Structural insights for fatty acid binding in a Lys49-phospholipase A(2): crystal structure of myotoxin II from Bothrops molojeni complexed with stearic acid

The crystal structure of dimeric Lys49-phospholipase A2 myotoxin-II from Bothrops moojeni (MjTX-II) co-crystallized with stearic acid (C18H36O2) has been determined at a resolution of 1.8 angstrom. The electron density maps permitted the unambiguous inclusion of six stearic acid molecules in the refinement. Two stearic acid molecules could be located in the substrate-binding cleft of each monomer in positions, which favor the interaction of their carboxyl groups with active site residues. The way of binding of stearic acids to this Lys49-PLA(2)s is analogous to phospholipids and transition state analogues to catalytically active PLA(2)s. Two additional stearic acid molecules were located at the dimer interface region, defining a hitherto unidentified acyl-binding site on the protein surface. The strictly conserved Lys122 for Lys49-PLA(2)s may play a fundamental role for stabilization of legend-protein complex. The comparison of MjTX-II/satiric acid complex with other Lys-PLA(2)s structures whose putative fatty acids were located at their active site is also analysed. Molecular details of the stearic acid/protein interactions provide insights to binding in croup I/II PLA(2)s and to the possible interactions of Lys49-PLA(2)s with target membranes. (c) 2004 Elsevier SAS. All rights reserved.

Identity by descent and candidate gene mapping of Richieri-Costa and Pereira syndrome

The Richieri-Costa-Pereira syndrome is a rare autosomal recessive disorder characterized by short stature, Robin sequence, cleft mandible, pre/postaxial anomalies and clubfoot. of 15 families reported with this disorder 14 are from Brazil suggesting a founder effect. We studied 15 families using identity-by-descent as a hypothesis to attempt gene localization We have examined through linkage analysis 497 polymorphicmarkers and also performed direct sequencing of exons for 10 candidate genes selected on the basis of their expression in the developing mandible and limb. No evidence for allele sharing at any locus tested or mutations in candidate genes was found. Additional higher resolution mapping, new families and other candidate genes might improve future chances of gene identification. (C) 2003 Wiley-Liss, Inc.

Dynamics of glyphosate-induced conformational changes of Mycobacterium tuberculosis 5-enolpyruvylshikimate-3-phosphate synthase (EC 2.5.1.19) determined by hydrogen-deuterium exchange and electrospray mass spectrometry

The enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) catalyzes the reaction between shikimate 3-phosphate and phosphoenolpyruvate to form 5-enolpyruvylshikimate 3-phosphate, an intermediate in the shikimate pathway, which leads to the biosynthesis of aromatic amino acids. EPSPS exists in an open conformation in the absence of substrates and/or inhibitors and in a closed conformation when bound to the substrate and/or inhibitor. In the present report, the H/D exchange properties of EPSPS from Mycobacterium tuberculosis (Mt) were investigated for both enzyme conformations using ESI mass spectrometry and circular dichroism (CD). When the conformational changes identified by H/D exchanges were mapped on the 3-D structure, it was observed that the apoenzyme underwent extensive conformational changes due to glyphosate complexation, characterized by an increase in the content of alpha-helices from 40% to 57%, while the beta-sheet content decreased from 30% to 23%. These results indicate that the enzyme underwent a series of rearrangements of its secondary structure that were accompanied by a large decrease in solvent access to many different regions of the protein. This was attributed to the compaction of 71% of alpha-helices and 57% of beta-sheets as a consequence of glyphosate binding to the enzyme. Apparently, MtEPSPS undergoes a series of inhibitor-induced conformational changes, which seem to have caused synergistic effects in preventing solvent access to the core of molecule, especially in the cleft region. This may be part of the mechanism of inhibition of the enzyme, which is required to prevent the hydration of the substrate binding site and also to induce the cleft closure to avoid entrance of the substrates.

Hydrogen/deuterium exchange mass spectrometry for characterizing phosphoenolpyruvate-induced structural transitions in Mycobacterium tuberculosis 5-enolpyruvylshikimate-3-phosphate synthase (EC 2.5.1.1)

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

The prevalence of human papillomavirus in the oropharynx in healthy individuals in a Brazilian population

Oncogenic human papillomavirus (HPV), a causative agent of uterine cervical cancer, has also been detected in head and neck squamous cell cancers, especially in squamous cell carcinomas of the tonsils. However, the true HPV prevalence in normal and neoplasic oropharyngeal mucosa remains uncertain. To determine the prevalence of HPV DNA in normal oropharyngeal mucosa of cancer-free individuals, a study was carried out on 50 Brazilian subjects. PCR was performed to identify HPV DNA in samples from four sites in the oropharynx (tonsils, soft palate, base of the tongue, and back wall of the pharynx). For amplification of the HPV DNA, MY09/11 consensus primerswere used, and specific genotypes were identified by dot-blot hybridization or cloning and sequencing. HPV DNA was present in 14.0% of the individuals, and the identified genotypes were 16, 18, 52, and 61. All these types are considered high-risk (HR) HPV. The tonsils and the soft palate were the sites with the highest HPV prevalence. This study shows the prevalence of HR HPV in the oropharynx of normal individuals. However, the prevalence of HPV is still unclear, and if HPV infection in a healthy it is not known individual predisposes to HPV-associated disease such as oropharyngeal cancer. Thus, it is important to assess the prevalence of HPV in cancer-free individuals, in order to compare it with the HPV prevalence in oropharyngeal carcinomas and to attempt to determine the true role of HPV in the development of head and neck squamous cell cancers. (c) 2006 Wiley-Liss, Inc.

Correlation of temperature induced conformation change with optimum catalytic activity in the recombinant G/11 xylanase A from Bacillus subtilis strain 168 (1A1)

The 1.7 angstrom resolution crystal structure of recombinant family G/11 beta-1,4-xylanase (rXynA) from Bacillus subtilis 1A1 shows a jellyroll fold in which two curved P-sheets form the active-site and substrate-binding cleft. The onset of thermal denaturation of rXynA occurs at 328 K, in excellent agreement with the optimum catalytic temperature. Molecular dynamics simulations at temperatures of 298-328 K demonstrate that below the optimum temperature the thumb loop and palm domain adopt a closed conformation. However, at 328 K these two domains separate facilitating substrate access to the active-site pocket, thereby accounting for the optimum catalytic temperature of the rXynA. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Amino acid sequence and crystal structure of BaP1, a metalloproteinase from Bothrops asper snake venom that exerts multiple tissue-damaging activities

BaP1 is a 22.7-kD P-I-type zinc-dependent metalloproteinase isolated from the venom of the snake Bothrops asper, a medically relevant species in Central America. This enzyme exerts multiple tissue-damaging activities, including hemorrhage, myonecrosis, dermonecrosis, blistering, and edema. BaP1 is a single chain of 202 amino acids that shows highest sequence identity with metalloproteinases isolated front the venoms of snakes of the subfamily Crotalinae. It has six Cys residues involved in three disulfide bridges (Cys 117-Cys 197, Cys 159-Cys 181, Cys 157-Cys 164). It has the consensus sequence H(142)E(143)XXH(146)XXGXXH(152), as well as the sequence C164I165M166, which characterize the metzincin superfamily of metalloproteinases. The active-site cleft separates a major subdomain (residues 1-152), comprising four a-helices and a five-stranded beta-sheet, from the minor subdomain, which is formed by a single a-helix and several loops. The catalytic zinc ion is coordinated by the N-epsilon2 nitrogen atoms of His 142, His 146, and His 152, in addition to a solvent water molecule, which in turn is bound to Glu 143. Several conserved residues contribute to the formation of the hydrophobic pocket, and Met 166 serves as a hydrophobic base for the active-site groups. Sequence and structural comparisons of hemorrhagic and nonhemorrhagic P-I metalloproteinases from snake venoms revealed differences in several regions. In particular, the loop comprising residues 153 to 176 has marked structural differences between metalloproteinases with very different hemorrhagic activities. Because this region lies in close proximity to the active-site microenvironment, it may influence the interaction of these enzymes with physiologically relevant substrates in the extracellular matrix.

Structural basis for branching-enzyme activity of glycoside hydrolase family 57: Structure and stability studies of a novel branching enzyme from the hyperthermophilic archaeon Thermococcus Kodakaraensis KOD1

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

Inflorescence and floral development of Dahlstedtia species (Leguminosae: Papilionoideae: Millettieae)

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

Oral Exfoliative Cytology for the Diagnosis of Paracoccidioidomycosis in a Patient with Human Immunodeficiency Virus A Case Report

BackgroundParacoccidioidomycosis is not the most common fungal disease in patients infected with human immunodeficiency virus (HIV), except for endemic regions in Latin America countriesCaseA 33-year-old man with HIV presented with mulberry-like lesions on the palate The diagnosis was made by exfoliative cytology and Papanicolaou staining Microscopic analysis revealed fungal structures with birefringent walls and exosporulation conferring an airplane radial motor appearance, or even bowel-like or goblet-like forms compatible with Paracoccidioides brasiliensisConclusionThis process spares the immunosuppressed patient from under going invasive biopsy procedures (Acta Cytol 2010,54 1127-1129)

Mucus extravasation and retention phenomena: a 24-year study

Background: Mucoceles are benign lesions related to the minor salivary glands and their respective ducts frequently affecting oral structures which are generally asymptomatic. Mucoceles are generally characterized by swollen nodular lesions preferentially located on the lower lip and differ from the so-called ranulas, which are lesions located on the floor of the mouth and related to the sublingual or submandibular glands.Methods: The objective of the present study was to analyze data such as age, gender, race and site of the lesion of 173 mucocele cases diagnosed at the Discipline of Stomatology, Sao Jose dos Campos Dental School, UNESP, over a period of 24 years (April 1980 to February 2003).Results: of the 173 cases analyzed, 104 (60.12%) were females and 69 (39.88%) were males. Age ranged from 4 to 70 years (mean +/- SD: 17 +/- 9.53) and most patients were in the second decade of life (n = 86, 49.42%); white (n = 124, 71.68%). The lower lip was the site most frequently affected by the lesions (n = 135, 78.03%), whereas the lowest prevalence was observed for the soft palate, buccal mucosa, and lingual frenum.Conclusion: In this study, mucoceles predominated in white female subjects in the second decade of life, with the lower lip being the most frequently affected site.

A new Sphagesaurus (Mesoeucrocodylia: Notosuchia) from the Upper Cretaceous of Monte Alto City (Bauru Group, Brazil), and a revision of the Sphagesauridae

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

PRODUÇÃO DE MUDAS DE VIDEIRA 'RUBI' PELO MÉTODO DE ENXERTIA DE MESA em ESTACAS HERBÁCEAS DOS PORTA-ENXERTOS IAC 313 'TROPICAL' E IAC 766 'CAMPINAS'

Com o objetivo de estudar o método de enxertia de mesa em estacas herbáceas para a obtenção de mudas de videira, experimentos foram desenvolvidos em câmara de nebulização, localizada no ripado do Departamento de Produção Vegetal da Faculdade de Ciências Agrárias e Veterinárias da UNESP, no período de novembro de 1997 a dezembro de 1999. Foram utilizadas estacas herbáceas dos porta-enxertos IAC 313 'Tropical' e IAC 766 'Campinas' e borbulhas e garfos da cultivar Rubi. Foram testados os métodos de enxertia de garfagem em fenda cheia e de borbulhia em placa, em estacas herbáceas sem raízes e enraizadas, o tipo de proteção do enxerto, o tamanho das estacas e o recipiente de enraizamento. Foram avaliados o enraizamento e o pegamento de enxertos. O método de enxertia de mesa com garfagem e com proteção do enxerto com saco plástico apresentou maior pegamento de enxerto aos 30 (45,25%) e 60 dias (66,20%) após a enxertia. O tamanho da estaca não afetou o pegamento dos enxertos. Contudo, em todos os experimentos, o índice de sobrevivência dos enxertos foi muito baixo ou nulo. Estacas sem enxertia proporcionaram maior porcentagem de enraizamento (65,62%) em relação às estacas enxertadas. O método de enxertia de mesa em estacas herbáceas dos porta-enxertos IAC 313 'Tropical' e IAC 766 'Campinas' não foi eficiente para a formação de mudas de videira.

Desenvolvimento, produtividade e sobrevivência de maracujazeiro-amarelo enxertado e cultivado em área com histórico de morte prematura de plantas

O objetivo deste trabalho foi avaliar o desenvolvimento vegetativo, a produtividade e a sobrevivência do maracujazeiro-amarelo (Passiflora edulis Sims) enxertado sobre três porta-enxertos, em área com histórico de morte prematura de plantas. O experimento foi conduzido no município de Adamantina-SP, no período de maio de 2006 a fevereiro de 2007, adotando-se o delineamento de blocos ao acaso, com três tratamentos e sete repetições. Os porta-enxertos avaliados foram Passiflora edulis, P. alata e P. gibertii, utilizando-se da enxertia convencional por garfagem tipo fenda cheia. Avaliaram-se o diâmetro do caule do porta-enxerto e do enxerto, o comprimento do entrenó e dos ramos secundários, o número de ramos terciários e o de frutos, a massa média, o diâmetro e o comprimento médio dos frutos, a produtividade e a sobrevivência de plantas. Os resultados demonstraram que o uso da enxertia no maracujazeiro é uma opção viável como meio de propagação vegetativa, assim como forma de controle de alguns patógenos habitantes do solo, um dos problemas que têm limitado a expansão da cultura. As plantas enxertadas sobre P. edulis apresentaram melhor desenvolvimento inicial, seguido de P. gibertii e de P. alata. A menor produtividade ocorrreu em plantas sobre P. alata. Mesmo com a presença de Fusarium solani e Rotylenchulus reniformis nos solos, 91% das plantas enxertadas sobre P. gibertii sobreviveram após 12 meses de plantio no campo, enquanto em P. alata e P. edulis, esses índices foram de 60% e 8,6%, respectivamente, mostrando assim a maior tolerância às doenças causadas por patógenos habitantes do solo por P. gibertii.

Alterações orais em crianças prematuras e de baixo peso ao nascer: a importância da relação entre pediatras e odontopediatras

OBJETIVO: Apresentar uma revisão de literatura sobre as alterações orais em crianças prematuras e de baixo peso ao nascer. FONTES DE DADOS: Foram selecionados artigos em inglês e português, desde 1976 até 2009, pesquisados no PubMed, Lilacs e na Bibliografia Brasileira de Odontologia (BBO), além de livros e consensos nacionais e internacionais. As palavras-chave utilizadas foram: prematuro, recém-nascido de baixo peso, cárie dentária, hipoplasia do esmalte dentário, manifestações bucais e dentição primária. Selecionaram-se artigos avaliando incidência, prevalência e etiologia das alterações orais, além de revisões de literatura e relatos de casos clínicos. SÍNTESE DOS DADOS: As alterações orais mais comuns em crianças prematuras e de baixo peso ao nascer são os defeitos no esmalte dentário (hipoplasias e hipocalcificações), a predisposição ao desenvolvimento de lesões de cárie dentária, as alterações na cronologia de erupção dentária e as alterações no palato com consequente aparecimento de maloclusão. Outras alterações também são relatadas, como diferenças nas dimensões das coroas dentárias e na espessura e porosidade do esmalte dentário. A interação entre pediatras e odontopediatras é fundamental no manejo dessas crianças. CONCLUSÕES: O conhecimento das alterações orais em crianças pré-termo e de baixo peso ao nascer por parte dos pediatras e odontopediatras favorece a atuação multidisciplinar com o objetivo de educar, prevenir e atenuar as possíveis mudanças físicas e dentárias nessas crianças.