A combined approach for comparative exoproteome analysis of Corynebacterium pseudotuberculosis

Background: Bacterial exported proteins represent key components of the host-pathogen interplay. Hence, we sought to implement a combined approach for characterizing the entire exoproteome of the pathogenic bacterium Corynebacterium pseudotuberculosis, the etiological agent of caseous lymphadenitis (CLA) in sheep and goats. Results: An optimized protocol of three-phase partitioning (TPP) was used to obtain the C. pseudotuberculosis exoproteins, and a newly introduced method of data-independent MS acquisition (LC-MSE) was employed for protein identification and label-free quantification. Additionally, the recently developed tool SurfG+ was used for in silico prediction of sub-cellular localization of the identified proteins. In total, 93 different extracellular proteins of C. pseudotuberculosis were identified with high confidence by this strategy; 44 proteins were commonly identified in two different strains, isolated from distinct hosts, then composing a core C. pseudotuberculosis exoproteome. Analysis with the SurfG+ tool showed that more than 75% (70/93) of the identified proteins could be predicted as containing signals for active exportation. Moreover, evidence could be found for probable non-classical export of most of the remaining proteins. Conclusions: Comparative analyses of the exoproteomes of two C. pseudotuberculosis strains, in addition to comparison with other experimentally determined corynebacterial exoproteomes, were helpful to gain novel insights into the contribution of the exported proteins in the virulence of this bacterium. The results presented here compose the most comprehensive coverage of the exoproteome of a corynebacterial species so far

Experimental periodontitis promotes transient vascular inflammation and endothelial dysfunction

AbstractObjectivesThis study aimed to evaluate the systemic inflammatory response and cardiovascular changes induced by experimental periodontitis in rats.DesignExperimental periodontitis was induced by placing a cotton ligature around the cervix of both sides of mandibular first molars and maxillary second molars in each male rat. Sham-operated rats had the ligature removed immediately after the procedure. Seven, 14 or 28 days after procedure, the effects of acetylcholine, sodium nitroprusside and phenylephrine were evaluated on blood pressure, aortic rings and isolated and perfused mesenteric bed. The blood was obtained for plasma Interleukin-6 (IL-6), C-reactive protein (CRP) and lipid evaluation. The mesenteric vessels were obtained to evaluate superoxide production and nitric oxide synthase 3 (NOS-3) expression.ResultsLigature induced periodontitis reduced endothelium-dependent vasodilatation, a hallmark of endothelial dysfunction. This effect was associated with an increase in systemic inflammatory markers (IL-6 and CRP), worsens on lipid profile, increased vascular superoxide production and reduced NOS-3 expression. It is interesting to note that many of these effects were transitory.ConclusionPeriodontitis induced a transient systemic and vascular inflammation which leads to endothelial dysfunction, an initial step for cardiovascular diseases. Moreover, the animal model of periodontitis used here may represent a valuable tool for studying the relationship between periodontitis and endothelial dysfunction

Avaliação dos impactos socioeconômicos de projetos energéticos – eletrificação rural: área de concessão da ENERSUL – MS

This article evaluates the impacts of the Program of Rural Electrification called “Luz no Campo” on the energy profile of rural properties located in the state of Mato Grosso do Sul. The evaluation period comprehends three phases – ex-ante, intermediary and ex-post – of the field research and is based on the area of concession of the utility ENERSUL. It presents a panel of the local energy consumption and its relations with the family incomes. It also identifies some changes that occurred as a result of the access to electric energy. The results intend to help the decision makers in the formulation of public policies that aim at promoting the optimization of the use of energetic resources and atpromoting rural development.Key words: Energy, Rural electrification, Sustainable development.Este artigo busca avaliar os impactos no perfil energético das propriedades rurais, decorrentes da implementação do Programa de Eletrificação Rural “Luz no Campo” para a área de concessão da ENERSUL – Empresa Energética de Mato Grosso do Sul, durante três fases da pesquisa de campo: a fase ex-ante, a fase intermediária e a fase ex-post. Será apresentado um painel do perfil de consumo energético das comunidades e, além disso, as relações entre esse consumo e a renda familiar dessa população. Serão identificadas algumas transformações que o homem do campo sofre com a chegada da energia elétrica. Os resultados auxiliarão os tomadores de decisão na formulação de políticas públicas que promovam a otimização do uso de recursos energéticos, assim como na promoção do desenvolvimento rural.Palavras-chave: Energia, Eletrificação rural, Desenvolvimento sustentável

Avaliação dos impactos socioeconômicos de projetos energéticos – eletrificação rural: área de concessão da ENERSUL – MS

This article evaluates the impacts of the Program of Rural Electrification called “Luz no Campo” on the energy profile of rural properties located in the state of Mato Grosso do Sul. The evaluation period comprehends three phases – ex-ante, intermediary and ex-post – of the field research and is based on the area of concession of the utility ENERSUL. It presents a panel of the local energy consumption and its relations with the family incomes. It also identifies some changes that occurred as a result of the access to electric energy. The results intend to help the decision makers in the formulation of public policies that aim at promoting the optimization of the use of energetic resources and atpromoting rural development.Key words: Energy, Rural electrification, Sustainable development.Este artigo busca avaliar os impactos no perfil energético das propriedades rurais, decorrentes da implementação do Programa de Eletrificação Rural “Luz no Campo” para a área de concessão da ENERSUL – Empresa Energética de Mato Grosso do Sul, durante três fases da pesquisa de campo: a fase ex-ante, a fase intermediária e a fase ex-post. Será apresentado um painel do perfil de consumo energético das comunidades e, além disso, as relações entre esse consumo e a renda familiar dessa população. Serão identificadas algumas transformações que o homem do campo sofre com a chegada da energia elétrica. Os resultados auxiliarão os tomadores de decisão na formulação de políticas públicas que promovam a otimização do uso de recursos energéticos, assim como na promoção do desenvolvimento rural.Palavras-chave: Energia, Eletrificação rural, Desenvolvimento sustentável

Effect of synthesis temperature on crystallinity, morphology and cell viability of nanostructured hydroxyapatite via wet chemical precipitation method: Effect of temperature on hydroxyapatite properties

Hydroxyapatite (HA) is the main natural mineral constituent of bones and is a good alternative for biomedical applications because it is osteoconductive, non-allergenic, and non-carcinogenic, which ensures high biocompatibility. A commonly used method for obtaining hydroxyapatite is the wet route, which is simple and low-cost, produces only water as a final residue, and provides HA with a crystallinity comparable to that of bone tissue, which favors its biocompatibility. Therefore, the objective of this work is to synthesize hydroxyapatite via the wet chemical precipitation method at different temperatures (4°C, 30°C, 50°C, or 70°C) to observe the influence of temperature on crystallinity, morphology, and cytotoxicity. The results of X-ray diffraction show that all syntheses resulted in pure hydroxyapatite, while increasing the temperature led to higher crystallinity (10.6% to 56.2%) and the crystal size was slightly affected. The increase in temperature changed the particle shape from irregular to needle-like. Cell viability was tested by PicoGreen® in VERO cells for samples at concentrations of 30 and 300µg/mL, and the samples synthesized at 4°C, with lower crystallinity, caused less DNA damage to cells compared to the negative control. &nbsp

Strongyloides venezuelensis: efeito de antimicrobiano e imunossupressor no curso da infecção em camundongos da linhagem AKR/J

Groups of AKR/J strain of mice infected by Strongyloides venezuelensis and treated with Ceftazidima, Dexametasona or with both drugs concomitantly had been killed on 3rd, 7th and 12th day after infection and their lungs and intestines were processed for histological studies. In lungs of all infected groups, it was verified an inflammatory infiltrated (neutrophils and mononuclear cells) in the third day after the infection. In 7th and 12th day after the infection, the inflammatory reaction becomes reduced in control and antimicrobial treated groups, but not in immunosuppressed animals (with or without antimicrobial treatment). Analysis of the duodenal mucosa confirmed the presence of parasites in all groups of animals. On the 7th day after infection it was observed significant alterations in the small intestine of control (infected) and infected and treated with antimicrobial groups with presence of inflammatory foci, constituted by mononuclear and eosinophils in mucosa, associate to a large amount of parasites, mainly in the region of the epithelium and sub epithelium. Two others groups (infected and immunosuppressed mice with or without antimicrobial), did not present inflammatory process. Goblet cells were less evident in mucosa suggesting a reduction in mucous production. In the 12th day of the infection, the treated with antimicrobial and control groups presented a reduced number of parasites and a discrete inflammatory reaction in the small intestine while the immunosuppressed groups showed more parasites in duodenum. The permanence of the infection by S. venezuelensis in immunosuppressed animals was prolonged in relation to other groups, remaining until the 49th day after infection. The results indicate that the interference of treatments in the population of intestinal microbiota favours the parasitism by Strongyloides venezuelensis.Camundongos da linhagem AKR/J infectados pelo Strongyloides venezuelensis e tratados com Ceftazidima, Dexametasona ou com ambas as drogas foram sacrificados no terceiro, sétimo e décimo segundo dia após infecção. Pulmões e intestino delgado foram processados para histologia. Verificaram-se, nos pulmões dos quatro grupos infectados, infiltrados inflamatórios (neutrófilos e mononucleares) no terceiro dia após a infecção. No sétimo e décimo segundo dia após a infecção, o processo inflamatório se torna reduzido nos animais controle e tratados com antimicrobiano, mas não nos animais imunossuprimidos (com ou sem tratamento com antimicrobiano). A análise da mucosa duodenal confirmou a presença de parasitos em todos os grupos de animais. No 7º dia após a infecção, foram observadas alterações significativas no duodeno dos grupos controle (infectados) e dos infectados e tratados com antimicrobiano, com presença de infiltrado inflamatório, constituído de mononucleares e eosinófilos na mucosa, associada a uma maior quantidade de parasitos, principalmente na região do epitélio e sub-epitélio. Os outros dois grupos (camundongos infectados e imunossuprimidos com ou sem antimicrobiano), não apresentaram processo inflamatório. A mucosa apresentou células caliciformes menos evidentes, sugerindo uma redução de produção de muco. No décimo segundo dia da infecção, os grupos não tratados e tratados apenas com antimicrobiano apresentaram um número reduzido de parasitos e um discreto processo inflamatório no duodeno, enquanto os grupos tratados com imunossupressor (com ou sem antimicrobiano) mostraram presença de um maior número do parasito no duodeno. A permanência da infecção do S. venezuelensis foi mais prolongada nos animais imunossuprimidos em relação aos demais grupos, permanecendo até o 490dia após infecção. Os resultados indicam que a interferência dos tratamentos no equilíbrio da microbiota intestinal favoreceu o parasitismo pelo S. venezuelensis

Healing, Antioxidant and Cytoprotective Properties of Indigofera truxillensis in Different Models of Gastric Ulcer in Rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The present study evaluated the antiulcerogenic activity and mechanisms of the aqueous (AqF 100 mg/kg) and ethyl acetate (AcF 50 mg/kg) fractions from Indigofera truxillensis leaves. This dose was selected to assess its activity on ulcer healing and its action on gastric acid and mucus secretion, prostaglandin production and antioxidant enzyme activity (superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and glutathione reductase (GSH-Rd)). Gastric ulcer was induced by absolute ethanol. Antisecretory action, mucus and prostaglandin production, healing and antioxidant enzyme activities were evaluated for both fractions. AqF and AcF significantly inhibited the gastric mucosal damage caused by ethanol. This effect was statistically significant at 100 and 50 mg/kg compared with the vehicle. Neither fraction interfered with gastric secretion. AcF increased the PGE(2) production, and both fractions increased mucus production. L-NAME did not alter the gastroprotection exerted by the fractions, but N-ethylmaleimide attenuated only AcF. In the ischemia/reperfusion model both fractions inhibited the mucosal damage. AcF increased SOD, GSH-Px and GSH-Rd activity, but AqF increased only SOD and GSH-Px. In the acetic acid-induced ulcer model AcF only accelerated ulcer healing. These results showed that Indigofera truxillensis acted as a gastroprotective agent, stimulating protective factors and antioxidants enzymes.13111497314991Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Mechanisms of the Gastric Antiulcerogenic Activity of Anacardium humile St. Hil on Ethanol-Induced Acute Gastric Mucosal Injury in Rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Leaves and bark infusions Anacardium humile St. Hil. (Anacardiaceae), known as in Brazil as "cajuzinho do cerrado", have been used in folk medicine as an alternative treatment for ulcers and gastritis. This study evaluated the gastroprotective activity of an ethyl acetate extract of the leaves of A. humile (AcF) and the mechanism involved in this gastroprotection. Pretreatment concentrations (50, 100, 200 mg.kg(-1)) were administered by gavage. Following a 60 min. period, all the rats were orally administered 1 mL of absolute ethanol. One hour after the administration of ethanol, all groups were sacrificed, and the gastric ulcer index was calculated. Prostaglandin PGE(2) concentration, gastric adherent mucous, and the participation of nitric oxide (NO) and sulfhydryl compounds in the gastroprotection process were also analyzed using the most effective tested dose (50 mg.kg(-1)). A histological study of the glandular stomach for the evaluation of the epithelial damage and mucus content was also performed. AcF significantly reduced the gastric damage produced by ethanol. This effect was statistically significant for the 50 mg.kg(-1) group compared to control. Also, it significantly increased the PGE(2) (by 10-fold) and mucous production, while pretreatment with NG-nitro-L-arginine methyl ester (L-NAME) or N-ethylmaleimide (NEM) completely abolished the gastroprotection. AcF has a protective effect against ethanol, and this effect, might be due to the augmentation of the protective mechanisms of mucosa.151071537166Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq