Effect of a submaxillary gland extract on Ehrlich tumor growth in mice

Ablation of host submaxillary glands modifies Ehrlich tumor growth and tumor-infiltrating leukocytes, possibly by modifications in the serum level of growth factors produced by this gland. To extend this research, 7-month-old male EPM-1 mice (N = 30) were divided into two groups: 1) inoculated with tumor cells previously incubated with submaxillary salivary gland extract (SGE) in PBS for 30 min at 37%; 2) inoculated with tumor cells previously incubated with PBS, under the same conditions. Animals were inoculated into the footpad with 40 µl of a suspension containing 4.5 x 107 tumor cells/ml, and footpad thickness was measured daily for 10 days. Sections and smears of tumor cells were prepared from the tumor mass to determine mitosis frequency, percent of tumor cells immunopositive to nerve (NGF) and epidermal (EGF) growth factors and percent of tumor-infiltrating leukocytes. The incubation of tumor cells with SGE produced a tumor reduction of about 30% in size (P<0.01). This effect was not related to loss of cell viability during incubation, but a 33% increase (P<0.05) in the percentage of dead or dying tumor cells and a 15% increase in the percent of NGF/EGF-positive tumor cells (P<0.01) were observed in vivo at the end of experiment. Tumor-infiltrating lymphocytes and mitosis frequency did not differ between groups. These data suggest a direct effect of factors present in SGE on tumor cells, which induce degeneration of tumor cells.Universidade PaulistaUniversidade Federal de São Paulo (UNIFESP)UNIFESPSciEL

Inflammatory Process Modulation by Homeopathic Arnica montana 6CH: The Role of Individual Variation

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

Embriotoxic effects of Crotalus durissus terrificus snake venom

A atividade neurotóxica, miotóxica e coagulante do veneno das serpentes Crotalus durissus terrificus (VCdt) são responsáveis pelas altas taxas de mortalidade observada em acidentes envolvendo estas serpentes. Estes acidentes, quando ocorrem na gravidez, podem levar ao aborto devido à interferência com a homeostasia materna e/ou à embrioletalidade, por efeito direto do veneno. Materiais e Métodos - Este trabalho estudou os efeitos tóxicos do ( VCdt), administrado nas doses de 75 mg/kg or 200 mg/kg por via subcutânea em camundongas no terceiro dia da sua gestação (período de pré-implantação). O grupo controle foi tratado da mesma forma que os experimentais, porém com solução salina. No último dia da gestação as fêmeas foram submetidos a eutanásia e observadas as possíveis malformações ósseas e viscerais de sua prole. Resultados e Conclusão - Os resultados mostraram que a administração da menor dose do VCdt não causou alterações significantes no desenvolvimento ósseo e visceral dos animais. No entanto, quando expostos a maior dose este promoveu aumento significante das anomalias e malformações, sugerindo que o envenenamento com esta dose no período inicial da gestação altera o desenvolvimento normal da prole de camundongosIntroduction - The neurotoxic, myotoxic and coagulant activities of Crotalus durissus terrificus snake venom (VCdt) are responsible for the mortality rates observed in accidents involving the rattlesnake. Accidents during women pregnancy are a challenge, since animal venoms could led to pregnancy interruption as a consequence of maternal homeostasis disorder and/or a direct embryotoxic effect of the venom. Materials and Methods - In order to evaluate the possible embryotoxic effects of VCdt, doses of 75 mg/kg or 200 mg/kg of the venom were administered by subcutaneous route at day 3 of mice pregnancy (preimplantation period). The control group received saline in the same volume and during the same period as their respective experimental groups. The animals were submitted to euthanasia at term. Results and Conclusion - The treatment of the females during the preimplantation period did not cause significant changes in fetuses development, but the higher dose of the venom increased the number of anomalies or malformations of the fetuses. These results suggest that the VCdt in the higher dose (200 mg/kg) altered the normal development of the concept after implantatio

Highly Diluted Glyphosate Mitigates Its Effects on Artemia salina: Physicochemical Implications.

Glyphosate is an herbicide widely used in agriculture but can present chronic toxicity in low concentrations. Artemia salina is a common bio-indicator of ecotoxicity; it was used herein as a model to evaluate the effect of highly diluted-succussed glyphosate (potentized glyphosate) in glyphosate-based herbicide (GBH) exposed living systems. Artemia salina cysts were kept in artificial seawater with 0.02% glyphosate (corresponding to 10% lethal concentration or LC10) under constant oxygenation, luminosity, and controlled temperature, to promote hatching in 48 h. Cysts were treated with 1% (v/v) potentized glyphosate in different dilution levels (Gly 6 cH, 30 cH, 200 cH) prepared the day before according to homeopathic techniques, using GBH from the same batch. Controls were unchallenged cysts, and cysts treated with succussed water or potentized vehicle. After 48 h, the number of born nauplii per 100 µL, nauplii vitality, and morphology were evaluated. The remaining seawater was used for physicochemical analyses using solvatochromic dyes. In a second set of experiments, Gly 6 cH treated cysts were observed under different degrees of salinity (50 to 100% seawater) and GBH concentrations (zero to LC 50); hatching and nauplii activity were recorded and analyzed using the ImageJ 1.52, plug-in Trackmate. The treatments were performed blind, and the codes were revealed after statistical analysis. Gly 6 cH increased nauplii vitality (p = 0.01) and improved the healthy/defective nauplii ratio (p = 0.005) but delayed hatching (p = 0.02). Overall, these results suggest Gly 6cH treatment promotes the emergence of the more GBH-resistant phenotype in the nauplii population. Also, Gly 6cH delays hatching, another useful survival mechanism in the presence of stress. Hatching arrest was most marked in 80% seawater when exposed to glyphosate at LC10. Water samples treated with Gly 6 cH showed specific interactions with solvatochromic dyes, mainly Coumarin 7, such that it appears to be a potential physicochemical marker for Gly 6 cH. In short, Gly 6 cH treatment appears to protect the Artemia salina population exposed to GBH at low concentrations

Comfrey (Symphytum Officinale. l.) and Experimental Hepatic Carcinogenesis: A Short-term Carcinogenesis Model Study

Comfrey or Symphytum officinale (L.) (Boraginaceae) is a very popular plant used for therapeutic purposes. Since the 1980s, its effects have been studied in long-term carcinogenesis studies, in which Comfrey extract is administered at high doses during several months and the neoplastic hepatic lesions are evaluated. However, the literature on this topic is very poor considering the studies performed under short-term carcinogenesis protocols, such as the ‘resistant hepatocyte model’ (RHM). In these studies, it is possible to observe easily the phenomena related to the early phases of tumor development, since pre-neoplastic lesions (PNLs) rise in about 1–2 months of chemical induction. Herein, the effects of chronic oral treatment of rats with 10% Comfrey ethanolic extract were evaluated in a RHM. Wistar rats were sequentially treated with N-nitrosodiethylamine (ip) and 2-acetilaminofluorene (po), and submitted to hepatectomy to induce carcinogenesis promotion. Macroscopic/microscopic quantitative analysis of PNL was performed. Non-parametric statistical tests (Mann–Whitney and χ2) were used, and the level of significance was set at P ≤ 0.05. Comfrey treatment reduced the number of pre-neoplastic macroscopic lesions up to 1 mm (P ≤ 0.05), the percentage of oval cells (P = 0.0001) and mitotic figures (P = 0.007), as well as the number of Proliferating Cell Nuclear Antigen (PCNA) positive cells (P = 0.0001) and acidophilic pre-neoplastic nodules (P = 0.05). On the other hand, the percentage of cells presenting megalocytosis (P = 0.0001) and vacuolar degeneration (P = 0.0001) was increased. Scores of fibrosis, glycogen stores and the number of nucleolus organizing regions were not altered. The study indicated that oral treatment of rats with 10% Comfrey alcoholic extract reduced cell proliferation in this model

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

ABSTRACT 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

Advance Access Publication

Comfrey or Symphytum officinale (L.) (Boraginaceae) is a very popular plant used for therapeutic purposes. Since the 1980s, its effects have been studied in long-term carcinogenesis studies, in which Comfrey extract is administered at high doses during several months and the neoplastic hepatic lesions are evaluated. However, the literature on this topic is very poor considering the studies performed under short-term carcinogenesis protocols, such as the &apos;resistant hepatocyte model&apos; (RHM). In these studies, it is possible to observe easily the phenomena related to the early phases of tumor development, since pre-neoplastic lesions (PNLs) rise in about 1-2 months of chemical induction. Herein, the effects of chronic oral treatment of rats with 10% Comfrey ethanolic extract were evaluated in a RHM. Wistar rats were sequentially treated with N-nitrosodiethylamine (ip) and 2-acetilaminofluorene (po), and submitted to hepatectomy to induce carcinogenesis promotion. Macroscopic/microscopic quantitative analysis of PNL was performed. Non-parametric statistical tests (Mann-Whitney and 2 ) were used, and the level of significance was set at P 0.05. Comfrey treatment reduced the number of pre-neoplastic macroscopic lesions up to 1 mm (P 0.05), the percentage of oval cells (P = 0.0001) and mitotic figures (P = 0.007), as well as the number of Proliferating Cell Nuclear Antigen (PCNA) positive cells (P = 0.0001) and acidophilic pre-neoplastic nodules (P = 0.05). On the other hand, the percentage of cells presenting megalocytosis (P = 0.0001) and vacuolar degeneration (P = 0.0001) was increased. Scores of fibrosis, glycogen stores and the number of nucleolus organizing regions were not altered. The study indicated that oral treatment of rats with 10% Comfrey alcoholic extract reduced cell proliferation in this model

Modulation of inflammation response to murine cutaneous Leishmaniasis by homeopathic medicines: Antimonium crudum 30cH

Background: Leishmaniasis is a zoonotic disease caused by protozoan parasites of the mononuclear phagocytic system. the modulation activity of these cells can interfere in the host/parasite relationship and influences the prognosis.Methods: We evaluated the effects of the homeopathic preparation Antimonium crudum 30cH on experimental infection induced by Leishmania (L.) amazonensis. Male Balb/c mice were inoculated with 2 x 10(6) Leishmania (L.) amazonensis promastigotes into the footpad and, after 48 h (acute phase) or 60 days (chronic phase), cell population of lymphocytes and phagocytes present in the peritoneal washing fluid and spleen were analyzed by flow cytometry and histopathology, with histometry of the subcutaneous primary lesion, local lymph node and spleen. Immunohistochemistry was performed to quantify CD3 (T lymphocyte), CD45RA (B lymphocyte) and CD11b (phagocytes) positive cells.Results: in treated mice, during the acute phase, there was significant increase of the macroscopic lesion, associated to inflammatory edema, as well increase in the number of free amastigotes and B lymphocytes inside the lesion. Increase of B lymphocytes (predominantly B-2 cells) was also seen in the local lymph node, spleen and peritoneum. in the chronic phase, the inflammatory process in the infection focus was reduced, with reduced phagocyte migration and peritoneal increase of B-1a cells (precursors of B-2 immunoglobulin producers cells) and T CD8+ cells.Conclusion: the treatment of mice with Antimonium crudum 30cH induced a predominantly B cell pattern of immune response in Leishmania (L.) amazonensis experimental infection, alongside the increase of free amastigote forms number in the infection site. the clinical significance of this study is discussed, further studies are suggested.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)UNIPCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Univ Paulista, Res Ctr, Grad Program Environm & Expt Pathol, BR-04026002 São Paulo, BrazilUniv Santo Amara, Lab Vet Pathol, São Paulo, BrazilUniversidade Federal de São Paulo, Immunol Lab, São Paulo, BrazilUniv São Paulo, Vet & Zootechny Fac, BR-05508 São Paulo, BrazilUniv São Paulo, Fac Med, São Paulo, BrazilUniversidade Federal de São Paulo, Immunol Lab, São Paulo, BrazilFAPESP: 2010/50842-0Web of Scienc

Efeito da Avena sativa CH6 no metabolismo do sêmen diluído de suínos

Este trabalho avaliou a utilização da Avena sativa no sêmen diluído de reprodutores suínos com o objetivo de melhorar a atividade metabólica das células espermáticas. O experimento foi efetuado em uma granja comercial de suínos com 200 fêmeas. Foram formados dois grupos, tratado e controle, com sesenta fêmeas reprodutoras em cada um. O sêmen dos reprodutores machos foi originário de dois animais sexualmente maduros e foi avaliada a motilidade, concentração e vigor espermático em zero, 24, 48, 72 e 96 horas pós colheita. No grupo tratado foi adicionado Avena sativa CH6 (dois glóbulos) por litro de água destilada adicionados ao diluente. Para o grupo controle foi adicionado somente diluente. Foi detectada diferença estatística nas 48 e 96 horas pós colheita do sêmen (p<0.05) com maior motilidade espermática no grupo tratado. Em relação ao vigor espermático foi detectada diferença estatística nas 48 e 72 horas pós colheita do sêmen (p<0.05) com maior vigor no grupo tratado. O número de retornos ao cio maior no grupo controle (20.00%) em comparação ao tratado (16,66%) sem diferença estatisticamente significativa. No grupo controle nasceram 464 leitões e no tratado, 516. A inclusão da Avena sativa no sêmen diluído contribuiu para melhorar a atividade metabólica dos espermatozóides com mairo motilidade e vigor espermáticoThe use of Avena sativa prepared homeopathically on diluted semen of boars was studied in order to verify a putative improvement in the sperm cells metabolic activity. The experiment was developed in a commercial swine herd composed by 200 sows. Two groups were divided, named treatd and control, with sixty sows in each. Semen was collected from two boars and motility, concentration and sperm vigor were evaluated at zero, 24, 48, 72 and 96 hours after collection. For treated group it was added Avena sativa CH6 (two globules) per liter of distilled water added to dilluent. For control group it was added only dilluent. Treated group showed statistical significant difference in sperm motility at 48 and 96 hours after semen collection (P<0.05). In relation to spem vigor, statistical difference was seen after 48 and 72 hours (p<0.05) for the treated group. The number of return to estrus was higher at the control group (20.00%) compared to treated group (16.66%) without statistical significance. A total of 464 piglets were born in control group and 516 were born in treated group. The inclusion of globules of Avena sativa 6CH on diluted semen contributed to improve sperm metabolic activity with higher motility and sperm cells vigo