Pre-treatment with IL-6 potentiates ß-cell death induced by pro-inflammatory cytokines.

BACKGROUND: Type I Diabetes mellitus (T1D) is characterized by a specific destruction of ß-cells by the immune system. During this process pro-inflammatory cytokines are released in the pancreatic islets and contribute for ß-cells demise. Cytokine-induced iNOS activation, via NF-κB, is implicated in induction of ß-cells death, which includes ER stress activation. Physical exercise has been used as an adjunct for better glycemic control in patients with T1D, since it is able to increase glucose uptake independent of insulin. Recently, it was observed that the release of IL-6 by skeletal muscle, during physical exercise, could prevent ß-cells death induced by pro-inflammatory cytokines. However, the molecular mechanisms involved in this beneficial effect on ß-cells are not yet completely elucidated. Our aim was to evaluate the effect of IL-6 on ß-cells exposed to pro-inflammatory cytokines. RESULTS: Pre-treatment with IL-6 sensitized INS-1E cells to cytokine-induced cell death, increasing cytokine-induced iNOS and Caspase-3 expression. Under these conditions, however, there was a decrease in cytokines-induced p-eIF2-α but not p-IRE1expression, proteins related to ER stress. To address if this prevention of adequate UPR response is involved in the increase in ß-cells death markers induced by IL-6 pre-treatment, we used a chemical chaperone (TUDCA), which improves ER folding capacity. Use of TUDCA increased cytokines-induced Caspase-3 expression and Bax/Bcl-2 ratio in the presence of IL-6 pre-treatment. However, there is no modulation of p-eIF2-α expression by TUDCA in this condition, with increase of CHOP expression. CONCLUSION: Treatment with IL-6 alone is not beneficial for ß-cells, leading to increased cell death markers and impaired UPR activation. In addition, TUDCA has not been able to restore ER homeostasis or improve ß-cells viability under this condition, suggesting that other mechanisms may be involved.

Fractions of Selaginella convoluta (Arn) Spring (Selaginellaceae) attenuate the nociceptive behavior events in mice / Frações de Selaginella convoluta (Ar) Spring (Selaginellaceae) atenuam a resposta nociceptiva em camundongos

Abstract Selaginella convoluta (Arn.) Spring is a species popularly known as "jericó", and used in folk medicine as analgesic and anti-inflammatory. This study aimed to investigate in mice the antinociceptive and anti-inflammatory activities of the hexane (Sc-Hex) and chloroform (Sc-CHCl3) fractions (100, 200 and 400 mg/kg) obtained by partition of crude ethanol extract from S. convoluta. The preliminary phytochemical analysis of the fractions was performed. Antinociceptive activity was evaluated by writhing, formalin and hot-plate tests. Anti-inflammatory activity was evaluated using carrageenan-induced pleurisy. The rota-rod test was used to evaluate motor coordination. Preliminary phytochemical screening showed that the Sc-Hex and the Sc-CHCl3 were positive for the presence of flavonoids, anthracene derivatives, quinones, triterpenes and steroids. Inhibition of writhing was observed for fractions tested. The Sc-Hex at all doses tested was effective in reducing the nociceptive behavior produced by formalin only in the second phase. However, the Sc-CHCl3 decreased the paw licking time in the first and second phases. In the hot plate no significant effect was observed for any fraction. In the rota-rod test, treated mice did not demonstrate any significant motor performance changes. In the carrageenan-induced pleurisy, Sc-CHCl3 (200 mg/kg) reduced cell migration to the pleural cavity. These results reveal the antinociceptive properties of S. convoluta , which support, in part, its traditional use, since the fractions did not presented significant activity in the inflammatory response profile. We further verify that this antinociceptive effect could be by activation of nociceptive peripheral pathway.

Resumo Selaginella convoluta é uma espécie popularmente conhecida como "jericó", e usada na medicina popular como analgésica e anti-inflamatória. Este estudo teve como objetivo investigar em camundongos as atividades antinociceptiva e anti-inflamatória das frações hexânica (Sc-Hex) e clorofórmica (Sc-CHCl3) (100, 200 e 400 mg/kg) obtidas por partição do extrato etanólico bruto de S. convoluta . A análise fitoquímica preliminar das frações foi realizada. A atividade antinociceptiva foi avaliada pelos testes de contorções abdominais, formalina e placa quente. A atividade anti-inflamatória foi avaliada usando pleurisia induzida por carragenina. O teste do rota-rod foi utilizado para avaliar a coordenação motora. A triagem fitoquímica preliminar mostrou que Sc-Hex e Sc-CHCl3 apresentaram reação positiva para a presença de flavonoides, derivados antracênicos, quinonas, triterpenos e esteroides. A inibição das contorções foi observada para as frações testadas. Sc-Hex em todas as doses testadas foi efetivo na redução do comportamento nociceptivo produzido pela formalina somente na segunda fase. No entanto, Sc-CHCl3 diminuiu o tempo de lambida da pata na primeira e segunda fases. Na placa quente, nenhum efeito significativo foi observado para qualquer fração. No teste do rota-rod os camundongos tratados não demonstraram mudanças significativas na coordenação motora. Na pleurisia induzida por carragenina, Sc-CHCl3 (200 mg/kg) reduziu a migração celular para a cavidade pleural. Estes resultados revelam a propriedade antinociceptiva de S. convoluta, justificando, em parte, seu uso tradicional, uma vez que os extratos não apresentaram atividade significativa no perfil de resposta inflamatória. Verificamos ainda que esse efeito antinociceptivo mostrou-se ligado à ativação da via periférica nociceptiva.

Fractions of Selaginella convoluta (Arn.) Spring (Selaginellaceae) attenuate the nociceptive behavior events in mice.

Selaginella convoluta (Arn.) Spring is a species popularly known as "jericó", and used in folk medicine as analgesic and anti-inflammatory. This study aimed to investigate in mice the antinociceptive and anti-inflammatory activities of the hexane (Sc-Hex) and chloroform (Sc-CHCl3) fractions (100, 200 and 400 mg/kg) obtained by partition of crude ethanol extract from S. convoluta. The preliminary phytochemical analysis of the fractions was performed. Antinociceptive activity was evaluated by writhing, formalin and hot-plate tests. Anti-inflammatory activity was evaluated using carrageenan-induced pleurisy. The rota-rod test was used to evaluate motor coordination. Preliminary phytochemical screening showed that the Sc-Hex and the Sc-CHCl3 were positive for the presence of flavonoids, anthracene derivatives, quinones, triterpenes and steroids. Inhibition of writhing was observed for fractions tested. The Sc-Hex at all doses tested was effective in reducing the nociceptive behavior produced by formalin only in the second phase. However, the Sc-CHCl3 decreased the paw licking time in the first and second phases. In the hot plate no significant effect was observed for any fraction. In the rota-rod test, treated mice did not demonstrate any significant motor performance changes. In the carrageenan-induced pleurisy, Sc-CHCl3 (200 mg/kg) reduced cell migration to the pleural cavity. These results reveal the antinociceptive properties of S. convoluta , which support, in part, its traditional use, since the fractions did not presented significant activity in the inflammatory response profile. We further verify that this antinociceptive effect could be by activation of nociceptive peripheral pathway.

Use of biopolymeric coating hydrophobized with beeswax in post-harvest conservation of guavas.

Biopolymeric coatings are effective in maintaining the chemical and sensory characteristics of fruits and vegetables because they preserve innumerable nutrients effectively for long periods of storage. Coatings based on cornstarch (3%), cassava starch (2%) and gelatin (5%) were synthesized. Beeswax (BW) was added as a hydrophobic agent in the polymeric network at different concentrations relative to the dry base of biopolymer. The biofilm containing 10% BW presented the best results in the water vapor transmission rate (WVTR) and the effect of the BW incorporation was also evaluated from surface roughness and SEM. An 80% increase in elasticity and 15% decrease in solubility indicated resistance against unfavorable environmental conditions. Physical-chemical analysis showed that its use minimized weight loss, guaranteeing adequate ripening of the fruits during 15 days in a regulated environment: 15 °C ±â€¯2 °C, 90% ±2% RH. The sensorial attributes indicated better acceptability of these fruits, demonstrating the potential of this coating.

Incorporation of bentonite clay in cassava starch films for the reduction of water vapor permeability.

Complete factorial planning 23 was applied to identify the influence of the cassava starch(A), glycerol(B) and modified clay(C) content on the water vapor permeability(WVP) of the cassava starch films with the addition of bentonite clay as a filler, its surface was modified by ion exchange from cetyltrimethyl ammonium bromide. The films were characterized by X-ray diffraction(XRD), fourier transform by infrared radiation(FTIR), atomic force microscopy(AFM) and scanning electron microscopy(SEM). The factorial analysis suggested a mathematical model thats predicting the optimal condition of the minimization of WVP. The influence of each individual factor and interaction in the WVP was investigated by Pareto graph, response surface and the optimization was established by the desirability function. The sequence of the degree of statistical significance of the investigated effects on the WVP observed in the Pareto graph was C>B>A>BC>AC. Interactions AB, BC and AC showed that the modified clay was the factor of greater significance.

Fertility restoration locus and cytoplasm types in onion.

The objective of this study was the identification of the cytoplasmic types and the genotyping for the fertility restoration nuclear locus (Ms) in 59 onion accessions, aiming at the selection of 'A' and 'B' lines essential for the obtainment of hybrids. Three markers were used to identify the cytoplasm 5' cob, orfA501, and orf725, and two were used for the Ms locus (AcSKP1 and AcPMS1). The two types of male-sterile cytoplasm ('S' and 'T'), as well as fertile cytoplasm ('N'), and the Ms and ms alleles in both homozygosity and heterozygosity were detected in the 59 genotypes evaluated in the experiment. The frequencies of the 5' cob/orfA501 and orf725 markers, as well as of the markers AcSKP1 and AcPMS1, were close in the onion accessions evaluated in this study. In the Brazilian germplasm, the frequencies of the 'N', 'S', and 'T' cytoplasm were approximately 0.47, 0.28, and 0.25, respectively, whereas the allele frequencies of Ms and ms were 0.52 and 0.48, respectively. The accessions Régia, EHCEB 20146, EHCEB 201427, Alvorada, Serrana, Crioula Mercosul, EHCEB 20142, BRS 367, Rainha, Juporanga, and Alfa SF C-XI have potential for the identification of 'A' and 'B' lines, since they presented mixtures of cytoplasm and different allele frequencies for Ms. All the plants of the accessions EHCEB 20142040/EHCEB 20141040, EHCEB 20142028/EHCEB 20141028, and EHCEB 20112006/EHCEB 20111006 were in the Nmsms and Smsms conditions, and have the potential for 'B' and 'A' lines, respectively, for the CMS-S system. All the plants of the accessions EHCEB 20142027/EHCEB 20141027, EHCEB 20102019/EHCEB 20101019, and Alfa SF 'B'/Alfa SF 'A' were in Nmsms and Tmsms conditions, and have the potential for 'B' and 'A' lines, respectively, for the CMS-T system.

Analysis of genetic variability of commercial melon cultivars using SSR molecular markers.

Taking into account the scenario of melon production (Cucumis melo L.) in Brazil, it is notable the importance of the culture in the fruit market. The preference of the national market is concentrated in the fruits of the yellow type. Therefore, it is possible that the genetic base of the commercial cultivars is relatively narrow due to the loss of important genes in the breeding process, and it is possible to introgress external genes for increasing recombination possibilities that generate sources of resistance to the main diseases and superior genotypes. This study evaluated the genetic variability of 87 melon accessions composed of 72 commercial cultivars belonging to the Inodorus and Cantaloupensis groups and 15 plant introductions (PIs). Forty-four polymorphic SSR primers were used to amplify a total of 202 alleles. The expected heterozygosity presented an average of 0.519 and the PIC index of 0.458, being moderately informative. The dendrogram generated for the 72 cultivars grouped the genotypes into 2 main groups, and there was no association with the classification of groups of the genotypes in the grouping. The number of SSR markers was sufficient to predict wide genetic variability among the cultivars studied, with the similarity between 0.35 and 0.98. The dendrogram presented for the 72 cultivars and the 15 PI genotypes was not associated with the classification of the genotypes in the grouping, and the 15 PIs were well dispersed with similarity indexes that resemble the two groups studied. A set of 25 primers was identified that were useful in distinguishing the 72 cultivars. These primers can be used in later research with the cultivars, as well as in crop protection situations, being an important tool in the useful and rapid distinction of genotypes, and in commercial disputes regarding the certification of the main melon cultivars used in the country.

Fractions of Selaginella convoluta (Arn.) Spring (Selaginellaceae) attenuate the nociceptive behavior events in mice

Abstract Selaginella convoluta (Arn.) Spring is a species popularly known as jericó, and used in folk medicine as analgesic and anti-inflammatory. This study aimed to investigate in mice the antinociceptive and anti-inflammatory activities of the hexane (Sc-Hex) and chloroform (Sc-CHCl3) fractions (100, 200 and 400 mg/kg) obtained by partition of crude ethanol extract from S. convoluta. The preliminary phytochemical analysis of the fractions was performed. Antinociceptive activity was evaluated by writhing, formalin and hot-plate tests. Anti-inflammatory activity was evaluated using carrageenan-induced pleurisy. The rota-rod test was used to evaluate motor coordination. Preliminary phytochemical screening showed that the Sc-Hex and the Sc-CHCl3 were positive for the presence of flavonoids, anthracene derivatives, quinones, triterpenes and steroids. Inhibition of writhing was observed for fractions tested. The Sc-Hex at all doses tested was effective in reducing the nociceptive behavior produced by formalin only in the second phase. However, the Sc-CHCl3 decreased the paw licking time in the first and second phases. In the hot plate no significant effect was observed for any fraction. In the rota-rod test, treated mice did not demonstrate any significant motor performance changes. In the carrageenan-induced pleurisy, Sc-CHCl3 (200 mg/kg) reduced cell migration to the pleural cavity. These results reveal the antinociceptive properties of S. convoluta , which support, in part, its traditional use, since the fractions did not presented significant activity in the inflammatory response profile. We further verify that this antinociceptive effect could be by activation of nociceptive peripheral pathway.

Resumo Selaginella convoluta é uma espécie popularmente conhecida como jericó, e usada na medicina popular como analgésica e anti-inflamatória. Este estudo teve como objetivo investigar em camundongos as atividades antinociceptiva e anti-inflamatória das frações hexânica (Sc-Hex) e clorofórmica (Sc-CHCl3) (100, 200 e 400 mg/kg) obtidas por partição do extrato etanólico bruto de S. convoluta . A análise fitoquímica preliminar das frações foi realizada. A atividade antinociceptiva foi avaliada pelos testes de contorções abdominais, formalina e placa quente. A atividade anti-inflamatória foi avaliada usando pleurisia induzida por carragenina. O teste do rota-rod foi utilizado para avaliar a coordenação motora. A triagem fitoquímica preliminar mostrou que Sc-Hex e Sc-CHCl3 apresentaram reação positiva para a presença de flavonoides, derivados antracênicos, quinonas, triterpenos e esteroides. A inibição das contorções foi observada para as frações testadas. Sc-Hex em todas as doses testadas foi efetivo na redução do comportamento nociceptivo produzido pela formalina somente na segunda fase. No entanto, Sc-CHCl3 diminuiu o tempo de lambida da pata na primeira e segunda fases. Na placa quente, nenhum efeito significativo foi observado para qualquer fração. No teste do rota-rod os camundongos tratados não demonstraram mudanças significativas na coordenação motora. Na pleurisia induzida por carragenina, Sc-CHCl3 (200 mg/kg) reduziu a migração celular para a cavidade pleural. Estes resultados revelam a propriedade antinociceptiva de S. convoluta, justificando, em parte, seu uso tradicional, uma vez que os extratos não apresentaram atividade significativa no perfil de resposta inflamatória. Verificamos ainda que esse efeito antinociceptivo mostrou-se ligado à ativação da via periférica nociceptiva.

Time-dependence of lung injury in mice acutely exposed to cylindrospermopsin.

Cylindrospermopsin is a cyanobacterial toxin of increasing environmental importance, as it can lead to disease if orally or intravenously absorbed. However, its in vivo lung impairment has not been documented. Thus, we aimed at verifying whether cylindrospermopsin can induce lung injury and establish its putative dependence on the time elapsed since exposure. BALB/c mice were intratracheally injected with either saline (NaCl 0.9%, 50 µL, SAL group, n = 12) or a sublethal dose (70 µg/kg) of semi-purified extract of cylindrospermopsin (CYN groups, n = 52). Lung mechanics, histological and biochemical analyses, and cylindrospermopsin presence in lungs and liver were determined in independent groups at 2, 8, 24, 48, and 96 h after cylindrospermopsin instillation. There was a significant increase in static elastance at 24 and 48 h after exposure to cylindrospermopsin, while viscoelastic component of elastance and viscoelastic pressure rose at 48 h. Alveolar collapse augmented in CYN groups at 8 h. A significant increase in polymorphonuclear influx into lung parenchyma, as well as a higher myeloperoxidase activity started off at 24 h. Exposure to cylindrospermopsin increased lipid peroxidation and superoxide dismutase activity and reduced catalase activity in CYN groups. The toxin was detected in lungs and liver of all CYN mice. In conclusion, cylindrospermopsin exposure impaired lung mechanics, which was preceded by lung parenchyma inflammation and oxidative stress.

Comparison of the genomes of two Xanthomonas pathogens with differing host specificities.

The genus Xanthomonas is a diverse and economically important group of bacterial phytopathogens, belonging to the gamma-subdivision of the Proteobacteria. Xanthomonas axonopodis pv. citri (Xac) causes citrus canker, which affects most commercial citrus cultivars, resulting in significant losses worldwide. Symptoms include canker lesions, leading to abscission of fruit and leaves and general tree decline. Xanthomonas campestris pv. campestris (Xcc) causes black rot, which affects crucifers such as Brassica and Arabidopsis. Symptoms include marginal leaf chlorosis and darkening of vascular tissue, accompanied by extensive wilting and necrosis. Xanthomonas campestris pv. campestris is grown commercially to produce the exopolysaccharide xanthan gum, which is used as a viscosifying and stabilizing agent in many industries. Here we report and compare the complete genome sequences of Xac and Xcc. Their distinct disease phenotypes and host ranges belie a high degree of similarity at the genomic level. More than 80% of genes are shared, and gene order is conserved along most of their respective chromosomes. We identified several groups of strain-specific genes, and on the basis of these groups we propose mechanisms that may explain the differing host specificities and pathogenic processes.

DNA repair-related genes in sugarcane expressed sequence tags (ESTs)

A identificaçäo e caracterizaçäo de genes envolvidos com reparo de DNA säo de grande interesse, dada a sua importância na manutençäo da integridade genômica. Além disso, a alta conservaçäo dos genes de reparo de DNA faz com que possam ser utilizados como fonte de informaçäo no que diz respeito à origem e evoluçäo das espécies. Os mecanismos relacionados à remoçäo de danos pelo reparo de DNA, bem como suas conseqüências biológicas, já säo bem conhecidas em bactérias, leveduras e animais. Entretanto, no que diz respeito a organismos vegetais, ainda há muito a ser investigado. No presente trabalho, apresentamos a identificaçäo dos genes envolvidos nas principais vias de reparo de DNA em cana-de-açúcar, através de uma análise de similaridade do banco de dados do projeto brasileiro Sugarcane Expressed Sequence Tag (SUCEST) com seqüências protéicas conhecidas disponíveis em outros bancos de dados públicos (National Center of Biotechnology Information (NCBI) e Munich Information Center for Protein Sequences (MIPS) Arabidopsis thaliana). Esta busca revelou que a gama de proteínas envolvidas no reparo de DNA em cana-de-açúcar é similar a de outros eucariotos. Mesmo assim, foi possível identificar algumas características interessantes encontradas apenas em vegetais, provavelmente em funçäo do seu processo evolutivo independente. As vias de reparo do DNA aqui representadas incluem fotorreativaçäo, reparo excisäo de bases, reparo excisäo de nucleotídeos, reparo mismatch, end-joinning näo homólogo, reparo por recombinaçäo homóloga e tolerância a lesões. Este trabalho descreve as principais diferenças encontradas na maquinaria de reparo de DNA de células vegetais em relaçäo àquela de organismos nos quais encontra-se bem descrita. Tais diferenças chamam a atençäo para um potencial de mecanismos distintos em vegetais, que merecem futuras investigações.

Schistosoma mansoni egg-induced hepatic granulomas in mice deficient for the interferon-gamma receptor have altered populations of macrophages, lymphocytes and connective tissue cells.

Systemic production and mobilization of inflammatory cells and formation of hepatic periovular granulomas were studied in Schistosoma mansoni-infected mice with deficient interferon gamma (IFN-gamma) receptor (IFN-gammaR(o/o)). The impaired IFN-gamma signaling did not cause a significant modification of the overall kinetics of inflammatory cells, but mutant mice developed smaller hepatic periovular granulomas with a two-fold reduction in all the cell lineages. In granulomas of normal mice, the fully differentiated macrophages were progressively predominant, whilst in IFN-gammaR(o/o) mice, the granulomas contained a higher percentage of immature and proliferating monocytes. Granulomas of IFN-gammaR(o/o) mice had an enhanced and accelerated fibrotic reaction, corresponding to an increased content of proliferative and activated connective tissue cells. Simultaneously, their granulomas had an increased ratio of T over B cells, with an increase in CD8(+) and a reduction in CD4(+) T cells. The functional IFN-gamma receptor was not required for initial recruitment of monocytes and lymphocytes into granulomas, but it was necessary for the maturation of macrophages, upregulation of major histocompatibility class 2 (MHC-II) expression and consequent stimulation of lymphocyte subpopulations depending upon the MHC-II-mediated antigen presentation.

The "lazy S-shaped" plication of the SMAS-platysma musculoaponeurotic system: a 10-year review.

Morphological changes that appear with the aging process are expressed by loss of the youthful face and neck contours. A "lazy S-shaped" plication involving the superficial musculoaponeurotic system (SMAS)-Platysma has been performed to recover the face and neck contour. On the neck, the posterior segment of the lateral portion of the muscle was stretched upward and backward to the mastoid. On the face, plication was done laterally and superiorly to the corner of the mouth toward the zygomatic arch. The anterior platysmal bands were approached in the midline to reinforce the muscle sling of the neck. Extensive skin dissection with suction lipectomy under direct visualization of the submental and cervical fat were associated. Photographs were taken preoperatively, 1 year, and 10 years after the surgery. Face and neck contour at 10 years was compared with that at 1 year after the surgery and with the patient look before the surgery. Improvement of the face and neck contour was achieved in all patients at 1 year postoperatively. At 10 years after the surgery the face and neck contour was equal or eventually better than before the surgery.

H-Shaped, double-contour plication in abdominoplasty.

H-shaped, double-contour plication, by reducing the longitudinal and transverse abdominal diameters, improves the waistline and reinforces the entire anterior abdominal wall, with better accommodation of the cutaneous flap. The decrease in the longitudinal diameter allows lowering and fixation of the cutaneous flap without tension.

Mice lacking the gamma interferon receptor have an impaired granulomatous reaction to Schistosoma mansoni infection.

The egg-induced granulomatous reaction in Schistosoma mansoni-infected individuals develops within the portal system of the liver and is the major pathological finding in schistosomiasis. We have infected mice lacking the gamma interferon (IFN-gamma) receptor with S. mansoni larvae and studied the development of hepatic granulomas in these mutant mice in comparison to that in control wild-type mice. In the absence of IFN-gamma activity, a dramatic reduction in the size and architecture of the granuloma was observed. Granulomas from mutant mice were smaller than those from the control group and showed a significant reduction in the number of infiltrating inflammatory cells. Moreover, they appear to prematurely progress to the chronic phase of the reaction at a time when the control group still has acute inflammation. Our data suggests a pivotal role for IFN-gamma in the early events of the granulomatous reaction in vivo.