In vitro approaches to antioxidant screening for the development of a sunscreen formulation

Abstract The incorporation of antioxidants into sunscreens may provide additional skin photoprotection against the harmful photobiological effects of ultraviolet radiation. The present study evaluated the applicability of a screening approach to the assessment of the antioxidant and photoprotective properties of vitamin C, vitamin E, and coenzyme Q10 and then determined the performance of the most effective antioxidant in a sunscreen formulation. Antioxidant activity was assessed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, 2,2`-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay, and oxygen radical absorbance capacity (ORAC) assay, and the photoprotective potential was investigated by the yeast photoprotection assay. The antioxidant with the best effect was incorporated into sunscreen formulations which were evaluated for 120 days regarding their in vitro photoprotective parameters. Vitamin C showed high antioxidant capacity as well as a photoprotective potential against simulated solar irradiation applied for times longer than 1 h. Although the Sun Protection Factor, UVA/UVB ratio and critical wavelength did not differed significantly (p<0.05) between the formulation blank and the formulations containing 0.5% or 1% vitamin C, formulations with vitamin C kept their photostability for 6 months. Consequently, the proposed screening approach seems to be promising for the development of an antisolar photostable formulation containing vitamin C as an antioxidant.

Heteroisotherapics effect on the treatment against smoking: a cognitive-behavioral approach

The most common way of consuming nicotine is in tobacco cigarettes. Nicotine causes intense addiction. The National Cancer Institute coordinates and executes the Tobacco Control Program in Brazil, through actions that encourage the adoption of healthier lifestyles. In this context, homeopathy has used Heteroisotherapic medicines formulated according to the homeopathic pharmaceutical technology with scientific evidence of efficacy in the detoxification of substances and metals, and in the desensitization of foods or medicines. Aims Promote the importance of the cognitive-behavioral approach in combination with the homeopathic treatment against smoking. Methodology In the initial phase of the randomized double-blind clinical study (CEP / HUCFF / UFRJ 65622916.2.0000.5257), the effectiveness of the 6CH heteroisotherapeutic drug was assessed. Volunteers were recruited andin-person welcoming meetings, using the cognitive-behavioral approach, were carried out to inform them about the risks of smoking and the benefits of quitting. In addition, they were supported and guided during the smoking cessation process so that they could deal with the withdrawal syndrome, the psychological dependence and the constraints associated with smoking. Results and discussion84 participants were selected according to the inclusion criteria, and divided by randomization into two groups:the Test Group (heteroisotherapeutic medication) and the Control Group (homeopathic medication Nux vomica6CH).Both groups will be followed for 12 months. The combination of the following approaches has led to a significant increase in the cessation rate: I.Prepare the smoker for solving his own issues; II. Stimulate skills to resist temptations to smoke; III. Prepare to prevent relapse; IV. Prepare to deal with stress. Studies show that, regardless the duration of these approaches, there is an increase in the abstinence rate. Moreover, the longer the total approach time (frequency multiplied by the time spent on each contact), the higher the abstinence rate. On the other hand, from a total approach time of 90 minutes on, there is no further increase in the abstinence rate. ConclusionThe partial results obtained so far demonstrate that the cognitive-behavioral approach played a decisive role in the groups performance, favoring the treatment adherence as well as the group cohesion around the Project's objective, contributing to the effectiveness of the medicine, a decreased anxiety, improved sleep, cessation or decrease in the number of cigarettes smoked per day and the abstinence rate.

Avaliação do potencial genotóxico e mutagênico de um nosódio vivo obtido com o vírus Influenza A / Evaluation of the genotoxic and mutagenic potentials of a living nosode compounded with Influenza A virus

Introdução: O vírus da gripe tem sido responsável por doenças respiratórias contagiosas com altas taxas de mortalidade [1]. Algumas drogas tem sido usadas para tratar a gripe humana. No entanto, esses medicamentos causam muitos efeitos colaterais e induzem o aparecimento de cepas virais resistentes [2]. O impacto causado pelo vírus influenza tem motivado o desenvolvimento de novas abordagens para a prevenção e controle da influenza [3]. Portanto, um novo medicamento homeopático foi desenvolvido utilizando, como ponto de partida, o vírus influenza infecciosa [4]. Este pertence a um grupo chamado nosódios vivos [5]. No entanto, seus potenciais mutagênicos e genotóxicos, especialmente quando usado em diluições baixas, ainda não foram avaliados e é importante porque este bioterápico é preparado a partir de microorganismos vivos. Diferentes métodos podem ser usados para detectar efeitos mutagênicos e genotóxicos. Objetivos: Este estudo visa avaliar o potencial genotóxico e mutagênico do nosódio vivo do vírus influenza A, em diferentes potências homeopáticas.Metodologia: 1 ml de suspensão viral purificada foi diluída em 9 ml de água destilada estéril. Esta amostra foi submetida a 100 sucussões mecânicas (aproximadamente 3 Hz), usando o aparato Autic ® brasileira, originando a primeira diluição, chamada decimal (1x). 1 ml desta solução foi diluída em 9 ml de solvente e foi submetido a 100 sucussões, gerando o bioterápico 2x. Este procedimento foi repetido sucessivamente, de acordo com a Farmacopéia Homeopática Brasileira, para obter o bioterápico 30x [6]. Pela mesma técnica, a água foi preparada até a potência 30x, para ser utilizada como controle. Todas as amostras foram preparadas sob condições estéreis e assépticas, utilizando-se fluxo laminar, classe II, e foram armazenados em geladeira (8ºC). As amostras 1x, 6x, 12x, 18x 24x, 30x e 30x e água (controle do veículo) foram analisadas por: Induteste, avalia a capacidade dos agentes físicos ou químicos de promover a indução lisogênico como um reflexo dos danos nas moléculas de DNA em bactérias lisogênicas, e o teste de Ames, que utiliza linhagens indicadoras de Salmonella typhimurium, sensíveis a substâncias que podem induzir diferentes tipos de mutação. Resultados: Os resultados obtidos no Induteste não mostraram diminuição da fração de sobrevivência das bactérias utilizadas, e nenhum aumento na formação de indução lisogênica, em quaisquer potências testadas. O mesmo perfil foi obtido após o teste de Ames, com resultados semelhantes ao controle negativo. Conclusão: Conclui-se que nosódio vivo obtido com vírus Influenza A não é capaz de induzir danos no DNA de células procarióticas. Este resultado nos permite concluir que pacientes que usam este medicamento não tem efeitos colaterais relacionados com a mutagênese e genotoxicidade.(AU)

Background: The influenza virus has been responsible for contagious respiratory diseases with high mortality rates [1]. Some drugs have been used to treat human influenza. However, these drugs cause many common side effects and induce the appearance of resistant viral strains [2]. The impact caused by the influenza virus has motivated the development of new approaches for the prevention and control of influenza [3]. Therefore, a new homeopathic medicine was developed using, as a starting point, the infectious influenza virus [4]. This belongs to a group called living nosodes [5]. However, its mutagenic and genotoxic potentials, especially when used in low dilutions, has not yet been evaluated and it is important because this biotherapic is prepared from living microorganisms. Different methods can be used to detect mutagenic and genotoxicic effects. Aims: This study aims to evaluate the genotoxic and mutagenic potentials of influenza A living nosode at different homeopathic potencies. Methodology: 1 ml of purified viral suspension was diluted in 9 ml of sterile distilled water. This sample was submitted to 100 mechanical succussions (approximately 3 Hz), using Autic® Brazilian machine, originating the first dilution, named decimal (1x). 1 ml of this solution was diluted in 9 ml of solvent and was submitted to 100 sucussions, generating biotherapic 2x. This procedure was successively repeated, according to Brazilian Homeopathic Pharmacopoeia, to obtain the biotherapic 30x [6]. By the same technique, water vehicle was prepared until 30x potency to be used as control. All samples were prepared in sterile and under aseptic conditions, using laminar flow cabinet, class II, and were stored in the refrigerator (8ºC). The samples 1x, 6x, 12x, 18x, 24x and 30x and water 30x (vehicle control) were analysed by: the Inductest, which assesses the ability of physical or chemical agents to promote lysogenic induction as a reflection of damage in DNA molecules in lysogenic bacteria, and the Ames test, which uses indicator strains of Salmonella typhimurium, sensitive to substances that can induce different types of mutation. Results: The Inductest showed no decrease in the survival fraction of the bacteria used, and no increase in the formation of lysogenic induction, in any tested potency. The same profile was obtained after the Ames test, with similar results to negative control. Conclusion: We can conclude that this living nosode compounded with Influenza A virus is not able to induce DNA damage in prokaryotic cells. This result permits us to conclude that patients who use this medicine have no side effects related to mutagenesis and genotoxicity.(AU)

Avaliação do potencial genotóxico e mutagênico de soluções homeopáticas de Candida albicans / Evaluation of the genotoxic and mutagenic potentials of homeopathic Candida albicans solutions

ntrodução: Candida spp é encontrada naturalmente na microbiota natural da pele humana, trato gastrointestinal e genitourinário e, em geral, até 75% da população não apresenta qualquer sintoma [1]. No entanto, a candidíase oral é muito comum entre os pacientes HIV e pacientes submetidos à quimioterapia. O tratamento da candidíase oral é necessário, uma vez que a doença provoca desconforto e disfagia, resultando em má nutrição, recuperação lenta e internação prolongada [2,3]. Os resultados preliminares obtidos pelo nosso grupo com um novo bioterápico preparado a partir de Candida albicans (Candida 30x) mostrou um grande potencial para reduzir a taxa de adesão da candida quando células epiteliais foram pré-tratadas. Este estudo está sendo desenvolvido visando avaliação do potencial mutagênico e genotóxico de várias soluções homeopáticas.Objetivos: O objetivo deste estudo foi avaliar o potencial genotóxico e mutagênico de diferentes potências homeopáticas do medicamento de C. albicans. Metodologia: Uma parte de C. albicans obtida do sangue de pacientes brasileiros [4] foi diluída em 9 partes de água estéril. Esta amostra foi submetida a 100 sucussões mecânicas (aproximadamente 3 Hz), usando Autic ®, originando a primeira diluição (1x). Em seguida, 1 ml desta solução foi diluída em 9 ml de solvente, submetida a 100 sucussões, obtendo a potência 2x. Este procedimento foi repetido sucessivamente para obter a potência de 30x, de acordo com a Farmacopéia Homeopática Brasileira [5]. Pela mesma técnica, o veículo foi preparado até 30x para ser usado como controle. Todas as amostras foram preparadas em condições estéreis e assépticas, utilizando-se gabine de fluxo laminar, classe II, e foram armazenados em geladeira (8ºC). As amostras 1x, 6x, 12x, 18x 24x, 30x de C. albicans e 30x de água (controle do veículo) foram analisadas por: Induteste, que avalia a capacidade de agentes físicos ou químicos em promover a indução lisogênica em reflexo a danos em moléculas de DNA de bactérias lisogênicas. E o Teste de Ames, que utiliza linhagens indicadoras de Salmonella typhimurium, sensíveis a substâncias que podem induzir diferentes tipos de mutação.Int J High Dilution Res 2011; 10(36):177-179Proceedings of the XXV GIRI Symposium and VIII CBFH; 2011 Sep 04-07; Foz do Iguaçu (Brazil)179Resultados: De acordo com o Induteste, não houve diminuição da fração de sobrevivência de bactérias e aumento na formação de centros infecciosos, independentemente da potência homeopática testada. O mesmo perfil foi obtido após o Teste de Ames, com resultados semelhantes ao controle negativo. Conclusão: Podemos concluir que estas amostras não são capazes de induzir danos ao DNA bacteriano das cepas utilizadas. Assim, a utilização deste medicamento não apresenta quaisquer efeitos secundários relacionados com a mutagênese e genotoxicidade.(AU)

Background: Candida spp is naturally found in humans? flora of skin, gastrointestinal and genitourinary tracts and, in general, up to 75% of the population does not have any symptom [1]. However, oral candidiasis is very common among HIV patients and patients undergoing chemotherapy. The treatment of oral candidiasis is necessary once the disease causes discomfort and dysphagia, resulting in poor nutrition, slow recovery, and prolonged hospital stay [2,3]. Preliminary results obtained by our group with a new biotherapic prepared from Candida albicans (Candida 30x) showed a great potential to reduce the candida yeast adhesion rate when the epithelial cells were pre-treated. This study is currently being developed with the evaluation of mutagenic and genotoxic potentials of several homeopathic solutions. Aims: The goal of this study was to assess the genotoxic and mutagenic potentials of different homeopathic potencies of C. albicans. Methodology: One part of C. albicans yeast obtained from Brazilian patient?s blood [4] was diluted in 9 parts of sterile water. This sample was submitted to 100 mechanical succussions (approximately 3 Hz), using Autic® Brazilian machine, originating the first dilution (1x). Then, 1 ml of this solution was diluted in 9 ml of solvent, submitted to 100 succussions, obtaining 2x potency. This procedure was successively repeated to obtain 30x potency, according to Brazilian Homeopathic Pharmacopoeia [5]. By the same technique, water vehicle was prepared until 30x to be used as control. All samples were prepared in sterile and aseptic conditions, using laminar flow cabinet, class II and were stored in the refrigerator (8ºC). The samples 1x, 6x, 12x, 18x, 24x and 30x of C. albicans and water 30x (vehicle control) were analysed by: the Inductest, which assesses the ability of physical or chemical agents to promote lysogenic induction as a reflection of damage in DNA molecules in lysogenic bacteria, and the Ames test, which uses indicator strains of Salmonella typhimurium, sensitive to substances that can induce different types of mutation. Results: In the Inductest no decrease in survival fraction of bacteria and no increase in the formation of lysogenic induction were detected independently of the homeopathic potency employed. The same profile was obtained after the Ames test, with similar results to negative control. Conclusion: Afterwards, we can conclude that these samples are not able to induce DNA damage in the cells tested. So, the use of this medicine does not present any side effects related to mutagenesis and genotoxicity.(AU)

Evaluation of the genotoxic and mutagenic potentials of phytotherapic and homeopathic solutions of Euphorbia tirucalli Lineu (Aveloz) / Evaluation of the genotoxic and mutagenic potentials of phytotherapic and homeopathic solutions of Euphorbia tirucalli Lineu (Aveloz)

Introduction: Euphorbia tirucalli Lineu, commonly known as Aveloz, is a very common plant found in tropical regions [1]. The ingestion or contact with its latex causes symptoms such as vomiting and diarrhea, pallor, skin irritation, hepatotoxicity as well as carcinogenesis [2]. Moreover, the Aveloz latex is also responsible for a few important activities against some infectious and neoplastic diseases. Aveloz latex phytochemical composition may vary according to seasonal aspects and geographic location [3], and it is used either orally or topically in traditional medicine. Popularly known as an antitumoral agent (breast, prostate, lung, kidney), it is used not only in Brazil, but in several other countries. According to the literature, the latex could have a dual behaviour, activating or inhibiting tumoral events [3-6]. However, there are few reports discussing these mechanisms. Besides, the mutagenic and genotoxic potentials of phytochemical and homeopathic Aveloz have not yet been described. Several experimental methods have been used to evaluate the mutagenic and genotoxic effects, such as Inductest, the Ames test and the chromotest. Objective: This study aims to evaluate the genotoxic and mutagenic potentials of Aveloz latex and Aveloz phytotherapic and homeopathic solutions. Methodology: In this study, Aveloz 5 and 30cH are prepared according to Brazilian Homeopathic Pharmacopoeia [7], from Aveloz latex collected in the Center for Natural Products Research (NPPN) at the Universidade Federal do Rio de Janeiro [8]. The Aveloz phytochemical solution was prepared following the doses used in folk medicine: 2 drops diluted in 250ml of water and 2 drops diluted in 25 ml of water. All test solutions were submitted to the following methodologies: (a) Inductest: assesses the ability of physical or chemical agents to promote lysogenic induction as a response to DNA damage in lysogenic bacteria; (b) The Ames test: uses indicator strains of Salmonella typhimurium, which are sensitive to substances that can induce different types of mutation; (c) Chromotest: evaluates the genotoxicity of chemicals through the induction of the bacterial SOS system. Results: In the Inductest there was no decrease in bacterial survival fraction and no increase in lysogenic cycle. As measured by The Ames test and the chromotest no mutagenic or genotoxic potentials were detected. Discussion: The homeopathic and the phytochemical Aveloz solutions were unable to produce lysogenic induction or mutagenesis in bacterial cells and they were also unable to produce genotoxic effects, as measured by chromotest. Conclusion: Our results showed that no mutagenic or genotoxic damages were induced by all Aveloz preparations studied, thus we are led to believe that patients using Aveloz as a complementary therapy present no side effects in relation to mutagenesis and genotoxicity. (AU)

Evaluation of the genotoxic and mutagenic potentials of phytotherapic and homeopathic solutions of Euphorbia tirucalli Lineu (Aveloz) / Evaluation of the genotoxic and mutagenic potentials of phytotherapic and homeopathic solutions of Euphorbia tirucalli Lineu (Aveloz)

Introduction: Euphorbia tirucalli Lineu, commonly known as Aveloz, is a very common plant found in tropical regions [1]. The ingestion or contact with its latex causes symptoms such as vomiting and diarrhea, pallor, skin irritation, hepatotoxicity as well as carcinogenesis [2]. Moreover, the Aveloz latex is also responsible for a few important activities against some infectious and neoplastic diseases. Aveloz latex phytochemical composition may vary according to seasonal aspects and geographic location [3], and it is used either orally or topically in traditional medicine. Popularly known as an antitumoral agent (breast, prostate, lung, kidney), it is used not only in Brazil, but in several other countries. According to the literature, the latex could have a dual behaviour, activating or inhibiting tumoral events [3-6]. However, there are few reports discussing these mechanisms. Besides, the mutagenic and genotoxic potentials of phytochemical and homeopathic Aveloz have not yet been described. Several experimental methods have been used to evaluate the mutagenic and genotoxic effects, such as Inductest, the Ames test and the chromotest. Objective: This study aims to evaluate the genotoxic and mutagenic potentials of Aveloz latex and Aveloz phytotherapic and homeopathic solutions. Methodology: In this study, Aveloz 5 and 30cH are prepared according to Brazilian Homeopathic Pharmacopoeia [7], from Aveloz latex collected in the Center for Natural Products Research (NPPN) at the Universidade Federal do Rio de Janeiro [8]. The Aveloz phytochemical solution was prepared following the doses used in folk medicine: 2 drops diluted in 250ml of water and 2 drops diluted in 25 ml of water. All test solutions were submitted to the following methodologies: (a) Inductest: assesses the ability of physical or chemical agents to promote lysogenic induction as a response to DNA damage in lysogenic bacteria; (b) The Ames test: uses indicator strains of Salmonella typhimurium, which are sensitive to substances that can induce different types of mutation; (c) Chromotest: evaluates the genotoxicity of chemicals through the induction of the bacterial SOS system. Results: In the Inductest there was no decrease in bacterial survival fraction and no increase in lysogenic cycle. As measured by The Ames test and the chromotest no mutagenic or genotoxic potentials were detected. Discussion: The homeopathic and the phytochemical Aveloz solutions were unable to produce lysogenic induction or mutagenesis in bacterial cells and they were also unable to produce genotoxic effects, as measured by chromotest. Conclusion: Our results showed that no mutagenic or genotoxic damages were induced by all Aveloz preparations studied, thus we are led to believe that patients using Aveloz as a complementary therapy present no side effects in relation to mutagenesis and genotoxicity.