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

Abstract 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 Carnoy’s 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.

Resumo 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.

Meristem culture and subsequent micropropagation of Chilean strawberry (Fragaria chiloensis (L) Duch. )

BACKGROUND: Vegetative propagation of Fragaria sp. is traditionally carried out using stolons. This system of propagation, in addition to being slow, can spread plant diseases, particularly serious being viral. In vitro culture of meristems and the establishment of micropropagation protocols are important tools for solving these problems. In recent years, considerable effort has been made to develop in vitro propagation of the commercial strawberry in order to produce virus-free plants of high quality. These previous results can serve as the basis for developing in vitro-based propagation technologies in the less studied species Fragaria chiloensis. RESULTS: In this context, we studied the cultivation of meristems and establishment of a micropropagation protocol for F. chiloensis. The addition of polyvinylpyrrolidone (PVP) improved the meristem regeneration efficiency of F. chiloensis accessions. Similarly, the use of 6-benzylaminopurine (BAP) in the culture media increased the average rate of multiplication to 3-6 shoots per plant. In addition, the use of 6-benzylaminopurine (BAP), had low levels (near zero) of explant losses due to oxidation. However, plant height as well as number of leaves and roots were higher in media without growth regulators, with average values of 0.5 cm, 9 leaves and 4 roots per plant. CONCLUSIONS: For the first time in Chilean strawberry, meristem culture demonstrated to be an efficient tool for eliminating virus from infected plants, giving the possibility to produce disease free propagation material. Also, the addition of PVP into the basal MS medium improved the efficiency of plant recovery from isolated meristems. Farmers can now access to high quality plant material produced by biotech tools which will improve their technological practices.

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.

Evaluation of cytotoxicity and genotoxicity of Hancornia speciosa latex in Allium cepa root model / Avaliação da citotoxicidade e genotoxicidade do látex de Hancornia speciosa usando o modelo da raiz de Allium cepa

Abstract The latex obtained from Hancornia speciosa Gomes (Mangabeira tree) is widely used in traditional medicine to treat a variety of diseases, including diarrhea, ulcer, gastritis, tuberculosis, acne and warts. In this study, the cytotoxicity and genotoxicity effects of H. speciosa latex on the root meristem cells of Allium cepa were examined. Onion bulbs were exposed to different concentrations of latex and then submitted to microscopic analysis using Giemsa stain. Water was used as a negative control and sodium azide as a positive control. The results showed that, under the testing conditions, the mitotic index (MI) of the onion roots submitted to latex treatment did not differ significantly from the negative control, which suggests that the latex is not cytotoxic. Low incidence of chromosome aberrations in the cells treated with H. speciosa latex was also observed, indicating that the latex does not have genotoxic effect either. The MI and the chromosome aberration frequency responded to the latex concentration, requiring more studies to evaluate the dosage effect on genotoxicity. The results indicate that in tested concentrations H. speciosa latex is probably not harmful to human health and may be potentially used in medicine.

Resumo O látex obtido de Hancornia speciosa é amplamente utilizado na medicina popular para tratar uma variedade de doenças, tais como: diarreia, úlcera, gastrite, tuberculose, acne e verrugas. Nesse estudo, foram avaliados os efeitos citotóxicos e genotóxicos do látex de H. speciosa sobre as células meristemáticas das raízes de Allium cepa. Os bulbos das cebolas foram expostos a diferentes concentrações de látex e depois submetidos à analise microscópica usando o corante Giemsa. A água foi usada como controle negativo e a ázida sódica como controle positivo. Os resultados mostraram que o índice mitótico (IM) das raízes de cebola submetidas ao tratamento com látex, nas condições testadas, não diferiram significativamente do controle negativo, e sugerem que o látex não é citotóxico. Também foi observada uma baixa incidência de aberrações cromossômicas nas células tratadas com látex de H. speciosa, o que sugere que o látex também não possui efeito genotóxico. O IM e a frequência de aberrações cromossômicas foram dependentes da concentração de látex. Outros estudos devem ser realizados para avaliar o efeito da dose na genotoxidade. Os resultados indicam que o látex de mangabeira, nas concentrações testadas, provavelmente não é danoso para saúde humana e pode ter potencial para ser usado na medicina.

Genotoxicity and toxicity evaluations of ECF cellulose bleaching effluents using the Allium cepa L. Test / Genotoxicidade e toxicidade em Allium cepa L.: avaliação da qualidade do efluente no processo de branqueamento da celulose

Toxicity and genotoxicity tests were performed on root cells of Allium cepa in order to evaluate wastewater quality following an ECF cellulose bleaching process. The results revealed a toxic effect of the effluent, with inhibition of meristem growth and generally lower values of metaphase, anaphase and telophase indices at pH 10.5 than pH 7 for all effluent concentrations. The genotoxicity effect was different from the toxic effect given that the micronucleus and the chromosomal aberration tests in anaphase-telophase cells were low over all ranges of the studied effluent concentrations.

Os testes de toxicidade e genotoxicidade foram realizados em células da raiz de Allium cepa, a fim de avaliar a qualidade do efluente na sequência de um processo de branqueamento de celulose ECF. Os resultados revelaram um efeito tóxico do efluente, com inibição de crescimento do meristema e valores geralmente baixos de metáfase, anáfase e índices de telófase, a pH 10,5 e pH 7, para todas as concentrações do efluente. O efeito de genotoxicidade foi diferente do efeito tóxico, uma vez que o micronúcleo e os testes de aberrações cromossômicas em anáfase-telófase das células foram baixos em todas as gamas de concentrações do efluente estudado.

Cytogenotoxicity of Cymbopogon citratus (DC) Stapf (lemon grass) aqueous extracts in vegetal test systems

The lemon grass, Cymbopogon citratus (DC) Stapf, is an important species of Poaceae family commonly used in the folk medicine in many countries. The aim of this study was to investigate the cytotoxic and genotoxic effects of aqueous extracts from C. citratus leaves on Lactuca sativa (lettuce) root tip meristem cells by cytogenetic studies that have never been done before for lemon grass extracts. For this, lettuce seeds were treated for 72h with different concentrations of lemon grass aqueous extracts (5; 10; 20 and 30 mg/mL). The percentage of germination, root development and cellular behavior were analyzed, and the results showed that the highest concentration of aqueous extracts reduced the mitotic index, the seed germination and the root development of lettuce. The extracts have also induced chromosome aberrations and cellular death in the roots cells of L. sativa.

O capim-limão, Cymbopogon citratus (DC) Stapf, é uma importante espécie da família Poaceae com uma comum utilização na medicina popular em vários países. O objetivo deste estudo foi investigar os efeitos citotóxicos e genotóxicos do extrato aquoso das folhas de C. citratus em células meristemáticas de Lactuca sativa (alface) por meio de estudos citogenéticos, uma vez que estudos desta natureza não existem para extratos aquosos de capim-limão. Para isso, sementes de alface foram tratadas por 72h com diferentes concentrações de extratos aquosos feitos das folhas de capim-limão (5, 10, 20 e 30 mg/mL). O percentual de germinação, desenvolvimento radicular e o comportamento celular foram avaliados e os resultados mostraram que as concentrações mais elevadas dos extratos aquosos reduziram o índice mitótico, o percentual de germinação das sementes e desenvolvimento radicular da alface. Os extratos também induziram aberrações cromossômicas e morte celular nas células das raízes de L. sativa.

In vivo studies on the effect of Ocimum sanctum L. leaf extract in modifying the genotoxicity induced by chromium and mercury in Allium root meristems.

In vivo cytogenetic assay in Allium cepa root tip cells has been carried out to detect the modifying effect of Ocimum sanctum aqueous leaf extract against chromium (Cr) and mercury (Hg) induced genotoxicity. It was observed that the roots post-treated with the leaf extract showed highly significant (p < 0.001) recovery in mitotic index (MI) and chromosomal aberrations (CA) when compared to pre-treated (Cr/Hg) samples and the lower doses of the leaf extract were found to be more effective than higher doses. The present study reveals that the Ocimum sanctum leaf extract possesses the protective effect against Cr/Hg induced genetic damage.

A protocol for shoot multiplication from foliar meristem of Vanda spathulata (L.) Spreng.

Leaf explants collected from flowering plants of Vanda spathulata were cultured in Mitra medium with combinations of 6-benzyladenine (BA; 13.2-88.8 microM) and indole-3-acetic acid (IAA; 0.0 -85.6 microM). Combination of BA (66.6 microM) and IAA (28.5 microM) induced maximum shoots (17.33) from foliar meristems (leaf base). BA individually did not induce caulogenesis in leaf explants. For optimized multiplication, BA:IAA (2:1 microM) was essential at 22.2- 88.8 microM of BA. Re-cultured leaf explants produced lesser number of shoots compared to original explants and were nearly equal at combinations of 22.2-44.4 microM of BA and 5.7-28.5 microM of IAA. Rooting of shoots (> 95%) occurred in medium containing banana pulp (75 gl(-1)) and IAA (5.7 microM) within 3-9 weeks. Plantlets with 2-5 roots of 2-5 cm length established easily in community pots at 80-90% rates without hardening.

Assessment of the genotoxicity potential of the nematicide insecticide carbofuran in wheat cells

Carbofuran (Furadan), an insecticide-nematicide, is widely used for seed treatment in Brazil, against pests in crops such as wheat, rice and corn. It was assessed for its cytotoxic and or clastogenic potential in root meristems of Triticum aestivum (tested in vivo). Seed samples (1OO grams each) were individually treated with one, two, four and eight liters of the pure commercial product/lOO kg seed and germinated on three substrates: virgin soil, sterilized soil and growth chamber (on paper). For all treatments, no cytotoxic or clastogenic effects were observed in the meristem root cells.