Actinobacillus actinomycetemcomitans lipopolysaccharide induces interleukin-6 expression through multiple mitogen-activated protein kinase pathways in periodontal ligament fibroblasts

Actinobacillus actinomycetemcomitans plays a major role in the pathogenesis of aggressive periodontitis. Lipopolysaccharide (LPS) derived from A. actinomycetemcomitans is a key factor in inflammatory cytokine generation within periodontal tissues. In this study, we identify major mitogen-activated protein kinase (MAPK) signaling pathways induced by A. actinomycetemcomitans LPS, Escherichia coli LPS and interleukin-1 beta (IL-1 beta) in a murine periodontal ligament (mPDL) fibroblast cell line. Immunoblot analysis was used to assess the phosphorylated forms of p38, extracellular-regulated kinase (ERK) and c-jun N-terminal kinase (JNK) MAPK following stimulation with A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta. IL-6 mRNA induction was detected via reverse transcription-polymerase chain reaction, while protein levels were quantified via enzyme-linked immunosorbent assays (ELISA). We utilized biochemical inhibitors of p38, ERK and JNK MAPK to identify the MAPK signaling pathways needed for IL-6 expression. Additional use of stable mPDL cell lines containing dominant negative mutant constructs of MAPK kinase-3 and -6 (MKK-3/6) and p38 null mutant mouse embryonic fibroblast (MEF) cells were used to substantiate the biochemical inhibitor data. Blocking p38 MAPK with SB203580 reduced the induction of IL-6 mRNA by A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta by > 70%, > 95% and similar to 60%, respectively. IL-6 ELISA indicated that blocking p38 MAPK reduced the IL-6 protein levels induced by A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta by similar to 60%, similar to 50% and similar to 70%, respectively. All MAPK inhibitors significantly reduced the IL-6 protein levels induced by A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta whereas only p38 inhibitors consistently reduced the A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta induction of IL-6 mRNA steady-state levels. The contribution of p38 MAPK LPS-induced IL-6 expression was confirmed using MKK-3/6 dominant negative stable mPDL cell lines. Wild-type and p38 alpha(-/-) MEF cells provided additional evidence to support the role of p38 alpha MAPK in A. actinomycetemcomitans LPS-stimulated IL-6. Our results indicate that induction of IL-6 by E. coli LPS, IL-1 beta and A. actinomycetemcomitans LPS requires signaling through MKK-3-p38 alpha ERK, JNK and p38 MAPK in mPDL cells.

Avaliação química e da atividade antidiarréica das folhas de Byrsonima cinera DC. (Malpighiaceae)

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

Constituintes químicos das flores de Pterogyne nitens (Caesalpinioideae)

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

Toxicity of Dimorphandra mollis to Workers of Apis mellifera

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

New crystal forms of Diocleinae lectins in the presence of different dimannosides

Studying the interactions between lectins and sugars is important in order to explain the differences observed in the biological activities presented by the highly similar proteins of the Diocleinae subtribe. Here, the crystallization and preliminary X--ray data of Canavalia gladiata lectin (CGL) and C. maritima lectin (CML) complexed with Man(alpha 1-2)Man(alpha 1)OMe, Man(alpha 1-3)Man(alpha 1)OMe and Man(alpha 1-4)Man(alpha 1)OMe in two crystal forms [the complexes with Man(alpha 1-3)Man(alpha 1)OMe and Man(alpha 1-4)Man(alpha 1)OMe crystallized in space group P3(2) and those with Man(alpha 1-2)Man(alpha 1)OMe crystallized in space group I222], which differed from those of the native proteins (P2(1)2(1)2 for CML and C222 for CGL), are reported. The crystal complexes of ConA-like lectins with Man(alpha 1-4)Man(alpha 1)OMe are reported here for the first time.

Antifungal iridoids from the stems of Tocoyena formosa

A new antifungal iridoid compound, together with three active known iridoids, galioside, galioside aglucone, and apodanthoside, as well as 3-O-beta-D-glucopyranosyl-28-O-beta-D-glucopyranosyl quinovic acid and quercetin-3-O-methyl-3-O-beta-D-rhamnopyranoside, were isolated from the stems of Tocoyena formosa. This new compound was characterized by spectral data as 11-O-trans-feruloylteucrein.

Chemical composition of flavonoids and styrylpyrones and the genetic variability of isozymes in natural populations of Cryptocarya mandioccana Meisner (Lauraceae)

The chemical composition of leaves of 57 trees of Cryptocarya mandioccana from three populations of southeastern Brazil was investigated through HPLC, assaying six flavonoids, seven styrylpyrones, and seven unidentified compounds. From 51 of the former trees, genotypes were obtained from 40 polymorphic loci of 19 isozymes. Cluster analyses of the phytochemical and genetical variation revealed that trees exhibited four chemotypes and five clusters from isozymes, respectively. Discriminant analyses from selected variables of the isozymic and chemical data sets were performed, respectively, in relation to the four chemotypes and the five isozyme clusters. The classification of individuals presented respective error estimates of 9.16% and 13.57%, indicating that the genetic data could explain the clusters from chernotypes and vice versa at acceptable error levels. Linear regressions with Dummy variable showed significant association of locus Skdh-2 with quercetin-3-O-beta-D-glucopyranoside and cryptofolione, indicating that its alleles would be responsible for the chemotype variation between individuals. (c) 2006 Elsevier Ltd. All rights reserved.

Real-time monitoring and kinetic parameter estimation of the affinity interaction of jArtinM and rArtinM with peroxidase glycoprotein by the electrogravimetric technique

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

New biflavonoid and other flavonoids from the leaves of Chimarrhis turbinata and their antioxidant activities

A new biflavonol, named chimarrhoside (1), and eight known flavonol glycosides (2-9), were isolated from the leaves of Chimarrhis turbinata. Their structures were established on the basis of 1D and 2D NMR experiments as quercetin-3-O-rutinoside (2), kaempferol-3-O-rutinoside (3), kaempferol-3-O-α-L-rhamnopyranosyl-(1→6)-β-D-galactopyranoside (4), quercetin-3-O-α-L-rhamnopyranosyl-(1→6)-β-D- galactopyranoside (5), 6-hydroxy-rutin (6), kaempferol-3-O-D-galactopyranoside (7), kaempferol-3-O-D-glucopyranoside (8) and kaempferol-3-O-α- Lrhamnopyranosyl-(1→6)-α-L-rhamnopyranosyl-(1→4) -β-D-glucopyranoside (9). In addition, catechin (10) and catechin-(4α→8)-catechin-procyanidin B-3) (11) were isolated. The crude extract, fractions and isolated compounds were evaluated for their antioxidative properties using an autographic assay based on β-carotene bleaching on TLC plates, and spectrophotometric detection by reduction of the stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical. Flavonoids 2, 5, 6, 10 and 11 displayed strong free radical scavenging activity, when compared with the standards BHT and rutin. ©2005 Sociedade Brasileira de Química.

Annona sp: Plants with multiple applications as alternative medicine - a review

Annona species have been used as a natural remedy for a variety of illnesses with antiparasitic, antispasmodic, antidiarrheal, antiulcer, sedative, analgesic, hypotensive, and vermifugal effects. These properties are due to the presence of a number of bioactive compounds on the leaves, fruit, seeds, and stem. The aim of this review is to show the main species of Annona, their medicinal properties and the chemical constituents that may be related to these effects. In the leaves it is possible to find acetogenins, annopentocins A, B, and C, cis- and trans-annomuricin-D-ones, goniothalamicin, arianacin, and javoricin, related to anticancer properties. Quercetin-3-O-glucoside, also found in the leaves mediates antidiabetic and antioxidative effects. In the fruit are found annonaine, nornuciferine and asimilobine, associated to antidepressive effects. In the seeds are found muricatetrocin A and muricatetrocin B, longifolicin, corossolin, corossolone, uvarigrandin A, bullatacin, squamotatin. These acetogenins are associated with anticancer effects. Cyclosquamosin B, quercetin, and cyclosquamosin from the seeds have respectively vasorelaxant, antithyroidal and, antiinflammatory activity. In the stem parts there are several components as N-trans-feruloyltyramine, N-p-coumaroyltyramine, and N-trans-caffeoyltyramine, lignans, syringaresinol, syringaldehyde, beta-sitosterol and beta-sitosterol-beta-D-glucoside which exhibit antiplatelet aggregation activity. Copaene, patchoulane, 1H-cycloprop (e) azulene and kaur-16-en-19-oic acid found in the barks exhibit significant central as well as peripheral analgesic and antiinflammatory activities. The properties of the biological compounds in Annona species support information that may provide validation for its medicinal uses, but further studies should be performed to establish ideal and safe doses of consumption to ensure the effectiveness of the benefits. © 2012 Bentham Science Publishers.

Free radical scavenging activity of Kielmeyera variabilis (Clusiaceae)

As part of our ongoing research on antioxidant agents from Brazilian flora, we screened the free radical scavenging activity of two extracts and eight fractions of Kielmeyera variabilis (Clusiaceae) using DPPH (2,2-diphenyl-1- picrylhydrazyl-hydrate) and ABTS+ [2,2'-azinobis(3- ethylenebenzothiazoline-6-sulfonic acid)] colorimetric assays. The ethyl acetate and n-butanol fractions of the leaves of K. variabilis displayed the strongest activity (IC50 of 3.5 ± 0.3 and 4.4 ± 0.2 μg mL -1 for DPPH and 6.6 ± 0.4 and 3.1 ± 0.1 μg mL -1 for ABTS+, respectively). Chromatographic fractionation of the most potent fractions led to identification of three flavonols with previously described antioxidant activity, quercitrin (1), quercetin-3-O- glucoside (3), and quercetin-3-O-galactoside (4), and of one biflavone, podocarpusflavone A (2). This is the first time that the presence of these flavonoids in Kielmeyera variabilis has been reported. © 2013 by the authors.

Chromatic characteristics and color-related phenolic composition of Brazilian young red wines made from the hybrid grape cultivar BRS Violeta (BRS Rúbea×IAC 1398-21)

The color characteristics and the phenolic composition related to color of young red wines elaborated with the hybrid grape cultivar BRS Violeta, developed for its adaptation to sub-tropical climates in Brazil, have been studied. These wines are characterized with a deep red-purplish color, reaching color intensity averaging 24 units. In spite of being young red wines elaborated with short maceration time, their content in total polyphenols was very high (around 3692. mg/L, as gallic acid equivalents), especially when compared to similar Vitis vinifera young red wines. Within polyphenols, anthocyanins predominated (around 2037. mg/L, as malvidin 3,5-diglucoside equivalents) and they were almost exclusively anthocyanidin 3,5-glucosides (ca. 97%), mainly built from B-ring tri-substituted anthocyanidins (delphinidin. >. petunidin. ≈. malvidin) and having high proportion of p-coumaroylated derivatives (ca. 28%) that confer higher stability. The content of hydroxycinnamic acid derivatives was also remarkable (ca. 95. mg/L, as caffeic acid equivalents) and unknown glucose derivatives of p-coumaric acid accounted for ca. 42% of total HCAD. Finally, these found flavonols were mainly based on myricetin whereas kaempferol derivatives were missing, their total content being within the ranges usually found for V. vinifera wines, but reaching their top values (ca. 91. mg/L, as quercetin 3-glucoside equivalents). All the aforementioned data suggest that Violeta wine could be considered an important dietary source of healthy polyphenols with a moderate alcoholic content (ca. 11.6%). © 2013 Elsevier Ltd.

Phenolic composition of the berry parts of hybrid grape cultivar BRS Violeta (BRS Rubea×IAC 1398-21) using HPLC-DAD-ESI-MS/MS

The grape is considered a major source of phenolic compounds when compared to other fruits and vegetables, however, there are many cultivars with distinct characteristics directly linked to phenolic profile. Thus, the present study aimed to identify and quantify, for the first time and in detail, the phenolic compounds present in the skin, flesh and seeds of BRS Violeta grape berry using combination of SPE methodologies and analytical HPLC-DAD-ESI-MS/MS. The study was extended to the different berry parts and the most important grape and wine phenolic families, and has revealed interesting features. Violeta grape has a very thick skin (46% of grape weight) that accumulated the most of grape phenolic compounds: great amount of anthocyanins (3930. mg/kg, as malvidin 3,5-diglucoside), together with also important amounts of flavonols (150. mg/kg, as quercetin 3-glucoside), hydroxycinnamic acid derivatives (HCAD; 120. mg/kg, as caftaric acid), and proanthocyanidins (670. mg/kg, as (+)-catechin); in contrast, it seems to be a low resveratrol producer. Violeta grape seeds accounted for similar proportions of low molecular weight flavan-3-ols (mainly monomers; 345. mg/kg, as (+)-catechin) and proanthocyanidins (480. mg/kg, as (+)-catechin). Violeta grape is a teinturier cultivar, but it only contained traces of anthocyanins and low amounts of all the other phenolic types in its red-colored flesh. The anthocyanin composition of Violeta grape was dominated by anthocyanidin 3,5-diglucosides (90%). Within flavonols, myricetin-type predominated and kaempferol-type was missing. In addition to expected hydroxycinnamoyl-tartaric acids, several isomeric esters of caffeic and p-coumaric acids with hexoses were tentatively identified, accounting for relevant proportions within the pool of HCAD. Although pending of further confirmation over successive vintages, the aforementioned results suggest that BRS Violeta grape cultivar could be considered an interesting candidate for the elaboration of highly colored and antioxidant-rich grape juices and wines. © 2013 Elsevier Ltd.

Avaliação da atividade mutagênica e fitoestrogênica das plantas Davilla nitida Vahl. e Davilla elliptica St. Hil (Dilleniaceae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Desenvolvimento de filtros solares a partir de complexos metálicos de Zn2+ e/ou Ce3+ com quercetina para protetores solares

Pós-graduação em Química - IQ