In vitro and in vivo acute toxicity of an artificial butter flavoring.

Flavorings used in cookies, electronic cigarettes, popcorn, and breads contain approximately 30 chemical compounds, which makes it difficult to determine and correlate signs and symptoms of acute, subacute or chronic toxicity. The aim of this study was to characterize a butter flavoring chemically and subsequently examine the in vitro and in vivo toxicological profile using cellular techniques, invertebrates, and lab mammals. For the first time, the ethyl butanoate was found as the main compound of a butter flavoring (97.75%) and 24 h-toxicity assay employing Artemia salina larvae revealed a linear effect and LC50 value of 14.7 (13.7-15.7) mg/ml (R2 = 0.9448). Previous reports about higher oral doses of ethyl butanoate were not found. Observational screening with doses between 150-1000 mg/kg by gavage displayed increased amount of defecation, palpebral ptosis, and grip strength reduction, predominantly at higher doses. The flavoring also produced clinical signs of toxicity and diazepam-like behavioral changes in mice, including loss of motor coordination, muscle relaxation, increase of locomotor activity and intestinal motility, and induction of diarrhea, with deaths occurring after 48 h exposure. This substance fits into category 3 of the Globally Harmonized System. Data demonstrated that butter flavoring altered the emotional state in Swiss mice and disrupted intestinal motility, which may be a result of neurochemical changes or direct lesions in the central/peripheral nervous systems.

Effect of estragole over the RN4220 Staphylococcus aureus strain and its toxicity in Drosophila melanogaster.

Among the bacterial resistance mechanisms, efflux pumps are responsible for expelling xenobiotics, including bacterial cell antibiotics. Given this problem, studies are investigating new alternatives for inhibiting bacterial growth or enhancing the antibiotic activity of drugs already on the market. With this in mind, this study aimed to evaluate the antibacterial activity of Estragole against the RN4220 Staphylococcus aureus strain, which carries the MsrA efflux pump, as well as Estragole's toxicity in the Drosophila melanogaster arthropod model. The broth microdilution method was used to perform the Minimum Inhibitory Concentration (MIC) tests. Estragole was used at a Sub-Inhibitory Concentration (MIC/8) in association with erythromycin and ethidium bromide to assess its combined effect. As for Estragole's toxicity evaluation over D. melanogaster, the fumigation bioassay and negative geotaxis methods were used. The results were expressed as an average of sextuplicate replicates. A Two-way ANOVA followed by Bonferroni's post hoc test was used. The present study demonstrated that Estragole did not show a direct antibacterial activity over the RN4220 S. aureus strain, since it obtained a MIC ≥1024 µg/mL. The association of estragole with erythromycin demonstrated a potentiation of the antibiotic effect, reducing the MIC from 512 to 256 µg/mL. On the other hand, when estragole was associated with ethidium bromide (EtBr), an antagonism was observed, increasing the MIC of EtBr from 32 to 50.7968 µg/mL, demonstrating that estragole did not inhibited directly the MsrA efflux pump mechanism. We conclude that estragole has no relevant direct effect over bacterial growth, however, when associated with erythromycin, this reduced its MIC, potentiating the effect of the antibiotic.

Gamma-hexalactone flavoring causes DNA lesion and modulates cytokines secretion at non-cytotoxic concentrations.

BACKGROUND: The γ-hexalactone is a flavoring agent for alcoholic beverages, teas, breads, dairy products, coffees, buttery products among others. It presents low molecular weight and exhibits sweet fruity aroma with nuances of nuts. As far as we know, both literature and government regulations have gaps regarding the safe use of the γ-hexalactone. In this context, the main objective of this work was to evaluate the effects of γ-hexalactone through in silico and in vitro approaches. METHODS: The in silico analysis was performed through four free online platforms (admetSAR, Osiris Property Explorer®, pkCSM platform and PreADMET) and consisted of comparative structural analysis with substances present in databases. The computational prediction was performed in the sense of complement and guide the in vitro tests. Regarding in vitro investigations, screening of cytotoxicity (assessed by cell proliferation and viability parameters) in lymphocytes exposed to γ-hexalactone for 72 h were carried out previously to determine non-cytotoxic concentrations. Following this screening, concentrations of 5.15, 0.515, and 0.0515 µM were selected for the study of the respective potentials: genotoxic (assessed by DNA comet assay), chromosomal mutation (analysis of micronucleus frequency) and immunomodulatory (cytokine quantification using ELISA immunoassay). The results of in vitro assays were compared by one-way analysis of variance (ANOVA), followed by Bonferroni's post hoc test, conducted by statistic software. RESULTS: The platform PreADMET pointed out that γ-hexalactone is potentially mutagenic and carcinogenic. The comet assay data corroborate with these results demonstrating that γ-hexalactone at 5.15 µM caused lymphocytes DNA damage. In relation to cytokine secretion, the results indicate that lymphocytes were activated by γ-hexalactone at non-cytotoxic concentrations, involving an increase in the IL-1 levels in all tested concentrations, ranging from approximately 56 to 93%. The γ-hexalactone only at 5.15 µM induced increase in the levels of IL-6 (~ 60%), TNF-α (~ 68%) and IFN-γ (~ 29%), but decreased IL-10 (~ 46%) in comparison with the negative control (p < 0.05). No change was observed in total lymphocytes or in cell viability at the concentrations tested. CONCLUSIONS: In summary, the γ-hexalactone demonstrated immunomodulatory and genotoxic effects at non-cytotoxic concentrations in healthy lymphocytes.

Toxic, cytotoxic and genotoxic potential of synthetics food flavorings / Potencial tóxico, citotóxico y genotóxico de saborizantes sintéticos de alimentos

Food flavorings in general are few studied for the toxicological aspect. This condition justifies toxicity, cytotoxicity and genotoxicity assessments of the substances. In the present study, the toxicity of banana, cherry and hazelnut flavorings was evaluated in meristematic cells of roots of Allium cepa, in pure form (as marketed) and in the concentrations of 12.5; 25 and 50%, after 24 and 48 hours of exposure. Toxic potential of these food additives was also evaluated against Artemia salina nauplii at concentrations of 0.78; 1.56; 3.12; 6.25; 12.5; 25 and 50%, after 24 hours of exposure. The three additives, in all treatments and times of analysis considered, caused significant inhibition of cell division in A. cepa, however did not cause cellular alterations to the evaluated meristems. These food flavorings also caused significant mortality to micro crustaceans with LC50<100 μg/mL. From this, under the conditions of mentioned analyzes, cherry, banana and hazelnut flavorings induced significant toxicity and cytotoxicity to the bioassays used.

En general, los aspectos toxicológicos de los saborizantes de los alimentos son poco estudiados. Esta condición justifica las evaluaciones de toxicidad, citotoxicidad y genotoxicidad de estas sustancias. En el presente estudio, se evaluó la toxicidad de los aromas de plátano, cereza y avellana en células meristemáticas de raíces de Allium cepa, en forma pura (según comercializa) y en concentraciones de 12.5; 25 y 50%, después de 24 y 48 horas de exposición. El potencial tóxico de estos aditivos alimentarios también se evaluó frente a nauplios de Artemia salina a concentraciones de 0,78; 1,56; 3.12; 6.25; 12.5; 25 y 50%, después de 24 horas de exposición. Los tres aditivos, en todos los tratamientos y tiempos de análisis considerados, causaron inhibición significativa de la división celular en A. cepa, sin embargo, no causaron alteraciones celulares a los meristemos evaluados. Estos saborizantes alimentarios también causaron una mortalidad significativa a microcrustáceos con LC50 <100 μg/ mL. A partir de esto, bajo las condiciones de los análisis descriptos, los aromatizantes de cereza, plátano y avellana indujeron toxicidad significativa y citotoxicidad para los bioensayos utilizados.

Toxicity of synthetic flavorings, nature identical and artificial, to hematopoietic tissue cells of rodents / Toxicidade de aromatizantes alimentares sintéticos idênticos aos naturais e artificiais a células de tecido hematopoiético de roedores

The goal of this study was to analyze cytotoxicity, genotoxicity and mutagenicity to bone marrow cells of mice of nature identical synthetic flavorings, passion fruit and strawberry, and artificial synthetic flavorings, vanilla, chocolate, tutti-frutti and cookie, at doses 0.5; 1.0; 2.0; 5.0 and 10.0 mL/kg. The additives were given to the animals by gavage in a single daily application for seven days. Data were subjected to analysis of variance (ANOVA) followed by post Tukeys post hoc test, p < 0.05. Animals treated with 2.0; 5.0 and 10.0 mL/Kg of flavorings chocolate, strawberry and cookie, and 5.0 and 10.0 mL/Kg of flavorings vanilla and passion fruit died on the fifth and sixth day of the experiment, respectively. The doses 0.5 and 1.0 mL/Kg of the six additives significantly reduced erythropoiesis in the examined tissue. Also, treatments 0.5 and 1.0 mL/Kg of chocolate, and 1.0 mL/Kg of strawberry and biscuit induced the formation of micronuclei in the bone marrow erythrocytes, at a significant frequency. Therefore, under the study conditions, the six microingredients analyzed were cytotoxic and genotoxic, and additives strawberry, chocolate and cookie were also mutagenic in at least one of the evaluated doses.(AU)

Os aromatizantes são essenciais para a indústria na confecção de alimentos industrializados. Porém, pouco se sabe sobre o potencial tóxico desses microingredientes alimentares. Dessa forma, objetivou-se neste trabalho analisar, em células de medula óssea de camundongos, a citotoxicidade, genotoxicidade e mutagenicidade de aromatizantes alimentares sintéticos idênticos ao natural, de maracujá e morango, e artificiais, de baunilha, chocolate, tutti-frutti e biscoito, nas doses 0,5; 1,0; 2,0; 5,0 e 10,0 mL/Kg. Os aditivos foram administrados aos animais via gavagem em aplicação diária única durante sete dias. Os dados obtidos foram submetidos ao procedimento estatístico ANOVA com pós teste de Tukey, com p < 0.05. Os animais tratados com 2,0; 5,0 e 10,0 mL/Kg dos aromatizantes de chocolate, morango e biscoito, e 5,0 e 10,0 mL/Kg dos aromatizantes de baunilha e maracujá vieram a óbito no quinto e sexto dia de experimento, respectivamente. As doses 0,5 e 1,0 mL/Kg dos seis aditivos reduziram significativamente a eritropoiese do tecido analisado. Ainda, os tratamentos 0,5 e 1,0 mL/kg de chocolate, e 1,0 mL/Kg de morango e biscoito induziram a formação de micronúcleos aos eritrócitos de medula em frequência significante. Portanto, nas condições de estudo estabelecidas, os seis microingredientes analisados foram citotóxico e genotóxicos, e os aditivos de morango, chocolate e biscoito também foram mutagênicos em pelo menos uma das doses avaliadas.(AU)

Toxicity of synthetic flavorings, nature identical and artificial, to hematopoietic tissue cells of rodents / Toxicidade de aromatizantes alimentares sintéticos idênticos aos naturais e artificiais a células de tecido hematopoiético de roedores

Abstract The goal of this study was to analyze cytotoxicity, genotoxicity and mutagenicity to bone marrow cells of mice of nature identical synthetic flavorings, passion fruit and strawberry, and artificial synthetic flavorings, vanilla, chocolate, tutti-frutti and cookie, at doses 0.5; 1.0; 2.0; 5.0 and 10.0 mL/kg. The additives were given to the animals by gavage in a single daily application for seven days. Data were subjected to analysis of variance (ANOVA) followed by post Tukey's post hoc test, p <0.05. Animals treated with 2.0; 5.0 and 10.0 mL/Kg of flavorings chocolate, strawberry and cookie, and 5.0 and 10.0 mL/Kg of flavorings vanilla and passion fruit died on the fifth and sixth day of the experiment, respectively. The doses 0.5 and 1.0 mL/Kg of the six additives significantly reduced erythropoiesis in the examined tissue. Also, treatments 0.5 and 1.0 mL/Kg of chocolate, and 1.0 mL/Kg of strawberry and biscuit induced the formation of micronuclei in the bone marrow erythrocytes, at a significant frequency. Therefore, under the study conditions, the six microingredients analyzed were cytotoxic and genotoxic, and additives strawberry, chocolate and cookie were also mutagenic in at least one of the evaluated doses.

Resumo Os aromatizantes são essenciais para a indústria na confecção de alimentos industrializados. Porém, pouco se sabe sobre o potencial tóxico desses microingredientes alimentares. Dessa forma, objetivou-se neste trabalho analisar, em células de medula óssea de camundongos, a citotoxicidade, genotoxicidade e mutagenicidade de aromatizantes alimentares sintéticos idênticos ao natural, de maracujá e morango, e artificiais, de baunilha, chocolate, tutti-frutti e biscoito, nas doses 0,5; 1,0; 2,0; 5,0 e 10,0 mL/Kg. Os aditivos foram administrados aos animais via gavagem em aplicação diária única durante sete dias. Os dados obtidos foram submetidos ao procedimento estatístico ANOVA com pós teste de Tukey, com p < 0.05. Os animais tratados com 2,0; 5,0 e 10,0 mL/Kg dos aromatizantes de chocolate, morango e biscoito, e 5,0 e 10,0 mL/Kg dos aromatizantes de baunilha e maracujá vieram a óbito no quinto e sexto dia de experimento, respectivamente. As doses 0,5 e 1,0 mL/Kg dos seis aditivos reduziram significativamente a eritropoiese do tecido analisado. Ainda, os tratamentos 0,5 e 1,0 mL/kg de chocolate, e 1,0 mL/Kg de morango e biscoito induziram a formação de micronúcleos aos eritrócitos de medula em frequência significante. Portanto, nas condições de estudo estabelecidas, os seis microingredientes analisados foram citotóxico e genotóxicos, e os aditivos de morango, chocolate e biscoito também foram mutagênicos em pelo menos uma das doses avaliadas.

Toxicity of synthetic flavorings, nature identical and artificial, to hematopoietic tissue cells of rodents.

The goal of this study was to analyze cytotoxicity, genotoxicity and mutagenicity to bone marrow cells of mice of nature identical synthetic flavorings, passion fruit and strawberry, and artificial synthetic flavorings, vanilla, chocolate, tutti-frutti and cookie, at doses 0.5; 1.0; 2.0; 5.0 and 10.0 mL/kg. The additives were given to the animals by gavage in a single daily application for seven days. Data were subjected to analysis of variance (ANOVA) followed by post Tukey's post hoc test, p <0.05. Animals treated with 2.0; 5.0 and 10.0 mL/Kg of flavorings chocolate, strawberry and cookie, and 5.0 and 10.0 mL/Kg of flavorings vanilla and passion fruit died on the fifth and sixth day of the experiment, respectively. The doses 0.5 and 1.0 mL/Kg of the six additives significantly reduced erythropoiesis in the examined tissue. Also, treatments 0.5 and 1.0 mL/Kg of chocolate, and 1.0 mL/Kg of strawberry and biscuit induced the formation of micronuclei in the bone marrow erythrocytes, at a significant frequency. Therefore, under the study conditions, the six microingredients analyzed were cytotoxic and genotoxic, and additives strawberry, chocolate and cookie were also mutagenic in at least one of the evaluated doses.

Cigarette flavors, package shape, and cigarette brand perceptions: an experiment among young Brazilian women / Los sabores de cigarrillos, la forma del paquete y las percepciones de las marcas de cigarrillos: un experimento entre las jóvenes brasileñas / Sabores do cigarro, forma do pacote e percepções da marca do cigarro: um experimento entre mulheres jovens brasileiras

ABSTRACT Objective In 2012, a new Brazilian regulation prohibited the use of flavor additives in tobacco products. To better understand the potential impact of this regulation, this study examines how flavor descriptors on cigarette packaging influence brand perceptions among young Brazilian women. Methods An online cross-sectional experiment was conducted with Brazilian women aged 16-26 (N = 640: 182 smokers and 458 nonsmokers) who rated 10 cigarette packages from one of three conditions: 1) branded packs; 2) packs with the same size, shape, and verbal descriptions as in condition 1, but without brand imagery (i.e., "plain pack"); and 3) packs from condition 2 but without brand descriptors (i.e., "plain pack, no descriptors"). Mixed-effects linear regression models were utilized to determine what associations that pack features (i.e., experimental condition; flavor descriptor vs. not; slim pack vs. not) had with participant ratings of nine characteristics, including appeal, taste, smoothness, and attributes of people who smoke the brand. Results Flavored branded packs were rated as more appealing, better tasting, and smoother than flavored plain packs with descriptors. Compared to flavored plain packs with descriptors, the same packs without descriptors were rated less positively on eight of the nine characteristics. Compared to nonsusceptible nonsmokers, susceptible nonsmokers rated flavored packs more positively on eight of the nine characteristics. Slim packs were rated more positively than regular packs on eight of the nine characteristics. Conclusions Slim packs and brands highlighting tobacco flavors appear to increase positive perceptions of tobacco products. Banning tobacco flavorings and slim packs may reduce the appeal of smoking for young Brazilian women, as well as for other vulnerable populations.

RESUMEN Objetivo En 2012, una nueva regulación brasileña prohibió el uso de aditivos de sabor en los productos de tabaco. Para comprender mejor el impacto potencial de esta regulación, este estudio examina cómo los descriptores de sabor en los envases de cigarrillos influyen en las percepciones de marca entre las jóvenes brasileñas. Métodos Se realizó un estudio transversal, en línea con mujeres brasileñas de entre 16 y 26 años (N = 640: 182 fumadores y 458 no fumadores) que calificaron 10 paquetes de cigarrillos de acuerdo a una de las siguientes condiciones: 1) paquetes de marca; 2) paquetes con el mismo tamaño, forma y descripciones verbales que en la condición 1, pero sin imágenes de marca (es decir, "paquete simple"); y 3) paquetes de la condición 2 pero sin descriptores de marca (es decir, "paquete simple, sin descriptores"). Se utilizaron modelos de regresión lineal de efectos mixtos para determinar qué asociación existía entre las diferentes condiciones (es decir, condición experimental, descriptor de sabor vs. no, paquete delgado vs. no) con las calificaciones de los participantes de nueve características, incluyendo atractivo, sabor, suavidad y atributos de personas que fuman la marca. Resultados Los paquetes de marca con sabor se calificaron como más atractivos, de mejor sabor y más suaves que los paquetes simples con sabores y descriptores. En comparación con los paquetes simples con sabores con descriptores, los mismos paquetes sin descriptores fueron calificados de manera menos positiva en ocho de las nueve características. En comparación con los no fumadores no susceptibles, los no fumadores susceptibles calificaron los paquetes con sabor de forma más positiva en ocho de las nueve características. Los paquetes delgados fueron clasificados más positivamente que los paquetes regulares en ocho de las nueve características. Conclusiones Los paquetes delgados y las marcas que destacan los sabores del tabaco parecen aumentar las percepciones positivas de los productos de tabaco. Prohibir los sabores del tabaco y los paquetes delgados puede reducir el atractivo del tabaco para las jóvenes brasileñas, así como para otras poblaciones vulnerables.

RESUMO Objetivo Em 2012, uma nova regulamentação brasileira proibiu o uso de aditivos aromáticos nos produtos tabagísticos. Para entender melhor o impacto potencial desta regulação, este estudo examinou como a propaganda de aroma em embalagens de cigarros influencia a percepção das brasileiras jovens sobre a marca do produto. Método Estudo transversal em que 640 brasileiras (168 fumantes e 458 não fumantes) avaliaram online, uma entre três das seguintes características de 10 pacotes de cigarros: 1) embalagem com a imagem da marca; 2) pacotes com o mesmo tamanho, formato e descrição das características do produto 1, mas sem marca (havia apenas a descrição do produto) e, 3) embalagens nas condições 2, mas sem a descrição do produto (embalagem em branco). O modelo de regressão linear misto foi utilizado para determinar associações das características da embalagem do cigarro (Ex: situação experimental, descrição de aroma versus não, pacotes finos versus não) com as avaliações das participantes para nove características do produto, incluindo a apelação, aroma, suavidade e atributos de quem fuma certa marca. Resultados Pacotes aromatizados de marca foram classificados como mais atraentes, com melhor sabor e suavidade em relação aos que tinham simples descrição do produto. Os pacotes descritos como aromatizados comparados ao mesmo produto sem descrição obtiveram menores escores para oito dentre nove características avaliadas. Em comparação as não fumantes sem suscetibilidade ao tabagismo, as que eram suscetíveis classificaram produtos aromatizados mais positivamente em oito de suas nove características. As embalagens finas receberam julgamento mais positivo do que as com espessura regular para oito dentre nove itens analisados. Conclusões Pacotes finos e marcas que destacam os aromas do tabaco parecem aumentar a percepção positiva deste produto. Proibir a aromatização do tabaco e a oferta de pacotes finos de cigarros reduzem o apelo de fumar entre brasileiras jovens, bem como para outros grupos vulneráveis.

Antiproliferative and genotoxic effects of nature identical and artificial synthetic food additives of aroma and flavor / Efeito antiproliferativo e genotóxico de aditivos alimentares de aroma e sabor sintéticos, dos tipos idêntico ao natural e artificial

This study aimed to analyze the antiproliferative and genotoxic potential of synthetic food flavorings, nature identical passion fruit and artificial vanilla. This assessment used root meristem cells of Allium cepa L., in exposure times of 24 and 48 hours and using doses of 0.2; 0.4 and 0.6 mL. Roots were fixed in Carnoys solution, hydrolyzed in hydrochloric acid, stained with acetic orcein and analyzed with optical microscope at 400× magnification, 5,000 cells for each treatment. For data analysis, it was used Chi-square test at 5%. Doses of 0.2 mL at ET 48 h; 0.4 and 0.6 mL at ET 24 and 48 h of passion fruit flavor, and the three doses of the vanilla flavor at ET 24 and 48 h significantly reduced the cell division rate in the meristems of roots, proving to be cytotoxic. Doses of 0.2; 0.4 and 0.6 mL of the passion fruit additive, and the three doses of vanilla tested, in the two exposure times, induced mitotic spindle changes and micronuclei formation in the cells of the test organism used, proving to be genotoxic. Therefore, under the studied conditions, flavoring solutions of vanilla and passion fruit, marketed nationally and internationally, significantly altered the functioning of the cell cycle in root meristem cells of A. cepa.(AU)

Neste trabalho teve-se por objetivo analisar o potencial antiproliferativo e genotóxico de aromatizantes alimentares sintéticos, idêntico ao natural de Maracujá, e artificial de Baunilha. Esta avaliação foi realizada por meio das células meristemáticas de raízes de Allium cepa L., nos tempos de exposição de 24 e 48 horas e nas doses de 0,2; 0,4 e 0,6 ml. As raízes foram fixadas em solução de Carnoy, hidrolisadas em ácido clorídrico e coradas com orceína acética. Analisou-se, em microscópio óptico em aumento de 400×, 5.000 células por grupo tratamento, e utilizou-se o teste estatístico Qui-quadrado a 5% para análise dos dados. Verificou-se que as doses de 0,2 ml, no TE 48 h; 0,4 e 0,6 ml, nos TE 24 e 48 h, do aromatizante de Maracujá, e as três doses analisadas, nos TE 24 e 48 h, do aditivo de Baunilha reduziram significativamente o índice de divisão celular dos meristemas de raízes, mostrando-se citotóxicas. As doses 0,2; 0,4 e 0,6 ml do aditivo de Maracujá, e a de 0,6 ml do aromatizante de Baunilha, nos dois tempos de exposição considerados, induziram alterações de fuso mitótico e micronúcleos as células do organismo de prova utilizado, mostrando-se genotóxicas. Portanto, nas condições analisadas, as soluções aromatizantes de Baunilha e Maracujá, comercializadas nacional e internacionalmente, alteraram significativamente o funcionamento do ciclo celular das células meristemáticas de raízes de A. cepa.(AU)

Cytotoxicity of Cheese and Cheddar Cheese food flavorings on Allim cepa L root meristems / Citotoxicidade de aromatizantes alimentares de Queijo e Queijo Cheddar sob meristemas de raízes de Allim cepa L

Abstract Despite their great importance for the food industry, flavorings, in general, raise a number of questions regarding their cytotoxicity, mutagenicity and carcinogenicity, since, in the literature, there are few studies found evaluating the toxicity on the systemic and cellular level, of these chemical compounds. The root meristems of Allium cepa (onion) are widely used for the assessment of toxicity of chemical compounds of interest. Thus, this study aimed to evaluate, in A. cepa meristematic cells, individually and in combination at the cellular level, the toxicity of synthetic Cheese and Cheddar Cheese food flavorings, identical to the natural, at doses of 1.0 and 2.0 mL, at exposure times of 24 and 48 hours. In combination we used 0.5 mL of Cheese flavor associated with 0.5 mL of Cheddar flavor; and 1.0 mL of Cheese flavor associated with 1.0 mL of Cheddar flavor, at exposure times of 24 and 48 hours. For these evaluations, we used groups of five onion bulbs, which were first embedded in distilled water and then transferred to their respective doses. The root tips were collected and fixed in acetic acid (3:1) for 24 hours. The slides were prepared by crushing and were stained with 2% acetic orcein. Cells were analyzed throughout the cell cycle, totaling 5,000 for each control and exposure time. The mitotic indices calculated and cellular aberrations observed were subjected to statistical analysis using the chi-square test (p <0.05). No chromosomal abnormalities nor those of mitotic spindle were observed for the treatments performed. The results, both individually and in combination, showed that the flavorings under study significantly reduced the cell division rate of the test system cells used. Therefore, under the conditions studied, the two flavorings were cytotoxic.

Resumo Apesar da grande importância para a indústria alimentícia, os aromatizantes, em geral, suscitam uma série de dúvidas quanto a sua citotoxicidade, mutagenicidade e carcinogenicidade, visto que, na literatura, poucos são os trabalhos encontrados avaliando a toxicidade, em nível sistêmico e celular, destes compostos químicos. Os meristemas de raízes de Allium cepa (cebola) são muito utilizados para a avaliação da toxicidade de compostos químicos de interesse. Desta forma, este trabalho teve por objetivo avaliar em células meristemáticas de A. cepa, de forma individual, a toxicidade em nível celular de aromatizantes alimentares sintéticos, idênticos aos naturais, de sabores Queijo e Queijo Cheddar, nas doses de 1,0 e 2,0 mL, nos tempos de exposição de 24 e 48 horas; e de forma associada, onde se utilizou 0,5 mL do aromatizante sabor Queijo associado a 0,5 mL do aromatizante sabor Queijo Cheddar; e 1,0 mL do aromatizante sabor Queijo associado a 1,0 mL do aromatizante sabor Queijo Cheddar, nos tempos de exposição de 24 e 48 horas. Para estas avaliações utilizou-se grupos de cinco bulbos de cebolas, que primeiramente foram enraizados em água destilada, e em seguida transferidos para as suas respectivas doses. As radículas foram coletadas e fixadas em ácido acético (3:1) por 24 horas. As lâminas foram preparadas pela técnica de esmagamento e coradas com orceína acética a 2%. Analisaram-se células em todo ciclo celular, totalizando 5.000 para cada controle e tempo de exposição. Os índices mitóticos calculados e as aberrações celulares observadas foram submetidos à análise estatística do Qui-quadrado (p<0,05). Não foram observadas alterações cromossômicas e anomalias de fuso mitótico para nenhum dos tratamentos realizados. Os resultados obtidos, tanto individualmente como de forma associada, mostraram que os aromatizantes em estudos reduziram de forma significativa os índices de divisões celulares das células do sistema teste utilizado. Portanto, nas condições analisadas, os dois aromatizantes foram citotóxicos.

Cytotoxicity of Cheese and Cheddar Cheese food flavorings on Allim cepa L root meristems.

Despite their great importance for the food industry, flavorings, in general, raise a number of questions regarding their cytotoxicity, mutagenicity and carcinogenicity, since, in the literature, there are few studies found evaluating the toxicity on the systemic and cellular level, of these chemical compounds. The root meristems of Allium cepa (onion) are widely used for the assessment of toxicity of chemical compounds of interest. Thus, this study aimed to evaluate, in A. cepa meristematic cells, individually and in combination at the cellular level, the toxicity of synthetic Cheese and Cheddar Cheese food flavorings, identical to the natural, at doses of 1.0 and 2.0 mL, at exposure times of 24 and 48 hours. In combination we used 0.5 mL of Cheese flavor associated with 0.5 mL of Cheddar flavor; and 1.0 mL of Cheese flavor associated with 1.0 mL of Cheddar flavor, at exposure times of 24 and 48 hours. For these evaluations, we used groups of five onion bulbs, which were first embedded in distilled water and then transferred to their respective doses. The root tips were collected and fixed in acetic acid (3:1) for 24 hours. The slides were prepared by crushing and were stained with 2% acetic orcein. Cells were analyzed throughout the cell cycle, totaling 5,000 for each control and exposure time. The mitotic indices calculated and cellular aberrations observed were subjected to statistical analysis using the chi-square test (p <0.05). No chromosomal abnormalities nor those of mitotic spindle were observed for the treatments performed. The results, both individually and in combination, showed that the flavorings under study significantly reduced the cell division rate of the test system cells used. Therefore, under the conditions studied, the two flavorings were cytotoxic.

Toxicity in food flavorings at the cellular level associated with each other at different doses / Toxicidade em nível celular de aromatizantes alimentares associados entre si em diferentes doses

This study aimed to examine the cytotoxicity and genotoxicity of synthetic flavorings, nature identical, Chocolate, Strawberry and Condensed Milk. This evaluation was performed in root meristem cells of Allium cepa L., in exposure times of 24 and 48 hours and using doses of 0.2; 0.4 and 0.6 mL, in combination, in which one of the three doses of a flavoring was combined with a different dose of one of the two other flavor additives studied. Roots were fixed in Carnoys solution, hydrolyzed in hydrochloric acid, stained with acetic orcein and then analyzed, under light microscopy, 5,000 cells for each treatment. For data analysis, it was used Chi-square test at 5%. All the treatments with combinations between the flavorings Chocolate/Strawberry and Strawberry/Condensed Milk reduced, in both exposure times considered, cell division of A. cepa roots, proving to be cytotoxic. In turn, the treatments with the association of Chocolate/Condensed Milk did not change significantly the mitotic index of the cells analyzed. The Strawberry flavoring was the most cytotoxic among the additives tested. None of the evaluated associations was genotoxic under the study conditions.

Objetivou-se nesta pesquisa avaliar a citoxicidade e genotoxicidade de aromatizantes alimentares sintéticos de chocolate, morango e leite condensado. Esta avaliação ocorreu por meio das células meristemáticas de raízes de A. cepa L., nos tempos de exposição de 24 e 48h e nas doses de 0,2; 0,4 e 0,6 mL, em associação, em que para uma das três doses de um dos aromatizantes associou-se uma dose diferente de um dos outros dois aditivos de aroma em estudo. Em seguida, as raízes foram fixadas em solução de Carnoy, hidrolisadas em ácido clorídrico e coradas com orceína acética. Analisaram-se, em microscópio óptico, 5.000 células para cada grupo tratamento, e utilizou-se o teste estatístico Qui-quadrado a 5% para análise dos dados. A partir dos resultados, verificou-se que todos os tratamentos decorrentes das associações entre chocolate/morango e morango/leite condensado reduziram, nos dois tempos de exposição considerados, a divisão celular das raízes A. cepa, mostrando-se citotóxicos. Já os tratamentos provenientes da associação chocolate/leite condensado não alteraram de forma significativa os índices mitóticos das células do tecido em análise. Foi possível inferir que o aditivo de morango foi o mais citotóxico dos aditivos em estudo. Nenhuma das associações avaliadas foi genotóxica nestas condições de estudo.

Toxicity in food flavorings at the cellular level associated with each other at different doses / Toxicidade em nível celular de aromatizantes alimentares associados entre si em diferentes doses

This study aimed to examine the cytotoxicity and genotoxicity of synthetic flavorings, nature identical, Chocolate, Strawberry and Condensed Milk. This evaluation was performed in root meristem cells of Allium cepa L., in exposure times of 24 and 48 hours and using doses of 0.2; 0.4 and 0.6 mL, in combination, in which one of the three doses of a flavoring was combined with a different dose of one of the two other flavor additives studied. Roots were fixed in Carnoys solution, hydrolyzed in hydrochloric acid, stained with acetic orcein and then analyzed, under light microscopy, 5,000 cells for each treatment. For data analysis, it was used Chi-square test at 5%. All the treatments with combinations between the flavorings Chocolate/Strawberry and Strawberry/Condensed Milk reduced, in both exposure times considered, cell division of A. cepa roots, proving to be cytotoxic. In turn, the treatments with the association of Chocolate/Condensed Milk did not change significantly the mitotic index of the cells analyzed. The Strawberry flavoring was the most cytotoxic among the additives tested. None of the evaluated associations was genotoxic under the study conditions.(AU)

Objetivou-se nesta pesquisa avaliar a citoxicidade e genotoxicidade de aromatizantes alimentares sintéticos de chocolate, morango e leite condensado. Esta avaliação ocorreu por meio das células meristemáticas de raízes de A. cepa L., nos tempos de exposição de 24 e 48h e nas doses de 0,2; 0,4 e 0,6 mL, em associação, em que para uma das três doses de um dos aromatizantes associou-se uma dose diferente de um dos outros dois aditivos de aroma em estudo. Em seguida, as raízes foram fixadas em solução de Carnoy, hidrolisadas em ácido clorídrico e coradas com orceína acética. Analisaram-se, em microscópio óptico, 5.000 células para cada grupo tratamento, e utilizou-se o teste estatístico Qui-quadrado a 5% para análise dos dados. A partir dos resultados, verificou-se que todos os tratamentos decorrentes das associações entre chocolate/morango e morango/leite condensado reduziram, nos dois tempos de exposição considerados, a divisão celular das raízes A. cepa, mostrando-se citotóxicos. Já os tratamentos provenientes da associação chocolate/leite condensado não alteraram de forma significativa os índices mitóticos das células do tecido em análise. Foi possível inferir que o aditivo de morango foi o mais citotóxico dos aditivos em estudo. Nenhuma das associações avaliadas foi genotóxica nestas condições de estudo.(AU)

Toxicity at the cellular level of artificial synthetic flavorings / Toxicidade em nível celular de aromatizantes sintéticos artificiais

The goal of the present study was to evaluate the cytotoxicity and genotoxicity of artificial synthetic flavoring agents cookie and tutti-frutti. To this end, root meristem cells of Allium cepa L. were exposed to these substances in exposure times of 24 and 48 hour using individual doses of 0.3; 0.6 and 0.9 mL and doses combined as follows: 0.3 mL + 0.3 mL; 0.6 mL and 0.9 mL + 0.6 mL + 0.9 mL. After applying the treatments, root meristems were fixed, hydrolyzed, stained and analyzed a total of 5,000 cells using an optical microscope to evaluate each dose and combined treatment. All three doses of cookie flavoring and combined treatments significantly inhibited cell division of the tissue studied. Doses of tuttifrutti caused no change in cell division rate. In addition, doses of both flavorings and treatments combining these solutions induced cell aberrations in a significant number of cells to the A. cepa system. Therefore, under these analytical conditions, cookie flavoring and combined doses were cytotoxic and genotoxic, and tutti-frutti flavoring, although non-cytotoxic, demonstrated genotoxic action.(AU)

Objetivou-se neste trabalho avaliar a citotoxicidade e a genotoxicidade de aditivos aromatizantes sintéticos artificiais de biscoito e tutti-frutti. Essa avaliação se deu por meio das células meristemáticas de raízes de Allium cepa L., nos tempos de exposição de 24 e 48 horas, em que estas substâncias foram analisadas individualmente, nas doses de 0,3; 0,6 e 0,9 mL, e associadas entre si, da seguinte forma: 0,3 mL + 0,3 mL; 0,6 mL + 0,6 mL e 0,9 mL + 0,9 mL. Os meristemas de raízes foram fixados, hidrolisados e corados, e em seguida analisados em microscópio óptico, no qual se avaliou para cada dose e tratamento associado um total de 5.000 células. A partir dos resultados obtidos verificou-se que as três doses do aromatizante de biscoito e os tratamentos em associação inibiram significativamente a divisão celular dos tecidos em estudo. Já as doses de tutti-frutti não alteraram o índice de divisão celular das raízes em questão. Verificou-se também que as doses dos dois aromatizantes e os tratamentos em associação, induziram aberrações celulares em número significativo. Portanto, nestas condições de análise, o aditivo de biscoito e as doses associadas foram citotóxicos e genotóxicos, e o de tutti-frutti, apresentou apenas ação genotóxica.(AU)

Monosodium glutamate, a food additive, induces depressive-like and anxiogenic-like behaviors in young rats.

UNLABELLED: Monosodium glutamate (MSG) has been the target of research due to its toxicological effects. AIMS: We investigated the depressive- and anxiogenic-like behaviors in rats exposed to neonatal subcutaneous injection of MSG. The involvement of the serotonergic system, by measuring [(3)H] serotonin (5-HT) uptake in cerebral cortices, and the hypothalamic pituitary adrenal (HPA) axis, by determining serum adrenocorticotropic hormone (ACTH) and corticosterone levels, was also examined. MATERIALS AND METHODS: Male and female newborn Wistar rats were divided into control and MSG groups, which received, respectively, a daily subcutaneous injection of saline (0.9%) or MSG (4 g/kg/day) from the 1st to 5th postnatal day. The behavioral tests [spontaneous locomotor activity, contextual fear conditioning, and forced swimming test (FST)] were performed from the 60th to 64th postnatal day. MSG-treated animals showed alteration in the spontaneous locomotor activity, an increase in the number of fecal pellets and the number of animal's vocalizations and urine occurrence, and a decrease in the grooming time. KEY FINDINGS: The MSG exposure increased the immobility time in the FST and the freezing reaction in the contextual fear conditioning. Additionally, MSG treatment increased the [(3)H]5-HT uptake in the cerebral cortices of rats and induced a deregulation of HPA axis function (by increasing serum ACTH and corticosterone levels). SIGNIFICANCE: In conclusion MSG-treated rats are more susceptible to develop anxiogenic- and depressive-like behaviors, which could be related to a dysfunction in the serotonergic system.

Bronchiolitis associated with exposure to artificial butter flavoring in workers at a cookie factory in Brazil.

OBJECTIVE: To report the cases of four patients with bronchiolitis caused by exposure to artificial butter flavoring at a cookie factory in Brazil. METHODS: We described the clinical, tomographic, and spirometric findings in the four patients, as well as the lung biopsy findings in one of the patients. RESULTS: All four patients were young male nonsmokers and developed persistent airflow obstruction (reduced FEV1/FVC ratio and FEV1 at 25-44% of predicted) after 1-3 years of exposure to diacetyl, without the use of personal protective equipment, at a cookie factory. The HRCT findings were indicative of bronchiolitis. In one patient, the surgical lung biopsy revealed bronchiolitis obliterans accompanied by giant cells. CONCLUSIONS: Bronchiolitis resulting from exposure to artificial flavoring agents should be included in the differential diagnosis of airflow obstruction in workers in Brazil.

Bronquiolite associada à exposição a aroma artificial de manteiga em trabalhadores de uma fábrica de biscoitos no Brasil / Bronchiolitis associated with exposure to artificial butter flavoring in workers at a cookie factory in Brazil

OBJETIVO: Relatar quatro casos de bronquiolite decorrente de exposição a aroma artificial de manteiga em uma fábrica de biscoitos no Brasil. MÉTODOS: Descrevemos os achados clínicos, espirométricos e tomográficos nos quatro pacientes, assim como achados de biópsia pulmonar em um dos pacientes. RESULTADOS: Os quatro pacientes eram homens jovens, não fumantes, e desenvolveram obstrução persistente ao fluxo aéreo (relação VEF1/CVF reduzida e VEF1 de 25-44% do previsto) após 1-3 anos de exposição a diacetil, sem a utilização de equipamentos de proteção individual, em uma fábrica de biscoitos. A TCAR mostrou achados indicativos de bronquiolite. Em um paciente, a biópsia pulmonar cirúrgica mostrou bronquiolite obliterante associada a células gigantes. CONCLUSÕES: A bronquiolite decorrente de exposição a flavorizantes artificiais deve ser considerada em trabalhadores com obstrução ao fluxo aéreo no Brasil.

OBJECTIVE: To report the cases of four patients with bronchiolitis caused by exposure to artificial butter flavoring at a cookie factory in Brazil. METHODS: We described the clinical, tomographic, and spirometric findings in the four patients, as well as the lung biopsy findings in one of the patients. RESULTS: All four patients were young male nonsmokers and developed persistent airflow obstruction (reduced FEV1/FVC ratio and FEV1 at 25-44% of predicted) after 1-3 years of exposure to diacetyl, without the use of personal protective equipment, at a cookie factory. The HRCT findings were indicative of bronchiolitis. In one patient, the surgical lung biopsy revealed bronchiolitis obliterans accompanied by giant cells. CONCLUSIONS: Bronchiolitis resulting from exposure to artificial flavoring agents should be included in the differential diagnosis of airflow obstruction in workers in Brazil.