In vitro evaluation of Sporothrix brasiliensis biotherapic

Sporothrix brasiliensisis one of themost virulent zoonosis which affects animals and humans. This fungus is responsible for subcutaneous infection and its contamination is possible through trauma to the skin. Sporotrichosis is highly prevalent in feline. And Rio de Janeiro appears to have the highest occurrence of cases. Objectives: This study aims to evaluate the in vitroefficacy of Sporothrix brasiliensisbiotherapic, with and without an association to allopathic medicine commonly used in the treatment.Methodology: Conidiumcells of Sporothrix brasiliensiswill be cultured in Potato dextrose agar (PDA) for 5 to 7 days and yeast cells in Brain heart infusion (BHI) for 3 to 5 days. After incubation, the cells will be scraped with a drigalski handle and filtered using cells strainer into a tube and centrifuge for 5 minutes at 3000 RPM. The cells will be resuspended with Phosphate buffered saline (PBS), centrifuge again,and finally resuspended in PBS. After preparing the inocule, the microplates will be prepared. There will be 5groups in vitro. The first one will be the control group, only fungi. The second will be the treatment of fungi with homeopathic medicine (Sporothrix brasiliensis30DH). The third group will be the homeopathic medicine in association with itraconazole. The fourth will be the treatment with itraconazole only. And the last group will be the fungi with dynamized distilled water 30DH. Sporothrix brasiliensis30DHwill be prepared according to Brazilian Homeopathic Pharmacopeia. Results and discussions: The experiments are still in progress and the results will be analyzed through Analysis of variance (ANOVA) to determine statistically significant differences. Previous articles based on biotherapic treatments demonstrated successful results, so our research group is conducting these experiments to evaluate the effect in this model. Conclusion: Experiments will be made to verify the efficacy of biotherapic on sporotrichosis treatments.

Biotherapic 200dH is harmful to acute murine infection with Trypanosoma cruzi

Biotherapics employed to treat mice infected by Trypanosoma cruzi were carried out with encouraging results. The aim of this study was evaluated the effect of biotherapic of Trypanosoma cruzi 200dH, using two different schedules of treatment. Swiss male mice, aged 56 days-old were infected intraperitonealy with 1,400 blood trypomastigotes of Trypanosoma cruzi Y strain and were divided into groups: C.I.- infected animals, E.D. â€" Infected animals treated from the day 1 until the end of the experiment; 200dH S.D. â€" Infected animals treated on the day 1. Parasitological, clinical and immunological parameters were evaluated. The group of animals that received the medicine in a single dose presented higher value to total parasitaemia and lower value of pre patent period compared to control untreated group (p<0.05), as well as the number of amastigotes which was also higher for this group (S.D.) (p<0.05). Clinically, the S.D.group presented more stable temperature (p<0.05) but not presented another clinical difference among treatments. IL-6 and TNF-α presented similar dosage among treated groups as well as IL-4 and IL-10. IL-17A and INF-γ, presented highest values to S.D. group (p<0.05). All animals died until the 20th day of infection. The lack of improvement in clinical and parasitological parameters, the untimely death and the immunological imbalance display the harmful evolution of the experimental infection by T. cruzi using biotherapic 200dH. The results could be useful for homeopathic physicians. In human clinical use, the choice of dynamizations and treatment schedule should consider acute and chronic diseases to achieve the expected results. (AU)

Efeito de bioterápico na eosinofilia durante a SLMV experimental / Biotherapic effect in eosinophilia during experimental VLMS

O Toxocara canis (Tc) é um parasito pertencente ao filo Nematódeo que possui como hospedeiro definitivo os cães, O homem é hospedeiro paratênico e contamina-se acidentalmente ao ingerir ovos contendo larvas infectantes (L3) do parasito, as quais são liberadas e atravessam a mucosa intestinal, atingem a circulação, Durante este processo migratório, antígenos de excreção e secreção (TES) são liberados provocando intensa reação inflamatória, do tipo Th2, caracterizando a síndrome, denominada Larva Migrans Visceral (SLMV), As principais características desta doença crônica são as eosinofilias sanguínea e tecidual persistentes, Desse modo, torna-se importante a busca por terapias que contribuam com a redução dos quadros inflamatórios com intensa eosinofilia, Assim, o uso deste bioterápico, produzido a partir do extrato antigênico de ovos e larvas de (Tc), e seu efeito no recrutamento de leucócitos totais, células mononucleares e eosinófilos no sangue, para o espaço broncoalveolar e para a cavidade peritoneal de camundongos infectados pelo (Tc) foi investigado, Foram utilizados camundongos fêmeas (Swiss), divididos nos grupos: Controle (C), Infectado (Tc), Imunizado (Im+Tc) e Tratado (Tc+Bio), Os animais Tc, Im+Tc e Tc+Bio receberam 500 ovos/animal por gavagem, Posteriormente, os animais foram eutanasiados no 18º dia da infecção e o número das células nos compartimentos foi determinado, Os resultados obtidos demonstraram que, Im+Tc, assim como nos Tc+Bio tiveram redução significativa dessas células nos compartimentos analisados quando comparados grupo Tc, Assim, sugeriu-se que a bioterapia modulou negativamente o recrutamento de células inflamatórias, principalmente eosinófilos no sangue, pulmão e intestino demonstrando um potencial anti-inflamatório desse bioterápico na SLMV experimental...

The Toxocara canis (Tc) is a parasite that belongs to the nematode phylum and has dogs as definitive host. The men can be accidentally contaminated by ingesting eggs containing infective larvae of the parasite. These larvae, when ingested, pass through the intestinal mucosa, reach the portal circulation and migrate through different tissues of the host. During this process, excretory-secretory antigens (TES) are released causing an intense inflammatory reaction, the Th2 type, characterizing the syndrome, called Visceral Larva migrans (VLMS). The main features of this chronic disease are blood and tissue eosinophilias. Thus, it is important to search for therapies that may contribute to the reduction of inflammatory conditions with intense eosinophilia. In this study, we investigated the use of a biotherapic produced from the antigenic extract from eggs and larvae (Tc) and its effect on the recruitment of total leukocyte, mononuclear cells and eosinophils in blood, bronchoalveolar space and peritoneal cavity of mice infected with (Tc). Female mice (Swiss) were used divided in three groups: control (C), Infected (Tc) Immunized (Im + Tc) and Treaty (Tc + Bio). The animals Tc, Im + Tc and Tc + Bio received 500 eggs / animal by gavage. Subsequently, the animals were euthanized on day 18 after infection and the number of cells in the compartments was determined. Our results showed that, Im + Tc, as in Tc + Bio had reduced these cells in compartments analyzed compared to Tc group. Thus, it was suggested that the biotherapy negatively modulated the recruitment of inflammatory cells, particularly eosinophils in blood, lung and intestine demonstrating an anti-inflammatory potential of the biotherapic in the experimental VLMS...

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)

Respostas celulares e bioquímicas induzidas por bioterápicos preparados a partir de influenza A (H3N2) íntegro e influenza A (H3N2) inativado nas potências 12x e 30x na linhagem MDCK / Cellular and biochemical responses induced by Biotherapics prepared from intact influenza A (H3N2) and inactivated influenza A (H3N2) virus at 12x and 30x in the MDCK cells

Biotherapics are homeopathic remedies prepared from organic products that are chemically undefined and can be used for treatment of diseases like influenza. There are several classes of biotherapics and, among these, there are some called "living biotherapics" or "Roberto Costa?s Biotherapics". This study aimed to compare the cellular and biochemical effects of biotherapics prepared from intact influenza virus diluted in water and the one obtained from the same viral sample inactivated by ethanol 70% (v / v), both in the potencies of 12x and 30x. Transmission electron microscopy (TEM) analyses were performed on both preparations to assess the integrity of viral particles, which showed that ethanol 70% (v/v) induced a complete denaturation of viral particles. In contrast, the integrity of virus particles was preserved when water was used as the biotherapic solvent. Cellular and biochemical alterations induced by the preparations on MDCK cells were analyzed and compared with those induced by respective controls (water 30x-treated and untreated cells). Cellular viability analyzed by MTT method showed statistically significant differences (p <0.05) in MDCK cells treated with intact biotherapic for 5 (3 stimuli) and 30 (18 stimuli) days in comparison with untreated control. TEM analysis did not show significant cellular changes when the different experimental groups were compared. The enzymatic activity of phosphofructokinase 1 (PFK), an important enzyme in the glycolytic pathway, presented a statistically significant increase (p <0.05) after 30 days of treatment when compared to control groups. The results obtained suggest that inactivation of viral sample with ethanol 70% induces lysis and disruption of viral particles. In addition, preliminary results indicated that treatment with intact biotherapic seems to induce higher variations on MDCK cells responses when compared to inactivated-biotherapic-treated cells. Further analyses are ongoing, including scanning electron microscopy and quantification of the number of mitosis, in order to elucidate the mechanisms involved with biochemical and cellular responses induced by theses biotherapics.(AU)

Estabilidade Físico-Química e Microbiológica do Bioterápico Candida albicans RC em Variadas Potências / Physical-chemical and microbiological stability of biotherapy Candida albicans RC in different potencies

Oral Candidiasis is an opportunist fungal infection, with high incidence in HIV and immunosuppressed patients and Candida albicans is the most common causing agent. In some cases, it can evolve to resistant injuries to antifungal conventional therapy. According to Brazilian Homeopathic Pharmacopeia (BHP) [1], biotherapic medicines are prepared from chemically undefined biological products. Biotherapics created by Brazilian doctor Roberto Costa (RC) have a different homeopathic compounding technique, as its dynamization starts from the ethiologic agent of the illness in its alive form, which present higher capability to stimulate the host immunological system [2,3].Further experiments are being carried out in order to confirm the preliminary data obtained.(AU)

Tratamento com bioterápico Candida albicans influencia a adesão de fungos a células Ma104 in vitro / Treatment with Candida albicans biotherapic influences in vitro fungal adhesion to Ma-104 cells

Oral candidiasis is an opportunist fungal infection in humans, mainly caused by Candida albicans. It occurs when the host presents an imbalance in the immune system and Candida spp., normally found in human flora, become able to develop the infection [1]. This disease is very common in HIV patients, and in all individuals that present immunossupression, such as patients treated with chemotherapy. Considering this scenario, the development of new medicines to treat oral candidiasis is mandatory.These results showed that the biotherapic did not present any citotoxicity, but was able to modify the morphological aspects of Ma-104 cells. Additionally, the interaction between host cells and ethilogic agent is directly influenced by biotherapic treatment, suggesting a promising antifungal potential of this medicine.(AU)

Efeito de um bioterápico 7DH de Toxoplasma gondii em camundongos infectados com o protozoário / Effect of 7DH biotherapic of Toxoplasma gondii in mice infected with the protozoan

Toxoplasmosis is a zoonosis caused by Toxoplasma gondii worldwide distributed [1]. In both, men and animals, the infection with T. gondii can lead to important pathologies [2]. The study of alternative treatments is important to set new therapeutic protocols, especially for the prevention of congenital toxoplasmosis.Mice pre-infection treated with biotherapic 7DH, presented bigger clinical alterations, which were measured visually and statistically compared to the control group. There was a biological effect of the biotherapic, with an increase in the number of cysts compared to the control group, without statistical significance. The group 7DH showed a significant reduction of intraocular pressure and fundoscopic analysis showed a larger number of animals without ocular changes, without statistical significance. The sample size should be reevaluated for better data interpretation and decision on the effects of the biotherapic 7DH T. gondii.(AU)

Influência da idade e esquema de tratamento na parasitemia de camundongos infectados pelo Trypanosoma cruzi tratados com bioterápico de alta potência / Influence of age and ways of treatment in the parasitemia in mice infected with Trypanosoma cruzi treated with high potency biotherapy

ABSTRACTIntroduction: The infection of mice by Trypanosoma cruzi is well known, making this a good model for understanding the effect of highly diluted medications. Mice of different ages show different responses to biotherapic T. cruzi [1]. Other data from our laboratory using biotherapic treatment at low potencies show that long lasting treatment has a better effect in mice infected with T. cruzi. However, the use of high potency biotherapics in mice of different ages infected with T. cruzi has not been analysed yet.Conclusion: The age and the ways of treatment used are important factors to be considered when using a highly diluted medication. The clinical use of these results in humans, should take into consideration the allometric system of medication dosage which takes into account the metabolic rate of each organism.(AU)

Introdução: A infecção murina pelo Trypanosoma cruzi é bem conhecida, fazendo deste um bom modelo para o entendimento do efeito de medicamentos ultradiluídos. Camundongos de diferentes idades mostram diferentes respostas a bioterápico de T. cruzi [1]. Outros dados do nosso laboratório utilizando tratamento com bioterápico em baixas potências mostram que o tratamento prolongado exerce melhor efeito em camundongos infectados pelo T. cruzi. No entanto, a utilização de bioterápicos em alta potência em camundongos de diferentes idades infectados pelo T. cruzi ainda não foi explorada.Conclusão: A idade e o esquema de tratamento utilizado são fatores importantes a serem considerados na utilização de medicamento ultradiluído. A utilização clínica destes resultados em humanos, deve considerar o sistema alométrico de dosagem de medicamentos que leva em conta a taxa metabólica de cada organismo.(AU)

Bioterápico 17DH T. cruzi em uso prolongado melhora clinicamente camundongos infectados pelo Trypanosoma cruzi / Biotherapic T. cruzi 17DH when continuously used clinically improves mice infected with Trypanosoma cruzi.

Introduction: In Trypanosoma cruzi infection, the pathogenesis is the result of a rupture in the host - parasite relationship [1]. This rupture is related to the imbalance of the vital force of the host, expressed through signs and symptoms, defined by Hahnemann (1995)[2] as being the source of the disease. There is no research in the literature about the clinical evolution of mice experimentally infected with T. cruzi and treated in different ways using biotherapic. Therefore, this is an area to be studied in the future.Conclusion: The use of biotherapic T. cruzi 17DH for a long period causes clinical improvement of the infected mice with Trypanosoma cruzi. The clinical use of these results in human beings should consider the allometric medicine dosage which takes into account the metabolic rate of each organism(AU)

Introdução: Na infecção pelo Trypanosoma cruzi, a patogenia é resultado do rompimento do equilíbrio da relação parasito - hospedeiro (Tafuri, 1987), que está relacionada com o desequilíbrio da força vital do hospedeiro, expressando-se através de sinais e sintomas, definido por Hahnemann (2007), como sendo a origem da doença. Não existe na literatura trabalhos que abordem a evolução clínica de camundongos experimentalmente infectados pelo T. cruzi e tratados com diferentes esquemas de tratamento utilizando bioterápico. Sendo assim, é necessária a realização de estudos com este objetivo.Conclusão: O uso prolongado do bioterápico 17DH T. cruzi melhora clinicamente camundongos infectados pelo T. cruzi. A utilização clínica destes resultados em humanos, deve considerar o sistema alométrico de dosagem de medicamentos que leva em conta a taxa metabólica de cada organismo.(AU)

Avaliação da resposta imune e comportamental de camundongos BALB/c após tratamento com soluções homeopáticas de H3N2 / Evaluation of BALB/c mice behavior and relations to the immune response after treatment with H3N2 homeopathic solutions

In Brazil, homeopathic medicines are prepared according to the Homeopathic Pharmacopeia, regulated by ANVISA. Among several categories of medicines, there is the biotherapic group, which is prepared from etiologic agents. In this study, we developed a biotherapic from influenza A virus, aiming the influenza infection prevention. Influenza is a disease that affects thousands of people worldwide every year, with an important economic impact, what motivates the development of new low cost therapies. The H3N2 biotherapic developed in this study was administered to Balb/c mice to evaluate their immune response to viral specific antigens and behavior (homeopathic proving). Sixty-two 4 weeks old Balb/c mice were divided into five experimental groups (n=14 per group), after approval by the Ethics Committee of Animal Use (Protocol DFBCICB 037) and stimulated daily, blindly, with 1% (v/v) different homeopathic medicines, for a maximum period of 42 days. The tested medicines were: biotherapic 30x prepared from inactivated influenza A virus; biotherapic 30x prepared with infectious influenza A virus; and thymulin 5cH, a thymus hormone. The two control groups were treated with water 30x and nothing (baseline group). After 21 days of treatment, half of the animals from each group was challenged subcutaneously with the viral hemagglutinin antigen (7 g / 200 L) and monitored by 21 days further, to evaluate the humoral immune response and general behavior, using an open device. The remaining animals were evaluated by the same behavioral tests at the end of the first 21 days, as an attempt to define the proving features. After euthanasia, all animals were autopsied and the spleen, lungs, heart and mediastine lymph nodes were weighed. Histometry of the spleen follicles was also made. Histopathological and behavioral analyses showed absence of behavioral effects, however, there was increase of spleen lymphoid follicles diameter in immunized animals treated with thymulin and with the biotherapic prepared from infectious influenza A, when compared to the control group. This experiment is being repeated using flow cytometry to complete the analysis and confirm the results.(AU)

Medicamentos homeopáticos são preparados de acordo com a farmacotécnica homeopática regulamentada pela ANVISA. Dentre as várias categorias destes medicamentos, destaca-se o grupo dos bioterápicos, medicamentos que são preparados a partir do próprio agente etiológico. No presente estudo, foi desenvolvido um bioterápico a partir do vírus influenza A, visando a profilaxia da gripe. A gripe é uma doença que atinge milhares de pessoas anualmente em todo o mundo e o desenvolvimento de novas terapias para esta doença vem sendo estimulado com frequência. O bioterápico desenvolvido foi administrado a camundongos do tipo Balb/c para avaliação da resposta imune e comportamental. Para tanto, sessenta e dois camundongos Balb/c com 4 semanas de vida foram separados em cinco grupos experimentais, após aprovação pelo Comitê de Ética de Uso de Animais (Protocolo DFBCICB 037) e estimulados diariamente, de maneira cega, por diferentes soluções homeopáticas, na concentração de 1% (V/V), durante um prazo máximo de 42 dias. Três medicamentos homeopáticos foram testados: bioterápico contendo o vírus influenza A inativado 30DH; bioterápico contendo o vírus influenza A íntegro 30DH; timulina 5CH. Um grupo controle foi tratado com água 30x e o outro não recebeu tratamento. Após 21 dias de tratamento, metade dos animais de cada grupo (31 animais) foi desafiada, por via subcutânea, com o antígeno viral hemaglutinina na concentração de 7 g/ 200L e acompanhados por mais 21 dias para avaliação da resposta imune humoral e do comportamento, pela técnica do campo aberto. Os animais restantes foram submetidos aos mesmos testes ao final dos primeiros 21 dias de tratamento, antes do desafio antigênico. Após a eutanásia, todos os animais foram necropsiados e o baço, o pulmão, o coração e o linfonodo mediastínico foram colhidos para análise de peso e histometria do baço. As análises histopatológica e comportamental mostraram a ausência de efeitos patogenéticos perceptíveis neste modelo experimental, mas houve aumento da reatividade dos folículos linfóides do baço nos animais desafiados antigenicamente e tratados com bioterápico de influenza A íntegro e timulina, em relação ao grupo controle. Este experimento está sendo repetido(AU)

Bioterápico 17DH de Trypanosoma cruzi aumenta a apoptose em camundongos experimentalmente infectados / Biotherapic of Trypanosoma cruzi 17d increases apoptosis in experimentally infected mice

Introduction: the mechanism of action of ultradiluted medicines has not yet been established[1,3]. Many basic research studies have focused on isopathic models using in vitro or in vivo designs [4,5]. Recent studies indicate that an ultradiluted (isopathic) antigen can transfer signals to the immune system and modulate its response when an organism is challenged against this same antigen [6]. Some studies on experimental infection of mice by T. cruzi identified apoptotic cells and showed that the increase of their number is associated with an increase also in the number of parasites in the blood of the infected animals, while blockage of apoptosis can be the target of therapeutic intervention [7,8].Conclusion: these results show that apoptosis is increased in animals treated with biotherapic of T. cruzi 17d.n(AU)

Introdução: O mecanismo de ação de medicamentos ultradiluídos ainda não está elucidado [1-3]. Muitos estudos em pesquisas básicas concentraram-se nos modelos de isopatia, utilizando protocolos in vivo ou in vitro [4,5]. Estudos recentes relatam que um antígeno ultradiluído (isopático) pode transferir sinais para o sistema imunológico e modular a sua resposta quando o organismo é desafiado contra este antígeno [6]. Outros trabalhos mostraram que na infecção experimental de camundongos pelo T. cruzi foram detectadas células apoptóticas e que um aumento das células apoptóticas está relacionado ao aumento da parasitemia em animais infectados e que o bloqueio da apoptose pode ser alvo de intervenção terapêutica [7,8].Conclusão: A apoptose está aumentada em animais tratados com bioterápico 17DH de T. cruzi e infectados pelo protozoário.(AU)

Bioterápico 17DH de Trypanosoma cruzi diminui alterações histopatológicas em camundongos infectados pelo protozoário / Biotherapic of Trypanosoma cruzi 17x controlled histopathological alterations in mice infected by this protozoon

Introduction: about 10 million people worldwide suffer from Chagas? disease [1]. The World Health Organization (WHO) has explicitly acknowledged the significance of this condition and supports the use of Complementary and Alternative Medicine by health systems integrated with conventional treatments. Even so, one century after its discovery it still represents a global challenge [1,2]. Biotherapics are ultradiluted medicines and the infection of mice by Trypanosoma cruzi is an excellent model to understand their effect [3,4]. At 8 weeks, mice are physiologically more developed than at age 4 weeks, including a more competent immune system [5].Conclusion: there is a difference in the effect of the ultradiluted medicine between mice 4- and 8-week old. Eight-week old animals treated with biotherapic exhibited lower tissue parasitism, which is the opposite of what was observed in 4-week old animals.(AU)

Cell alterations induced by a biotherapic for influenza / Cell alterations induced by a biotherapic for influenza

Introduction: Influenza viruses have been responsible for highly contagious acute respiratory illnesses with high mortality, mainly in the elderly, which encourages the development of new drugs for the treatment of human flu. The biotherapics are medicines prepared from biological products, which are not chemically defined. They are compounded following the homeopathic procedures indicated for infectious diseases with known etiology [1]. Aim: The purpose of the present study is to verify cellular alterations induced by a biotherapic prepared from the infectious influenza A virus. Methodology: This biotherapic was prepared for this study in the homeopathic potency of 30X according to the Brazilian Homeopathic Pharmacopeia [2]. The concentration of 10% was not cytotoxic to cells, as verified by neutral red assay. The cellular alterations observed in MDCK cells were analyzed by optical microscopy for the quantification of mitosis, nucleoli and lipid bodies. The mitochondrial activity was assessed by MTT assay and the phosphosfructokinase-1 (PFK-1) enzyme activity was analyzed on the MDCK cells treated for 5, 10 and 30 days. Macrophages J778.G8 were treated with this biotherapic to evaluate the immunostimulatory cytokine release. Results: The cellular alterations observed in MDCK cells were verified by optical microscopy. The number of lipid bodies present in MDCK cells stimulated for 10 days was significantly lower (p <0.05) when compared to controls. The biotherapic significantly increased (p <0.05) the number of mitosis and the mitochondrial activity of MDCK cells stimulated for 10 and 30 days. These changes were confirmed by a significant reduction (p <0.05) on the PFK-1 activity. These results suggest that the biotherapic was able to activate the Krebs cycle and pentosephosphate metabolism to the generation of amino acids and nucleotides, situations common to cells whose rate of mitosis is increased. The quantification of immunostimulatory cytokines by macrophages J774.G8 indicated that the tumor necrosis factor (TNF-?) production was higher (p <0.05) in the supernatant of the macrophages pre-treated with this biotherapic and infected with influenza virus, suggesting an activation of the macrophages by this biotherapic. Conclusion: This biotherapic is able to induce some cellular alterations, which show strong evidence that it might be a promising option for the human flu. New experiments are being developed to understand the mechanisms of action of this biotherapic.(AU)

Cell alterations induced by a biotherapic for influenza / Cell alterations induced by a biotherapic for influenza

Introduction: Influenza viruses have been responsible for highly contagious acute respiratory illnesses with high mortality, mainly in the elderly, which encourages the development of new drugs for the treatment of human flu. The biotherapics are medicines prepared from biological products, which are not chemically defined. They are compounded following the homeopathic procedures indicated for infectious diseases with known etiology [1]. Aim: The purpose of the present study is to verify cellular alterations induced by a biotherapic prepared from the infectious influenza A virus. Methodology: This biotherapic was prepared for this study in the homeopathic potency of 30X according to the Brazilian Homeopathic Pharmacopeia [2]. The concentration of 10% was not cytotoxic to cells, as verified by neutral red assay. The cellular alterations observed in MDCK cells were analyzed by optical microscopy for the quantification of mitosis, nucleoli and lipid bodies. The mitochondrial activity was assessed by MTT assay and the phosphosfructokinase-1 (PFK-1) enzyme activity was analyzed on the MDCK cells treated for 5, 10 and 30 days. Macrophages J778.G8 were treated with this biotherapic to evaluate the immunostimulatory cytokine release. Results: The cellular alterations observed in MDCK cells were verified by optical microscopy. The number of lipid bodies present in MDCK cells stimulated for 10 days was significantly lower (p <0.05) when compared to controls. The biotherapic significantly increased (p <0.05) the number of mitosis and the mitochondrial activity of MDCK cells stimulated for 10 and 30 days. These changes were confirmed by a significant reduction (p <0.05) on the PFK-1 activity. These results suggest that the biotherapic was able to activate the Krebs cycle and pentosephosphate metabolism to the generation of amino acids and nucleotides, situations common to cells whose rate of mitosis is increased. The quantification of immunostimulatory cytokines by macrophages J774.G8 indicated that the tumor necrosis factor (TNF-?) production was higher (p <0.05) in the supernatant of the macrophages pre-treated with this biotherapic and infected with influenza virus, suggesting an activation of the macrophages by this biotherapic. Conclusion: This biotherapic is able to induce some cellular alterations, which show strong evidence that it might be a promising option for the human flu. New experiments are being developed to understand the mechanisms of action of this biotherapic.

Efficiency of tick biotherapic on the control of infestation by Rhipicephalus (Boophilus) microplus in Dutch dairy cows / Eficiência de bioterápico de carrapato no controle da infestação por rhipicephalus (boophilus) microplus em vacas leiteiras de raça holandesa / Eficiencia de bioterápico de garrapata para control de la infestación por Rhipicephalus (Boophilus) microplus en vacas lecheras de raza holandesa

Background: cattle tick Rhipicephalus (Boophilus) microplus poses serious problems for farmers in Brazil, especially because the parasite easily develops resistance to pesticide agents. For this reason, together with other factors including environmental, human and animal contamination and costs, alternative approaches have been sought for. Aims: this study sough to evaluate the efficiency of a tick biotherapic on tick-infested cows. Methods: 34 dairy Dutch cows were divided in 2 groups: one group received 100g/day of mineral salt supplement impregnated with tick biotherapic 12cH for 6 months, and then in alternate days with tick biotherapic 30cH to complete 28 months of treatment; the other group (control) received only the mineral salt supplement. After 28 months of treatment, engorged Rhipicephalus (boophilus) microplus females were collected in both groups, counted and weighed; in vitro tests were carried out to assess mass of ticks; egg mass; egg-hatching rate; and reproductive efficiency. Results: There was significant difference between both groups for all parameters evaluated; tick-mass (p = 0.0008); egg mass (p=0.0044); egg-hatching rate (p= 0.0017); and reproductive efficiency (p = 0.0044). Conclusion: treatment with tick biotherapic significantly decreased the mass of engorged females, deposition and hatching rate of eggs, resulting consequently in the decrease of the reproductive efficiency of ticks.(AU)

Introdução: os carrapatos do gado Rhipicephalus (Boophilus) microplus representam um sério problema para os criadores brasileiros, especialmente porque desenvolvem rapidamente resistência aos pesticidas. Por isso, junto a outros fatores incluindo contaminação ambiental, humana e animal, assim como os custos, têm sido procuradas abordagens alternativas. Objetivos: avaliar a eficiência de um bioterápico desenvolvido a partir de carrapatos em vacas infestadas. Métodos: 34 vacas leiteiras de raça holandesa foram divididas em 2 grupos; um recebeu 100 mg/dia de suplemento salino mineral impregnado com o bioterápico 12cH durante 6 meses e após 30cH em dias alternos até completar 28 meses de tratamento; o outro grupo (controle) recebeu só o suplemento mineral. Após 28 meses de tratamento foram coletadas fêmeas ingurgitadas do carrapato em ambos os grupos, contadas e pesadas, e foram realizados testes in vitro para determinar a massa de parasitas; massa de ovos; taxa de eclosão dos ovos; e eficiência reprodutiva. Resultados: houve diferença significativa entre ambos os grupos em todos os parâmetros avaliados ? massa de parasitas (p=0,0008); massa de ovos (p=0,0044); taxa de eclosão de ovos (p=0,0017) e eficácia reprodutiva (0,0044). Conclusão: o tratamento com bioterápico de carrapato diminuiu significativamente a massa de fêmeas ingurgitadas e a deposição e taxa de eclosão, resultando, consequentemente, na diminuição da eficiência reprodutiva dos carrapatos.(AU)

Introducción: la garrapata del ganado Rhipicephalus (Boophilus) microplus produce serios problemas a los hacenderos brasileños especialmente porque desarrolla rápidamente resistencia a los agentes pesticidas. Por este motivo, asociado a otros factores como contaminación ambiental, humana y animal y los costos, se buscan abordajes alternativos. Objetivos: este estudio buscó evaluar la eficiencia de un bioterápico preparado de garrapatas en vacas infestadas. Métodos: 34 vacas lecheras de raza holandesa fueron divididas en 2 grupos; uno recibió 100 mg/día de suplemento mineral impregnado con bioterápico 12cH durante 6 meses y después, en días alternados hasta completar 28 meses de tratamiento; el otro recibió apenas el suplemento mineral (control). Después de 28 meses de tratamiento, garrapatas hembra ingurgitadas fueron recogidas en ambos grupos, contadas y pesadas; fueron realizados pruebas in vitro para medir la masa de garrapatas; la masa de huevos, la tasa de eclosión de huevos y la eficiencia reproductiva. Resultados: hubo diferencias significativas entre los 2 grupos en todos los parámetros evaluados ? masa de garrapatas (p=0,0008), masa de huevos (p=0,0044), tasa de eclosión de huevos (p=0,0017) y eficiencia reproductiva (p=0,0044). Conclusión: el tratamiento con bioterápico de garrapata redujo significativamente la masa de hembras ingurgitadas y la deposición y eclosión de huevos, resultando en disminución de la eficacia reproductiva de las garrapatas.(AU)

Efficiency of tick biotherapic on the control of infestation by rhipicephalus (boophilus) microplus in dutch dairy cows / Eficiência de bioterápico de carrapato no controle da infestação por rhipicephalus (boophilus) microplus em vacas leiteiras de raça holandesa / Eficiencia de bioterápico de garrapata para control de la infestación por Rhipicephalus (Boophilus) microplus en vacas lecheras de raza holandesa

Background: cattle tick Rhipicephalus (Boophilus) microplus poses serious problems for farmers in Brazil, especially because the parasite easily develops resistance to pesticide agents. For this reason, together with other factors including environmental, human and animal contamination and costs, alternative approaches have been sought for. Aims: this study sough to evaluate the efficiency of a tick biotherapic on tick-infested cows. Methods: 34 dairy Dutch cows were divided in 2 groups: one group received 100g/day of mineral salt supplement impregnated with tick biotherapic 12cH for 6 months, and then in alternate days with tick biotherapic 30cH to complete 28 months of treatment; the other group (control) received only the mineral salt supplement. After 28 months of treatment, engorged Rhipicephalus (boophilus) microplus females were collected in both groups, counted and weighed; in vitro tests were carried out to assess mass of ticks; egg mass; egg-hatching rate; and reproductive efficiency. Results: There was significant difference between both groups for all parameters evaluated; tick-mass (p = 0.0008); egg mass (p=0.0044); egg-hatching rate (p= 0.0017); and reproductive efficiency (p = 0.0044). Conclusion: treatment with tick biotherapic significantly decreased the mass of engorged females, deposition and hatching rate of eggs, resulting consequently in the decrease of the reproductive efficiency of ticks.

Introdução: os carrapatos do gado Rhipicephalus (Boophilus) microplus representam um sério problema para os criadores brasileiros, especialmente porque desenvolvem rapidamente resistência aos pesticidas. Por isso, junto a outros fatores incluindo contaminação ambiental, humana e animal, assim como os custos, têm sido procuradas abordagens alternativas. Objetivos: avaliar a eficiência de um bioterápico desenvolvido a partir de carrapatos em vacas infestadas. Métodos: 34 vacas leiteiras de raça holandesa foram divididas em 2 grupos; um recebeu 100 mg/dia de suplemento salino mineral impregnado com o bioterápico 12cH durante 6 meses e após 30cH em dias alternos até completar 28 meses de tratamento; o outro grupo (controle) recebeu só o suplemento mineral. Após 28 meses de tratamento foram coletadas fêmeas ingurgitadas do carrapato em ambos os grupos, contadas e pesadas, e foram realizados testes in vitro para determinar a massa de parasitas; massa de ovos; taxa de eclosão dos ovos; e eficiência reprodutiva. Resultados: houve diferença significativa entre ambos os grupos em todos os parâmetros avaliados ? massa de parasitas (p=0,0008); massa de ovos (p=0,0044); taxa de eclosão de ovos (p=0,0017) e eficácia reprodutiva (0,0044). Conclusão: o tratamento com bioterápico de carrapato diminuiu significativamente a massa de fêmeas ingurgitadas e a deposição e taxa de eclosão, resultando, consequentemente, na diminuição da eficiência reprodutiva dos carrapatos.

Introducción: la garrapata del ganado Rhipicephalus (Boophilus) microplus produce serios problemas a los hacenderos brasileños especialmente porque desarrolla rápidamente resistencia a los agentes pesticidas. Por este motivo, asociado a otros factores como contaminación ambiental, humana y animal y los costos, se buscan abordajes alternativos. Objetivos: este estudio buscó evaluar la eficiencia de un bioterápico preparado de garrapatas en vacas infestadas. Métodos: 34 vacas lecheras de raza holandesa fueron divididas en 2 grupos; uno recibió 100 mg/día de suplemento mineral impregnado con bioterápico 12cH durante 6 meses y después, en días alternados hasta completar 28 meses de tratamiento; el otro recibió apenas el suplemento mineral (control). Después de 28 meses de tratamiento, garrapatas hembra ingurgitadas fueron recogidas en ambos grupos, contadas y pesadas; fueron realizados pruebas in vitro para medir la masa de garrapatas; la masa de huevos, la tasa de eclosión de huevos y la eficiencia reproductiva. Resultados: hubo diferencias significativas entre los 2 grupos en todos los parámetros evaluados ? masa de garrapatas (p=0,0008), masa de huevos (p=0,0044), tasa de eclosión de huevos (p=0,0017) y eficiencia reproductiva (p=0,0044). Conclusión: el tratamiento con bioterápico de garrapata redujo significativamente la masa de hembras ingurgitadas y la deposición y eclosión de huevos, resultando en disminución de la eficacia reproductiva de las garrapatas.

Influência do grupo genético, condição sexual e tratamento antiparasitário nas medidas de área de olho do lombo e espessura de gordura in vivo e na carcaça de bovinos de corte / Influence of breed, gender condition, and anti-parasitic treatment on the measurements of in vivo rib eye area and fat thickness and on the carcass of beef cattle

Foram estudados 48 bovinos machos oriundos de inseminação artificial, criados em pasto, sendo 24 (12 Nelore e 12 F1 ½ Red Angus-Nelore ) tratados com antiparasitários alopáticos e 24 (mesmo número de puros e cruzados) tratados com o antiparasitário bioterápico Fator C&MC. Os animais foram desmamados aos oito meses, metade de cada subgrupo genético (6) foi castrado aos 13 meses e todos abatidos aos 32 meses, com o objetivo de verificar a influência do tratamento antiparasitário, do grupo genético e da condição sexual sobre as medidas de área de olho de lombo (AOL) e espessura de gordura de lombo (EGL). Usaram-se medidas de ultrassonografia no animal vivo (AOLU e EGLU) e na carcaça, plástico quadriculado e paquímetro (AOLC e EGLC). Os animais F1, os inteiros e os tratados com alopatia apresentaram peso vivo maior quando comparados aos Nelores, castrados e tratados com bioterápicos. Não houve diferença da AOLU e AOLC entre os grupos genéticos. EGLC foi mais alta nos cruzados. Os animais inteiros apresentaram AOLU e AOLC maiores que os castrados, e EGLU e EGLC menores. Foram altamente significativos os coeficientes de correlação entre as medidas por ultrassom e na carcaça para área de olho de lombo (0,87) e espessura de gordura do lombo (0,95).

Forty-eight male bovines, products of artificial insemination and raised on pasture, were studied, being 24 (12 Nelore, 12 F1 ½ Nelore ½ Red Angus) treated with allopathic antiparasitic drugs and 24 (same number of pure and crossbred) treated with a biotherapic antiparasitic drug Factor C&MC. Animals were weaned at eight months of age and half of each genetic subgroup (six) was castrated at 13 months of age. All animals were slaughtered at 32 months of age, in an attempt to evaluate the influence of antiparasitic treatment, genetic group, and gender condition in the measurements of rib eye area (AOL) and fat thickness (EGL) of loin. Measurements of ultrasonography were used for live animals (AOLU and EGLU), whereas a direct plastic grid reading of the eye muscle and a pachymeter (AOLC and EGLC) were used for carcasses. F1 animals, non-castrated, and those treated with allopathic drugs showed higher live weight when compared with Nelore, castrated, and biotherapic treated animals. There were no differences between genetic groups of AOLU and AOLC. EGLC was higher in crossbred animals. Non-castrated animals showed higher AOLU and AOLC when compared with castrated animals, and lower EGLU and EGLC. Correlation coefficients for ultrasound and carcass measurements were highly significant for rib eye area (0.87) and fat thickness (0.95).