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1.  Cycling and bone health: a systematic review 
BMC Medicine  2012;10:168.
Background
Cycling is considered to be a highly beneficial sport for significantly enhancing cardiovascular fitness in individuals, yet studies show little or no corresponding improvements in bone mass.
Methods
A scientific literature search on studies discussing bone mass and bone metabolism in cyclists was performed to collect all relevant published material up to April 2012. Descriptive, cross-sectional, longitudinal and interventional studies were all reviewed. Inclusion criteria were met by 31 studies.
Results
Heterogeneous studies in terms of gender, age, data source, group of comparison, cycling level or modality practiced among others factors showed minor but important differences in results. Despite some controversial results, it has been observed that adult road cyclists participating in regular training have low bone mineral density in key regions (for example, lumbar spine). Conversely, other types of cycling (such as mountain biking), or combination with other sports could reduce this unsafe effect. These results cannot yet be explained by differences in dietary patterns or endocrine factors.
Conclusions
From our comprehensive survey of the current available literature it can be concluded that road cycling does not appear to confer any significant osteogenic benefit. The cause of this may be related to spending long hours in a weight-supported position on the bike in combination with the necessary enforced recovery time that involves a large amount of time sitting or lying supine, especially at the competitive level.
doi:10.1186/1741-7015-10-168
PMCID: PMC3554602  PMID: 23256921
cyclists; osteopenia; osteoporosis; sport; training
2.  Prevalence of negative life events and chronic adversities in European pre- and primary-school children: results from the IDEFICS study 
Archives of Public Health  2012;70(1):26.
Background
Children are not always recognized as being susceptible to stress, although childhood stressors may originate from multiple events in their everyday surroundings with negative effects on children’s health.
Methods
As there is a lack of large-scale, European prevalence data on childhood adversities, this study presents the prevalence of (1) negative life events and (2) familial and social adversities in 4637 European pre- and primary-school children (4–11 years old), using a parentally-reported questionnaire embedded in the IDEFICS project (‘Identification and prevention of Dietary- and lifestyle-induced health EFfects In Children and infantS’).
Results
The following findings were observed: (1) Certain adversities occur only rarely, while others are very regular (i.e. parental divorce); (2) A large percentage of children is shielded from stressors, while a small group of children is exposed to multiple, accumulating adversities; (3) The prevalence of childhood adversity is influenced by geographical location (e.g. north versus south), age group and sex; (4) Childhood adversities are associated and co-occur, resulting in potential cumulative childhood stress.
Conclusions
This study demonstrated the importance of not only studying traumatic events but also of focusing on the early familial and social environment in childhood stress research and indicated the importance of recording or monitoring childhood adversities.
doi:10.1186/0778-7367-70-26
PMCID: PMC3552702  PMID: 23173879
Child; Life events; Adversity; Prevalence; Stress; Epidemiology
3.  Micro-level economic factors and incentives in Children’s energy balance related behaviours - findings from the ENERGY European cross-section questionnaire survey 
Background
To date, most research on obesogenic environments facing school children has focused on physical and socio-cultural environments. The role of economic factors has been investigated to a much lesser extent. Our objective was to explore the association of micro-level economic factors and incentives with sports activities and intake of soft drinks and fruit juice in 10-12 year-old school children across Europe, and to explore price sensitivity in children’s soft drink consumption and correlates of this price sensitivity.
Methods
Data for the study originate from a cross-sectional survey undertaken in seven European countries (Belgium, Greece, Hungary, Netherlands, Norway, Slovenia and Spain) in 2010 among 10-12 year-old school children and their parents. In total, 7234 child questionnaires and 6002 parent questionnaires were completed. The child questionnaire included questions addressing self-reported weekly intake of soft drinks and fruit juices and time spent on sports activities, perception of parental support for sports activities, use of pocket money for soft drinks and perceived price responsiveness. Parent questionnaires included questions addressing the role of budget and price considerations in decisions regarding children’s sports activities, soft drink consumption, home practices and rules and socio-demographic background variables. Data were analysed using multiple linear regression and discrete-choice (ordered probit) modelling.
Results
Economic factors were found to be associated with children’s sports participation and sugary drink consumption, explaining 27% of the variation in time for sports activities, and 27% and 12% of the variation in the children’s soft drink and juice consumption, respectively. Parents’ financial support was found to be an important correlate (Beta =0.419) of children’s sports activities. Children’s pocket money was a strong correlate (Beta =21.034) of soft drink consumption. The majority of the responding children reported to expect that significantly higher prices of soft drinks would lead them to buy less soft drinks with their own pocket money, but a majority of parents did not expect higher soft drink prices to reduce their children’s soft drink consumption.
Conclusions
We conclude that economic factors, especially parents’ financial support and amount of pocket money, appear to be of importance for children’s sports participation and soft drink consumption, respectively.
doi:10.1186/1479-5868-9-136
PMCID: PMC3514146  PMID: 23171289
Children; Obesogenic environment; Economic incentives; Sports activity; Soft drinks; Price responsiveness
4.  Prospective Analysis of the Association of a Common Variant of FTO (rs9939609) with Adiposity in Children: Results of the IDEFICS Study 
PLoS ONE  2012;7(11):e48876.
Objectives
We investigated cross-sectionally and longitudinally the relationship between FTO rs9939609 and obesity-related characteristics in the European children of the IDEFICS project and the interaction of this variant with a lifestyle intervention.
Population and Methods
A cohort of 16224 children (2–9 years) was recruited into a population-based survey (T0) from eight European countries. A second survey (T1) reassessed the children two years later. A random sample of 4405 children was extracted for genetic studies. 3168 children were re-examined two years later. Half of them underwent a lifestyle intervention program. The FTO rs9939609 was genotyped. Weight, height, waist circumference, triceps and subscapular skinfolds were measured at T0 and T1.
Results
At T0, the risk A allele of rs9939609 was significantly associated with higher values of body mass index (BMI), waist circumference and skinfolds (age, sex, and country-adjusted p-values: all p<0.001) and with a statistically significant increased risk of overweight/obesity.
Over the two year follow-up, no interaction between genotype and intervention was observed. The A allele was associated to a significantly higher increase in all the anthropometric variables examined at T0 independently from the study group (intervention versus control) (p-values: all p<0.002, adjusted for age, sex, country, intervention/control study group, T0 values, and individual time interval between T0 and T1). Over the two-year follow–up, 210 new cases of overweight/obesity occurred. A statistically significant higher incidence of overweight/obesity was associated to the A allele [ORA = 1.95, 95% CI = (1.29; 2.97)].
Conclusions
We confirmed the association between the FTO rs9939609 and body mass and overweight/obesity risk in European children. The main finding of the study is that the A allele carriers present higher increase of body mass and central adiposity over time and higher risk of developing overweight/obesity during growth, independently from intervention measures.
doi:10.1371/journal.pone.0048876
PMCID: PMC3498350  PMID: 23155422
5.  Sedentary behaviours and its association with bone mass in adolescents: the HELENA cross-sectional study 
BMC Public Health  2012;12:971.
Background
We aimed to examine whether time spent on different sedentary behaviours is associated with bone mineral content (BMC) in adolescents, after controlling for relevant confounders such as lean mass and objectively measured physical activity (PA), and if so, whether extra-curricular participation in osteogenic sports could have a role in this association.
Methods
Participants were 359 Spanish adolescents (12.5-17.5 yr, 178 boys,) from the HELENA-CSS (2006–07). Relationships of sedentary behaviours with bone variables were analysed by linear regression. The prevalence of low BMC (at least 1SD below the mean) and time spent on sedentary behaviours according to extracurricular sport participation was analysed by Chi-square tests.
Results
In boys, the use of internet for non-study was negatively associated with whole body BMC after adjustment for lean mass and moderate to vigorous PA (MVPA). In girls, the time spent studying was negatively associated with femoral neck BMC. Additional adjustment for lean mass slightly reduced the negative association between time spent studying and femoral neck BMC. The additional adjustment for MVPA did not change the results at this site. The percentage of girls having low femoral neck BMC was significantly smaller in those participating in osteogenic sports (≥ 3 h/week) than in the rest, independently of the cut-off selected for the time spent studying.
Conclusions
The use of internet for non-study (in boys) and the time spent studying (in girls) are negatively associated with whole body and femoral neck BMC, respectively. In addition, at least 3 h/week of extra-curricular osteogenic sports may help to counteract the negative association of time spent studying on bone health in girls.
doi:10.1186/1471-2458-12-971
PMCID: PMC3508981  PMID: 23148760
Bone health; Sedentary behaviours; Adolescents; Physical activity and extra-curricular participation in sports
6.  Physical Activity Attenuates the Effect of Low Birth Weight on Insulin Resistance in Adolescents 
Diabetes  2011;60(9):2295-2299.
OBJECTIVE
To examine whether physical activity influences the association between birth weight and insulin resistance in adolescents.
RESEARCH DESIGN AND METHODS
The study comprised adolescents who participated in two cross-sectional studies: the Healthy Lifestyle in Europe by Nutrition in Adolescence (HELENA) study (n = 520, mean age = 14.6 years) and the Swedish part of the European Youth Heart Study (EYHS) (n = 269, mean age = 15.6 years). Participants had valid data on birth weight (parental recall), BMI, sexual maturation, maternal education, breastfeeding, physical activity (accelerometry, counts/minute), fasting glucose, and insulin. Insulin resistance was assessed by homeostasis model assessment–insulin resistance (HOMA-IR). Maternal education level and breastfeeding duration were reported by the mothers.
RESULTS
There was a significant interaction of physical activity in the association between birth weight and HOMA-IR (logarithmically transformed) in both the HELENA study and the EYHS (P = 0.05 and P = 0.03, respectively), after adjusting for sex, age, sexual maturation, BMI, maternal education level, and breastfeeding duration. Stratified analyses by physical activity levels (below/above median) showed a borderline inverse association between birth weight and HOMA-IR in the low-active group (standardized β = −0.094, P = 0.09, and standardized β = −0.156, P = 0.06, for HELENA and EYHS, respectively), whereas no evidence of association was found in the high-active group (standardized β = −0.031, P = 0.62, and standardized β = 0.053, P = 0.55, for HELENA and EYHS, respectively).
CONCLUSIONS
Higher levels of physical activity may attenuate the adverse effects of low birth weight on insulin sensitivity in adolescents. More observational data, from larger and more powerful studies, are required to test these findings.
doi:10.2337/db10-1670
PMCID: PMC3161315  PMID: 21752955
7.  Television habits in relation to overweight, diet and taste preferences in European children: the IDEFICS study 
European Journal of Epidemiology  2012;27(9):705-715.
Early television exposure has been associated with various health outcomes including childhood obesity. This paper describes associations between patterns of television viewing, on one hand, and diet, taste preference and weight status, on the other, in European preschoolers and schoolchildren. The IDEFICS baseline survey was conducted at examination centers in Italy, Estonia, Cyprus, Belgium, Sweden, Germany, Hungary, and Spain. 15,144 children aged 2–9 completed the basic protocol, including anthropometry and parental questionnaires on their diets and television habits. A subsample of 1,696 schoolchildren underwent further sensory testing for fat and sweet taste preferences. Three dichotomous indicators described: children’s habitual television exposure time; television viewing during meals; and having televisions in their bedrooms. Based on these variables we investigated television habits in relation to overweight (IOTF) and usual consumption of foods high in fat and sugar. A possible role of taste preference in the latter association was tested in the sensory subgroup. All television indicators were significantly associated with increased risk of overweight, with odds ratios ranging from 1.21 to 1.30, in fully adjusted models. Children’s propensities to consume high-fat and high-sugar foods were positively and, in most analyses, monotonically associated with high-risk television behaviors. The associations between television and diet propensities were not explained by preference for added fat or sugar in test foods. To summarize, in addition to being more overweight, children with high-risk television behaviors may, independent of objectively measured taste preferences for fat and sugar, passively overconsume higher-fat and particularly higher-sugar diets.
doi:10.1007/s10654-012-9718-2
PMCID: PMC3486991  PMID: 22911022
Television; Diet; Taste preference; Childhood overweight
8.  Main characteristics and participation rate of European adolescents included in the HELENA study 
Archives of Public Health  2012;70(1):14.
Background
Participation rate and response rate are key issues in a cross sectional large-scale epidemiological study. The objective of this paper is to describe the study population and to evaluate participation and response rate as well as the key nutritional status variables in male and female adolescents involved in the HELENA study.
Methods
A multi-stage random cluster sampling with a target sample of 3000 adolescents aged [12.5 to 17.5] years, stratified for geographical location and age, was carried out. Information for participants and non-participants (NP) was compared, and participation and response rates to specific questionnaires were discussed.
Results
3,865 adolescents aged [12.5 to 17.5] years (1,845 females) participated in the HELENA study, of whom 1,076 (568 females) participated in the blood sampling. 3,528 (1,845 females) adolescents were finally kept for statistical analysis. Participation rates for the schools and classes differed importantly between countries. The participation rate of pupils within the participating classes also differed importantly between countries. Sex ratio, mean age and BMI were similar between NP and participating adolescents within each centre, and in the overall sample. For all the questionnaires included in the database, the response rate of questionnaires was high (more than 80% of questions were completed).
Conclusion
From this study it could be concluded that participation rate differed importantly between countries, though no bias could be identified when comparing the key study variables between participants and non-participants. Response rate for questionnaires was very high. Future studies investigating lifestyle and health in adolescents can optimize their methods when considering the opportunities and barriers observed in the HELENA study.
doi:10.1186/0778-7367-70-14
PMCID: PMC3490738  PMID: 22958310
9.  Analysis on the Time and Frequency Domains of the Acceleration in Front Crawl Stroke 
Journal of Human Kinetics  2012;32:109-120.
The swimming involves accelerations and decelerations in the swimmer’s body. Thus, the main objective of this study is to make a temporal and frequency analysis of the acceleration in front crawl swimming, regarding the gender and the performance. The sample was composed by 31 male swimmers (15 of high-level and 16 of low-level) and 20 female swimmers (11 of high-level and 9 of low-level). The acceleration was registered from the third complete cycle during eight seconds in a 25 meters maximum velocity test. A position transducer (200Hz) was used to collect the data, and it was synchronized to an aquatic camera (25Hz). The acceleration in the temporal (root mean square, minimum and maximum of the acceleration) and frequency (power peak, power peak frequency and spectral area) domains was calculated with Fourier analysis, as well as the velocity and the spectrums distribution in function to present one or more main peaks (type 1 and type 2). A one-way ANOVA was used to establish differences between gender and performance. Results show differences between genders in all the temporal domain variables (p<0.05) and only the Spectral Area (SA) in the frequency domain (p<0.05). Between gender and performance, only the Root Mean Square (RMS) showed differences in the performance of the male swimmers (p<0.05) and in the higher level swimmers, the Maximum (Max) and the Power Peak (PP) of the acceleration showed differences between both genders (p<0.05). These results confirms the importance of knowing the RMS to determine the efficiency of the swimmers regarding gender and performance level
doi:10.2478/v10078-012-0028-2
PMCID: PMC3590863  PMID: 23487001
swimming; Root Mean Square; power peak; power peak frequency; spectrum area
10.  Differences in Weight Status and Energy-Balance Related Behaviors among Schoolchildren across Europe: The ENERGY-Project 
PLoS ONE  2012;7(4):e34742.
Background
Current data on the prevalence of overweight and energy-balance behaviors among European children is necessary to inform overweight prevention interventions.
Methodology/Principal Findings
A school-based survey among 10–12 year old children was conducted in seven European countries using a standardized protocol. Weight, height, and waist circumference were measured; Engagement in physical activity, sedentary and dietary behaviors, and sleep duration were self-reported. Descriptive analyses were conducted, looking at differences according to country, gender, and parental education. 7234 children (52%girls; 11.6±0.7 years) participated. 25.8% and 5.4% of boys, and 21.8% and 4.1% of girls were overweight (including obese) and obese (according to International Obesity Task Force criteria), respectively. Higher prevalence of overweight/obesity was observed in Greece, Hungary, Slovenia and Spain than in Belgium, Netherlands and Norway. Large differences between countries were found in intakes of sugar-sweetened beverages, breakfast, active transport, TV and computer time. More favorable overweight status and behavior patterns were found in girls than boys and in children of higher educated parents than in children of lower educated parents.
Conclusions/Significance
High levels and striking differences in overweight status and potential risk behaviors were found among schoolchildren across Europe.
doi:10.1371/journal.pone.0034742
PMCID: PMC3338827  PMID: 22558098
11.  Do children and their parents eat a similar diet? Resemblance in child and parental dietary intake--systematic review and meta-analysis 
Objectives
Parents are believed having strong influence on children’s eating behaviors. However, previous findings on child-parent resemblance in dietary intakes are mixed. We systematically reviewed and meta-analyzed the association (correlations) based on published studies.
Methods
We searched related studies published since 1980 and found 24 studies meeting inclusion criteria for review and 15 for meta-regression analysis. We compared the associations between parent-child pairs, nutrients, over time, and by dietary assessment method.
Results
Most studies were based on small samples. Overall, they suggest a moderate or weak association, but findings varied remarkably. Our meta-analysis showed average Fisher’s transformed correlations were 0.20 (95% confidence interval 0.13–0.28) for fat (% energy); for energy, 0.21 (0.18, 0.24). The correlations varied by parent-child pairs, dietary assessment and countries. FFQ or mixed approaches yielded lower correlation than 24-hour recalls or food records. Child self-reported intakes showed weaker correlation, better methodology quality showed stronger correlation in fat intake (% energy), which also became weaker over time.
Conclusions
Overall, the resemblance is weak and it varied considerably across studies, nutrients, foods, parent-child pairs.
doi:10.1136/jech.2009.095901
PMCID: PMC3010265  PMID: 21051779
child; parent; diet; resemblance; association; review; meta-analysis
12.  Effects of Phosphorylation in Chlamydomonas Centrin Ser 167 
Calcium binding proteins  2006;1(2):108-114.
Centrin is a conserved calcium binding protein belonging to the EF-hand superfamily with two independent structural domains. This protein is found to be phosphorylated near the carboxyl terminal end. Our goal was to perform a novel comparative study of phosphorylated and unphosphorylated centrin by Fourier transform infrared (FT-IR) spectroscopy, two-dimensional correlation spectroscopy (2D-COS) analysis and differential scanning calorimetry (DSC). To achieve this goal, we have bacterially expressed, isolated, purified and phosphorylated centrin. We verified the extent of phosphorylation to be >97% for centrin by MALDI MS analysis and determined the absence of aggregated protein. The thermal denaturation temperature and ΔCp were determined to be Tm = 112.1 °C (ΔCp = 7.8 Kcal/mole/ΔC) and Tm = 111.0°C (ΔCp = 5.0 Kcal/mole/°C) for holo-centrin and phosphorylated centrin, respectively. We have also described the molecular dynamics leading up to the thermal denaturation of the protein: for holo-centrin the vibrational modes associated with the calcium binding sites aspartates and glutamates, loops then the arginines, followed by the structured backbone vibrational modes the α-helix at 1635 cm−1 then β-sheet and finally the more exposed α-helix at 1650 cm−1; while for phosphorylated centrin aspartate, glutamate and arginine, followed by the backbone associated vibrational modes α-helix (1650 cm−1), loop then the β-sheet (1633 cm−1) and finally the α-helix (1637 cm−1). Therefore, the effect on domain stability due to phosphorylation at Ser167 was observed in the loops as well as the α-helix at 1650 cm−1.
PMCID: PMC3232031  PMID: 22162668
calcium binding protein; phosphorylation; FT-IR Spectroscopy; differential scanning calorimetry; two dimensional correlation analysis; relative stability; conformational changes
13.  What do parents think about parental participation in school-based interventions on energy balance-related behaviours? a qualitative study in 4 countries 
BMC Public Health  2011;11:881.
Background
Overweight and obesity in youth has increased dramatically. Therefore, overweight prevention initiatives should start early in life and target modifiable energy balance-related behaviours. Parental participation is often advocated as important for school-based interventions, however, getting parents involved in school-based interventions appears to be challenging based on earlier intervention experiences. The purpose of this study was to get insight into the determinants of and perspectives on parental participation in school-interventions on energy balance-related behaviours (physical activity, healthy eating, sedentary behaviours) in parents of ten- to twelve-year olds in order to develop an effective parental module for school-based interventions concerning energy balance-related behaviours.
Methods
Four countries (Belgium, Hungary, Norway and Spain) conducted the focus group research based on a standardised protocol and a semi-structured questioning route. A variation in parental socio-economic status (SES) and parental school involvement was taken into account when recruiting the parents. The audio taped interviews were transcribed, and a qualitative content analysis of the transcripts was conducted in each country.
Results
Seventeen focus group interviews were conducted with a total of 92 parents (12 men, 80 women). Physical activity was considered to be a joint responsibility of school and parents, nutrition as parent's responsibility but supported by the school, and prevention of sedentary behaviours as parent's sole responsibility. Parents proposed interactive and practical activities together with their child as the best way to involve them such as cooking, food tasting, nutrition workshops, walking or cycling tours, sport initiations together with their child. Activities should be cheap, on a convenient moment, focused on their children and not on themselves, not tutoring, not theoretical, and school-or home-based.
Conclusions
Parents want to be involved in activities related to energy balance-related behaviours if this implies 'doing things together' with their child at school or at home.
doi:10.1186/1471-2458-11-881
PMCID: PMC3252293  PMID: 22112159
14.  Sequence and Role in Virulence of the Three Plasmid Complement of the Model Tumor-Inducing Bacterium Pseudomonas savastanoi pv. savastanoi NCPPB 3335 
PLoS ONE  2011;6(10):e25705.
Pseudomonas savastanoi pv. savastanoi NCPPB 3335 is a model for the study of the molecular basis of disease production and tumor formation in woody hosts, and its draft genome sequence has been recently obtained. Here we closed the sequence of the plasmid complement of this strain, composed of three circular molecules of 78,357 nt (pPsv48A), 45,220 nt (pPsv48B), and 42,103 nt (pPsv48C), all belonging to the pPT23A-like family of plasmids widely distributed in the P. syringae complex. A total of 152 coding sequences were predicted in the plasmid complement, of which 38 are hypothetical proteins and seven correspond to putative virulence genes. Plasmid pPsv48A contains an incomplete Type IVB secretion system, the type III secretion system (T3SS) effector gene hopAF1, gene ptz, involved in cytokinin biosynthesis, and three copies of a gene highly conserved in plant-associated proteobacteria, which is preceded by a hrp box motif. A complete Type IVA secretion system, a well conserved origin of transfer (oriT), and a homolog of the T3SS effector gene hopAO1 are present in pPsv48B, while pPsv48C contains a gene with significant homology to isopentenyl-diphosphate delta-isomerase, type 1. Several potential mobile elements were found on the three plasmids, including three types of MITE, a derivative of IS801, and a new transposon effector, ISPsy30. Although the replication regions of these three plasmids are phylogenetically closely related, their structure is diverse, suggesting that the plasmid architecture results from an active exchange of sequences. Artificial inoculations of olive plants with mutants cured of plasmids pPsv48A and pPsv48B showed that pPsv48A is necessary for full virulence and for the development of mature xylem vessels within the knots; we were unable to obtain mutants cured of pPsv48C, which contains five putative toxin-antitoxin genes.
doi:10.1371/journal.pone.0025705
PMCID: PMC3191145  PMID: 22022435
15.  Reliability and validity of the Adolescent Stress Questionnaire in a sample of European adolescents - the HELENA study 
BMC Public Health  2011;11:717.
Background
Since stress is hypothesized to play a role in the etiology of obesity during adolescence, research on associations between adolescent stress and obesity-related parameters and behaviours is essential. Due to lack of a well-established recent stress checklist for use in European adolescents, the study investigated the reliability and validity of the Adolescent Stress Questionnaire (ASQ) for assessing perceived stress in European adolescents.
Methods
The ASQ was translated into the languages of the participating cities (Ghent, Stockholm, Vienna, Zaragoza, Pecs and Athens) and was implemented within the HELENA cross-sectional study. A total of 1140 European adolescents provided a valid ASQ, comprising 10 component scales, used for internal reliability (Cronbach α) and construct validity (confirmatory factor analysis or CFA). Contributions of socio-demographic (gender, age, pubertal stage, socio-economic status) characteristics to the ASQ score variances were investigated. Two-hundred adolescents also provided valid saliva samples for cortisol analysis to compare with the ASQ scores (criterion validity). Test-retest reliability was investigated using two ASQ assessments from 37 adolescents.
Results
Cronbach α-values of the ASQ scales (0.57 to 0.88) demonstrated a moderate internal reliability of the ASQ, and intraclass correlation coefficients (0.45 to 0.84) established an insufficient test-retest reliability of the ASQ. The adolescents' gender (girls had higher stress scores than boys) and pubertal stage (those in a post-pubertal development had higher stress scores than others) significantly contributed to the variance in ASQ scores, while their age and socio-economic status did not. CFA results showed that the original scale construct fitted moderately with the data in our European adolescent population. Only in boys, four out of 10 ASQ scale scores were a significant positive predictor for baseline wake-up salivary cortisol, suggesting a rather poor criterion validity of the ASQ, especially in girls.
Conclusions
In our European adolescent sample, the ASQ had an acceptable internal reliability and construct validity and the adolescents' gender and pubertal stage systematically contributed to the ASQ variance, but its test-retest reliability and criterion validity were rather poor. Overall, the utility of the ASQ for assessing perceived stress in adolescents across Europe is uncertain and some aspects require further examination.
doi:10.1186/1471-2458-11-717
PMCID: PMC3188495  PMID: 21943341
16.  Determination of the melon chloroplast and mitochondrial genome sequences reveals that the largest reported mitochondrial genome in plants contains a significant amount of DNA having a nuclear origin 
BMC Genomics  2011;12:424.
Background
The melon belongs to the Cucurbitaceae family, whose economic importance among vegetable crops is second only to Solanaceae. The melon has a small genome size (454 Mb), which makes it suitable for molecular and genetic studies. Despite similar nuclear and chloroplast genome sizes, cucurbits show great variation when their mitochondrial genomes are compared. The melon possesses the largest plant mitochondrial genome, as much as eight times larger than that of other cucurbits.
Results
The nucleotide sequences of the melon chloroplast and mitochondrial genomes were determined. The chloroplast genome (156,017 bp) included 132 genes, with 98 single-copy genes dispersed between the small (SSC) and large (LSC) single-copy regions and 17 duplicated genes in the inverted repeat regions (IRa and IRb). A comparison of the cucumber and melon chloroplast genomes showed differences in only approximately 5% of nucleotides, mainly due to short indels and SNPs. Additionally, 2.74 Mb of mitochondrial sequence, accounting for 95% of the estimated mitochondrial genome size, were assembled into five scaffolds and four additional unscaffolded contigs. An 84% of the mitochondrial genome is contained in a single scaffold. The gene-coding region accounted for 1.7% (45,926 bp) of the total sequence, including 51 protein-coding genes, 4 conserved ORFs, 3 rRNA genes and 24 tRNA genes. Despite the differences observed in the mitochondrial genome sizes of cucurbit species, Citrullus lanatus (379 kb), Cucurbita pepo (983 kb) and Cucumis melo (2,740 kb) share 120 kb of sequence, including the predicted protein-coding regions. Nevertheless, melon contained a high number of repetitive sequences and a high content of DNA of nuclear origin, which represented 42% and 47% of the total sequence, respectively.
Conclusions
Whereas the size and gene organisation of chloroplast genomes are similar among the cucurbit species, mitochondrial genomes show a wide variety of sizes, with a non-conserved structure both in gene number and organisation, as well as in the features of the noncoding DNA. The transfer of nuclear DNA to the melon mitochondrial genome and the high proportion of repetitive DNA appear to explain the size of the largest mitochondrial genome reported so far.
doi:10.1186/1471-2164-12-424
PMCID: PMC3175227  PMID: 21854637
17.  Synergic effect of salinity and zinc stress on growth and photosynthetic responses of the cordgrass, Spartina densiflora 
Journal of Experimental Botany  2011;62(15):5521-5530.
Spartina densiflora is a C4 halophytic species that has proved to have a high invasive potential which derives from its physiological plasticity to environmental factors, such as salinity. It is found in coastal marshes of south-west Spain, growing over sediments with between 1 mmol l−1 and 70 mmol l−1 zinc. A glasshouse experiment was designed to investigate the synergic effect of zinc from 0 mmol l−1 to 60 mmol l−1 at 0, 1, and 3% NaCl on the growth and the photosynthetic apparatus of S. densiflora by measuring chlorophyll fluorescence parameters and gas exchange, and its recovery after removing zinc. Antioxidant enzyme activities and total zinc, sodium, calcium, iron, magnesium, manganese, phosphorus, potassium, and nitrogen concentrations were also determined. Spartina densiflora showed the highest growth at 1 mmol l−1 zinc and 1% NaCl after 90 d of treatment; this enhanced growth was supported by the measurements of net photosynthetic rate (A). Furthermore, there was a stimulatory effect of salinity on accumulation of zinc in tillers of this species. Zinc concentrations >1 mmol l−1 reduced growth of S. densiflora, regardless of salinity treatments. This declining growth may be attributed to a decrease in A caused by diffusional limitation of photosynthesis, owing to the modification of the potassium/calcium ratio. Also, zinc and salinity had a marked overall effect on the photochemical (photosystem II) apparatus, partially mediated by the accumulation of H2O2 and subsequent oxidative damage. However, salinity favoured the recovery of the photosynthetic apparatus to the toxic action of zinc, and enhanced the nutrient uptake.
doi:10.1093/jxb/err234
PMCID: PMC3223049  PMID: 21841175
Antioxidant enzyme activities; chlorophyll fluorescence; cordgrass; gas exchange; growth; salinity; synergic effect; zinc
18.  Using the intervention mapping protocol to develop a community-based intervention for the prevention of childhood obesity in a multi-centre European project: the IDEFICS intervention 
Background
The prevalence of childhood obesity has increased during the past decades and is now considered an urgent public health problem. Although stabilizing trends in obesity prevalence have been identified in parts of Europe, preventive efforts in children are still needed. Using the socio-ecological approach as the underlying theoretical perspective, the IDEFICS project aimed to develop, implement and evaluate a community-based intervention for the prevention of childhood obesity in eight European countries. The aim of the present manuscript was to describe the content and developmental process of the IDEFICS intervention.
Methods
The intervention mapping protocol (IMP) was used to develop the community-based intervention for the prevention of childhood obesity in 3 to 10 years old children. It is a theory- and evidence-based tool for the structured planning and development of health promotion programs that requires the completion of six different steps. These steps were elaborated by two coordinating centers and discussed with the other participating centers until agreement was reached. Focus group research was performed in all participating centers to provide an informed basis for intervention development.
Results
The application of the IMP resulted in an overall intervention framework with ten intervention modules targeting environmental and personal factors through the family, the school and the community. The summary results of the focus group research were used to inform the development of the overall intervention. The cultural adaptation of the overall intervention was realised by using country specific focus group results. The need for cultural adaptation was considered during the entire process to improve program adoption and implementation. A plan was developed to evaluate program effectiveness and quality of implementation.
Conclusions
The IDEFICS project developed a community-based intervention for the prevention of childhood obesity by using to the intervention mapping heuristic. The IDEFICS intervention consists of a general and standardized intervention framework that allows for cultural adaptation to make the intervention feasible and to enhance deliverability in all participating countries. The present manuscript demonstrates that the development of an intervention is a long process that needs to be done systematically. Time, human resources and finances need to be planned beforehand to make interventions evidence-based and culturally relevant.
doi:10.1186/1479-5868-8-82
PMCID: PMC3169445  PMID: 21806806
19.  Clustering patterns of physical activity, sedentary and dietary behavior among European adolescents: The HELENA study 
BMC Public Health  2011;11:328.
Background
Evidence suggests possible synergetic effects of multiple lifestyle behaviors on health risks like obesity and other health outcomes. A better insight in the clustering of those behaviors, could help to identify groups who are at risk in developing chronic diseases. This study examines the prevalence and clustering of physical activity, sedentary and dietary patterns among European adolescents and investigates if the identified clusters could be characterized by socio-demographic factors.
Methods
The study comprised a total of 2084 adolescents (45.6% male), from eight European cities participating in the HELENA (Healthy Lifestyle in Europe by Nutrition in Adolescence) study. Physical activity and sedentary behavior were measured using self-reported questionnaires and diet quality was assessed based on dietary recall. Based on the results of those three indices, cluster analyses were performed. To identify gender differences and associations with socio-demographic variables, chi-square tests were executed.
Results
Five stable and meaningful clusters were found. Only 18% of the adolescents showed healthy and 21% unhealthy scores on all three included indices. Males were highly presented in the cluster with high levels of moderate to vigorous physical activity (MVPA) and low quality diets. The clusters with low levels of MVPA and high quality diets comprised more female adolescents. Adolescents with low educated parents had diets of lower quality and spent more time in sedentary activities. In addition, the clusters with high levels of MVPA comprised more adolescents of the younger age category.
Conclusion
In order to develop effective primary prevention strategies, it would be important to consider multiple health indices when identifying high risk groups.
doi:10.1186/1471-2458-11-328
PMCID: PMC3112135  PMID: 21586158
20.  Risk of injury for bicycling on cycle tracks versus in the street 
Injury Prevention  2011;17(2):131-135.
Most individuals prefer bicycling separated from motor traffic. However, cycle tracks (physically separated bicycle-exclusive paths along roads, as found in The Netherlands) are discouraged in the USA by engineering guidance that suggests that facilities such as cycle tracks are more dangerous than the street. The objective of this study conducted in Montreal (with a longstanding network of cycle tracks) was to compare bicyclist injury rates on cycle tracks versus in the street. For six cycle tracks and comparable reference streets, vehicle/bicycle crashes and health record injury counts were obtained and use counts conducted. The relative risk (RR) of injury on cycle tracks, compared with reference streets, was determined. Overall, 2.5 times as many cyclists rode on cycle tracks compared with reference streets and there were 8.5 injuries and 10.5 crashes per million bicycle-kilometres. The RR of injury on cycle tracks was 0.72 (95% CI 0.60 to 0.85) compared with bicycling in reference streets. These data suggest that the injury risk of bicycling on cycle tracks is less than bicycling in streets. The construction of cycle tracks should not be discouraged.
doi:10.1136/ip.2010.028696
PMCID: PMC3064866  PMID: 21307080
Bicycle; engineering; environment; public health; safe community
21.  Relationship between self-reported dietary intake and physical activity levels among adolescents: The HELENA study 
Background
Evidence suggests possible synergetic effects of multiple lifestyle behaviors on health risks like obesity and other health outcomes. Therefore it is important to investigate associations between dietary and physical activity behavior, the two most important lifestyle behaviors influencing our energy balance and body composition. The objective of the present study is to describe the relationship between energy, nutrient and food intake and the physical activity level among a large group of European adolescents.
Methods
The study comprised a total of 2176 adolescents (46.2% male) from ten European cities participating in the HELENA (Healthy Lifestyle in Europe by Nutrition in Adolescence) study. Dietary intake and physical activity were assessed using validated 24-h dietary recalls and self-reported questionnaires respectively. Analyses of covariance (ANCOVA) were used to compare the energy and nutrient intake and the food consumption between groups of adolescents with different physical activity levels (1st to 3rd tertile).
Results
In both sexes no differences were found in energy intake between the levels of physical activity. The most active males showed a higher intake of polysaccharides, protein, water and vitamin C and a lower intake of saccharides compared to less active males. Females with the highest physical activity level consumed more polysaccharides compared to their least active peers. Male and female adolescents with the highest physical activity levels, consumed more fruit and milk products and less cheese compared to the least active adolescents. The most active males showed higher intakes of vegetables and meat, fish, eggs, meat substitutes and vegetarian products compared to the least active ones. The least active males reported the highest consumption of grain products and potatoes. Within the female group, significantly lower intakes of bread and cereal products and spreads were found for those reporting to spend most time in moderate to vigorous physical activity. The consumption of foods from the remaining food groups, did not differ between the physical activity levels in both sexes.
Conclusion
It can be concluded that dietary habits diverge between adolescents with different self-reported physical activity levels. For some food groups a difference in intake could be found, which were reflected in differences in some nutrient intakes. It can also be concluded that physically active adolescents are not always inclined to eat healthier diets than their less active peers.
doi:10.1186/1479-5868-8-8
PMCID: PMC3045277  PMID: 21294914
22.  EuropeaN Energy balance Research to prevent excessive weight Gain among Youth (ENERGY) project: Design and methodology of the ENERGY cross-sectional survey 
BMC Public Health  2011;11:65.
Background
Obesity treatment is by large ineffective long term, and more emphasis on the prevention of excessive weight gain in childhood and adolescence is warranted. To inform energy balance related behaviour (EBRB) change interventions, insight in the potential personal, family and school environmental correlates of these behaviours is needed. Studies on such multilevel correlates of EBRB among schoolchildren in Europe are lacking. The ENERGY survey aims to (1) provide up-to-date prevalence rates of measured overweight, obesity, self-reported engagement in EBRBs, and objective accelerometer-based assessment of physical activity and sedentary behaviour and blood-sample biomarkers of metabolic function in countries in different regions of Europe, (2) to identify personal, family and school environmental correlates of these EBRBs. This paper describes the design, methodology and protocol of the survey.
Method/Design
A school-based cross-sectional survey was carried out in 2010 in seven different European countries; Belgium, Greece, Hungary, the Netherlands, Norway, Slovenia, and Spain. The survey included measurements of anthropometrics, child, parent and school-staff questionnaires, and school observations to measure and assess outcomes (i.e. height, weight, and waist circumference), EBRBs and potential personal, family and school environmental correlates of these behaviours including the social-cultural, physical, political, and economic environmental factors. In addition, a selection of countries conducted accelerometer measurements to objectively assess physical activity and sedentary behaviour, and collected blood samples to assess several biomarkers of metabolic function.
Discussion
The ENERGY survey is a comprehensive cross-sectional study measuring anthropometrics and biomarkers as well as assessing a range of EBRBs and their potential correlates at the personal, family and school level, among 10-12 year old children in seven European countries. This study will result in a unique dataset, enabling cross country comparisons in overweight, obesity, risk behaviours for these conditions as well as the correlates of engagement in these risk behaviours.
doi:10.1186/1471-2458-11-65
PMCID: PMC3044658  PMID: 21281466
23.  Breast-Feeding Modulates the Influence of the Peroxisome Proliferator–Activated Receptor-γ (PPARG2) Pro12Ala Polymorphism on Adiposity in Adolescents 
Diabetes Care  2009;33(1):190-196.
OBJECTIVE
The peroxisome proliferator–activated receptor-γ2 (PPARG2) Pro12Ala polymorphism has been associated with a higher BMI and a lower risk of type 2 diabetes in adulthood. The association between adiposity and PPARG variants can be influenced by environmental factors such as early growth, dietary fat, and (as recently shown) breast-feeding. The objectives of this study were to assess 1) the influence of the PPARG2 Pro12Ala polymorphism on adiposity markers in adolescents and 2) a possible modulating effect of breast-feeding on these associations.
RESEARCH DESIGN AND METHODS
Data on breast-feeding duration, BMI, and genotypes for the Pro12Ala polymorphism were available for 945 adolescents (mean age 14.7 years). The breast-feeding duration was obtained from parental records. We measured weight, height, waist circumference, and six skinfold thicknesses.
RESULTS
No significant associations between the Pro12Ala polymorphism and any of the above-mentioned anthropometric parameters were found. There were significant interactions between the PPARG2 Pro12Ala polymorphism and breast-feeding with regard to adiposity measurements (all adjusted P < 0.05). Indeed, in children who had not been breast-fed, Ala12 allele carriers had higher adiposity parameters (e.g., Δ BMI +1.88 kg/m2, adjusted for age, sex, and center, P = 0.007) than Pro12Pro adolescents. In contrast, in breast-fed subjects, there was no significant difference between Ala12 allele carriers and Pro12Pro children in terms of adiposity measurements, whatever the duration of breast-feeding.
CONCLUSIONS
Breast-feeding appears to counter the deleterious effect of the PPARG2 Pro12Ala polymorphism on anthropometric parameters in adolescents.
doi:10.2337/dc09-1459
PMCID: PMC2797971  PMID: 19846795
24.  Fate of a Pseudomonas savastanoi pv. savastanoi Type III Secretion System Mutant in Olive Plants (Olea europaea L.)▿†  
Applied and Environmental Microbiology  2010;76(11):3611-3619.
Pseudomonas savastanoi pv. savastanoi strain NCPPB 3335 is a model bacterial pathogen for studying the molecular basis of disease production in woody hosts. We report the sequencing of the hrpS-to-hrpZ region of NCPPB 3335, which has allowed us to determine the phylogenetic position of this pathogen with respect to previously sequenced Pseudomonas syringae hrp clusters. In addition, we constructed a mutant of NCPPB 3335, termed T3, which carries a deletion from the 3′ end of the hrpS gene to the 5′ end of the hrpZ operon. Despite its inability to multiply in olive tissues and to induce tumor formation in woody olive plants, P. savastanoi pv. savastanoi T3 can induce knot formation on young micropropagated olive plants. However, the necrosis and formation of internal open cavities previously reported in knots induced by the wild-type strain were not observed in those induced by P. savastanoi pv. savastanoi T3. Tagging of P. savastanoi pv. savastanoi T3 with green fluorescent protein (GFP) allowed real-time monitoring of its behavior on olive plants. In olive plant tissues, the wild-type strain formed aggregates that colonized the intercellular spaces and internal cavities of the hypertrophic knots, while the mutant T3 strain showed a disorganized distribution within the parenchyma of the knot. Ultrastructural analysis of knot sections revealed the release of extensive outer membrane vesicles from the bacterial cell surface of the P. savastanoi pv. savastanoi T3 mutant, while the wild-type strain exhibited very few vesicles. This phenomenon has not been described before for any other bacterial phytopathogen during host infection.
doi:10.1128/AEM.00133-10
PMCID: PMC2876471  PMID: 20363790
25.  Genome-wide BAC-end sequencing of Cucumis melo using two BAC libraries 
BMC Genomics  2010;11:618.
Background
Although melon (Cucumis melo L.) is an economically important fruit crop, no genome-wide sequence information is openly available at the current time. We therefore sequenced BAC-ends representing a total of 33,024 clones, half of them from a previously described melon BAC library generated with restriction endonucleases and the remainder from a new random-shear BAC library.
Results
We generated a total of 47,140 high-quality BAC-end sequences (BES), 91.7% of which were paired-BES. Both libraries were assembled independently and then cross-assembled to obtain a final set of 33,372 non-redundant, high-quality sequences. These were grouped into 6,411 contigs (4.5 Mb) and 26,961 non-assembled BES (14.4 Mb), representing ~4.2% of the melon genome. The sequences were used to screen genomic databases, identifying 7,198 simple sequence repeats (corresponding to one microsatellite every 2.6 kb) and 2,484 additional repeats of which 95.9% represented transposable elements. The sequences were also used to screen expressed sequence tag (EST) databases, revealing 11,372 BES that were homologous to ESTs. This suggests that ~30% of the melon genome consists of coding DNA. We observed regions of microsynteny between melon paired-BES and six other dicotyledonous plant genomes.
Conclusion
The analysis of nearly 50,000 BES from two complementary genomic libraries covered ~4.2% of the melon genome, providing insight into properties such as microsatellite and transposable element distribution, and the percentage of coding DNA. The observed synteny between melon paired-BES and six other plant genomes showed that useful comparative genomic data can be derived through large scale BAC-end sequencing by anchoring a small proportion of the melon genome to other sequenced genomes.
doi:10.1186/1471-2164-11-618
PMCID: PMC3091759  PMID: 21054843

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