Structural and thermal analyses of zinc and lactose in homeopathic triturated systems.

BACKGROUND: A series of different experimental approaches was applied in Zincum metallicum (Zinc met.) samples and lactose controls. Experiments were designed to elucidate the effect of zinc trituration and dynamization on physicochemical properties of homeopathic formulations, using lactose as excipient. METHODS: Zinc met. potencies (Zinc met 1-3c) were triturated and dynamized using lactose as excipient, according to Brazilian Homeopathic Pharmacopoeia. Lactose samples (LAC 1-3c) were also prepared following the same protocol and used as controls. The samples were analyzed structurally by Atomic Absorption Spectroscopy (AAS), X-ray Diffraction (XRD), Transmission Electron Microscopy (TEM) with Energy Dispersive X-ray Spectroscopy (EDX) and Scanning Electron Microscopy (SEM), and thermodynamically by Thermogravimetry (TG) and Differential Scanning Calorimetry (DSC). RESULTS: AAS analysis detected 97.0 % of zinc in the raw material, 0.75 % (Zinc met 1c) and 0.02% (Zinc met 2c). XRD analysis showed that inter-atomic crystalline spacing of lactose was not modified by dynamization. Amorphous and crystalline lactose spheres and particles, respectively, were observed by TEM in all samples, with mean size from 200 to 800 nm. EDX obtained with TEM identified zinc presence throughout the amorphous matter but individualized zinc particles were not observed. SEM images obtained from dynamized samples (LAC 1c and Zinc met 1c) with electron backscattering could not identify zinc metal grains. The dynamization process induced Derivatives of Thermal Gravimetric (DTg) peak modification, which was previously centered near 158°C to lactose, to a range from 140 to 170°C, suggesting the dynamization process modifies the temperature range of water aggregation. Thermal phenomena were analyzed and visualized by Analysis of Variance (ANOVA) and Principal Component Analysis (PCA) statistics. Both indicated that fusion enthalpy of dynamized samples (DynLAC 1-3c; DynZn 1-3c) increased 30.68 J/g in comparison to non-dynamized lactose (LAC; p < 0.05). CONCLUSIONS: Our results suggested no structural changes due to the trituration and dynamization process. However, TG and DSC analyses permit the differentiation of dynamized and non-dynamized groups, suggesting the dynamization process induced a significant increase in the degradation heat. These results call for further calorimetric studies with other homeopathic dilutions and other methodologies, to better understand the dynamics of these systems.

Homeopathic treatments modify inflammation but not behavioral response to influenza antigen challenge in BALB/c mice.

BACKGROUND: Influenza affects thousands of people worldwide every year, motivating the development of new therapies. In this work, the effects of two homeopathic preparations (influenza biotherapies and thymulin) were chosen following two different rationales: isotherapy and endo-isotherapy models. The homeopathic effects were evaluated individually considering the inflammatory and behavioral responses against influenza virus antigen were studied in BALB/c mice. METHODS: Male adult mice were treated orally and blindly for 21 days with highly diluted influenza virus or with thymulin, and were divided in two sets of experiments. The first series of experiments aimed to describe their behavior, using an open field (OF) device. In the second series, mice were challenged subcutaneously with influenza hemagglutinin antigen (7 µg/200 µl) at day 21. At day 42, behavior and inflammation response were evaluated. RESULTS: No behavioral changes were seen in OF tests at any time point after treatments. Flow cytometry and morphometry revealed significant changes in T and B cell balance after influenza antigen challenge, varying according to treatment. CONCLUSION: The results show that both homeopathic treatments induced subtle changes in acquired immune anti-viral response regulation. A deeper understanding of the mechanism could elucidate their possible use in influenza epidemiological situations.

Direct electric current treatment modifies mitochondrial function and lipid body content in the A549 cancer cell line.

Electrochemical therapy (EChT) entails treatment of solid tumors with direct electric current (DC). This work evaluated the specific effects of anodic flow generated by DC on biochemical and metabolic features of the A549 human lung cancer cell line. Apoptosis was evaluated on the basis of caspase-3 activity and mitochondrial transmembrane potential dissipation. Cell morphology was analyzed using transmission electron microscopy, and lipid droplets were studied through morphometric analysis and X-ray qualitative elemental microanalysis. High-resolution respirometry was used to assess mitochondrial respiratory parameters. Results indicated A549 viability decreased in a dose-dependent manner with a prominent drop between 18 and 24h after treatment (p<0.001), together with a two-fold increase in caspase-3 activity. AF-treatment induced a significantly increase (p<0.01) in the cell number with disrupted mitochondrial transmembrane potential. Furthermore, treated cells demonstrated important ultrastructural mitochondria damage and a three-fold increase in the cytoplasmic lipid bodies' number, quantified by morphometrical analyses. Conversely, 24h after treatment, the cells presented a two-fold increase of residual oxygen consumption, accounting for 45.3% of basal oxygen consumption. These results show remarkable alterations promoted by anodic flow on human lung cancer cells which are possibly involved with the antitumoral effects of EChT.

Homeopathic medicines for prevention of influenza and acute respiratory tract infections in children: blind, randomized, placebo-controlled clinical trial.

BACKGROUND: Influenza and its complications are common at all ages, especially in children. Vaccines and anti-influenza drugs aim to prevent it. Preventative approaches with favorable risk profiles should be considered for flu, particularly since the evidence of the efficacy of anti-viral drugs is debated. METHODS: This pragmatic clinical trial was conducted in the Brazilian Public Health System in Petrópolis (BPHSP) with children aged from 1 to 5 years old. The medications used were mainly selected based on in vitro experiments (InfluBio), and in successful qualitative clinical experiences (Homeopathic Complex). Following informed parental consent, subjects were randomly distributed, in a blind manner, to three experimental groups: Homeopathic Complex, Placebo, and InfluBio. BPHSP health agents collected flu and acute respiratory infection symptomatic episodes monthly following the established protocol. The number of these episodes was registered in one year (2009-2010). RESULTS: Out of the 600 children recruited, 445 (74.17%) completed the study (149: Homeopathic complex; 151: Placebo; 145: InfluBio). The number of flu and acute respiratory infection symptomatic episodes detected in this clinical trial was low; however, it was different between homeopathic groups and placebo (p < 0.001). In the first year post-intervention, 46/151 (30.5%) of children in the placebo group developed 3 or more flu and acute respiratory infection episodes, while there was no episode in the group of 149 children who used Homeopathic Complex, and only 1 episode in the group of 145 (1%) children who received InfluBio. CONCLUSION: These results suggested that the use of homeopathic medicines minimized the number of flu and acute respiratory infection symptomatic episodes in children, signalizing that the homeopathic prophylactic potential should be investigated in further studies.

H3N2 homeopathic influenza virus solution modifies cellular and biochemical aspects of MDCK and J774G8 cell lines.

BACKGROUND: Influenza viruses cause highly contagious acute respiratory illnesses with significant mortality, especially among young children, elderly people, and individuals with serious medical conditions. This encourages the development of new treatments for human flu. Biotherapies are diluted solutions prepared from biological products compounded following homeopathic procedures. OBJECTIVES: To develop a biotherapy prepared from the infectious influenza A virus (A/Aichi/2/68 H3N2) and to verify its in vitro response. METHODS: The ultradiluted influenza virus solution was prepared in the homeopathic dilution 30dH, it was termed Influenzinum RC. The cellular alterations induced by this preparation were analyzed by optical and electron microscopy, MTT and neutral red assays. Glycolytic metabolism (PFK-1) was studied by spectrophotometric assay. Additionally, the production of tumor necrosis factor-α (TNF-α) by J774.G8 macrophage cells was quantified by ELISA before and after infection with H3N2 influenza virus and treatment. RESULTS: Influenzinum RC did not cause cytotoxic effects but induced morphological alterations in Madin-Darby canine kidney (MDCK) cells. After 30 days, a significant increase (p < 0.05) in mitosis rate was detected compared to control. MDCK mitochondrial activity was changed after treatment for 10 and 30 days. Treatment significantly diminished (p < 0.05) PFK-1 activity. TNF-α in biotherapy-stimulated J774.G8 macrophages indicated a significant (p < 0.05) increase in this cytokine when the cell supernatant was analyzed. CONCLUSION: Influenzinum RC altered cellular and biochemical features of MDCK and J774G8 cells.

Bioterápicos de Candida albicans induzem alterações nos índices de adesão e na respiração celular / Biotherapics of Candida albicans induce changes in the levels of adhesion and in cellular respiration

Introdução: O gênero Candida spp é responsável por cerca de 80% das infecções fúngicas no ambiente hospitalar e constitui causa relevante de infecções sistêmicas em pacientes hospitalizados, especialmente em doentes graves e em imunocomprometidos, com predominância da Candida albicans. A adesão das leveduras às células epiteliais do hospedeiro é um potente estimulador para a formação de hifas, forma invasiva do fungo [1]. Os bioterápicos são medicamentos preparados a partir de produtos biológicos, elaborados conforme a Farmacopeia Homeopática Brasileira (FHB)[2], indicados para tratamento de infecções de etiologia conhecida, empregados com grande sucesso no tratamento clínico destas infecções. Os bioterápicos RC, desenvolvidos pelo médico brasileiro Roberto Costa (RC) são preparados a partir do agente etiológico íntegro e, segundo suas pesquisas, possuem maior capacidade de estimular o sistema imunológico do hospedeiro.

O uso da homeopatia para prevenir sintomas da gripe humana e infecção respiratória aguda: um estudo clínico duplo cego, randomizado, placebo controlado com 600 crianças do Serviço Público de Sáude do Brasil / The use of homeopathy to prevent symptoms of human flu and acute respiratory infections: a double-blind, randomized, placebo-controlled clinical trial with 600 children from Brazilian Public Health Service

In Brazil, homeopathy was implemented in the Public Health Service through the National Policy on Complementary and Integrative Practices of the Health Ministry, published in 2006. Homeopathy appears as a very interesting therapy to be used in the Public Health Services since its medicines are compounded at a very low cost. Considering this interesting scenario to develop research in the Public Health, the Family Health Program (FHP) in Petropolis and the Faculty of Pharmacy at UFRJ started a partnership with the Roberto Costa Institute.(AU)

No Brasil, a homeopatia foi implementada no Serviço Público de Saúde, através da Política Nacional de Práticas Integrativas e Complementares do Ministério da Saúde, publicada em 2006. A homeopatia surge como uma terapia muito interessante para ser usada no serviço público de saúde uma vez que seus medicamentos tem um custo muito baixo. Considerando este cenário interessante para desenvolver uma pesquisa em Saúde Pública, o Programa de Saúde da Família (PSF) de Petrópolis e a Faculdade de Farmácia da UFRJ iniciaram uma parceria com o Instituto Roberto Costa. (AU)

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)

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)

Avaliação da produção de citocinas pelos macrófagos J774. G8 após o tratamento com bioterápicos / Evaluation of cytokine production by J774. G8 macrophages after treatment with biotherapics

Strains of macrophages, such as murine J774.G8 macrophages, are susceptible to influenza A infection [1]. One of the responses to viral infection involves the production of various types of immunostimulatory cytokines by infected cells [2].In all cases, there were no significant differences compared to control groups. However, the production of TNF-? detected in macrophages treated by intact and inactivated biotherapics presented a tendency to increase after infection. In fact, similar results were previously detected in other experiments conducted only with the intact biotherapic [3]. The release of the cytokine MCP1 in all experimental situations presented a tendency to decrease after the viral infection when compared to untreated macrophages. No statistically significant difference was detected in the production of IL 12 and IL 10. These experiments will be repeated to confirm the 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)

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)

The biotherapics Influenzinum and Oscilococcinum in the treatment of influenza virus flu / The biotherapics Influenzinum and Oscilococcinum in the treatment of influenza virus flu

Introduction: The influenza virus flu is a widespread illness which is responsible for hundreds of thousands of deaths annually. About 20% of children and 5% of adults are infected with this virus every year. The disease is highly contagious and its transmission occurs by saliva particles of the infected person, expelled by breathing, talking and coughing [1]. Flu pandemics are generally caused by the appearance of a new subtype of the virus in humans, which occurs as a result of the existing flu in animal species transmitted to humans [2]. Despite the fact there are antiviral drugs, the virus develops mutations, creating resistance to these drugs in few days. Thus, the development of new therapies, including homeopathy, that can prevent and/or treat this disease becomes increasingly necessary. In this scenario, biotherapics appear as drugs that are made from biological products, such as secretions, tissues, organs whose compounding follows the homeopathic pharmacopeia. Objective: This study is a literature review on the treatment of flu with biotherapics used in clinical medicine, namely Influenzinum and Oscilococcinum. Method: Studies on the prescription of biotherapics for the prevention and cure of the flu as well as literature about the history and evolution of Homeopathy were reviewed in the present work. Influenzinum is a biotherapic made from the influenza vaccine from Pasteur Laboratory, while Oscillococcinum is obtained from the lysate of the liver and heart of the goose Anas barbaries. Results: Preliminary results showed that both medicines are widely used in clinical medicine. Influenzinum 9CH is prescribed for flu prevention and treatment, while Oscilococcinum is more used to reduce the severe symptoms in patients who already have the flu. Conclusion: Based on these results, it is possible to say that Influenzinum has a very important role in the prevention and cure of the influenza and Oscilococcinnum is useful in the relief of the symptoms caused by this disease. (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)

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.

The biotherapics Influenzinum and Oscilococcinum in the treatment of influenza virus flu. / The biotherapics Influenzinum and Oscilococcinum in the treatment of influenza virus flu.

Introduction: The influenza virus flu is a widespread illness which is responsible for hundreds of thousands of deaths annually. About 20% of children and 5% of adults are infected with this virus every year. The disease is highly contagious and its transmission occurs by saliva particles of the infected person, expelled by breathing, talking and coughing [1]. Flu pandemics are generally caused by the appearance of a new subtype of the virus in humans, which occurs as a result of the existing flu in animal species transmitted to humans [2]. Despite the fact there are antiviral drugs, the virus develops mutations, creating resistance to these drugs in few days. Thus, the development of new therapies, including homeopathy, that can prevent and/or treat this disease becomes increasingly necessary. In this scenario, biotherapics appear as drugs that are made from biological products, such as secretions, tissues, organs whose compounding follows the homeopathic pharmacopeia. Objective: This study is a literature review on the treatment of flu with biotherapics used in clinical medicine, namely Influenzinum and Oscilococcinum. Method: Studies on the prescription of biotherapics for the prevention and cure of the flu as well as literature about the history and evolution of Homeopathy were reviewed in the present work. Influenzinum is a biotherapic made from the influenza vaccine from Pasteur Laboratory, while Oscillococcinum is obtained from the lysate of the liver and heart of the goose Anas barbaries. Results: Preliminary results showed that both medicines are widely used in clinical medicine. Influenzinum 9CH is prescribed for flu prevention and treatment, while Oscilococcinum is more used to reduce the severe symptoms in patients who already have the flu. Conclusion: Based on these results, it is possible to say that Influenzinum has a very important role in the prevention and cure of the influenza and Oscilococcinnum is useful in the relief of the symptoms caused by this disease.

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.