The role of IL-10 in immune responses against Pseudomonas aeruginosa during acute lung infection.

Pseudomonas aeruginosa is considered an opportunistic pathogen of great clinical importance. The clearance of this bacterium occurs through recognition of the pathogen by innate immune system receptors, leading to a lung inflammatory response. However, this response must be controlled via immunoregulatory pathways. In this study, we evaluate the role of endogenous murine IL-10 after acute infection with the virulent strain P. aeruginosa PA14. To assess the role of IL-10, intratracheal infection with the PA14 strain was performed in C57BL/6 or IL-10 KO mice. The PA14 strain was recovered in both types of animals, although IL-10 KO mice presented a higher number of viable bacteria in the lung when compared to the C57BL/6 group. Histopathological and stereological analyses showed that IL-10 KO mice had higher tissue damage and inflammatory infiltrate when compared to control animals. The activity of MMP-9 but not MMP-2, as well as IL-6 and TNF-α expression, were augmented in the lungs of infected animals and was much more evident in IL-10 KO animals when compared to the other analyzed groups. This work indicates that endogenous IL-10 control P. aeruginosa infection, the expression of pro-inflammatory genes, MMP-9 activity and histopathological processes of the infectious process in question.

Anti-inflammatory and antinociceptive activities of a phenylpropanoid-enriched fraction of Duguetia furfuracea.

A previous study reported the in vivo anti-inflammatory and antinociceptive activities of essential oil of the underground stem bark of Duguetia furfuracea, termed EODf. This study aimed to obtain a phenylpropanoid-enriched fraction from the D. furfuracea (EFDf) essential oil and to investigate its anti-inflammatory and antinociceptive effects. The chemical composition of the EFDf was determined by gas chromatography-mass spectrometry (GC-MS). The in vivo anti-inflammatory activity was evaluated with a lipopolysaccharide (LPS)-induced paw oedema model. The effects of the EFDf on the polymorphonuclear leukocyte recruitment and the inducible nitric oxide synthase (iNOS) expression were evaluated in mice footpads. Moreover, the in vivo antinociceptive effect was assayed using the formalin test and the LPS-induced thermal hyperalgesia model. In the EFDf, 8 major compounds were identified, with α-asarone (36.4%) and 2,4,5-trimethoxystyrene (27.8%) the main constituents. A higher concentration of phenylpropanoid derivatives was found in the EFDf, 64.2% compared to the EODf (38%). The oral (p.o.) treatment with the EFDf at a dose of 3 mg/kg significantly attenuated the paw oedema, polymorphonuclear leukocyte migration, iNOS expression, and tumour necrosis factor alpha (TNF-α) production. The EFDf (10 and 30 mg/kg) also inhibited both phases of the formalin test and caused a significant increase in the reaction time in the LPS-induced thermal hyperalgesia model. Finally, EFDf-treated animals did not show any alteration of motor coordination. The results suggest that the enrichment of 2,4,5-trimethoxystyrene and α-asarone enhances the anti-inflammatory activity of the EFDf compared to the EODf. In contrast, the antinociception promoted by the EFDf was similar to the EODf and was mediated via activation of adenosinergic and opioidergic receptors.

Anti-inflammatory and central and peripheral anti-nociceptive activities of α-asarone through the inhibition of TNF-α production, leukocyte recruitment and iNOS expression, and participation of the adenosinergic and opioidergic systems.

Alpha-asarone has been found to possess many pharmacological activities, which can improve cognitive function and exert anti-oxidant, anxiolytic, anti-epileptic and protective effects against endothelial cell injury. The anti-inflammatory activity of α-asarone was evaluated using lipopolysaccharide (LPS)-induced paw oedema. Moreover, leukocyte migration, inducible nitric oxide synthase (iNOS) expression and tumour necrosis factor-alpha (TNF-α) levels were quantified in footpads. Formalin and LPS-induced thermal hyperalgesia models were generated using adenosinergic, opioidergic, serotonergic and muscarinic receptor antagonists. The effects on motor coordination were evaluated by means of the rota-rod test. Oral treatment (p.o.) with α-asarone (3 mg/kg) significantly inhibited paw oedema by 62.12 and 72.22%, 2 and 4 h post LPS injection, respectively. Alpha-asarone (3 mg/kg, p.o.) attenuated the inflammatory infiltrate 1, 3 and 6 h after LPS injection. Furthermore, α-asarone (3 mg/kg, p.o.) suppressed iNOS expression and TNF-α production, 6 and 1 h after inflammatory stimulus, respectively. Alpha-asarone (3, 10 and 30 mg/kg, p.o.) inhibited both phases of formalin-induced licking. In the hot-plate test, α-asarone (10 and 30 mg/kg, p.o.) increased the latency to response 3 and 5 h post LPS stimulus. Caffeine and naloxone abolished the central anti-nociceptive effect of α-asarone (neurogenic phase of formalin and hot plate tests), suggesting the participation of the adenosinergic and opioidergic systems. Furthermore, naloxone reversed the peripheral activity of α-asarone (inflammatory phase of formalin test), indicating the possible involvement of the opioidergic pathway. In the rota-rod test, α-asarone did not change motor coordination. These findings suggest that α-asarone has anti-inflammatory, peripheral and central anti-nociceptive effects and could represent a promising agent for future research.

Chemical composition and evaluation of the anti-inflammatory and antinociceptive activities of Duguetia furfuracea essential oil: Effect on edema, leukocyte recruitment, tumor necrosis factor alpha production, iNOS expression, and adenosinergic and opioidergic systems.

ETHNOPHARMACOLOGICAL RELEVANCE: Duguetia furfuracea (A. St. -Hil.) Saff. (Annonaceae) is commonly known in Brazil as "araticum-seco," and its root is used in folk medicine to treat inflammatory and painful disorders. However, no studies have been performed to evaluate these therapeutic activities. AIM OF THE STUDY: Investigate the chemical composition, anti-inflammatory and antinociceptive effects, and elucidate the possible antinociceptive mechanisms of action from the essential oil of D. furfuracea (EODf) underground stem bark. MATERIALS AND METHODS: Chemical composition was determined by gas chromatography and mass spectrometry (GC/MS). The paw edema induced by LPS, formalin-induced nociception, LPS-induced thermal hyperalgesia and rota-rod tests in vivo were used to evaluate the anti-inflammatory and antinociceptive effects in addition to the alteration on motor coordination. Histological analyses and an immunohistochemistry assay for iNOS were performed on mouse footpads of naive, control, 10 mg/kg EODf, and 10 mg/kg indomethacin (Ind) groups. The samples were removed at 1, 3, and 6 h after subplantar injection of LPS. In addition, the involvement of the adenosinergic, opioidergic, serotonergic, and cholinergic systems were investigated, in order to elucidate possible antinociceptive mechanisms. RESULTS: Twenty-four volatile constituents were detected and identified. (E)-asarone (21.9%), bicyclogermacrene (16.7%), 2,4,5-trimethoxystyrene (16.1%), α-gurjunene (15%), cyperene (7.8%), and (E)-caryophyllene (4.6%) were major compounds found in EODf. Oral treatment (p.o.) with EODf (1, 3, and 10 mg/kg) significantly inhibited the paw edema induced by LPS. At 10 mg/kg EODf promoted inhibition of tumor necrosis factor alpha (TNF-α) production, recruitment of polymorphonuclear (PMN) leukocytes and inducible nitric oxide synthase (iNOS) expression in paw tissue. EODf (10 and 30 mg/kg, p.o.) also reduced licking time in both phases of the formalin test and it had a significant effect on the LPS-induced thermal hyperalgesia model. The administration of caffeine (Caf) and naloxone (Nal) reversed the antinociceptive activity of EODf, in the first phase of the formalin test and in the LPS-induced thermal hyperalgesia model. Moreover, Nal was also able to abolish the antinociception caused by EODf, in the second phase of formalin test. In the rota-rod test, EODf-treated animals did not show any alteration of motor coordination. CONCLUSIONS: Our findings indicate that EODf underground stem bark produces anti-inflammatory and both central and peripheral antinociceptive effects. Furthermore, the antinociceptive activity of EODf underground stem bark is possibly mediated by adenosinergic and opioidergic pathways, and its properties do not induce effects on motor coordination. These results support the use of the folk medicine, D. furfuracea root, to treat inflammation and painful conditions.

Clinical, Molecular, and Epidemiological Analysis of Dengue Cases during a Major Outbreak in the Midwest Region of Minas Gerais, Brazil.

This study aims to perform the first molecular and clinical-epidemiological analysis of dengue cases in Divinopolis, MG, Brazil. Data from 4,110 cases of dengue were accessed and 190 clinical samples were collected for molecular analyses. In this study, 2.7% of the men and 3.0% of the women were admitted to hospital. There was no association between gender and hospital admission. The symptoms observed in this study are according to the Health Ministry, but fever was present in 82.2% and not in 100% of cases. The chance of hospital admission was 1.55 higher in patients with any kind of bleeding (334) and 2.4% of individuals without bleeding were also hospitalized due to other warning signs. In the molecular analyses, 23% of the samples were positive for DENV. DENV-2 and DENV-3 were identified in 2010, DENV-3 in 2011, DENV-1 in 2012, and DENV-1 and DENV-4 in 2013. DENV detection was possible in samples with only one day of symptoms. This first report of dengue data in Divinópolis provided more insight into the viral types and effects of disease in the city, confirming the need for caution in assessing cases of suspected dengue and for revision of the criteria proposed by the Health Ministry to classify cases of the disease.

Ocorrência de esquistossomose em Divinópolis-MG baseada em estudo com escolares e em inquéritos de notificação da doença / Occurrence of schistosomiasis in Divinópolis-MG based on study of schoolchildren and surveys of disease notification

Introduction: Schistosomiasis is a neglected chronic disease that affects mainly underdeveloped regions, including Brazil. Objective: To evaluate the distribution profile of the schistosomiasis in Divinópolis-MG. Material and methods: It is characterized as a descriptive and analytical epidemiological study. A parasitological study performed in schoolchildren of public municipal schools; simultaneously, a survey of schistosomiasis cases reported in the city between 2005 and 2011 years was performed with the Municipal Department of Health. Data related to the characteristics of the infection were observed, such as: affected areas, age, gender, and professional occupation. Results: This survey showed 33 cases of schistosomiasis in the city during this period, which the most of them (84.8%) were between 20-59 years of age. The results of the study with the schoolchildren are in agreement with those obtained through the reporting forms, both indicating no occurrence of schistosomiasis in individuals between 6-14 years of age in Divinópolis. Conclusion: The absence of the disease in children and adolescents analyzed and the presence in adults is a strong evidence of exogenous contamination in the city, especially as a result of immigration or rural tourism, and possible changes in habits, related to risk factors...

Introdução: A esquistossomose é uma doença crônica negligenciada, que afeta principalmente regiões subdesenvolvidas, incluindo o Brasil. Objetivo: Avaliar o perfil de distribuição da esquistossomose na região de Divinópolis-MG. Material e métodos: Caracteriza-se como um estudo epidemiológico analítico e descritivo. Foi realizado um estudo parasitológico em escolares da rede pública municipal e, concomitantemente, realizou-se também um levantamento dos casos notificados da doença no município entre 2005 e 2011, junto à Secretaria Municipal de Saúde. Foram verificados dados relacionados com as características da infecção, como regiões afetadas, faixa etária, gênero e ocupação profissional. Resultados: O levantamento apontou 33 casos de esquistossomose no município neste período, sendo a maior parte deles (84,8%) na faixa etária de 20 a 59 anos. Os resultados do estudo com os escolares estão em concordância com os obtidos por meio das fichas de notificação, ambos indicando a não ocorrência de esquistossomose em indivíduos de 6 a 14 anos em Divinópolis. Conclusão: A ausência da doença nas crianças e nos adolescentes analisados e a presença em adultos sugerem forte evidência de contaminação exógena no município, especialmente fruto de imigração ou turismo rural, além de possíveis mudanças de hábitos, relacionados com os fatores de risco...

Cervical uterine lesions: an epidemiological and molecular investigation in midwestern Minas Gerais, Brazil.

Cervical uterine cancer is the second most frequent form of cancer in the female population and the fourth cause of cancer deaths among Brazilian women. In Divinópolis county, Minas Gerais state, cervical cancer accounted for 6.6% of deaths in 2007. The purpose of this study was to conduct an epidemiological and molecular investigation of samples of pre-neoplastic and neoplastic cervical uterine lesions from patients assisted at public healthcare services in the county. The study was based on clinical-epidemiological descriptions retrieved from cytological examination request forms from 2006 to 2010. For molecular analysis, samples from 95 patients were collected and DNA was extracted using the Chelex 100 method. PCR was performed for detection and typing of HPV 6/11, 16, 18, 31, and 33. Of the 52 168 patients who underwent cytological examination, 625 had pre-neoplastic or neoplastic lesions during the study period. Age distribution was consistent with the literature, with a predominance of patients aged 20-49 years. The microorganism found most frequently was Lactobacillus sp. (65%). Prominent among cellular alterations were cervical intraepithelial neoplasia I (39.7%) and atypical squamous cells of undetermined significance (32%). Molecular analysis revealed 72.6% of positive samples for HPV. HPV 16 (26.3%) was the most frequent type, followed by types 6/11 (7.4%), 18 (5.3%), 33 (2.1%), and 31 (1%). The results provided improved understanding of the association between HPV and cancer in Divinópolis, in addition to providing data that can contribute to the design of measures to prevent and control HPV infection in the county investigated.

Biochemical studies with DNA polymerase beta and DNA polymerase beta-PAK of Trypanosoma cruzi suggest the involvement of these proteins in mitochondrial DNA maintenance.

Mammalian DNA polymerase beta is a nuclear enzyme involved in the base excision and single-stranded DNA break repair pathways. In trypanosomatids, this protein does not have a defined cellular localization, and its function is poorly understood. We characterized two Trypanosoma cruzi proteins homologous to mammalian DNA polymerasebeta, TcPolbeta and TcPolbetaPAK, and showed that both enzymes localize to the parasite kinetoplast. In vitro assays with purified proteins showed that they have DNA polymerization and deoxyribose phosphate lyase activities. Optimal conditions for polymerization were different for each protein with respect to dNTP concentration and temperature, and TcPolbetaPAK, in comparison to TcPolbeta, conducted DNA synthesis over a much broader pH range. TcPolbeta was unable to carry out mismatch extension or DNA synthesis across 8-oxodG lesions, and was able to discriminate between dNTP and ddNTP. These specific abilities of TcPolbeta were not observed for TcPolbetaPAK or other X family members, and are not due to a phenylalanine residue at position 395 in the C-terminal region of TcPolbeta, as assessed by a site-directed mutagenesis experiment reversing this residue to a well conserved tyrosine. Our data suggest that both polymerases from T. cruzi could cooperate to maintain mitochondrial DNA integrity through their multiple roles in base excision repair, gap filling and translesion synthesis.

Escherichia coli as a model system to study DNA repair genes of eukaryotic organisms.

The bacteria Escherichia coli has been widely employed in studies of eukaryotic DNA repair genes. Several eukaryotic genes have been cloned by functional complementation of mutant lineages of E. coli. We examined the similarities and differences among bacterial and eukaryotic DNA repair systems. Based on these data, we examined tools used for gene cloning and functional studies of DNA repair in eukaryotes, using this bacterial system as a model.