232 resultados para In vivo assay


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Pós-graduação em Ciência Odontólogica - FOA

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Purpose: The antitumor activity of Kielmeyera coriacea (Clusiaceae), a medicinal plant used in the treatment of parasitic, as well as fungal and bacterial infections by the Brazilian Cerrado population, was investigated. Methods: A chloroform extract (CE) of K. coriacea was tested in the murine melanoma cell line (B16F10-Nex2) and a panel of human tumor cell lines. Tumor cell migration was determined by the wound-healing assay and the in vivo antitumor activity of CE was investigated in a melanoma cell metastatic model. 1H NMR and GC/MS were used to determine CE chemical composition. Results: We found that CE exhibited strong cytotoxic activity against murine melanoma cells and a panel of human tumor cell lines in vitro. CE also inhibited growth of B16F10- Nex2 cells at sub lethal concentrations, inducing cell cycle arrest at S phase, and inhibition of tumor cell migration. Most importantly, administration of CE significantly reduced the number of melanoma metastatic nodules in vivo. Chemical analysis of CE indicated the presence of the long chain fatty compounds, 1-eicosanol, 1-docosanol, and 2-nonadecanone as main constituents. Conclusion: These results indicate that K. coriacea is a promising medicinal plant in cancer therapy exhibiting antitumor activity both in vitro and in vivo against different tumor cell lines.

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Objective: To compare the efficacy of the mouthwashes 0.12% chlorhexidine, Listerine, and 0.5% and 2% Melaleuca Alternifolia oil against the salivary levels of Streptococcus mutans and total microorganisms. Methods: This study was double-blind controlled and paired clinical assay. Twenty-six volunteers aged 21 to 35 years old were enrolled. At baseline, 1 mL of unstimulated saliva was collected from each subject, 1 and 15 min after mouthrinsing with the following solutions: sterile distilled water, 0.12% chlorhexidine digluconate, Listerine (©Johnson & Johnson do Brasil), 0.5% and 2% concentrations of Melaleuca Alternifolia (Sigma-Aldrich). The volunteers used all the evaluated mouthrinses with a 15-day interval between the solutions. Immediately after rinsing, saliva was collected and serial dilutions were performed, followed by plating in blood agar culture medium for growth of total microorganisms and SB-20 (Sucrose-Bacitracin agar) for growth of S. mutans, and incubation at 37 °C for 48 h in microaerophilia. After incubation, the number of colonies was counted and expressed as colony forming units (UFC/mL). Results: Chlorhexidine showed antimicrobial action by reducing total microorganisms and S. mutans, while the action of 0.5% Melaleuca Alternifolia was similar to that of distilled water. Listerine and 2% Melaleuca Alternifolia oil reduced total microbial counts by 11% and 9% respectively, and S. mutans by 20% and 11%. Conclusion: A single rinse with 0.12% chlorhexidine is effective in reducing the levels of total microorganisms and S. mutans present in saliva. Under the same testing conditions, Listerine and 0.5% and 2% Melaleuca Alternifolia oil presented lower efficacy than chlorhexidine.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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The potential sequelae of intestinal infection with Yersinia enterocolitica include reactive arthritis, erythema nodosum, Reiter's syndrome and other autoimmune diseases. The role of the immune response in the pathogenesis of these diseases has not been fully defined, but autoimmune manifestations may be a consequence of the increase in autoantibodies as a result of polyclonal B-cell activation induced by Yersinia. We investigated the effects of Y enterocolitica 0:3 derivatives on B lymphocyte activation in vivo. Groups of five specific pathogen free (SPF) Swiss mice were inoculated with bacterial cell extract, Yersinia outermembrane proteins (Yops) or lipopolysaccharide (LPS) obtained from Y enterocolitica 0:3 and their immunoglobulin-secreting spleen cells were detected by isotype-specific protein A plaque assay. The presence of specific anti-Yersinia antibodies and autoantibodies was determined in mouse sera by ELISA. In all experiments a marked increase in the number of secretory cells of different isotypes was observed as early as the third day after inoculation. IgG and IgM anti-Yersinia antibodies were detected in the sera of all inoculated mice, and autoantibodies against myosin in the sera of those inoculated with bacterial cell extract. The sera from animals stimulated with LPS reacted with myelin, actin and laminin, while the sera from mice inoculated with Yops reacted with myelin, thyroglobulin and cardiolipin. These results suggest that SPF Swiss mice inoculated with any one of the Y enterocolitica derivatives tested exhibited polyclonal activation of B lymphocytes as a result of stimulation by various bacterial components and not only LPS stimulation.

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The treatment of schistosomiasis depends on a single drug: praziquantel (PZQ). However, this treatment presents limitations such as low and/or erratic bioavailability that can contribute to cases of tolerance. Improvements to the available drug are urgently needed and studies with a controlled system of drug release, like liposomes, have been gaining prominence. The present study evaluated the activity and synergy between liposomal-praziquantel (lip.PZQ) and hyperbaric oxygen therapy (HBO). Mice received doses of 60 or 100mg/kg PZQ or lip.PZQ, 50 days post-infection, and after the treatment, were exposed to HBO (3 atmosphere absolute - ATA) for 1h. The viability of adult worms and oviposition were analyzed, by necropsy and Kato-Katz examination performed after 15 days of treatment. A concentration of 100mg/kg of lip.PZQ+HBO was more effective (48.0% reduction of worms, 83.3% reduction of eggs/gram of feces) and 100% of the mice had altered of oograms (indicating interruption of oviposition) compared to other treatments and to the Control group (infected and untreated). It is known that PZQ requires participation of the host immune system to complete its antischistosomal activity and that HBO is able to stimulate the immune system. The drug became more available in the body when incorporated into liposomes and, used with HBO, the HBO worked as an adjuvant. This explains the decreases of oviposition and worms recovered form hepatic portal system.

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It is know that endotoxin and various matrix metalloproteinases (MMPs) are involved in the development of periapical lesions. The purpose of this study was to evaluate and correlate the presence of endotoxins and MMP- 3, MMP-8 and MMP-9 in root canals of teeth with necrotic pulp and periapical lesion before, during and after the biomechanical preparation (PBM) using a combination of different irrigations solutions and intracanal dressing. Thirty-three single-root teeth with a diagnosis of pulp necrosis and periapical lesion radiographically visible were selected. Immediately after the coronal opening was collected the first sample from the root canal content. Then, all canals were prepared (cervical and middle thirds) by oscillatory instruments (EndoEze) and irrigated by 2.5% NaOCl. After, a manual preparation was made for the apical third and the teeth were divided into three groups according to the irrigation protocol: G1) 2.5% NaOCl (4 manual files); G2) 2.5% NaOCl (2 manual files) + [Ca (OH)2 0.14%] (2 manual files) and G3) 2.5% NaOCl (2 manual files) + polymyxin B (2 manual files). After the PBM, the second sample was collected; then the third collect was performed after using EDTA final flush. The fourth sample was collected 14 days after placing the dressing [2% chlorhexidine gel + Ca(OH)2]. Quantification of endotoxins was performed by a kinetic chromogenic lysate from amoebocytes of Limulus (LAL) and quantification of MMPs by ELISA assay. The results were analyzed statistically by Kruskal-Wallis and Dunn's test (5%) and ordinal Spearman correlation. Presence of endotoxin was observed in 100% of cases and G3 showed the greatest reduction of endotoxins from the 1st to the 2nd samples (97%), being statistically similar to G2 (84.2%) and different from G1 (49.4%) (p<0.05). The intracanal dressing promoted a significant reduction of endotoxin, no difference among the groups. For...

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Background The recent emergence of extensively multidrug-resistant Mycobacterium tuberculosis strains has further complicated the control of tuberculosis. There is an urgent need for the development of new molecular candidates antitubercular drugs. Medicinal plants have been an excellent source of leads for the development of drugs. The aim of this study was to evaluate the in vitro activity of 28 alcoholic extracts and essential oils of native and exotic Brazilian plants against Mycobacterium tuberculosis and to further study these extracts through chemical fractionation, the isolation of their constituents, and an evaluation of the in vivo acute toxicity of the active extracts. To the best of our knowledge this is the first chemical characterization, antituberculosis activity and acute toxicity evaluation of Annona sylvatica. Methods The anti-mycobacterial activity of these extracts and their constituent compounds was evaluated using the resazurin reduction microtiter assay (REMA). To investigate the acute toxicity of these extracts in vivo, female Swiss mice were treated with the extracts at doses of 500, 1000 and 2000 mg · kg-1 of body weight. The extracts were characterized by LC-MS, and the constituents were isolated and identified by chromatographic analysis of spectroscopic data. Results Of the 28 extracts, the methanol extract obtained from the leaves of Annona sylvatica showed anti-mycobacterial activity with an minimal inhibitory concentration (MIC) of 184.33 μg/mL, and the ethyl acetate fraction (EAF) resulting from liquid-liquid partitioning of the A. sylvatica extract showed an MIC of 115.2 μg/mL. The characterization of this extract by LC-MS identified flavonoids and acetogenins as its main constituents. The phytochemical study of the A. sylvatica EAF resulted in the isolation of quercetin, luteolin, and almunequin. Conclusions Among the compounds isolated from the EAF, luteolin and almunequin were the most promising, with MICs of 236.8 μg/mL (827.28 μM) and 209.9 μg/mL (328.48 μM), respectively. The acute administration of the EAF fraction in doses of 500, 1000, and 2000 mg · kg-1 of body weight did not cause signs of toxicity in the treated animals.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)