Characterization of a beta-1,3-glucanase active in the alkaline midgut of Spodoptera frugiperda larvae and its relation to beta-glucan-binding proteins

Spodoptera frugiperda beta-1,3-glucanase (SLam) was purified from larval midgut. It has a molecular mass of 37.5 kDa, an alkaline optimum pH of 9.0, is active against beta-1,3-glucan (laminarin), but cannot hydrolyze yeast beta-1,3-1,6-glucan or other polysaccharides. The enzyme is an endoglucanase with low processivity (0.4), and is not inhibited by high concentrations of substrate. In contrast to other digestive beta-1,3-glucanases from insects, SLam is unable to lyse Saccharomyces cerevisae cells. The cDNA encoding SLam was cloned and sequenced, showing that the protein belongs to glycosyl hydrolase family 16 as other insect glucanases and glucan-binding proteins. Multiple sequence alignment of beta-1,3-glucanases and beta-glucan-binding protein supports the assumption that the beta-1,3-glucanase gene duplicated in the ancestor of mollusks and arthropods. One copy originated the derived beta-1,3-glucanases by the loss of an extended N-terminal region and the beta-glucan-binding proteins by the loss of the catalytic residues. SLam homology modeling suggests that E228 may affect the ionization of the catalytic residues, thus displacing the enzyme pH optimum. SLam antiserum reacts with a single protein in the insect midgut. Immunocytolocalization shows that the enzyme is present in secretory vesicles and glycocalyx from columnar cells. (C) 2010 Elsevier Ltd. All rights reserved.

Novel electroluminescent devices containing Eu(3+)-(2-acyl-1,3-indandionate) complexes with TPPO ligand

Fabrication and electroluminescent properties of devices containing europium complexes of general formula [Eu(ACIND)(3)(TPPO)(2)], where ACIND, 2-acyl-1,3-indandionate ligands: and TPPO, triphenylphosphine oxide. as emitter layers are discussed. The double-layer devices based on these complexes present the following configurations: device 1: ITO/TPD/[Eu(AlND)(3)(TPPO)(2)]/Al: device 2: ITO/TPD/[Eu(ISOV-IND)(3)(TPPO)(2)]/Al and device 3: ITO/TPD/[Eu(BIND)(3)(TPPO)(2)]/Al, where AlND, 2-acetyl-1,3-indandionate; ISOVIND, 2-isovaleryl-1,3-indandionate; and BIND, 2-benzoyl-1,3-indandionate, respectively. These devices exhibited photo and electroluminescent emissions. An important characteristic presented by devices is that their electroluminescent (EL) spectra, in the region of (5)D(0) -> (7)F(J) (J = 0, 1, 2, 3 and 4) transitions of Eu(3+) ion, show profiles that are different from photoluminescent (PL) ones. In addition to narrow bands arising from intraconfigurational-4f(6) transitions, devices 1 and 2 also exhibited a broad band with maximum at around 500 nm which is assigned to electrophosphorescence from the indandionate ligands. On the other hand, EL spectra of device 3 present only narrow bands from (5)D(0) -> (7)F(J) transitions. [Eu(ACIND)(3)(TPPO)(2)] complexes are promising candidates to prepare efficient organic light-emitting devices (OLEDs) when compared with those containing Eu(3+)-complexes of aliphatic beta-diketonate anions. (C) 2009 Elsevier B.V. All rights reserved.

Palladium-catalyzed oxidative homocoupling of potassium alkenyltrifluoroborates: synthesis of symmetrical 1,3-dienes

Herein, we describe a convenient method for the synthesis of symmetrical 1,3-dienes employing an oxidative palladium-catalyzed homocoupling of potassium alkenyltrifluoroborates providing products in good yields relative to existing methodologies. This is the first report of a cross-dimerization of potassium alkenyltrifluoroborates. (C) 2009 Elsevier Ltd. All rights reserved.

Síntese e caracterização de novos materiais líquido-cristalinos derivados de bifenilcarboxilatos e 3,5-isoxazolinas

Este trabalho mostra a síntese de novos materiais poliméricos utilizando como unidade mesogênica anéis bifenilas, com um grupo espaçador de onze unidades metilênicas, com uma cadeia polimérica acrilato e que apresentam como grupo terminal uma porção quiral sintetizada a partir de L-isoleucina. Os monômeros apresentaram comportamento enantiotrópico mesofases esmética C e A quiral. Do mesmo modo, os polímeros apresentaram mesofase esmética A estável no aquecimento e no resfriamento. Foi realizado também a síntese de poliacrilatos tendo presente como grupo de ligação na unidade mesogênica o anel isoxazolina. Para a construção do heterocíclico de cinco membros utilizou-se como metodologia de síntese a reação de cicloadição 1,3-Dipolar. Foram sintetizados materiais poliméricos e não poliméricos. Os monômeros isoxazolínicos não apresentaram mesofase, entretanto, os homopolímeros apresentaram mesofase nemática. As Isoxazolinas não-poliméricas não apresentaram mesofase. A síntese de isoxazoliltolanos foi realizada combinando a metodologia de cicloadição 1,3-dipolar e a reação de Sonogashira. Foram sintetizados compostos que apresentaram mesofase nemática monotrópica.

Produção e purificação de β-1,3 glicanases em Metarhizium anisopliae

Metarhizium anisopliae é um fungo cosmopolita com capacidade de infectar uma grande variedade de hospedeiros, estando entre eles o carrapato Boophilus microplus. A penetração de M. anisopliae em seus hospedeiros ocorre de forma ativa onde a cutícula constitui a principal barreira. A penetração é um processo multifatorial, porém, o emprego de pressão mecânica e a secreção de enzimas hidrolíticas parecem ser fundamentais para o seu sucesso. M. anisopliae, quando cultivado em meios com fontes de carbono que mimetizam a cutícula de seus hospedeiros, secreta enzimas como proteases, quitinases e lipases. Atualmente, o emprego de técnicas que identificam genes diferencialmente expressos (RDA) mostrou o possível envolvimento de outras enzimas, como as β-glicanases, durante o processo de penetração. A descoberta da ocorrência de modificações morfológicas como espessamento e perda da definição da parede celular nas extremidades das hifas que penetram na cutícula do carrapato sustentam ainda mais o possível envolvimento de enzimas que degradam as β-glicanas nas etapas iniciais da infecção. Neste trabalho, foi investigada a produção de β-1,3- glicanases pela linhagem E6 de M. anisopliae como também, buscou-se purificar as enzimas produzidas. A síntese e secreção de β-1,3-glicanases foram verificadas em meio contendo diferentes fontes de carbono sendo a secreção diferenciada dependendo da condição testada. A utilização de glicose em determinadas concentrações pareceu inibir a secreção enzimática. Duas das condições testadas, N-acetilglicosamina (NAG) 0,5% e parede celular de Rizoctonia solani 0,5%, foram utilizadas para a produção enzimática em larga escala. O sobrenadante dos cultivos em fermentador foi submetido ao processo de purificação que constou de três etapas: concentração por ultrafiltração com membrana de celulose regenerada, aplicação em coluna de troca iônica QSepharose Fast Flow e aplicação em coluna de filtração em gel Superdex 75. O emprego deste protocolo permitiu a purificação parcial de uma β-1,3-glicanase com aproximadamente 95kDa, secretada durante a fermentação em presença de parede celular de Rizoctonia solani, e de outra, com aparentemente a mesma massa molecular secretada em fermentação utilizando NAG 0,5% como fonte de carbono. Durante este trabalho, também foi confirmada a presença de pelo menos um gene que codifica uma exo-β-1,3-glicanase no genoma da linhagem E6 de M. anisopliae. Por fim, o estudo das β-1,3 glicanases em M. anisopliae é justificado pela importância destas enzimas em variados aspectos do desenvolvimento do fungo bem como, pelo seu possível envolvimento na infecção de hospedeiros.

Botryosphaeran, a new substrate for the production of beta-1,3-glucanases by Botryosphaeria rhodina and Trichoderma harzianum Rifai

Botryosphaeria rhodina and Trichoderma harzianum Rifai were grown on botryosphaeran (an exopolysaccharide (EPS) of the beta-1,3; 1,6-D-Glucan type produced by B. rhodina) as sole carbon source with the objective of producing beta-glucanases of the beta-type. Conditions for beta-1,3-glucanase production by T harzianum were examined by a statistical response surface method, and showed maximal enzyme production at 5 days growth in media containing 1.5 g/1 of EPS. Good agreement was obtained between the experimental values of beta-1, 3-glucanase activity and the corresponding values predicted by the mathernatical model. The crude beta-1,3-glucanase preparations were active towards a number of different beta-1,3-glucans and beta-glucosides. The mycelium of B. rhodina also proved to be a good substrate for beta-1,3-glucanase production by both fungal species. (c) 2005 Published by Elsevier Ltd.

Oxidation of melatonin and its catabolites, N-1-acetyl-N (2)-formyl-5-methoxykynuramine and N-1-acetyl-5-methoxykynuramine, by activated leukocytes

N-1-acetyl-N-2-formyl-5-methoxykynuramine (AFMK) and N-1-acetyl-5-methoxykynuramine (AMK), two melatonin catabolites, have been described as potent antioxidants. We aimed to follow the kinetics of AFMK and AMK formation when melatonin is oxidized by phorbol myristate acetate (PMA) and lipopolysaccharide (LPS)-activated leukocytes. An HPLC-based method was used for AFMK and AMK determination in neutrophil and peripheral blood mononuclear cell cultures supernatants. Samples were separated isocratically on a C18 reverse-phase column using acetonitrile/H2O (25:75) as the mobile phase. AFMK was detected by fluorescence (excitation 340 nm and emission 460 nm) and AMK by UV-VIS absorbance (254 nm). Activation of neutrophils and mononuclear cells with PMA produces larger amounts of AFMK than activation with LPS, probably due to the lower levels of reactive oxygen species formation and myeloperoxidase (MPO) degranulation that occurs when cells are stimulated with LPS. The concentration of AMK found in the supernatant was about 5-10% (from 18-hr cultures) compared with AFMK. This result may reflect its reactivity. Indeed AMK, but not AFMK, is easily oxidized by activated neutrophils in a MPO and hydrogen peroxide-dependent reaction. In conclusion, we defined a simple procedure for the determination of AFMK and AMK in biological samples and demonstrated the capacity of leukocytes to oxidize melatonin and AMK.

Avaliação da desadaptação das selas de próteses parciais removíveis após 1 a 5 anos de uso

The non-adaptation of the removable partial prosthesis (RPP) base to fibromucosal tissue is caused by resorption of residual ridges (RRR). The onset of bone resorption, which occurs after tooth extraction and continues throughout life, is accelerated by local or systemic factors. Aim: Assess the degree of non-adaptation of removable partial prosthesis saddles and the factors that influence it. Methodology: A sectional study was conducted with 81 patients using RPP who had their prostheses installed between 2003 and 2007 (1 to 5 years of use) at the Faculty of Dentistry of the Universidade Federal do Rio Grande do Norte (UFRN). After anamnese and clinical examination, a cast was made with polyether-based material, using the base of the prosthesis to make the impression. The base of the saddle was loaded with the casting material and positioned in the mouth, applying pressure on the supports. After polymerization, the material was removed from the saddle and measurements were taken at 3 different points using a pachymeter. Results: The non-adaptation of the saddle increased significantly with years of use (p = 0.005). The tooth-tissue supported prostheses obtained higher mean non-adaptation values than those of tooth supported prostheses (p < 0.001). Flaccid mucosa showed the worst non-adaptation results, which were statistically different from resilient mucosa (p < 0.001). The greater the extension of the saddle, the greater the non-adaptation (p < 0.001). The natural tooth antagonistic arch yielded better results than did RPP and total prosthesis (p < 0.001). Saddle non-adaptation at the free end was less near the pillar tooth and greater in the more posterior region (p < 0.001). When adaptation of the supports to the niches was poor, greater saddle non-adaptation occurred than when it was good or fair (p < 0.001). Saddles located in the posterior region of the arch had greater non-adaptation than those in the anterior region (p = 0.023). Conclusion: The mean non-adaptation of the saddle to the residual ridges was 0.27 mm. It can be concluded that, even with the use of RPP, bone height reduction was slight within the 1-5-year period of use. The following are factors that influence adaptation of the RPP saddle base: years of use, age, force transmission path to the alveolar bone, location of the toothless area, antagonistic arch, type of mucosa, adaptation of supports to the niche and extension of the saddle

Avaliação imuno-histoquímica das galectinas -1, -3 e -7 em displasia epitelial ora

Introdução: A displasia epitelial oral (DEO) é uma lesão potencialmente maligna, cujo diagnóstico e gradação se baseia na histologia das alterações arquiteturais e citológicas, preconizados pela OMS, que divide a lesão em leve, moderada e severa, o qual é subjetivo. Maior concordância é observada no uso do sistema binário (baixo/alto risco), o qual está relacionado ao risco de transformação maligna. As galectinas constituem uma família de lectinas e estão envolvidas na tumorigênese, sendo a -1, -3 e -7 as mais investigadas, devido a expressão alterada em cânceres orais. Materiais e métodos: Foi analisada a expressão imuno-histoquímica dessas proteínas em 50 espécimes de DEO (21 baixo/ 29 alto risco) e 5 de mucosa oral normal e relacionamos com a presença/ausência de marcação, padrão de distribuição, intensidade, localização epitelial (estratificação) (1/3 inferior, médio e superior), e localização celular (compartimento) (núcleo, citoplasma e membrana) . Resultados: Dos 29 casos de alto e dos 21 de baixo risco, 21 (72,4%) e 12 (57,1%) foram positivos para a galectina -1, respectivamente. Dessa forma, de 50 casos, 33 foram positivos. O núcleo e citoplasma foram positivos em 91,7% nas de baixo risco e em 90,5% nas de alto. Todos os casos de mucosa normal foram negativos. Com relação a galectina -3, dos 21 casos das DEOs de baixo risco, 12 (57,1%) apresentaram expressão e dos 29 casos das DEOs de alto risco, 15 (51,7%) foram positivos, havendo imunoexpressão em um total de 27 casos. O padrão difuso, assim como a fraca intensidade foram os mais freqüentes para os 2 graus. O núcleo e o citoplasma foram a localização mais comum tanto nas lesões de baixo (58,3%), quanto nas de alto risco (66,7%). Quatro casos de mucosa normal foram positivos, com marcação membranar e intensidade fraca. Dos 21 casos das DEOs de baixo risco, 17 (81%) apresentaram expressão imuno-histoquímica para a galectina -7 e das 29 DEOs de alto risco, 27 (93,1%) foram positivos. Então, a expressão imuno-histoquímica da galectina -7 foi observada em 44 casos, a maioria com intensidade de moderada a forte. O núcleo e o citoplasma foram a localização mais freqüente, nas de baixo (70,6%) e alto risco (66,7%). Quatro espécimes de mucosa normal marcaram membrana em terço médio e superior, com intensidade moderada a forte. Conclusões: Alterações na expressão das galectinas -3 e -7 e principalmente da -1 sugerem seu envolvimento na fisiopatologia das displasias, participando do processo de transformação de fenótipo normal para o displásico.

Study of copper complexes adsorbed on a silica gel surface chemically modified with 2-amino-1,3,4-thiadiazole

The isotherms of adsorption of CuX2 (X=Cl-, Br-, ClO4-) by silica gel chemically modified with 2-amino-1,3,4-thiadiazole were studied in acetone and ethanol solutions: at 298 K. The following equilibria constants (in 1 mol(-1)) were determined: (a) CuCl2: 3.5 x 10(3) (ac), 2.0 x 10(3) (eth); (b) CuBr2: 2.8 x 10(3) (ac), 2.0 x 10(3) (eth); (c) Cu(ClO4)(2): 1.8 x 10(3) (ac), 1.0 x 10(3) (eth); ac = acetone, eth = ethanol. The electron spin resonance spectra of the surface complexes indicated a tetragonal distorted structure in the case of lower degrees of metal loading on the chemically modified surface. The d-d electronic transition spectra showed that for the ClO4-, complex, the peak of absorption did not change for any degree of metal loading, and for Cl- and Br- complexes, the peak maxima shifted to a higher energy region with a lower metal loading. (C) 1998 Elsevier B.V. B.V. All rights reserved.

Purification and properties of acid phosphatase (EC 3.1.3.2) secreted by strain 74A of the mould Neurospora crassa

Both P-i-repressible acid phosphatases, IIb (mycelial) and IIc (extracellular), synthesized by Neurospora crassa and purified to apparent homogeneity by 7.5% PAGE, are monomers, are inhibited by 2 mM ZnCl2 and are nonspecifically stimulated by salts. However, the IIc form is activated by p-nitrophenylphosphate (in a negative cooperativity effect with a K-0.5 of 2.5 mM) whereas form IIb shows Michaelis kinetics, with a K-m of 0.5 mM. Thus, since both enzymatic forms may be expressed by the same gene (pho-3), it is possible that post-translational modifications lead to the excretion of an enzymatic form with altered Michaelis kinetics compared with the enzymatic form retained by the mycelium.

The 3.4-3.5 Ga Sao Jose do Campestre massif, NE Brazil: remnants of the oldest crust in South America

The oldest fragment of continental crust recognized in South America occurs as an isolated Archean enclave in northeastem Brazil's Borborema Province, ca. 600 Ma Brasiliano-Pan African orogenic belt. This Archean fragment, the Sao Jose do Campestre massif, is surrounded by large tracts of 2.2-2.0 Ga Paleoproterozoic gneisses and is located more than 600-1500 km from the much larger assemblages of Archean rocks found in the Sao Fransciso and Amazonian cratons, located to the south and west, respectively. Geochronological studies of the Sao Jose do Campestre massif show that its oldest rocks contain zircons with U-Pb ages up to 3.5 Ga and Sm-Nd T-DM model ages of more than 3.7 Ga, indicating that they represent reworked crust. This older nucleus is flanked by both reworked and juvenile 3.25 and 3.18 Ga rocks which arc intruded by both 3.00 and 2.69 Ga plutonic bodies. The protracted evolution the Sao Jose do Campestre massif is consistent with that of a larger continental mass as opposed to a small crustal fragment that grew in isolation. As such, the Sao Jose do Campestre massif is interpreted as representing a detached piece of an evolved craton that became entrained with younger rocks during a subsequent Paleoproterozoic accretionary-orogenic event. This hypothesis is bolstered by the presence of Paleoproterozoic gneisses that envelop the Sao Jose do Campestre massif, as well as the existence of ca. 2.0 Ga metamorphic zircon and monazite within its rocks. The occurrence of several different Archean cratonic basement inliers within the greater Paleoproterozoic crustal framework of the Borborema Province suggests that cratonic slices spalled off one or more larger Archean masses prior to the ca. 2.2-2.0 Ga Paleoproterozoic orogenic collage. A important challenge is to link these older fragments to their parent cratons. Although results are not unique, the pattern of ages and isotopic signatures observed in the Sao Jose do Campestre massif is similar to that seen in parts of the Sao Francisco Craton, and it is possible that the Sao Jose do Campestre massif is a fragment of an Archean continental fragment formed during an episode of continental breakup prior to 2200 Ma. (C) 2003 Elsevier B.V. All rights reserved.

Crystal structure of potassium trifluoro(2-methyl-1,3-dithiano)borate, K(C5H9BF3S2)

C5H9BF3KS2, triclinic, P (1) over bar (no. 2), a = 11.9238(5) angstrom, b = 13.6060(5) angstrom, c = 14.0280(3) angstrom, alpha = 114.995(2)degrees, beta = 92.035(2)degrees, gamma = 92.390(2)degrees, V = 2057.4 angstrom(3), Z = 8, R-gt(F) = 0.049, wR(ref)(F-2) = 0.117, T = 296 K.

THE PHO-2A MUTANT OF NEUROSPORA-CRASSA WHICH IS DEFICIENT IN PI-REPRESSIBLE ALKALINE-PHOSPHATASE (EC 3.1.3.1) IS ALSO DEFECTIVE IN PI-REPRESSIBLE ACID-PHOSPHATASE (EC 3.1.3.2)

1. The mycelial Pi-repressible acid phosphatase presented p-nitrophenylphosphatase activity with negative cooperativity and Michaelian behavior when synthesized by the wild-type and pho-2A mutant strains of Neurospora crassa, respectively.2. The major acid phosphatase present in cell extracts of the pho-2A mutant of N. crassa grown in low Pi medium is more thermolabile (t1/2 = 4 min at 54-degrees-C, pH 5.4) than that of the wild strain (stable for at least 80 min at 54-degrees-C, pH 5.4).3. The pho-2A mutant of N. crassa secreted a more thermolabile acid phosphatase (t1/2 = 30 min at 50-degrees-C, pH 5.4) than the wild strain (t1/2 of at least 80 min at 50-degrees-C, pH 5.4).4. The pho-2A mutant of N. crassa synthesized a more thermolabile acid phosphatase (t1/2 = 37 min at 54-degrees-C, pH 5.4) than the wild strain in high Pi medium (t1/2 = 14 min al 54-degrees-C, pH 5.4).5. The pleiotropic nature of the pho-2 locus and its possible involvement in the mechanism of phosphatase secretion by N. crassa are proposed.

Electrolytic treatment of wastewater containing n-phenyl-n '-1,3-dimethylbutyl-p-phenylenediamine

This study investigated the effects of electrolytic treatment using Dimensionally Stable Anode (DSA, 70%TiO2/30%RuO2) type electrodes in simulated wastewater containing aromatic amine n-phenyl-n'-1,3-dimethylbutyl-p-phenylenediamine (Flexzone 7P). A low direct current density of 0.025 A cm(-2) was applied for periods up to 60 minutes and a 52.6% decrease in Flexzone 7P concentration was observed. Ultraviolet-visible spectra, gas chromatography, toxicity and biodegradation tests were carried out with the aim of verifying the toxic by-products that were formed. Ultraviolet-visible spectra of simulated wastewater exhibited changes in the aromatic amine's molecular structure. Additionally, based on the S. cerevisiae toxicity test, it was observed that detoxification of the wastewater occurred after 15 minutes of electrolysis. It was also observed that five minutes of treatment were sufficient to improve the biodegradation rate, determined through the respirometric Bartha method.