Cloning and Identification of a Complete cDNA Coding for a Bactericidal and Antitumoral Acidic Phospholipase A(2) from Bothrops jararacussu Venom

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

New mariner elements in Anastrepha species (Diptera: Tephritidae)

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

Molecular Characterization and Physical Mapping of Two Classes of 5S rDNA in the Genomes of Gymnotus sylvius and G. inaequilabiatus (Gymnotiformes, Gymnotidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Molecular organization of 5S rDNA in sharks of the genus Rhizoprionodon: insights into the evolutionary dynamics of 5S rDNA in vertebrate genomes

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

Distinct localization of T cell Agrin during antigen presentation - evidence for the expression of Agrin receptor(s) in antigen-presenting cells

Agrin is over-expressed by activated and autoimmune T cells, and synergizes with the T cell receptor (TCR) to augment cell activation. In the present study, we show that Agrin accumulates to distinct areas of the plasma membrane and that cell activation causes its redistribution. During antigen presentation, Agrin primarily accumulates to the periphery of the mature immunological synapse, mostly in lamellipodia-like protrusions that wrap around the antigen-presenting cell and, conversely, anti-Agrin sera induced a significant redistribution of TCR at the plasma membrane. We also provide evidence for the expression of Agrin receptors in peripheral blood monocytes, dendritic cells and a fraction of B cells. Interestingly, interferon-a treatment, which induces the expression of Agrin in T cells, also augmented Agrin binding to monocytes. Stimulation of monocytes with recombinant Agrin induced the clustering of surface receptors, including major histocompatibility complex class II, activation of intracellular signalling cascades, as well as enhanced dsRNA-induced expression of pro-inflammatory cytokines interleukin-6 and tumour necrosis factor-a. Collectively, these results confirm the location of Agrin at the immunological synapse between T cells and antigen-presenting cells and justify further characterization of its receptors in the immune system.

Losing identity: structural diversity of transposable elements belonging to different classes in the genome of Anopheles gambiae

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Amino acid sequence of a myotoxic Lys49-phospholipase A(2) homologue from the venom of Cerrophidion (Bothrops) godmani

The complete amino acid sequence of myotoxin II (godMT-II), a myotoxic phospholipase A( 2 )(PLA(2)) homologue from the venom of the Central American crotaline snake Cerrophidion (Bothrops) godmani, was determined by direct protein sequencing methods. GodMT-II is a class II PLA, showing a Lys instead of Asp at position 49. An additional substitution in the calcium binding loop region (Asn instead of Tyr at position 28) suggests the lack of enzymatic activity observed in this toxin is due to loss of its ability to bind the co-factor Ca2+, since the residues involved in forming the catalytic network of PLA(2)s (His-48, Tyr-52 and Asp-99) an conserved in godMT-II. This myotoxin shows highest sequence homology with other Lys-49 PLA(2)s from Bothrops, Agkistrodon and Trimeresurus species, suggesting that they constitute a conserved family of proteins, yet in contrast presents lower homology with Bothrops asper myotoxin III, a catalytically-active PLA(2). The C-terminal region of godMT-II, which is rich in cationic and hydrophobic residues, shares high sequence homology to the corresponding region in the myotoxin II from B. asper, which has been proposed to play an important role in the Ca2+-independent membrane damaging activity. (C) 1998 Elsevier B.V. B.V. All rights reserved.

Activation of Ca2+-independent membrane-damaging activity in Lys49-phospholipase A(2) promoted by amphiphilic molecules

Association of class-II phospholipase A(2) (PLA(2)) with aggregated phospholipid substrate results in elevated levels of the Ca2+-dependent hydrolytic activity. The Asp49 residue participates in coordination of the Ca2+ ion cofactor, however, in Lys49-PLA(2) homologues (Lys49-PLA(2)S), substitution of the Asp49 by Lys results in loss of Ca2+ binding and lack of detectable phospholipid hydrolysis. Nevertheless, Lys49-PLA2S cause Ca2+-independent damage of liposome membranes. Bothropstoxin-I is a homodimeric Lys49-PLA(2) from the venom of Bothrops jararacussu, and in fluorescent marker release and dynamic light scattering experiments with DPPC liposomes we demonstrate activation of the Ca2+-independent membrane damaging activity by similar to4 molecules of sodium dodecyl sulphate (SDS) per protein monomer. Activation is accomparlied by significant changes in the intrinsic tryptophan fluorescence emission (ITFE) and near UV circular dichroism (UVCD) spectra of the protein. Subsequent binding of 7-10 SDS molecules results in further alterations in the ITFE and far UVCD spectra. Reduction in the rate of N-bromosuccinimide modification of Trp77 at the dimer interface suggests that initial binding of SDS to this region accompanies the activation of the membrane damaging activity. 1-anilinonaphthalene-8-sulphonic acid binding studies indicate that subsequent SDS binding to the active site is concomitant with the second structural transition. These results provide insights in the structural basis of amphiphile/protein coupling in class-II PLA(2)s. (C) 2004 Published by Elsevier B.V.

Structural insights into the catalytic mechanism of sphingomyelinases D and evolutionary relationship to glycerophosphodiester phosphodiesterases

Spider venom sphingomyelinases D catalyze the hydrolysis of sphingomyelin via an Mg2+ ion-dependent acid-base catalytic mechanism which involves two histidines. In the crystal structure of the sulfate free enzyme determined at 1.85 angstrom resolution, the metal ion is tetrahedrally coordinated instead of the trigonal-bipyramidal coordination observed in the sulfate bound form. The observed hyperpolarized state of His47 requires a revision of the previously suggested catalytic mechanism. Molecular modeling indicates that the fundamental structural features important for catalysis are fully conserved in both classes of SMases D and that the Class II SMases D contain an additional intra-chain disulphide bridge (Cys53-Cys201). Structural analysis suggests that the highly homologous enzyme from Loxosceles bonetti is unable to hydrolyze sphingomyelin due to the 95G1y -> Asn and 134Pro -> Glu mutations that modify the local charge and hydrophobicity of the interfacial face. Structural and sequence comparisons confirm the evolutionary relationship between sphingomyelinases D and the glicerophosphodiester phosphoesterases which utilize a similar catalytic mechanism. (c) 2006 Elsevier B.V. All rights reserved.

Os efeitos dos aparelhos funcionais sobre a dimensão transversal da maxila e mandíbula

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influence of the banded Herbst appliance on dental changes in mixed dentition

OBJECTIVE: This prospective clinical study was conducted with the purpose of evaluating the influence of the banded Herbst appliance on dental changes during the early treatment of Class II malocclusion. METHOD: The sample consisted of 15 prepubertal subjects (12 boys and 3 girls, initial age: 9 years and 6 months) who were treated with the Herbst appliance. Treatment effects were compared with those of a Class II Division 1 group of 15 subjects (8 boys and 7 girls, mean initial age 9 years and 1 month), not treated orthodontically. Statistical analysis was performed using Student t-test with 5% significance level. RESULTS: The results showed that treatment with the banded Herbst appliance in the mixed dentition stage tended to upright maxillary incisors (mean: 4.14°). The maxillary molars were distalized and intruded significantly (mean 2.65 mm and 1.24 mm, respectively), the lower incisors slightly protruded anteriorly (mean 1.64 mm) and the molars showed no significant changes in the horizontal and vertical directions. Furthermore, significant improvements were noted in overbite (1.26 mm), overjet (4.8 mm) and molar relationship (12.08 mm). CONCLUSIONS: Changes in the upper dental arch were found to be greater than changes in the lower arch. Furthermore, mandibular anchorage loss was reduced due to the anchorage system used in the study.

Cephalometric evaluation of skeletal alterations induced by Herbst appliance during mixed dentition

OBJECTIVE: The present prospective clinical study was designed in order to evaluate horizontal and vertical skeletal alterations induced by the use of Herbst appliance in individuals with Class II, division 1 malocclusion during mixed dentition stage. METHODS: The sampling consisted of 15 pre-pubertal individuals (12 boys and 3 girls; initial age 9 years and 6 months), who were treated with Herbst appliance for a period of 7 months. The effects of the treatment were compared to a group of 15 individuals with Class II, division 1 malocclusion (8 boys and 7 girls, initial age averaged 9 years and 1 month), orthodontically untreated, who were followed up for a period of 12 months. Statistical analysis was performed with Student's t-test with significance level at 5%. RESULTS: It was showed that the treatment with Herbst appliance in mixed dentition stage has restricted maxilla growth. Mandibular and palatal planes have not undergone significant alteration; however, anterior and posterior facial heights have increased significantly. Facial convexity and maxillomandibular relationship were altered positively. Mandible has positioned significantly forward and its effective length increased 2.5 times more than the increase observed in control group. CONCLUSION: It was possible to conclude that Herbst appliance was able to provide satisfactory results in individuals during mixed dentition stage.

Influence of intentional ankylosis of deciduous canines to reinforce the anchorage for maxillary protraction

INTRODUÇÃO: nesse estudo cefalométrico retrospectivo, analisou-se a influência da anquilose intencional de caninos decíduos em pacientes com má oclusão de Classe III e mordida cruzada anterior, nos estágios de dentição decídua e mista precoce, tratados com expansão ortopédica da maxila, seguida de tração reversa. MÉTODOS: foram utilizadas telerradiografias em norma lateral de 40 pacientes, divididos em 2 grupos pareados por idade e sexo. O Grupo Anquilose foi constituído de 20 pacientes (10 meninos e 10 meninas) tratados com anquilose induzida e que apresentavam as idades médias inicial e final, respectivamente, de 7a 4m e 8a 3m, e o tempo médio de tração reversa de 11 meses. O Grupo Controle, composto de 20 pacientes (10 meninos e 10 meninas) tratados sem anquilose induzida e que apresentavam as idades médias inicial de 7a 8m e final de 8a 7m, e tempo médio de tração reversa de 11 meses. Foram empregadas as análises de Variância a dois critérios e de Covariância para comparar as variáveis cefalométricas inicial e final e as alterações de tratamento entre os grupos. RESULTADOS: segundo os resultados, as variáveis que evidenciaram as mudanças de tratamento significativas entre os grupos confirmaram que o procedimento de anquilose intencional potencializou a resposta sagital das bases apicais (Pg-NPerp) e aumentou os ângulos de convexidade facial (NAP e ANB). CONCLUSÃO: o protocolo envolvendo a anquilose intencional de caninos decíduos potencializou a resposta sagital das bases apicais.

Occurrence of skeletal malocclusions in Brazilian patients with dentofacial deformities

Este estudo avaliou a ocorrência de más oclusões esqueléticas apresentadas pelos pacientes do Centro de Pesquisa e Tratamento das Deformidades Bucofaciais (CEDEFACE), na cidade de Araraquara, SP, Brasil. Foram avaliados prontuários de 381 pacientes com deformidades dentoesqueléticas, que fizeram tratamento combinado ortodôntico-cirúrgico no período entre 2000 e 2006. Após a seleção da amostra (método de conveniência), baseado nos dados da documentação pré e pós-cirúrgica, o número de pacientes foi reduzido para 171. Para classificação do levantamento, considerou-se a discrepância ântero-posterior (Classe I, II e III), raça, idade, gênero, ausência ou presença de assimetria, excesso vertical maxilar e biprotrusão maxilar, além de determinar em qual base óssea o procedimento cirúrgico foi realizado. As documentações dos pacientes foram analisadas por um examinador previamente calibrado pelo processo de repetição até que o método fosse considerado adequado (correlação intraclasse >0,94). A idade média dos pacientes foi de 23,59 anos (DP 6,93), a maioria do gênero feminino (102 pacientes) e leucoderma (160 pacientes). A má oclusão mais prevalente foi a Classe III (81 pacientes). A assimetria, o excesso maxilar vertical e biprotrusão maxilar estavam presentes em 54, 33, e 7 pacientes, respectivamente. Na maioria dos casos, as cirurgias para correção de deformidades dentoesqueléticas foram combinadas, envolvendo os dois maxilares. Com base nos resultados, conclui-se que a Classe III foi a deformidade esquelética mais prevalente e a Classe I a menos prevalente. em geral, a prevalência de deformidades esqueléticas foi maior entre as mulheres e a maioria dos pacientes apresentou uma combinação de problemas maxilares e mandibulares, o que interfere diretamente na decisão sobre o plano de tratamento mais adequado. Houve uma maior incidência de assimetria na Classe III esquelética; o excesso vertical ocorreu de forma semelhante na Classe II e III e a biprotrusão teve baixa incidência entre as más oclusões avaliadas.

Efeitos dentoesqueléticos produzidos pelo aparelho de Herbst na dentadura mista

OBJETIVO: o objetivo desta pesquisa clínica prospectiva foi avaliar as alterações cefalométricas dentárias e esqueléticas produzidas pelo aparelho de Herbst em jovens com má oclusão de Classe II, 1ª divisão durante a dentadura mista. METODOLOGIA: trinta jovens (15 do gênero masculino e 15 do feminino) com idade média inicial de 9 anos e 10 meses foram tratados com o aparelho de Herbst por um período de 12 meses. Para a comparação dos grupos utilizou-se uma amostra controle de 30 jovens (15 do gênero masculino e 15 do feminino) Classe II, 1ª divisão, com idade média inicial de 9 anos e 8 meses, que foram mantidos sem tratamento durante 12 meses. Para cada jovem foram utilizadas duas telerradiografias em norma lateral, obtidas ao início e no final do período de acompanhamento. Utilizou-se um método convencional de avaliação cefalométrica e o método proposto por Pancherz. RESULTADOS E CONCLUSÕES: os resultados deste estudo demonstraram que os efeitos do aparelho de Herbst produzidos na dentadura mista foram primariamente de natureza dentoalveolar. Os incisivos inferiores foram inclinados para vestibular e os superiores foram retruídos; também houve uma extrusão significante dos molares inferiores, enquanto os superiores sofreram restrição de desenvolvimento no sentido vertical. Não houve diferença significante de restrição do crescimento anterior da maxila entre os dois grupos. No sentido vertical da face, a altura facial ântero-inferior se comportou de forma similar, não demonstrando alteração significante entre os grupos. O tratamento com o aparelho de Herbst produziu um aumento modesto, porém, significante no comprimento da mandíbula comparado ao grupo controle. Este aumento, entretanto, foi de menor magnitude que aquele observado em pacientes adolescentes utilizando o mesmo protocolo de tratamento. A correção do overjet (Herbst) ocorreu devido a 22% de alterações esqueléticas e 78% de alterações dentárias. A correção da relação molar ocorreu devido a 27% de alterações esqueléticas e 73% de alterações dentárias.