Early adversity and insulin: neuroendocrine programming beyond glucocorticoids.

Exposure to direct or contextual adversities during early life programs the functioning of the brain and other biological systems, contributing to the development of physical as well as mental health issues in the long term. While the role of glucocorticoids in mediating the outcomes of early adversity has been explored for many years, less attention has been given to insulin. Beyond its metabolic effects in the periphery, central insulin action affects synaptic plasticity, brain neurotransmission, and executive functions. Knowledge about the interactions between the peripheral metabolism and brain function from a developmental perspective can contribute to prevention and diagnosis programs, as well as early interventions for vulnerable populations.

Dopamine D4 receptor gene polymorphism (DRD4 VNTR) moderates real-world behavioural response to the food retail environment in children.

BACKGROUND: Evidence for the impact of the food retailing environment on food-related and obesity outcomes remains equivocal, but only a few studies have attempted to identify sub-populations for whom this relationship might be stronger than others. Genetic polymorphisms related to dopamine signalling have been associated with differences in responses to rewards such as food and may be candidate markers to identify such sub-populations. This study sought to investigate whether genetic variation of the dopamine D4 receptor gene (DRD4 exon III 48 bp VNTR polymorphism) moderated the association between local exposure to food retailers on BMI and diet in a sample of 4 to12-year-old children. METHODS: Data collected from a birth cohort and a community cross-sectional study conducted in Montreal, Canada, were combined to provide DRD4 VNTR polymorphism data in terms of presence of the 7-repeat allele (DRD4-7R) for 322 children aged between 4 and 12 (M (SD): 6.8(2.8) y). Outcomes were Body Mass Index (BMI) for age and energy density derived from a Food Frequency Questionnaire. Food environment was expressed as the proportion of local food retailers classified as healthful within 3 km of participants' residence. Linear regression models adjusted for age, sex, income, cohort, and geographic clustering were used to test gene*environment interactions. RESULTS: A significant gene*food environment interaction was found for energy density with results indicating that DRD4-7R carriers had more energy dense diets than non-carriers, with this effect being more pronounced in children living in areas with proportionally more unhealthy food retailers. No evidence of main or interactive effects of DRD4 VNTR and food environment was found for BMI. CONCLUSIONS: Results of the present study suggest that a genetic marker related to dopamine pathways can identify children with potentially greater responsiveness to unhealthy local food environment. Future studies should investigate additional elements of the food environment and test whether results hold across different populations.

Predicted DRD4 prefrontal gene expression moderates snack intake and stress perception in response to the environment in adolescents.

Body weight is substantially determined by eating behaviors, which are themselves driven by biological factors interacting with the environment. Previous studies in young children suggest that genetic influences on dopamine function may confer differential susceptibility to the environment in such a way that increases behavioral obesity risk in a lower socioeconomic status (SES) environment but decreases it in a higher SES environment. We aimed to test if this pattern of effect could also be observed in adolescence, another critical period for development in brain and behavior, using a novel measure of predicted expression of the dopamine receptor 4 (DRD4) gene in prefrontal cortex. In a sample of 76 adolescents (37 boys and 39 girls from Baltimore, Maryland/US, aged 14-18y), we estimated individual levels of DRD4 gene expression (PredDRD4) in prefrontal cortex from individual genomic data using PrediXcan, and tested interactions with a composite SES score derived from their annual household income, maternal education, food insecurity, perceived resource availability, and receipt of public assistance. Primary outcomes were snack intake during a multi-item ad libitum meal test, and food-related impulsivity assessed using a food-adapted go/no-go task. A linear regression model adjusted for sex, BMI z-score, and genetic ethnicity demonstrated a PredDRD4 by composite SES score interaction for snack intake (p = 0.009), such that adolescents who had lower PredDRD4 levels exhibited greater snack intake in the lower SES group, but lesser snack intake in the higher SES group. Exploratory analysis revealed a similar pattern for scores on the Perceived Stress Scale (p = 0.001) such that the low PredDRD4 group reported higher stress in the lower SES group, but less stress in the higher SES group, suggesting that PredDRD4 may act in part by affecting perceptions of the environment. These results are consistent with a differential susceptibility model in which genes influencing environmental responsiveness interact with environments varying in obesogenicity to confer behavioral obesity risk in a less favorable environment, but behavioral obesity protection in a favorable one.

Multi-behavioral obesogenic phenotypes among school-aged boys and girls along the birth weight continuum.

Evidence shows that extremes of birth weight (BW) carry a common increased risk for the development of adiposity and related cardiovascular diseases, but little is known about the role of obesogenic behaviors in this process. Moreover, no one has empirically examined whether the relationship between BW, obesogenic behaviors and BMI along the full low-to-high birthweight continuum reflects the U-shape pattern expected from common risk at both BW extremes. Our objective was to characterize physical activity, screen time, and eating behavior and their relationship to BMI as a function of BW among school-aged boys and girls. In this cross-sectional study, 460 children aged 6 to 12 years (50% boys) from Montreal, Canada provided information on sleeping time, screen time, physical activity levels, eating behavior (emotional, external and restrained eating) and anthropometrics (height, weight, BW) through parent reported questionnaires. BMI was normalized using WHO Standards (zBMI), and BW expressed as ratio using Canadian population standards (BW for gestational age and sex). Analyses were conducted using generalized linear models with linear and quadratic terms for BW, stratified by sex and adjusted for age, ethnicity and household income. In boys, physical activity and screen time showed U-shaped associations with BW, while physical activity had an inverted U-shaped in girls. Emotional and restrained eating had positive linear relations with BW in boys and girls. Sleep time and external eating were not associated with BW. A U-shaped relationship between BW and zBMI was found in boys but no association was found in girls. Only sleep (in boys and girls), and emotional eating (girls only) were related to zBMI and mediation of the BW-zBMI relationship was only supported for emotional eating. In conclusion, BW relates to obesogenic behaviors and BMI in both non-linear and linear ways, and these associations differed by sex.

Both food restriction and high-fat diet during gestation induce low birth weight and altered physical activity in adult rat offspring: the "Similarities in the Inequalities" model.

We have previously described a theoretical model in humans, called "Similarities in the Inequalities", in which extremely unequal social backgrounds coexist in a complex scenario promoting similar health outcomes in adulthood. Based on the potential applicability of and to further explore the "similarities in the inequalities" phenomenon, this study used a rat model to investigate the effect of different nutritional backgrounds during gestation on the willingness of offspring to engage in physical activity in adulthood. Sprague-Dawley rats were time mated and randomly allocated to one of three dietary groups: Control (Adlib), receiving standard laboratory chow ad libitum; 50% food restricted (FR), receiving 50% of the ad libitum-fed dam's habitual intake; or high-fat diet (HF), receiving a diet containing 23% fat. The diets were provided from day 10 of pregnancy until weaning. Within 24 hours of birth, pups were cross-fostered to other dams, forming the following groups: Adlib_Adlib, FR_Adlib, and HF_Adlib. Maternal chow consumption and weight gain, and offspring birth weight, growth, physical activity (one week of free exercise in running wheels), abdominal adiposity and biochemical data were evaluated. Western blot was performed to assess D2 receptors in the dorsal striatum. The "similarities in the inequalities" effect was observed on birth weight (both FR and HF groups were smaller than the Adlib group at birth) and physical activity (both FR_Adlib and HF_Adlib groups were different from the Adlib_Adlib group, with less active males and more active females). Our findings contribute to the view that health inequalities in fetal life may program the health outcomes manifested in offspring adult life (such as altered physical activity and metabolic parameters), probably through different biological mechanisms.

Neurobehavioral determinants of nutritional security in fetal growth-restricted individuals.

Fetal growth restriction results from a failure to achieve a higher growth potential and has been associated with many maternal conditions, such as chronic diseases (infections, hypertension, and some cases of diabetes and obesity), exposures (tobacco smoke, drugs), and malnutrition. This early adversity induces a series of adaptive physiological responses aimed at improving survival, but imposing increased risk for developing chronic nontransmittable diseases (obesity, type II diabetes, cardiovascular disease) in the long term. Recently, mounting evidence has shown that fetal growth impairment is related to altered feeding behavior and preferences through the life course. When living in countries undergoing nutritional transition, in which individuals experience the coexistence of underweight and overweight problems (the "double burden of malnutrition"), fetal growth-restricted children can be simultaneously growth restricted and overweight-a double burden of malnutrition at the individual level. Considering food preferences as an important aspect of nutrition security, we will summarize the putative neurobiological mechanisms at the core of the relationship between fetal growth and nutrition security over the life course and the evidence linking early life adversity to later food preferences.

Association between the seven-repeat allele of the dopamine-4 receptor gene (DRD4) and spontaneous food intake in pre-school children.

BACKGROUND: Studies in adults show associations between the hypofunctional seven-repeat allele (7R) of the dopamine-4 receptor gene (DRD4), increased eating behaviour and/or obesity, particularly in females. We examined whether 7R is associated with total caloric intake and/or food choices in pre-schoolers. METHODS: 150 four-year-old children taking part in a birth cohort study in Canada were administered a snack test meal in a laboratory setting. Mothers also filled out a food frequency questionnaire to address childrens' habitual food consumption. Total caloric and individual macronutrient intakes during the snack meal and specific types of foods as reported in the food diaries were compared across 7R allele carriers vs. non-carriers, using current BMI as a co-variate. RESULTS: We found significant sex by genotype interactions for fat and protein intake during the snack test. Post hoc testing revealed that in girls, but not boys, 7R carriers ate more fat and protein than did non-carriers. Based on the food diaries, across both sexes, 7R carriers consumed more portions of ice cream and less vegetables, eggs, nuts and whole bread, suggesting a less healthy pattern of habitual food consumption. CONCLUSION: The 7R allele of DRD4 influences macronutrient intakes and specific food choices as early as four years of age. The specific pattern of results further suggests that prior associations between the 7R allele and adult overeating/obesity may originate in food choices observable in the preschool years. Longitudinal follow-up of these children will help establish the relevance of these findings for obesity risk and prevention.

Early life stress is associated with anxiety, increased stress responsivity and preference for "comfort foods" in adult female rats.

Chronic stress increases anxiety and encourages intake of palatable foods as "comfort foods". This effect seems to be mediated by altered function of the hypothalamic-pituitary-adrenal axis. In the current study, litters of Wistar rats were subjected to limited access to nesting material (Early-Life Stress group - ELS) or standard care (Control group) from postnatal day 2 to 9. In adult life, anxiety was assessed using the novelty-suppressed feeding test (NSFT), and acute stress responsivity by measurement of plasma corticosterone and ACTH levels. Preference for palatable foods was monitored by a computerized system (BioDAQ, Research Diets(®)) in rats receiving only regular chow or given the choice of regular and palatable diet for 30 days. ELS-augmented adulthood anxiety in the NSFT (increased latency to eat in a new environment; decreased chow intake upon return to the home cage) and increased corticosterone (but not ACTH) secretion in response to stress. Despite being lighter and consuming less rat chow, ELS animals ate more palatable foods during chronic exposure compared with controls. During preference testing, controls receiving long-term access to palatable diet exhibited reduced preference for the diet relative to controls exposed to regular chow only, whereas ELS rats demonstrated no such reduction in preference after prolonged palatable diet exposure. The increased preference for palatable foods showed by ELS animals may result from a habit of using this type of food to ameliorate anxiety.

Intrauterine growth restriction and the fetal programming of the hedonic response to sweet taste in newborn infants.

Intrauterine growth restriction is associated with increased risk for adult metabolic syndrome and cardiovascular disease, which seems to be related to altered food preferences in these individuals later in life. In this study, we sought to understand whether intrauterine growth leads to fetal programming of the hedonic responses to sweet. Sixteen 1-day-old preterm infants received 24% sucrose solution or water and the taste reactivity was filmed and analyzed. Spearman correlation demonstrated a positive correlation between fetal growth and the hedonic response to the sweet solution in the first 15 seconds after the offer (r = 0.864, P = 0.001), without correlation when the solution given is water (r = 0.314, P = 0.455). In fact, the more intense the intrauterine growth restriction, the lower the frequency of the hedonic response observed. IUGR is strongly correlated with the hedonic response to a sweet solution in the first day of life in preterm infants. This is the first evidence in humans to demonstrate that the hedonic response to sweet taste is programmed very early during the fetal life by the degree of intrauterine growth. The altered hedonic response at birth and subsequent differential food preference may contribute to the increased risk of obesity and related disorders in adulthood in intrauterine growth-restricted individuals.

Produção de óxido nítrico no hipocampo: investigação do envolvimento do óxido nítrico sintase neuronal no desenvolvimento de comportamentos ansiosos em animais submetidos a estresse neonatal / Hippocampal nitric oxide production: investigation of the role of neuronal nitric oxide synthase in the development of anxiety-like behaviors in animals subjected to neonatal stress

Introdução: Estudos indicam que o trauma precoce e o sistema serotoninérgico estão relacionados ao desenvolvimento de ansiedade. Esta relação poderia ser mediada pela enzima óxido nítrico sintase neuronal (nNOS), que tem papel importante no funcionamento dos receptores de serotonina. Objetivo: Investigar, através da mensuração do óxido nítrico (NO) no hipocampo, o possível envolvimento da nNOS no desenvolvimento de ansiedade em um modelo animal de adversidade no início da vida, baseado na qualidade do cuidado materno. Métodos: Ao segundo dia de vida, genitoras Wistar e suas ninhadas foram divididas em dois grupos: intervenção, com redução do material para a confecção do ninho, ou controle. O comportamento materno foi observado do dia 1 ao dia 9 de vida. Na vida adulta, realizaram-se testes comportamentais e determinaram-se os níveis hipocampais de NO, através da mensuração de seus produtos de degradação. Resultados: Observou-se um maior comportamento do tipo ansioso no grupo intervenção, cujas genitoras apresentaram maior contato de baixa qualidade com seus filhotes. Nos machos, o cuidado materno de baixa qualidade correlacionou-se negativamente com o tempo no braço aberto e a frequência de mergulhos (r=-0,4;p=0,03) avaliados no labirinto em cruz elevado. O comportamento materno de lamber os filhotes correlacionou-se com a frequência de mergulhos em ambos os sexos (r=0,5;p<0,001). A quantidade de NO no hipocampo não diferiu entre os grupos. Conclusão: Uma maior atividade da nNOS não parece estar envolvida no comportamento ansioso observado neste modelo animal, no entanto a relação mãe-filhote, alterada por um ambiente neonatal adverso, teve impacto sobre o comportamento ansioso de forma sexo específica.

Background: Studies have shown that an adverse early life environment and the serotonergic system are related to the development of anxiety. This association could be mediated by neuronal nitric oxide synthase (nNOS), an enzyme that plays an important role in serotonin receptor functioning. Aim: To investigate the possible role of nNOS in the development of anxiety by measuring hippocampal nitric oxide (NO) in an animal model of neonatal stress, based on the quality of maternal care. Methods: On the second day of life, Wistar dams and their litters where divided in two groups: intervention, with limited access to nesting material, or control. Maternal behavior was observed from day 1 to 9 of life. In adult life, behavioral tests were performed and hippocampal NO levels were determined by measuring its degradation products. Results: There was more frequent anxiety-like behavior in the intervention group, whose dams showed low quality contact with their pups more often. In males, low quality maternal care was negatively correlated with time spent in open arms and frequency of head dips (r=-0.4; p=0.03) assessed using an elevated plus maze. Licking and grooming score was correlated with frequency of head dips in both sexes (r=0.5;p<0.001). Hippocampal NO levels were not different between groups (p=0.992). Conclusion: A higher nNOS activity does not seem to be involved in anxiety-like behavior observed in this animal model, however the relationship between dam and pup, modified by an adverse early life environment, had a different impact on anxiety behavior between sexes.

O impacto da transição demográfico-epidemiológica na saúde da criança e do adolescente do Brasil / The impact demographic and epidemiological transition in the health of children and adolescents in Brazil

Esse artigo tem como objetivo discutir os novos desafios no campo da saúde da criança e do adolescente como conseqüência do intenso processo de transição demográfica e epidemiológica brasileiro. Os aspectos relacionados à assistência, à pesquisa e ao ensino dos profissionais de saúde e, a presença de um novo perfil de saúde e doença, com seus novos mecanismos de causalidade são objetos de reflexão buscando alternativas para o enfrentamento e a superação das dificuldades advindas dessa situação. Portanto, esse artigo tenta desenvolver uma análise crítica desse novo cenário na perspectiva de oferecer as respostas pertinentes para as novas demandas em saúde das futuras gerações de brasileiros.

The objective of the present study was to discuss the new challenges of children and adolescent health care related the intensive process of demographic and epidemiological transition in Brazil. Aspects related to health care, research, and education of health professionals, as well as the presence of a new profile of health and disease, including its new mechanisms of causality, are analyzed with the purpose of offering alternatives to face and overcome the difficulties arising from this situation. Therefore, we tried to conduct a critical analysis of this new scenario in order to provide relevant solutions to the new health demands of the next Brazilian generations.

Association between Na⁺,K⁺-ATPase activity and the vulnerability/resilience to mood disorders induced by early life experience.

There is increasing evidence that early life events can influence neurodevelopment and later susceptibility to disease. Chronic variable stress (CVS) has been used as a model of depression. The objective of this study was to evaluate the interaction between early experience and vulnerability to chronic variable stress in adulthood, analyzing emotional, metabolic and neurochemical aspects related to depression. Pups were (1) handled (10 min/day) or (2) left undisturbed from day 1 to 10 after birth. When the animals reached adulthood, the groups were subdivided and the rats were submitted or not to CVS, which consisted of daily exposure to different stressors for 40 days, followed by a period of behavioral tasks, biochemical (plasma corticosterone and insulin sensitivity) and neurochemical (Na⁺,K⁺-ATPase activity in hippocampus, amygdala and parietal cortex) measurements. Neonatally-handled rats demonstrated shorter immobility times in the forced swimming test, independently of the stress condition. There was no difference concerning basal corticosterone or insulin sensitivity between the groups. Na⁺,K⁺-ATPase activity was decreased in hippocampus and increased in the amygdala of neonatally-handled rats. CVS decreased the enzyme activity in the three structures, mainly in the non-handled group. These findings suggest that early handling increases the ability to cope with chronic variable stress in adulthood, with animals showing less susceptibility to neurochemical features associated with depression, confirming the relevance of the precocious environment to vulnerability to psychiatric conditions in adulthood.

Long lasting sex-specific effects upon behavior and S100b levels after maternal separation and exposure to a model of post-traumatic stress disorder in rats.

This study was undertaken to verify if repeated long-term separation from dams would affect the development of parameters related to post-traumatic stress disorder (PTSD) after animals are subjected to inescapable shock when adults. Wistar rats were subjected to repeated maternal separation during post-natal days 1-10. When adults, rats from both sexes were submitted to a PTSD model consisting of exposure to inescapable footshock, followed by situational reminders. We observed long-lasting effects of both interventions. Exposure to shock increased fear conditioning. Anxiety-like behavior was increased and exploratory activity decreased by both treatments, and these effects were more robust in males. Additionally, basal corticosterone in plasma was decreased, paralleling effects observed in PTSD patients. Levels of S100B protein in serum and cerebrospinal fluid (CSF) were measured. Levels in serum correlated with the effects observed in anxiety-like behavior, increasing in males exposed to shock, and presenting no effect in females. S100B in CSF was increased in females submitted to maternal separation during the neonatal period. These results suggest that, in rats, an early stress experience such as maternal separation may aggravate some effects of exposure to a stressor during adult age, and that this effect is sex-specific. Additionally, data suggest that the increased S100B levels, observed in serum, have an extracerebral origin, possibly mediated by an increase in the noradrenergic tonus. Increased S100B in brain could be related to its neurotrophic actions.

Satiety assessment in neonatally handled rats.

We have previously demonstrated that neonatal handling increases sweet food ingestion. In the present study, we examined whether food intake, using different kinds of food, is altered in neonatally handled animals, with or without inducing satiety using a sucrose solution. Abdominal fat, glycemia and hormones linked to appetite including leptin, ghrelin and insulin were also measured. We tested palatable food consumption in the homecage to verify whether environmental cues could influence ingestion. Nests of Wistar rats were either (1) non-handled or (2) handled (10 min/day). Handling was performed on days 1-10 after birth. When adults, rats were habituated to sweet food (Froot Loops, Kellogg's) and to palatable fiber pellets (Fiber One), Nestlé). Sweet food consumption was increased in the neonatally handled group, when tested in the homecage, and also in the satiety experiment. These rats displayed a satiety curve when compared to the control group, which ate less but constantly. Handled rats exposed to a sucrose solution decreased sweet food ingestion, which did not occur in the control group. When exposed to a food with complex carbohydrates, these differences disappeared. There were no differences in body weight, abdominal fat or in glycemia, as well as no differences in plasma levels of insulin or leptin. However, ghrelin was decreased in neonatally handled rats. Neonatally handled rats demonstrated an increased consumption of sweet food, satiety responses to sucrose, as well as decreased levels of plasma ghrelin. It is possible that signaling mechanisms related to satiety, both peripherally and/or centrally may contribute to these behavioral findings.