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The present study analyzed the immune modulation mechanisms of thymulin 5CH in a granuloma experimental model. Male adult Balb/c mice were inoculated with BCG into the footpad to induce granuloma, which was quantitatively evaluated. The phenotypic characterization of phagocyte, T- and B-lymphocyte populations in the peritoneum, and local lymph node was done by flow cytometry. During all experimental periods, thymulin 5CH and vehicle (control) were given ad libitum to mice, diluted into the drinking water (1.6 × 10(-17) M). After 7 days from inoculation, thymulin-treated mice presented reduction in the number of epithelioid cytokeratine-positive cells (P = 0.0001) in the lesion, in relation to young phagocytes. After 21 days, the differentiation of B1 peritoneal stem cells into phagocytes reached the peak, being higher in thymulin-treated mice (P = 0.0001). Simultaneously, the score of infected phagocytes in the lesion decreased (P = 0.001), and the number of B1-derived phagocytes, CD4+ and CD8+ T lymphocytes in the local lymph node increased in relation to control (P = 0.0001). No difference was seen on the CD25+ Treg cells. The results show that thymulin 5CH treatment is able to improve the granuloma inflammatory process and the infection remission, by modulating local and systemic phagocyte differentiation.

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As little information about the effect of ultra high dilutions of glucocorticoid in reproduction is available in the literature, pregnant female Wistar rats (N = 12) were blindly subcutaneously treated during all gestational and lactation period with: dexamethasone 4 mg/kg diluted into dexamethasone 15 cH (mixed); or dexamethasone 4 mg/kg diluted in water; or dexamethasone 15 cH, or vehicle. Parental generation had body weight, food and water consumption monitored. The F1 generation was monitored regarding to newborn development. No birth occurred in both groups treated with dexamethasone 4 mg/kg. After 60 days from birth, 12 male F1 rats were randomly selected from each remaining group and inoculated subcutaneously with 1% carrageenan into the footpad, for evaluation of inflammatory performance. Edema and histopathology of the footpad were evaluated, using specific staining methods, immunohistochemistry and digital histomorphometry. Mothers treated with mixed dexamethasone presented reduced water consumption. F1 rats born to dexamethasone 15 cH treated females presented significant increase in mast cell degranulation, decrease in monocyte percentage, increase in CD18+ PMN cells, and early expression of ED2 protein, in relation to control. The results show that the exposure of parental generation to highly diluted dexamethasone interferes in inflammation modulation in the F1 generation.

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Modulation of immune response due to thymulin 5cH has been previously observed. The aim of the present study is to evaluate the development of broiler chickens treated with thymulin 5cH by conventional zoo technical indices, phytohemaglutinin induced inflammation test and histomorphometric analysis of lymphoid organs (thymus, Fabricius bursa and spleen). Animals were divided in two groups: (a) test: birds with free access to thymulin 5cH diluted into the drink water and (b) control: birds with free access to water only, from the 1st to the 42nd day of life. All experimental procedures were done in blind. The results show that thymulin 5cH treated group had increased productivity index compared to control (391.45 versus 261.93) associated with higher viability in the 7th week (p = 0.013), and a possible shunt to B lymphocyte activity. The data suggest that thymulin 5cH could be a viable method to improve productivity in poultry production due to its immune modulation properties.

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Em estudos prévios, foi verificado que a timulina (um hormônio tímico), quando preparada em potências homeopáticas 5cH, tem a propriedade de melhorar a performance produtiva de frangos infectados com reovirus, bem como modular o desenvolvimento do tumor de Ehrlich e de lesões granulomatosas em camundongos por mecanismos imunomediados. O objetivo deste trabalho foi estudar os mecanismos imunonodulatórios da timulina 5cH em um modelo de granuloma experimental, pela injeção subcutânea de BCG em camundongos, focando o envolvimento das células B1 e do zinco no processo. Três grupos de camundongos Balb/c machos (grupo A tratado com timulina 5cH, grupo B tratado com timulina 5cH incubada com Chelex ® - um quelante de zinco ? e grupo C, controle, tratado com veículo) foram inoculados com BCG no coxim plantar esquerdo e o granuloma subcutâneo e baço foram colhidos para análise histomorfométrica, após 7, 14 e 21 dias. Colorações de Ziehl-Neelsen, hematoxilina eosina e azul da Prússia foram também utilizadas. A citometria de fluxo também foi usada nos mesmos tempos para caracterizar e quantificar células peritoneais. Células positivas para CD11b (fagócitos ativados, células B1), CD 19 (B1 e B2), CD23 (negativas em B1, positivas em B2) e CD5 (B1a) foram analisadas em um aparelho FACS Calibur (BD). A análise estatística foi Kruskal-Wallis/Dunn para avaliações não paramétricas e ANOVA/Tuckey-Krammer para avaliações paramétricas. O método X2 foi usado para avaliar a proporção entre células na citometria. Valores de p?0.05 foram considerados estatisticamente significantes. Camundongos tratados com timulina 5cH apresentaram alta atividade de macrófagos e aumento na área folicular esplênica após 7 dias da inoculação do BCG. O aumento do tamanho macroscópico da lesão e a redução da infecção local foram vistos após 21 dias. Nesse período, a citometria demonstrou aumento na proporção de fagócitos derivados de células B1 peritoneais em camundongos tratados com timulina 5cH, independentemente da incubação ou não com Chelex®, a qual bloqueou apenas os efeitos da timulina sobre células B2 no peritôneo e reduziu significativamente os níveis de manganês na preparação medicamentosa. Conclui-se que a timulina 5cH modula o granuloma induzido por BCG através de mais de um mecanismo, especialmente da diferenciação de células B1 peritoneais em fagócitos.(AU)

In previous studies, we found that thymulin (a thymic hormone), when prepared in homeopathic 5cH potency, had the property to improve the productive performance of broiler chickens infected with reovirus, as well as modulate the development of Ehrlich tumor and granuloma inflammatory lesions in mice by immune-mediated mechanisms. The aim of the present work was to study the immunomodulatory mechanisms of thymulin 5cH in a granuloma experimental model, by subcutaneous inoculation of BCG in mice, focusing the B-1 cells and zinc involvement in this process. Three groups of male Balb/c SPF mice (group A treated with thymulin 5cH, group B treated with thymulin 5cH incubated in Chelex ® - a zinc chelant - and group C, control, treated with vehicle) were inoculated with BCG in the left footpad and subcutaneous granuloma and spleen were harvested for histomorphometry analysis, after 7, 14 and 21 days. Ziehl-Neelsen, HE and Prussia Blue staining methods were used. Flow cytometry was also used in the same times to characterize and quantify peritoneal cells. Positive cells for CD11b (activated phagocytes, B-1 cells), CD19 (B-1 and B-2 cells), CD23 (negative B-1 cells, positive B2 cells) and CD5 (B-1a cells) were analyzed in a FACS Calibur (BD) device. Statistical analysis was performed using Kruskal - Wallis / Dunn for nonparametric evaluations and ANOVA / Tuckey-Krammer for the parametric ones. The X² method was used to evaluate the cell count in flow cytometry. P values ? 0.05 were considered statistically significant. Mice treated with thymulin 5cH presented higher macrophage activity and increase in the follicular area were seen in spleen after 7 days. Increase in gross lesion diameter and decrease in local BCG infection were seen after 21 days. At this time, the flow cytometry demonstrated the increase in peritoneal phagocytes derived from B-1 cells in thymulin 5cH treated mice, independently of Chelex ® incubation. The incubation of thymulin 5cH with Chelex ® blocked its effects only upon the number of B2 cells in the peritoneum and reduces Mn levels in the medicine solution. We conclude that thymulin 5CH modulates the BCG-induced granuloma through more than one mechanism, especially by peritoneal B1 cell differentiation into phagocytes.(AU)

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The effects of Arnica montana 6cH on the individual modulation of acute inflammation kinetics in rats were evaluated. Adult male Wistar rats were inoculated with 1% carrageenan into the footpad and treated with Arnica montana 6cH, dexamethasone (4.0 mg/kg; positive control) or 5% hydroalcoholic solution (negative control), per os, each 15 minutes, between 30 and 180 minutes after the irritant inoculation. Histopathological and immunohistochemistry procedures were done in order to get a panel of inflammatory positive cells for CD3 (T lymphocytes), CD45RA (B lymphocytes), CD18 (beta 2 integrin), CD163 (ED2 protein), CD54 (ICAM-1), and MAC 387 (monocytes and macrophages). The statistical treatment of data included a posteriori classification of animals from each group (N = 20) in two subgroups presenting spontaneous precocious or late oedema. Animals that presented precocious oedema were less responsible to Arnica montana 6cH in relation to hemodynamic changes. Instead, rats that exhibited late oedema presented less intense oedema (P = .01), lower percentage of mast cell degranulation (P = .0001), and increase in lymphatic vessels diameter (P = .05). The data suggest an individually qualitative adjustment of inflammatory vascular events by Arnica montana 6cH.