Efeito do exercício físico sobre a liberação de interleucina-1[beta], interleucina-6 e fator de necrose tumoral-[alfa] em homens portadores de doença pulmonar obstrutiva crônica

A atividade física intensa pode induzir resposta inflamatória subclínica e aumento nos níveis plasmáticos de citocinas pró-inflamatórias. O objetivo deste estudo foi avaliar a relação entre a liberação de citocinas (IL-1β, IL-6, e TNF-α), o exercício físico agudo e o exercício regular em pacientes com doença pulmonar obstrutiva crônica (DPOC). Foram estudados 18 pacientes do sexo masculino com DPOC moderada a muito grave, divididos em dois grupos: 11 pacientes foram incluídos em programa de reabilitação pulmonar (RP) durante 8 semanas e 7 pacientes sem atividade física regular foram incluídos como grupo controle (C). Todos os pacientes realizaram espirometria, teste de exercício cardiopulmonar incremental máximo e teste de endurance em cicloergômetro com carga constante (60% da carga máxima do teste incremental) no início do projeto e após oito semanas. Foi coletado sangue venoso periférico para dosagem de citocinas, antes e 15 minutos após os testes de endurance (TE1 e TE2). IL-1β, IL-6, e TNF-α foram dosadas com kits ELISA específicos (Quantikine®, R&D Systems). Os pacientes submetidos à RP liberaram menos IL-1β que os controles após o treinamento (RP: TE1 0,96±0,66; TE2 -0,24±0,27 pg/ml; grupo C: TE1 -1,48±1,14; TE2 0,66±0,61 pg/ml; p=0,03). Não houve diferença significativa na liberação de IL-6 quando comparados os dois testes de endurance (RP: TE1 0,44±1,21; TE2 0,80±1,24 pg/ml; grupo C: TE1 0,88±0,85; TE2 0,78±0,95 pg/ml; p=0,68). Não foi observada diferença na liberação de IL-6 entre os dois grupos. Apenas cinco pacientes (quatro no grupo da RP) liberaram TNF-α e o exercício não modificou o seu padrão de liberação (RP: TE1 2,86±1,18; TE2 2,57±1,37pg/ml; grupo C: TE1 4,98; TE2 6,84 pg/ml; p=0,14). Não houve associação significativa entre intensidade de exercício e liberação de citocinas (IL-1β r=0,10; IL-6 r=-0,23). Houve maior liberação de IL-6 após o TE2 nos pacientes que apresentaram exacerbação da DPOC (exacerbados 9,59±1,32; estáveis 6,31±0,92 pg/ml; p=0,03) e não houve diferença nos níveis de IL-1β. Apenas pacientes com exacerbação da DPOC liberaram TNF-α (2,82±1,48 pg/ml). Concluiu-se que o exercício físico regular reduz a liberação de IL-1β e as exacerbações estimulam a liberação de IL-6 e TNF-α em pacientes com DPOC.

Purification and structural characterisation of (1 -> 3 ; 1 -> 6)-beta-D-glucans (botryosphaerans) from Botryosphaeria rhodina grown on sucrose and fructose as carbon sources: a comparative study

Two botryosphaerans, exopolysaccharides (EPS) secreted by the ascomyceteous fungus Botryosphaeria rhodina, when grown on sucrose and fructose as sole carbon sources, were structurally compared after their isolation from the culture medium. Both EPS were submitted to trypsin digestion, and eluted as a single peak on gel filtration. Total acid hydrolysis yielded only glucose, and data from methylation analysis and Smith degradation indicated that both EPS constituted a main chain of glucopyranosyl beta(1 -> 3) linkages substituted at O-6. The products obtained after partial acid hydrolysis demonstrated side chains consisting of glucosyl- and gentiobiosyl- linked beta(1 -> 6) residues. C-13-NMR spectroscopy studies showed that all glucosidic linkages were of the beta-configuration. The carbon source affected the side chain structures of botryosphaeran but not the main chain makeup. Sucrose produced less branching (21%) than fructose (31%). (c) 2005 Published by Elsevier Ltd.

Structural characterization of Botryosphaeran: a (1 -> 3;1 -> 6)-beta-D-glucan produced by the ascomyceteous fungus, Botryosphaeria sp.

The exopolysaccharide, Botryosphaeran, produced by the ligninolytic, ascomycetcous fungus Botryosphaeria sp., was isolated from the extracellular fluid by precipitation with ethanol, and purified by gel permeation chromatography to yield a carbohydrate-rich fraction (96%) composed mainly of glucose (98%). Infra-red and C-13 NMR spectroscopy showed that all the glucosidic linkages were in the beta-configuration. Data from methylation analysis and Smith degradation indicated that Botryosphaeran was a (1 --> 3)-beta-(D)-glucan with approx 22% side branching at C-6. The products obtained from partial acid hydrolysis demonstrated that the side branches consisted of single (1 --> 6)-beta-linked glucosyl, and (1 --> 6)-beta-linked gentiobiosyl residues.[GRAPHICS](C) 2003 Elsevier Ltd. All rights reserved.

Chemical Modification of Botryosphaeran: Structural Characterization and Anticoagulant Activity of a Water-Soluble Sulfonated (1 -> 3)(1 -> 6)-beta-D-Glucan

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Amniotic Fluid Interleukin-1 Beta and Interleukin-6, but not Interleukin-8 Correlate with Microbial Invasion of the Amniotic Cavity in Preterm Labor

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

MKK3/6-p38 MAPK signaling is required for IL-1 beta and TNF-alpha-induced RANKL expression in bone marrow stromal cells

Coupled bone turnover is directed by the expression of receptor-activated NF-kappa B ligand (RANKL) and its decoy receptor, osteoprotegerin (OPG). Proinflammatory cytokines, such as interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) induce RANKL expression in bone marrow stromal cells. Here, we report that IL-1 beta and TNF-alpha-induced RANKL requires p38 mitogen-activating protein kinase (MAPK) pathway activation for maximal expression. Real-time PCR was used to assess the p38 contribution toward IL-1 beta and TNF-alpha-induced RANKL mRNA expression. Steady-state RANKL RNA levels were increased approximately 17-fold by IL-1 beta treatment and subsequently reduced similar to 70%-90% when p38 MAPK was inhibited with SB203580. RANKL mRNA stability data indicated that p38 MAPK did not alter the rate of mRNA decay in IL-1 beta-induced cells. Using a RANKL-luciferase cell line receptor containing a 120-kB segment of the 5' flanking region of the RANKL gene, reporter expression was stimulated 4-5-fold by IL-1 beta or TNF-alpha treatment. IL-1 beta-induced RANKL reporter expression was completely blocked with specific p38 inhibitors as well as dominant negative mutant constructs of MAPK kinase-3 and -6. In addition, blocking p38 signaling in bone marrow stromal cells partially inhibited IL-1 beta and TNF-alpha-induced osteoclastogenesis in vitro. Results from these studies indicate that p38 MAPK is a major signaling pathway involved in IL-1 beta and TNF-alpha-induced RANKL expression in bone marrow stromal cells.

Interleukin-1 beta and Interleukin-6 Expression and Gene Polymorphisms in Subjects with Peri-Implant Disease

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Evaluation of the beta-glucanolytic enzyme complex of Trichoderma harzianum Rifai for the production of gluco-oligosaccharide fragments by enzymatic hydrolysis of 1,3;1,6-beta-D-glucans

Botryosphaeran, a new exopolysaccharide from the endophytic fungus Botryosphaeria rhodina MAMB-05, and algal laminarin were hydrolyzed by partially-fractionated enzymes of the beta-glucanolytic complex from Trichoderma harzianum Rifai. beta-Glucanase fractions (F-I and F-II) separated by gel permeation chromatography presented different modes of attack on botryosphaeran and laminarin. Botryosphaeran was hydrolyzed to the extent of 66% (F-I) and 98% (F-II) within 30 min, and its main hydrolysis products were gluco-oligosaccharides of DP >= 4, with lesser amounts of glucose, di- and tri-saccharides. The action of enzyme fractions I and II on laminarin resulted in 15% conversion to glucose, while the percentage of saccharification was radically different (70% for F-I and 25% for F-II). The different product arrays within the polysaccharide hydrolysates can be explained by the difference in the enzymes' specificities within each enzyme fraction, and the molecular structures of the polysaccharides and their complexity.

Three exopolysaccharides of the beta-(1 -> 6)-D-glucan type and a beta-(1 -> 3;1 -> 6)-D-glucan produced by strains of Botryosphaeria rhodina isolated from rotting tropical fruit

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Triple helix conformation of botryosphaeran, a (1 -> 3;1 -> 6)-beta-D-glucan produced by Botryosphaeria rhodina MAMB-05

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

MKK3/6-p38 MAPK negatively regulates murine MMP-13 gene expression induced by IL-1 beta and TNF-alpha in immortalized periodontal ligament fibroblasts

Matrix metalloprotease-13 (MMP-13) or collagenase-3 is involved in a number of pathologic processes such as tumor metastasis and angiogenesis, osteoarthritis, rheumatoid arthritis and periodontal diseases. These conditions are associated with extensive degradation of both connective tissue and bone. This report examines gene regulation mechanisms and signal transduction pathways involved in Mmp-13 expression induced by proinflammatory cytokines in periodontal ligament (PDL) fibroblasts. Mmp-13 mRNA expression was increased 10.7 and 9.5 fold after stimulation with IL-1 beta (5 ng/mL) and TNF-alpha (10 ng/mL), respectively. However, inhibition of p38 MAPKinase with SB203580 resulted in significant (p < 0.001) induction (23.2 and 18.1 fold, respectively) of Mmp-13 mRNA as assessed by real time PCR. Negative regulation of IL-1 induced Mmp-13 expression was confirmed by inhibiting p38 MAPK gene expression with siRNA. Transient transfection of dominant negative forms of MKK3 and MKK6 also resulted in increased levels of Mmp-13 mRNA after IL-1 beta stimulation. Mmp-13 mRNA expression induced by TNF-alpha was decreased by JNK and ERK inhibition. Western blot and zymogram analysis indicated that Mmp-13 protein expression induced by the proinflammatory cytokines were also upregulated by inhibition of p38 MAPK. Reporter gene experiments using stable cell lines harboring 660-bp sequence of the murine Mmp-13 proximal promoter indicated that transcriptional mechanisms were at least partially involved in this negative regulation of Mmp-13 expression by p38 MAPK and upstream MKK3/6. These results suggest a negative transcriptional regulatory mechanism mediated by p38 MAPK and upstream MKK3/6 on Mmp-13 expression induced by proinflammatory cytokines in PDL fibroblasts. (c) 2005 Elsevier B.V./International Society of Matrix Biology. All rights reserved.

An unusual C-6-C-6 '' linked flavonoid from Miconia cabucu (Melastomataceae)

Chromatographic fractionation of the methanolic extract from the leaves of Miconia cabucu Hoehne (Melastomataceae) afforded the first example of a C-6-C-6 linked flavone dimer, 5-hydroxy-4',7-dimethoxyflavone-(6-C-6)-5-hydroxy-3 4',7-trimethoxyflavone as well as the known compounds, quercetin- 3-O-alpha-L- rhamnopyranosyl-(2 -> 1)-O-beta-D-xylopyranoside, quercetin-3-O-alpha-L-rhamnopyranoside, myricetin-3-O-alpha-L-rhamnopyranoside, quercetin-3-O-beta-D-glucopyranoside, kaempferol -3-O-beta-D-(6-coumaroyl) -glucopyranoside and gallic acid. Their chemical identities were established by application of NMR spectroscopic methods including 2D-NMR, as well as UV and ESI-MS analyses. (c) 2007 Elsevier Ltd. All rights reserved.

Purification and characterization of two beta-glucosidases from the thermophilic fungus Thermoascus aurantiacus

beta-Glucosidase from the fungus Thermoascus aurantiacus grown oil semi-solid fermentation medium (using ground corncob as substrate) was partially purified in 5 steps - ultrafiltration, ethanol precipitation, gel filtration and 2 anion exchange chromatography runs, and characterized. After the first anion exchange chromatography, beta-glucosidase activity was eluted in 3 peaks (Gl-1, Gl-2, Gl-3). Only the Gl-2 and Gl-3 fractions were adsorbed on the gel matrix. Gl-2 and Gl-3 exhibited optimum pH at 4.5 and 4.0, respectively. The temperature optimum of both glucosidases was at 75-80 degreesC. The pH stability of Gl-2 (4.0-9.0) was higher than Gl-3 (5.5-8.5); both enzyme activities showed similar patterns of thermostability. Under conditions of denaturing gel chromatography the molar mass of Gl-2 and Gl-3 was 175 and 157 kDa, respectively. Using 4-nitrophenyl beta-D-glucopyranoside as substrate, K-m, values of 1.17 +/- 0.35 and 1.38 +/- 0.86 mmol/L were determined for Gl-2 and Gl-3, respectively. Both enzymes were inhibited by Ag+ and stimulated by Ca2+.

Estresse oxidativo em pacientes beta talassêmicos heterozigotos e com deficiência de glicose-6-fosfato desidrogenase

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)