Myotoxic phospholipases A2 isolated from Bothrops brazili snake venom and synthetic peptides derived from their C-terminal region: Cytotoxic effect on microorganism and tumor cells

This paper reports the purification and biochemical/pharmacological characterization of two myotoxic phospholipases A2 (PLA2s) from Bothrops brazili venom, a native snake from Brazil. Both myotoxins (MTX-I and II) were purified by a single chromatographic step on a CM-Sepharose ion-exchange column up to a high purity level, showing Mr ∼ 14,000 for the monomer and 28,000 Da for the dimer. The N-terminal and internal peptide amino acid sequences showed similarity with other myotoxic PLA2s from snake venoms, MTX-I belonging to Asp49 PLA2 class, enzymatically active, and MTX-II to Lys49 PLA2s, catalytically inactive. Treatment of MTX-I with BPB and EDTA reduced drastically its PLA2 and anticoagulant activities, corroborating the importance of residue His48 and Ca2+ ions for the enzymatic catalysis. Both PLA2s induced myotoxic activity and dose-time dependent edema similar to other isolated snake venom toxins from Bothrops and Crotalus genus. The results also demonstrated that MTXs and cationic synthetic peptides derived from their 115-129 C-terminal region displayed cytotoxic activity on human T-cell leukemia (JURKAT) lines and microbicidal effects against Escherichia coli, Candida albicans and Leishmania sp. Thus, these PLA2 proteins and C-terminal synthetic peptides present multifunctional properties that might be of interest in the development of therapeutic strategies against parasites, bacteria and cancer. © 2008 Elsevier Inc. All rights reserved.

Comparison of exfoliative pap stain and AgNOR counts of the tongue in smokers and nonsmokers

Objective To compare exfoliative cytology from the oral mucosa of smokers and nonsmokers, with emphasis on proliferative activity. Methods Exfoliative cytology specimens were obtained from clinical normal mucosa from the lateral border of the tongue in 30 nonsmokers and 30 smokers ranging in age from 40 to 70 years of age, who were seen at the Heart Institute's Patient Center and the Smoking Cessation Program of the University Hospital, University of São Paulo Medical School (InCor-HCFMUSP). The cytologic specimens were evaluated by Papanicolaou staining and AgNOR quantification in order to evaluate the presence of cytological alterations suggestive of inflammation, dysplasia, keratinization, and proliferative activity of epithelial cells. Results Only Papanicolaou Class I and Class II smears were observed. Inflammatory alterations were found in 90% of smokers and in 87% of nonsmokers. The number of AgNORs/nucleus differed significantly between smokers and nonsmokers (3.372 ± 0.375 versus 2.732 ± 0.236). Conclusions Within the limitations of this research, the results indicate higher proliferative activity in smoking patients compared to nonsmoking patients, even in the absence of clinical lesions. © The Author(s) 2008.

Atividade antioxidante e antimicrobiana do alecrim (Rosmarinus officinalis L.) em filés de tilápia (Oreochromis ssp) salgados secos durante o armazenamento congelado

Rosemary (Rosmarinus officinalis L.) is a spice from Lamiaceae family known since ancient times because its medicinal effects and, currently, several studies have pointed its antioxidant and antimicrobian effects. Lipid oxidation is a problem in food production because proceed the lost of organoleptical and nutritional qualities so required in Market. Fish salting is an ancient conservation method that expect reduce water activity and, consequently, microorganism growth in food, except halophillic bacteria. In the meantime, the inconvenient of this procedure is that the salt accelerates tissue's lipid oxidation. The aim of this work was evaluate the antioxidative and antimicrobian effects by treatment and pre treatment with rosemary (Rosmarinus officinalis L.) aqueous extract in dry salted tilapia fillets, storaged in freezing temperatures. To follow the oxidative, dry salted tilapia fillets were treated or pre treated with rosemary natural extract and storage at -18°C for 240 days. Analisys of 2-thiobarbituric acid reactive substances (TBARS), soluble nitrogen in trichloroacetic acid (TCA), water activity and microbiology were done. The pre treatment (3.39±0,53) and the treatment with rosemary (3.31±0.79) had oxidative index twice lower than the control treatment (6.14±1.21) in the last time of the research. The microbiological rosemary analisys showed count levels of resistant microorganisms to salt (2.0×103CFU/g of sample), whom causes the initial fillets contamination. The microbiological counts remained invariable in all groups during storage periods.

Influence of preventive dental treatment on mutans streptococci counts in patients undergoing head and neck radiotherapy

The aim of this study was to evaluate the influence of chlorhexidine gluconate, sodium fluoride and sodium iodine on mutans streptococci counts in saliva of irradiated patients. Forty-five patients were separated into three experimental groups and received chlorhexidine (0.12%), sodium fluoride (0.5%) or sodium iodine (2%), which were used daily during radiotherapy and for 6 months after the conclusion of the treatment. In addition, a fourth group, composed by 15 additional oncologic patients, who did not receive the mouthwash or initial dental treatment, constituted the control group. Clinical evaluations were performed in the first visit to dental clinic, after initial dental treatment, immediately before radiotherapy, after radiotherapy and 30, 60, 90 days and 6 months after the conclusion of radiotherapy. After clinical examinations, samples of saliva were inoculated on SB20 selective agar and incubated under anaerobiosis, at 37oC for 48 h. Total mutans streptococci counts were also evaluated by using real-time PCR, through TaqMan system, with specific primers and probes for S. mutans and S. sobrinus. All preventive protocols were able to reduce significantly mutans streptococci counts, but chlorhexidine gluconate was the most effective, and induced a significant amelioration of radiotherapy side effects, such as mucositis and candidosis. These results highlights the importance of the initial dental treatment for patients who will be subjected to radiotherapy for head and neck cancer treatment.

Evaluation of alternative methods for the disinfection of toothbrushes

The aim of this study was to evaluate alternative methods for the disinfection of toothbrushes considering that most of the previously proposed methods are expensive and cannot be easily implemented. Two-hundred toothbrushes with standardized dimensions and bristles were included in the study. The toothbrushes were divided into 20 experimental groups (n=10), according to microorganism considered and chemical agent used. The toothbrushes were contaminated in vitro by standardized suspensions of Streptococcus mutans, Streptococcus pyogenes, Staphylococcus aureus or Candida albicans. The following disinfectants were tested: 0.12% chlorhexidine digluconate, 50% white vinegar, a triclosan-containing dentifrice solution, and a perborate-based tablet solution. The disinfection method was immersion in the disinfectant for 10min. After the disinfection procedure, the number of remaining microbial cells was evaluated. The values of cfu/toothbrush of each group of microorganism after disinfection were compared by Kruskal-Wallis ANOVA and Dunn's test for multiple comparisons (5%). The chlorhexidine digluconate solution was the most effective disinfectant. The triclosan-based dentifrice solution promoted a significant reduction of all microorganisms' counts in relation to the control group. As to the disinfection with 50% vinegar, a significant reduction was observed for all the microorganisms, except for C. albicans. The sodium perborate solution was the less effective against the tested microorganisms. Solutions based on triclosan-containing dentifrice may be considered effective, nontoxic, cost-effective, and an easily applicable alternative for the disinfection of toothbrushes. The vinegar solution reduced the presence of S. aureus, S. mutans and S. pyogenes on toothbrushes.

Prevalência e sensibilidade aos antimicrobianos de microrganismos potencialmente superinfectantes na cavidade bucal de pacientes com espondilite anquilosante em uso de terapia anti TNF

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Salmonella spp. e Clostridium perfringens em farinhas de origem animal utilizadas na fabricação de rações e avaliação de aditivo na inibição de patógeno

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Presença de microrganismos potencialmente superinfactantes na cavidade bucal de pacientes com anemia falciforme em uso de hidroxiuréia

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação de substâncias desinfetantes utilizadas na água de tanques de pré-resfriamento de abatedouro avícola

Pós-graduação em Medicina Veterinária - FCAV

Efetividade de duas soluções desinfetantes e da irradiação por micro-ondas na desinfecção de próteses totais contaminadas com staphylococcus aureus resistente à meticilina (MRSA)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efetividade microbiológica da desinfecção por microondas e seus efeitos sobre a dureza de materiais reembasadores macios

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Recommendations for monitoring avian populations with point counts: a case study in southeastern Brazil

No hemisfério norte, o censo de aves é fundamental para gerar informações que auxiliam na compreensão de tendências populacionais. Tais censos, devido à marcada sazonalidade deste hemisfério, são realizados durante dois momentos distintos: na estação reprodutiva (aves residentes) e no inverno (quando as aves migratórias deixam determinadas regiões). Na região neotropical, porém, dependendo da localidade, as aves podem se reproduzir durante qualquer ou vários períodos do ano; podem ou não migrar, e aquelas que o fazem podem apresentar um padrão assincrônico. Em contraste com o hemisfério norte, tendências populacionais são desconhecidas, bem como o impacto das taxas rápidas de urbanização e desmatamento, que também são pouco monitoradas. Para melhor entender padrões temporais de riqueza e abundância de aves, e avaliar como um censo similar pode ser implementado na América tropical, foram utilizados pontos de escuta ao longo de 12 meses em uma localidade no Estado de São Paulo, sudeste do Brasil. Os censos ocorreram duas vezes por dia (manhãs/tardes) em uma floresta semidecidual ao longo de transecções com 10 pontos (20 pontos por dia) distantes 200 m entre si e com raio de detecção limitado em 100 m. Ambas as riquezas e abundâncias de aves foram maiores durante as manhãs, mas as curvas de acumulação sugerem que os censos vespertinos com maior esforço amostral podem fornecer resultados similares aos censos matutinos. Riqueza e abundância das aves não variam de acordo com estações (i.e., sem padrão aparente entre reprodução e migração), enquanto espécies exclusivas foram encontradas todos os meses e relativamente poucas espécies (20%) foram registradas em todos os meses do ano. Durante este ano, 84% de todas as aves florestais da área estudada foram registradas. Sugerimos que a metodologia de pontos de escuta pode ser utilizada à semelhança dos censos do hemisfério norte. Recomendamos ainda que o esforço amostral em transecções deva incluir ao menos 20 pontos, e que o início da contagem das aves deva ser sazonal, utilizando o período de migração das espécies austrais (e os seis meses seguintes) para coordenar pontos de escuta. Por último, sugerimos que os censos no Brasil e até mesmo na América Latina podem ajudar no entendimento de tendências populacionais, mas também demandam maior esforço do que o observado em latitudes temperadas, devido à maior riqueza de espécies e diferenças nas dinâmicas de reprodução e migração. Por meio do uso de censos de aves coordenados poderá ser desenvolvida uma técnica para os trópicos que irá gerar informações que permitam acompanhar tendências populacionais, com benefícios para a conservação das aves, similarmente aos censos realizados em países do hemisfério norte.

Bacteriophage- induced reduction in Salmonella Enteritidis counts in the crop of broiler chickens undergoing preslaughter feed withdrawal

Salmonella food poisoning is a public health problem. Feed withdrawal from broiler chickens before slaughter can favor the multiplication of Salmonella in the cecum and crop of contaminated animals and subsequently lead to contamination of carcasses in the processing plant. In the present study, a cocktail of lytic bacteriophages isolated from sewage water was orally administered to 45-d-old broiler chickens 1 h after they received an oral dose of 107 cfu/mL Salmonella enterica subspecies enterica serotype Enteritidis. Immediately after phage administration and 30 min, 1, 3, 6, and 12 h thereafter, groups of chicken were killed. Ceca and crops were analyzed for the presence of Salmonella. At 3 h posttreatment, there were 103 cfu/g and 101 cfu/g of cecal and crop suspension, respectively. At 6 h after treatment, the number of Salmonella was 103 cfu/g in the cecal suspension, but below the detection limit in the crops. our results suggest that bacteriophage therapy may be able to reduce the contamination of chicken carcasses by reducing the preslaughter load of Salmonella in the birds.

Fatores microbiológicos e imunológicos envolvidos no desenvolvimento da cárie precoce da infância

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Characterization of fresh cheese with addition of probiotics and prebiotics

Sensorial and microbiological characteristics of a Brazilian fresh cheese samples with Bifidobacterium animalis subps. lactis as well as samples with this probiotic and polydextrose, a prebiotic ingredient, were evaluated. The addition of this microorganism was studied as: (1) lyophilized probiotic added to cheese curd and (2) by using milk previously fermented by this probiotic to produce the cheese. Cheese samples were microbiologically characterized after 0, 7, 14, 21 and 28 days of storage at a temperature of 4 °C. The microbiological analyses conducted were quantification of total lactic acid bacteria, mesophilic microorganisms, Bif. animalis subps. lactis, coliforms at 30 °C and 45 °C. Affective sensory test was conducted for two different cheese samples (with probiotic and with probiotic and prebiotic) as well as for control one week after manufacturing date. Cheese samples provided acceptable results for coliform counts at 30 °C and 45 °C in compliance with legislation. The cheese samples produced using milk fermented by probiotic showed counts of 107 -108 CFU/g after 28 days of storage, which assures functional property for this product to be claimed.