | Sovio, Ulla | Taal, H. Rob | Hennig, Branwen J. | Bradfield, Jonathan P. | St. Pourcain, Beate | Evans, David M. | Charoen, Pimphen | Kaakinen, Marika | Cousminer, Diana L. | Lehtimäki, Terho | Kreiner-Møller, Eskil | Warrington, Nicole M. | Bustamante, Mariona | Feenstra, Bjarke | Berry, Diane J. | Thiering, Elisabeth | Pfab, Thiemo | Barton, Sheila J. | Shields, Beverley M. | Kerkhof, Marjan | van Leeuwen, Elisabeth M. | Fulford, Anthony J. | Kutalik, Zoltán | Zhao, Jing Hua | den Hoed, Marcel | Mahajan, Anubha | Lindi, Virpi | Goh, Liang-Kee | Hottenga, Jouke-Jan | Wu, Ying | Raitakari, Olli T. | Harder, Marie N. | Meirhaeghe, Aline | Ntalla, Ioanna | Salem, Rany M. | Jameson, Karen A. | Zhou, Kaixin | Monies, Dorota M. | Lagou, Vasiliki | Kirin, Mirna | Heikkinen, Jani | Adair, Linda S. | Alkuraya, Fowzan S. | Al-Odaib, Ali | Amouyel, Philippe | Andersson, Ehm Astrid | Bennett, Amanda J. | Blakemore, Alexandra I.F. | Buxton, Jessica L. | Dallongeville, Jean | Das, Shikta | de Geus, Eco J. C. | Estivill, Xavier | Flexeder, Claudia | Froguel, Philippe | Geller, Frank | Godfrey, Keith M. | Gottrand, Frédéric | Groves, Christopher J. | Hansen, Torben | Hirschhorn, Joel N. | Hofman, Albert | Hollegaard, Mads V. | Hougaard, David M. | Hyppönen, Elina | Inskip, Hazel M. | Isaacs, Aaron | Jørgensen, Torben | Kanaka-Gantenbein, Christina | Kemp, John P. | Kiess, Wieland | Kilpeläinen, Tuomas O. | Klopp, Norman | Knight, Bridget A. | Kuzawa, Christopher W. | McMahon, George | Newnham, John P. | Niinikoski, Harri | Oostra, Ben A. | Pedersen, Louise | Postma, Dirkje S. | Ring, Susan M. | Rivadeneira, Fernando | Robertson, Neil R. | Sebert, Sylvain | Simell, Olli | Slowinski, Torsten | Tiesler, Carla M.T. | Tönjes, Anke | Vaag, Allan | Viikari, Jorma S. | Vink, Jacqueline M. | Vissing, Nadja Hawwa | Wareham, Nicholas J. | Willemsen, Gonneke | Witte, Daniel R. | Zhang, Haitao | Zhao, Jianhua | Wilson, James F. | Stumvoll, Michael | Prentice, Andrew M. | Meyer, Brian F. | Pearson, Ewan R. | Boreham, Colin A.G. | Cooper, Cyrus | Gillman, Matthew W. | Dedoussis, George V. | Moreno, Luis A | Pedersen, Oluf | Saarinen, Maiju | Mohlke, Karen L. | Boomsma, Dorret I. | Saw, Seang-Mei | Lakka, Timo A. | Körner, Antje | Loos, Ruth J.F. | Ong, Ken K. | Vollenweider, Peter | van Duijn, Cornelia M. | Koppelman, Gerard H. | Hattersley, Andrew T. | Holloway, John W. | Hocher, Berthold | Heinrich, Joachim | Power, Chris | Melbye, Mads | Guxens, Mònica | Pennell, Craig E. | Bønnelykke, Klaus | Bisgaard, Hans | Eriksson, Johan G. | Widén, Elisabeth | Hakonarson, Hakon | Uitterlinden, André G. | Pouta, Anneli | Lawlor, Debbie A. | Smith, George Davey | Frayling, Timothy M. | McCarthy, Mark I. | Grant, Struan F.A. | Jaddoe, Vincent W.V. | Jarvelin, Marjo-Riitta | Timpson, Nicholas J. | Prokopenko, Inga | Freathy, Rachel M.
Nature genetics  2012;45(1):76-82.
Birth weight within the normal range is associated with a variety of adult-onset diseases, but the mechanisms behind these associations are poorly understood1. Previous genome-wide association studies identified a variant in the ADCY5 gene associated both with birth weight and type 2 diabetes, and a second variant, near CCNL1, with no obvious link to adult traits2. In an expanded genome-wide association meta-analysis and follow-up study (up to 69,308 individuals of European descent from 43 studies), we have now extended the number of genome-wide significant loci to seven, accounting for a similar proportion of variance to maternal smoking. Five of the loci are known to be associated with other phenotypes: ADCY5 and CDKAL1 with type 2 diabetes; ADRB1 with adult blood pressure; and HMGA2 and LCORL with adult height. Our findings highlight genetic links between fetal growth and postnatal growth and metabolism.
doi:10.1038/ng.2477
PMCID: PMC3605762  PMID: 23202124
22.  Impact of Social Marketing in the Prevention of Childhood Obesity123 
Advances in Nutrition  2012;3(4):611S-615S.
Obesity, mainly childhood obesity, is a worldwide concern. Childhood obesity continues to adulthood, and it is associated with multiple noncommunicable diseases. One important aspect in the fight against obesity is prevention, the earlier, the better. Social marketing is a novel concept being increasingly used as an approach to address social problems and more and more included in the community-based interventions aiming to change unhealthy behaviors. Although there is limited evidence of its effectiveness, it seems that when conscientiously applied, social marketing principles may be useful to change behaviors and thus better health outcomes.
doi:10.3945/an.112.001958
PMCID: PMC3649734  PMID: 22798001
23.  Parental education associations with children’s body composition: mediation effects of energy balance-related behaviors within the ENERGY-project 
Background
It is well known that the prevalence of overweight and obesity is considerably higher among youth from lower socio-economic families, but there is little information about the role of some energy balance-related behaviors in the association between socio-economic status and childhood overweight and obesity. The objective of this paper was to assess the possible mediation role of energy balance-related behaviors in the association between parental education and children’s body composition.
Methods
Data were obtained from the cross sectional study of the “EuropeaN Energy balance Research to prevent excessive weight Gain among Youth” (ENERGY) project. 2121 boys and 2516 girls aged 10 to 12 from Belgium, Greece, Hungary, the Netherlands, Norway, Slovenia and Spain were included in the analyses. Data were obtained via questionnaires assessing obesity related dietary, physical activity and sedentary behaviors and basic anthropometric objectively measured indicators (weight, height, waist circumference). The possible mediating effect of sugared drinks intake, breakfast consumption, active transportation to school, sports participation, TV viewing, computer use and sleep duration in the association between parental education and children’s body composition was explored via MacKinnon’s product-of-coefficients test in single and multiple mediation models. Two different body composition indicators were included in the models, namely Body Mass Index and waist circumference.
Results
The association between parental education and children’s body composition was partially mediated by breakfast consumption, sports participation, TV viewing and computer use. Additionally, a suppression effect was found for sugared drinks intake. No mediation effect was found for active transportation and sleep duration. The significant mediators explained a higher proportion of the association between parental education and waist circumference compared to the association between parental education and BMI.
Conclusions
Tailored overweight and obesity prevention strategies in low SES preadolescent populations should incorporate specific messages focusing on the importance of encouraging daily breakfast consumption, increasing sports participation and decreasing TV viewing and computer use. However, longitudinal research to support these findings is needed.
doi:10.1186/1479-5868-10-80
PMCID: PMC3695820  PMID: 23800170
Mediation analysis; Body composition; Parental education; Childhood obesity; Energy balance-related behaviors
24.  High‐sensitivity C‐reactive Protein is a Predictive Factor of Adiposity in Children: Results of the Identification and prevention of Dietary‐ and lifestyle‐induced health Effects in Children and InfantS (IDEFICS) Study 
Background
Whereas cross‐sectional studies have shown that obesity is associated with increased C‐reactive protein (CRP) levels in children, little is known about the impact of low‐grade inflammation on body mass changes during growth.
Methods and Results
We assessed cross‐sectionally and longitudinally the association of high‐sensitivity (hs)‐CRP levels with overweight/obesity and related cardiometabolic risk factors in the Identification and prevention of Dietary‐ and lifestyle‐induced health Effects in Children and InfantS (IDEFICS) cohort. 16 224 children from 8 European countries (2 to 9 years) were recruited during the baseline survey (T0). After the exclusion of 7187 children because of missing hs‐CRP measurements and 2421 because of drug use during the previous week, the analysis was performed on 6616 children (Boys=3347; Girls=3269; age=6.3±1.7 years). Of them, 4110 were reexamined 2 years later (T1). Anthropometric variables, blood pressure, hs‐CRP, blood lipids, glucose and insulin were measured. The population at T0 was divided into 3 categories, according to the baseline hs‐CRP levels. Higher hs‐CRP levels were associated with significantly higher prevalence of overweight/obesity, body mass index (BMI) z‐score and central adiposity indices (P values all <0.0001), and with higher blood pressure and lower HDL‐cholesterol levels. Over the 2‐year follow‐up, higher baseline hs‐CRP levels were associated with a significant increase in BMI z‐score (P<0.001) and significantly higher risk of incident overweight/obesity.
Conclusions
Higher hs‐CRP levels are associated to higher body mass and overweight/obesity risk in a large population of European children. Children with higher baseline levels of hs‐CRP had a greater increase in BMI z‐score and central adiposity over time and were at higher risk of developing overweight/obesity during growth.
doi:10.1161/JAHA.113.000101
PMCID: PMC3698769  PMID: 23744403
C‐reactive protein; inflammation; pediatric obesity; population
25.  Associations between eating meals, watching TV while eating meals and weight status among children, ages 10–12 years in eight European countries: the ENERGY cross-sectional study 
Background
To assess the association of eating meals, and never watching TV while eating meals, with weight status among children, ages 10–12 years across Europe.
Methods
7915 children (mean age: 11.5 years) in eight European countries (Belgium, Greece, Hungary, the Netherlands, Norway, Slovenia, Spain and Switzerland) completed a questionnaire at school. Data on meals eaten the day before questionnaire administration and the frequency of eating meals while watching TV were collected. Height and weight of the children were objectively assessed. Multinomial and binary regression analyses were conducted to test associations of eating meals (adjusted for gender and ethnicity) and never watching TV while eating meals (adjusted for gender, ethnicity and total TV time) with overweight/obesity, and to test for country- and socio-demographic differences.
Results
The proportions of children reporting eating breakfast, lunch and dinner were 85%, 96%, and 93% respectively, and 55%, 46% and 32% reported to never watch TV at breakfast, lunch and dinner respectively. The children who ate breakfast (OR = 0.6 (95% CI 0.5-0.7)) and dinner (OR = 0.4 (95% CI 0.3-0.5)), had lower odds of being overweight compared to those who did not. The children who never watched TV at lunch (OR = 0.7 (95% CI 0.7-0.8)) and dinner (OR = 0.8 (95% CI 0.7-0.9)) had lower odds of being overweight compared to those who watched TV at the respective meals.
Conclusions
The odds of being overweight was lower for children who ate breakfast and dinner compared to those who did not eat the respective meals. The odds of being overweight was lower for children who reported to never watch TV at lunch and dinner compared to those who did. A focus towards meal frequency and watching TV during meals in longitudinal and interventions studies in prevention of overweight and obesity, may contribute to a better understanding of causality.
doi:10.1186/1479-5868-10-58
PMCID: PMC3663732  PMID: 23675988
Children; Weight status; Meals; Breakfast; TV viewing; Never TV at meals

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