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3. Chromosomal Type II Toxin–Antitoxin Systems May Enhance Bacterial Fitness of a Hybrid Pathogenic Escherichia coli Strain Under Stress Conditions.

Author

Silva, Jessika C. A., Marques-Neto, Lazaro M., Carvalho, Eneas, Del Carpio, Alejandra M. G., Henrique, Camila, Leite, Luciana C. C., Mitsunari, Thais, Elias, Waldir P., Munhoz, Danielle D., and Piazza, Roxane M. F.

Subjects
ESCHERICHIA coli, GENOMICS, GENETIC transcription, BACTERIAL growth, CHROMOSOMES
Abstract

The functions of bacterial plasmid-encoded toxin–antitoxin (TA) systems are unambiguous in the sense of controlling cells that fail to inherit a plasmid copy. However, its role in chromosomal copies is contradictory, including stress-response-promoting fitness and antibiotic treatment survival. A hybrid pathogenic Escherichia coli strain may have the ability to colonize distinct host niches, facing contrasting stress environments. Herein, we determined the influence of multiple environmental stress factors on the bacterial growth dynamic and expression profile of previously described TA systems present in the chromosome of a hybrid atypical enteropathogenic and extraintestinal E. coli strain. Genomic analysis revealed 26 TA loci and the presence of five type II TA systems in the chromosome. Among the tested stress conditions, osmotic and acid stress significantly altered the growth dynamics of the hybrid strain, enhancing the necessary time to reach the stationary phase. Using qPCR analyses, 80% of the studied TA systems were differentially expressed in at least one of the tested conditions, either in the log or in the stationary phase. These data indicate that type II TA systems may contribute to the physiology of pathogenic hybrid strains, enabling their adaptation to different milieus. [ABSTRACT FROM AUTHOR]

Published
2024
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4. Genomic Dissection of an Enteroaggregative Escherichia coli Strain Isolated from Bacteremia Reveals Insights into Its Hybrid Pathogenic Potential.

Author

Del Carpio, Alejandra M. G., Freire, Claudia A., Andrade, Fernanda B., Piazza, Roxane M. F., Silva, Rosa M., Carvalho, Eneas, and Elias, Waldir P.

Subjects
ESCHERICHIA coli, WHOLE genome sequencing, INTESTINAL infections, BACTEREMIA, GENETIC markers
Abstract

Escherichia coli is a frequent pathogen isolated from bloodstream infections. This study aimed to characterize the genetic features of EC092, an E. coli strain isolated from bacteremia that harbors enteroaggregative E. coli (EAEC) genetic markers, indicating its hybrid pathogenic potential. Whole-genome sequencing showed that EC092 belongs to phylogroup B1, ST278, and serotype O165:H4. Genes encoding virulence factors such as fimbriae, toxins, iron-uptake systems, autotransporter proteins (Pet, Pic, Sat, and SepA), and secretion systems were detected, as well as EAEC virulence genes (aggR, aatA, aaiC, and aap). EC092 was found to be closely related to the other EAEC prototype strains and highly similar in terms of virulence to three EAEC strains isolated from diarrhea. The genomic neighborhood of pet, pic, sat, sepA, and the EAEC virulence genes of EC092 and its three genetically related fecal EAEC strains showed an identical genomic organization and nucleotide sequences. Also, EC092 produced and secreted Pet, Pic, Sat, and SepA in the culture supernatant and resisted the bactericidal activity of normal human serum. Our results demonstrate that the strain EC092, isolated from bacteremia, is a hybrid pathogenic extraintestinal E. coli (ExPEC)/EAEC with virulence features that could mediate both extraintestinal and intestinal infections. [ABSTRACT FROM AUTHOR]

Published
2024
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9. SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time Validation

Author

Coelho, Fabiana Fioravante, primary, da Silva, Miriam Aparecida, additional, Lopes, Thiciany Blener, additional, Polatto, Juliana Moutinho, additional, de Castro, Natália Salazar, additional, Andrade, Luis Adan Flores, additional, Lourenço, Karine Lima, additional, Sato, Hugo Itaru, additional, de Carvalho, Alex Fiorini, additional, Coelho, Helena Perez, additional, Bagno, Flávia Fonseca, additional, Luz, Daniela, additional, Viala, Vincent Louis, additional, Cattony, Pedro Queiroz, additional, Melo, Bruna de Sousa, additional, Moro, Ana Maria, additional, Quintilio, Wagner, additional, Barbosa, Ana Paula, additional, Bomfim, Camila Gasque, additional, Soares, Camila Pereira, additional, Guzzo, Cristiane Rodrigues, additional, Fonseca, Flavio Guimarães, additional, Durigon, Edison Luiz, additional, Gazzinelli, Ricardo Tostes, additional, Ribeiro Teixeira, Santuza M., additional, Piazza, Roxane Maria Fontes, additional, and Fernandes, Ana Paula, additional

Published
2023
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10. A new class of antimicrobial molecules derived from kefir, effective against Pseudomonas aeruginosa and methicillin resistant Staphylococcus aureus (MRSA) strains

Author

Marques, Vaniky Duarte, Franzolin, Marcia Regina, Sanabani, Sabri Saeed, Vigerelli, Hugo, Piazza, Roxane Maria Fontes, Pimenta, Daniel Carvalho, Venâncio, Tiago, Neves, Irys Viana, de Sousa Silva, Herbert Guimarães, dos Santos Courrol, Daniella, Mendonça-Previato, Lucia, Previato, José Osvaldo, Jorge, Soraia Attie Calil, and de Oliveira Domingos, Marta

Published
2020
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13. Diarrheagenic Escherichia coli

Author

Gomes, Tânia A.T., Elias, Waldir P., Scaletsky, Isabel C.A., Guth, Beatriz E.C., Rodrigues, Juliana F., Piazza, Roxane M.F., Ferreira, Luís C.S., and Martinez, Marina B.

Published
2016
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16. Identification of Zika Virus NS1-Derived Peptides with Potential Applications in Serological Tests

Author

Prudencio, Carlos Roberto, primary, Gomes da Costa, Vivaldo, additional, Rocha, Leticia Barboza, additional, Costa, Hernan Hermes Monteiro, additional, Orts, Diego José Belato, additional, da Silva Santos, Felipe Rocha, additional, Rahal, Paula, additional, Lino, Nikolas Alexander Borsato, additional, da Conceição, Pâmela Jóyce Previdelli, additional, Bittar, Cintia, additional, Machado, Rafael Rahal Guaragna, additional, Durigon, Edison Luiz, additional, Araujo, João Pessoa, additional, Polatto, Juliana Moutinho, additional, da Silva, Miriam Aparecida, additional, de Oliveira, Joyce Araújo, additional, Mitsunari, Thais, additional, Pereira, Lennon Ramos, additional, Andreata-Santos, Robert, additional, de Souza Ferreira, Luís Carlos, additional, Luz, Daniela, additional, and Piazza, Roxane Maria Fontes, additional

Published
2023
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17. O55 Polysaccharides Are Good Antigen Targets for the Formulation of Vaccines against O55 STEC and Capsulated aEPEC Strains

Author

Silva, Herbert Guimarães de Sousa, primary, Franzolin, Marcia Regina, additional, Anjos, Geovana Ferreira dos, additional, Barbosa, Angela Silva, additional, Santos, Luis Fernando dos, additional, Miranda, Kaique Ferrari, additional, Marques, Ronaldo Maciel, additional, Souza, Matilde Costa Lima de, additional, Piazza, Roxane Maria Fontes, additional, and Domingos, Marta de Oliveira, additional

Published
2022
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21. Recombinant PilS: Cloning, Expression and Biochemical Characterization of a Pil-Fimbriae Subunit

Author

Munhoz, Danielle D., primary, Silva, Jessika C. A., additional, Freitas, Natalia C., additional, Iwai, Leo K., additional, Aires, Karina A., additional, Ozaki, Christiane Y., additional, Souza, Cristiane S., additional, Rocha, Letícia B., additional, Silva, Miriam A., additional, Henrique, Izabella M., additional, Elias, Waldir P., additional, Carvalho, Eneas, additional, Morganti, Ligia, additional, Chura-Chambi, Rosa M., additional, and Piazza, Roxane M. F., additional

Published
2022
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22. Escherichia coli Strains Responsible for Cystitis in Female Pediatric Patients with Normal and Abnormal Urinary Tracts Have Different Virulence Profiles

Author

Domingos, Marta, primary, da Silva Junior, Silvio, additional, Milanello, Wagner, additional, Nakano, Shirley, additional, Franzolin, Marcia, additional, dos Santos, Luis, additional, Nunes, Kamila, additional, Marques, Vaniky, additional, Elias, Waldir, additional, Silva, Herbert, additional, Caetano, Bruna, additional, and Piazza, Roxane, additional

Published
2022
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24. In Silico Prediction and Design of Uropathogenic Escherichia coli Alpha-Hemolysin Generate a Soluble and Hemolytic Recombinant Toxin

Author

Caetano, Bruna De Lucca, primary, Domingos, Marta de Oliveira, additional, da Silva, Miriam Aparecida, additional, da Silva, Jessika Cristina Alves, additional, Polatto, Juliana Moutinho, additional, Montoni, Fabio, additional, Iwai, Leo Kei, additional, Pimenta, Daniel Carvalho, additional, Vigerelli, Hugo, additional, Vieira, Paulo Cesar Gomes, additional, Ruiz, Rita de Cassia, additional, Patané, José Salvatore, additional, and Piazza, Roxane Maria Fontes, additional

Published
2022
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25. The Deleterious Effects of Shiga Toxin Type 2 Are Neutralized In Vitro by FabF8:Stx2 Recombinant Monoclonal Antibody

Author

Luz, Daniela, primary, Gómez, Fernando D., additional, Ferreira, Raíssa L., additional, Melo, Bruna S., additional, Guth, Beatriz E. C., additional, Quintilio, Wagner, additional, Moro, Ana Maria, additional, Presta, Agostina, additional, Sacerdoti, Flavia, additional, Ibarra, Cristina, additional, Chen, Gang, additional, Sidhu, Sachdev S., additional, Amaral, María Marta, additional, and Piazza, Roxane M. F., additional

Published
2021
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26. Role of the YehD fimbriae in the virulence‐associated properties of enteroaggregativeEscherichia coli

Author

Soria‐Bustos, Jorge, primary, Saitz, Waleska, additional, Medrano, Abraham, additional, Lara‐Ochoa, Cristina, additional, Bennis, Zineb, additional, Monteiro‐Neto, Valerio, additional, dos Santos, Camilla Itapary, additional, Rodrigues, Josias, additional, Hernandes, Rodrigo T., additional, Yáñez, Jorge A., additional, Torres, Javier, additional, Navarro‐García, Fernando, additional, Martínez‐Laguna, Ygnacio, additional, Fontes Piazza, Roxane M., additional, Munhoz, Danielle Dias, additional, Cedillo, María L., additional, Ares, Miguel A., additional, De la Cruz, Miguel A., additional, Nataro, James P., additional, and Girón, Jorge A., additional

Published
2021
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27. Encontro de Panstrongylus megistus em ecótopo artificial: domiciliação ou mera visitação?

Author

Nascimento Carlos, Marassá Ana M., Curado Izilda, and Piazza Roxane M.F.

Subjects
P. megistus, Domiciliação, Trypanosoma cruzi, Arctic medicine. Tropical medicine, RC955-962
Abstract

No combate à doença de Chagas no Brasil, foi utilizado como primeira medida o controle químico, erradicando o Triatoma infestans, o mais importante vetor, tal combate vem favorecendo o aparecimento de vetores secundários, como Triatoma sordida and Panstrongylus megistus, espécies que podem eventualmente ser encontradas no domicílio, como os triatomíneos provenientes de Bernardino de Campos e Sete Barras por nós examinados que foram encontrados no domicílio e positivos para o T. cruzi, sugerindo que, apesar da doença de Chagas estar controlada no Estado de São Paulo, existe a necessidade de aprimorar os conhecimentos sobre o comportamento destes vetores para que mudanças nas medidas de controle sejam introduzidas.

Published
1997

34. Macrophage inflammatory response mediated by intimin and bundle-forming pilus from enteropathogenic <em>Escherichia coli</em>

Author

Borges, Monamaris, primary, Piazza, Roxane, primary, Caetano*, Bruna, additional, Mourão*, Daniela, additional, Abreu, Patricia, additional, Monaris, Denize, additional, Galhardo, Cynthia, additional, Menezes, Marcio, additional, Vasconcellos, Halyka, additional, Luz, Daniela, additional, Lima, Flavia, additional, and Elias, Waldir, additional

Published
2020
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35. Paracoccidioides HSP90 Can Be Found in the Cell Surface and Is a Target for Antibodies with Therapeutic Potential

Author

Moura, Ágata, primary, Oliveira, Diane, additional, Paredes, Verenice, additional, Rocha, Letícia, additional, Oliveira, Fabiana, additional, Lessa, Gustavo, additional, Riasco-Palacios, Juan, additional, Casadevall, Arturo, additional, Albuquerque, Patrícia, additional, Felipe, Maria, additional, Piazza, Roxane, additional, and Nicola, André, additional

Published
2020
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37. Paracoccidioides HSP90 can be found in the cell surface and is a target for antibodies with therapeutic potential

Author

Moura, Ágata Nogueira D’Áurea, primary, de Oliveira, Diane Sthefany Lima, additional, Paredes, Verenice, additional, Rocha, Letícia Barboza, additional, Casadevall, Arturo, additional, Albuquerque, Patrícia, additional, Felipe, Maria Sueli Soares, additional, Piazza, Roxane Maria Fontes, additional, and Nicola, André Moraes, additional

Published
2020
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38. Surface Protein Dispersin of Enteroaggregative Escherichia coli Binds Plasminogen That Is Converted Into Active Plasmin

Author

Moraes, Claudia T. P., primary, Longo, Jonathan, additional, Silva, Ludmila B., additional, Pimenta, Daniel C., additional, Carvalho, Eneas, additional, Morone, Mariana S. L. C., additional, da Rós, Nancy, additional, Serrano, Solange M. T., additional, Santos, Ana Carolina M., additional, Piazza, Roxane M. F., additional, Barbosa, Angela S., additional, and Elias, Waldir P., additional

Published
2020
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41. Role of the YehD fimbriae in the virulence‐associated properties of enteroaggregative Escherichia coli.

Author

Soria‐Bustos, Jorge, Saitz, Waleska, Medrano, Abraham, Lara‐Ochoa, Cristina, Bennis, Zineb, Monteiro‐Neto, Valerio, dos Santos, Camilla Itapary, Rodrigues, Josias, Hernandes, Rodrigo T., Yáñez, Jorge A., Torres, Javier, Navarro‐García, Fernando, Martínez‐Laguna, Ygnacio, Fontes Piazza, Roxane M., Munhoz, Danielle Dias, Cedillo, María L., Ares, Miguel A., De la Cruz, Miguel A., Nataro, James P., and Girón, Jorge A.

Subjects
ESCHERICHIA coli, CELL adhesion, BACTERIAL cell surfaces, HELA cells, SPINACH, INFLAMMATION
Abstract

Summary: The interaction of enteroaggregative Escherichia coli (EAEC) strains with the colonic gut mucosa is characterized by the ability of the bacteria to form robust biofilms, to bind mucin, and induce a local inflammatory response. These events are mediated by a repertoire of five different aggregative adherence fimbriae variants (AAF/I‐V) typically encoded on virulence plasmids. In this study, we report the production in EAEC strains of a new YehD fimbriae (YDF), which is encoded by the chromosomal gene cluster yehABCD, also present in most E. coli strains. Immuno‐labelling of EAEC strain 042 with anti‐AAF/II and anti‐YDF antibodies demonstrated the presence of both AAF/II and YDF on the bacterial surface. We investigated the role of YDF in cell adherence, biofilm formation, colonization of spinach leaves, and induction of pro‐inflammatory cytokines release. To this aim, we constructed yehD deletion mutants in different EAEC backgrounds (strains 17‐2, 042, 55989, C1010, 278‐1, J7) each harbouring one of the five AAFs. The effect of the YDF mutation was strain dependent and AAF independent as the lack of YDF had a different impact on the phenotypes manifested by the different EAECs tested. Expression of the yehABCD operon in a E. coli K12 ORN172 showed that YDF is important for biofilm formation but not for adherence to HeLa cells. Lastly, screening of pro‐inflammatory cytokines in supernatants of Caco‐2 cells infected with EAEC strains 042 and J7 and their isogenic ΔyehD mutants showed that these mutants were significantly defective in release of IL‐8 and TNF‐α. This study contributes to the understanding of the complex and diverse mechanisms of adherence of EAEC strains and identifies a new potential target for preventive measures of gastrointestinal illness caused by EAEC and other E. coli pathogroups. [ABSTRACT FROM AUTHOR]

Published
2022
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44. Characterization of the universal stress protein F from atypical enteropathogenic Escherichia coli and its prevalence in Enterobacteriaceae

Author

de Souza, Cristiane S., Torres, Alfredo G., Caravelli, Andressa, Silva, Anderson, Polatto, Juliana M., and Piazza, Roxane M. F.

Subjects
Escherichia coli Proteins, cloning, α/β protein, Shigella sonnei, hypothetical proteins, Articles, Article, Citrobacter freundii, stress, Enteropathogenic Escherichia coli, expression, UspF protein, Heat-Shock Proteins, atypical EPEC
Abstract

Atypical enteropathogenic Escherichia coli (aEPEC) are heterogeneous strains in terms of serotypes, adherence patterns and the presence of novel virulence factors. This heterogeneity is intriguing, promoting studies trying to characterize these novel proteins and to better comprehend this pathotype group. In a previous study analyzing low‐molecular mass proteomes of four representative aEPEC strains of three different adhesion phenotypes, we classified proteins according to their annotated function, with most of them being involved in metabolism and transport; while some of them were classified as hypothetical proteins. The majority of the hypothetical proteins were homologue products of genes identified in the genome of enterohemorrhagic E. coli. One of the hypothetical proteins was annotated as Z2335, with orthologue in EPEC, and by bioinformatics analysis, this protein was revealed to be the universal stress protein F (UspF). Thus, herein we successfully obtained a recombinant UspF protein from aEPEC, which is a α/β, ATP‐binding protein involved in stress response, with comparable protein production among the four studied strains, but showing noteworthy differences when cultivated in different stress conditions, also present in other enterobacterial species, such as Shigella sonnei and Citrobacter freundii. Furthermore, our results confirm that the Usp protein superfamily encompasses a conserved group of proteins involved in stress resistance in aEPEC and other Enterobacteriaceae.

Published
2016

46. Macrophage Inflammatory Response Mediated by Intimin and Bundle-Forming Pilus from Enteropathogenic Escherichia coli

Author

Caetano, Bruna A., primary, Mourão, Daniela B., additional, Abreu, Patrícia A. E., additional, Monaris, Denize, additional, Vasconcellos, Halyka L., additional, Luz, Daniela, additional, Galhardo, Cynthia Soares, additional, Menezes, Marcio A., additional, Lima, Flávia A., additional, Elias, Waldir P., additional, Borges, Monamaris Marques, additional, and Piazza, Roxane Maria Fontes, additional

Published
2020
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48. The Type III Secretion System (T3SS)-Translocon of Atypical Enteropathogenic Escherichia coli (aEPEC) Can Mediate Adherence

Author

Santos, Fernanda F., primary, Yamamoto, Denise, additional, Abe, Cecilia M., additional, Bryant, Jack A., additional, Hernandes, Rodrigo T., additional, Kitamura, Felipe C., additional, Castro, Felipe S., additional, Valiatti, Tiago B., additional, Piazza, Roxane M. F., additional, Elias, Waldir P., additional, Henderson, Ian R., additional, and Gomes, Tânia A. T., additional

Published
2019
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49. Cloning approach and functional analysis of anti-intimin single-chain variable fragment (scFv)

Author

Elias Waldir P, Abreu Patrícia AE, Pereira Milton CA, Ruiz Renato M, Ozaki Christiane Y, Aires Karina A, Menezes Márcio A, Ramos Oscar HP, and Piazza Roxane MF

Subjects
Medicine, Biology (General), QH301-705.5, Science (General), Q1-390
Abstract

Abstract Background Intimin is an important virulence factor involved in the pathogenesis of enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic Escherichia coli (EHEC). Both pathogens are still important causes of diarrhea in children and adults in many developing and industrialized countries. Considering the fact that antibodies are important tools in the detection of various pathogens, an anti-intimin IgG2b monoclonal antibody was previously raised in immunized mice with the conserved sequence of the intimin molecule (int388-667). In immunoblotting assays, this monoclonal antibody showed excellent specificity. Despite good performance, the monoclonal antibody failed to detect some EPEC and EHEC isolates harboring variant amino acids within the 338-667 regions of intimin molecules. Consequently, motivated by its use for diagnosis purposes, in this study we aimed to the cloning and expression of the single-chain variable fragment from this monoclonal antibody (scFv). Findings Anti-intimin hybridoma mRNA was extracted and reversely transcripted to cDNA, and the light and heavy chains of the variable fragment of the antibody were amplified using commercial primers. The amplified chains were cloned into pGEM-T Easy vector. Specific primers were designed and used in an amplification and chain linkage strategy, obtaining the scFv, which in turn was cloned into pAE vector. E. coli BL21(DE3)pLys strain was transformed with pAE scFv-intimin plasmid and subjected to induction of protein expression. Anti-intimin scFv, expressed as inclusion bodies (insoluble fraction), was denatured, purified and submitted to refolding. The protein yield was 1 mg protein per 100 mL of bacterial culture. To test the functionality of the scFv, ELISA and immunofluorescence assays were performed, showing that 275 ng of scFv reacted with 2 mg of purified intimin, resulting in an absorbance of 0.75 at 492 nm. The immunofluorescence assay showed a strong reactivity with EPEC E2348/69. Conclusion This study demonstrated that the recombinant anti-intimin antibody obtained is able to recognize the conserved region of intimin (Int388-667) in purified form and the EPEC isolate.

Published
2011
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