NMR analysis: potential and challenges as a homeopathic basic research tool

Homeopathic ultra-diluted solutions surpass the threshold of matter dispersion and, consequently, their chemical constitution is similar to inert solvent. Nevertheless, randomized clinical trials have shown that the clinical effects of these homeopathic medicines are superior to placebo1. Nuclear magnetic resonance (NMR) is one of the most promising techniques to detect physicochemical alterations induced by homeopathic procedures2,3. Aims: To investigate T2 NMR relaxation times of Zincum metallicumand lactose dynamized samples. Methodology: Zincum metallicumsamples were ground until 6dH using lactose as an excipient. Subsequently, these samples were dynamized with ultrapure water to produce 8dH, 9dH, 10dH,and 11dH. Lactose dynamized samples (6dH-11dH) were used as control. Aliquots of 540µl of each sample were diluted with 60µl of deuterated water (D2O) in 5mm tubes. The analyses were carried out in Bruker Ascend TM 500MHZ spectrometer at 288 K. Results and discussion: The Zincum metallicumand lactose T2 relaxation times were very similar, except for Zincum metallicum8dH, which presented a value of 1.226 in comparison to 1.036 of lactose 8dH. The following T2 values were registered: 1.287 -9dH; 1.413 -10dH; 1.467 -11dH, and 1.303 -9dH; 1.400 ­10dH; 1.350 -11dH, for Zincum metallicumand lactose, respectively. The differences detected in 8dH samples are probably due to the presence of lactose in the first dilution step, in which 1 part of the 6dH triturated mixture was diluted in 9 parts of water, to prepare 7dH. Following this homeopathic procedure, 8dH solutions remain around 1% of lactose which could be influenced by the T2 values registered.Conclusion: These preliminary results showed the possibility to apply the NMR technique to evaluate the influence of dynamization in the relaxation parameters. Further studies should be carried out with other potencies and/or other homeopathic substances, in addition to the evaluation of T1 and the T1/T2 parameters, as previously described by other groups.

Viscumalbum L. homeopathic mother tinctures: metabolome and antitumor activity

Viscum album L. is a semi-parasitic plant with antitumor activity attributed to theaqueous extracts. However, European V. album ethanolic extracts (VAE) have also demonstrated invitro activity in tumor models. Aims: Evaluate the metabolic profiles of fifty VAE harvested duringsummer and winter seasons and their antitumor activity through 2D and 3D models. Methodology:VAEwerepreparedbymacerationfrom:V.albumsubsp.albumgrowingonMalus domestica,Quercus sp.and Ulmus sp.; V. album subsp. austriacum from Pinus sylvestris; V. album subsp. abietis from Abies alba.Chemical analyses were performed through liquid chromatography coupled with high resolutionmass spectrometry and Partial Least Squares Discriminant Analysis (PLS-DA) was performed in theMetaboanalyst 4.0. The antitumor potential of the selected VAE was evaluated in 2D and 3D models(MDA-MB-231 cancer cells) by MTT, crystal violet and glycolytic pathway analysis. Results anddiscussion:Thefirst3principalcomponentsinPLS-DAexplained60%and40%ofdatavariationin positive andnegativemodesrespectively.Threegroupswereformedandshowedchemicalsimilarityamong V. album subspecies. The compounds responsible for group separation were tentativelyidentifiedas:pinobankasinornaringenin hexoside;isorhamnetin-3-hexoside,meglutolanddifferent aminoacids.ThesummerVAEat0.5%v/vinducedhighercytotoxicdamagethanthewinterpreparations, and Abies alba and Quercus sp. VAE promoted 49% and 42% reduction of tumorviability in 3D model (72h incubation), respectively. MDA-MB-231 glycolytic pathway in 2D modelshowed a decrease in the glucose consumption and extracellular lactate production. Also, PFK (6-phosphofructo-1-kinase)andPK(Pyruvatekinase)activitieswereinhibitedbyAbiesalbaandQuercus sp. VAE at 48h of incubation. Conclusion: VAE extracts showed different metabolomes andthe glycolytic pathway should be an important target involved in the inhibition of tumor growth bytheseextracts

In vitro evaluation of the effects of Viscum album homeopathic preparations on murine melanoma cells

Viscum album extract (VA) is a complementary treatment in cancer, with in vitro and in vivo cytotoxic effects on several tumor types when applied in phytochemical doses. However, highly diluted ethanolic homeopathic preparations' effects and mechanisms need further study. Aims:To assess the in vitro effects of highly diluted VA from the subspecies V. album abietis and V. album album at different potency levels in different dilution ratios on murine melanoma cells. Methodology:The VA mother tinctures (MT)from Abies alba (MTA) and Quercus robur (MTQ) were prepared with summer and winter samples, harvested in Switzerland. They were submitted to homeopathic ethanolic maceration and a subsequent dynamization process. MTA, MTQ and the following respective potencies were tested in B16F10 murine cells: 3x, 12x, 30x, 6cH, 12cH, 200cH, 2LM, 3LM, and 5LM. Dynamized water, dynamized and non-dynamized ethanol, and carboplatin were used as control groups. The mitochondrial activity and cell viability analysis were performed at 1, 24, 48, and 72 hours by in vitro incubation. MTA and MTQ harvested in summer, as well as 12x, 200cH and 5LM potencies were also tested to cell apoptosis and necrosis markers, reactive oxygens species (ROS) production, inflammatory cytokines profile, cell morphology, and migratory capacity. Results and discussion: MTA and MTQ induced a decrease in cell metabolism and higher cytotoxicity within 1 hour, with significant morphological changes and increased production of ROS and inflammatory cytokines. Both homeopathic dilutions 12x and 5LM showed an influence on cell metabolism, cell replication, and oxidative stress modulation with inflammatory cytokines, mitosis, and migration pattern changes. On the other hand, Quercus robur and Abies alba 200cH showed increased on cytotoxicity and ROS levels, respectively. Conclusion:The in vitro effects of Viscum album homeopathic solutions in melanoma cells highlight the promising antitumoral potential and reinforce the need for further research to better understanding their mechanisms of action.

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

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

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.

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.

Putative protective effect of Cadmium chloride high diluted solution on LLC-PK1 cell intoxicated by high concentration of this same metal: an isopathic in vitro assay

Cadmium is an important toxic environmental heavy metal. Several studies have demonstrated that a major site of cadmium toxicity in humans and in other animals is the proximal tubule of the kidney. A well established model for nefrotoxicity is the use of in vitro technique with proximal tubule epithelial cell lines, as LLC-PK1. Herein, we have the intention to study the possible protective effect of high diluted CdCl2 solutions. In a blinding way, LLC-PK1 cells were pre-treated with high diluted cadmium chloride in the potencies 10 cH, 15 cH and 20cH. After 4 days, these cells have received CdCl2 in a pre-determined toxic concentration. The cell viability was assessed by MTT assay. We have identified a protective effect of two CdCl2 high diluted solutions, 10 cH and 20 cH, when cells were intoxicated by sublethal CdCl2 concentration. The results indicate that probably the high dilutions have an expressive action on cells in sublethal intoxication.

O Cádmio é um contaminante ambiental relevante. Muitos estudos demonstram que o sítio de toxicidade em humanos e outros animais é o túbulo proximal do rim. Um modelo bem estabelecido para nefrotoxicidade é o uso de técnicas in vitro com linhagens de células epiteliais do túbulo proximal, conhecidas por LLC-PK1. Assim, nossa proposta foi a de estudar os eventuais efeitos protetores de uma alta diluição de CdCl2. Em um ensaio cego, células LLC_PK1 foram pré-tratadas com altas diluições de cloreto de cádmio nas diluições 10 cH, 15 cH e 20 cH. Após 4 dias, estas células receberam CdCl2 em uma concentração tóxica, previamente deteminada. A viabilidade cellular foi estudada por ensaios MTT. Observamos um efeito protetor para duas altas diluições de CdCl2, 10 cH e 20 cH, quando as células foram intoxicadas por concentrações subletais de CdCl2. Estes resultados indicam a possibilidade de que altas diluições tenham ação expressiva em células, em intoxicações subletais.

El Cádmio es un metal pesado com relevante acción tóxica en el medio ambiente. Varios estudios han demostrado que un sitio importante de la toxicidad del cadmio en los humanos y en otros animales es el túbulo proximal del riñón. Un modelo bien establecido de nefrotoxicidad es el uso de la técnica in vitro con células epiteliales del túbulo proximal, como las LLC-PK1. Estudiamos el posible efecto protector de soluciones altamente diluidas de CdCl2. Com uma metodologia em ciego, las células LLC-PK1 fueron pre-tratados con cloruro de cadmio altamente diluídos en las potencias 10 cH, 15 cH y 20 cH. Después de 4 días, estas células han recibido CdCl2 en una concentración tóxica predeterminado. La viabilidad celular se evaluó por el ensayo MTT. Hemos identificado un efecto protector de dos soluciones de altamente diluída de CdCl2, 10 cH y 20 cH, cuando las células se intoxicaron por concentración CdCl2 subletales. Los resultados indican que probablemente las altas diluciones tienen una acción expresiva en las células, en la intoxicación subletal.

Physical chemical and citotoxic evaluation of highly diluted solutions of Euphorbia tirucalli L. prepared through the fifty milesimal homeopathic method / Avaliação físico-química e citotóxica de soluções altamente diluídas de euphorbia tirucalli l. Preparada pelo método cinqüenta milesimal (lm) / Evaluación físico-química y citotóxica de soluciones altamente diluídas de Euphorbia tirucalli L. preparadas por el método homeopático cincuenta milesimal (LM)

Motivação: Embora Hahnemann tenha descrito o método LM na 6ª. Edição do Organon da Arte de Curar, poucas pesquisas tem sido feitas sobre as propriedades físico-químicas destas preparações. Além disso, ainda não existem evidências que apontem para a correlação entre propriedades físico-químicas e os efeitos biológicos das altas diluições. Objetivos: determinar características físico-químicas das preparações LM tais como condutividade elétrica, pH e índice de refração, bem como o efeito biológico em modelos experimentais. Materiais e Métodos: foi analisada uma série de diluições desde 1 lm até 10 lm de Euphorbia tirucalli L., preparada a partir do látex e do suco da planta. Para considerar as características sazonais desta planta, usamos 2 populações diferentes, colhidas uma em Junho de 2007 e outra em Maio de 2008. Além disso, o efeito citotóxico da Euphorbia tirucalli L. 5 lm foi testado em células humanas cancerosas (MCF7) através de ensaio MTT. Conclusões: Algumas diferenças entre as duas populações foram observadas. No entanto, nenhuma correlação clara pode ser observada entre as propriedades físico-químicas e a atividade biológica.

Background: although Hahnemann described the fifty-milesimal (LM) method in the 6th edition of the Organon of the Medical Art, very little research has been carried out on the physical chemical properties of these homeopathic preparations. Furthermore, there is still no evidence allowing for the correlation between the alleged physical chemical properties and the biological effects of high dilutions. Aims: to evaluate physical chemical characteristics of LM preparations including electrical conductivity, pH and refraction index, and their effect on biological experimental models. Materials and methods: preparations tested for physical chemical analysis were dilutions 1 lm to 10 lm of Euphorbia tirucalli L. prepared from the latex and the juice of the plant. To rule the seasonal characteristics of this plant, 2 different populations were used, one collected in June 2007 and the other in May 2008. Furthermore, the cytotoxic effect of Euphorbia tirucalli 5 lm was tested on human breast cancer cells (MCF7) through MTT assay. Conclusions: Some differences among the two collections were observed. However, any clear correlation could be observed between physical chemical properties and biological activity.

Motivación: Aunque Hahnemann describió el métodocincuenta milesimal (LM) en la 6 ª edición del Organon del Arte Médica, muy poca investigación se ha realizado sobre las propiedades físico-químicas de estos preparados homeopáticos. Por otra parte, todavía no hay pruebas que permitan la correlación entre las presuntas propiedades físico-químicas y los efectos biológicos, de las altas diluciones. Objetivos: evaluar las características físico-químicas de los preparados LM, incluyendo conductividad eléctrica, pH y el índice de refracción, y su efecto en modelos biológicos experimentales. Materiales y métodos: las muestras para el análisis físico-químico fueron diluciones 1 a 10 lm de Euphorbia tirucalli L. preparadas a partir del látex y el jugo de la planta. Para descartar las características sazonales de esta planta, dos poblaciones diferentes fueron utilizadas, una recogida en junio de 2007 y otro en mayo de 2008. Además, el efecto citotóxico de Euphorbia tirucalli 5 lm fue probado en células humanas de cáncer de mama (MCF7) a través del ensayo MTT. Conclusiones: Algunas diferencias entre las dos colecciones se observaron. Sin embargo, ninguna correlación clara se observó entre las propiedades físicas- químicas y actividad biológica.

A homeopatia e os modelos experimentais para a compreensão das propriedades físico-químicas e biológicas dos sistemas dinamizados / Homeopathy and experimental models for the undestanding of the physical-chemical and biological properties of dynamized systems

Sistemas dinamizados (SD) têm sua origem através da interação entre dois processos: diluição e agitação. Alguns autores têm sugerido que o processo de dinamização é capaz de alterar as propriedades físicas e químicas da água. Entretanto nos dias de hoje, pouco se conhece acerca dos SD, em parte porque a própria dinâmica da água, assim como suas propriedades físico-químicas não são completamente compreendidas. O presente artigo revisa o estado atual dos conhecimentos a esse respeito e sumariza a linha de pesquisa sendo desenvolvida no Brasil, com pouco suporte em termos de recursos, pelo grupo liderado pela autora.

Crude ethanol extract from babassu (Orbignya speciosa): cytotoxicity on tumoral and non-tumoral cell lines

Plant-derived substances have been considered as important sources of drugs, including antineoplasic agents. Babassu mesocarp is popularly used in Brazil as a food additive, and in popular medicine against several conditions, such as inflammations, menstrual pains and leukaemia. From babassu Orbignya speciosa (Mart.) Barb. Rodr. [Arecaceae (Palmae)] epicarp/mesocarp, an ethanol extract was prepared and named OSEME, which was tested on the viability,morphology and metabolism of several cell lines, such as the leukaemic cell lines, HL-60, K562 and the latter multidrug resistant counterpart K562-Lucena 1, the human breast cancer cell line MCF-7, the mouse fibroblast cell line 3T3-L1 and fresh human lymphocytes. OSEME promoted a dose-dependent decrease on the viability of all cells. This effect was much more pronounced on the tumoral cell lines than on non-tumoral cells, a phenomenon revealed by the dose of OSEME which promotes half of maximal effect (ID50). The decrease on viability was followed by shrinkage of cells, alteration on their morphology, and a markedly nuclear condensation. Curiously, stimulation of 6-phosphofructokinase activity (6.6-times) was observed on HL-60 cells, treated with OSEME, when compared to control treated with ethanol (vehicle). These results support evidences to suggest OSEME as a promising source of novel antineoplasic agents.

Substâncias derivadas de plantas têm sido usadas como importante fonte de agentes antineoplásicos. O mesocarpo do babaçu é popularmente usado no Brasil como suplemento alimentar e na medicina popular para o tratamento de várias afecções, tais como: inflamações, cólicas menstruais e leucemia. A partir do epicarpo/mesocarpo do babaçu Orbignya speciosa (Mart.) Barb. Rodr. [Arecaceae (Palmae)] foi preparado um extrato etanólico, denominado OSEME, o qual foi incubado com as seguintes linhagens humanas leucêmicas: HL-60, K562 e a sua derivada resistente a múltiplas drogas, K562-Lucena 1; além destas, foram testadas a linhagem humana de câncer de mama, MCF-7; a linhagem de fibroblastos de camundongo, 3T3-L1 e linfócitos humanos de sangue periférico. OSEME promoveu diminuição da viabilidade em todas as linhagens celulares testadas de maneira dose-dependente. Este efeito foi mais pronunciado sobre as linhagens celulares tumorais quando comparado às não tumorais, o que foi revelado pela dose de OSEME capaz de promover metade do efeito máximo (ID50). A diminuição da viabilidade foi acompanhada por danos sobre a morfologia celular com pronunciada condensação citoplasmática e nuclear. Curiosamente, quando a linhagem HL-60 foi tratada com OSEME, foi detectado um aumento de 6,6 vezes da atividade da enzima 6-fosfofrutoquinase, quando comparado ao grupo controle (células tratadas com o veículo etanol). Esses resultados sugerem que OSEME pode ser uma promissora fonte de novos agentes antineoplásicos.

Chronic toxicological effects of high diluted solutions of Aveloz (Euphorbia tirucalli L. ) on healthy mice: a preliminary study / Efeitos toxicológicos crônicos de altas diluições de Aveloz (Euphorbia tirucalli L. ) em camundongos sadios: estudo preliminar

The latex extracted from Euphorbia tirucalli, a plant popularly known as Aveloz, is used in complementary medicine to induce tumor regression. However, as this latex has toxic effects when administered orally in ponderal doses, the present study was designed to assess the effects of high dilutions in healthy mice over a period of 18 weeks. The Aveloz latex-high diluted solutions (latex-HD) were obtained through the interaction of two processes: 1:100 dilution in mass and succussion, using ethanol 70% as a solvent, in the homeopathic dilutions of 5, 15 and 30cH, following Farmacopéia Homeopática Brasileira. Control solutions without latex were compounded (ethanolic-HD) in the same dilutions and were administered simultaneously. The animals which received latex-HD 30cH showed a significant increase in food consumption (p < 0.05) without significant difference in weight gain. In regards to water consumption, no statistical difference was shown when different Aveloz latex-HD groups were compared, apart from the group that received 1 drop of pure latex in water, which presented a significant increase (p < 0.05) in this parameter.

O látex extraído de Euphorbia tirucalli, planta conhecida popularmente como Aveloz, é utilizado em medicina complementar para induzir regressão tumoral. No entanto, como este látex tem efeitos tóxicos quando administrado por via oral em doses ponderais, este estudo foi desenhado para avaliar os efeitos de altas diluições do mesmo em camundongos sadios por um período de 18 semanas. As altas diluições do látex do Aveloz (látex-HD) foram obtidas através da interação de dois processos: diluição 1:100 em massa e sucussão, utilizando etanol 70% como solvente, nas diluições homeopáticas 5, 15 e 30cH, de acordo com a Famracopéia Homeopática Brasileira. Soluções de controle sem o látex (etanol-HD) foram compostas nas mesmas diluições e ministradas simultaneamente. Os animais que receberam látex-HD 30cH mostraram aumento significativo no consumo de alimento (p < 0,05) sem diferença significativa em ganho de peso. A respeito do consumo de água, não foram observadas diferenças significativas entre os diferentes grupos látex-HD, enquanto que o grupo que recebeu 1 gota de látex puro em água apresentou aumento significativo neste parâmetro (p < 0,05).

High dilutions of Euphorbia tirucalli L. (aveloz) modify the viability and glycolytic metabolism of cell lines / Altas diluições de Euphorbia tirucalli L. (aveloz) modificam a viabilidade e o metabolismo glicolítico de linhas celulares

The latex of Euphorbia tirucalli L. (Aveloz) is popularly used to treat cancers from diverse sources such as: breast, prostate, lung and kidney. Furthermore, high dilutions of this latex (latex-HDs) have been successfully used in the treatment of tumors, although the mechanisms involved in this antitumoral activity are not yet known [...] Latex-HDs were obtained through the interaction of two procedures: 1:100 dilution in mass and agitation, using 2 solvents, water and 70% ethanol in homeopathic preparations 5, 15 and 30cH, following the Brazilian Homeopathic Pharmacopeia. Cell lines were incubated with 1 % and 10 % of latex-HDs for 24 hours. Controls treated with similar preparations without latex were incubated under the same conditions. Cell viability was analyzed by MTT assay and morphological features were assessed by May-Grunwald-Giemsa method. Cell ultrastructure was analyzed by transmission electron microscopy. Metabolic alterations were detected by spectrophotometric assay for the enzyme 6-phosphofructo-1-kinase (PFK-1) activity. 1% of ethanolic latex-HDs induced no alterations in MelanA cells viability, however Aveloz 15cH induced an increase in MCF7 cells viability. After aqueous treatment, MelanA cells viability decreased in almost all systems, whereas in MCF7 cells, an increase was detected in dynamized water systems and Aveloz 15CH. However, only water 15cH increased cell viability in comparison to control and non-dynamized water. No alterations in MCF7 morphology and ultrastructure were visualized through light microscopy or electronic microscopy, respectively. Interesting results were detected when we studied the glycolytic metabolism of both cells, giving support to evidences showing that HDs interfere in the metabolism of cell lines. Further studies are needed to better understand the mechanisms involved in this in vitro biological response to HDs.

O látex de Ephorbia tirucalli L. (aveloz) é usado popularmente no tratamento de diversos tipos de câncer incluindo mama, próstata, pulmão e rim. Além do mais, altas diluições desse látex (latex-HDs) têm sido utilizadas com sucesso no tratamento de tumores, embora o mecanismo dessa ação ainda seja desconhecido [...] As látex-HDs foram obtidas através de 2 procedimentos: diluição 1:100 em massa e agitação, com 2 solventes, água e etanol 70%, nas diluições homeopáticas 5, 15cH e 30cH de acordo com a Farmacopéia Homeopática Brasileira. As linhas celulares foram incubadas com látex-HDs 1% e 10% por 24 horas. Foram preparados controles similares sem o látex e incubados sob as mesmas condições. A viabilidade celular foi analisada através do método May-grunwald-Giemsa. A ultrastrutura celular foi analisada através de microscopia eletrônica de transmissão. As alterações metabólicas foram detectadas por ensaio de espectrofotometria para a atividade da enzima 6-fosfofructo-1-quinase (PFK-1). Látex-Hds em etanol 1% não induziram qualquer alteração na viabilidade das células MelanA, mas Aveloz 15cH induziu aumento da viabilidade das células MCF7. Após tratamento, a viabilidade das células MelanA diminuiu em quase todos os sistemas, enquanto que nas células MCF7 foi detectado aumento nos sistemas aquosos e Aveloz 15cH, mas só água 15cH aumentou a viabilidade celular por comparação ao controle e água não dinamizada. Não foram detectadas alterações na morfologia celular por microscopis de luz quando se estudou o metabolismo glicolítico em ambas as células, demonstrando que os padrões de resposta celular dependem do tipo de solvente, metabolismo celular e procedimento de diluição. Esta é a primeira vez que se mostra que as altas diluições interferem no metabolismo de linhas celulares. Outros estudos são necessários para se compreender melhor os mecanismos envolvidos nesta resposta in vitro a altas diluições de aveloz.

Modeling physical-chemical properties of high dilutions: an electrical conductivity study / Modelagem de propriedades físico-químicas das altas diluições: o estudo da condutividade elétrica

The most common way to perform succussions is to place a liquid preparation inside a glass vessel and beat it vigorously against a hard elastic surface, either manually or using a mechanical apparatus. This procedure has been assumed able to transfer mechanical energy to the molecular level, where it becomes available to perform chemical work. Such interpretation has been enforced by observed changes in the electrical conductivity (EC) of High Dilutions (HD) due to succussion. In order to address this question, we compared the electrical conductivity changes of HD prepared from Vincristine sulfate (VCR) samples with those of an inert solvent. Samples were produced through manual and mechanical succusions in order to observe the influence of bubbles production. The results confirmed the timing of EC changes but these were equivalent for VCR and solvent, except for VCR 1cH samples. Also, the production of bubbles does not affect the EC in an extent able to distinguish succussion procedures. We concluded that the physical-chemical properties of HD can be modeled by chemical and diffusive mechanisms typical of distilled water.

A maneira mais comum de realizar sucussões é manter uma preparação líquida dentro de um frasco de vidro, e agitá-la violentamente usando um dispositivo mecânico ou batendo-a, manualmente, contra um anteparo rígido, porém elástico. Este procedimento tem sido interpretado como um mecanismo capaz de transferir a energia mecânica da agitação, para níveis moleculares, onde ela se tornaria disponível para a realização de trabalho químico. Tal interpretação tem sido estimulada pela observação de alterações na condutividade elétrica (CE) de Altas Diluições (HD), devido à sucussão. Visando modelar esta questão, comparou-se a alteração da CE de amostras preparadas a partir de Sulfato de Vincristina (VCR) com amostras equivalentes (controle) preparadas apenas com o solvente água destilada. As amostras foram produzidas por sucussão mecânica e manual, a fimd e observar a influência da produção de bolhas. Os resultados confirma que a CE varia com o tempo, porém tais variações são equivalentes para VCR e controle, exceto para as amostras VCR 1cH. Também, a produção de bolhas não afeta a CE de forma a permitir usá-la para distinguir os diferentes procedimentos de sucussão. Concluimos que as propriedades físico-químicas da HDs podem ser modeladas por mecanismos químicos e difusivos, típicos para a água destilada.