Objectives: To evaluate the effect of gender and puberty on cardiovascular risk factors (CVRF) in obese children and adolescents.Methods: One thousand four hundred and nine obese patients  were studied. Subjects were stratified according to Tanner pubertal staging and age into prepubertal ≤ and >6 ys (G1 and G2), pubertal stage 2–3 (G3), and pubertal stage 4–5 (G4). Waist circumference (WC), systolic and diastolic blood pressure , fasting plasma glucose, insulin, post Oral Glucose Tolerance Test glucose and insulin, and lipids were evaluated. Insulin resistance was evaluated by HOMA index. Patients with no CVRF were considered metabolically healthy (MHO).Results: The percentage of MHO patients was 59.8% in G1 while was consistently around 30% in the other groups. WC was more frequently abnormal in G2 males. Pubertal progression was associated with a decrease in WC abnormalities. SP was more frequently abnormal in G4 males and pubertal progression was associated with higher prevalence of abnormal SP in males. Pubertal progression was associated with an increase in hypertension rate in both sexes. HOMA was more frequently abnormal in G2 and G3 females. HDL, LDL, and TG were more frequently abnormal in G2 females. Dyslipidemia rate was higher in G2 females. Pubertal progression was associated with higher prevalence of abnormal HDL in males.Conclusions: Sex and pubertal status influence the frequency of abnormalities of CVRF in obese children and adolescents. CVRF are already present in prepubertal age. Identifying patients with higher risk of metabolic complications is important to design targeted and effective prevention strategies. Childhood obesity is one of the major public health problems. In recent decades, the prevalence of obesity has considerably increased worldwide due to sedentary lifestyle and high calories diets –3. A recThe influence of puberty and sex on obesity related cardiovascular risk factors (CVRF) has been shown in a number of studies –9. In adThe aim of this study was to evaluate the effect of sex and pubertal maturation on CVRF in a large group of children and adolescents with obesity.Data were collected from 1,409 children and adolescents with obesity referred to our Endocrine Unit between 2000 and 2016. The study was approved by our ethic committee and written informed consent was obtained from the parents or legal guardians for all patients. All patients included in the study had simple obesity, and possible organic or genetic causes were excluded.In all subjects height, weight, pubertal status [according to Tanner and Whitehouse ], waist Morning serum concentrations of glucose (Gly), insulin (Ins), total cholesterol (tCHO), HDL, LDL, triglycerides (TG) were evaluated in all subjects, between 08.00 and 09.00 h, after an overnight fast, in supine position. On the same day, an oral glucose tolerance test  was performed. Homeostatic model assessment  was used to evaluate insulin resistance.WC was considered normal if <90th percentile , 25. HypChildren with normal SP, DP, Gly, Gly OGTT, Ins, Ins OGTT, HOMA, tCHO, HDL, LDL, and TG were considered metabolically healthy obese patients (MHO), while children with one or more abnormal factor were considered metabolically unhealthy obese patients (MUO).Patients were analyzed as a single group as well as after subdivision into 4 groups, according to their pubertal status and age: prepubertal ≤6 years (group 1), prepubertal >6 years (group 2), pubertal status 2–3 (group 3), pubertal status 4–5 (group 4).Serum glucose was determined by the glucose oxidases method . Serum insulin concentrations were measured using reagents provided by Siemens . tCHO, LDL, HDL, and TG were measured by standard routine methods.post-hoc multiple comparison, according to the type of variables. When age distribution was not similar between males and females, and when BMI-SDS distribution was not similar between males and females and between pubertal groups, age- or BMI-SDS-adjusted ANCOVA followed by Bonferroni's test for post-hoc multiple comparison and logistic regression were used to compare characteristics among the study groups, according to the type of variables. Bonferroni correction was applied when appropriate. P ≤ 0.01 was considered significant. All statistical calculations were performed using STATISTICA version 9.1 software  and Graph Pad Prism version 6 .Distribution of the data was evaluated using Kolmogorov–Smirnov test. Data were presented as median and interquartile range (IQR) or count and percentage. Comparisons between 2 groups were performed using the Mann-Whitney U-test or Chi-Squared test, according to the type of variables. Comparisons between multiple groups were performed using Chi-Squared test or Kruskal-Wallis test followed by Dunn's test for Clinical and biochemical characteristics of the patients are summarized in Sex differences in CVRF in all patients are reported in Pubertal status differences in CVRF in all patients are reported in Sex differences in CVRF in patients subdivided according to their pubertal status are reported in Pubertal status differences in CVRF in patients subdivided according to sex are reported in The percentage of MHO patients was higher in group 1, and WCp was more frequently abnormal in group 2, regardless of sex. Among males, SP was less frequently abnormal in group 2.In summary:WC was more frequently abnormal in group 2 males. Pubertal progression was associated with a decrease in WC abnormalities.SP was more frequently abnormal in group 4 males and pubertal progression was associated with higher prevalence of abnormal SP in males. Pubertal progression was associated with an increase in hypertension rate in both males and female.The prevalence of Gly abnormalities was similar between males and females, at any pubertal stage considered. Conversely, HOMA was more frequently abnormal in groups 2 and 3 females compared to males.HDL, LDL, and TG were more frequently abnormal in group 2 females. Dyslipidemia rate was higher in group 2 females. Pubertal progression was associated with higher prevalence of abnormal HDL in males.Pubertal progression was not associated with change in MHO rate, but MHO rate was higher in group 1.The prevalence of childhood obesity and its complications are increasing worldwide, and, therefore, it is mandatory to recognize high risk categories on which to focus prevention interventions –3. ObesiIn our study, WCp, SP, and Gly were higher in boys while Ins, Ins OGTT and HOMA were higher in girls. In order to rule out a possible effect of puberty, we evaluated the effect of sex after subdividing the group according to pubertal status. We confirmed that WCp and SP were higher in group 2 and 4 boys, while Ins, Ins OGTT and HOMA were higher in group 2 and 3 girls. Moreover, group 2 girls had higher LDL and TG levels. No sex differences were found in prepubertal patients ≤6 years. We are aware that some result of this analysis, although statistically significant, may not always reflect a biological difference. For example, 5 mmHg difference in SP between boys and girls is not clinically relevant. However, when we looked for sex differences in the prevalence of CVRF abnormalities, we confirmed a higher prevalence of abnormal WC in group 2 boys, SP and hypertension in group 4 boys and HOMA, LDL, HDL, TG and dyslipidimia in group 2. A number of studies have shown, both in the general population and in subjects with obesity, an adverse cardiometabolic profile in boys , 35–37, The physiological changes of puberty driven by the rising secretion of sex hormones are accompanied by changes in body composition, fuel metabolism, lipids levels, blood pressure and reduced insulin sensitivity –17, 44. The cross-sectional study design is a clear limitation of the study. However, the large sample size allowed to show significant differences in the prevalence and severity of CVRF related to age, sex and pubertal status. Only few studies have evaluated specifically the effect of sex and puberty on CVRF in the pediatric population excluding possible confounding factors , 35, 36.In conclusion, our study confirms that sex and pubertal status influence the prevalence and severity of CVRF in children and adolescents with obesity. CVRF are already present in prepubertal age. Identifying patients with higher risk of metabolic complications is important to design targeted and effective prevention strategies.The datasets generated for this study are available on request to the corresponding author.This study was carried out in accordance with the recommendations of Independent Ethical Committee of the University of Cagliari with written informed consent from all subjects. All subjects gave written informed consent in accordance with the Declaration of Helsinki. The protocol was approved by the Independent Ethical Committee of the University of Cagliari, Italy.CG carried out the analyses, drafted the initial manuscript, and reviewed and revised the manuscript. AI, LC, SP, and SC designed the data collection instruments, collected data, and reviewed and revised the manuscript. SL conceptualized and designed the study, coordinated and supervised data collection, and critically reviewed the manuscript for important intellectual content. CG, AI, LC, SP, SC, and SL approved the final manuscript as submitted and agree to be accountable for all aspects of the work.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Microphysogobio rapidus (Cyprinidae), we surveyed a total of 79 sites from the historic records (20 sites) plus additional sites (59 sites) with good habitat conditions, analyzed their sites, and compared them with historic recorded sites to reveal the factors of extinction threats and causes. We found only eight out of 79 sites in the Nam River areas. The habitats were greatly reduced and restricted compared with the historic sites, which mainly cause from habitat modification, such as various types of river renovations at the main stream and tributary streams of the Nakdong River. The present habitats are higher water temperature and more number of fish species than the absent ones, but conductivity, total nitrogen, and number of weir are lower. In addition, the present sites are lower low velocity at pool and higher mean substrate at pool. From this study, we suggest that maintaining good water quality and preventing anthropogenic impacts greatly aid conservation of the M. rapidus in South Korea.To investigate distribution, habitat characteristics, and current conservation status of the endangered endemic species, rapid small gudgeon  Global biological diversity is rapidly diminishing as a result of human activity  was designated as an Endangered Wildlife Category I endemic species by the Ministry of Environment in 2012  20 times and a scoop net  for 30 min in a 100 m transect at each site. After identification, fish were released at their original capture site. The collected fish were identified according to Kim et al.  along the Nam River in Sancheong-gun and one site (St. 74) along the Deokcheon River in Jinju-si, where M. rapidus was most prevalent, according to the distribution survey; one site along the Yeong River in Sangju-si (St. 33) and one site along the Gam Stream in Gumi-si (St. 42), where M. rapidus used to be most prevalent but is no longer observed; and three sites, including the Yeongcheon River in Jinju-si (St. 77), which were previously preferred M. rapidus habitats but have recently been developed  was measured using a tachometer  about 5 cm above the stream bed. The water depth (WD) and bottom substrate were measured using a tape measure. The mean bottom substrate size (MS) was obtained by calculating the average of the major axes of 10 randomly selected sand, gravel, pebble, and cobble pieces from every 1 m2 at each site. The classification of the bottom substrate followed Cummins , while the other set included Banbyeon Stream in Andong-si (St. 18), Naeseong Stream in Yeocheon-gun (St. 24), Wi Stream in Uiseong-gun (St. 41), Hoe Stream in Goryeong-gun (St. 50), Miryang River in Miryang-si (St. 59), and three sites where M. rapidus did not appear during field work . The river width (RW) and water width (WW), which are the hydrological environmental factors of habitats, were measured using a digital range finder , and the altitude (AL) was found using Google Earth data. The physicochemical environmental factors, dissolved oxygen (DO), conductivity (CON), pH, water temperature (WT), suspended solids (SS), total nitrogen (TN), and total phosphorus (TP) were obtained from water quality monitoring network data (2004–2013) from the Water Environmental Information System of the National Institute of Environmental Research. The number of fish species (NFS) was measured as a biological factor, while the number of weirs (NW) was counted as another factor.The hydrological, physicochemical, biological, and other confounding factors influencing fish habitat were compared between two sets of eight sites selected from the whole 79 sites in the survey. The first set of sites included the Im Stream (St. 63), Yang Stream (St. 67), Nam River  in Sancheong-gun, and three sites where many M. rapidus was previously present but is now absent. Then, second analysis used the same methods to determine whether there was any difference between three presence and three absence sites in terms of hydrological data (6 variables) collected from 10 equal parts of each rapid and pool at every site . Before conducting the analysis, a normal distribution of environmental factor variables  was measured. The variables without a normal distribution were included in the analysis after confirming normality through data transformation.The software package SPSS  was used to analyze extinction threats and causes. We conducted two separate analyses because we measured different environmental variables at different places within sites. First, a one-way ANOVA was performed to compare the physicochemical, hydrological, and biological data (12 variables) between the eight presence and eight absence sites where M. rapidus. The PCA was performed to reduce the data to independent PCs that summarized most of the variance of the original variables . Then the second PCA was carried with 6 variables. The mean scores of the data for the variables were used, and the Equamax method was used to rotate the PC factor loadings. Then the PC scores for each component were compared between the presence and absence sites.After that, two separate principal component analysis (PCA) were carried out to estimate the environmental factors that most influenced the habitat conditions of M. rapidus presence sites (three sites) are in the Wi Stream and Nam River, followed by the main stream of the Nakdong, Yeong, Deokcheon River, and Yang Stream (two sites). Meanwhile, the Banbyeon, Naeseong, Gam, Hoe, Nabul, Daegok, Yeongcheon, and Miryang rivers each had one site . The LV and MS at pools were significantly different in test for the rapids and pool areas (10 for each presence and absence site) (P < .05).Of the 75 sites, the habitats of F1,14 = 7.094, p = .019; F1,58 = 15.150, p < .001; Four PCs with eigenvalues >1.0 were extracted from the PCA and applied to 10 environmental variables . The PC3M. rapidus from 1996 to 2008 with the current presence sites . Various habitat characteristics, including available shelter, spawning grounds, and prey, are required to accommodate many different species of fishes. The sites where M. rapidus was absent have had many flood damage rehabilitation and river renovation projects since 2000. Since 2010, the Four Major Rivers Project has led to the loss of riverbed materials and rising water levels at tributary streams, up to 6 m at the main stream of the Nakdong River. Such projects have transformed many habitats into lentic ecosystems, decreasing the number of rapids-preferred and benthic fish species, increasing the number nekton fishes , which helped to maintain the natural state of the river and retain critical fish habitat, resulting in a high NFS. In addition, CON and TN at the presence sites were also significantly different from those at the absence sites (P < .01). CON generally increased proportionally with the number of ionic contaminants. Absence sites tended to be in or near large cities or other areas with inflow of pollutants from point and non-point sources (e.g. farmlands and stables), which are likely to increase CON. On the other hand, clean water in the Nam River system was provided all year round at the presence sites, which likely results in low CON. An et al.  revealed that the extent of occurrence and area of occupancy were less than 5000 km2 and 500 km2, respectively. This indicates that the extent or quality of habitats and number of areas of M. rapidus have decreased compared to previous findings, and thus this species can be considered to qualify for the endangered B2ab  classification of the Red List, meaning that it is critically endangered in the wild. A previous study , and the designation of the coastal zone from St. 63 to St. 64 (which is estimated to have the largest population) as a legally protected area. In addition, based on the confirmed distribution and habitat features, conservation studies using continuous monitoring, fundamental ecological research, and genetic diversity analyses are required.The presence distribution survey indicated that the Deokcheon River group in the Nakdong River system had few downstream populations due to the rehabilitation projects and weir construction. However, it was confirmed through continuous monitoring that the population of


In the group supplemented with 10 mg-Zn/day, there was an increase in reactance values (p = 0.036) and PA (p = 0.002), in addition to vector displacement (p < 0.001) in relation to the confidence ellipses. An increase in serum zinc concentration was found (p < 0.001) in all three groups. Whit this, the supplementation with 10 mg-Zn/day promotes changes in the integrity of the cell membrane associated with the increase in the cellular mass of healthy children.The parameters derived from bioelectrical impedance, phase angle (PA) and bioelectrical impedance vector analysis (BIVA) have been associated with cell membrane integrity and body cell mass. Zinc is a micronutrient that exerts important structural functions and acts in maintaining cellular functionality. To evaluate cell integrity and body cell mass, PA and BIVA were evaluated in children orally supplemented with zinc at different concentrations. Anthropometric, bioelectrical (resistance and reactance) and serum zinc variables were collected from two randomized, triple-blind, controlled clinical trials. Sampling was composed of 71 children consisting of three groups: a control group who received a placebo and two experimental groups who received oral supplementation of 5 or 10 mg-Zn/day for three months. The three groups presented increases ( Bioelectrical impedance (BIA) has been widely used in clinical practice to evaluate the body composition of adults and children. Compared with other methods for this purpose, the BIA has advantages, including safety, low cost, easy-to-use, portability and practicality . The BIAPA is related to cell integrity and functionality, being important both in healthy people assessment and in prognosis of diseases because it reflects different electrical properties of the body tissues. It also indicates nutritional status and hydration. Its evaluation is superior to other nutritional, anthropometric and serum indicators in different populations ,5,6,7,8.BIVA is useful for clinical purposes because it detects changes in hydration or body composition, as demonstrated by Carrasco-Marginet et al.  and KourThe plasma membrane has three main functions: coating, protection and selective permeability . Zinc is+2 [The participation of zinc in membrane stability is described in the literature through three mechanisms. First, zinc promotes the association between membrane proteins and cytoskeleton proteins. Second, zinc stabilizes the reduced form of sulfhydryl groups, contributing to the antioxidant protection against the effects of membrane rupture caused by lipid and protein oxidation. Third, zinc preserves the integrity of ion channels, thus acting as an antagonist to the adverse effect of free Ca+2 ,16. HoweA database search of two clinical trials, randomized, controlled, triple-blind and non-probabilistic was conducted, had as main objective the supplementation of zinc in apparently healthy children. The partial results of these studies were previously published ,18,19. TThe tests were carried out with different oral zinc supplementations in apparently healthy children , aged between six and nine years. The children were recruited at four municipal schools located in the east and west of the city of Natal, Rio Grande do Norte, Brazil.Exclusion criteria were children in pubarche, thelarche or menarche, who had undergone surgery of any kind, or who used vitamin or mineral supplements. An endocrinologist performed these clinical evaluations. Among the children who comprised this study’s sample, only those eutrophic were included, considering body mass index for age Z-score (BAZ) (Z-scores between −2 and +1) . In addiBoth clinical trials were performed by the same research group and had the same methodology for data collection and analysis with standardized protocols.4·7H2O; Merck, Darmstadt, Germany). Each drop of the supplement contained 1 mg of the zinc element. Zinc sulfate heptahydrate acquisition and the oral zinc solution preparation were performed as described by Brito et al. [Stratification of the sample according to the oral zinc supplementation resulted in the formation of three groups: the control group (CG) received a placebo ; Group 1 (G1) received 5 mg-Zn/day; and Group 2 (G2) received 10 mg-Zn/day. The supplementation period was three months. Zinc was supplied in the form of zinc sulfate heptahydrate . Four milliliters of blood were collected for serum zinc analyses. All processes related to the collection, separation, dilution, and storage of blood for the serum zinc dosage, as well as the calibration of the apparatus, standard serum control, and zinc measurements, were performed as described by Brito et al. . Zinc se2) was calculated as the ratio between body weight and height squared. The weight-for-age (WAZ), height-for-age (HAZ) Z-scores and BAZ were calculated using AnthroPlus software v1.0.4 (available at www.who.int/growthref/en/) and ranked according to the growth curves of the World Health Organization for healthy children aged 5–19 years [The weight and height of the children participating in the trials were measured using an electronic scale  and stadiometer , respectively. BMI  with a single, safe and painless electrical frequency (50 kHz). This tetrapolar method was applied with the subject lying supinated. Four self-adhesive spot electrodes were placed: two on the dorsal surface of the right hand and two on the dorsal surface of the right foot, as described by Lukaski et al. [The bioelectrical impedance parameters, R (Ω) and Xc (Ω), were obtained using the Quantum IIi et al. . With thi et al. , then thR and Xc data were subsequently used to determine PA and BIVA. The PA was calculated with the formula PA = arc tang (Xc/R) × 180/π . The BIVBIVA charts directly measure the vectors R and Xc. According to the RXc chart, children’s standardized impedance measurements are represented as bivariate vectors with their confidence and tolerance intervals, which are ellipses in the RXc plane. These vectors do not depend on equations. To investigate the differences between groups, we plotted the 95% confidence intervals for the mean impedance difference of the bivariate vectors measured under two conditions for each group .The position and length of the vector provide information on the hydration status, cell mass, and cell integrity. It is an upward or downward displacement of the main axis associated with more or less soft tissue cell mass, respectively. A significant value for the T2 statistic is evidence that the mean vectors of each group are different .Statistical analysis was performed by observing the data distribution using the Shapiro–Wilk test. All quantitative variables presented normal distribution and were expressed as mean (standard deviation), except the age that presented non-normal distribution and was expressed as median . Intragroup comparisons were performed using Student’s t-test. p ≤ 0.05).For BIVA, all analyses were performed using the BIVA software . The meaWith the methodology described, data were collected from 71 children aged 6.2–9.9 years. The sex distribution in each group were: 50% both sexes of the CG; and 58% and 39% female in G1 and G2, respectively.p < 0.001) in serum zinc concentrations in all groups, regardless of supplementation  the linear height and weight, but only the group that received a concentration of 10 mg-Zn/day had an improvement in the values of Xc (p = 0.036) and the PA (p = 0.002).p < 0.001) in relation to the confidence ellipses, indicating a possible increase in the cellular mass in these children.Regarding the BIVA , only thOur results show that zinc supplementation modified intrinsic factors related to body composition, such as cell integrity and cell mass, even before changes in serum zinc or anthropometric indicators can be detected, as discussed below.In the population studied, there was an increase in serum zinc over time in all groups, with no differences between them. This study was conducted in a city located on the Atlantic coast, where the supply of food sources rich in zinc is vast. In the study developed by Alves et al. (2016), consumption of energy, protein, fat, carbohydrates, iron and zinc were adequate according to dietary reference intakes by age and sex. Mean zinc intake in these population was 6.00 ± 1.01 mg/ day .Recently, zinc supplementation studies with concentrations ranging from 5 to 50 mg-Zn/day in different infant populations to assess the influence on growth and development have been conducted ,27,28,29The children were in a phase that naturally shows changes in growth. In our study, the anthropometric parameters of WAZ and HAZ did not show differences between the supplemented and control groups. Cho et al. , when evZinc is essential for the integrity and functionality of cell membranes. Its concentration in the cell membrane can be quite high depending on the cell type and is influenced by the nutritional status of zinc in the organism . In the Koury et al.  found hiIn addition to the increase in Xc and PA in the present study, a significant shift to the upper left quadrant was observed in relation to the confidence ellipses in the group that received 10 mg-Zn/day. The use of bioelectric impedance to estimate body composition is a promising methodology . HoweverPiccoli et al.  proposedIn their study, Meleleo et al. (2017) concluded that BIVA could provide more reliable details about differences in body composition in competitive and noncompetitive adolescents . This meA strength of this study was the early detection of changes in bioelectrical parameters as a result of zinc supplementation. Based on that, we suggest that PA and BIVA can be used together to indicate cellular integrity and hydration status, respectively, without requiring predictive equations. These findings encourage the replication of this study in other populations. On the other hand, the sample size and the disregard of other micronutrients, such iron, were limitations of this study. The exclusion of 24 participants with missing data and the distribution of the children in three different groups reduced the sample size and may have affected the power of the study. Iron and calcium interact with zinc and may influence bioelectrical parameters. Thus, the nutrition status of other micronutrients must be considered in future studies with a similar aim.Oral zinc supplementation with 10 mg/day promoted Xc and PA changes, in addition to a vector shift by BIVA, which was associated with changes in cell membrane integrity and an increase in cell mass in eutrophic children. These original results were observed before changes in serum zinc concentrations or anthropometric indicators.


A two-step cluster analysis classified three groups according to sedentary PA and MVPA. The Sedentary (56.9%), presented the highest sedentary PA ; The Hazardous (19.4%) had the lowest values of sedentary and MVPA . Balanced (23.7%) had the highest MVPA (28.61 [27.16–30.07] min/hour). Sedentary had the lowest count of home time associated (20%) and higher school (26%) time when compared to the Hazardous (13%). The Balanced showed the highest count of school (61%) and home time (47%). Different profiles for young athletes revealed alarming behavior in the associations with sedentary PA, sitting and SED breaks, which may influence performance and health.The level of physical activity (PA) and sedentary behavior (SED) off-training of young athletes may reveal the quality of recovery from training and highlight health related issues. Thus, the aim was to identify and describe young athletes’ PA and SED off-training, according to daily life activities. Eight athletes  of a sport talent program wore on their waist a tri-axial accelerometer (ActiGraph Physical activity (PA) and sedentary behavior (SED) profiles of healthy ,2,3 and Previous studies demonstrated independency between time spent in MVPA and SED by evidencing that people who accomplish the recommended MVPA levels, when compared to sedentary ones, do not present less SED time ,11. HoweTo young athletes, in the perspective of sports performance, PA and SED profiles should also be considered key to evaluating, preventing and treating overtraining symptoms . Indeed,Thus far, two studies have analyzed the PA and SED profiles during off-training hours and it seems that elite athletes have higher levels of SED during waking hours when compared to non-athletes ,18. ThisThe efforts for measuring, interpreting and managing internal and external training loads in sports  or even In line with that, wearable accelerometers have been extensively used as a non-invasive way to estimate energy expenditure levels in both competitive and free-living environments ,29,30,31Eight young athletes  of a high-level sports talent program were recruited to participate in this study. Criteria for inclusion were applied to ensure all athletes were aged 13–17 years, were engaged in a minimum of six hours of training per week , and had professional experience of no less than five years of regular participation at previous competitive seasons. Athletes, their legal guardians, and coaches were fully informed about the purpose, benefits and risks of the study, and provided written informed consent before the study started. The study protocol conformed to the recommendations of the Declaration of Helsinki. It was approved and followed the guidelines stated by the local Institutional Research Ethics Committee.® GT9X Link +  to monitor their PA and SED, as well as the instructions of how to use it. They should wear the monitor in the dominant side of their hip, for 15 consecutive days, completing valid mean wear time of 10.6 ± 1.0 h per day. They were asked to not use the devices during training periods, bed time, personal care and activities under water. Participants were asked to fill a daily report with a rough description of the periods of monitor wearing and non-wearing. The participants also described in this report their schedule of activities performed during the day, according to the following categories: school, home time, leisure time, training, and competition. The second evaluation moment was arranged at the end of accelerometer data recording period to collect the monitors and the daily report.The protocol design consisted of two evaluation moments. In the first moment, the athletes received the Actigraph® 6.10 software . The data were chunked in 60-s epochs. Sixty or more minutes of zero accelerometer counts were considered non-wear time, therefore excluded from analysis. Valid accelerometer wear time consisted of, at least, 600 min of accelerometer wear time per day [−1, light PA was 181–756 counts·min−1, moderate PA was 757–1111 counts·min−1, and vigorous PA was ≥1112 counts·min−1. Sedentary breaks (SED breaks) were as defined as time spent in a minimum of 30 min of prolonged sedentary activity specifically in each waking hour of the day. The sedentary behavior was analyzed through the ActiGraph® inclinometer measures and associated to the following modes: sitting, laying and standing.As a valid instrument to measure PA and SED in free-living environment , the tri per day  and a mi per day . The cut per day : sedentaAll PA and SED variables were analyzed in minutes per hour during off-training time. The athletes were classified in groups using a two-step cluster analysis according to the hourly distribution of MVPA and sedentary PA, which was considered the main variables in representing PA and SED off-training beforehand. One-way ANOVA was used to test the differences between MVPA and sedentary PA among the groups defined by the cluster model.® , with p < 0.05.Then, how other PA and SED variables  would distinguish the clustered groups was tested, as well as its efficiency in correctly classifying the original groups, but only for the periods where SED breaks were higher than 30 min, using discriminant function analysis. The effect sizes of the discriminant functions were assessed by Wilks’ lambda and squared canonical correlation, which explains the variance associated with each function . AdditioThis study aimed to identify and describe young athletes’ level of PA and SED off-training and according to their usual activities schedule. Thus, the two-step cluster analysis revealed that sedentary PA and total MVPA can be grouped into three different clusters, with a good average silhouette coefficient of 0.6 . Means ap < 0.001). Vigorous PA, steps counts, energy expenditure, standing, moderate PA and MET had the highest correlation with the first function, while lying, SED breaks, sitting and light PA had highest correlation to the second function (The discriminant analysis revealed that two functions can explain all the variance contained in the dataset. The first and second functions hold 70.4% and 29.6% of the total variance explained, respectively, with statistically significant Wilks’ lambda (0.21 and 0.58 for Functions 1 and 2, respectively, function . The scoThe present study characterized the PA and SED of young athletes during non-training hours. Findings revealed three distinct profiles of MVPA and sedentary PA spent on daily basis. The Sedentary group showed the highest mean sedentary PA per hour of waking time, while the Balanced group presented the highest mean MVPA per hour of waking time, and the Hazardous group had the lowest sedentary PA and MVPA. Both Sedentary and Balanced, however, achieve the WHO daily recommended levels of MVPA, considering the mean wear time for the sample of this study was 11 h. Previous studies have reported that PA and SED are distinct domains not only for those who do not meet the recommended guideline levels, but also for those who do, including young people  and athlThus, present results indicate that the displacement among these behaviors is also present in young athletes’ lifestyle. Indeed, the levels of sedentary PA off-training found for all groups has already been reported for elite professional soccer players at post-training time as well . Non-traThe Balanced group was characterized by higher MVPA, light, moderate and vigorous PA, as well as step counts. The first discriminant function, which explains most of the data variability, shows high and significant correlations to these variables. This group did not present the lowest daily time in sedentary PA; hence, off-training, it seems that the athletes classified in this group better manage their PA and SED profiles throughout the day. The Balanced group showed the highest count percentages of school and home time, indicating that their profiles are not affected by the main environments attended in daily life, thus there is viability in finding balance between PA and SED off-training in modern daily life for athletes. This could be used as parameter of reference for outcomes in the planning of programs and activities for the ones classified in the other groups.This is not the case for the Hazardous group, which showed the lowest mean daily time spent in all PA and SED variables. This result did not avoid the number of SED breaks, however, being the highest among the groups. Prolonged sedentariness in children and adolescents is associated with increased body fat in athletes ,11, obesIt might be argued that young elite athletes being exposed to hours of training and competitions is enough to address this issue while enhancing fitness components  and decrAge- and gender-related patterns influence time use in terms of PA and SED , but parThe aim of this study was to identify and describe young athletes’ level of PA and SED off-training and according to their usual activities schedule in a typical week. Different patterns were found for young athletes’ PA and SED profiles, revealing occurrence of alarming behavior in the time spent in sedentary PA, sitting and prolonged sedentary breaks during the day. Most weekdays waking hours are spent at school or home, which are places that promote sedentarism. However, some athletes still manage to balance healthy PA and SED levels. The awareness of such profiles is important for performance, which might help in the recovering process of training, as well as for health, once it decreases the harmful influences of the inevitable modern sedentarism. Thus, this information may provide useful data and insights for helping coaches and trainers in controlling the loads that may influence training performance as well as health status.


Virgula Marls and Banné Marls) in the area of Porrentruy . This material was revealed by construction works of the A16 Transjurane highway between 2000 and 2011, and led to the recent description of the new species Plesiochelys bigleri. In the years 2014 and 2016, new fragmentary turtle material was collected from the Banné Marls  near the village of Glovelier, Canton of Jura, Switzerland. The new material consists of a complete shell, additional shell elements, a few bones from the appendicular and vertebral skeleton, and a fragmentary basicranium. This material can be confidently assigned to the species P. bigleri. It supports the presence of this species in the Banné Marls, slightly extends its spatial distribution and confirms the differences with the closely related species P. etalloni. The new material reveals that the split between the cerebral and palatine branches of the internal carotid artery occurs in a vertical plane in P. bigleri. This condition could not be observed in the type material due to poor preservation. This new character clearly distinguishes P. bigleri from P. etalloni and seems to be unique among thalassochelydians.Plesiochelyidae were relatively large coastal marine turtles, which inhabited the epicontinental seas of Western Europe during the Late Jurassic. Their fossil record can be tracked in Germany, Switzerland, the United Kingdom, France, Spain and Portugal. The Jura Mountains, in northwestern Switzerland, have been the main source for the study of this group, mostly thanks to the rich and famous historical locality of Solothurn. In the last two decades, numerous plesiochelyid remains have been collected from Kimmeridgian deposits (Lower  Virgula Marls of the upper Kimmeridgian and the Banné Marls of the lower Kimmeridgian ..1].Referred material: MJSN CBE-0001, shell fragments including articulated and isolated costals, a peripheral, and a partial scapula; MJSN CBE-0002, a partial basicranium with the left otic chamber and columella auris; MJSN CBE-0003, a near complete shell with the left femur, right pubis, a cervical vertebra, and a caudal vertebra.Plesiochelys bigleri was recently described in detail )etalloni . In all Plesiochelys bigleri), and NMS 40870  were CT-scanned and segmented for the purpose of this study  reveals several morphological features and bone contacts that were not very clear or could not at all be observed in the type material of Plesiochelys bigleri. Especially noteworthy, is the spatial orientation of the split between the cerebral and palatine branches of the carotid artery. In thalassochelydians, this split occurs in a horizontal plane, while in this specimen, the split is oriented on a vertical plane instead. This condition seems to be unique for this clade. MJSN CBE-0003 is the first specimen to document the 3D shape of the shell of Plesiochelys bigleri. This rather low shell shows two parasagittal bulges on the anterior costals that may be considered as a new character of P. bigleri.The newly obtained material from Glovelier can be confidently referred to S1 FigViews: dorsal (A), ventral (B), anterior (C), posterior (D), medial (E), lateral (F).(TIF)Click here for additional data file.S2 FigViews: A: dorsal, B: ventral, C: anterior, D: posterior, E: medial, F: lateral.(TIF)Click here for additional data file.


This study aimed to evaluate the occurrence of gluten contamination in naturally gluten-free meals from food services in the Federal District, Brazil. This is an exploratory cross-sectional quantitative study in which a total of 180 samples of naturally gluten-free dishes were collected from 60 food services in Brazil. The enzyme-linked immunosorbent assay was used for the quantification of gluten. As established by the Codex Alimentarius, the threshold of 20 ppm of gluten was considered as the accepted upper gluten level for gluten-free food. A total of 2.8% (95% CI: 0.3–5.2%) gluten contamination was found in the samples. Among the 60 food services, 6.7% (95% CI: 2.7–10.6%) displayed at least one contaminated food in our sample. The occurrence of gluten contamination in naturally gluten-free preparations was uncommon and low on a quantitative basis. Celiac disease (CD) is an immune-mediated enteropathy disorder triggered by the consumption of gluten in genetically predisposed individuals, which affects about 1% of the worldwide population . StandarBesides CD, there are other health conditions related to gluten that require a gluten-free diet (GFD). The gluten-related disorders (GRD) comprise three primary forms of gluten reactions: allergic , autoimmune , and possibly immune-mediated (gluten sensitivity) . These cIn CD individuals, adherence to the GFD results in a remission of symptoms, normalization of serum autoantibodies typical of the disease, and recovery of intestinal histological architecture . HoweverThe presence of gluten traces in food may be enough to cause health problems in CD. In a systematic review, authors found that some patients tolerate an average of 34–36 mg of gluten per day. However, others develop mucosal abnormalities while consuming 10 mg of gluten per day. Therefore, the authors mention that although there is no evidence to recommend a single definitive threshold, a daily gluten intake of less than 10 mg is unlikely to provoke significant histological alterations [As proposed by the Codex Alimentarius, in the gluten-free foods (GFF), the gluten level cannot exceed 20 ppm (mg/kg) in total . ConsideHence, eating products from the food industry or food from restaurants may represent a risk for CD patients ,14. The Some studies have investigated the occurrence of gluten contamination in processed food. In Italy, the authors found a 9% rate of contamination in gluten-free products available in the Italian market . In BrazBesides preventing histological alterations and clinical manifestations of CD, the GFD is crucial for reducing the risk for long-term complications . HoweverThe recognition of CD as a public health problem has increased nowadays  and, in Mesa Brasil, which is a national network of food banks that aim to combat hunger and food waste, was also used. At last, a list of the government community restaurants was provided by the Subsecretariat for Food and Nutrition Security of the state.According to the Brazilian resolution n° 216 from the National Health Surveillance Agency, “food services” include establishments such as canteens, buffets, confectioners, industrial kitchens, institutional kitchens, delicatessens, snack bars, bakeries, pastry shops, restaurants, rotisseries, and similar items . For theA total of 282 establishments were obtained considering these three databases. These food services comprise restaurants where Brazilians commonly have lunch, including commercial restaurants, governmental restaurants, and institutional restaurants . Those are regular establishments that produce both gluten-containing food, such as pasta and pies, for example, and naturally GFF, such as rice, meat, beans, and salads, among other items.Food services were invited to participate in the study by telephone and/or email contact, and a total of 60 establishments agreed to take part. This sample size allowed estimating a 95% confidence interval of the prevalence of gluten contamination with an error of estimation of 4%.It is important to clarify that control of gluten cross-contamination in restaurants is not a regular practice in Brazil. The Brazilian legislation does not require cross-contamination prevention in food services. Furthermore, it does not set a limit of gluten content for food considered gluten-free. In Brazil, all food that is naturally gluten-free or produced with gluten-free ingredients can be identified as “gluten-free” without the need for laboratory analysis. Therefore, refusal to participate in the study was not believed to be related to fear of identification of contaminated samples in the establishment, since this is not a concern of nutritionists and owners of food services in general. Common reasons to deny participation in the study were overloaded routine and lack of available time to accommodate the researchers.The Ethics Committee of the Faculty of Health Sciences of University of Brasilia approved the study protocol (protocol number: 60987816000000030). The person in charge of each food service signed an agreement term for authorization of the study in the establishment.Food services were visited during the usual production period of lunch between January and November 2017. The day menu was requested to the chef or nutritionist responsible for the establishment. Since the labeling of gluten-free foods in restaurants is not required by legislation in Brazil, the majority of food services did not classify menu dishes in regards to gluten content. Researchers evaluated the menu in each establishment on the day of the visit and identified traditional and inherently gluten-free dishes . Moreover, the label of all the packaged ingredients used to prepare those dishes was checked . Foods selected for collection were chosen randomly among those naturally gluten-free dishes. A total of three different samples (different food items) were collected in each food service.Since the study aimed to detect accidental gluten contamination in naturally GFF, all the ingredients used in the food samples selected were checked by verification of the label in order to ensure that no intentional gluten-containing ingredient was used in the food collected. In case any gluten-containing ingredient was used in a naturally gluten-free dish—for example, wheat flour to thicken a meat sauce—this dish was not considered for the collection process, since it would obviously present gluten in its composition.In all the establishments, gluten-free and gluten-containing food were produced in the same production area, as checked by the researcher. Three food samples were collected in each establishment, generating a total of 180 samples, categorized according to the type of food in: (i) main course (meats and vegetarian protein options—52 samples); (ii) garnish ; (iii) side dishes ; and (iv) others (two samples).Foods were categorized into three types because the lunch meal in Brazil is traditionally composed mainly by these dishes . In cases where there was no gluten-free food alternative from the above categories, another gluten-free dish of the day was also randomly selected (category “others”) such as salads (one sample) and desserts (one sample). Bakery/bread/pizza and dairy are not usually consumed at lunch in Brazil. Although desserts may be present in the lunch meal, not all establishments serve dessert on a daily basis.Handling of samples was conducted at the Laboratório de Técnica Dietética, University of Brasilia, Brazil. Before processing the samples, laboratory surfaces, instruments, and equipment were cleaned with water, regular detergent, Triton X 5% solution (a detergent used to remove proteins such as gluten), distilled water, and ethanol solution (70%) in order to prevent any gluten contamination from the environment. This procedure was also performed between the homogenization processes of each sample, to prevent contamination among them, as proposed by Silva . SamplesGluten content of samples was determined by the Ridascreen Gliadin sandwich R5 enzyme-linked immunosorbent assay R-7001  at the Laboratório Interdisciplinar de Biociências (Laboratório de Pesquisas em Doença Celíaca), University of Brasília, Brazil. The manufacturer’s instructions were strictly followed, as described below.For the extraction of gluten from the food matrix, the extraction cocktail solution (R7016) from R-Biopharm was used . One test portion of 0.25 g of each homogenized sample was weighed and mixed with 2.5 mL of the cocktail in pre-labeled falcon tubes. Samples were incubated for 40 min at 50 °C (122 °F). After being cooled down, samples were mixed with 7.5 mL of 80% ethanol and then shaken by a rotator at room temperature for 1 h. Then, samples were centrifuged at 2500 g for 10 min, and the supernatants were transferred to other falcon tubes and stored at room temperature in the dark until the date of the ELISA step, which was performed in one to two days after the extraction step, respecting the 8-week sample viability period.®  automated microplate immunoanalyzer. An amount of 100 μL of each standard solution, samples, and controls were added to separate triplicate wells and incubated for 30 min at room temperature. Three washing steps were performed using the washing buffer. An amount of 100 μL of the diluted enzyme conjugate was added to each well, followed by an incubation of 30 min at room temperature, and again three washing steps. Then, 50 μL of the substrate and 50 μL of chromogen were added to each well, followed by an incubation period of 30 min at room temperature in the dark. At last, 100 μL of the stop reagent was added to each well, and the absorbance was measured at 450 nm within 30 min after the addition of the stop solution.For the ELISA step, extracted samples were diluted with diluted sample diluent at 1:12.5. All of the next steps were automatically performed by the Best 2000Results were calculated using a cubic spline function, as suggested by the manufacturer of the ELISA kit. The lower limit of quantification of the kit is 2.5 ppm (mg/kg) of gliadin, corresponding to 5 ppm (mg/kg) of gluten. As established by the Codex Alimentarius, the threshold of 20 ppm of gluten was considered as the accepted upper gluten level for GFF . Foods cFor quality control of the extraction step, the test implementation, and the handling of the Ridascreen Gliadin (R7001), the processed Gliadin Assay Controls (R7012) from R-Biopharm were used.For data analysis, the GraphPad Prism, version 6 , and the Microsoft Excel, version 2013 , software were used. The number of meals served daily at lunch were presented as mean ± standard deviation (SD). The confidence intervals of the prevalence of gluten contamination in food samples were obtained by the normal approximation. In the food services, the confidence intervals also considered the correction for finite populations.A total of 60 food services agreed to participate in this study. The characteristics of the food services are summarized in Institutional restaurants include the food services of government agencies, refectories of philanthropic institutions, and refectories for employees at industries, hospitals, and hotels in the city. These establishments accounted for 30% of the sample sites. It is noteworthy that the number of meals served daily at lunch is high (1076 ± 1807) at these food services, since a large portion of the population does not return home for this meal and consequently eats at the workplace.Daycare centers and school canteens primarily serve children and adolescents, but also serve the local staff. These establishments accounted for 30% of the sample. The number of meals served daily at lunch is 234 ± 148.Community restaurants represented 23.3% of the sample sites. These establishments are part of a government assistance program aimed at guaranteeing access to food for the low-income population ,28. CommCommercial restaurants accounted for 16.7% of the study sample. These establishments are distributed throughout the city, and customers who attend these locations have different profiles, varying from children to the elderly. These food services have a self-service system in which the customers pick as much as they want from a wide range of dishes. The customer’s plate is weighed and the price paid for the meal is proportional to the amount of food. This type of establishment is common in Brazil. The number of meals served daily at lunch is 233 ± 105.The distribution of gluten concentration of the collected samples are shown in Among the contaminated samples, two were from the same establishment. Therefore, out of 60 food services, four displayed contaminated foods . The contaminated samples were: braised pumpkin and okra (garnish); roast beef, grilled steak, and Provençal chicken (main course); and black bean (side dish). These samples came from the following types of food services: an industry refectory; a school canteen; a commercial restaurant; and a community restaurant (two contaminated samples). When analyzing contamination by type of food, the main course was the category with the highest percentage of contamination among its samples, though it was still relatively low (5.8%), followed by garnish (1.8% contamination) and side dishes (1.4% contamination). Among the five contaminated samples, four displayed gluten concentration above the limit of quantification of the ELISA test (80 ppm), and the other sample presented 22.8 ppm of gluten.Most of the foods Brazilians eat at lunch are naturally GFF, such as rice, beans, salad, meats, and vegetables . TherefoAlthough restaurants from our sample have nutritionists who are technically responsible for the establishment and quality of the meals served, this did not guarantee the absence of gluten in all the foods, which indicates the need for supervision and better guidance of employees in regards to food preparation procedures.It is important to emphasize that gluten-contamination control is not a regular practice in food services from Brazil. The Brazilian legislation does not require cross-contamination prevention in meals from restaurants . A studyOur sample was composed of different types of food services that attend to diverse publics. Thus, these establishments cover various possibilities of lunch options for celiac individuals, contemplating profiles of different age groups and socio-economic conditions, among other aspects.Gluten contamination above 20 ppm was present in 2.8% of the foods sampled. The occurrence of gluten contamination is lower than the one found by Oliveira et al.  in 60 beIn another study from Brazil, results revealed an occurrence of 21.5% of contamination in bakery products. Nevertheless, a higher percentage of contamination is expected in this type of establishment, since there is an extensive manipulation of wheat flour for the production of bread, cakes, biscuits, and similar, which increases chances of cross-contamination .In regard to the type of food analyzed, out of the five contaminated food samples, three were main courses. One of these samples—Provençal chicken—is served with sauce. In the study by Simpson and Thompson , the autThe correct information about the presence/absence of gluten in food provided by food services is very important for those individuals who need to maintain a GFD. In a systematic review, 24 cross-sectional studies that assessed gluten-contamination in gluten-free industrial and non-industrial products were evaluated. Results from the statistical meta-analysis pointed out a contamination prevalence of 13.2% (95% CI: 10.8–15.7%) in industrial products and 41.5% (95% CI: 16.6–66.4%) in non-industrial food products. Industrial products labeled as “gluten-free” showed a lower percentage of gluten contamination in comparison to non-industrialized products. Despite that, any contaminated sample found in labeled “gluten-free” industrial food presents greater relevance, since it indicates that the label may be unreliable, and these “gluten-free” products should not be considered safe for celiac patients . In BrazAs advocated by the Codex Alimentarius, GFF must be prepared with special care under Good Manufacturing Practices (GMP) in order to avoid gluten cross-contamination . In the In Brazil, a checklist specifically elaborated for the prevention of gluten cross-contamination in food services has been created, and it is currently undergoing a process of validation .Data collected in the study by Petruzzelli et al.  also revIn the present study, gluten-containing and gluten-free meals were produced in the same facility area, without separation of routines, utensils, and equipment. Even so, the occurrence of gluten contamination was not as high as expected. As a matter of fact, it was quite low. The majority of the samples displayed gluten content below the quantification limit. It is important to mention that there is a big concern regarding gluten cross-contamination in the nutritional assessment of CD patients. These individuals are regularly advised on this subject, and it is a topic often discussed in the scientific literature ,35,40.Nevertheless, it is noteworthy that staff in food services is usually not well educated in regards to the subject of CD and gluten ,31,33. AAlthough the treatment for CD could be considered relatively simple from a medical point of view since it does not require regular use of medication and/or medical intervention procedures, it brings a lot of social compromise to the patient’s life ,42,43,44The need to avoid gluten-containing food often brings considerable changes in eating patterns and lifestyle that may be difficult to maintain over a lifetime . Thus, iFuture research should focus on identifying categories of foods and types of food services that are more susceptible to the occurrence of cross-contamination and what inadequate procedures and practices indeed lead to a contaminated final product or meal.Furthermore, long working hours typical of the routine of many individuals nowadays make it hard for people to return home for lunch, which leads to an increasing trend of having meals in establishments such as self-service and fast-food restaurants . TherefoThe highlights of this study include the sample being composed of a great diversity of types of food services that may be frequented by CD patients, which enriches the data obtained. Moreover, there are very few data in the literature regarding gluten cross-contamination in meals from food services. However, it is important to mention the limitations of the research, such as the size of the sample of restaurants (n = 60) due to the low response rate among contacted food services to participate in the study, and also the difficulties regarding the comparison of results due to the lack of similar studies. In addition to the number of restaurants, depending on the homogenization and distribution of gluten after sample preparation, the sample size (0.25 g) used in ELISA may not be enough to measure gluten in the meal, especially when only one test portion was analyzed. This limits the assessment of variability between gluten measurements within a sample. Moreover, only three food samples from each site were collected. It is possible that other foods that were not analyzed in this study had gluten content above the allowed limit.Another limitation refers to the analytical method used. Although it is accepted as the gold-standard for gluten analysis by the Codex Alimentarius and the Association of Official Analytical Chemists (AOAC), the ELISA R5 technique requires at least two epitopes in the protein. Nevertheless, proteins are fragmented during food processing and may be converted into peptides in which only one toxic epitope appears, which might compromise part of the results .Although there is much concern regarding the presence of gluten traces in supposedly GFF, the occurrence of contamination found in meals from food services in this study was low. In a certain way, this creates the prospect of reliability for celiac patients when eating out-of-home in Brazil. However, this issue must be carefully assessed.Adherence to the GFD is essential for the clinical, histological, and serological recovery of patients with CD and consequent improvement in the quality of life. Despite the primarily dietary treatment, people with CD face several difficulties. In this context, the fear of being involuntarily exposed to gluten through the consumption of contaminated food stands out.Despite the findings from this study, an effort should be made by food services and also the food industry in order to minimize the risk of cross-contamination in food. That would create a more reliable scenario for CD patients who need to eat out-of-home. Furthermore, future research should focus on identifying inappropriate procedures that cause gluten-contamination and proposing new strategies to overcome this issue.


The causes of childhood cancer are not well known, but the advanced age of the parents has been suggested as a risk factor for childhood cancer in several observational studies. In this study, we examine a possible link between parental age and childhood solid tumors. We conducted a hospital-based case-control study  at Rebagliati Hospital, Lima, Peru. Odd ratio was used to compare categories of advancing maternal and paternal age with and without adjusting for possible confounding factors were calculated. The risk of childhood retinoblastoma was significantly higher among children of mothers aged> 35 years  and fathers aged> 35 years . A significant trend with increasing mother's age (p = 0.037) and father's age (p = 0.005) was found. There were more risks to development of non-Hodgkin's lymphoma (p = 0.047) and gonadal germ cell tumors (p = 0.04) for advanced paternal age. There was a strong protective effect of increasing parity on risk of solid tumors in children (p=0.0015). Our results suggest that advanced parental age is associated with the risk for the development of retinoblastoma. Advanced paternal age increases the risk of non-Hodgkin lymphoma and gonadal germ cell tumor. The higher the order of birth of the children, the less the chance of developing any neoplasm. Although the childhood cancer is a rare disease, during the last years, appearance of children's cancer had a higher incidence rate, affecting approximately one in 435 children under age 15 years –4), and Family structures have changed in relation to past generations since nowadays, when maternity/paternity occurs, advancing maternal or paternal age and a lower number of children are also more likely to occur. Additionally, advanced maternal age has been positively linked with higher risk of having a child with Down syndrome, among other congenital disorders, which in turn has a higher incidence of acute myeloid and lymphoid leukaemia . AdvanceThe link between advanced maternal and paternal age and a higher incidence of the appearance of children's cancer has been revealed in studies conducted among patients with leukaemia –9, lymphTo date, it has not been made clear if advanced parental age is linked with higher risk of children's cancer; hence, the present study aims to determine whether advancing parental age is associated with an increased risk of childhood cancer in offspring.A case-control study based on hospital records  was conducted. Both, cases and controls, included children and adolescents younger than 18. For every case, 2 age-and sex-comparable controls were assigned .Cases had an anatomical pathological diagnosis of Hodgkin and Non-Hodgkin lymphoma, brain tumors, germ cell tumors, and any solid tumors, between the years 2012 and 2015. Controls were children who were hospitalized for nononcological diseases  during the same period.Data collection was conducted via questionnaires to parents about their medical histories and their families' medical histories, together with a revision of every patient or control patient's medical record.The data was analysed using conditional logistic regression for studies involving the cases and controls. Relative risks were estimated by odd ratios (OR) with a 95% confidence interval (CI) with a primary analysis between the link between maternal and paternal age as categorical variable according to age group  and as quantitative variable and childhood cancer diagnosis. We established raw OR and OR adjusted to confounding variables; these were chosen based on their prior observed association with childhood cancers –14: pateThis study has been evaluated and approved by the Ethics Commission of our institution prior to project execution. All clinical and sociodemographic data comply with confidentiality norms to protect the identity of patients.Between 2012 and 2015, 310 cases of childhood cancer (solid tumors and lymphomas) were identified; we compared those cases with 620 controls, matched according to age (with the margin of six months), sex, and geographical area of origin .The presence of childhood cancer was slightly higher among males. Parental age was grouped into a 5-year group. The main age of case mothers at birth was slightly older (28.92 [SD=6.30]) than control mothers (28.47[6.16]); contrary pattern was observed with the main age of fathers, slightly younger in cases (31.74 [6.82]) than controls (32.37 [7.82]). The only statistically significant difference between controls and cases was in the order of birth, since the difference observed in the level of education of the father is influence by the number of lost data.In our case-control study, maternal age shows positive linear trends (age groups are taken to be continuous) for 2 childhood cancer groups: osteosarcoma (p=0.019) and retinoblastoma (p=0.04); however after adjustment for covariates , maternal age only indicated a positive linear trend for retinoblastoma (p=0.037). In addition, the risk of development retinoblastoma was significantly higher among children of mothers older than 35 years old ; such association did not occur in other neoplasms. .Similarly, paternal age demonstrated positive linear trends (age groups are taken to be continuous) for 2 childhood cancer groups: gonadal germinal tumor (p=0.03) and retinoblastoma (p=0.03). After adjustment for covariates , paternal age continued showing a positive linear trend for non-Hodgkin lymphoma (p=0.047), gonadal germinal tumor (p=0.04), and retinoblastoma (p=0.037). Additionally, the risk of development retinoblastoma was significantly higher among children of fathers older than 35 years old . .Furthermore, a protective effect against risk of solid tumors when parity increased (p=0.0015) was found. It was also observed that there was a significant decrease in the risk of children's neoplasms as the number of siblings in a family increased; there is a protective effect from the third child . We decided to do the analysis of all the neoplasias together, due to having few cases to do a separated analysis. .This work represents the first published study focused on advanced parental age  as a risk factor associated with the appearance of solid tumors in children in Peru.Advanced parental age, considered in some studies as older than 35 or 40 years old, has been frequently linked with higher risk of children's malignant neoplasms –13, withIn our case-control study, we observed a slight rise in the risk of developing retinoblastoma by children of advanced-age mothers and fathers. For this cancer, the oldest maternal age group (>35 years) showed an elevated risk with adjusted OR de 1.21 (1.09-6-08) compared with other exposure age groups.These results are similar to other studies. One of the most comprehensive cohort studies done by Yip et al.  describeHowever, the study of Johnson et al. (2009)  did not In the case of the effect of father's age, our study showed positive linear trend for the risk of retinoblastoma after adjusted by maternal age (p=0.005) and showed an increased risk in those parents who were older than 35 . These results were different to those found by Yip et al.  that didIn spite of those findings, general analysis of published studies does not make it clear that there is a fundamental difference in favour of advanced age of parents as the cause of increased risk of retinoblastoma among children.In our study, apart from the positive association between advanced maternal age and risk of retinoblastoma, we did not find a relationship between advanced maternal and risk for other solid tumors, like as brain tumors, Wilms tumors, Ewing's sarcoma, soft tissue sarcoma, hepatoblastoma, Hodgkin lymphoma, and neuroblastoma. This is different to the report of Johnson et al. , who fouInterestingly, our study showed that single children had a higher risk of developing childhood cancer than children who had siblings. That is, the greater the order of birth of a child, the lower the risk of cancer (p=0.0015), and we could see a protective effect from the third child , which continued for the fourth  and fifth child . These results were similar to those reported by Von Behren et al. (2011) , who fouDue to its observational nature, this study presents strengths and weaknesses. One of its main strengths is that it is the first study of its kind conducted in our country and region. Its main weakness lies in the number of cases included in the study, because these pathologies represent diseases, which are relatively infrequent. Further limitations also include employing hospital-type controls and missing data in some variables; therefore, the results obtained in this study cannot be generalized to the Peruvian population and are only useful in the hospital setting in which the study was carried out. Nowadays in Peru, it is virtually impossible to apply population studies into children's cancer because Peruvian hospitals do not have registration systems of optimal quality.Our study suggests that advanced parental  age, ≥ 35 years old, increases the risk of developing retinoblastoma in children; at advanced paternal age increases the risk of developing non-Hodgkin lymphoma and gonadal germ cell tumors; and finally, this study observed that the higher the order of birth of the children, the less chance of developing any neoplasm. Additional studies with larger population-based samples are needed to confirm our hypothesis.


M = 1.20, SE = 0.06, CI = 1.08 to 1.32) had significantly higher daily PA than the HIV+ group . The HIV- group  also reported participating significantly more in muscle strengthening activities than the HIV+ group . Multiple regression analyses showed that higher PASE (p < .001) and greater enjoyment of PA (p < .01) were predictive of higher daily PA. HIV- participants had higher PASE but lower BWS compared to HIV+ participants. Sex and age differences were observed in muscle strengthening activities and sedentary behavior. This study supports previous findings on the association of efficacy beliefs to daily PA and muscle strengthening activities. The findings have implications for PA interventions aimed at health promotion and mitigation of the effects of living with HIV/AIDS.The objectives of this study were to examine differences in physical activity behaviors as a function of human immunodeficiency virus (HIV) status and sex, to test differences in physical activity self-efficacy (PASE), body weight satisfaction (BWS), and enjoyment of physical activity as a function of HIV status, and to determine if PASE, BWS, and enjoyment are associated with daily physical activity (daily PA), muscle strengthening activities, and sedentary behavior of youth with and without HIV. A total of 250 HIV positive (HIV+) and HIV negative (HIV-) youth from Botswana aged 12–23 years  participated in the study. The HIV+ group (n = 88) was recruited from a previous 12-month antiretroviral therapy (ART) and nutrition intervention study. The HIV- group (n = 162) was randomly selected from public junior and senior (secondary) high schools in and around Gaborone. Participants’ PASE, BWS, enjoyment of physical activity, daily PA, muscle strengthening, body mass index (BMI), and sedentary behavior were obtained using items from the Youth Risk Behavior Surveillance Survey. Multivariate analysis of variance (MANOVA) showed that the HIV- group ( Self-efficacy is one of the most studied psychological correlates of physical activity (PA) and exercise behaviors –3. A keyEnjoyment of PA relates to intrinsic motives, such as moods, feelings, and attitudes, including the sheer pleasure of participation in PA . EnjoymeBWS refers to the discrepancy between one’s current/actual and ideal body image and body weight with scores of a lower magnitude suggesting satisfaction and scores of a higher magnitude suggesting dissatisfaction . BWS is Cultural perceptions of body image and body weight and their potential impact on enjoyment of PA and BWS cannot be ignored. Some studies suggest African societies are likely to associate thinness with ill-health and lower socio-economic status, while thinness might be viewed positively in most western societies ,27. ThisHowever, there is limited data on PA and sedentary behavior for children and youth living with HIV and on ART, specially in developing countries. This is despite the evidence that shows that individuals living with HIV are susceptible to debilitating psychological symptoms including chronic fatigue and anxiety , which cTo provide more context, the most recent global PA data among youth indicates that PA levels are generally below the evidence-based guidelines of 60 minutes of daily moderate-vigorous intensity activity ,13. HoweIn one large-scale study of 34 African countries, researchers examined PA and sedentary behaviors among adolescents aged 13–15 years old . The resLimited data on the relationships among PASE, BWS, enjoyment of PA, and PA and sedentary behavior among youth in an African context makes it imperative to investigate these relationships. Given the existing evidence of a stigma that associates thinness with ill-health in Southern African populations, it will be interesting to examine variations in BWS and PA among HIV+ and HIV- youth. Such evidence is likely to offer a better understanding of how efficacy beliefs and BWS may affect or be affected by participation in PA among individuals living with HIV. The evidence may also be useful to the development of suitable strategies for the promotion of PA among people living HIV. Therefore, the purposes of this study were to:Determine if differences exist in daily PA, muscle strengthening activities, and sedentary behavior by HIV status and sex, controlling for age and BMI.Determine if psychosocial correlates of PA  differ as a function of HIV status, controlling for age, sex, and BMI.Examine if PASE, BWS, and enjoyment of PA are associated with daily PA, muscle strengthening activities, and sedentary behavior, controlling for age, sex, and BMI.A total of 250 HIV+ and HIV- youth  aged 12–23 years  participated in the study. The HIV+ group (n = 88) was part of a previous 12-month ART and nutrition randomized control trial involving 201 children aged 5–12 years old ,35. PartAfter all the required ethical clearance had been obtained, the HIV+ group was contacted by phone and invited to participate in the study. The invitations were done through the pediatric clinic which had their contact information. A total of 108 out of the possible 201 participants responded to the phone calls and, of these, 88 agreed to take part in the current study. Twenty declined to participate because of constraints such as living in another town that was too far away from the study center. Ninety-two did not return the calls or the calls did not go through. The research team visited target schools to recruit participants to the HIV- group. Information about the study was passed through school counsellors. After participants and their parents gave consent and assent to take part in the study, as well as share the results from the HIV tests with the study, they were referred to the voluntary counselling and testing center for HIV screening. The HIV counselling and testing centers are manned by personnel who have been trained on HIV-related counselling and testing. Out of a total of 169 students recruited and referred for screening, seven declined to take part in the study, resulting in a 96% response rate. Study assessments were done on weekends at a youth facility in Gaborone. All participants completed questionnaires on their own, with the help of trained research assistants where necessary. The questionnaires were in English, which is the official language in Botswana.A demographic information questionnaire was used to gather such information as age, sex, and school level.Anthropometric measures including height, weight, waist and hip circumferences were taken.Daily PA and sedentary behavior were assessed using self-report items from the Youth Risk Behavior Surveillance System (YRBSS) . ParticiSedentary behavior was also assessed using self-report items from the YRBSS . Partici2), and participants were assigned to the appropriate BMI category for age using the World Health Organization (WHO) classification: underweight (BMI < 18.50 kg/m2), normal weight (BMI 18.50–24.99 kg/m2), overweight (BMI ≥ 25 kg/m2), and obese (BMI ≥ 30 kg/m2) [Body weight was measured to the nearest 0.1 kg in light clothing using a portable Tanita digital scale . Height was measured without shoes using a portable stadiometer  accurate to 1 mm. BMI was calculated (weight in kg/height in m0 kg/m2) .Body weight satisfaction (BWS) was determined as a congruence score between participants’ self-rating on body size (perceived body size) and their BMI classification based on WHO cut-off points . This apPASE was assessed using seven items adapted from a PA Self-Efficacy Scale . The selPermission to conduct this research was obtained from the Ethics Committees of the Ministry of Health in Botswana. IRB approvals were obtained from the collaborating institutions in Botswana. In addition to providing assent, all participants under 21 years had to provide signed consent from parents or legal guardians to participate in the study. Those aged 21 years and above, which is considered legal age of consent in Botswana, offered their own verbal and written consent. All the participants were required to provide verbal and signed assent/consent prior to completing questionnaires. Individuals and their parents/guardians were made aware of their rights to decline participation, withdraw from the study at any point, or decline to answer any questions.Descriptive statistics for demographic and various health indicators are presented, and independent samples t-tests and chi-square analyses were conducted to test for differences as a function of HIV status and sex. A multivariate analysis of variance (MANOVA) tested for simultaneous differences in a set of three dependent variables assessing physical activity behavior  by HIV status and sex. Given the potential age and BMI differences, these were included in the model as covariates. An examination of the Box’s M test showed that homogeneity of variance-covariance matrices was not violated. A second MANOVA was used to examine if PASE, BWS, and enjoyment of PA differ as a function of HIV status, controlling for age, sex, and BMI. A subsequent analysis was also conducted by repeating this MANOVA with the absolute value of BWS used as a dependent variable to identify if the direction of the discrepancy impacted the results. Multiple regression models were then run to test for the association of PASE, BWS, and enjoyment of PA with each of daily PA, muscle strengthening activities, and sedentary behavior. Age, sex, and BMI were entered simultaneously in the models to control for their effects on daily PA, muscle strengthening activities, and sedentary behavior. To avoid the inflated likelihood of a Type I error due to multiple testing, the significance level of each test  was adjusted using the Bonferroni correction that divides the significance level by the number of tests, resulting in .017 (= .05/3). An inspection of model assumptions indicated that linearity, normality, and multicollinearity assumptions were met for all model analyses. The data were analyzed using the Statistical Package for Social Sciences (SPSS) (Version 24).Demographic and health variables were examined to consider differences as a function of HIV status and sex. The data showed that the groups were similar across many of these variables. However, the HIV- group was significantly younger than the HIV+ group. The HIV- group was also taller, and heavier than the HIV+ group, but there were no significant differences in BMI. The HIV+ had more cases of diabetes and asthma than the HIV- group. In this sample, the females were significantly younger than the males. They were also significantly shorter and lighter than the males but did not have significantly different BMI. A summary of this information is presented in F = 1.36, p>.05. However, there were significant main effects for HIV status, F = 3.18, p < .05, F = 12.27, p < .05, p < .05, and for muscle strengthening activities, p < .01. The mean differences showed that the HIV- group  had significantly higher daily PA than the HIV+ group . The HIV- group  also had a significantly higher mean for muscle strengthening activities than the HIV+ group . Univariate analyses for sex indicated that the only significant effect was for muscle strengthening activities with males  reporting more muscle strengthening activities than females . A summary of the mean differences by HIV status and sex is presented in To address the first objective of the study, a two-way MANOVA was run to determine differences in daily PA, muscle strengthening activities, and sedentary behavior by HIV status and sex, controlling for age and BMI. The results indicated that there was not a significant interaction effect, F = 6.49, p< .001, p < .001, and for BWS, p < .05. An examination of means for PASE by HIV status showed a higher mean for HIV- participants  compared to HIV+ participants . Results for BWS indicated that HIV- participants  reported a more negative BWS than did HIV+ participants .The MANOVA indicated that there was a statistically significant multivariate main effect for HIV status, F = 5.80, p < .01, p < .001.A subsequent MANOVA was conducted using the absolute value of BWS instead of BWS to see if the previous finding was driven by the direction of the discrepancy rather than the magnitude of the discrepancy. The results yielded a statistically significant multivariate main effect for HIV status, Multiple regression analyses were run to address the third of objective of this study regarding whether PASE, BWS, and enjoyment of PA are associated with daily PA, muscle strengthening activities, and sedentary behavior, controlling for age, sex, and BMI. Age, sex, and BMI were simultaneously entered in model 1 of the regression followed by the three psychosocial correlates in model 2. The results are summarized in F = 6.55, p < .001, adjusted R2= .13. Higher PASE  and greater enjoyment of PA  were predictive of higher daily PA. Both model 1 , F = 11.15, p < .001, adjusted R2 = .11, and model 2 , F = 12.60, p < .001, adjusted R2 = .23, were statistically significant predictors of muscle strengthening activities. Males reported more muscle strengthening activities than females , while an increase in age was associated with a decline in muscle strengthening activities . Higher PASE  and enjoyment of PA  were predictive of greater participation in muscle strengthening activities. Only the covariates were significant predictors of variance in sedentary behaviors, F = 4.12, p < .01, adjusted R2 = .05. Age significantly predicted sedentary behavior , indicating that older age was associated with more sedentary behavior.After controlling for age, sex, and BMI in model 1, PASE, BWS, and enjoyment of PA explained a significant portion of the variance in daily PA, There is limited research evidence related to psychosocial correlates and determinants of PA and sedentary behavior among youth who are not middle or upper-class and from western countries. Furthermore, there is no evidence on these relationships among adolescents living with HIV. Given the known benefits of PA participation to overall health and wellbeing, such information would be valuable to PA and health promotion among this group. Therefore, we sought to examine if daily PA, muscle strengthening activities, and sedentary behavior differ by HIV status and sex while controlling for age and BMI. Second, we wanted to determine if psychosocial correlates of PA  differ as a function of HIV status while controlling for age, sex and BMI. Finally, we wanted to determine if psychosocial factors are associated with participation in daily PA, muscle strengthening activities, and sedentary behavior among youth in our study while controlling for their age, sex, and BMI.Descriptive data shows that except for being shorter, lighter, older, and reporting more cases of diabetes and asthma, the HIV+ group did not differ significantly from the HIV- group on selected demographic variables and indicators of overall health and wellbeing. Importantly, given the differences in height, weight, and age, all analyses were conducted controlling for BMI and age.Results from this study show that males spent significantly more time in muscle strengthening activities than females. They also reported a marginally higher involvement in daily PA compared to females. These significant sex differences are noteworthy because they are consistent with previous findings on PA and muscle strengthening activities ,13,32. PNo significant differences were found on sedentary behavior by sex or HIV status, however, sedentary behavior significantly increased with age. This finding is consistent with what has been reported about the growing prevalence of sedentary behavior across children and youth populations ,43. It aResults from this study showed that the HIV- group had higher daily PA, muscle strengthening activities and PASE, but lower BWS than the HIV+ group. The results also showed a positive relationship between PASE, enjoyment of PA and higher participation in daily PA and muscle strengthening activities. This suggests improvements in PASE might benefit PA behavior, especially among HIV+ youth. The positive relationship between PASE and higher participation in daily PA and muscle strengthening activities confirms findings from previous studies, which suggests that attitudes and efficacy beliefs are indeed important predictors as well as mediators of PA behavior ,26,42,45Although, this study’s findings on the direction and magnitude of BWS were not consistent, they may have implications relative to body image and body dissatisfaction issues reported in previous studies ,44,47,48The current findings suggest that more research is needed to understand the role of psychosocial factors like PASE, BWS, and enjoyment of PA on PA and health behaviors of youth living with HIV. The differences on daily PA and muscle strengthening activities between HIV+ and HIV- participants suggest more research is needed to understand the extent to which the disease and being on ART limits or alters the trajectories of PA and muscle strengthening activities for individuals living with HIV. Another important consideration is the impact of culture, gender roles, and body stereotypes on PA behaviors for individuals living with HIV and on ART. Evidence of emotional challenges associated with peripheral fat loss and central fat gain due to ART, reported elsewhere, suggest this is an issue worthy of investigation . An explThere are several limitations worth noting in the current study. First the cross-sectional nature of our study means we cannot determine a causal relationship between study variables. Although we have discussed these findings as suggesting that PASE and enjoyment of PA predict PA and muscle strengthening activities, the opposite is equally possible. Although the HIV+ group was assessed in a previous study in which they were followed for a few years, data from this study is truly cross-sectional. A time series longitudinal design allowing periodic assessments of this group and their counterparts who are HIV- would certainly allow for stronger conclusions. This line of research could also benefit from experimental and mixed-method designs.Another limitation of our study is that our data is based solely on self-report measures. Given the known limitations of self-reported PA, the use of objective measures of PA and muscle strengthening activities is likely to enhance the quality of PA assessment. Although the YRBSS and the PASE items have been used quite extensively and produced highly consistent results with different sub-populations, there may be cultural and other contextual limitations to these tools that may affect the quality of responses. Furthermore, we cannot rule out bias from recall and social desirability. Finally, investigations that examine other possible barriers and facilitators of PA, sedentary behavior, and enjoyment of PA in this context and among HIV+ youth are likely to provide additional insights. Objective measures may be needed to verify self-report specially to determine muscular strengthening, enjoyment of PA and BWS. The role of social influencers such as parents, peers, school, and the community should offer a more comprehensive picture of the PA environment. The weak result on enjoyment of PA in the study, when it is generally associated with increased PA and muscle strengthening activities, suggests a need for a more robust measure of this variable.Overall the current study demonstrates the relationship between PASE, daily PA and muscle strengthening activities. Further, the study shows the potential role that PASE, enjoyment of PA, and BWS could play in the promotion of PA among HIV+ youth. The findings have implications for interventions and clinical research into how PA and exercise could be used to mitigate the effects of living with chronic conditions such as HIV. The findings suggest the possibility of some socio-cultural nuances related to definitions of body weight, body size, and enjoyment of PA that are worthy of further investigation. Studying such nuances may be necessary to understand and develop appropriate strategies to promote healthy PA behaviors and reduce barriers to PA and exercise. One of the strengths of this study is that it offers much needed evidence on correlates of PA and sedentary behavior among a less studied subpopulation of youth. These are HIV+ youth on ART and from an African country. It also makes a comparison of these youth with those who are HIV-. The comparative evidence provided in this study is particularly relevant to the development of a global understanding of PA as it relates to health and wellbeing of youth across the board, especially those living with HIV and other chronic conditions.S1 Dataset(SAV)Click here for additional data file.