Wanderley Dias da Silveira

Universidade Estadual de Campinas (UNICAMP)

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Co-Authors

Marcelo Brocchi

Universidade Estadual de Campinas (UNICAMP)

Eliana Guedes Stehling

Universidade de São Paulo

Gerson Nakazato

Universidade Estadual de Londrina

Antônio Fernando Pestana de Castro

Universidade de São Paulo

Thaís Cabrera Galvão Rojas

Universidade Estadual de Campinas (UNICAMP)

Renato Pariz Maluta

Universidade Estadual de Campinas (UNICAMP)

Tatiana Amabile de Campos

Universidade de Brasília

Fernanda de Pace

Boston Children's Hospital

Jacqueline Boldrin de Paiva

Universidade Estadual de Campinas (UNICAMP)

Marcelo Lancellotti

Universidade Estadual de Campinas (UNICAMP)

Cooperative Institutions

Universidade Estadual de Campinas (UNICAMP)

Universidade de São Paulo

Hospital de Clínicas da Unicamp

Universidade Estadual Paulista (Unesp)

Universidade Estadual de Londrina

Instituto Adolfo Lutz

Universidade Federal de São Paulo

Universidade de Ribeirão Preto

Fundação Oswaldo Cruz

Wellcome Sanger Institute

Author Statistics

Papers

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Cloning and Purification of IpaC Antigen from Shigella flexneri: Proposal of a New Methodology

Protein and Peptide Letters (2012)

Cristiane Mobilon Marcelo A. S. Toledo Fernanda Laroza Paganelli Clelton A. Santos Fernanda de Pace

Shigella flexneri is a Gram-negative bacillus that is responsible for a severe form of dysentery called Shigellosis, which mainly affects children and the elderly in both underdeveloped and developed countries. Pathogenic S. flexneri strains possess a large virulence plasmid that codes for effector proteins that are required for the entry and spread of the bacteria into colonocytes. Among these proteins is the translocator IpaC, which plays an important role in the invasion process; IpaC is implicated in pore formation in the host cell membrane and induces cytoskeletal rearrangements in macrophages and epithelial cells, thereby promoting bacterial entry. The ability of IpaC to insert onto the plasma membrane is due to a large nonpolar region of the protein structure. This characteristic also renders difficulties in recovery and purification when the protein is expressed in E. coli. Several works have considered different methodologies for the improved production and purification of IpaC. Herein, we propose an alternative method that is based on changes in the induction temperature and extraction buffer to facilitate the accumulation of high yields of soluble proteins for their further processing and ultimate use in biotechnological approaches.

Shigellosis

Thaumatin

Cloning (programming)

Protein purification

10.2174/092986613804725316

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Citations (3)

In silico phylogenetic and virulence gene profile analyses of avian pathogenic Escherichia coli genome sequences

Pesquisa Veterinária Brasileira (2014)

Thaís Cabrera Galvão Rojas Renato Pariz Maluta Luciano Vieira Koenigkan Wanderley Dias da Silveira

Avian pathogenic Escherichia coli (APEC) infections are responsible for significant losses in the poultry industry worldwide. A zoonotic risk has been attributed to APEC strains because they present similarities to extraintestinal pathogenic E. coli (ExPEC) associated with illness in humans, mainly urinary tract infections and neonatal meningitis. Here, we present in silico analyses with pathogenic E. coli genome sequences, including recently available APEC genomes. The phylogenetic tree, based on multi-locus sequence typing (MLST) of seven housekeeping genes, revealed high diversity in the allelic composition. Nevertheless, despite this diversity, the phylogenetic tree was able to cluster the different pathotypes together. An in silico virulence gene profile was also determined for each of these strains, through the presence or absence of 83 well-known virulence genes/traits described in pathogenic E. coli strains. The MLST phylogeny and the virulence gene profiles demonstrated a certain genetic similarity between Brazilian APEC strains, APEC isolated in the United States, UPEC (uropathogenic E. coli) and diarrheagenic strains isolated from humans. This correlation corroborates and reinforces the zoonotic potential hypothesis proposed to APEC.

Multilocus sequence typing

Pathogenic Escherichia coli

Housekeeping gene

10.1590/s0100-736x2014000200006

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Citations (3)

Involvement of the Helicobacter pylori plasticity region and cag pathogenicity island genes in the development of gastroduodenal diseases

European Journal of Clinical Microbiology & Infectious Diseases (2008)

Alline R. Pacheco José Luiz Proença-Módena Ana Isabela L. Sales-Moioli Y. Fukuhara Wanderley Dias da Silveira

CagA

Pathogenicity island

Peptic

Medical microbiology

Spirillaceae

10.1007/s10096-008-0549-8

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Citations (32)

Molecular typing and biological characteristics of Pseudomonas aeruginosa isolated from cystic fibrosis patients in Brazil

The Brazilian Journal of Infectious Diseases (2010)

Eliana Guedes Stehling Domingos da Silva Leite Wanderley Dias da Silveira

The present study had as objective to evaluate the genotypic diversity and biological characteristics, such as hemolysin, protease, elastase of 56 clinical strains of Pseudomonas aeruginosa isolated from 13 cystic fibrosis (CF) patients attending at the School Hospital of Campinas State University (UNICAMP), Brazil. Genotypic diversity has been determined by Ribotyping (RT) and the pattern of the enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) of each strain. The production of elastase was significantly different only among mucoid and nonmucoid isolates. Joint results obtained by (RT) and ERIC-PCR methods were able to discriminate all strains isolated from both the same and different patients. Additionally, we observed four strain clusters with low diversity. The most infective strains were located in just two clusters. These results suggest that either there is a strong selection towards a specific genotype or that specific isolates could be responsible for the initial and subsequent colonization processes. More studies are necessary to know if these conclusions can be generalized for the general CF population.

Ribotyping

Intergenic region

Strain (injury)

10.1016/s1413-8670(10)70094-5

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Citations (32)

Development of a bacterial cloning vector for expression of scorpion toxins for biotechnological studies

Protein Expression and Purification (2007)

Eliana Guedes Stehling Wanderley Dias da Silveira Tatiana Amabile de Campos Marcelo Brocchi Thelma A. Pertinhez

Cloning (programming)

Cloning vector

10.1016/j.pep.2007.08.001

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Citations (7)

Levels of Expression and Immunogenicity of Attenuated Salmonella enterica Serovar Typhimurium Strains Expressing Escherichia coli Mutant Heat-Labile Enterotoxin

Infection and Immunity (1998)

M. Giuseppina Covone Marcelo Brocchi Emanuela Palla Wanderley Dias da Silveira Rino Rappuoli

ABSTRACT The effects of heterologous gene dosage as well as Salmonella typhimurium strain variability on immune response toward both the heterologous antigen, the nontoxic mutant of the Escherichia coli heat-labile enterotoxin LTK63, and the carrier Salmonella strain have been analyzed. Effects of a single integration into the host DNA and different-copy-number episomal vectors were compared in S. typhimurium Δcya Δcrp Δasd strains of two different serotypes, UK-1 and SR-11. Expression of the enterotoxin in the different Salmonella isolates in vitro was found to vary considerably and, for the episomal vectors, to correlate with the plasmid copy number. LTK63-specific serum immunoglobulin G (IgG) and mucosal immunoglobulin A (IgA) antibodies were highest in mice immunized with the high-level-expression strain. High anti-LTK63 IgG and IgA titers were found to correspond to higher anti- Salmonella immunity, suggesting that LTK63 exerts an adjuvant effect on response to the carrier. Statistically significant differences in anti-LTK63 immune response were observed between groups of mice immunized with the attenuated Δ cya Δcrp UK-1 and SR-11 derivatives producing the antigen at the same rate. These data indicate that the same attenuation in S. typhimurium strains of different genetic backgrounds can influence significantly the immune response toward the heterologous antigen. Moreover, delivery of the LTK63 enterotoxin to the immune system by attenuated S. typhimurium strains is effective only when synthesis of the antigen is very high during the initial phase of invasion, while persistence of the S. typhimurium strain in deep tissues has only marginal influence.

Heat-labile enterotoxin

Heterologous

Salmonella enterica

10.1128/iai.66.1.224-231.1998

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Citations (55)

Cloning and characterization of the gene encoding the OmpU outer membrane protein of Vibrio cholerae

Infection and Immunity (1996)

Vanessa Sperandio Camella Bailey Jorge A. Girón Victor J. DiRita Wanderley Dias da Silveira

The OmpU outer membrane protein is a member of the ToxR regulon of Vibrio cholerae and has recently been shown to be a potential adherence factor for this species. Using PCR and degenerate oligonucleotide primers based on internal peptide sequences of purified OmpU, we have cloned and sequenced the gene encoding OmpU. The ompU gene is predicted to encode a 36,646-molecular-weight protein which is present in both cholera toxin-positive and -negative V. cholerae O1 and O139 strains.

Regulon

Cholera toxin

Cloning (programming)

10.1128/iai.64.12.5406-5409.1996

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Citations (55)

Molecular typing and biological characteristics of Pseudomonas aeruginosa isolated from cystic fibrosis patients in Brazil

The Brazilian Journal of Infectious Diseases (2010)

Eliana Guedes Stehling Domingos da Silva Leite Wanderley Dias da Silveira

Ribotyping

Intergenic region

Strain (injury)

10.1590/s1413-86702010000500007

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Citations (10)

The expression of plasmid mediated afimbrial adhesin genes in an avian septicemic Escherichia coli strain

Journal of Veterinary Science (2008)

Eliana Guedes Stehling Tatiana Amabile de Campos Marcelo Brocchi Vasco Azevedo Wanderley Dias da Silveira

An Escherichia coli strain (SEPT13) isolated from the liver of a hen presenting clinical signs of septicaemia had a LD50 of 4.0 × 10 5 CFU/ml in one-day-old chickens, expressed Ia, Ib, E1, E3, K and B colicins and aerobactin.The strain was ampicillin and streptomycin resistant, and found to have fimA, csgA and tsh DNA related sequences; it could adhere to and invade HEp-2 and tracheal epithelial cells, expressed fimbriae (observed by electron microscopy), and had five plasmids of 2.7, 4.7, 43, 56, and 88 MDa.Transposon mutagenesis of strain SEPT13, with transposon TnphoA, resulted in a mutant strain named ST16 that had a LD50 of 1.2 × 10 12 CFU/ml.All other biological characteristics of strain ST16 were the same as those detected for strain SEPT13 except for the migration of an 88 MDa plasmid to the 93 MDa position indicating the insertion of the transposon into the 88 MDa plasmid.The 93 MDa plasmid of strain ST16 was transferred, by electroporation assay, to non-pathogenic receptor strains (E. coli strains K12 MS101 and HB101), resulting in transformant strains A and B, respectively.These strains exhibited adhesion properties to in vitro cultivated HEp-2 cells but did not have the capacity for invasion.The adherence occurred despite the absence of fimbriae; this finding suggests that the 88 MDa plasmid has afimbrial adhesin genes.

Strain (injury)

Transposon mutagenesis

Colicin

Aerobactin

10.4142/jvs.2008.9.1.75

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Citations (3)

Genome-Wide Survey of Genes Under Positive Selection in Avian PathogenicEscherichia coliStrains

Foodborne Pathogens and Disease (2017)

Thaís Cabrera Galvão Rojas Francisco Pereira Lobo Jorge Augusto Hongo Renato Vicentini Renu Verma

The ability to obtain bacterial genomes from the same host has allowed for comparative studies that help in the understanding of the molecular evolution of specific pathotypes. Avian pathogenic Escherichia coli (APEC) is a group of extraintestinal strains responsible for causing colibacillosis in birds. APEC is also suggested to possess a role as a zoonotic agent. Despite its importance, APEC pathogenesis still has several cryptic pathogenic processes that need to be better understood. In this work, a genome-wide survey of eight APEC strains for genes with evidence of recombination revealed that ∼14% of the homologous groups evaluated present signs of recombination. Enrichment analyses revealed that nine Gene Ontology (GO) terms were significantly more represented in recombinant genes. Among these GO terms, several were noted to be ATP-related categories. The search for positive selection in these APEC genomes revealed 32 groups of homologous genes with evidence of positive selection. Among these groups, we found several related to cell metabolism, as well as several uncharacterized genes, beyond the well-known virulence factors ompC, lamB, waaW, waaL, and fliC. A GO term enrichment test showed a prevalence of terms related to bacterial cell contact with the external environment (e.g., viral entry into host cell, detection of virus, pore complex, bacterial-type flagellum filament C, and porin activity). Finally, the genes with evidence of positive selection were retrieved from genomes of non-APEC strains and tested as were done for APEC strains. The result revealed that none of the groups of genes presented evidence of positive selection, confirming that the analysis was effective in inferring positive selection for APEC and not for E. coli in general, which means that the study of the genes with evidence of positive selection identified in this study can contribute for the better understanding of APEC pathogenesis processes.

Pathogenic Escherichia coli

10.1089/fpd.2016.2219

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Citations (8)