Tempol decreases the levels of reactive oxygen species in human neutrophils and impairs their response to

SUMMARY Background and aims: Mycobacterium tuberculosis (Mtb), the main causative agent of human tuberculosis (TB), remains as a serious public health problem. The innate immune response following Mtb infection plays a crucial role in preventing the onset of active TB and limiting its spread. Since phagocytes-derived reactive oxygen/nitrogen species (ROS/RNS) during the oxidative burst can fight Mtb, we hypothesized that the use of antioxidants could increase the host's susceptibility to Mtb/TB. In that way, we investigated the effects of the nitroxide Tempol, an antioxidant with superoxide dismutase-like activity, on the response of neutrophils against Mtb. Methods: Human blood-derived neutrophils were isolated from healthy volunteers and incubated with Mtb (H37Ra) at different multiplicities of infection (MOIs), in the absence or presence of Tempol. The levels of ROS in neutrophils were evaluated using the cytochrome C reduction assay (extracellular O2 •-) and luminol-(total intracellular and extracellular ROS) and isoluminol-(extracellular ROS) amplified chemiluminescence assay. The killing assay (two-step protocol) checked the mycobactericidal capacity of neutrophils, as calculated the phagocytosis (K p ) and intracellular killing (Kk) rates. Results: The levels of ROS and killing capacity in Mtb-stimulated neutrophils were significantly decreased by 450 µM Tempol (p < 0.05). Interestingly, Tempol decreased the k k of neutrophils, but did not affect their kp, demonstrating that a putative diminution in ROS levels, ultimately, affected the intracellular killing of Mtb. Conclusion: This study provides insights regarding the role of antioxidants on the neutrophil response toward Mtb, so that our findings deserve to be considered regarding further studies and clinical implications.

Contextualización y objetivos: Mycobacterium tuberculosis (Mtb), el principal agente causal de la tuberculosis humana (TB), es un grave problema de salud pública. La respuesta inmune innata desencadenada en la infección por Mtb juega un papel crucial en la prevención de la aparición de la tuberculosis activa y en la limitación de su propagación. Dado que las especies reactivas de oxígeno/nitrógeno derivadas de los fagocitos (ERO/ERN) durante el burst oxidativo pueden combatir la infección pulmonar por Mtb, planteamos la hipótesis de que el uso de antioxidantes podría aumentar la susceptibilidad del huésped humano hacia Mtb/TB. De esa manera, investigamos los efectos del nitróxido Tempol, un antioxidante con actividad similar a la superóxido dismutasa, sobre la respuesta de los neutrófilos contra Mtb. Métodos: se aislaron neutrófilos derivados de sangre humana de voluntarios sanos y se incubaron con Mtb (H37Ra) en diferentes multiplicidades de infección (MOI), en ausencia o presencia de Tempol. Los niveles de ERO en neutrófilos se evaluaron mediante el ensayo de la reducción del citocromo C (O2'- extracelular) y el ensayo de quimioluminiscencia, amplificada mediante el uso de luminol (ERO total, intracelular y extracelular) e isoluminol (ERO extracelular). La prueba de actividad microbicida (protocolo de dos pasos) verificó la capacidad micobactericida de los neutrófilos, calculando las tasas de fagocitosis (Kp) y muerte intracelular (Kk). Resultados: los niveles de ERO y la capacidad micobactericida en neutrófilos estimulados con Mtb disminuyeron significativamente en los grupos tratados con 450 µM de Tempol (p < 0,05). Curiosamente, Tempol disminuyó la tasa de muerte intracelular (Kk) en neutrófilos, pero no tuvo ningún efecto sobre la tasa de fagocitosis (Kp), lo que demuestra que una supuesta disminución en los niveles de ERO, en última instancia, afectó la destrucción intracelular de Mtb. Conclusión: este estudio proporciona información sobre el papel de los antioxidantes en la respuesta de los neutrófilos hacia Mtb, por lo que nuestros hallazgos merecen ser considerados con respecto a más estudios e implicaciones clínicas.

Contextualização e objetivos: Mycobacterium tuberculosis (Mtb), principal agente causal da tuberculose humana (TB), é um grave problema de saúde pública. A resposta imune inata desencadeada pela infecção por Mtb desempenha um papel crucial na prevenção do aparecimento da tuberculose ativa e na limitação de sua disseminação. Como as espécies reativas de oxigênio/nitrogênio derivadas de fagó-citos (ERO/ERN) durante a burst oxidativa podem combater a infecção pulmonar por Mtb, hipotetizamos que o uso de antioxidantes poderia aumentar a suscetibilidade do hospedeiro humano ao Mtb/TB. Assim, investigamos os efeitos do nitróxido de Tempol, um antioxidante com atividade semelhante a superóxido dismutase, na resposta de neutrófilos contra o Mtb. Métodos: neutrófilos derivados de sangue humano foram isolados de voluntários saudáveis e incubados com Mtb(H37Ra) em diferentes multiplicidades de infecção (MOI), na ausência ou presença de Tempol. Os níveis de ERO em neutrófilos foram avaliados por ensaio de depleção de citocromo C (O2•- extracelular) e ensaio de quimioluminescência, amplificado com luminol (ERO total, intracelular e extracelular) e isoluminol (ERO extracelular). O teste de atividade microbicida (protocolo de duas etapas) verificou a capacidade micobactericida dos neutrófilos, calculando as taxas de fagocitose (Kp) e morte intracelular (Kk). Resultados: os níveis de ERO e a capacidade micobactericida em neutrófilos estimulados por Mtb diminuíram significativamente nos grupos tratados com Tempol 450 µM (p < 0,05). Curiosamente, Tempol diminuiu a taxa de morte intracelular (Kk) em neutrófilos, mas não teve efeito sobre a taxa de fagocitose (Kp), mostrando que uma diminuição putativa nos níveis de ERO afetou a morte intracelular de Mtb. Conclusão: este estudo fornece informações sobre o papel dos antioxidantes na resposta dos neutrófilos ao Mtb, portanto, nossos achados merecem consideração para estudos adicionais e implicações clínicas.

Photobiomodulation enhances the Th1 immune response of human monocytes.

This study aims to evaluate the effects of photobiomodulation (PBM) on human monocytes, assessing the oxidative burst and ultimate fungicidal potential of these cells, as well as the gene expression at the mRNA level of CD68, CD80, CD163, CD204, IL-6, TNF-α and IL-10 in derived macrophages. Primary cultures of human monocytes were irradiated with an InGaAlP (660 nm)/GaAlAs (780 nm) diode laser (parameters: 40 mW, 0.04 cm2, 1 W/cm2; doses: 200, 400 and 600 J/cm2). Cells were submitted to the chemiluminescence assay, and a microbicidal activity assay against Candida albicans was performed. Reactive oxygen species (ROS) and nitric oxide (NO) production were measured, and cell viability was assessed by the exclusion method using 0.2% Trypan blue reagent. Irradiated monocytes were cultured for 72 h towards differentiation into macrophages. Total RNA was extracted, submitted to reverse transcription and real-time PCR. The results were analysed by ANOVA and the Tukey test (α = 0.05). Irradiated monocytes revealed a significant increase in their intracellular and extracellular ROS (P < 0.001). The 660 nm wavelength and 400 J/cm2 dose were the most relevant parameters (P < 0.001). The fungicidal capacity of the monocytes was shown to be greatly increased after PBM (P < 0.001). PBM increased the expression of TNF-α (P = 0.0302) and the production of NO (P < 0.05) and did not impair monocyte viability. PBM induces a pro-inflammatory Th1-driven response in monocytes and macrophages.

Seroepidemiological survey on sporotrichosis-infection in rural areas of the south of Minas Gerais State, Brazil.

Sporotrichosis is a subcutaneous mycosis caused by traumatic inoculation into the skin by fungi species of the genus Sporothrix. The disease has different clinical manifestations (cutaneous, lymphocutaneous, and disseminated), and can also progress to a systemic infection. Despite having a worldwide distribution, sporotrichosis is most prevalent in tropical and subtropical countries. In Brazil, reports of the disease are higher frequent, where cases of the disease were found in Rio de Janeiro, Sao Paulo, Curitiba, Pernambuco, and Paraiba, among others. Certain groups of people may be more exposed to the causative agent of disease, such as residents of rural areas. Thus, this work aimed to carry out a seroepidemiological survey of the prevalence of sporotrichosis in four rural locations in the south of Minas Gerais State, Brazil. In this study, we used an indirect ELISA test in the survey on the prevalence of sporotrichosis. Data obtained in this study evaluated a population of 631 individuals and showed a prevalence of 44.69%. The distribution of seroprevalence of sporotrichosis with respect to age groups and gender showed no significant statistical difference. Thus, we found a high seroprevalence of sporotrichosis-infection in rural regions of southern Minas Gerais State, Brazil, with no difference in prevalence in relation to gender and age.

Detection of Serum Antibodies and Peripheral Blood Mononuclear Cells Response to Escherichia coli Antigens in Humans

Abstract Among the microorganisms that make up the intestinal microbiota, stands out Escherichia coli, which has as main ecological niche, the large human intestine. Its importance stands out in being part of the pioneer's commensal microorganisms on the colonization of the intestinal mucosa and its pathogenic role causing extra and intra intestinal diseases. The aim of the study was to evaluate the antibody production and proliferative response of Peripheral Blood Mononuclear Cells (PBMC) to E. coli antigens. The bacteria were grown on Brain Heart Infusion broth medium at 35 ºC for 72 hours. Pellet bacteria were lysed for one hour at room temperature with an 8M sodium guanidine solution. After spin and dialysis, the protein antigens were measured in the supernatant by protein assay. The antigens were characterized by polyacrylamide gel electrophoresis and the antigenic profile by western blotting. The presence of specific IgG and IgA antibodies were evaluated using thirty normal human sera by an indirect ELISA. The response of PBMC to E. coli antigens was assessed by MTT metabolization. The results demonstrated that the antigens were composed of proteins of different sizes and they were recognized by antibodies present in normal human serum. Human sera presented high titers of IgG and IgA antibodies to E. coli antigens when compared to the results of lipopolysaccharide. We also showed that total E. coli antigens induced PBMC proliferation at different antigen concentrations. Taken together the results suggest that the antigens from E. coli can induce local and systemic immune responses.

Study and evaluation of antimicrobial activity and antioxidant capacity of dry extract and fractions of leaves of Raphanus sativus var. oleiferus Metzg / Estudo e avaliação da atividade antimicrobiana e capacidade antioxidante do extrato seco e frações de folhas de Raphanus sativus var. oleiferus Metzg

The radish (Raphanus sativus L.) is a vegetable of the Brassicaceae family cultivated worldwide and has several medicinal properties. Its biological activities are related to various secondary metabolites present in the species, especially phenolics. Thus, the objectives of this study were the chemical analysis and evaluation of the antioxidant and antimicrobial activities of the dry extract and fractions of the fodder turnip leaves (R. sativus var. oleiferus Metzg.). Samples were analyzed by mass spectrometry and the antioxidant activity was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical method and the reducing power method. Antimicrobial activity was determined by the agar diffusion and microdilution methods. The total phenols were concentrated in the butanol fraction (121.27 mg GAE/g) and the flavonoids were concentrated in the ethyl acetate fraction (98.02 mg EQ/g). The ethyl acetate fraction showed the best antioxidants results, with 83.45% of free radical scavenging and 11.34% of ferric ions reduction. The analysis of antimicrobial activity showed that the dry extract had the highest average zone of inhibition against Bacillus subtilis (18.67 mm). Smaller values of the minimum inhibitory concentration for Micrococcus luteus were, and the ethyl acetate fraction showed a lower minimum inhibitory concentration (0.1 mg/ml) for that microorganism. There was a strong correlation between the antioxidant activity and the content of phenols and flavonoids. The results showed the potential antioxidant and antimicrobial activities of this extract with the ethyl acetate fraction being most promising for further studies.

O rabanete(Raphanus sativus L.) é um vegetal da família Brassicaceae cultivado em todo o mundo e possui diversas propriedades medicinais. Suas atividades biológicas estão relacionadas aos vários metabólitos secundários presentes na espécie, especialmente os compostos fenólicos. Desta forma, os objetivos deste estudo foram realizar análises químicas e avaliar as atividades antioxidante e antimicrobiana do extrato seco e das frações das folhas de R. sativus var. oleiferus Metzg. As amostras foram analisadas em espectrômetro de massas e o potencial antioxidante foi avaliado pelos métodos do radical DPPH (2,2-difenil-1-picrilhidrazila) e do poder redutor. A atividade antimicrobiana foi determinada pelos métodos de difusão em ágar e da microdiluição. Observou-se que os fenóis totais se concentraram na fração butanólica (121,27 mg EAG/g), enquanto que e os teores de flavonoides concentraram-se na fração acetato de etila (98,02 mg EQ/g). A fração acetato de etila apresentou os melhores resultados antioxidantes, com porcentagem de sequestro dos radicais DPPH de 83,45% e com porcentagem de redução dos íons férrico de 11,34%. A análise da atividade antimicrobiana revelou que o extrato seco teve maior média de halos de inibição frente ao Bacillus subtilis(18,67 mm). Os menores valores da concentração inibitória mínima foram para Micrococcus luteus, sendo que a fração acetato de etila demonstrou menor concentração inibitória mínima (0,1 mg/mL) para esse micro-organismo. Houve uma forte correlação entre a atividade antioxidante e o teor de fenóis e de flavonoides. Os resultados demonstraram potenciais ações antioxidante e antimicrobiana do extrato e das frações avaliados, sendo a fração acetato de etila promissora para estudos posteriores.

Multicenter study of Candida species in oral mucosa of different patients: analysis of 711 strains and literature review

This study aimed to identify Candida spp. collected from oral mucosa and maintained in culture collections, correlating the findings with the medical history of patients and comparing with data from the literature over the past five years. Seven hundred and eleven oral Candida spp. isolates, collected between 2013 and 2017, were selected and identified using traditional and molecular methods. In addition, a literature review was performed with the key words: "Oral", "Candida" and "Yeast". Seven species of the genus Candida: were identified C. albicans(73.3%); C. tropicalis (9.3%); C. parapsilosis (8.2%); C. glabrata (3.9%); C. guilliermondii(2.8%); C. krusei (1.7%) and C. lusitaniae (0.3%). The strains identified as C. albicans were submitted to molecular methods using specific primers and of these, 5.8% were identified as C. dubliniensis strains. The greatest diversity of strains was found in patients presenting no systemic diseases or HIV +, while the highest percentage of strains of Candidanon-albicanswere observed in cancer patients. This study reports a representative distribution of Candidaspecies among individuals exhibiting distinct clinical conditions, in order to contribute to the design of future research on details of aspects involved in the infections caused by these microorganisms. The correct identification of oral Candida strains contributes to a realistic epidemiological approach and future clinical protocols against these pathogens

Correction to: Effects of metaperiodate and urea solutions on the serological diagnosis of human sporotrichosis using an indirect ELISA test.

In the article mentioned above an author's name was misspelled. The correct author name reads as follows: Leila Maria Lopes-Bezerra. We apologize for the inconvenience.

Effects of metaperiodate and urea solutions on the serological diagnosis of human sporotrichosis using an indirect ELISA test.

Sporotrichosis is an infection of the skin caused by traumatic inoculation of the fungus Sporothrix schenckii. Definitive diagnosis relies on direct visualization of the fungus or its isolation on culture medium, although both have low sensitivity. Alternatively, the detection of the antibody response offers a more rapid alternative for diagnosis. Although the available immunoassays possess good sensitivity and specificity, cross-reactivity is still a problem. This study aimed to evaluate the effect of sodium metaperiodate and 6 M urea solutions on the serological diagnosis of sporotrichosis using an enzyme-linked immunosorbent assay (ELISA) test. Ninety-six-well plates were sensitized with exoantigens from the yeast phase of S. schenckii. Sera of patients with confirmed sporotrichosis, sera of patients with paracoccidioidomycosis, and sera of individuals with a sporotrichin-negative skin test were tested. Two strategies were used; the first consisted of treating the antigen with sodium metaperiodate solution for different incubation times, and the second consisted of treating the serum with 6 M urea solution for different incubation times. ROC curve analysis revealed that the best discrimination parameters were obtained using 6 M urea solution incubated for 5 min and serum dilution at 1/600. The use of 6 M urea solution improves the performance of the ELISA test in the diagnosis of sporotrichosis.

Antimicrobial activity of various brands of children's toothpastes formulated with Triclosan, fluoride and xylitol / Atividade antimicrobiana de dentifrícios infantis formulados com Triclosa, xylitol e fluoreto, disponíveis no mercado

Objective: Evaluate and compare the antimicrobial efficacy of four different commercially available children's toothpastes on bacteria growth inhibition from oral microbiota of young children. Material and Method: The toothpastes tested contained: 1100 ppm sodium fluoride, xylitol and 750 ppm sodium fluoride, xylitol and 500 ppm sodium fluoride, and xylitol and triclosan. Twenty 18 to 36 month-old children were selected. Nonstimulated saliva samples were collected and inoculated on brain heart infusion (BHI) agar. Toothpaste dilutions were applied into the agar wells and incubated at 37 ºC for 24 hours in microaerophilic conditions. Saline and 0.12% chlorhexidine were used as controls. Antimicrobial activity was determined in duplicate by agar-well diffusion technique. Bacterial growth inhibition zones were recorded in mm. Statistical analysis was performed using ANOVA and Tukey's test (p < 0.05). Results: All toothpastes showed antimicrobial activity when compared to negative control (p < 0.05). Toothpaste containing triclosan presented the highest antimicrobial activity followed, in this order, by xylitol and 750 ppm sodium fluoride, chlorhexidine, 1100 ppm sodium fluoride, xylitol and 500 ppm sodium fluoride and saline. Conclusion: Toothpaste containing triclosan and xylitol presented excellent antimicrobial activity, and may be considered a good option for young children. (AU)

Objetivo: Avaliar e comparar a atividade antimicrobiana de 4 dentifrícios infantis, comercialmente disponíveis, com relação à inibição do crescimento da microbiota oral em crianças. Material e Métodos: Os dentifrícios testados continham: 1100 ppm de fluoreto de sódio - xilitol e 750 ppm de fluoreto de sódio - xilitol e 500 ppm de fluoreto de sódio - xilitol e triclosan. Vinte crianças de 18 a 36 meses participaram dessa pesquisa. Amostras de saliva não estimulada foram coletadas e inoculadas em ágar de infusão cérebro-coração (ICC). Dentifrícios diluídos foram aplicados sobre os poços contendo ICC e incubados a 37 ºC por 24 horas em condições microaerofílicas. Solução salina e clorexidina a 0,12% foram usadas como controles. A atividade antimicrobiana foi determinada em duplicata pela técnica de difusão nos poços de ágar. Os halos de inibição do crescimento bacteriano foram registrados em mm. A análise estatística foi realizada usando ANOVA e teste de Tukey (p < 0,05). Resultados: Todos os dentifrícios mostraram atividade antimicrobiana, quando comparados com os controles negativos. O dentifrício contendo triclosan mostrou a maior atividade antimicrobiana, seguido, nessa ordem por dentifrício contendo xilitol e 750 ppm de fluoreto de sódio, clorexidina, dentifrício contendo 1100 ppm de fluoreto de sódio, dentifrício contendo xilitol e 500 ppm de fluoreto de sódio e pela solução salina. Conclusão: O dentifrício contendo triclosan e xylitol apresentou excelente atividade antimicrobiana podendo ser considerado uma vantajosa opção para crianças, contribuindo para o controle das cáries dentárias e reduzindo a ingestão de fluoretos.(AU)

Genotoxicidade de Helianthus annuus Linné, 1753 (Asteraceae): novas perspectivas e tendências / Genotoxicity of Helianthus annuus Linné, 1753 (Asteraceae): new perspectives and trends

A presente revisão reúne informações acerca dos aspectos genotóxicos de Helianthus annuus Linné (girassol), até o presente momento na literatura. O girassol é uma importante fonte de óleo natural e sua ampla aplicabilidade é atribuível ao variável repertório fitoquímico. A ação preventiva da diurese, diarreia e doenças inflamatórias, além dos efeitos de alívio dos sintomas asmáticos, proteção gástrica, cicatrização, ação anti-inflamatória e antimicrobiana foram reconhecidas quanto às propriedades farmacológicas do girassol. Com um grande potencial fitoquímico, é importante também analisarmos seu potencial tóxico e genotóxico. Vários resultados inconclusivos a respeito da correlação entre a carcinogênese e o óleo de girassol foram observados na literatura. Por outro lado, um número limitado de informações sobre a mutagênese ou antimutagênese do extrato hidroalcoólico e distintas fontes do óleo de sementes de girassol, submetido ou não ao estresse térmico, foi relatado. Esta revisão apresentará os atuais trabalhos envolvendo a ação genotóxica ou anti genotóxica de H. annuus L., colaborando com a implantação de limites ao consumo, potenciais riscos à saúde ou medidas estratégicas quimiopreventivas.(AU)

The present review gathers information about the genotoxic aspects of Helianthus annuus Linné (sunflower), until the present moment in the literature. Sunflower is an important source of natural oil and its wide applicability is attributable to the variable phytochemical repertoire. The preventive action of diuresis, diarrhea and inflammatory diseases, besides the effects of relief of asthmatic symptoms, gastric protection, healing, anti-inflammatory and antimicrobial action were recognized regarding the pharmacological properties of the sunflower. With great phytochemical potential, it is also important to analyze its toxic and genotoxic potential. Several inconclusive results regarding the correlation between carcinogenesis and sunflower oil were observed in the literature. On the other hand, a limited number of information on the mutagenesis or anti-mutagenesis of the hydroalcoholic extract and different sources of sunflower seed oil, whether or not subjected to thermal stress, was reported. This review will present the current works involving the genotoxic or anti genotoxic action of H. annuus L., collaborating with the implementation of consumption limits, potential health risks or strategic chemopreventive measures.(AU)

Low-level laser therapy to the mouse femur enhances the fungicidal response of neutrophils against Paracoccidioides brasiliensis.

Neutrophils (PMN) play a central role in host defense against the neglected fungal infection paracoccidioidomycosis (PCM), which is caused by the dimorphic fungus Paracoccidioides brasiliensis (Pb). PCM is of major importance, especially in Latin America, and its treatment relies on the use of antifungal drugs. However, the course of treatment is lengthy, leading to side effects and even development of fungal resistance. The goal of the study was to use low-level laser therapy (LLLT) to stimulate PMN to fight Pb in vivo. Swiss mice with subcutaneous air pouches were inoculated with a virulent strain of Pb or fungal cell wall components (Zymosan), and then received LLLT (780 nm; 50 mW; 12.5 J/cm2; 30 seconds per point, giving a total energy of 0.5 J per point) on alternate days at two points on each hind leg. The aim was to reach the bone marrow in the femur with light. Non-irradiated animals were used as controls. The number and viability of the PMN that migrated to the inoculation site was assessed, as well as their ability to synthesize proteins, produce reactive oxygen species (ROS) and their fungicidal activity. The highly pure PMN populations obtained after 10 days of infection were also subsequently cultured in the presence of Pb for trials of protein production, evaluation of mitochondrial activity, ROS production and quantification of viable fungi growth. PMN from mice that received LLLT were more active metabolically, had higher fungicidal activity against Pb in vivo and also in vitro. The kinetics of neutrophil protein production also correlated with a more activated state. LLLT may be a safe and non-invasive approach to deal with PCM infection.

In vitro screening antibacterial activity of Bidens pilosa Linné and Annona crassiflora Mart. against oxacillin resistant Staphylococcus aureus (ORSA) from the aerial environment at the dental clinic.

Currently multiresistant Staphylococcus aureus is one common cause of infections with high rates of morbidity and mortality worldwide, which directs scientific endeavors in search for novel antimicrobials. In this study, nine extracts from Bidens pilosa (root, stem, flower and leaves) and Annona crassiflora (rind fruit, stem, leaves, seed and pulp) were obtained with ethanol: water (7:3, v/v) and their in vitro antibacterial activity evaluated through both the agar diffusion and broth microdilution methods against 60 Oxacillin Resistant S. aureus (ORSA) strains and against S. aureus ATCC6538. The extracts from B. pilosa and A. crassiflora inhibited the growth of the ORSA isolates in both methods. Leaves of B. pilosa presented mean of the inhibition zone diameters significantly higher than chlorexidine 0.12% against ORSA, and the extracts were more active against S. aureus ATCC (p < 0.05). Parallel, toxicity testing by using MTT method and phytochemical screening were assessed, and three extracts (B. pilosa, root and leaf, and A. crassiflora, seed) did not evidence toxicity. On the other hand, the cytotoxic concentrations (CC50 and CC90) for other extracts ranged from 2.06 to 10.77 mg/mL. The presence of variable alkaloids, flavonoids, tannins and saponins was observed, even though there was a total absence of anthraquinones. Thus, the extracts from the leaves of B. pilosa revealed good anti-ORSA activity and did not exhibit toxicity.

In vitro SCREENING ANTIBACTERIAL ACTIVITY OF Bidens pilosa LINNÉ AND Annona crassiflora MART. AGAINST OXACILLIN RESISTANT Staphylococcus aureus (ORSA) FROM THE AERIAL ENVIRONMENT AT THE DENTAL CLINIC / Triagem in vitro da atividade antibacteriana de Bidens pilosa Linné e Annona crassiflora Mart. contra Staphylococcus aureus resistente à oxacilina (ORSA) provenientes do ambiente aéreo na clínica odontológica

Currently multiresistant Staphylococcus aureus is one common cause of infections with high rates of morbidity and mortality worldwide, which directs scientific endeavors in search for novel antimicrobials. In this study, nine extracts from Bidens pilosa (root, stem, flower and leaves) and Annona crassiflora (rind fruit, stem, leaves, seed and pulp) were obtained with ethanol: water (7:3, v/v) and their in vitro antibacterial activity evaluated through both the agar diffusion and broth microdilution methods against 60 Oxacillin Resistant S. aureus (ORSA) strains and against S. aureus ATCC6538. The extracts from B. pilosa and A. crassiflora inhibited the growth of the ORSA isolates in both methods. Leaves of B. pilosa presented mean of the inhibition zone diameters significantly higher than chlorexidine 0.12% against ORSA, and the extracts were more active against S. aureus ATCC (p < 0.05). Parallel, toxicity testing by using MTT method and phytochemical screening were assessed, and three extracts (B. pilosa, root and leaf, and A. crassiflora, seed) did not evidence toxicity. On the other hand, the cytotoxic concentrations (CC50 and CC90) for other extracts ranged from 2.06 to 10.77 mg/mL. The presence of variable alkaloids, flavonoids, tannins and saponins was observed, even though there was a total absence of anthraquinones. Thus, the extracts from the leaves of B. pilosa revealed good anti-ORSA activity and did not exhibit toxicity.

Atualmente Staphylococcus aureus multirresistente é causa comum de infecções com altas taxas de morbidade e mortalidade mundialmente, o que direciona esforços científicos na busca de novos antimicrobianos. Neste estudo, nove extratos de Bidens pilosa (raiz, caule, flor e folhas) e de Annona crassiflora (casca do fruto, caule, folha, semente e polpa) foram obtidos com etanol:água (7:3, v/v) e suas atividades antibacteriana in vitro avaliadas através de difusão em agar e microdiluição em caldo contra 60 cepas de Oxacillin Resistant S. aureus (ORSA) e contra S. aureus ATCC 6538. Os extratos de B. pilosa e A. crassiflora inibiram o crescimento dos isolados ORSA em ambos os métodos. O extrato da folha de B. pilosa apresentou média dos diâmetros dos halos de inibição significativamente maior que a clorexidina 0,12%, contra os isolados ORSA, e os extratos foram mais ativos contra S. aureus ATCC (p < 0,05). Paralelamente, teste de toxicidade pelo método MTT e triagem fitoquímica foram avaliadas, e três extratos (raiz e folha de B. pilosa e semente de A. crassiflora) não apresentaram toxicidade. Por outro lado, as concentrações citotóxicas (CC50 e CC90) para os outros extratos variaram de 2,06 a 10,77 mg/mL. Observou-se variável presença de alcalóides, flavonóides, taninos e saponinas, apesar de total ausência de antraquinonas. Portanto, os extratos das folhas de B. pilosa revelaram boa atividade anti-ORSA e não exibiram toxicidade.

Evaluation of antimicrobial and cytotoxic activities of plant extracts from southern Minas Gerais cerrado.

The antimicrobial activity of plant hidroethanolic extracts on bacteria Gram positive, Gram negative, yeasts, Mycobacterium tuberculosis H37 and Mycobacterium bovis was evaluated by using the technique of Agar diffusion and microdilution in broth. Among the extracts evaluated by Agar diffusion, the extract of Bidens pilosa leaf presented the most expressive average of haloes of growth inhibition to the microorganisms, followed by the extract of B. pilosa flower, of Eugenia pyriformis' leaf and seed, of Plinia cauliflora leaf which statistically presented the same average of haloes inhibitory formation on bacteria Gram positive, Gram negative and yeasts. The extracts of Heliconia rostrata did not present activity. Mycobacterium tuberculosis H37 and Mycobacterium bovis (BCG) appeared resistant to all the extracts. The susceptibility profile of Candida albicans and Saccharomyces cerevisiae fungi were compared to one another and to the Gram positive Bacillus subtilis, Enterococcus faecalis and the Gram negative Salmonella typhimurium bacteria (p > 0.05). The evaluation of cytotoxicity was carried out on C6-36 larvae cells of the Aedes albopictus mosquito. The extracts of stem and flower of Heliconia rostrata, leaf and stem of Plinia cauliflora, seed of Anonna crassiflora and stem, flower and root of B. pilosa did not present toxicity in the analyzed concentrations. The highest rates of selectivity appeared in the extracts of stem of A. crassiflora and flower of B. pilosa to Staphylococcus aureus, presenting potential for future studies about a new drug development.

Evaluation of attimicrobial and cytotoxic activties of plant extracts from souuthern minas gerais cerrado / Avaliacao das atividades antimicrobiana e citotoxica de extratos de plantas do Cerrado do Sul de Minas Gerais

The antimicrobial activity of plant hidroethanolic extracts on bacteria Gram positive, Gram negative, yeasts, Mycobacterium tuberculosis H37 and Mycobacterium bovis was evaluated by using the technique of Agar diffusion and microdilution in broth. Among the extracts evaluated by Agar diffusion, the extract of Bidens pilosa leaf presented the most expressive average of haloes of growth inhibition to the microorganisms, followed by the extract of B. pilosa flower, of Eugenia pyriformis' leaf and seed, of Plinia cauliflora leaf which statistically presented the same average of haloes inhibitory formation on bacteria Gram positive, Gram negative and yeasts. The extracts of Heliconia rostrata did not present activity. Mycobacterium tuberculosis H37 and Mycobacterium bovis (BCG) appeared resistant to all the extracts. The susceptibility profile of Candida albicans and Saccharomyces cerevisiae fungi were compared to one another and to the Gram positive Bacillus subtilis, Enterococcus faecalis and the Gram negative Salmonella typhimurium bacteria (p > 0.05). The evaluation of cytotoxicity was carried out on C6-36 larvae cells of the Aedes albopictus mosquito. The extracts of stem and flower of Heliconia rostrata, leaf and stem of Plinia cauliflora, seed of Anonna crassiflora and stem, flower and root of B. pilosa did not present toxicity in the analyzed concentrations. The highest rates of selectivity appeared in the extracts of stem of A. crassiflora and flower of B. pilosa to Staphylococcus aureus, presenting potential for future studies about a new drug development.

Foi avaliada a atividade antimicrobiana de extratos hidroetanólicos de plantas sobre bactérias Gram positiva, Gram negativa, leveduras, Mycobacterium tuberculosis H37 e Mycobacterium bovis pela técnica de difusão em Agar e microdiluição em caldo. Dentre os extratos avaliados pelo método de difusão em Agar, o extrato da folha de Bidens pilosa apresentou a mais expressiva média de halos de inibição de crescimento frente aos microrganismos, seguido pelo extrato da flor de B. pilosa, da folha e semente de Eugenia pyriformis, da folha de Plinia cauliflora que apresentaram estatisticamente a mesma média de formação de halos inibitórios sobre bactérias Gram positivas, Gram negativas e leveduras. Os extratos de Heliconia rostrata não apresentaram atividade. Mycobacterium tuberculosis H37 e Mycobacterium bovis (BCG) mostraram-se resistentes a todos os extratos. O perfil de sensibilidade dos fungos Candida albicans e Saccharomyces cerevisiae foram comparáveis entre si e entre as bactérias Gram positivas Bacillus subtilis, Enterococcus faecalis e Gram negativa Salmonella typhimurium (p > 0.05). A avaliação da citotoxicidade foi realizada sobre células C6-36 de larvas de mosquito Aedes albopictus. Os extratos de caule e flor de H. rostrata, folha e caule de P. cauliflora, semente de Anonna crassiflora e caule, flor e raiz de B. pilosa não apresentaram toxicidade nas concentrações avaliadas. Os maiores índices de seletividade foram apresentados pelos extratos de caule de A. crassiflora e flor de B. pilosa para Staphylococcus aureus, apresentando potencial para estudos como futuros candidatos a fármacos.

Avaliação da qualidade higiênico-sanitária dos sucos de laranja comercializados em Alfenas-MG / Evaluation of the hygienic quality of orange juice commercialized in Alfenas-MG

O suco de laranja é um alimento rico em nutrientes e muito consumido em diferentes partes do mundo. A pesquisa de bactérias indicadoras de contaminação auxilia na verificação da qualidade higiênico-sanitária do alimento. As causas mais freqüentes de contaminação dos sucos de laranja são: o controle inadequado da temperatura de estocagem; higiene insatisfatória do ambiente e de maquinários; contaminação cruzada e despreparo do manipulador. Desta forma, o monitoramento microbiológico do produto torna-se muito importante para verificar sua inocuidade. O presente trabalho avaliou a qualidade higiênico-sanitária de sucos de laranja comercializados de Alfenas-MG, comparando os sucos mantidos em refresqueiras com os sucos preparados na hora.(...) De acordo com a RDC n.12/2001, observou-se que 52% das amostras apresentaram NMP de coliformes a 35ºC acima do permitido e 14% das amostras apresentaram NMP de coliformes a 45ºC acima do limite estabelecido pela referida legislação. Observou-se também que os sucos naturais de refresqueiras apresentaram uma maior quantidade de coliformes 35ºC e a 45ºC quando comparados com os sucos naturais preparados na hora.

In vitro evaluation of the antimicrobial activity of endodontic sealers

The purpose of this study is to evaluate the antimicrobial activity of the endodontic sealers: N-Rickert, Sealapex, AH Plus, Mineral Trioxide Aggregate (MTA) and portland cement. The Agar diffusion method was used in plates previously inoculated with the following microorganisms: C. albicans, S. aureus, E. faecalis, E. coli. The diameters of microbial inhibition zones were measured after 24 hours of incubation in kiln at 37°C. According to the methodology used, it was possible to conclude that only the sealers AH Plus and N-Rickert presented antimicrobial activity against C. albicans, S. aureus, and E. coli; no antimicrobial activity in MTA, Sealapex and portland cement was observed. N-Rickert presented the largest inhibition zones varying from 8 to 18 mm, and the microorganism E. faecalis was resistant against all sealers tested.

O objetivo deste estudo é avaliar a capacidade antimicrobiana dos cimentos obturadores de canal: N-Rickert, Sealapex, AH Plus e também do Agregado Trióxido Mineral (MTA) e Cimento de Portland. O método utilizado foi a difusão em Agar, em placas previamente inoculadas com os seguintes microorganismos: C. albicans, S. aureus, E. faecalis, E. coli. A leitura do diâmetro do halo de inibição do crescimento microbiano foi realizada, após 24 horas de incubação, em estufa a 37°C. De acordo com a metodologia empregada, foi possível concluir que: somente os cimentos obturadores AH Plus e N-Rickert apresentaram atividade antimicrobiana contra C. albicans, S. aureus e E. coli; não foi observada atividade antimicrobiana no cimento de Portland, MTA e Sealapex. O cimento N-Rickert apresentou halos de inibição maiores variando de 8 a 18 mm. O microorganismo E. faecalis foi resistente contra todos os cimentos testados.

Molecular identification of Candida dubliniensis isolated from oral lesions of HIV-positive and HIV-negative patients in São Paulo, Brazil.

Candida dubliniensis is a new, recently described species of yeast. This emerging oral pathogen shares many phenotypic and biochemical characteristics with C. albicans, making it hard to differentiate between them, although they are genotypically distinct. In this study, PCR (Polymerase Chain Reaction) was used to investigate the presence of C. dubliniensis in samples in a culture collection, which had been isolated from HIV-positive and HIV-negative patients with oral erythematous candidiasis. From a total of 37 samples previously identified as C. albicans by the classical method, two samples of C. dubliniensis (5.4%) were found through the use of PCR. This study underscores the presence of C. dubliniensis, whose geographical and epidemiological distribution should be more fully investigated.

Molecular identification of Candida dubliniensis isolated from oral lesions of HIV-positive and HIV-negative patients in São Paulo, Brazil

Candida dubliniensis é uma nova espécie recentemente descrita. Este patógeno oral emergente compartilha muitas características fenotípicas e bioquímicas com C. albicans dificultando assim a diferenciação entre elas. As mesmas, porém, mostram-se genotipicamente distintas. Este trabalho tem como objetivo identificar, pela técnica de PCR (Polymerase Chain Reaction), a possível presença de C. dubliniensis dentre amostras isoladas de candidose oral eritematosa, provenientes de pacientes HIV positivos e HIV negativos. Em um total de 37 amostras identificadas anteriormente, por método clássico, como C. albicans encontramos duas amostras de C. dubliniensis (5,4%) utilizando a técnica do PCR. Esta técnica mostrou-se útil, prática e com identificação taxonômica mais acurada.

In vitro evaluation of the antimicrobial activity of endodontic sealers.

The purpose of this study is to evaluate the antimicrobial activity of the endodontic sealers: N-Rickert, Sealapex, AH Plus, Mineral Trioxide Aggregate (MTA) and portland cement. The Agar diffusion method was used in plates previously inoculated with the following microorganisms: C. albicans, S. aureus, E. faecalis, E. coli. The diameters of microbial inhibition zones were measured after 24 hours of incubation in kiln at 37 degrees C. According to the methodology used, it was possible to conclude that only the sealers AH Plus and N-Rickert presented antimicrobial activity against C. albicans, S. aureus, and E. coli; no antimicrobial activity in MTA, Sealapex and portland cement was observed. N-Rickert presented the largest inhibition zones varying from 8 to 18 mm, and the microorganism E. faecalis was resistant against all sealers tested.