Biochemical and molecular evaluation of oxidative stress and mitochondrial damage in fruit fly exposed to carmoisine.

BACKGROUND: In today's world, appearance is an important factor in almost all areas of our lives. Therefore, it has become common to use dyes to color foods to make them look appetizing and visually appealing. However, food additives have negative effects on biochemical processes in cells at both high and low doses. METHODS AND RESULTS: This study investigated the effect of carmoisine, a commonly used food coloring, on oxidative stress and damage parameters in Drosophila melanogaster in terms of both enzymatic and gene expression. The change in mitochondrial DNA copy number (mtDNA-CN), a marker of oxidative stress, was also examined. When the data obtained were analyzed, it was observed that carmoisine caused a significant decrease in GSH levels depending on the increase in dose. SOD, CAT, GPx, and AChE enzyme activities and gene expression levels were also found to be significantly decreased. All groups also showed a significant decrease in mtDNA-CN. The effect of carmoisine on Drosophila melanogaster morphology was also investigated in our study. However, no significant change was observed in terms of morphological development in any group. CONCLUSIONS: When all the findings were evaluated together, it was observed that carmoisin triggered oxidative stress and these effects became more risky at high doses. Therefore, we believe that the consumer should be made more aware of the side effects of azo dyes in food and that the type and concentration of each substance added to food should be specified.

Tartrazine Modifies the Activity of DNMT and HDAC Genes-Is This a Link between Cancer and Neurological Disorders?

In recent years, artificial additives, especially synthetic food colorants, were found to demonstrate wider properties compared to their natural equivalents; however, their health impact is still not totally mapped. Our study aimed to determine the long-term (30 and 90 days) exposure effect of one of the commonly used artificial food colorants, tartrazine, on NMRI mice. The applied dose of tartrazine referred to the human equivalent dose for acceptable daily intake (ADI). Further, we evaluated its impact on the transcription of a range of epigenetic effectors, members of the DNA methyltransferase (DNMT) as well as histone deacetylase (HDAC) families. Following the exposure, organ biopsies were collected from the lungs, kidneys, liver, and spleen, and the gene expression levels were determined by real-time quantitative reverse transcription PCR (RT-qPCR). Our results demonstrated significant upregulation of genes in the tested organs in various patterns followed by the intake of tartrazine on ADI. Since DNMT and HDAC genes are involved in different steps of carcinogenesis, have roles in the development of neurological disorders and the effect of dose of everyday exposure is rarely studied, further investigation is warranted to study these possible associations.

Food Color Additives in Hazardous Consequences of Human Health: An Overview.

Food color additives are used to make food more appetizing. The United States Food and Drug Administration (FDA) permitted nine artificial colorings in foods, drugs, and cosmetics, whereas the European Union (EU) approved five artificial colors (E-104, 122, 124, 131, and 142) for food. However, these synthetic coloring materials raise various health hazards. The present review aimed to summarize the toxic effects of these coloring food additives on the brain, liver, kidney, lungs, urinary bladder, and thyroid gland. In this respect, we aimed to highlight the scientific evidence and the crucial need to assess potential health hazards of all colors used in food on human and nonhuman biota for better scrutiny. Blue 1 causes kidney tumor in mice, and there is evidence of death due to ingestion through a feeding tube. Blue 2 and Citrus Red 2 cause brain and urinary bladder tumors, respectively, whereas other coloring additives may cause different types of cancers and numerous adverse health effects. In light of this, this review focuses on the different possible adverse health effects caused by these food coloring additives, and possible ways to mitigate or avoid the damage they may cause. We hope that the data collected from in vitro or in vivo studies and from clinical investigations related to the possible health hazards of food color additives will be helpful to both researchers and the food industry in the future.

IFN-γ+ cytotoxic CD4+ T lymphocytes are involved in the pathogenesis of colitis induced by IL-23 and the food colorant Red 40.

The food colorant Red 40 is an environmental risk factor for colitis development in mice with increased expression of interleukin (IL)-23. This immune response is mediated by CD4+ T cells, but mechanistic insights into how these CD4+ T cells trigger and perpetuate colitis have remained elusive. Here, using single-cell transcriptomic analysis, we found that several CD4+ T-cell subsets are present in the intestines of colitic mice, including an interferon (IFN)-γ-producing subset. In vivo challenge of primed mice with Red 40 promoted rapid activation of CD4+ T cells and caused marked intestinal epithelial cell (IEC) apoptosis that was attenuated by depletion of CD4+ cells and blockade of IFN-γ. Ex vivo experiments showed that intestinal CD4+ T cells from colitic mice directly promoted apoptosis of IECs and intestinal enteroids. CD4+ T cell-mediated cytotoxicity was contact-dependent and required FasL, which promoted caspase-dependent cell death in target IECs. Genetic ablation of IFN-γ constrained IL-23- and Red 40-induced colitis development, and blockade of IFN-γ inhibited epithelial cell death in vivo. These results advance the understanding of the mechanisms regulating colitis development caused by IL-23 and food colorants and identify IFN-γ+ cytotoxic CD4+ T cells as a new potential therapeutic target for colitis.

Artificial food coloring affects EEG power and ADHD symptoms in college students with ADHD: a pilot study.

Objectives: Removing artificial food coloring (AFC) is a common dietary intervention for children with Attention-Deficit/Hyperactivity Disorder (ADHD), but has not been tested in young adults. This pilot study examined the effects of AFC on ADHD symptoms and electroencephalography (EEG) in college students with and without ADHD.Methods: At baseline, control and ADHD participants completed the Adult ADHD Self-Report Scale (ASRS), simple and complex attention measures, and resting-state EEG recordings. ADHD participants (n = 18) and a subset of controls (extended control group or EC, n = 11) avoided AFC in their diet for 2 weeks and then were randomized to a double-blind, placebo-controlled crossover challenge. Subjects received either 225 mg AFC disguised in chocolate cookies or placebo chocolate cookies for 3 days each week, with testing on the third day each week. Baseline comparisons were made using Student's t-test or Wilcoxon rank sum tests and challenge period analyses were run using General Linear Modeling.Results: The ADHD group had significantly greater scores on the ASRS (p < 0.001), confirming a symptom differential between groups; however, there were no differences in attentional measures or EEG at baseline. The AFC challenge resulted in an increase in posterior mean gamma power (p = 0.05), a decrease in posterior relative alpha power (p = 0.04), and a marginal increase in inattentive symptoms (p = 0.08) in the ADHD group. There were no effects of AFC in the EC group.Discussion: This study indicates that AFC exposure may affect brainwave activity and ADHD symptoms in college students with ADHD. Larger studies are needed to confirm these findings.

Carmines (E120) in coloured yoghurts: a case-study contribution for human risk assessment.

Carmines (E120) are used worldwide as natural food colouring agents of animal origin, with a widespread application, including yoghurts. Despite being considered safe for human health, carmines are known to cause allergic reactions. Our goal was to evaluate the presence of carmines in different yoghurts with a label declaration of E120, purchased in Portugal, and, for the first time, to assess the human risk. The analytical methodology, recommended by JECFA, was based on acid hydrolysis of the samples followed by spectrophotometric UV-Vis analysis at 494 nm. This methodology allowed for a limit of quantification (LOQ) of 39.0 mg/kg and recovery rates higher than 97.7%. All the samples had carmines at levels above the LOQ, ranging between 43.8 and 193.8 mg/kg, with an average of 125.2 ± 34.5 mg/kg. In total, 8 (26.7%) samples exceeded the European Union (EU) maximum permitted level (MPL) established for carmines in this foodstuff, 150 mg/kg. Solid yoghurts presented higher average levels, 137.2 mg/kg, when compared to liquid samples, 107.2 mg/kg, with a significant statistical difference (p= 0.0236) being observed. No significant statistical difference was observed between white and private labels, whose average levels were very similar, 125.4 vs 125 mg/kg, respectively. Although some samples were above the allowable values, the estimated daily intake (EDI), designed for the different scenarios of different yoghurt types, did not exceed the established ADI, 5 mg/kg bw/day. According to the obtained results, carmine ingestion through the consumption of yoghurt poses low risk to the Portuguese consumers. However, children were the most vulnerable population group with a calculated risk value of up to 10% considering the mean content scenario. These first findings point out the need to reinforce surveillance programmes and monitoring studies, contributing to an increased awareness regarding carmine exposure, however it must be emphasised that yoghurt has evident nutritional benefits depending on a healthful consumer choice.

National clinical guideline for assessment and treatment of ADHD in children and young people: quick guide

Professional observers are independent persons who have in-depth knowledge of both children's normal behaviour and development and of ADHD, and who take a multidisciplinary approach to ADHD in their daily work. The observation must be geared to the specific problem with the child in question

Food colorants metabolized by commensal bacteria promote colitis in mice with dysregulated expression of interleukin-23.

Both genetic predisposition and environmental factors appear to play a role in inflammatory bowel disease (IBD) development. Genetic studies in humans have linked the interleukin (IL)-23 signaling pathway with IBD, but the environmental factors contributing to disease have remained elusive. Here, we show that the azo dyes Red 40 and Yellow 6, the most abundant food colorants in the world, can trigger an IBD-like colitis in mice conditionally expressing IL-23, or in two additional animal models in which IL-23 expression was augmented. Increased IL-23 expression led to generation of activated CD4+ T cells that expressed interferon-γ and transferred disease to mice exposed to Red 40. Colitis induction was dependent on the commensal microbiota promoting the azo reduction of Red 40 and generation of a metabolite, 1-amino-2-naphthol-6-sulfonate sodium salt. Together these findings suggest that specific food colorants represent novel risk factors for development of colitis in mice with increased IL-23 signaling.

Integration of evidence to evaluate the potential for neurobehavioral effects following exposure to USFDA-approved food colors.

California's Office of Environmental Health Hazard Assessment was tasked with conducting risk assessments for United States Food and Drug Administration-approved food dyes relative to neurobehavioral concerns. The purpose of this assessment was to evaluate the evidence for neurodevelopment effects based on three streams of evidence: 1) studies identified by OEHHA for consideration in a quantitative risk assessment; 2) studies relevant to understanding mechanisms of neurobehavioral effects; 3) an in silico assessment of the bioavailability of USFDA-approved food dyes. The results indicate a lack of adequate or consistent evidence of neurological effects, supported by a lack of bioavailability and brain penetration predicted by the in silico assessment. Further, the mechanistic evidence supports a lack of activity from in vitro neurotransmitter assays, and a lack of evidence to support molecular initiating events or key events in adverse outcome pathways associated with neurodevelopmental effects, supporting a lack of biological plausibility for neurobehavioral effects following food exposures to colors. These conclusions are consistent with other authoritative bodies, such as JECFA and EFSA, that have determined (i) other effects are more appropriate for estimating acceptable daily intakes and (ii) evidence from the neurobehavioral studies lack the strength to be relied upon for quantitative risk assessment.

Prenatal exposure to artificial food colorings alters NMDA receptor subunit concentrations in rat hippocampus.

Exposure to artificial food color additives (AFCAs) has been implicated in the etiology of certain childhood hyperactivity and learning disabilities. N-methyl-D-aspartate receptors and alpha-7 nicotinic acetylcholine receptor (α7 nAChR) are involved in learning and memory. We administered a mixture of AFCAs (erythrosine, ponceau 4R, allura red AC, sunset yellow FCF, tartrazine, amaranth, brilliant blue, azorubine, and indigotine) to female rats during gestation to investigate the effects of prenatal exposure to AFCAs on neurobehavior, spatial learning, and memory in their offspring. We also investigated whether AFCAs modulate NR2A, NR2B, and α7 nAChR protein levels in their offsprings' hippocampi. Although spatial learning and memory were not altered, the offspring of rats exposed to AFCAs exhibited decreased motivation and increased despair-related behavior. NR2A and NR2B protein levels were significantly reduced in female offspring in the experimental group (p < 0.05), whereas α7 nAChR level was not significantly altered. Our results suggest that prenatal exposure to AFCAs may lead to sex-dependent alterations in glutamatergic signaling which may continue into adolescence.

Edible additive effects on zebrafish cardiovascular functionality with hydrodynamic assessment.

Food coloring is often used as a coloring agent in foods, medicines and cosmetics, and it was reported to have certain carcinogenic and mutagenic effects in living organisms. Investigation of physiological parameters using zebrafish is a promising methodology to understand disease biology and drug toxicity for various drug discovery on humans. Zebrafish (Danio rerio) is a well-acknowledged model organism with combining assets such as body transparency, small size, low cost of cultivation, and high genetic homology with humans and is used as a specimen tool for the in-vivo throughput screening approach. In addition, recent advances in microfluidics show a promising alternative for zebrafish manipulation in terms of drug administration and extensive imaging capability. This pilot work highlighted the design and development of a microfluidic detection platform for zebrafish larvae through investigating the effects of food coloring on cardiovascular functionality and pectoral fin swing ability. The zebrafish embryos were exposed to the Cochineal Red and Brilliant Blue FCF pigment solution in a concentration of (0.02‰, 0.2‰) cultured in the laboratory from the embryo stage to hatching and development until 9 days post fertilization (d.p.f.). In addition, zebrafish swimming behaviors in terms of pectoral fin beating towards the toxicity screening were further studied by visualizing the induced flow field. It was evidenced that Cochineal Red pigment at a concentration of 0.2‰ not only significantly affected the zebrafish pectoral fin swing behavior, but also significantly increased the heart rate of juvenile fish. The higher concentration of Brilliant Blue FCF pigment (0.2%) increased heart rate during early embryonic stages of zebrafish. However, zebrafish exposed to food coloring did not show any significant changes in cardiac output. The applications of this proposed platform can be further extended towards observing the neurobiological/hydrodynamic behaviors of zebrafish larvae for practical applications in drug tests.

Adverse reactions to food additives in children: A retrospective study and a prospective survey.

BACKGROUND: Allergic reactions to food additives are often suspected by families. The aim of this study was to describe oral food challenge (OFC) outcomes in a pediatric cohort with a suspected diagnosis of allergy to food additives (food dyes or sodium benzoate). METHODS: All patients who underwent an open OFC to carmine red, cochineal red, erythrosine, patent blue V, tartrazine, yellow sunset S, and/or sodium benzoate were included. A survey was sent to families after testing to evaluate whether the OFC results had altered feeding behaviors with food additives. RESULTS: Twenty-three patients were included. The main suspected food was candy (n=11/23; 48%). Only one OFC out of 45 was formally positive for the carmine and cochineal red. Subsequently, most OFCs were negative (44/45; 97.8%). Despite the negativity of the challenge, four families out of 14 reported occurrences of supposed allergic reactions to food additives and six out of 15 continued to completely avoid the additive of concern in their children's diet. CONCLUSIONS: Allergies to food additives remain rare. Even if an IgE-mediated allergy was excluded with a negative OFC, families remained suspicious about ready-made products. Health professionals and parents should be reassured about the low risk of food dye intolerance or allergies.

Food Color and Autism: A Meta-Analysis.

Autism has been increasing dramatically since its description by Leo Kanner in 1943. The Centers for Disease Control and Prevention (CDC) in 2018 has identified 1 in 59 children (1 in 37 boys and 1 in 151 girls) has autism spectrum disorder (ASD). Autistic spectrum disorders and ADHD are complex conditions in which nutritional and environmental factors play major roles. It is important to understand how food can have an impact on their current and future health. Appealing food colors stimulate the consumption of different food products. Since 2011, it is evident that dyes are linked to harmful effects in children. Artificial dyes have neurotoxic chemicals that aggravate mental health problems. Many families with autistic children avoid food dyes in their diet in order to avoid behavioral issues. A study reported that there is a correlation between yellow dye and sleep disturbance. Food colors Blue 1 and 2, Green 3, Red 3, Yellow 5 and 6, Citrus Red 2, and Red 40 can trigger many behaviors in most kids. Artificial food color usually contains petroleum and is manufactured in a chemical process that includes formaldehyde, aniline, hydroxides, and sulfuric acids. Most impurities in the food color are in the form of salts or acids. Sometimes lead, arsenic, and mercury may be present as impurities. The U.S. FDA is yet to study the effects of synthetic dyes on behavior in children. A study conducted at Southampton University in England found a link between food dyes and hyperactive behavior in children. The research does not prove that food coloring actually causes autism spectrum disorder, but there seems to be a link. This chapter attempts to provide a broad review of the available literature on food color and the epidemiology, etiology, prevention, and treatment of autistic spectrum disorder.

Effects of Titanium Dioxide Nanoparticles Exposure on Human Health-a Review.

Recently, an increased interest in nanotechnology applications can be observed in various fields (medicine, materials science, pharmacy, environmental protection, agriculture etc.). Due to an increasing scope of applications, the exposure of humans to nanoparticles (NPs) is inevitable. A number of studies revealed that after inhalation or oral exposure, NPs accumulate in, among other places, the lungs, alimentary tract, liver, heart, spleen, kidneys and cardiac muscle. In addition, they disturb glucose and lipid homeostasis in mice and rats. In a wide group of nanoparticles currently used on an industrial scale, titanium dioxide nanoparticles-TiO2 NPs-are particularly popular. Due to their white colour, TiO2 NPs are commonly used as a food additive (E 171). The possible risk to health after consuming food containing nanoparticles has been poorly explored but it is supposed that the toxicity of nanoparticles depends on their size, morphology, rate of migration and amount consumed. Scientific databases inform that TiO2 NPs can induce inflammation due to oxidative stress. They can also have a genotoxic effect leading to, among others, apoptosis or chromosomal instability. This paper gives a review of previous studies concerning the effects of exposure to TiO2 NPs on a living organism (human, animal). This information is necessary in order to demonstrate potential toxicity of inorganic nanoparticles on human health.

Rhodamine B in spices determined by a sensitive UPLC-MS/MS method.

Rhodamine B (RhB) is a banned food additive and has been classified as illegal colourant. Therefore, the risk of RhB contamination should be strictly monitored. In this study, a sensitive UPLC-MS/MS method was applied to monitor RhB in 292 various spices such as chilli, pepper and tomato products. The results showed 22.7% of chilli powder samples, 18.5% of pepper powder samples, 11.1% of chilli oil samples and 9.1% of pepper oil samples were contaminated with RhB. Chilli powder contained RhB up to 44,935 µg/kg with an average of 743 µg/kg, pepper powder up to 65.9 µg/kg with an average of 4.1 µg/kg, chilli oil up to 14.6 µg/kg with an average of 1.0 µg/kg and pepper oil up to 1.1 µg/kg with an average of 0.2 µg/kg, respectively. Considering the common consumption of chilli products and pepper products by so many consumers, RhB exposure is significant and should be decreased.

Corantes alimentares presentes em alimentos ultraprocessados consumidos por universitários / Colorantes alimentarios presentes en alimentos ultraprocesados consumidos por universitários / Food dyes present in ultra-processed foods consumed by university students

Objective: to describe the food dyes present in ultra-processed foods consumed by 273 students of the Nutrition Course at a public university of Rio de Janeiro. Methods: It was characterized the socio-demographic and health profile using semi-structured questionnaire. Consumption of ultra processed food was conducted by the Food Frequency Questionnaire (gelatine, stuffed cookies, candies and bubble gum, soda, powdered drink mix, industrialized juice, bouillon cube and instant noodles), and the food dyes were identified in the products’ label. Results: The consumption of products containing food dyes such as gelatines, candies and bubble gum, soda, and bouillon cube was above 80%. The candy and bubble gum, and bouillon cube consumption were almost daily, being 56.9% and 54.1% respectively. Conclusion: 14 food dyes food were identified on the labels of ultra-processed food. The artificial dyes caramel III and IV, bourdeaux S, sunset yellow and tartrazine, and the natural colorants annatto and carmine were highlighted.

Objetivo: descrever os corantes alimentares presentes nos alimentos ultraprocessados consumidos por 273 graduandos de uma universidade pública do Rio de Janeiro. Métodos: Foi caracterizado o perfil sociodemográfico e de saúde a partir de questionário semiestruturado. Consumo de alimentos ultraprocessados foi obtido através do Questionário de Frequência Alimentar (gelatinas, biscoitos recheados, balas e chicletes, refrigerantes, preparados sólidos para refresco, sucos industrializados, temperos prontos e macarrão instantâneo) e os corantes foram identificados nos rótulos dos produtos. Resultados: O consumo de produtos contendo corantes como gelatinas, balas e chicletes, refrigerantes e sucos industrializados foi acima de 80%. Balas e chicletes e temperos prontos tiveram consumo quase diário, sendo 56,9% e 54,1% respectivamente. Conclusão: Identificaram-se quatorze corantes nos rótulos dos produtos industrializados ultraprocessados. Destacaram-se os corantes artificiais caramelos III e IV, bordeuax S, amarelo crepúsculo e tartrazina, e naturais urucum e carmim.

Objetivo: describir los colorantes alimenticios presentes en los alimentos ultra-procesados consumidos por 273 estudiantes de una universidad pública de Río de Janeiro. Métodos: Fue caracterizado el perfil sociodemográfico y de salud, a partir de cuestionario semiestructurado. Consumo de alimentos ultra-procesados, fue obtenido a través del Cuestionario de Frecuencia Alimentar (gelatinas, galletas rellenas, caramelos y chicles, refrescos, preparados sólidos para refresco, jugos industrializados, condimentos preparados y pastas instantáneas) y los colorantes fueron identificados en las etiquetas de los productos. Resultados: El consumo de productos conteniendo colorantes como gelatinas, caramelos y chicles, refrescos y jugos industrializados fue superior al 80%. Caramelos, chicles y condimentos preparados tuvieron consumo casi diario, 56,9% y 54,1% respectivamente. Conclusión: Se identificaron catorce colorantes en las etiquetas de los productos industrializados ultra-procesados. Los colorantes artificiales caramelos III y IV, bordeuax S, amarillo crepúsculo y tartracina, y naturales urucum y carmim se destacaron.

A comparative evaluation of the staining capacity of microhybrid and nanohybrid resin-based composite to indian spices and food colorants: An In vitro study.

INTRODUCTION: Resin composite restorative materials can mimic the natural color and shade of the tooth. However, exogenous colorants from food and drinks can stain them due to adsorption. The influence of Indian food colorants and spices on resin composite restorations has not been evaluated extensively. AIM: This study aims to evaluate the staining capacity of microhybrid and nanohybrid resin-based composites, to saffron extract, tandoori powder, and turmeric powder. SUBJECTS AND METHODS: Forty samples of microhybrid (Kulzer Charisma) and nanohybrid (3M Filtek Z350) resin composites were prepared using an acrylic template of dimension 5 mm × 3 mm. They were randomly divided into four groups and immersed into solutions of saffron extract, tandoori powder, and turmeric powder. Distilled water was used as the control group. Color values (LFNx01, aFNx01, bFNx01) were measured by colorimeter using the CIE LFNx01aFNx01bFNx01 system before and after 72 h of immersion. Color differences ΔEFNx01ab were statistically analyzed. STATISTICAL ANALYSIS USED: Two-way ANOVA and post-hoc Tukey (honest significant difference) test were done using IBM SPSS Statistics for Windows, Version 19.0. Armonk, NY: IBM Corp. RESULTS AND DISCUSSION: : All the immersion media changed the color of the resin composites to varying degrees. However, turmeric solution showed the maximum mean color variation ΔEFNx01ab of 14.8 ± 2.57 in microhybrid resin composites and 16.8 ± 3.50 in nanohybrid resin composites. CONCLUSION: Microhybrid and nanohybrid resin composites tend to stain to Indian food colorants, especially to turmeric powder.

Occurrence of azo food dyes and their effects on cellular inflammatory responses.

OBJECTIVES: The aim of this study was to examine the occurrence of five azo food dyes-tartrazine, sunset yellow, carmoisine, allura red, and ponceau 4 R-in the food supply chain of Singapore and their effects on the in vitro synthesis of leukotriene B4 (LTB4) and F2-isoprostanes. METHODS: Trained personnel recorded the names of foods and beverages sold in a local supermarket that contained at least one of the five azo dyes. The occurrence of the azo dyes in the local food supply was computed. The synthesis of LTB4 and F2-isoprostanes from freshly isolated blood neutrophils were measured using gas chromatography-mass spectrometry. RESULTS: Of the 1681 processed food items, 194 (11.54%) contained at least one of the five azo dyes. Tartrazine was most prevalent in food and beverage products sold in Singapore, followed by allura red, sunset yellow, ponceau 4 R, and carmoisine. The five azo dyes augmented the in vitro synthesis of LTB4 and F2-isoprostanes from blood neutrophils. Tartrazine was significantly more potent in increasing LTB4 synthesis than the other dyes, which exhibited similar potencies. The five food dyes increased the formation of F2-isoprostanes from blood neutrophils at all tested concentrations. CONCLUSION: The high prevalence of azo dyes in the food supply of Singapore and their ability to elicit proinflammatory responses in vitro suggest a potential health risk to the local population.