Effects of Homeopathic Phosphorus on Encephalitozoon cuniculi-Infected Macrophages In-Vitro.

INTRODUCTION: Encephalitozoon cuniculi (E. cuniculi), a fungus that acts as an intracellular pathogen, causes a marked neurological syndrome in many host species and is a zoonotic concern. Although no well-established treatment for this syndrome is known, previous successful clinical experience using homeopathic phosphorus has been described in which symptom remission with no mortality occurred in 40/42 animals by means of unknown immunological mechanisms. The latter observation was the main motivation for this study. OBJECTIVE: To verify, in an in-vitro model, if macrophages infected with E. cuniculi can change in function after treatment with different potencies of phosphorus. MATERIALS AND METHODS: RAW 264.7 macrophages were infected with E. cuniculi in-vitro and treated with various homeopathic potencies of phosphorus. The vehicle was used as a control solution (0.06% succussed ethanol). After 1 and 24 hours, the following parameters were analyzed: parasite internalization (by the Calcofluor staining method), lysosome activity (by the acridine orange method), cytokine/chemokine production (by the MAGPIX system), and cell ultrastructure. Automatic image analysis was used when applicable, and the experiments were performed in triplicate. RESULTS: Treatment with vehicle alone increased interleukin (IL)-6, tumor necrosis factor alpha and monocyte chemotactic protein -1 production (p ≤ 0.05) and reduced the number of internalized parasites (p ≤ 0.001). A progressive and time-dependent increase in RANTES (regulated on activation, normal T-cell expressed and secreted) and lysosome activity (p ≤ 0.002) was observed only after treatment with the highest potency of phosphorus (Phos 200cH), together with decreased apoptosis rate, intense parasite digestion, and the presence of non-internalized spores. CONCLUSIONS: Phos 200 cH has a modulatory action on the activity of infected macrophages, especially a specific increase in RANTES, a key element in the prognosis of E. cuniculi-infected and of immunosuppressed patients bearing infections.

Biological Actions, Electrical Conductance and Silicon-Containing Microparticles of Arsenicum Album Prepared in Plastic and Glass Vials.

INTRODUCTION: According to the "silica hypothesis" formulated to explain homeopathy, the information of starting materials would be transferred to cells by silica nanoparticles detached from the glassware walls by serial dilution and agitation through epitaxy. We compared the biological activity, electrical current and silicon microparticle content (by means of scanning electron microscopy/energy-dispersive X-ray spectroscopy) of high dilutions (HDs) of arsenic prepared in plastic and glass vials to investigate the role of silica in their biological effects in vitro. MATERIALS AND METHODS: Co-cultures of macrophages and yeast (Saccharomyces cerevisiae) were treated with different HDs of arsenic prepared in plastic and glass vials. Macrophage morphology, phagocytosis index, nitric oxide (NO), and cytokine production were evaluated. RESULTS: Measurable amounts of silicon microparticles were detected only in the HDs prepared in glass vials, but ultra-centrifugation eliminated them. Specific and non-specific results were observed. Non-specific pro-inflammatory effects were seen in all dilutions prepared in plastic vials, including elevation of pro-inflammatory cytokines, NO and macrophage phagocytic index. Only the 200th centesimal dilution of arsenic produced specific decrease in interleukin-6 production in macrophages, and it was independent of the vial type or the presence of microparticles of silica in the medicine samples. The nature of the vials had an impact on the electric flow in the respective fluids. CONCLUSION: The non-specific, pro-inflammatory effects might be attributed to organic residuals detached from the vials' plastic walls during manipulation. Instead, specific silica-independent effects of the homeopathic medicine can be attributed to the decrease of interleukin-6 after treatment with the 200th centesimal dilution of arsenic.

Homeopathic medicine Cantharis modulates uropathogenic E. coli (UPEC)-induced cystitis in susceptible mice.

OBJECTIVE: This is a random blinded placebo controlled murine experimental model to study the effects of Cantharis 6 CH, a homeopathic medicine, on E coli-induced cystitis. METHODS: 24 adult susceptible female BALB/c mice were inoculated with E coli - UPEC O4:K-:H5 by a transurethral catheter. Cantharis 6cH or vehicle (placebo) was offered to mice by free access into the drinking water (1:100), during 24 h after infection. Spleen, bladder and kidneys were processed for quantitative histopathology after immunohistochemistry, using anti-CD3, CD79, MIF, NK and VEGF antibodies; the cytokines present in the bladder washing fluid were measured using a LUMINEX-Magpix KIT. Mann-Whitney and Fisher exact test were used as statistical analysis. RESULTS: Cantharis 6 CH increased IL12p40, IFN-γ and decreased IL10 concentrations in the bladder fluid (p⩽0.05); in the bladder mucosa, it increased the ratio between B and T lymphocytes (31%) and between B lymphocytes and MIF+ macrophages (57%, p⩽0.05). In the pelvis, instead, it decreased the B/T cells ratio (41%, p⩽0.05) and increased the M1/M2 macrophage ratio (42%, p⩽0.05). No differences were seen in the kidney and spleen analysis. CONCLUSION: The inverted balance of inflammatory cells and cytokines in bladder and pelvis mucosa shows specific local immune modulation induced by Cantharis 6cH.

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)