POTENTIAL GENOTOXICITY AND HISTOPATHOLOGICAL ALTERATION EVALUATION OF HEPTEX®

Objective: The present study was performed in order to evaluate potential genotoxicity and the histopathological alteration of a traditional herbal prescription Heptex that used in the treatment of liver disease.Methods: The genotoxicity were evaluated using the in vivo chromosome aberration and micronucleus assays in bone marrow cells of male and female Sprague-Dawley rats. In addition, in vitro chromosome aberration assay in Chinese hamster ovary (CHO) cells and bacterial reverse mutations assay in Salmonella typhimurium strains and Escherichia coli (WP2-uvrA/) with and without metabolic activation system (S9 mix) were performed. Histopathological study was conducted in liver, ovary and testis tissue of Sprague-Dawley rats.Results: The genotoxicity assessment showed that Heptex did not significantly increase the number of chromosomal aberrations and frequencies of micro nucleated polychromatic erythrocytes in bone marrow cells of both male and female rats. In addition, there were no increases in the number of revertant colonies at any concentrations of Heptex used in the study. Heptex did not produce any structural aberration in CHO cells in the presence or absence of S9 mix. In addition, there were no histopathological changes induced by Heptex in rat liver, ovary and testis.Conclusion: Based on abovementioned findings, we can conclude that Heptex is generally non-toxic and does not exhibit genotoxicity or histopathological alteration.Â

Re‐evaluation of stannous chloride (E 512) as food additive

The Panel on Food Additives and Nutrient Sources added to Food (ANS) provides a scientific opinion re‐evaluating the safety of stannous chloride and stannous chloride dihydrate (E 512) as food additives. The Panel considered that adequate exposure and toxicity data were available. Stannous chloride is only permitted as food additives in one food category and no reply on the actual use level of stannous chloride (E 512) as a food additive and on its concentration in food was provided by any interested party. According to the Mintel's Global New Products Database (GNPD), stannous chloride was not labelled on any products in the EU nor in Norway. The regulatory maximum level exposure assessment scenario is based on the maximum permitted levels (MPLs) for stannous chloride (E 512), which is 25 mg Sn/kg. The mean exposure to stannous chloride (E 512) from its use as a food additive was below 1.3 μg Sn/kg body weight (bw) per day for all age groups. The 95th percentile of exposure to stannous chloride (E 512) ranged from 0.0 μg Sn/kg bw per day in all groups to 11.2 μg Sn/kg bw per day in adults. Absorption of stannous chloride from the gastrointestinal tract is low there is no concern with respect to carcinogenicity and genotoxicity. Gastrointestinal irritation was reported in humans after ingestion of a bolus dose of 40 mg Sn. The Panel concluded that stannous chloride (E 512) is of no safety concern in this current authorised use and use levels

Zaštitno djelovanje selenija protiv prekomjerne ekspresije apoptotskih gena povezanih s karcinomom u štakora izloženih o-krezolu

Cresols are monomethyl derivatives of phenol frequently used as solvents and intermediates in the production of disinfectants, fragrances, pesticides, dyes, and explosives, which is probably why they are widely distributed in the environment. General population may be exposed to cresols mainly through inhalation of contaminated air. In this study we evaluated the toxicological effects of o-cresol on differential gene expression profile of rat liver and prostate. Experiments were conducted on 80 male rats, 60 of which were exposed to o-cresol (1.5 g kg-1, 5 g kg-1, or 15 g kg-1) through feed for 8 weeks. Three groups of rats were supplemented with 0.1 mg kg-1 selenium (Se, in the form of, sodium selenite) in addition to o-cresol to evaluate its effectiveness against o-cresol toxicity. Control group received neither o-cresol nor Se, while one group received Se alone. Survival was similar between the exposed and control animals. Rats exposed to 15 g kg-1 of o-cresol showed a 16 % loss in body weight by the end of the study, which may have been related to o-cresol making feed unpalatable at this concentration. Liver and prostate tissue samples were collected at the end of the treatment. mRNA analysis revealed that apoptotic genes (CYP3A, COX-2, PPARγ, BAX, BCL2, AKT-1, and PKCα) related to cancer were up-regulated in liver and prostate tissues isolated from groups exposed to 5 g kg-1 and 15 g kg-1 o-cresol in comparison to control. Changes in gene expression profile were prevented when rats were supplemented with Se. The exact mechanisms underlying its protective effect remain to be clarified by future studies.Krezoli su monometilni derivati fenola koji se često rabe kao otapala te kao posrednici u proizvodnji dezinfekcijskih sredstava, mirisa, pesticida, boja i eksploziva. Otuda i njihova rasprostranjenost u okolišu. Opća je populacija izložena krezolima uglavnom putem zraka. U ovome se toksikološkom istraživanju ocijenilo djelovanje o-krezola, jednoga od tri krezolova izomera, na ekspresiju gena u tkivima jetre i prostate mužjaka štakora. Istraživanje je provedeno na 80 mužjaka, od kojih je 60 tijekom osam tjedana bilo izloženo o-krezolu (1,5 g kg-1, 5 g kg-1, odnosno 15 g kg-1) preko krmiva. Tri skupine štakora primale su uz o-krezol nadomjestak selenija u dozi od 0.1 mg kg-1 (Se, u obliku natrijeva selenita) radi ocjene njegove djelotvornosti protiv toksičnosti o-krezola. Kontrolna skupina nije primala ni o-krezol ni Se, dok je jedna skupina primala samo Se. Preživljenje je bilo podjednako u svih skupina životinja. Štakori izloženi najvišoj dozi o-krezola (15 g kg-1) imali su 16 % manju tjelesnu masu od kontrolne skupine na kraju ispitivanja, što može biti povezano s lošim okusom krmiva zbog primjese visoke doze o-krezola. S istekom osmotjednoga izlaganja o-krezolu životinje su eutanazirane te su prikupljeni uzorci tkiva jetre i prostate. Analiza m-RNA pokazala je značajno povišenu ekspresiju apoptotskih gena CYP3A, COX-2, PPARγ, BAX, BCL2, AKT-1 i PKCα, koji su povezani s nastankom karcinoma u skupinama štakora izloženim o-krezolu (5 g kg-1 i 15 g kg-1 u odnosu na kontrolu. Ova je prekomjerna ekspresija poništena u štakora koji su primali selenij. Još nisu jasni mehanizmi iza ovoga zaštitnog djelovanja, na što će odgovoriti buduća istraživanja

Flaxseed oil as a protective agent against bisphenol-A deleterious effects in male mice

Abstract Background Flaxseed oil is one of the most vital oils that contain a high content of omega-3 polyunsaturated fatty acids (PUFAs) recognized as a high-quality nutrition for health benefits. In addition, a source of vitamin E and β-carotene has a positive health effect in a variety of pathologies. The goal of the present work was to investigate the protecting character of flaxseed oil against bisphenol-A deleterious effects in male mice. Animals were gavaged with BPA for 28 successive days. They were orally administered flaxseed oil either before, with, or after treatment of BPA. DNA damage was evaluated in the liver, testes, and bone marrow cells using comet and micronucleus assays. Liver and testes histopathological examination as well as sperm physical characters were also investigated. Results The results showed that BPA induced a significant raise in DNA damage that obviously appear in the increase of tail length, tail DNA percentage, and tail moment in the liver and testes. In addition, there was an increase in the frequency of micronuclei in bone marrow cells. Liver and testes histopathological alterations and sperm count and motility significant comedown were seen in male mice exposed to BPA as well. Conversely, flaxseed oil oral administration through the three regimens of treatment effectively attenuated the abovementioned effects. Moreover, administration of flaxseed oil before BPA treatment was the best protocol in the attenuation of BPA toxic effects. Conclusions Flaxseed oil successfully attenuated the BPA genotoxicity, sperm defects, and histological alterations in male mice that may be referred to its antioxidant property

Genotoxic evaluation for the estrogenic mycotoxin zearalenone

Genotoxic effects of the mycotoxin Zearalenone (ZEN) were evaluated on albino mice. The investigation was assessed using 4 criteria: chromosome aberrations in bone marrow and spermatocytes of adult male mice; chromosome analysis and teratological effects of mice embryos. Zearalenone was administrated to both adult males and pregnant females with 2 doses level (5 $\mu$g$\cdot$kg$^{-1}$ and 10 $\mu$g$\cdot$kg$^{-1}$ ZEN). Zearalenone was found to reduce the mitotic activity in treated males and the embryos proving that it is a cytotoxic substance. In treated males and females, it induced some chromosome abnormalities with no significant increase over the control at the doses investigated, except for some few figures. Similar results were observed for the teratological study. The results in general could consider zearalenone as a toxic mycotoxin for both adult animals and embryos. It is highly recommended that a great attention should be paid towards the toxicity of zearalenone to mono-gastric animals and human, especially it contaminate corn that is widely used in human and animal feeding

Genotoxical, teratological and biochemical effects of anthelmintic drug oxfendazole Maximum Residue Limit (MRL) in male and female mice

Oxfendazole, methyl-5 (6)-phenylsulfinyl-2-benzimidazole carbamate, is a member of the benzimidazole family of anthelmintics. Anthelmintic benzimidazoles are widely used in meat producing animals (cattle, sheep and pigs) for control of endoparasites. The extensive use of veterinary drugs in food-producing animals can cause the presence of small quantities of the drug residues in food. Maximum residue limit or “MRL” means the maximum concentration of residue resulting from the use of a veterinary medicinal product which may be legally permitted recognized as acceptable in food. The FAO/WHO Expert Committee on Food Additives (1999) evaluations of toxicological and residue data, reported that oxfendazole (MRL) has toxicological hazards on human health. The toxicity of oxfendazole (MRL) was tested in male and female mice and their fetuses. Chromosomal aberrations, teratological examination and biochemical analysis were the parameters used in this study. The results show that oxfendazole MRL induced a mutagenic effect in all tested cell types. Also, oxfendazole exhibit embryotoxicity including teratogenicity. The biochemical results show that oxfendazole induced a disturbance in the different biochemical contents of all tested tissues. So, we must increase the attention paid to the potential risk of oxfendazole residues in human beings and should stress the need for careful control to ensure adherence to the prescribed withdrawal time of this drug

Improved effect of pumpkin seed oil against the bisphenol-A adverse effects in male mice

The present study was conducted to evaluate the ameliorative role of pumpkin seed oil (PSO) against potential adverse effects of bisphenol-A (BPA) in male mice. BPA was administered to the mice orally at a dose of 50 mg/kg body weight once a day for 28 successive days. While, PSO was administered to the mice orally at 1 mL/kg b w either before, with or after treatment of BPA, once a day for 28 successive days. The studied parameters were DNA damage evaluation using comet assay in liver and testes cells and micronucleus test in bone marrow; and histopathological examination of liver and testes tissues. Results revealed that BPA induced DNA damage in tested cells and marked histopathological alterations in liver and testes. In contrast, PSO treatments alleviated DNA damage and improved the histopathological alterations in liver and testes tissues. Furthermore, administration of mice with the PSO before BPA treatment was the best regimen in the alleviation of the adverse effects of BPA, followed by administration of PSO after then with treatment of BPA. It can be concluded that PSO may has a protective role against BPA genotoxicity and histopathological alterations in male mice. Keywords: Pumpkin seed oil, Bisphenol-A, Genotoxicity, Micronucleus test, Comet assay, Histopatholog

Genotoxicity of Chlorpyrifos and the Antimutagenic Role of Lettuce Leaves in Male Mice

Chlorpyrifos [O O-diethyl-O-(3 5 6-trichloro-2-pyridyl)-phosphorothioate] is one of the most widely used organophosphate insecticides. Previous studies proved that chlorpyrifos, at different doses, induced genotoxicity. In Egyptian foods, the residual levels of pesticides are often higher than those found in developed country ones. So the aim of this research was to evaluate the genotoxicity of the insecticide chlorpyrifos at doses equal to its maximum residue limit (MRL) in the leafy vegetables, its double and quadruple (0.5, 1 and 2 mg/kg body weight) in somatic and germ cells of male mice. In addition to that, evaluating the role of lettuce leaves as antigenotoxic in reducing the genotoxic effects of chlorpyrifos tested doses when concurrently administrated to these animals. The study was conducted on adult male laboratory mice at three levels: bone marrow cells as a model for mitotic chromosome aberrations, spermatocytes as a model for meiotic chromosomes and sperm count and morphology. The results of the present study indicate that the treatment of male mice with chlorpyrifos by oral gavages for three months induced significant increase in the frequencies of total chromosomal aberrations in both somatic and germ cells in relation to control groups. Results of the sperm analysis showed that chlorpyrifos induced significant decrease in the sperm count when compared to negative control. Furthermore, it induced significant increase in head and tail sperm abnormalities, among which coiled tail was considered the most obvious sperm abnormality induced by chlorpyrifos. At the same time, the present study indicated that lettuce leaves feed concurrently with three doses of chlorpyrifos could not protect cells from damage.Chlorpyrifos [O O-diethyl-O-(3 5 6-trichloro-2-pyridyl)-phosphorothioate] is one of the most widely used organophosphate insecticides. Previous studies proved that chlorpyrifos, at different doses, induced genotoxicity. In Egyptian foods, the residual levels of pesticides are often higher than those found in developed country ones. So the aim of this research was to evaluate the genotoxicity of the insecticide chlorpyrifos at doses equal to its maximum residue limit (MRL) in the leafy vegetables, its double and quadruple (0.5, 1 and 2 mg/kg body weight) in somatic and germ cells of male mice. In addition to that, evaluating the role of lettuce leaves as antigenotoxic in reducing the genotoxic effects of chlorpyrifos tested doses when concurrently administrated to these animals. The study was conducted on adult male laboratory mice at three levels: bone marrow cells as a model for mitotic chromosome aberrations, spermatocytes as a model for meiotic chromosomes and sperm count and morphology. The results of the present study indicate that the treatment of male mice with chlorpyrifos by oral gavages for three months induced significant increase in the frequencies of total chromosomal aberrations in both somatic and germ cells in relation to control groups. Results of the sperm analysis showed that chlorpyrifos induced significant decrease in the sperm count when compared to negative control. Furthermore, it induced significant increase in head and tail sperm abnormalities, among which coiled tail was considered the most obvious sperm abnormality induced by chlorpyrifos. At the same time, the present study indicated that lettuce leaves feed concurrently with three doses of chlorpyrifos could not protect cells from damage.Genotoxidade de Chlorpyrifos e papel antimutagênico das folhas de alfaces em ratosResumoChlorpyrifos [O O-diethyl-O-(3 5 6-trichloro-2-pyridyl)-phosphorothioate] é um dos inseticidades a basede organofosfato mais utilizados. Estudos anteriores provaram que chlorpyrifos, em doses diferentes,induziu a genotoxidade. Em alimentos egípcios, os níveis residuais de pesticidas são normalmentemais elevados que aqueles encontrados em países desenvolvidos. Então, o objetivo dessa pesquisafoi o de avaliar a genotoxidade do inseticida chlorpyrifos em doses iguais ao limite residual máximo(MRL) em vegetais com folhas, seu dobro e quádruplo (0.5, 1 and 2 mg/kg peso corpóreo) emcélulas somáticas e germinativas de ratos. Além disso, avaliando o papel das folhas de alfacecomo antígenos tóxicos na redução dos efeitos de genotoxidade em doses de chlorpyrifos testadasquando administradas a esses animais. O estudo foi conduzido em ratos adultos de laboratório, emtrês níveis: células ósseas como modelo para aberrações miótico-cromossômicas; espermatócitoscomo modelo meiótico-cromossômico, contagem e morfologia de espermatozóides. Os resultadosdo presente estudo indicaram que o tratamento de ratos com chlorpyrifos, administrado emdoses orais por três meses induziram um significativo aumento da freqüência de aberraçõescromossômicas totais tanto em células somáticas quanto em células germinativas se relacionadosaos grupos de controle. Os resultados da análise de espermatozóide demonstraram que chlorpyrifosinduziu uma diminuição significativa na contagem de espermatozóides quando comparado aogrupo de controle negativo. Além disso, também houve a indução de um aumento significativoem anomalias na cabeça e na cauda dos espermatozóides, dentre as quais a cauda enroladafoi considerada a anomalia mais óbvia entre os espermatozóides induzidos por chlorpyrifos. Aomesmo tempo, o presente estudo indicou que as folhas de alface alimentadas regularmente comtrês doses de chlorpyrifos não conseguiram proteger suas células de danos.