La solidez de la investigación homeopática fundamental / The strength of fundamental homeopathic research

La investigación fundamental en homeopatía ha avanzado considerablemente en los últimos 20 años: desde estudios exploratorios con animales y plantas hasta la caracterización de los efectos sistémicos de los medicamentos homeopáticos y estudios in vitro con sistemas celulares aislados para evaluar los cambios en los mecanismos de adaptación celular y señalización intracelular frente a tratamientos homeopáticos variables. El número de artículos publicados a lo largo del tiempo ha permitido realizar varias revisiones sistemáticas. Recientemente, la demostración de que los medicamentos homeopáticos podrían modificar las funciones celulares a través de mecanismos epigenéticos (metilación y desmetilación de ADN) preparó el camino para un campo de investigación completamente nuevo. En paralelo, el descubrimiento de las nanopartículas y propiedades físicas específicas de las diluciones homeopáticas ha arrojado luz hacia un campo antes poco conocido, dado que se consideraba que las diluciones homeopáticas no consistían más que de agua. Así las cosas, los retos para el futuro conciernen a la demostración, o no, de la interrelación entre ambos fenómenos.

Fundamental research in homeopathy has much advanced in the past 20 years. From exploratory studies with animals and plants to the characterization of the systemic effects of homeopathic medicines and in vitro studies with isolated cell systems to assess changes in the mechanisms of cell adaptation and intracellular signaling facing variable homeopathic treatments. The amount of articles published over time enabled several systematic reviews. Recently, demonstration that homeopathic medicines might modify cell functions through epigenetic mechanisms (DNA methylation and demethylation) paved the road for a fully new field of research. In parallel, the discovery of nanoparticles and specific physical properties of homeopathic dilutions brought light to a previously poorly known field, as it was believed that homeopathic dilutions consist in nothing but water. Thus being, challenges for the future concern the demonstration, or not, of the interrelationship between both phenomena.

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.

Environmental parameters modulate lead chloride toxicity on Artemia salina

Different environmental conditions can influence the effects of toxic agents on living beings. Recently, a series of experiments performed in Artemia salina submitted to different kinds of intoxication have shown that both, isotherapic and succussed watercan change Artemia salina ́s bio resilience at different levels. Moreover, it seems to vary according to the circalunar cycle. Objective:To verify if circalunar phases and water agitation can modify the toxicity of lead chloride on Artemia salina in vitro. Methodology:Artemia salina cysts were exposed to seawater containing 0.04% of lead chloride (equal to EC10 or 10% effective concentration, previously determined in a pilot study) in 96-well culture plates. Thirty-six experimental repetitions were performed in four series to observe the possible effects of adding stirred water, the so-called succussed water, and the moon phases. The hatched cysts were recorded after 48 hours using a digital microscope (1000x magnification) to identify the hatching percentage and the viability and mobility of the born nauplii. Results:The exposition of cysts to PbCl2 led to an increase in the hatching rate, and it was more evident during the full moon (p = 0.00014) The addition of succussed water into the seawater medium reduced this effect to the baseline levels. An increase in mobility was seen in nauplii born from exposed cysts during the full moon (p = 0.00077), but this effect was not affected by the treatment with succussed water. Discussion:Although the effects of lead chloride EC10 on the increase of nauplii hatching were expected, two environmental variables changed the sensitivity of cysts to this harmful stimulus. The circalunar cycle varied the hatching rate according to the moon phase, even in laboratory conditions, and the addition of succussed water into the medium reduced the hatching rate, even with different intensities according to the moon phase. The organization of nano and microbubbles generated after the succussion of water could be related to this protective effect and can explain, at least partially the effects of high diluted preparations on this biological context. Conclusion:Environmental factors, such as the circalunar cycle and products of water agitation, can modulate the adaptative control of hatching in Artemia salina exposed to lead chloride at EC10.

Characterization of physicochemical markers for homeopathic medicines and biological supernatant samples

Solvatochromic dyes are probes to detect variations on the dipole moment of solvents after the insertion of homeopathic potencies. Recent studies have shown they can be useful tools in laboratory and field studies to detect the activity of homeopathic remedies.Objective: Determine whether solvatochromic dyes can be a diagnostic tool for cells infected by different agents and/or markers to identify the activity of homeopathic medicines. Methods: Ethilicum1cH, Siliceaterra6, 30, 200cH; Zincummetallicum6, 30, 200cH and Phosphorus6, 30 and 200cH were analyzed by pouring the samples (in a 1:60 rate) into a series of seven dyes (rhodamine, ET 33, ET 30, coumarin 7, NN DMIA, Nile red, methylene violet) diluted in absolute ethanol using pre-established working concentrations. Oscillations of dye absorbance were observed at visible light spectrophotometry according to the remedy and potency. Water and succussed water were used as controls. In a second moment, the absorbance profile of the remedies will be compared with those of biological samples (supernatants) and checked with the biological effect previously obtained from each treatment.Supernatants of RAW 264.7 macrophages stimulated by Calmette-Guérin bacilli (BCG) or infected with Encephalitozoon cuniculiwill be analyzed. Results: Preliminary results have shown that Siliceaterra6cH, Phosphorus30 and 200cH and Zincummetallicum6, 30 and 200cH reduced the absorbance of methylene violet (p=0.01). Repetitions and analysis of supernatants are expected to be performed in the next steps of the study. Future perspectives: Establish a pattern of reactivity of the studied medicines with different dyes and the putative relation with the corresponding supernatants, as an attempt to obtain a "physicochemical signature" for each kind of infection and/or treatment.

Carcinosinumproduces phenotypic changes of 4T1 tumor cells in vitro

Considering that there are few published studies that specifically address the exclusive use of Carcinosinumin different potencies and, most of them focused on genotypic and clinical effects, the present study was proposed to identify possible phenotypic changes, including viability, expression of HER-2 and metastatic abilities, using 4T1 cells in vitroas a model. Carcinosinum was tested in different homeopathic potencies (12cH; 30cH; 200cH) mechanically prepared using sterile pure water. The time space between preparing the potencies and using them was 24 hours.The final dilutions were inserted into the culture medium in a volume equal to 10%, at the time of cell seeding. The same succussed vehicle used to prepare the drugs (70% ethanol) diluted 1:100 in sterile pure water was used as control. All treated cells were cultured in 25 mL flasks, with cell density of 5 x 105cells/mL. After 24 hours of treatment, cells were analyzed for apoptosis index using Annexin V kit and the Countess® system. The morphology of 4T1 cells was monitored by staining cell smears with hematoxylin-eosin and Giemsa methods. HER-2 expression was assessed by immunocytochemistry and metalloproteinase activity was assessed by zymography. The determination of the cytokine profile was performed using Cytometric Bead Array (CBA). The samples were evaluated in quadruplicate and the data were analyzed by one-way ANOVA. Carcinosinum30cH presented the highest apoptotic index and reduction of MMP-9-Pro expression; Carcinosinum200cH produced the highest positivity for HER-2 and no specific effect was seen after the treatment with Carcinosinum12cH. No change in cytokine expression was seen among treatments. We conclude that Carcinosinum30cH and 200cH can change phenotypic features important totumor development in vitro. The clinical meaning of these data deserves further investigation.

Effects of homeopathic preparations on raphidiopsis raciborskii (Cyanobacteria) toxicity in artemia salina mode

Cyanobacteria are microorganisms found in different parts of the world. Some genera are cyanotoxins producers a sodium channel blockingneurotoxin (saxitoxins). Some homeopathic preparations have been identified as remedial action on toxicity models in Artemia salina. This study aimed to observe whether homeopathic products influence the toxicity ofR.raciborskiiextract onA.salinaby inducing cyst hatching arrest, anembryo bioresiliencemodel previously developed in our laboratory (Pinto et al., 2021; Mohammad et al., 2022). Thus, previous toxicity testswere carried out on cysts in 96-well plates, using different concentrations of the extract obtained from regular cultivation of R. raciborskii in HCl 0.05M, whose strain, named T3, is kept in the laboratory of Cyanobacteria at FURGS, Brazil.The standardization of toxin concentration was based on an established scale developed at FURGS, in which the number of T3 filaments is associated withspecific saxitoxin concentrationsdefined by chromatography. The concentration of 2.6 µg/L was chosen since it reducedthe cysthatching rate by 30%, the ideal level to observe embryo bioresilience. Then, a screeningstudy with 22 homeopathic preparations was tested blind in three experimental series, in duplicate,against threecontrols (unchallenged, water,and succussed water)for possible toxicity attenuationon Artemia salinacysts hatching rate. Homeopathic medicines were prepared in pure,sterile water from a stock homeopathic solution, one potency below the working potency. After the 1:100 dilution, 100 succussions were made using a robotic arm (Denise, Autic). The medicines were inserted into the seawater on a 10% basis. Due to the high sensitivity of A. salinato the circalunar variations, all experiments were performed during the first quarter moon. Statistical analysis was performed by two-way ANOVA followed by Tukey, with α=0.05. The most significant results indicative of bioresilience improvement were seen after the treatment with Nitric acidum6 cH, Plumbum metallicum6 cH, isotherapic 200 cH, and hydrochloric acid 1 cH being the last one used as a vehicle of the extracts. Thus, these preparations were chosen to be used in further experiments. In conclusion, the Artemia salinamodel has also beenuseful to study bioresilienceimprovement by homeopathic medicines after intoxication with saxitoxin.

Mercury chloride toxicity attenuation of the Brine Shrimp Artemia Salinaafter treatment with Mercurius corrosivusas isotherapic

Isotherapics preparedfromtoxic substances have been described as attenuation factors for heavy metal intoxicationin aquatic animals. Herein, Artemia salinaand mercury chloride were usedas a model to identify treatment-related bioresilience. The aim was to describe the effects of Mercurius corrosivus(MC) in different potencies on Artemia salinacyst hatching and on mercury bioavailability. Artemia salinacysts were exposed to 5.0 µg/mL of mercury chloride during the hatching phase. MC6cH, 30cH, and 200cHwere prepared and poured into artificial seawater. Different controls were used (nonchallenged cysts and challenged cysts treated with water, succussed water, and Ethilicum 1cH). Four series of nine experiments were performed for4 weeks to evaluate the percentage of cyst hatchingconsidering all moon phases. Soluble total mercury (THg) levels and precipitated mercury content were also evaluated. Solvatochromic dyes were used to check for eventual physicochemical markers of MCbiological activity. Two-way analysis of variance (ANOVA) with mixed modelswas used for evaluating the effect of different treatments andthe simultaneous influence of the moon phases on the cystshatching rate, at both observation times (24 and 48 hours).When necessary, outliers were removed, using the Tukeycriterion.Thelevel of significance αwas set at 5%. Significant delay (p<0.0001) in cyst hatching was observed after treatment with MC30cH, compared with the controls. An increase inTHg concentration in seawater (p<0.0018) and of chlorine/oxygen ratio (p<0.0001) in suspended micro-aggregateswas also seen, with possiblerelation with mercury bioavailability. Specific interaction of MC30cH with the solvatochromic dye ET33 (p<0.0017) was found. The other observed potencies of Mercurius corrosivus6 and 200 cH were not significant in relation to the observed groups.The results werepostulated as being protective effects of MC30cH on Artemia salina, by improving its bioresilience.

High-diluted thymulin on Ehrlich tumor growth in mice and the importance of tumor microenvironment

Introduction: The aim of the present study was to describe different biological aspects of Ehrlich tumor in mice, such as body weight evolution, tumor growth rate, histological organization and systemic immune response after treatment with high-diluted thymulin (10-9 M, named 5CH). Methods: Tumor assessment was focused on macro- and microscopic aspects; parameters included occurrence of necrosis, embolism and tumor development, in addition to quantitative analysis of apoptosis (caspase-3), cell proliferation (Ki-67) and angiogenesis (vascular endothelial growth factor - VEGF) by means of specific immunohistochemistry markers. Spleen cell populations were evaluated by flow cytometry analysis. Results: Mice treated with thymulin 5CH exhibited changes in the tumor microenvironment, such as reduced micro-embolism incidence and cytokeratin expression, with increased caspase-3 expression in the tumor cells. These findings indicate some apoptotic activity by the tumor cells induced by the treatment, even though no reduction of the macroscopic tumor mass occurred. No changes in the systemic immune response were detected, as the balance among spleen cell populations remained unchanged. Conclusions: The results indicate that treatment of mice bearing Ehrlich tumor with thymulin 5CH induces some specific changes in the tumor environment. However, it did not influence systemic immunity parameters. Adjuvant use of thymulin 5CH in oncological clinical practice is still a matter of discussion. (AU)

The soundness of homeopathic fundamental research

Fundamental research in homeopathy has much advanced in the past 20 years. From exploratory studies with animals and plants to the characterization of the systemic effects of homeopathic medicines and in vitro studies with isolated cell systems to assess changes in the mechanisms of cell adaptation and intracellular signaling facing variable homeopathic treatments. The amount of articles published over time enabled several systematic reviews. Recently, demonstration that homeopathic medicines might modify cell functions through epigenetic mechanisms (DNA methylation and demethylation) paved the road for a fully new field of research. In parallel, the discovery of nanoparticles and specific physical properties of homeopathic dilutions brought light to a previously poorly known field, as it was believed that homeopathic dilutions consist in nothing but water. Thus being, challenges for the future concern the demonstration, or not, of the interrelationship between both phenomena.

Homeopathy in parasitic diseases

Background: The use of homeopathic medicines has increased, partly because conventional drugs do not always elicit the desired effects, and partly because their side effects might compromise the patient’s’ adherence to treatment. Several studies showed benefits in the use of highly diluted medicines for the treatment of infectious diseases. Aim: The aim of the present review was to perform a critical discussion about aspects of homeopathy and the current status of veterinary experimentation, as well as of the use of highly diluted drugs in infectious and parasitic diseases. The main aspects of effects, therapeutic regimens and / or dynamizations used in various models are discussed. Methods: Articles published since 2000 in journals included in databases PubMed and SciELO and specialized journals sought for and reviewed using keywords “parasitic diseases/homeopathy” and “parasitic diseases/ ultra-dilutions”. Results: Several recent experimental studies demonstrated the biological effect of highly diluted medications on parasitic infections, with reduction of the number of parasites and improvement of the clinical condition of the affected animals. Several articles exhibit problems in the description of methods, which threaten the reproducibility of experiments. Conclusion: The acknowledgment of homeopathy depends on the credibility of investigators. Although research on homeopathy has clearly increased in recent years, relative to both implementation of more consistent methods and description of data and methods, improvement is still required. Precise and detailed descriptions will contribute to advance the use of homeopathy, so that society at large might benefit in actual practice.

Introdução: O uso de homeopáticos tem crescido devido o fato de que medicamentos convencionais algumas vezes não produzem o efeito esperado e também devido os seus efeitos indesejados que comprometem a adesão ao tratamento. Alguns trabalhos mostram os benefícios da utilização de medicamentos ultradiluídos no tratamento de doenças infecciosas. Objetivo: Esta revisão é uma discussão crítica sobre os aspectos da homeopatia, o estado atual da experimentação veterinária e o uso de medicamentos ultradiluídos nas infecções e doenças parasitárias. Os principais aspectos dos efeitos, esquemas de tratamento e/ou dinamizações utilizadas nos diferentes modelos são discutidos. Metodologia: Uma revisão de artigos publicados desde 2000 em revistas indexadas nos bancos de dados PubMed e Scielo e revistas especializadas foi utilizada para pesquisa das palavras-chave: parasitoses/ homeopatia e parasitoses/ ultradiluídos. Resultados: Experimentos recentes demonstraram o efeito biológico dos medicamentos ultradiluídos nas infecções parasitárias, com redução no número de parasitos e melhora da condição clínica nos animais tratados. Alguns artigos apresentaram problemas na descrição da metodologia o que pode comprometer a reprodutibilidade dos experimentos. Conclusão: O conhecimento da homeopatia depende da credibilidade dos grupos de pesquisa. Embora nos últimos anos a pesquisa em homeopatia tenha apresentado um claro desenvolvimento, tanto na implantação de metodologias mais consistentes quanto na descrição dos dados e métodos publicados, muito ainda deve ser melhorado neste campo. Descrições precisas e detalhadas poderiam contribuir para o avanço do uso da homeopatia, o que poderia beneficiar a comunidade em geral , na prática, a partir destes resultados.

A medicina dos druidas e o Viscum album / The medicine of the druids and the Viscum album

Os autores apresentam uma revisão histórica sobre o Viscum album, planta utilizada na Farmacopeia homeopática e sua relação com a cultura Druida.

Biosemiotics and Body Signifier Theory: a way to understand High Dilutions / Biosemiotics and Body Signifier Theory: a way to understand High Dilutions

Since the 80´s Madeleine Bastide and Agnès Lagache have worked on the idea of the Body Signifier Theory, in which the living systems could be defined as ?sensible? systems not only able to self-organize, but also to receive and process non-molecular information according to the Pierce semiotic triad: matrix, receiver and carrier. These ideas were built along 20 years of experimental observations, whose results presented some kind of stereotyped pattern that emerged from cells or animals exposed to high dilutions, according to the similia principle. Curiously, at the same time, classical scientific communities have developed the concept of biosemiotics after the observation that living systems are semiotics entities able to deal with codes and meanings, even in molecule-driven processes. Not only the genetic code, but all regulatory functions in living systems represent coding processes. The development of methodologies to identify and understand these codes and its outputs is the aim of biosemiotic science. Putting both concepts together (body signifiers and biosemiotics), it is possible to recognize strong similarities between their approaches and methodology, allowing logical connections between non-molecular signals and changes in cell interactions patterns. Thus, the general concept of biosemiotic could be a theoretical platform on which the construction of a solid phenomenological description of high dilutions effects could be done.

Inflammation in rats born to mothers treated with dexamethasone 15cH during pregnancy: an immunohistochemical study / Inflammation in rats born to mothers treated with dexamethasone 15cH during pregnancy: an immunohistochemical study

In previous studies, we observed that rats born to mothers treated with dexamethasone 15CH (10-33M) had a higher level of mast cell degranulation and greater arteriolar dilation after the exposure of an inflammatory stimulus, suggesting the possibility of vertical transmission of the effects of ultra-diluted substances between mother and offspring. In this study, a more detailed assessment of the cellular events in acute inflammation was made using techniques of immunohistochemistry. The identification of adhesion molecules expression was made by the markers: anti-CD54 (ICAM-1) and anti-CD18 (?2-Integrin). The identification of inflammatory cells was performed by the markers anti-MAC387 (mononuclear cells) and anti-CD163 (active macrophages). Polymorphonuclear cells were identified by hematoxylin-eosin staining. The number of labeled cells per was recorded, except for the anti-CD54 marker, whose intensity of staining on the endothelial cells was defined by scores assigned by two independent observers. The results point toward to an up regulation of the whole inflammatory process in rats born to mothers treated with dexamethasone 15CH during pregnancy. This conclusion is justified by the following statistically significant (p?0.05) findings: a) bigger mast cell degranulation and increased of arteriolar diameter; b) increased migration of polymorphonuclear cells in relation to the mononuclear cells; c) earlier expression of CD163 in monocytes, d) higher level of adhesion molecules expression.

Triiodothyronine, diluted according homeopathic techniques, modifies the programmed cell death of tadpole tail?s explants / Triiodothyronine, diluted according homeopathic techniques, modifies the programmed cell death of tadpole tail?s explants

High Dilution is a solution beyond the Avogadro limits that, in the dependence of the applied succussion elicits a suppressive or a stimulant effect on a living cell, with a consequent generation of an oscillatory doseeffect curve. According to Bonamin et al. [1], ?Perhaps, the most enigmatic feature regarding the properties of high dilutions is the non-linearity of their effects. In several studies employing in vivo and ex vivo models, especially involving iso-endopathy, an oscillatory potency-effect curve has appeared. The first observations were initially considered as artifacts, but the repetition of this pattern in different studies involving completely different experimental models, in times and places equally different, points out to the existence of a property intrinsic to dynamized systems.? The entire process of anuran amphibian metamorphosis is under thyroid hormones control, included the complete resorption of the tadpole tail. In the present study, we had successfully established a protocol model to culture Rana catesbeiana tadpoles? tail tips in vitro. A random and blind study was performed, with the intent to prove that T3 5.10-24 M (10 cH) modifies the apoptosis induction of T3 100 nM in explants of Rana catesbeiana tadpoles? tail. 60 explants were distributed in three ways: Group A: without T3 action, at pharmacological and HD dose; Group B (test): under the action of T3 100 nM and treated with T3 10 cH (HD); Group C (control): under the action of T3 100 nM and treated with ethanol 70% unsuccussed. After 96 hours of tissue culture, the mean of initial and final area (1.05 vs. 0.98 cm2) and apoptotic index of the explants from Group A were with minimal difference range and for this reason it wasn?t included in the statistical study. In order to identify significant differences in the area and in the apoptotic index of the remainder explants of the 2 groups, B (test) and C (control), we used a student t-test. However, the mean initial and final explants? area from test and control groups were respectively 1.09 vs. 0.22 cm and 1.00 cm vs.0.24 cm, with a mean reduction of 0.87 cm2 and 0.76 cm2, but this difference didn?t achieve statistical significance (p>0.05). In contrast, apoptosis index was significantly higher in test than in control group 11.7 vs. 7.9 (p<0.05), with is confirmed at the table 1.

Reproductive performance of sows inseminated with diluted semen treated with homeopathic medicine

Aims: this study sought to assess the reproductive performance of sows inseminated with sperm treated with homeopathic medicines. Materials and methods: the semen of 2 sexually mature boars age 18 months Pietrain and Duroc cross-bred with similar genetic and reproductive performance were chosen, as well as 125 sows. Sixteen samples of semen were collected and standardized through semen evaluation. Three homeopathic preparations and a placebo (control) were tested on the sperm (n=31/32 per group): Avena sativa 6cH, Pulsatilla nigricans 6cH and Avena sativa 6cH + Pulsatilla nigricans 6cH. Sows were inseminated 3 times with the same estrous diagnostic procedures. Results: there was significant difference (p<0.05) between Avena sativa 6cH and the other 3 groups regarding the return to estrus and parturition rate. There was no significant difference among the groups regarding the number of newborn piglets. Conclusion: these data suggest that homeopathic preparation Avena sativa may be used directly on sperm cells to improve the parturition rate in technified swine farms.

Objetivos: este estudo buscou avaliar o rendimento reprodutivo de suínos inseminados com esperma tratado com medicamentos homeopáticos. Materiais e métodos: o esperma de dois suínos sexualmente maduros de 18 meses de idade híbridos Pietrain e Duroc, com rendimento genético e reprodutivo similares foi selecionado, assim como 125 fêmeas. Dezesseis amostras de esperma foram coletadas e estandarizadas através de avaliação do esperma. Três preparações homeopáticas e um controle (placebo) foram testados no esperma (n=31/32 por grupo): Avena sativa 6cH, Pulsatilla nigricans 6cH and Avena sativa 6cH + Pulsatilla nigricans 6cH. As fêmeas foram inseminadas 3 vezes com os mesmos procedimentos de diagnóstico do estro. Resultados: houve diferença significativa (p< 0,05) entre Avena sativa 6cH e os outros 3 grupos no retorno ao estro e taxa de partos. Não houve diferença significativa entre os grupos no número de recém-nascidos. Conclusão: os dados sugerem que a preparação homeopática Avena sativa pode ser utilizada diretamente nas células espermáticas para melhorar a taxa de partos em fazendas tecnificadas de criação de suínos.

Objetivos; este estudio busco evaluar el rendimiento reproductivo de porcinos inseminados com esperma tratado com medicamentos homeopáticos. Materiales y métodos: el esperma de 2 porcinos sexualmente maduros de 18 meses de edad híbridos Pietrain y Duroc, com rendimiento genético y reproductivo similares fue seleccionado, así como 125 hembras. Dieciséis muestras de esperma fueron recogidas y estandarizadas mediante evaluación del esperma. Tres preparados homeopáticos y un control (placebo) fueron testeados en el esperma (n=31/32 por grupo): Avena sativa 6cH, Pulsatilla nigricans 6cH and Avena sativa 6cH + Pulsatilla nigricans 6cH. Las hembras fueron inseminadas 3 veces con los mismos procedimientos de diagnóstico del estro. Resultados: hubo diferencia significativa (p< 0,05) para Avena sativa 6cH en comparación con los demás grupos en el retorno al estro y tasa de partos. No hubo diferencia significativa entre ninguno de los grupos en el número de recién nacidos. Conclusión: estos datos sugieren que el preparado homeopático Avena sativa puede ser directamente utilizado en las células espermáticas para mejorar la tasa de partos en granjas tecnificadas de porcinos.Palabras llave: porcinos; rendimiento reproductivo; esperma; homeopatía; estudio ciego controlado.

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.

In vitro growth of uropathogenic Escherichia coliisolated from a snow leopard treated withhomeopathic and isopathic remedies: a pilot study / Estudo do crescimento (in vitro) de Escherichia coli uropatogênica isolada a partir de um leopardo das neves e tratada com medicamentos homeopáticos e isopáticos: estudo piloto

This paper reports the results of incubation of a strain of uropathogenic Escherichia coli (UPEC) isolated from a snow leopard - which had died of septicemia secondary to necro-hemorrhagic cystitis - with homeopathic and isopathic remedies. Methods: UPEC was isolated from heart blood and previously typified for virulence factors; it was incubated with homeopathic remedies Cantharis vesicatoria (urinary tract infection affinity), Mercurius solubilis (from symptoms analysis) and nosode prepared from the actual strain, all in dilution 12cH. Results: 2 patterns of bacterial growth were observed, associated to the quality of nutrients in the culture medium; inrich-nutrient medium, nosode of E. coli 12cH had a significant inhibitory effect; in poor-nutrient medium, Merc 12cH exerted significant inhibitory effect. Conclusion: results suggest that the previous conditions of prokaryote systems may influence the in vitro response to homeopathic and isopathic remedies.

Este artigo relata os resultados da incubação de uma linhagem de Escherichia coli uropatogênica (UPEC) isolada a partir de um leopardo das neves, que morreu de septicemia secundária a cistite necrótica-hemorrágica. A UPEC foi tratada com preparados homeopáticos e isopáticos. Métodos: UPEC foi isolada de sangue cardíaco e previamente tipificada para fatores de virulência; foi incubada com o medicamento homeopático Cantharis vesicatoria (afinidade com infecção do trato urinário), Mercurius solubilis (a partir da análise de sintomas) e nosódio preparado a partir da mesma linhagem de bactérias, todas em 12 cH. Resultados: 2 padrões de crescimento bacteriano foram observados, associados à qualidade dos nutrientes do meio de cultura; em meios ricos em nutrientes, nosódio de E. coli 12 cH teve um significativo efeito inibitório; em meio pobre de nutrientes, Merc 12 cH exerceu efeito inibitório significativo. Conclusão: os resultados sugerem que as condições prévias do sistema procarioto estudado podem influenciar as respostas proliferativas in vitro para preparados homeopáticos e isopáticos.

High Dilution of Dexamethasone in gestation and fetal development of a mice

Background: Recently, the use of homeopathy in veterinary medicine has grown significantly, mainly for farm animal practice, because of its usefulness in organic production and low cost. There is a wide range of veterinary products available in the marketoften used in females. However, the effect of these products in the litter and derived products for human consummation is completely unknown. Aims: this study sought to develop an experimental model to study the putative effects of high diluted substances in newborns after chronic exposure of females. Methods: based on previous studies, the chosen test substance was dexamethasone 15cH; adult female Balb/c mice were divided into 4 groups: a) treated with PBS (control); b) treated with dexamethasone 15 cH; c) treated with dexamethasone 15cH + dexamethasone 4 mg/kg and d) treated with dexamethasone 4 mg/kg. All medicines were administered subcutaneously, 3 times a week, in females from the first day of pregnancy up to the 20th day after parturition (end of lactation period). TDevelopment of the offspring was evaluated daily for 15 days after birth. Parameters evaluated were: female and offspring viability, number of newborns, time for eye opening, pinna opening, fur growth and postural reflex. Results: the group treated with dexamethasone 15cH showed 39% increase in mortality rate 39 days after the beginning of treatment and 35% increase in fetal mortality at the end of gestation (p=0.0049). Females treated with dexamethasone 4mg/kg + dexamethasone 15cH showed 100% of fetal mortality. After parturition newborn survival in animals exposed to dexamethasone 4 mg/kg was higher than the control (p=0.0002). All other parameters of neonatal development were unchanged among groups. Conclusions: these data point to adverse effect when using high diluted dexamethasone during gestation detectable by this experimental model in Balb/c mice.

Behavior of rats treated with Rhus toxicodendron 200cH / Conducta da ratones tratados con Rhus toxicodendron 200cH

One of the main pathogenetic characteristic of Rhus toxicodendron (Rhus-t) is the presence ofarticular pain and aggravation on standing, which improve only by motion. The present studyproposes an experimental model to evaluate the action of Rhus-t 200cH. Rats were divided into 3groups according to treatment received (Rhus-t, diazepam and water); each group was furtherdivided into two sub-groups according to the initial pattern of behavior (hyperactive andhypoactive) as assessed by open-field procedure. A second evaluation of behavior performed 24hours later pointed out to the effects of the medications under study. Results were analyzedthrough Kruskal-Wallis/Dunn’s test, with a level of significance p 0,05.

Uma das características patogenéticas principais de Rhus toxicodendron (Rhus-t) é a presença de dor articular e agravação em pé, melhorando exclusivamente com o movimento. O presente estudo propõe um modelo experimental para avaliar a ação de Rhus-t 200cH. Ratos foram divididos em 3 grupos de tratamento (Rhus-t, diazepam e água); cada grupo doi subdividido em 2 subgrupos de acordo com o padrão inicial de conduta (hiper e hipoativos) em procedimento de campo aberto. Uma segunda avaliação da conduta, realizada 24 horas após, indicou o efeito dos medicamentos sob estudo. Os resultados foram analisados pelo teste de Kruskal-Ealis/Dunn com nível de significância p=0,05.

Estudo crítico de neoplasias cutâneas em cães / Critical study of cutaneous neoplasias in dogs

Introdução - Neoplasias tegumentares são comuns no cão doméstico, representando aproximadamente 30 a 40% de todas as afecções tumorais nesta espécie. Este estudo objetivou detectar as neoplasias cutâneas mais freqüentes em cães, bem como seu comportamento biológico, correlacionando-as à raça, sexo e idade, com intuito da caracterização dos padrões de ocorrência desta enfermidade numa amostragem de pacientes, segundo os critérios de identificação mencionados. Material e Métodos - Cem cães acometidos por tumores cutâneos solitários foram atendidos no serviço de cirurgia do Hospital Veterinário da Universidade Paulista e caracterizados segundo padrão racial, sexo e faixa etária. Todos animais foram fisicamente avaliados e suas respectivas lesões submetidas à análise histopatológica. Resultados - Dentre os cães estudados, 64 (64%) eram de raça definida e destes, os mais acometidos foram Poodle (n = 14) (23%), Pastor Alemão (n = 13) (21%) e Boxer (n = 8) (13%), respectivamente. Dos 100 animais avaliados, 57 (57%) eram fêmeas e 43 (43%) machos; 46 (46%) apresentavam idade entre seis e dez anos, 29 (29%) igual ou superior a 11 anos e 25 (25%) até cinco anos. O estudo micromorfológico detectou 81 neoplasias malignas (81%) e 19 benignas (19%), das quais 42 (42%) eram mesenquimais, 35 (35%) epiteliais, 15 (15%) de células redondas e 8 (8%) melanocíticas. Conclusão - Por meio dos resultados obtidos, esta pesquisa concluiu que cães das raças Poodle, Pastor Alemão e Boxer, fêmeas, com idade entre seis e dez anos foram os mais acometidos por neoplasias cutâneas. Constatou-se também, que a maior parte dos tumores tegumentares eram malignos e de origem mesenquimal.

Introduction - Integumentary neoplasias are common in domestic dogs, representing approximately 30 to 40% of all the tumoral affections in this specie. This study aimed at detecting the most frequent cutaneous neoplasias in dogs, as well as their biological behavior, correlating them to breed, sex and age, intending at characterizing the occurrence patterns concerning this illness in a sampling of patients, according to aforementioned identification criteria. Material and Methods - One hundred dogs taken ill by solitary cutaneous tumors, was attended on the surgery service of the Veterinary Hospital of Universidade Paulista and characterized pursuant breed, sex and age. All animals were physically evaluated and their respective lesions undergone to the histopathological analysis. Results - Among the studied dogs, 64 (64%) were purebred and from those, the most taken ill were Poodle (n = 14) (23%), German Shepherd Dog (n = 13) (21%) and Boxer (n = 8) (13%), respectively. Within 100 evaluated animals, 57 (57%) were female and 43 (43%) males; 46 (46%) presented age between six and ten years, 29 (29%) equal or above 11years and 25 (25%) up to five years. The micro-morphological study detected 81 malignant neoplasias (81%) and 19 benignant (19%), of which 42 (42%) were mesenquimal, 35 (35%) epithelial, 15 (15%) of round cells and 8 (8%) melanocytic. Conclusion - Through the obtained results, this research concluded that Poodle, German Shepherd Dog and Boxer breeds, females, with age between six and ten years were most taken ill by cutaneous neoplasias. It was noticed also that most of integumentary tumors were malignant and had mesenquimal origin.