Endodontic Chelators Induce Nitric Oxide Expression by Murine-cultured Macrophages

Introduction: Endodontic chelators may extrude to apical tissues during instrumentation activating cellular events on periapical tissues. This study assessed in vitro the expression of nitric oxide (NO) concentrations by murine peritoneal macrophages after contact with MTAD (Dentsply/Tulsa, Tulsa, OK), Tetraclean (Ogna Laboratori Farmaceutici, Muggio, Italy), Smear Clear (Sybron Endo, Orange, CA), and EDTA (Biodinamica, Ibipora, PR, Brazil). Methods: Macrophage cells were obtained from Swiss mice after peritoneal lavage. Chelators were diluted in distilled water obtaining 12 concentrations, and MTT assay identified the concentrations, per group, displaying the highest cell viability (analysis of variance, p < 0.01). Selected concentrations were tested for NO expression using Griess reaction. Culture medium and lipopolysaccharide (LPS) were used as controls. Results: Analysis of variance and Tukey tests showed that all chelators displayed elevated NO concentrations compared with the negative control (p < 0.01). MTAD induced the lowest NO expression, followed by Tetraclean, EDTA, and Smear Clear. No difference was observed between MTAD and Tetraclean (p > 0.01), Tetraclean and EDTA (p > 0.01), and EDTA and Smear Clear (p > 0.01). LPS ranked similar to both EDTA and Smear Clear (p > 0.01). Conclusion: The tested endodontic chelators displayed severe proinflammatory effects on murine-cultured macrophages. Citric acid-based solutions induce lower No release than EDTA-based irrigants. (J Endod 2009;35:824-828)

Maximum inhibitory dilution of mouthwashes containing chlorhexidine and polyhexamethylene biguanide against salivary Staphylococcus aureus

Objective: The aim of the present study was to determine the in vitro maximum inhibitory dilution (MID) of two chlorhexidine-based oral mouthwashes (CHX): Noplak (R), Periogard (R), and one polyhexamethylene biguanide-based mouthwash (PHMB): Sanifill Premium (R) against 28 field Staphylococcus aureus strains using the agar dilution method. Materials and Methods: For each product, decimal dilutions ranging from 1/10 to 1/655,360 were prepared in distilled water and added to Mueller Hinton Agar culture medium. After homogenization, the culture medium was poured onto Petri dishes. Strains were inoculated using a Steers multipoint inoculator and dishes were incubated at 37 degrees C for 24hours. For reading, MID was considered as the maximum dilution of the mouthwash still capable of inhibiting microbial growth. Results: Sanifill Premium (R) inhibited the growth of all strains at 1/40 dilution and of 1 strain at 1/80 dilution. Noplak (R) inhibited the growth of 23 strains at 1/640 dilution and of all 28 strains at 1/320 dilution. Periogard (R) showed inhibited growth of 7 strains at 1/640 dilution and of all 28 strains at 1/320 dilution. Data were submitted to Kruskal-Wallis statistical test, showing significant differences between the mouthwashes evaluated (p<0.05). No significant difference was found between Noplak (R) and Periogard (R) (p>0.05). Sanifill Premium (R) was the least effective (p<0.05). Conclusion: It was concluded that CHX-based mouthwashes present better antimicrobial activity against S. Aureus than the PHMB-based mouthwash.

Determination of the maximum inhibitory dilution of cetylpyridinium chloride-based mouthwashes against Staphylococcus aureus: an in vitro study

The aim of this in vitro study was to determine the maximum inhibitory dilution (MID) of four cetylpyridinium chloride (CPC)based mouthwashes: CPC+Propolis, CPC+Malva, CPC+Eucaliptol+Jua+Roma+Propolis (Natural Honey (R)) and CPC (Cepacol (R)), against 28 Staphylococcus aureus field strains, using the agar dilution method. Decimal dilutions ranging from 1/10 to 1/655,360 were prepared and added to Mueller Hinton Agar. Strains were inoculated using Steers multipoint inoculator. The inocula were seeded onto the surface of the culture medium in Petri dishes containing different dilutions of the mouthwashes. The dishes were incubated at 37 degrees C for 24 h. For readings, the MID was considered as the maximum dilution of mouthwash still capable of inhibiting microbial growth. The obtained data showed that CPC+Propolis had antimicrobial activity against 27 strains at 1/320 dilution and against all 28 strains at 1/160 dilution, CPC+Malva inhibited the growth of all 28 strains at 1/320 dilution, CPC+Eucaliptol+Jua+Roma+Propolis inhibited the growth of 2 strains at 1/640 dilution and all 28 strains at 1/320 dilution, and Cepacol (R) showed antimicrobial activity against 3 strains at 1/320 dilution and against all 28 strains at 1/160 dilution. Data were submitted to Kruskal-Wallis test, showing that the MID of Cepacol (R) was lower than that determined for the other products (p<0.05). In conclusion, CPC-mouthwashes showed antimicrobial activity against S. aureus and the addition of other substances to CPC improved its antimicrobial effect.

Atividade antifúngica de óleos essenciais frente a amostras clínicas de Candida albicans isoladas de pacientes HIV positivos

Objetivou-se avaliar a atividade antifúngica dos óleos essenciais de Ocimum basilicum L. (manjericão), Cymbopogon martinii L. (palmarosa), Thymus vulgaris L. (tomilho) e Cinnamomum cassia Blume (canela da china) sobre cepas de Candida albicans isoladas de pacientes HIV positivos e cepa padrão (ATCC 76845). Quinze amostras clínicas de C. albicans (C1-C15) foram repicadas em ágar Sabouraud Dextrose, para confecção de suspensões em solução salina estéril (0,9%) contendo 1,5 x 10(6) UFC mL-1. As emulsões dos óleos essenciais foram preparadas em água destilada estéril e tween 80, com concentrações variando entre 1024 µg mL-1 e 4 µg mL-1. A ação antifúngica foi determinada por meio da Concentração Inibitória Mínima (CIM) utilizando-se a técnica da microdiluição. Foram utilizados como controles positivos a nistatina e o miconazol (50 µg mL-1). Os testes foram realizados em triplicata, sendo a CIM, a menor concentração capaz de inibir o crescimento das leveduras, observada por método visual de acordo com a turvação do meio de cultura. Para C. albicans (ATCC 76845), a CIM do óleo essencial de C. cassia foi 64 µg mL-1, enquanto para óleo de C. martinii foi 1024 µg mL-1. Para as cepas clínicas, verificou-se que a CIM de C. cassia para 80% das cepas foi 64 µg mL-1, sendo a variação dos valores da CIM entre 128 µg mL-1 e 64 µg mL-1. Observou-se que para 66,6% das amostras clínicas, a CIM de C. martinii foi 612 µg mL-1. Constatou-se que a nistatina não apresentou atividade frente às cepas clínicas (C1-C15), enquanto a atividade antifúngica do miconazol foi verificada em 100% das amostras. Não se constatou atividade antimicrobiana dos óleos essenciais de O. basilicum e T. vulgaris, nas concentrações avaliadas. Concluiu-se que os óleos essenciais de C. cassia e C. martinii, em diferentes concentrações, apresentam atividade antifúngica sobre cepas de C. albicans isoladas de pacientes HIV positivos e cepa padrão (ATCC 76845). Entretanto não foi observada inibição antimicrobiana para os óleos de O. basilicum e T. vulgaris.

The fibroblast growth factor family: involvement in the regulation of folliculogenesis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ecological study of Paracoccidioides brasiliensis in soil: growth ability, conidia production and molecular detection

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

Production of reactive oxygen (H2O2) and nitrogen (NO) intermediates and TNF-α in mice genetically selected for high (H) and low (L) antibody response and experimentally infected with Leptospira serovar pomona

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Atividade antibacteriana de óleos essenciais de plantas frente a linhagens de Staphylococcus aureus e Escherichia coli isoladas de casos clínicos humanos

A ação antibacteriana in vitro de óleos essenciais de seis plantas foi verificada por meio da Concentração Inibitória Mínima (CIM=%v/v) pela diluição dos óleos em meio de cultura Mueller Hinton Agar, frente a linhagens de Staphylococcus aureus (n=16) e Escherichia coli (n=16) isoladas de casos clínicos humanos, além de 1 amostra padrão ATCC para cada espécie (Sa ATCC 25923 e Ec ATCC 25922), e determinação de curvas de sobrevivência em concentrações equivalentes a CIM90% dos respectivos óleos. O óleo essencial de canela foi o mais eficiente, com valores de CIM90% de 0,047 e 0,09 para S. aureus e E. coli respectivamente, enquanto gengibre (0,09), cravo da índia (0,095) e capim cidreira (0,1) apresentaram eficiências semelhantes para S. aureus. Frente a E. coli, os óleos de gengibre (0,52) e capim cidreira (0,55) foram equivalentes quanto à eficiência. de acordo com as curvas de sobrevivência, foi possível verificar também que os valores de CIM90% obtidos podem ser tanto bactericidas ou bacteriostáticas de acordo com a bactéria testada. em conclusão, verificou-se que os óleos essenciais testados foram efetivos no controle do desenvolvimento bacteriano, sendo o potencial antimicrobiano diferente em função da espécie bacteriana testada, sendo que a bactéria Gram positiva (S. aureus) mostrou-se mais susceptível aos óleos testados que a Gram negativa (E. coli).

Efficacy of the nematode-trapping fungus Duddingtonia flagrans against infections by Haemonchus and Trichostrongylus species in lambs at pasture

The efficacy of the nematode-trapping fungus Duddingtonia flagrans against infections by trichostrongyle nematodes in sheep was assessed throughout 6 months. Twenty Ile de France lambs were divided into two groups (control and treated groups), which were kept in separate pastures. Animals of the treated group were fed with D. flagrans twice a week (Tuesdays and Fridays). Pellets were prepared with the fungus mycelia in liquid culture medium and contained approximately 20% fungus. They were mixed with the animals' diet at a concentration of 1 g pellet per 10 kg live weight. Faecal egg counts (FEC), packed cell volume (PCV), total serum protein and the animals' body weight were determined fortnightly from 7 October 2005 to 24 March 2006. Comparison of such parameters between groups showed no significant differences (P > 0.05), except on 10 February 2006, when the control group presented a higher mean FEC than the treated group (P < 0.05). Feeding sheep with pellets containing D. flagrans had no benefit to the prophylaxis of nematode infections under the experimental conditions used in the present study.

Endogenous and exogenous polyamines in the organogenesis in Curcuma longa L.

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

Optimization of cyclodextrin glucanotransferase production from Bacillus clausii E16 in submerged fermentation using response surface methodology

Cyclodextrin glucanotransferase production from Bacillus clausii E16, a new bacteria isolated from Brazilian soil samples was optimized in shake-flask cultures. A 2 4 full-factorial central composite design was performed to optimize the culture conditions, using a response surface methodology the combined effect among the soluble starch concentration, the peptone concentration, the yeast extract concentration, and the initial pH value of the culture medium was investigated. The optimum concentrations of the components, determined by a 2(4) full-factorial central composite design, were 13.4 g/L soluble starch, 4.9 g/L peptone, 5.9 g/L yeast extract, and initial pH 10.1. Under these optimized conditions, the maximum cyclodextrin glucanotransferase activity was 5.9 U/mL after a 48-h fermentation. This yield was 68% higher than that obtained when the microorganism was cultivated in basal culture medium.

Production of extracellular acid proteases by Aspergillus clavatus

The production of extracellular acid proteases from Aspergillus clavatus was evaluated in a culture filtrate medium, with different carbon and nitrogen sources. The fungus was cultivated at three different temperatures during 10 days. The proteolytic activity was determined on haemoglobin pH 5.0 at 37 degreesC. The highest acid proteolytic activity (80 U/ml) was observed in culture medium containing glucose and gelatin at 1% (w/v) at 30 degreesC at the third day of incubation. Cultures developed in Vogel medium with glucose at 2% (w/v) showed at about 45% of proteolytic activity when compared to the cultures with 1% of the same sugar. The optimum pH of enzymatic activity was 2.0 and the enzyme was stable at pH values ranging from 2.0 to 4.0. The optimum temperature was 40 degreesC and the half-lives at 40, 45 and 50 degreesC were 30, 10 and 5 min, respectively.

Purification and physicochemical properties of a ribonuclease produced by Aspergillus flavipes (IZ : 1501)

A RNase of Aspergillus flavipes (IZ:1501) was purified from culture medium by chromatography on DEAE-cellulose and Sephadex G50 columns, after 96 h of cultivation. The molecular weight of the RNase was estimated to be 15 kD by gel filtration using Sephadex G100, and the optimum pH and temperature were 4.0 and 55 degrees C, respectively. Catalytic activity was inhibited by Hg2+, Ag+, Fe3+, Co2+ and Zn2+. The enzyme showed guanosine specificity producing only 3'-GMP from yeast RNA.

Bacillus lehensis-an alkali-tolerant bacterium isolated from cassava starch wastewater: optimization of parameters for cyclodextrin glycosyltransferase production

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Production of xylanolytic enzymes by Penicillium janczewskii

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)