Exposure to environmental pollutants and genetic variants related to oxidative stress and xenobiotic metabolism-Association with prostate cancer.

This study assessed whether genetic variants coding for certain enzymes involved in xenobiotic detoxification, antioxidant defences and DNA repair, along with exposure to environmental chemicals, were associated with an increased prostate cancer (PCa) risk. The study population consisted of 300 men (150 PCa cases and 150 controls) which underwent prostate biopsy as their serum prostate specific antigen (PSA) levels were greater than 4 ng/ml. Genetic variants in GSTM1, GSTP1, SOD2, CAT, GPX1, XRCC1 were determined and data for chemical exposures was obtained through a structured questionnaire and by biomonitoring in a subsample of cases and controls. High serum PSA levels were associated with a greater risk of PCa, while physical exercise appears to exert a protective effect against its development. In addition, elevated urinary levels of certain organic pollutants, such as benzo(a)pyrene (BaP), bisphenol A (BPA), and ethyl-paraben (EPB), were associated with an increased risk of PCa.

New analytical method for the determination of endocrine disruptors in human faeces using gas chromatography coupled to tandem mass spectrometry.

Endocrine-disrupting chemicals are environmental pollutants that can enter our bodies and cause diverse pathologies. Some bisphenols and parabens have been shown to be capable of modifying proper functioning of the endocrine system. Among other dysfunctions, endocrine-disrupting chemicals can cause changes in intestinal microbiota. Faeces are a convenient matrix that can be useful for identifying the quantity of endocrine disruptors that reach the intestine and the extent to which the organism is exposed to these pollutants. The present work developed a new analytical method to determine 17 compounds belonging to the paraben and bisphenol families found in human faeces. The extraction method was optimized using an ultrasound-assisted extraction technique followed by a clean-up step based on the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) technique. Optimization was performed using the design of experiments technique. In validation analysis, the method was proven to be linear over a wide range. R-squared outcomes were between 95 and 99%. Selectiveness and sensitivity outcomes were acceptable, with detection limits being between 1 and 10 ng g-1 in all cases, whilst quantification limits were between 3 and 25 ng g-1 in all instances, with the exception of bisphenol AF. The method was deemed accurate, with recovery values being close to 100% and relative standard deviations being lower than 15% in all cases. Applicability was examined by analysing 13 samples collected from volunteers (male and female). All samples were contaminated with at least one of the analytes studied. The most commonly found compounds were methylparaben and bisphenol A, which were detected in almost all samples and quantitatively determined in 11 and 12 samples, respectively. Of the 17 compounds analysed, 11 were found in at least one sample. Outcomes demonstrate that faeces can be a good matrix for the determination of exposure to contaminants of interest here.

Levels of Bisphenol A and its analogs in nails, saliva, and urine of children: a case control study.

Introduction: A growing number of studies link the increase in overweight/obesity worldwide to exposure to certain environmental chemical pollutants that display obesogenic activity (obesogens). Since exposure to obesogens during the first stages of life has been shown to have a more intense and pronounced effect at lower doses, it is imperative to study their possible effects in childhood. The objective here was to study the association of Bisphenol A (BPA) and 11 BPA analogs in children, using three biological matrices (nails, saliva and urine), and overweight and obesity (n = 160). Methods: In this case-control study, 59 overweight/obese children and 101 controls were included. The measuring of Bisphenols in the matrices was carried out by ultra-high performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UHPLC-MS/MS). Logistic regression was used to study the association between overweight/obesity and Bisphenol exposure. Results: The results suggested that BPF in nails is associated with overweight/ obesity in children (OR:4.87; p = 0.020). In saliva, however, the highest detected concentrations of BPAF presented an inverse association (OR: 0.06; p = 0.010) with overweight/obesity. No associations of statistical significance were detected between exposure to BPA or its other analogs and overweight/obesity in any of the biological matrices.

Bioconcentration of pharmaceuticals in benthic marine organisms (Holothuria tubulosa, Anemonia sulcata and Actinia equina) exposed to environmental contamination by atenolol and carbamazepine.

The present work assess the bioconcentration kinetics of atenolol (ATN) and carbamazepine (CBZ) in common marine organisms including Holothuria tubulosa, Anemonia sulcata and Actinia equina under controlled laboratory conditions. CBZ exhibited higher uptake and excretion rates resulting higher bioconcentration factor (BCF) (41-537 L/kg for CBZ vs 7-50 L/kg for ATN) although both are below the limits established by the European Union (EU). The measured BCF using kinetic data showed some differences with those predicted using the concentrations measured at the steady-state, probably explained because the steady state was not ready reached. The animal-specific BCF followed the order of Holothuria tubulosa > Actinia equina > Anemonia sulcata for ATN while was the opposite for CBZ. The study highlighted between-tissues differences in the digestive tract and the body wall of the Holothuria tubulosa. The work presented is the first to model bioconcentration of ATN and CBZ in holothurian and anemone animal models.

Analytical method for the determination of usually prescribed antibiotics in human nails using UHPLC-MS/MS. Comparison of the efficiency of two extraction techniques.

Antibiotics are a group of drugs used for the treatment of bacterial diseases. They are used in both human and veterinary medicine and, although they are not permitted, they are sometimes used as growth promoters. The present research compares two extraction techniques: ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) in order to evaluate their efficiency in the determination of 17 usually prescribed antibiotics in human nails. The extraction parameters were optimized using multivariate techniques. Once both techniques were compared, MAE was selected as optimal due to its greater experimental practicability together with the better extraction efficiencies it provides. Target analytes were detected and quantified by ultra-high performance liquid chromatography with tandem mass spectrometry detection (UHPLC-MS/MS). The run time was 20 min. The methodology was then successfully validated, obtaining acceptable analytical parameters according to the guide used. Limits of detection were between 0.3 and 3 ng g-1 and limits of quantification were in the range from 1.0 to 4.0 ng g-1. Recovery percentages ranged from 87.5% to 114.2%, and precision (in terms of standard deviation) was less than 15% in all cases. Finally, the optimized method was applied to nails taken from 10 volunteers and the results revealed the presence of one or more antibiotics in all the samples examined. The most commonly found antibiotic was sulfamethoxazole, followed by danofloxacin and levofloxacin. The results demonstrated, on the one hand, the presence of these compounds in the human body and, on the other hand, the suitability of nails as a non-invasive biomarker of exposure.

Accumulation and metabolization of the antidepressant venlafaxine and its main metabolite o-desmethylvenlafaxine in non-target marine organisms Holothuria tubulosa, Anemonia sulcata and Actinia equina.

The assessment of exposure to the antidepressant venlafaxine and its major metabolite o-desmethylvenlafaxine in Holothuria tubulosa, Anemonia sulcata and Actinia equina is proposed. A 28-day exposure experiment (10 µg/L day) followed by a 52-day depuration period was conducted. The accumulation shows a first-order kinetic process reaching an average concentration of 49,125/54342 ng/g dw in H. tubulosa and 64,810/93007 ng/g dw in A. sulcata. Venlafaxine is considered cumulative (BCF > 2000 L/kg dw) in H. tubulosa, A. sulcata and A. equina respectively; and o-desmethylvenlafaxine in A. sulcata. Organism-specific BCF generally followed the order A. sulcata > A. equina > H. tubulosa. The study revealed differences between tissues in metabolizing abilities in H. tubulosa this effect increases significantly with time in the digestive tract while it was negligible in the body wall. The results provide a description of venlafaxine and o-desmethylvenlafaxine accumulation in common and non-target organisms in the marine environment.

Uptake and depuration of three common antibiotics in benthic organisms: Sea cucumber (Holothuria tubulosa), snakelocks anemone (Anemonia sulcata) and beadlet anemone (Actinia equina).

Antibiotics are widely used drugs in human and veterinary medicine, which has attracted great attention in relation to the development of bacterial resistance, currently a problem of great concern for governments and states, as it is related to the resurgence of infectious diseases already eradicated. Understanding the bioaccumulation of antibiotics in aquatic organisms is an important key to understanding their risk assessment. The present study was designed to study the bioaccumulation of target antibiotics in relevant organisms inhabiting benthic marine environments. The uptake and elimination of ciprofloxacin (CIP), sulfamethoxazole (SMX) and trimethoprim (TMP) were investigated in sea cucumbers (Holothuria tubulosa), snakelock anemone (Anemonia sulcata) and beadlet anemone (Actinia equina) under controlled laboratory conditions. The results show that antibiotics have a particular trend over time during all periods of absorption and depuration. The tissue distribution of antibiotics in sea cucumber is strongly influenced by the structure of the compounds, while CIP is concentrated in the body wall; TMP is concentrated in the digestive tract. Two different approaches were used to estimate bioconcentration factors (BCFs) in different animal models, based on toxicokinetic data and measured steady-state concentrations. The BCF ranges were 456-2731 L/kg, 6-511 L/kg and 9-100 L/kg for TMP, CIP and SMX, respectively. The estimated BCF values obtained classify TMP as cumulative in A. equina and H. tubulosa, underlining the potential bioconcentration in these marine organisms. A correlation was observed between the BCFs of the target antibiotics and the octanol-water distribution coefficient (Dow) (r2 > 0.7). The animal-specific BCF followed the order of beadlet anemone > sea cucumber > snakelock anemone.

Presence of Parabens in Different Children Biological Matrices and Its Relationship with Body Mass Index.

Parabens have been accepted almost worldwide as preservatives by the cosmetic, food, and pharmaceutical industries. Since epidemiological evidence of the obesogenic activity of parabens is weak, the aim of this study was to investigate the association between parabens exposure and childhood obesity. Four parabens (methylparaben/MetPB, ethylparaben/EthPB, propylparaben/PropPB, and butylparaben/ButPB) were measured in 160 children's bodies between 6 and 12 years of age. Parabens measurements were performed with ultrahigh-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). Logistic regression was used to evaluate risk factors for elevated body weight associated with paraben exposure. No significant relation was detected between children's body weight and the presence of parabens in the samples. This study confirmed the omnipresence of parabens in children's bodies. Our results could be a basis for future research about the effect of parabens on childhood body weight using nails as a biomarker due to the ease of its collection and its non-invasive character.

Bioaccumulation/bioconcentration of pharmaceutical active compounds in aquatic organisms: Assessment and factors database.

There is increasing evidence that the presence of certain pharmaceuticals in the environment leads to biota exposure and constitute a potential risk for ecosystems. Bioaccumulation is an essential focus of risk assessment to evaluate at what degree emerging contaminants are a hazard both to the environment and the individuals that inhabit it. The main goals of the present review are 1) to summarize and describe the research and factors that should be taken into account in the evaluation of bioaccumulation of pharmaceuticals in aquatic organisms; and 2) to provide a database and a critical review of the bioaccumulation/bioconcentration factors (BAF or BCF) of these compounds in organisms of different trophic levels. Most studies fall into one of two categories: laboratory-scale absorption and purification tests or field studies and, to a lesser extent, large-scale, semi-natural system tests. Although in the last 5 years there has been considerable progress in this field, especially in species of fish and molluscs, research is still limited on other aquatic species like crustaceans or algae. This revision includes >230 bioconcentration factors (BCF) and >530 bioaccumulation factors (BAF), determined for 113 pharmaceuticals. The most commonly studied is the antidepressant group, followed by diclofenac and carbamazepine. There is currently no reported accumulation data on certain compounds, such as anti-cancer drugs. BCFs are highly influenced by experimental factors (notably the exposure level, time or temperature). Field BAFs are superior to laboratory BCFs, highlighting the importance of field studies for reliable assessments and in true environmental conditions. BAF data appears to be organ, species and compound-specific. The potential impact on food web transfer is also considered. Among different aquatic species, lower trophic levels and benthic organisms exhibit relatively higher uptake of these compounds.

Presence of parabens in children's faeces. Optimization and validation of a new analytical method based on the use of ultrasound-assisted extraction and liquid chromatography-tandem mass spectrometry.

Due to their antimicrobial properties, parabens are a family of synthetic chemical compounds widely used as preservative additives in food and cosmetics. For this reason, humans are highly exposed to them. These substances are capable of altering the proper functioning of the endocrine system and are classified as endocrine disrupting chemicals (EDCs). Traditionally, urine has been the typical matrix studied as an excretion route. However, faeces contain valuable information. In the present study, the presence of methyl-, ethyl-, isopropyl-, propyl-, isobutyl-, butyl- and phenylparaben in stool samples from children has been evaluated. A new analytical method has been optimised and validated. The method is based on the use of ultrasound-assisted extraction followed by clean-up of the extracts by dispersive solid phase extraction dSPE). Parabens were analysed by ultrahigh performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). The matrix effect was evaluated and a significant effect was observed for all analytes. Therefore, calibration and validation were performed by addition of different concentrations of analytes to faecal blanks. The coefficient of determination (%R2) for calibration curves was higher than 98.9% in all cases. The limits of detection and quantification were between 0.2 and 0.4 and 0.6-1.0 ng g-1 respectively. The recovery for accuracy assessment had values between 89.0% and 112.7% with an RSD of less than 15% in all cases. The method was successfully applied to 14 samples from children volunteers, 100% of which showed contamination by at least one of the analysed compounds.

An Overview of Analytical Methods to Determine Pharmaceutical Active Compounds in Aquatic Organisms.

There is increasing scientific evidence that some pharmaceuticals are present in the marine ecosystems at concentrations that may cause adverse effects on the organisms that inhabit them. At present, there is still very little scientific literature on the (bio)accumulation of these compounds in different species, let alone on the relationship between the presence of these compounds and the adverse effects they produce. However, attempts have been made to optimize and validate analytical methods for the determination of residues of pharmaceuticals in marine biota by studying the stages of sample treatment, sample clean-up and subsequent analysis. The proposed bibliographic review includes a summary of the most commonly techniques, and its analytical features, proposed to determine pharmaceutical compounds in aquatic organisms at different levels of the trophic chain in the last 10 years.

Improved method for the determination of endocrine-disrupting chemicals in urine of school-age children using microliquid-liquid extraction and UHPLC-MS/MS.

The presence of endocrine-disrupting chemicals in our daily life is increasing every day and, by extension, human exposure and the consequences thereof. Among these substances are bisphenols and parabens. Urine is used to analyze the exposure. The determination of 12 bisphenol homologues and 6 parabens is proposed. A procedure based on a method previously developed by our research group in 2014 is improved. The extraction yield is higher, because the new protocol is 5 times more efficient. Also, a comparison between calibration with pure standards and matrix calibration, to calculate the matrix effect, was also made. A high grade of matrix effect for all analytes was observed. In terms of validation, the limits of detection (LOD) were between 0.03 and 0.3 ng mL-1 and limits of quantification (LOQ) 0.1 to 1.0 ng mL-1, respectively, and the recovery is higher than 86.4% and lower than 113.6%, with a RSD lower than 13.5% in all cases. A methodology for accurate and sensitive quantification of bisphenol homologues together with parabens in human urine using UHPLC-MS/MS was developed. The method was successfully applied to 30 urine samples from children.

Analytical methods for the determination of endocrine disrupting chemicals in cosmetics and personal care products: A review.

Personal care products (PCPs) and cosmetics are indispensable product in our daily routine. Their widespread use makes them a potential route of exposure for certain contaminants to which human would not be normally exposed. One of these contaminants includes endocrine disrupting chemicals, molecules capable of mimicking the body's natural hormones and interfering with the endocrine system. Some of them are ingredients included in the product's formulation, such as UV-filters (sunscreens), phthalates (plasticizers and preservatives), synthetic musks (fragrances), parabens and other antimicrobial agents (antimicrobial preservatives). Others are non-intended added substances that may result from the manufacturing process or migration from the plastic packaging, as with bisphenols and perfluorinated compounds. Some of these endocrine disruptors have been restricted or even banned in cosmetics and PCPs given the high risk they pose to health. Thus, the development of fast, sensitive and precise methods for the identification and quantification of these compounds in cosmetics is a substantial need in order to ensure consumer safety and provide insight into the real risk of human exposure. The present work aims at reviewing the more recently developed analytical methods published in the literature for the determination of endocrine disrupting chemicals in cosmetics and PCPs using chromatographic techniques, with a focus on sample treatment and the quality of analytical parameters.

Dietary exposure to parabens and body mass index in an adolescent Spanish population.

Parabens are alkyl esters of p-hydroxybenzoic acid which are extensively used in cosmetics, pharmaceuticals and foodstuffs due to their antimicrobial properties. The most commonly used parabens are methyl-(MeP), ethyl-(EtP), propyl-(PrP) and butyl-(BuP) paraben. Most human exposure to parabens is achieved through the consumption of food or pharmaceutical products and the use of personal care products. However, studies on dietary parabens exposure and the associated factors are very scarce. The main aim of the present study was to explore factors associated with dietary exposure to parabens in Spanish adolescents according to gender. Dietary data and anthropometric measures were collected from 585 adolescents (53.4% boys) aged 12-16 years. Parabens exposure through diet was assessed using a food frequency questionnaire with food products providing more than 95% of energy and macronutrient intake being included in analysis. Stepwise regression was used to identify the foods that most contributed to parabens intake. Logistic regression was used to evaluate factors predicting higher dietary exposure to parabens. The main contributors to dietary MeP, EtP, PrP and BuP exposure in adolescent boys were eggs (41.9%), canned tuna (46.4%), bakery and baked goods products (57.3%) and pineapple (61.1%). In adolescent girls, the main contributors were apples and pears (35.3%), canned tuna (42.1%), bakery and baked goods products (55.1%) and olives (62.1%). Overweight/obese girls were more likely to belong to the highest tertile of overall parabens intake (odds ratio [OR]: 3.32; 95% confidence interval [95% CI]: 1.21-9.15) and MeP (OR: 3.05; 95% CI: 1.14-8.12) than those with a body mass index lower than 25 kg/m2. These findings suggest a positive association between dietary exposure to parabens and overweight/obesity in adolescent girls.

Factors Associated with Exposure to Dietary Bisphenols in Adolescents.

Obesogenic endocrine-disrupting chemicals, such as bisphenol A (BPA) and its analogue bisphenol S (BPS), seem to play an important role in the development of obesity, although contradictory results have been reported. The aim of the present study was to conduct a gender analysis of the factors associated with exposure to dietary bisphenols in 585 Spanish adolescents. Dietary BPA and BPS exposure was assessed using a food frequency questionnaire. Foods and macronutrients accounting for more than 95% of energy intake were selected for analysis. Stepwise regression was used to estimate the foods that most contributed to dietary bisphenol exposure in the sample. Gender-related factors associated with greater dietary bisphenol exposure were evaluated using multivariate logistic regression models. Canned tuna was the main dietary source of BPA and BPS in both adolescent boys and girls. Overweight/obese girls showed a higher risk of high dietary exposure to BPA (odds ratio (OR): 3.38, 95% confidence interval (CI): 1.25-9.07) and total bisphenols (OR: 2.81, 95% CI: 1.03-7.67) in comparison with girls with a BMI lower than 25 kg/m2. Present results indicate a positive association of dietary exposure to both total bisphenols and BPA with being overweight/obese in adolescent girls.

Presence of Parabens and Bisphenols in Food Commonly Consumed in Spain.

Given the widespread use of bisphenols and parabens in consumer products, the assessment of their intake is crucial and represents the first step towards the assessment of the potential risks that these compounds may pose to human health. In the present study, a total of 98 samples of food items commonly consumed by the Spanish population were collected from different national supermarkets and grocery stores for the determination of parabens and bisphenols. Our analysis demonstrated that 56 of the 98 food samples contained detectable levels of parabens with limits of quantification (LOQ) between 0.4 and 0.9 ng g-1. The total concentration of parabens (sum of four parabens: ∑parabens) ranged from below the LOQ to 281.7 ng g-1, with a mean value of 73.86 ng g-1. A total of 52% of the samples showed detectable concentrations of bisphenols. Bisphenol A (BPA) was the most frequently detected bisphenol in the food samples analysed, followed by bisphenol S (BPS) and bisphenol E (BPE). Bisphenol AF (BPAF), bisphenol B (BPB) and bisphenol P (BPP) were not found in any of the analysed samples. LOQ for these bisphenols were between 0.4 and 4.0 ng g-1.

Determination of ultraviolet filters in human nails using an acid sample digestion followed by ultra-high performance liquid chromatography-mass spectrometry analysis.

Ultraviolet filters (UV-filters) are specific chemicals that absorb and reflect UVA and UVB radiation from the sun. They are regularly used in sunscreens and in other personal care products (PCPs), and in products like plastics, adhesives, toys, or furniture finishes. This work develops and validates a new method to determine concentrations of UV-filters (BP-1, BP-2, BP-3, BP-6, BP-8, 4-OH-BP, THB, AVB) in human nail samples. Nails are easily available and are considered to be suitable indicators of cumulative and continued exposure to harmful chemicals. The treatment of nail samples includes microwave assisted digestion/extraction (MAE) in a methanolic solution of o-phosphoric acid (0.05 mol L-1) followed by analyte determination using ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS/MS) in multiple reaction monitoring mode. The analytes were separated in less than 10 min. The digestion procedure was optimized using multivariate techniques. Matrix-matched calibration with a pig hoof matrix was used for validating the method. A study of accuracy with spiked blank samples was also conducted. The calculated detection limits varied between 0.2 and 1.5 ng g-1, and quantification limits between 1.0 and 5.0 ng g-1. The trueness of the method was an estimation of the recovery, which was between 90.2% and 112.2%; with an estimated precision (relative standard deviation, % RSD) lower than 12.3% for all UV-filters. Nail samples were obtained from 22 volunteers (male and female). The results showed that BP-1 and BP-3 mainly bioaccumulate in human nails.

Assessing bioaccumulation potential of personal care, household and industrial products in a marine echinoderm (Holothuria tubulosa).

A bioaccumulation study of 16 emerging contaminants including preservatives, UV-filters, biocides, alkylphenols, anionic surfactants and plasticizers, in Holothuria tubulosa Gmelin, 1791 specimens was developed. Water and sediments from their coastal habitat were also analyzed. Sediment-water distribution coefficients (log Kd) were in the range 0.78 to 2.95. A rapid uptake and bioaccumulation of pollutants was found. Compounds were detected in intestine and gonads of H. tubulosa after only eight days of exposure. Field-based bioconcentration (BCF) and biota-sediment accumulation factors (BSAF) were calculated. Log BCF > 1 were obtained for most of the compounds studied, indicating their tendency to accumulate in tissue of H. Tubulosa. BCF values decrease as follow: Triclocarban > anionic surfactants > benzophenone 3 > non-ionic surfactants > bisphenol A > parabens. These data provide a detailed accounting of the distribution patterns of some emerging contaminants in organisms at the lower trophic level, representing a potential source of contaminants for organisms in higher levels of the food chain.

Determination of endocrine disrupting chemicals in human nails using an alkaline digestion prior to ultra-high performance liquid chromatography-tandem mass spectrometry.

Rapid industrialization has resulted in a progressive increase in human exposure to hazardous chemicals. The present work develops and validates a new method to determinate 18 endocrine disrupting chemicals (EDCs) in human nail samples. In contrast to other common biological samples, nail sampling is non-invasive and since they take several months to grow out, they are well suited for measuring and reflecting the cumulative exposure to harmful substances in the long term. A digestion of samples with a 0.04 M solution of sodium hydroxide is carried out followed by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), working in multiple-reaction-monitoring (MRM) mode. The compounds were separated in 8 min. Multivariate optimization strategies were used for the optimization of the parameters that affects the digestion procedure. The validation was developed using a matrix-matched calibration and a recovery assay with spiked samples. The limits of detection and quantification ranged from 0.3 to 1.2 ng g-1 and from 1 to 5 ng g-1, respectively. Recovery rates for spiked samples were between 88% and 113% and the relative standard deviation (% RSD) was lower than 12.7% for all studied EDCs. The method was applied for the analysis of these compounds in human nail samples from volunteers. All samples tested positive for several of the analyzed EDCs.

Bisphenol A Analogues in Food and Their Hormonal and Obesogenic Effects: A Review.

Bisphenol A (BPA) is the most well-known compound from the bisphenol family. As BPA has recently come under pressure, it is being replaced by compounds very similar in structure, but data on the occurrence of these BPA analogues in food and human matrices are limited. The main objective of this work was to investigate human exposure to BPA and analogues and the associated health effects. We performed a literature review of the available research made in humans, in in vivo and in vitro tests. The findings support the idea that exposure to BPA analogues may have an impact on human health, especially in terms of obesity and other adverse health effects in children.