Quorum sensing detected by atomic force microscopy imaging of corrals surrounding multicellular arrangement of bacteria

Connectivity of the glycocalyx covering of small communities of Acidithiobacillus ferrooxidans bacteria deposited on hydrophilic mica plates was imaged by atomic force microscopy. When part of the coverage was removed by water rinsing, an insoluble structure formed by corrals surrounding each individual bacterium was observed. A collective ring structure with clustered bacteria (>= 3) was observed, which indicates that the bacteria perceived the neighborhood in order to grow a protective structure that results in smaller production of exopolysaccharides material. The most surprising aspect of these collective corral structures was that they occur at a low bacterial cell density. The deposited layers were also analyzed by confocal Raman microscopy and shown to contain polysaccharides, protein, and glucoronic acid.

Novel insights into the genomic basis of citrus canker based on the genome sequences of two strains of Xanthomonas fuscans subsp aurantifolii

Background: Citrus canker is a disease that has severe economic impact on the citrus industry worldwide. There are three types of canker, called A, B, and C. The three types have different phenotypes and affect different citrus species. The causative agent for type A is Xanthomonas citri subsp. citri, whose genome sequence was made available in 2002. Xanthomonas fuscans subsp. aurantifolii strain B causes canker B and Xanthomonas fuscans subsp. aurantifolii strain C causes canker C. Results: We have sequenced the genomes of strains B and C to draft status. We have compared their genomic content to X. citri subsp. citri and to other Xanthomonas genomes, with special emphasis on type III secreted effector repertoires. In addition to pthA, already known to be present in all three citrus canker strains, two additional effector genes, xopE3 and xopAI, are also present in all three strains and are both located on the same putative genomic island. These two effector genes, along with one other effector-like gene in the same region, are thus good candidates for being pathogenicity factors on citrus. Numerous gene content differences also exist between the three cankers strains, which can be correlated with their different virulence and host range. Particular attention was placed on the analysis of genes involved in biofilm formation and quorum sensing, type IV secretion, flagellum synthesis and motility, lipopolysacharide synthesis, and on the gene xacPNP, which codes for a natriuretic protein. Conclusion: We have uncovered numerous commonalities and differences in gene content between the genomes of the pathogenic agents causing citrus canker A, B, and C and other Xanthomonas genomes. Molecular genetics can now be employed to determine the role of these genes in plant-microbe interactions. The gained knowledge will be instrumental for improving citrus canker control.

Long-Chain Acyl-Homoserine Lactones from Methylobacterium mesophilicum: Synthesis and Absolute Configuration

The acyl-homoserine lactones (acyl-HSLs) produced by Methylobacterium mesophilicum isolated from orange trees infected with the citrus variegated chlorosis (CVC) disease have been studied, revealing the occurrence of six long-chain acyl-HSLs, i.e., the saturated homologues (S)-N-dodecanoyl (1) and (S)-N-tetradecanoyl-HSL (5), the uncommon odd-chain N-tridecanoyl-HSL (3), the new natural product (S)-N-(2E)-dodecenoyl-HSL (2), and the rare unsaturated homologues (S)-N-(7Z)-tetradecenoyl (4) and (S)-N-(2E,7Z)-tetradecadienyl-HSL (6). The absolute configurations of all HSLs were determined as 3S. Compounds 2 and 6 were synthesized for the first time. Antimicrobial assays with synthetic acyl-HSLs against Gram-positive bacterial endophytes co-isolated with M. mesophilicum from CVC-infected trees revealed low or no antibacterial activity.

a-SiC : H anti-resonant layer ARROW waveguides

Over the last decades, anti-resonant reflecting optical waveguides (ARROW) have been used in different integrated optics applications. In this type of waveguide, light confinement is partially achieved through an anti-resonant reflection. In this work, the simulation, fabrication and characterization of ARROW waveguides using dielectric films deposited by a plasma-enhanced chemical vapor deposition (PECVD) technique, at low temperatures(similar to 300 degrees C), are presented. Silicon oxynitride (SiO(x)N(y)) films were used as core and second cladding layers and amorphous hydrogenated silicon carbide(a-SiC:H) films as first cladding layer. Furthermore, numerical simulations were performed using homemade routines based on two computational methods: the transfer matrix method (TMM) for the determination of the optimum thickness of the Fabry-Perot layers; and the non-uniform finite difference method (NU-FDM) for 2D design and determination of the maximum width that yields single-mode operation. The utilization of a silicon carbide anti-resonant layer resulted in low optical attenuations, which is due to the high refractive index difference between the core and this layer. Finally, for comparison purposes, optical waveguides using titanium oxide (TiO(2)) as the first ARROW layer were also fabricated and characterized.

Anti-oxidative and anti-ulcerogenic activity of Ipomoea imperati

Ipomoea imperati (Vahl) Griseb., Convolvulaceae, is used in traditional medicine for the treatment of inflammation, swelling and wounds, as well as to treat pains and stomach problems. This work evaluates the anti-oxidative activity by ESR (Electron Spin Resonance spectroscopy) and the preventive and curative actions of I. imperati in gastric ulcer animal model. Ipomoea imperati (200 mg/kg, p.o.) prevented the formation of gastric lesions in 78% (p<0.05) when compared with the negative control tween 80. Lanzoprazole, prevented in 85% the gastric lesions formation induced by ethanol (p<0.05). Therefore, the oral administration of I. imperati one hour before the ulcerogenic agent prevented the ulcer formation, conserving the citoprotection characteristics of the gastric mucosa and assuring the integrity of gastric glands and gastric fossets. The healing activity of I. imperati (200 mg/kg, p.o.) evaluated in chronic ulcer experiments induced by the acetic acid, was 72% (p<0.05). The positive control, ranitidine, healed 78% of the gastric lesions (p<0.05). The histological analysis confirmed the recovery of the mucosal layer and the muscle mucosal layer harmed by the acetic acid. Experiments in vitro with DPPH (2.2-diphenyl-1-picrylhydrazyl) of anti-oxidative activity demonstrated that I. imperati presents an IC50 of 0.73±0.01 mg/mL.

Caracterização físico-química de polpas de frutos da Amazônia e sua correlação com a atividade anti-radical livre

Características físico-químicas (cor, pH, acidez total titulável, sólidos solúveis totais, conteúdo de lipídios e umidade) e níveis de compostos bioativos (ácido ascórbico, fenólicos totais) foram determinados em quinze amostras de polpas de frutos procedentes da região Amazônica (abiu, acerola, açaí, araçá-boi, bacaba, bacuri, buriti, cajá, cajarana, caju, cupuaçu, graviola, murici, noni e tamarindo). A atividade de radicais livres foi avaliada pelo método de ABTS. Algumas polpas apresentaram alta potencialidade antioxidante, associada com a atividade antirradicais livres obtida e os conteúdos dos componentes bioativos como compostos fenólicos e ácido ascórbico, destacando-se acerola e acaí. O conteúdo total de compostos fenólicos foi correlacionado à capacidade antioxidante das polpas.

Validação e reprodutibilidade de uma escala de auto-eficácia para adesão ao tratamento anti-retroviral em pais ou cuidadores de crianças e adolescentes vivendo com HIV/AIDS

OBJETIVO: Validar uma escala de auto-eficácia para adesão ao tratamento anti-retroviral em crianças e adolescentes com HIV/AIDS, levando em consideração a perspectiva dos pais/responsáveis, e avaliar a sua reprodutibilidade. MÉTODOS: O estudo foi realizado no Hospital-Dia do Centro de Referência e Treinamento em DST/AIDS de São Paulo. Foram entrevistados os pais/responsáveis de 54 crianças e adolescentes de 6 meses a 20 anos que passaram em consulta de rotina pelo serviço. Os dados de auto-eficácia foram levantados pela escala de auto-eficácia para seguir prescrição anti-retroviral (AE), que foi calculada de duas maneiras: análise fatorial e fórmula já definida. A consistência interna da escala foi verificada pelo coeficiente ade Cronbach. A validade foi avaliada pela comparação das médias dos escores entre grupos de pacientes aderentes e não aderentes ao tratamento anti-retroviral (teste de Mann-Whitney) e cálculo do coeficiente de correlação de Spearman entre os escores e parâmetros clínicos. A reprodutibilidade foi verificada por meio do teste de Wilcoxon, pelo coeficiente de correlação intraclasse (CCI) e pelo gráfico de Bland-Altman. RESULTADOS: A escala de AE apresentou boa consistência interna (a= 0,87) e boa reprodutibilidade (CCI = 0,69 e CCI = 0,75). Quanto à validade, a escala de AE conseguiu discriminar pacientes aderentes e não aderentes ao tratamento anti-retroviral (p = 0,002) e apresentou correlação significativa com a contagem de CD4 (r = 0,28; p = 0,04). CONCLUSÕES: A escala de AE pode ser utilizada para avaliar a adesão à terapia anti-retroviral em crianças e adolescentes com HIV/AIDS, levando em consideração a perspectiva dos pais/cuidadores.

Auto-anticorpos anti-LDLox e sua correlação com o perfil lipídico e o estado nutricional de adolescentes

OBJETIVO: Avaliar se o conteúdo de auto-anticorpos anti-LDL oxidada (anti-LDLox) no plasma de adolescentes correlaciona-se com suas medidas antropométricas e com o perfil lipídico. MÉTODOS: O estudo incluiu 150 adolescentes com idade entre 10 e 15 anos, recrutados do ambulatório de obesidade da Universidade Federal de São Paulo (SP) e de escolas públicas de Piracicaba (SP). Foram avaliadas medidas antropométricas, como índice de massa corporal, circunferência de cintura e do braço, classificando os adolescentes em eutrófico, sobrepeso e obeso. Para as análises bioquímicas, foi realizado o perfil lipídico através de métodos enzimáticos colorimétricos, e para detecção do conteúdo de auto-anticorpos anti-LDLox, utilizou-se o método de ELISA. RESULTADOS: Segundo análises das variáveis antropométricas, o grupo obeso apresentou perfil alterado em relação aos grupos eutrófico e sobrepeso (p < 0,01), indicando risco cardiovascular. Quando o perfil lipídico foi avaliado, observaram-se diferenças estatisticamente significativas para as concentrações de colesterol total (p = 0,011), HDL-colesterol (p = 0,001) e LDL-colesterol (p < 0,042) nos grupos eutrófico e obeso. Para as análises de auto-anticorpos anti-LDLox plasmática, os grupos sobrepeso (p = 0,012) e obeso (p < 0,001) apresentaram valores superiores ao grupo eutrófico. Também houve correlações entre os auto-anticorpos anti-LDLox e variáveis antropométricas. CONCLUSÃO: A presença de auto-anticorpos anti-LDLox em adolescentes e as alterações metabólicas no perfil lipídico variaram de modo proporcional com parâmetros antropométricos, o que torna o conteúdo de anti-LDLox um potencial indicador bioquímico de risco para síndrome metabólica.

Detection of anti-Toxoplasma gondii antibodies in experimentally and naturally infected non-human primates by Indirect Fluorescence Assay (IFA) and indirect ELISA

The Indirect Fluorescence Assay (IFA) and the indirect ELISA were comparatively used to detect IgG and IgM antibodies for Toxoplasma gondii in experimentally and naturally infected primates. In the experimentally infected group, antibodies of diagnostic value were detected at day 9 post-infection (PI) with the IFA (IgG and IgM) and with IgG-ELISA. IgM-ELISA detected antibodies for T. gondii starting at day 3 PI until the end of the experiment (102 days PI). Of the 209 naturally infected sera tested, from many zoos of State of Sao Paulo, 64.59 and 67.94% were positive in the IgG-IFA test and IgG-ELISA respectively. IgM-ELISA test detected seropositivity in 52.63% of the sera although IgM-IFA test detected it in only in 0.96% of the samples. The differential toxoplasmosis diagnosis was accomplished with Neospora caninum by IFA, observing 61 (29.2%) seropositive animals for this parasite and 149 (70.8%) negative. Sixty animals were positive for both T. gondii and N. caninum. Pneumonia, splenomegaly, and intestinal ulcers were macroscopically observed. Unremarkable interstitial pneumonia, enteritis, colitis, splenitis, and glomerulitis were microscopically observed. The immunohistochemical stain could not detect the presence of T. gondii in the tissues of the animals infected experimentally.

Seroprevalence of anti-Leishmania spp. antibodies in rural dogs from the city of Monte Negro, State of Rondônia, Brazil

The present study assessed the prevalence of anti-Leishmania spp. antibodies in dogs from the city of Monte Negro, State of Rondônia, Brazil. ELISA (NE > 3) and IFAT (>1:40) were used to evaluate 161 serum samples collected from rural dogs from Monte Negro. Forty-five (27.9%) dogs were positive by ELISA tests and five (3.1%) were positive by IFAT. The present study showed for the first time the frequency of exposure to Leishmania spp. in dogs in the State of Rondônia, Amazon Region.

Prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in swine from Northeastern Brazil

A serologic survey was conducted among 130 swine slaughtered in the public slaughterhouse of the city of Patos, Paraíba State, Northeastern Brazil, to determine the prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies, and to verify possible associations between sex of the animals and antibody prevalence. The sera were analyzed by indirect antibody tests, considering 1:64 (T. gondii) and 1:50 (N. caninum) dilutions as cut-off points. The prevalence of anti-T. gondii antibodies was 36.2% (47/130) (95% CI = 27.9 - 45.0%) with reciprocal titers ranging from 64 to 2,048, and of anti-N. caninum antibodies was 3.1% (4/130) (95% CI = 0.8 - 7.7%) with reciprocal titers ranging from 50 to 6,400. Three of the four N. caninum-positive samples were also positive for T. gondii antibodies. All Neospora and Toxoplasma IFAT-positive animals were also positive for confirmatory immunoblotting techniques using total and purified N. caninum and T. gondii tachyzoite antigens, i.e., p38 (NcSRS2) and p30 (TgSAG1). There was no association between sex of animals and prevalence of anti-T. gondii and anti-N. caninum antibodies. This is the first indication of N. caninum natural infection in pigs from Brazil.

Occurrence of anti-Neospora caninum and anti-Toxoplasma gondii antibodies in dogs with visceral leishmaniasis

Uninfected dogs and those naturally infected with Leishmania chagasi exhibiting different clinical forms of disease were evaluated for the presence of anti-Neospora caninum and anti-Toxoplasma gondii antibodies. Blood samples were collected from 110 mongrel dogs. Sera were tested using the indirect fluorescent antibody test (IFAT), and the animals with visceral leishmaniasis (VL) (n=60) were classified clinically. Out of the 110 sera investigated, 5 (4.5%) were positive for N. caninum (IFAT≥50) and 36 (32.7%) for T. gondii (IFAT≥16). Anti-L. chagasi antibody titers in asymptomatic dogs (n=10) were found to be significantly lower (P<0.05) than those in oligosymptomatic ones (n=22), which were in turn significantly lower (P<0.05) than those in symptomatic ones (n=28). No association between Leishmania and N. caninum infections was observed. Among dogs infected with L. chagasi, a tendency (P=0.053) towards an association between the infection with T. gondii and the appearance of VL symptoms was observed, suggesting that the clinical manifestation of VL in dogs may enhance their susceptibility to T. gondii. The possible influence of the immunosuppressive status of canine leishmaniasis in the different clinical forms of the disease is discussed.

Prevalência de anticorpos anti-Chlamydophila spp. em propriedades rurais com histórico de aborto bovino no estado do Paraná

Chlamydophila abortus é o agente etiológico do aborto epizoótico bovino, cujas manifestações clínicas mais freqüentes são aborto, nascimento de bezerros prematuros e de animais fracos, natimorto e repetição de cio em intervalos irregulares. O objetivo deste trabalho foi estimar a prevalência de anticorpos anti-Chlamydophila spp. em fêmeas bovinas de propriedades rurais com histórico de aborto, selecionadas dentro do delineamento amostral do Plano Nacional de Controle e Erradicação da Brucelose e Tuberculose no estado do Paraná. Foram testadas pela prova de fixação de complemento 3.102 amostras de soro de fêmeas bovinas (idade > 24 meses), provenientes de 373 propriedades. Ao total, 44 (1,42%) animais foram positivos com títulos > 32. A prevalência de focos foi de 8,82% (6,15%-12,17%). Animais confinados ou semi-confinados (OR=3.339, P=0.004), propriedade com menos de 35 matrizes (OR=3.339, P=0.017), presença de produtos do aborto na pastagem (OR=2.372, P=0.037) e aluguel de pasto (OR=3.398, P=0.006) foram considerados fatores de risco para Chlamydophila spp. A infecção por Chlamydophila spp. acometeu um número pequeno de animais, oriundos de propriedades com histórico de aborto. A importância deste agente como causa de aborto em bovinos no estado do Paraná, se existir, é muito pequena.

Avaliação da ocorrência de anticorpos anti-Toxoplasma gondii, em soros de caprinos do estado de São Paulo, e associação com variáveis epidemiológicas, problemas reprodutivos e riscos à saúde pública

Objetivando avaliar a ocorrência de anticorpos anti-Toxoplasma gondii, em caprinos leiteiros do estado de São Paulo, e verificar possíveis associações com idade, sexo, presença de gatos, problemas reprodutivos e potenciais riscos à saúde pública, foram considerados soros de 923 caprinos, de ambos os sexos e idade acima de três meses, provenientes de 17 propriedades de diferentes municípios. Para o diagnóstico, utilizou-se a reação de imunofluorescência indireta (RIFIe"16) e, também, um inquérito sobre saúde, a fim de se coletarem informações epidemiológicas e de esfera reprodutiva de todos os capris. Os resultados foram discutidos no nível de 5% de significância. Do total das 17 propriedades, foram diagnosticados 15 focos de T. gondii, com positividade entre 2,70% e 81,25%. Não foram verificadas associações entre freqüência de soropositividade e sexo dos animais nem ocorrência de falhas reprodutivas, nos capris. Constatou-se influência positiva na taxa de anticorpos anti-T. gondii pelo aumento da idade dos caprinos e presença de gatos, nos capris. Além de a enfermidade encontrar-se amplamente difundida no estado de São Paulo, o risco eminente de transmissão de T. gondii à saúde pública também deve ser considerado, uma vez que se encontraram focos onde se comercializavam produtos "in natura", como leite e carne.