Oral exposure to food grade titanium dioxide (E171) induces intestinal and behavioural alterations in adult mice but limited effects in young mice.

BACKGROUND: Food-grade titanium dioxide (E171), a white colourant widely used in ultra-processed food products, has been banned in the European Union. However, its usage is still permitted in medicines, and in several other countries. The estimated intake of E171 in children is higher than in adults, which led us to hypothesise that E171 induces differential effects depending on age, with adult mice being the most susceptible due to age, despite the lower dose. AIM: To evaluate the effects of oral administration of E171 on intestinal permeability, ileum, and colon histology, and how these effects impact anxious and depressive behaviour in young and adult mice of both sexes. METHODS: Young and adult mice of both sexes C57BL/6 mice received 10 mg/kgbw E171/3 times per week for 3 months. E171 was administered orally in water by pipetting, while control groups only received drinking water, then intestinal permeability, histology and animal behaviour were analysed. RESULTS: E171 showed an amorphous shape, primary particles sized below 1 µm and anatase crystalline structure. Oral administration of E171 disrupted the intestinal permeability in adult male and female mice, but no effects were observed in young mice of both sexes. E171 promoted ileal adenoma formation in half of the adult female population, moreover hyperplastic crypts, and hyperplastic goblet cells at histological level in adult mice of both sexes. The colon presented hyperplastic goblet cells, hyperchromatic nuclei, increased proliferation and DNA damage in adult mice of both sexes. The anxiety and depressive behaviour were only altered in adult mice treated with E171, but no changes were detected in young animals of both sexes. CONCLUSIONS: Adult mice displayed higher susceptibility in all parameters analysed in this study compared to young mice of both sexes.

Δ9-Tetrahydrocannabinol Treatment Modifies Insulin Secretion in Pancreatic Islets from Prediabetic Mice Under Hypercaloric Diet.

Background: The endocannabinoid system over-activation is associated with type-2 diabetes mellitus onset, involving physiological, metabolic, and genetic alterations in pancreatic islets. The use of Δ9-Tetrahydrocannabinol (THC) as treatment is still controversial since its effects and mechanisms on insulin secretion are unclear. The aim of this study was to evaluate the effects of THC treatment in pancreatic islets from prediabetic mice. Methods: Prediabetes was induced in mice by hypercaloric diet, and then treated with THC for 3 weeks. Blood glucose and body weight were determined, after behavior tests. Histological changes were evaluated in whole pancreas; in isolated islets we analyzed the effect of THC exposure in glucose-stimulated insulin secretion (GSIS), gene expression, intracellular cyclic adenosine monophosphate (cAMP), and cytosolic calcium changes. Results: THC treatment in prediabetic mice enhanced anxiety and antidepressive behavior without changes in food ingestion, decreased oral-glucose tolerance test, plasma insulin and weight, with small alterations on pancreatic histology. In isolated islets from healthy mice THC increased GSIS, cAMP, and CB1 receptor (CB1r) expression, meanwhile calcium release was diminished. Small changes were observed in islets from prediabetic mice. Conclusions: THC treatment improves some clinical parameters in prediabetic mice, however, in isolated islets, modifies GSIS, intracellular calcium and gene expression, suggesting specific effects related to diabetes evolution.

El consumo de bebidas dulces incrementa diferencialmente la expresión de Oxitocina en núcleos hipotalámicos / The consumption of sweet drinks differentially increases the expression of oxytocin in hypothalamic nuclei

Resumen Se reconoce la participación de la oxitocina en el control de la alimentación, pero su mecanismo de acción no se ha establecido totalmente. Por tanto, el objetivo de esta investigación fue evaluar el efecto del acceso intermitente a una solución de sacarosa, sobre la expresión de las neuronas del núcleo paraventricular (PVN) y del núcleo supraótico (SON) que producen oxitocina (Oxt), y caracterizar la microestructura de la conducta de beber en ratas saciadas. Se tuvieron tres grupos de ratas macho Wistar saciadas, y en la primera hora al inicio del periodo de luz, el grupo Control tuvo agua, el grupo Restringido 5g de una solución de sacarosa al 20% y el grupo Ad libitum acceso libre a la solución de sacarosa. Los sujetos incrementaron el consumo de la solución de sacarosa a pesar de estar saciados; debido a la interrupción del estado de saciedad y la demora de la satisfacción. La actividad de las neuronas de Oxt se incrementó en ambos núcleos, en el grupo Restringido la mayor expresión se observó en el SON y en el grupo Ad libitum en el PVN. No se encontró correlación entre la cantidad de bebida ingerida y la actividad de las neuronas Oxt.

Abstract The role of oxytocin in feeding control is recognized, but its mechanism of action has not been fully established. Therefore, the aim of this research was to evaluate the effect of intermittent access to a sucrose solution on the expression of paraventricular nucleus (PVN) and supraotic nucleus (SON) neurons that produce oxytocin (Oxt), and to characterize the microstructure of drinking behavior in satiated rats. Three groups of male Wistar rats satiated were used, and in the first hour at the beginning of the light period, a Control group had water, a Restricted group 5g of a 20% sucrose solution and Ad libitum group with free access to sucrose solution. The experimental subjects increased the consumption of the sucrose solution despite being satiated, due to the interruption of the state of satiety and the delay of the satiation process. Oxt neurons increased their activity in both nuclei, in the Restricted group the highest expression was observed in the SON and in the Ad libitum group in the PVN. No correlation was found between the amount of drink ingested and the activity of Oxt neurons.

The central blockade of the dopamine DR4 receptor decreases sucrose consumption by modifying the microstructure of drinking behavior in male rats.

Sugar solutions promote hedonic feeding and increase the risk of obesity and binge-type behavior. In rodents, ingestion of sugar solutions enhances dopamine release to mesolimbic regions, suggesting changes in hedonic intake and brain reward processes. Moreover, dopaminergic D2R/D3R receptors contribute to the hedonic intake of palatable solutions. Although the experimental evidence indicate that the dopaminergic D4 receptor (D4R) modulates feeding at homeostatic levels, it is currently unknown whether D4R also regulate the hedonic intake of sugar solutions. In this study, we evaluated the effect of the central blockade of D4R on the consumption of a 20% sucrose solution, the drinking microstructure parameters, and levels of locomotor activity in sated rats. In the first experiment, male Wistar rats were daily exposed to a 20% sugar solution in the first hour of the light phase of the light:dark cycle. On day 10, rats received i.c.v injections of the D4R antagonist, L-745870 (0, 1 or 2 µg/5 µl) and sucrose consumption and drinking microstructure parameters (latency to start drinking, bouts, drinking duration, bout size, inter-bout interval, time in activity and time in resting) were evaluated. In the second experiment, rats were trained to receive the 20% sucrose solution as described in experiment 1. On day 10, after the 1 h of sucrose access, the rats were placed in the open field for 5-min (habituation phase). Then, rats received i.c.v injections of L-745870 (0, 1 or 2 µg/ 5 µl), and were placed again in the open-field test for 10-min (pharmacological phase). The number or crosses trough squares and number of rears were scored for both the habituation and pharmacological phase. Here we found that administration of L-745870 decreased the consumption of sucrose in a dose-depended manner. Moreover, L-745870-treated rats displayed microstructural changes, including greater number of bouts and reduced drinking duration, bout size and inter-bout intervals. Furthermore, the number of crosses and number of rears in the open field test remained unchanged for habituation and pharmacological phase. Finally, present findings suggest that D4R modulates the consumption of sugar solutions by alteration of hedonic responses, but the contribution of homeostatic systems is discussed. These results open perspectives for the potential use of the D4R antagonists for treating obesity or binge-eating behavior.

Food-grade titanium dioxide (E171) differentially affects satiation in mice fed a regular or a high fat diet.

Food-grade titanium dioxide (E171) is a widely used food additive and the toxicity after oral consumption is still under research, although it has been already banned in some countries. The consumption of this additive occurs mainly through ultra-processed food products which also contain high amounts of fat. High fat diets (HFD) impair the physiological system controlling satiation and satiety, which are responsible for control of food intake and energy status. The impact of E171 on animal behavior has been poorly explored and here we hypothesize that E171 could worsen the effects on feeding behavior induced by HFD. Therefore, we aimed to evaluate the effects of E171 on the feeding pattern and the behavioral satiety sequence (BSS) of mice fed with a regular diet (RD) or a HFD after 1 and 16 weeks of exposure. The results showed that RD + E171 increased food intake and feeding time, but the prototypical structure of the BSS pattern (feeding→ grooming-activity → resting), was preserved. Conversely, food consumption was not altered in HFD + E171, but the BSS pattern was disrupted as the animals prolonged resting time and spent less time being active. Our findings suggest that E171 delayed the onset of satiation in mice fed with RD but induced the opposite effect in mice fed with HFD.

Food-grade titanium dioxide decreases hematocrit and hemoglobin and increases compulsive-like behavior in male mice.

Food-grade titanium dioxide (E171) is widely used as a food additive, and it is known that after oral consumption, E171 is translocated into the bloodstream reaching the highest titanium level at 6 h. E171 is accumulated in some organs triggering toxicity, but the effects on the blood parameters after oral consumption have been less studied. Recently, evidence shows that oral exposure to E171 induces behavioral signs of anxiety and depression. The relation between blood alterations and psychiatric disorders has been previously demonstrated. However, the oral exposure to E171 effects on alterations in blood parameters and effects linked to alterations in animal behavior has not been explored. In this short communication, we aimed to investigate the effects of E171 on specific blood parameters (hematocrit, hemoglobin, number of erythrocytes, and leukocytes) and anxiety and compulsive-like behavior in males and females orally exposed to ~5 mg/kg for 4 weeks. The results showed that E171 decreased hematocrit and hemoglobin in male but not in female mice while leukocyte and erythrocyte count remained unaltered. Oral consumption of E171 decreased the levels of anxiety-like behavior in females but not in male mice, while compulsive-like behavior was increased in both male and female mice.

Stimulation of melanocortin-3 receptors accelerates the satiation process and increases the α-MSH expression in high-fat diet-fed rats.

The knowledge about the role of MC3 receptors (MC3r) in the regulation of feeding behavior is limited. The present study was conducted to determine whether MC3r mediates the hypophagic effects of the melanocortins under conditions of positive energy balance. Male Wistar rats were fed with a high-fat diet (HFD) for 15 days and on day 16 the animals received an intracerebroventricular injection of the following treatments: Vehicle, D-Trp8-γ-melanocyte-stimulating hormone (MSH; MC3r agonist), SHU9119 (MC3r/MC4r antagonist), or D-Trp8-γ-MSH+SHU9119. Food intake was measured and the behavioral satiety sequence (BSS) analysis was carried out during the first hour of the dark phase. The c-Fos and α-MSH immunoreactivity in the arcuate nucleus (ARC) was evaluated 60 min later the onset of food intake. The results indicated that D-Trp8-γ-MSH decreased the ingestion of the HFD and this effect is associated with the early development of the satiation process Moreover, the D-Trp8-γ-MSH increased the accumulation of the α-MSH in the ARC and the c-Fos activity in the PVN. The antagonist SHU9119 partially prevented the D-Trp8-γ-MSH-induced hypophagia. Moreover, behavioral analysis suggests that central activation of MC3r accelerated the cessation of feeding in conditions of positive energy balance; the possible role of MC4r is discussed. Present data indicate that central stimulation of MC3r prevented the overconsumption of the HFD without affecting the natural satiation process, suggesting a potential use of MC3r for the treatment of eating disorders that are stimulated by hypercaloric diets. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Melanocortinas y su participación en el desarrollo de la obesidad

Resumen La obesidad y el sobrepeso son problemas de origen multifactorial, la interacción de los factores ambientales, genéticos y conductuales parecen ser la clave en el desarrollo de esta patología. Los receptores MC3 (rMC3) y MC4 (rMC4), participan en la regulación del balance energético (consumo de alimento, gasto energético y peso corporal), su mutación genética está asociada con el incremento de la adiposidad y el desarrollo de la obesidad. El conocimiento que se tiene de estos receptores y su función aún es limitado, principalmente con respecto a los rMC3 y, aunque los rMC4 se están posicionando como un blanco terapéutico potencial para el tratamiento de la obesidad, se requiere de mayor investigación clínica. Por lo tanto, el propósito de este documento es presentar evidencia de la participación de los receptores MC3 y MC4 en el desarrollo de la obesidad, a través de los resultados encontrados en algunas investigaciones, tanto en modelos animales como en humanos.

Abstract Obesity and overweight are problems of multifactorial origin, the interaction of environmental, genetic, and behavioral factors seems to be the key in the development of this pathology. The MC3 (rMC3) and MC4 (rMC4) receptors participate in the regulation of energy balance (food intake, energy expenditure and body weight), their genetic mutation is associated with increased adiposity and the development of obesity. Knowledge of these receptors and their function is still limited, mainly with respect to rMC3 and, although rMC4 are positioning themselves as a potential therapeutic target for the treatment of obesity, further clinical research is required. Therefore, the purpose of this document is to present the participation of MC3 and MC4 receptors in the development of obesity, through the results found in some investigations, both in animal and in human´s models.

Food additives containing nanoparticles induce gastrotoxicity, hepatotoxicity and alterations in animal behavior: The unknown role of oxidative stress.

Food additives such as titanium dioxide (E171), iron oxides and hydroxides (E172), silver (E174), and gold (E175) are highly used as colorants while silicon dioxide (E551) is generally used as anticaking in ultra-processed foodstuff highly used in the Western diets. These additives contain nanosized particles (1-100 nm) and there is a rising concern since these nanoparticles could exert major adverse effects due to they are not metabolized but are accumulated in several organs. Here, we analyze the evidence of gastrotoxicity, hepatotoxicity and the impact of microbiota on gut-brain and gut-liver axis induced by E171, E172, E174, E175 and E551 and their non-food grade nanosized counterparts after oral consumption. Although, no studies using these food additives have been performed to evaluate neurotoxicity or alterations in animal behavior, their non-food grade nanosized counterparts have been associated with stress, depression, cognitive and eating disorders as signs of animal behavior alterations. We identified that these food additives induce gastrotoxicity, hepatotoxicity and alterations in gut microbiota and most evidence points out oxidative stress as the main mechanism of toxicity, however, the role of oxidative stress as the main mechanism needs to be explored further.

Food-grade titanium dioxide (E171) induces anxiety, adenomas in colon and goblet cells hyperplasia in a regular diet model and microvesicular steatosis in a high fat diet model.

Food-grade titanium dioxide (E171) is a white additive widely used in solid and liquid food products. There is still debate about E171 toxic effects after oral consumption since this additive is deposited in colon, liver, spleen, testis and brain. The consumption of E171 commonly occurs with Western diets that are characterized by a high fat content. Thus, E171 could worsen adverse effects associated with a high fat diet (HFD) such as anxiety, colon diseases and testicular damage. We aimed to evaluate the effects of E171 on anxiety-like behavior, colon, liver and testis and to analyze if the administration of a HFD could exacerbate adverse effects. E171 was administered at ~5 mg/kgbw by drinking water for 16 weeks and mice were fed with a Regular Diet or a HFD. E171 promoted anxiety, induced adenomas in colon, goblet cells hypertrophy and hyperplasia and mucins overexpression, but had no toxic effects on testicular tissue or spermatozoa in regular diet fed-mice. Additionally, E171 promoted microvesicular steatosis in liver in HFD fed-mice and the only HFD administration decreased the spermatozoa concentration and motility. In conclusion, E171 administration increases the number of adenomas in colon, induces hypertrophy and hyperplasia in goblet cells and microvesicular steatosis.

Effects of a high-fat diet on behavioral eating patterns / Efeitos de uma dieta com alto conteúdo de gordura sobre padrões comportamentais alimentares / Efectos de una dieta con alto contenido de grasas sobre patrones conductuales alimentarios

Abstract Excessive consumption of high-fat food has been associated with increased prevalence of obesity. The physiological and metabolic effects of high-fat diets have been extensively studied. Nevertheless, the behavioral mechanisms associated with the development of obesity induced by consumption of these diets has been less explored. Therefore, the aim of the present study was to characterize the changes in the behavioral feeding patterns produced by the consumption of a high-fat diet during 10 days. Male Wistar rats with free access to food were assigned to one of two groups, and for 10 days, they had access to a high-fat diet (45 % calories from fat) or to a standard diet. Detailed analysis of feeding behavior was performed on days 1, 5 and 10 at the beginning of the dark period. The results showed that subjects exposed to the high-fat diet accumulated more body fat and showed increased feeding efficiency, in absence of excessive body weight increase or alterations in the behavioral satiety sequence pattern. These findings suggest that exposure to high-fat diets may produce behavioral changes before excessive gain of body weight occurs, primarily affecting control mechanisms of feeding efficiency.

Resumo O consumo excessivo de alimentos com alto conteúdo de gordura tem sido associado com o aumento da obesidade. Os efeitos fisiológicos e metabólicos do consumo de dietas altas em gordura têm sido estudados extensamente, contudo os mecanismos comportamentais relacionados com o desenvolvimento da obesidade pelo consumo dessas dietas têm-se explorado em menor medida. Portanto, o objetivo deste estudo foi caracterizar as mudanças nos padrões comportamentais da alimentação produzidas pelo consumo de uma dieta alta em gordura durante dez dias. Utilizaram-se ratos de laboratório macho Wistar, com acesso livre ao alimento, designados a um de dois grupos, e durante dez dias estiveram sob uma dieta alta em gordura (45 % de calorias provenientes de gorduras) ou uma dieta padrão de laboratório. Nos dias 1, 5 e 10, realizou-se uma análise detalhada do comportamento alimentar ao início do período de escuridão. Os resultados mostraram que os sujeitos expostos à dieta alta em gordura acumularam mais gordura corporal e tiveram maior eficiência da alimentação do que o outro grupo, sem aumento do peso corporal nem alterações do padrão da sequência de saciedade comportamental. Isso sugere que a exposição a dietas com alto conteúdo de gordura pode produzir mudanças comportamentais antes de apresentar um ganho de peso excessivo, o que afeta principalmente os mecanismos de controle de eficiência alimentar.

Resumen El consumo excesivo de alimentos con alto contenido de grasas se ha asociado con el incremento de la obesidad. Los efectos fisiológicos y metabólicos del consumo de dietas altas en grasa han sido estudiados extensamente; sin embargo, los mecanismos conductuales asociados al desarrollo de la obesidad por el consumo de estas dietas se han explorado en menor medida. Por tanto, el objetivo del presente estudio fue caracterizar los cambios en los patrones conductuales de la alimentación producidos por el consumo de una dieta alta en grasas durante diez días. Se utilizaron ratas macho Wistar con acceso libre al alimento, asignadas a uno de dos grupos, y durante diez días estuvieron bajo una dieta alta en grasa (45 % de calorías provenientes de grasas) o una dieta estándar de laboratorio. En los días 1, 5 y 10 se realizó un análisis detallado de la conducta alimentaria al inicio del periodo de oscuridad. Los resultados mostraron que los sujetos expuestos a la dieta alta en grasa acumularon más grasa corporal y tuvieron mayor eficiencia de la alimentación que el otro grupo, sin incremento del peso corporal ni alteraciones del patrón típico de la secuencia de saciedad conductual. Esto sugiere que la exposición a dietas con alto contenido de grasas puede producir cambios conductuales antes de que se presente una ganancia de peso excesivo, lo que afecta principalmente los mecanismos de control de eficiencia alimentaria.