Hyaluronidase from the venom of the social wasp Polybia paulista (Hymenoptera, Vespidae): Cloning, structural modeling, purification, and immunological analysis

In this study, we describe the cDNA cloning, sequencing, and 3-D structure of the allergen hyaluronidase from Polybia paulista venom (Pp-Hyal). Using a proteomic approach, the native form of Pp-Hyal was purified to homogeneity and used to produce a Pp-specific polyclonal antibody. The results revealed that Pp-Hyal can be classified as a glycosyl hydrolase and that the full-length Pp-Hyal cDNA (1315 bp; GI: 302201582) is similar (80-90%) to hyaluronidase from the venoms of endemic Northern wasp species. The isolated mature protein is comprised of 338 amino acids, with a theoretical pI of 8.77 and a molecular mass of 39,648.8 Da versus a pI of 8.13 and 43,277.0 Da indicated by MS. The Pp-Hyal 3D-structural model revealed a central core (α/β)7 barrel, two sulfide bonds (Cys 19-308 and Cys 185-197), and three putative glycosylation sites (Asn79, Asn187, and Asn325), two of which are also found in the rVes v 2 protein. Based on the model, residues Ser299, Asp107, and Glu109 interact with the substrate and potential epitopes (five conformational and seven linear) located at surface-exposed regions of the structure. Purified native Pp-Hyal showed high similarity (97%) with hyaluronidase from Polistes annularis venom (Q9U6V9). Immunoblotting analysis confirmed the specificity of the Pp-Hyal-specific antibody as it recognized the Pp-Hyal protein in both the purified fraction and P. paulista crude venom. No reaction was observed with the venoms of Apis mellifera, Solenopsis invicta, Agelaia pallipes pallipes, and Polistes lanio lanio, with the exception of immune cross-reactivity with venoms of the genus Polybia (sericea and ignobilis). Our results demonstrate cross-reactivity only between wasp venoms from the genus Polybia. The absence of cross-reactivity between the venoms of wasps and bees observed here is important because it allows identification of the insect responsible for sensitization, or at least of the phylogenetically closest insect, in order to facilitate effective immunotherapy in allergic patients. © 2013 Elsevier Ltd.

Age and Morphological Changes in the Epiponini Wasp Polybia paulista Von Ihering (Hymenoptera: Vespidae)

The division of labor is a central theme in the study of social insects. In bees and wasps, this activity is regulated by age polyethism. Important physiological and morphological changes have been widely studied in the polyethism of honeybee workers. In contrast, this is a relatively unexplored subject in social vespids. Our goal was to determine if there are detectable morphological changes in the body of the Epiponini wasp Polybia paulista Von Ihering or in certain glands in relation to age polyethism. We observed changes in the body weight, the salivary gland, and the mandibular gland that were associated with age, and our results suggest that social relationships and task performance are important to these changes. This contrasts with observations in Polistes and is different from the Apis mellifera Linnaeus age polyethism model. © 2013 Sociedade Entomológica do Brasil.

Deputados estaduais e democracia: Valores políticos das elites parlamentares paulista e paranaense

After 20 years of reestablished democracy in Brazil, what do the state legislators think about the regime? The goal of the article is to show the views of state legislators of the Legislative Assemblies of the States of São Paulo and Paraná on democracy. For the views we use a structured and self-administered questionnaire. The results show that for the state legislators: i) democracy is stable, ii) it is superior to any other kind of regime, iii) it's support comes from some public policy and the use of voting and iv) parties and elections are important for democracy.

Frequência de ocorrência de cavidade anoftálmica na região centro-oeste paulista e características dos portadores

Purpose: To observe the frequency of the occurrence of the anophthalmic socket in the Middle West region of the state of São Paulo and to describe the demographic profile of the carriers in a population-based data. Methods: A cross-sectional study involve a random sampling carried out in twelve cities of the Middle West region of the state of São Paulo, for which the reference center is the city of Botucatu was done. The participators were chosen by assortment which considered the houses of these people. It was established a sampling with 11,453 people. All the exams were realized using a Mobile Ophthalmologic Unit. The research protocol included the demographic data and the complete ophthalmologic exam (anamnesis, ocular and systemic antecedents, familiar antecedents, visual acuity with and without correction, tonometry, biomicroscopy, fundoscopy, and refraction exam). All the data were transferred to an Excel spreadsheet and submitted to a descriptive analysis and were presented by the frequency of the occurrence. Results: Ophthalmic socket frequency in the Middle West region of the state of São Paulo was 0.96%. We found 11 cases of anophthalmic socket, with involvement of 0.7 % in females and 1.3 %, male. The most common causes of anophthalmic socket were glaucoma (blind painful eye), microphthalmia, trauma, and endophthalmitis. The majority of the people were 40 years old or more, and we found just one person younger than 19 years old. Conclusion: The anophthalmic socket occurred in 0.96% of the habitants of the Middle West region of São Paulo State, occurring mainly in male and with large variety of the affected ages.

Cytotoxic, genotoxic/antigenotoxic and mutagenic/antimutagenic effects of the venom of the wasp Polybia paulista

Hymenoptera venoms are constituted by a complex mixture of chemically or pharmacologically bioactive agents, such as phospholipases, hyaluronidases and mastoparans. Venoms can also contain substances that are able to inhibit and/or diminish the genotoxic or mutagenic action of other compounds that are capable of promoting damages in the genetic material. Thus, the present study aimed to assess the effect of the venom of Polybia paulista, a neotropical wasp, by assays with HepG2 cells maintained in culture. The cytotoxic potential of the wasp venom, assessed by the methyl thiazolyl tetrazolium assay (MTT assay), was tested for the concentrations of 10μg/mL, 5μg/mL and 1μg/mL. As these concentrations were not cytotoxic, they were used to evaluate the genotoxic (comet assay) and mutagenic potential (micronucleus test) of the venom. In this study, it was verified that these concentrations induced damages in the DNA of the exposed cells, and it was necessary to test lower concentrations until it was found those that were not considered genotoxic and mutagenic. The concentrations of 1ng/mL, 100pg/mL and 10pg/mL, which did not induce genotoxicity and mutagenicity, were used in four different treatments (post-treatment, pre-treatment, simultaneous treatment with and without incubation), in order to evaluate if these concentrations were able to inhibit or decrease the genotoxic and mutagenic action of methyl methanesulfonate (MMS). None of the concentrations was able to inhibit and/or decrease the MMS activity. The genotoxic and mutagenic activity of the venom of P. paulista could be caused by the action of phospholipase, mastoparan and hyaluronidase, which are able to disrupt the cell membrane and thereby interact with the genetic material of the cells or even facilitate the entrance of other compounds of the venom that can act on the DNA. Another possible explanation for the genotoxicity and mutagenicity of the venom can be the presence of substances able to trigger inflammatory process and, consequently, generate oxygen reactive species that can interact with the DNA of the exposed cells. © 2013 Elsevier Ltd.

Structure-function relationships of the peptide Paulistine: A novel toxin from the venom of the social wasp Polybia paulista

Background: The peptide Paulistine was isolated from the venom of wasp Polybia paulista. This peptide exists under a natural equilibrium between the forms: oxidised - with an intra-molecular disulphide bridge; and reduced - in which the thiol groups of the cysteine residues do not form the disulphide bridge. The biological activities of both forms of the peptide are unknown up to now. Methods: Both forms of Paulistine were synthesised and the thiol groups of the reduced form were protected with the acetamidemethyl group [Acm-Paulistine] to prevent re-oxidation. The structure/activity relationships of the two forms were investigated, taking into account the importance of the disulphide bridge. Results: Paulistine has a more compact structure, while Acm-Paulistine has a more expanded conformation. Bioassays reported that Paulistine caused hyperalgesia by interacting with the receptors of lipid mediators involved in the cyclooxygenase type II pathway, while Acm-Paullistine also caused hyperalgesia, but mediated by receptors involved in the participation of prostanoids in the cyclooxygenase type II pathway. Conclusion: The acetamidemethylation of the thiol groups of cysteine residues caused small structural changes, which in turn may have affected some physicochemical properties of the Paulistine. Thus, the dissociation of the hyperalgesy from the edematogenic effect when the actions of Paulistine and Acm-Paulistine are compared to each other may be resulting from the influence of the introduction of Acm-group in the structure of Paulistine. General significance: The peptides Paulistine and Acm-Paulistine may be used as interesting tools to investigate the mechanisms of pain and inflammation in future studies. © 2013 Elsevier B.V.

Leitura do fotojornalismo na escola: uma experiência com alunos da escola básica

Pós-graduação em Educação - FFC

A gramaticalização de então no português paulista: um estudo pancrônico

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

As vogais médias pretônicas dos verbos na fala culta do interior paulista

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

Variação lexical e sintática na produção escrita formal do português em Moçambique

Pós-graduação em Estudos Linguísticos - IBILCE

A haplologia na variedade paulista

Pós-graduação em Estudos Linguísticos - IBILCE

As vogais pretônicas na fala culta do Noroeste Paulista

Pós-graduação em Estudos Linguísticos - IBILCE

Fototriangulação radical analítica: como uma ferramenta de ensino de fototriangulação e de ajustamento de observações

Pós-graduação em Ciências Cartográficas - FCT

Formação continuada do professor de arte: dialogia com a experiência

Pós-graduação em Artes - IA

Adote o artista não deixe ele virar professor: reflexões em torno do híbrido professor performer

Pós-graduação em Artes - IA