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Article ; Online: Isolation of Microvesicles from Plasma Samples Avoiding Lipoprotein Contamination.

Merij, Laura B / Andrade, Fernanda B / Silva, Adriana R / Hottz, Eugenio D

Methods in molecular biology (Clifton, N.J.)

2021 Volume 2409, Page(s) 245–255

Abstract: Dengue is an infectious disease caused by Dengue Virus, mainly transmitted by Aedes aegypti mosquitoes. Severe dengue is a potentially fatal syndrome in consequence of overwhelmed inflammation, in which thrombocytopenia and increased vascular ... ...

Abstract	Dengue is an infectious disease caused by Dengue Virus, mainly transmitted by Aedes aegypti mosquitoes. Severe dengue is a potentially fatal syndrome in consequence of overwhelmed inflammation, in which thrombocytopenia and increased vascular permeability are frequently observed. Several experimental evidences point to the participation of both microvesicles (MVs) and circulating lipoproteins in inflammatory amplification in dengue pathogenesis. On this regard, many protocols for isolating plasma MVs have shown lipoproteins as the main contaminant. This is a limitation to studies aiming at the functional characterization of MVs, since both MVs and lipoproteins can modulate inflammatory responses. Here, we describe a biphasic density-based gradient ultracentrifugation as a tool for concomitant isolation of MVs and lipoproteins without cross-contamination. Flow cytometry for MVs quantification and western blot for detection of apoB100 may be used to confirm the isolation and purity of the MVs.
MeSH term(s)	Animals ; Cell-Derived Microparticles ; Dengue ; Flow Cytometry ; Lipoproteins
Chemical Substances	Lipoproteins
Language	English
Publishing date	2021-10-28
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-1879-0_17
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: The Weight of Obesity in Immunity from Influenza to COVID-19.

Andrade, Fernanda B / Gualberto, Ana / Rezende, Camila / Percegoni, Nathércia / Gameiro, Jacy / Hottz, Eugenio D

Frontiers in cellular and infection microbiology

2021 Volume 11, Page(s) 638852

Abstract: The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has emerged in December 2019 and rapidly outspread worldwide endangering human health. The coronavirus disease 2019 (COVID-19) manifests itself through a wide spectrum of symptoms that can ... ...

Abstract	The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has emerged in December 2019 and rapidly outspread worldwide endangering human health. The coronavirus disease 2019 (COVID-19) manifests itself through a wide spectrum of symptoms that can evolve to severe presentations as pneumonia and several non-respiratory complications. Increased susceptibility to COVID-19 hospitalization and mortality have been linked to associated comorbidities as diabetes, hypertension, cardiovascular diseases and, recently, to obesity. Similarly, individuals living with obesity are at greater risk to develop clinical complications and to have poor prognosis in severe influenza pneumonia. Immune and metabolic dysfunctions associated with the increased susceptibility to influenza infection are linked to obesity-associated low-grade inflammation, compromised immune and endocrine systems, and to high cardiovascular risk. These preexisting conditions may favor virological persistence, amplify immunopathological responses and worsen hemodynamic instability in severe COVID-19 as well. In this review we highlight the main factors and the current state of the art on obesity as risk factor for influenza and COVID-19 hospitalization, severe respiratory manifestations, extrapulmonary complications and even death. Finally, immunoregulatory mechanisms of severe influenza pneumonia in individuals with obesity are addressed as likely factors involved in COVID-19 pathophysiology.
MeSH term(s)	Adipokines ; Adipose Tissue ; Animals ; Body Weight ; COVID-19/immunology ; COVID-19/physiopathology ; Comorbidity ; Diabetes Mellitus ; Endotoxemia ; Heart Disease Risk Factors ; Hospitalization ; Humans ; Hyperglycemia ; Immunity ; Inflammation ; Influenza, Human/immunology ; Influenza, Human/physiopathology ; Metabolic Syndrome ; Obesity/complications ; Obesity/immunology ; Orthomyxoviridae Infections/immunology ; Risk Factors ; SARS-CoV-2
Chemical Substances	Adipokines
Language	English
Publishing date	2021-03-17
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2619676-1
ISSN	2235-2988 ; 2235-2988
ISSN (online)	2235-2988
ISSN	2235-2988
DOI	10.3389/fcimb.2021.638852
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Article ; Online: Role of aggregate-forming pilus (AFP) in adherence and colonization of both intestinal and urinary tracts.

Schüroff, Paulo A / Abe, Cecilia M / Silva, Jonatas W / de Paula Coelho, Cidéli / Andrade, Fernanda B / Hernandes, Rodrigo T / Dobrindt, Ulrich / Gomes, Tânia A T / Elias, Waldir P

Virulence

2022 Volume 13, Issue 1, Page(s) 1423–1433

Abstract: ... Hybrid- ... ...

Abstract	Hybrid-pathogenic
MeSH term(s)	Animals ; Bacterial Adhesion ; Escherichia coli/pathogenicity ; Escherichia coli Infections/microbiology ; Fimbriae, Bacterial ; HeLa Cells ; Humans ; Intestines/microbiology ; Mice ; Urinary Tract/microbiology ; Urinary Tract Infections/microbiology ; Uropathogenic Escherichia coli/pathogenicity
Language	English
Publishing date	2022-08-18
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2657572-3
ISSN	2150-5608 ; 2150-5594
ISSN (online)	2150-5608
ISSN	2150-5594
DOI	10.1080/21505594.2022.2112818
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Article ; Online: A sensitive and specific molecular tool for detection of both typical and atypical enteroaggregative Escherichia coli.

Andrade, Fernanda B / Gomes, Tânia A T / Elias, Waldir P

Journal of microbiological methods

2014 Volume 106, Page(s) 16–18

Abstract: A multiplex PCR has been standardized and evaluated for the identification of both typical and atypical enteroaggregative Escherichia coli (EAEC). The assay detecting aaiA, aaiG, aggR and aatA presented 94.8% of sensitivity, 94.3% of specificity and was ... ...

Abstract	A multiplex PCR has been standardized and evaluated for the identification of both typical and atypical enteroaggregative Escherichia coli (EAEC). The assay detecting aaiA, aaiG, aggR and aatA presented 94.8% of sensitivity, 94.3% of specificity and was able to efficiently detect both groups of EAEC among E. coli isolated from stool cultures.
MeSH term(s)	Adult ; Bacteriological Techniques/methods ; Child ; Child, Preschool ; Escherichia coli/genetics ; Escherichia coli/isolation & purification ; Escherichia coli Infections/diagnosis ; Escherichia coli Infections/microbiology ; Feces/microbiology ; Genes, Bacterial ; Humans ; Molecular Diagnostic Techniques/methods ; Multiplex Polymerase Chain Reaction/methods ; Sensitivity and Specificity
Language	English
Publishing date	2014-11
Publishing country	Netherlands
Document type	Evaluation Studies ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	604916-3
ISSN	1872-8359 ; 0167-7012
ISSN (online)	1872-8359
ISSN	0167-7012
DOI	10.1016/j.mimet.2014.07.030
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Article: A sensitive and specific molecular tool for detection of both typical and atypical enteroaggregative Escherichia coli

Andrade, Fernanda B / Tânia A.T. Gomes / Waldir P. Elias

Journal of microbiological methods. 2014 Nov., v. 106

2014

Abstract	A multiplex PCR has been standardized and evaluated for the identification of both typical and atypical enteroaggregative Escherichia coli (EAEC). The assay detecting aaiA, aaiG, aggR and aatA presented 94.8% of sensitivity, 94.3% of specificity and was able to efficiently detect both groups of EAEC among E. coli isolated from stool cultures.
Keywords	Escherichia coli ; feces ; microbial detection ; polymerase chain reaction
Language	English
Dates of publication	2014-11
Size	p. 16-18.
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	604916-3
ISSN	1872-8359 ; 0167-7012
ISSN (online)	1872-8359
ISSN	0167-7012
DOI	10.1016/j.mimet.2014.07.030
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Article ; Online: Distribution of serine protease autotransporters of Enterobacteriaceae in typical and atypical enteroaggregative Escherichia coli.

Andrade, Fernanda B / Abreu, Afonso G / Nunes, Kamila O / Gomes, Tânia A T / Piazza, Roxane M F / Elias, Waldir P

Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases

2017 Volume 50, Page(s) 83–86

Abstract: Enteroaggregative Escherichia coli (EAEC) is an agent of acute and persistent diarrhea worldwide, categorized in typical or atypical subgroups. Some EAEC virulence factors are members of the serine protease autotransporters of Enterobacteriaceae (SPATE). ...

Abstract	Enteroaggregative Escherichia coli (EAEC) is an agent of acute and persistent diarrhea worldwide, categorized in typical or atypical subgroups. Some EAEC virulence factors are members of the serine protease autotransporters of Enterobacteriaceae (SPATE). The presence of SPATE-encoding genes of different E. coli pathotypes was searched in a large collection of EAEC strains, and a possible association between SPATEs and E. coli phylogroups was investigated. Among 108 typical and 85 atypical EAEC, pic was the most prevalent gene, detected in 47.1% of the strains, followed by sat (24.3%), espI (21.2%), pet (19.2%), sepA (13.5%), sigA (4.1%), eatA (4.1%), vat (1.0%), espP and tsh, detected in one strain (0.5%) each; while epeA and espC were not detected. Phylogenetic analysis demonstrated that 39.9% of the strains belonged to group A, 23.3% to B1, 10.9% to B2, 7.8% to D, 8.8% to E and 1.5% to F. The majority of the SPATE genes were distributed in typical and atypical strains without association with any phylogroup. In addition, pic and pet were strongly associated with typical EAEC and sepA was detected in close association with atypical EAEC. Our data indicate that SPATEs may represent important virulence traits in both subgroups of EAEC.
MeSH term(s)	Adhesins, Escherichia coli/genetics ; Adhesins, Escherichia coli/metabolism ; Bacterial Toxins/genetics ; Bacterial Toxins/metabolism ; Carrier Proteins/genetics ; Carrier Proteins/metabolism ; Diarrhea/microbiology ; Diarrhea/pathology ; Enterotoxins/genetics ; Enterotoxins/metabolism ; Escherichia coli/classification ; Escherichia coli/enzymology ; Escherichia coli/genetics ; Escherichia coli/pathogenicity ; Escherichia coli Infections/microbiology ; Escherichia coli Infections/pathology ; Escherichia coli Proteins/genetics ; Escherichia coli Proteins/metabolism ; Gene Expression Regulation, Bacterial ; Genotype ; Humans ; Peptide Hydrolases/genetics ; Peptide Hydrolases/metabolism ; Phylogeny ; Serine Endopeptidases/genetics ; Serine Endopeptidases/metabolism ; Sigma Factor/genetics ; Sigma Factor/metabolism ; Virulence ; Virulence Factors/genetics ; Virulence Factors/metabolism
Chemical Substances	Adhesins, Escherichia coli ; Bacterial Toxins ; Carrier Proteins ; EatA protein, E coli ; Enterotoxins ; Escherichia coli Proteins ; Sigma Factor ; Virulence Factors ; secreted autotransporter toxin, E coli ; tsh protein, E coli ; vacuolating autotransporter toxin, E coli ; Peptide Hydrolases (EC 3.4.-) ; SepA protein, E coli (EC 3.4.-) ; EspP protein, E coli (EC 3.4.21.-) ; Pet protein, E coli (EC 3.4.21.-) ; Pic protein, E coli (EC 3.4.21.-) ; Serine Endopeptidases (EC 3.4.21.-)
Language	English
Publishing date	2017-06
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2037068-4
ISSN	1567-7257 ; 1567-1348
ISSN (online)	1567-7257
ISSN	1567-1348
DOI	10.1016/j.meegid.2017.02.018
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Article: Distribution of serine protease autotransporters of Enterobacteriaceae in typical and atypical enteroaggregative Escherichia coli

Andrade, Fernanda B / Afonso G. Abreu / Kamila O. Nunes / Roxane M.F. Piazza / Tânia A.T. Gomes / Waldir P. Elias

Infection, genetics, and evolution. 2017 June, v. 50

2017

Abstract	Enteroaggregative Escherichia coli (EAEC) is an agent of acute and persistent diarrhea worldwide, categorized in typical or atypical subgroups. Some EAEC virulence factors are members of the serine protease autotransporters of Enterobacteriaceae (SPATE). The presence of SPATE-encoding genes of different E. coli pathotypes was searched in a large collection of EAEC strains, and a possible association between SPATEs and E. coli phylogroups was investigated. Among 108 typical and 85 atypical EAEC, pic was the most prevalent gene, detected in 47.1% of the strains, followed by sat (24.3%), espI (21.2%), pet (19.2%), sepA (13.5%), sigA (4.1%), eatA (4.1%), vat (1.0%), espP and tsh, detected in one strain (0.5%) each; while epeA and espC were not detected. Phylogenetic analysis demonstrated that 39.9% of the strains belonged to group A, 23.3% to B1, 10.9% to B2, 7.8% to D, 8.8% to E and 1.5% to F. The majority of the SPATE genes were distributed in typical and atypical strains without association with any phylogroup. In addition, pic and pet were strongly associated with typical EAEC and sepA was detected in close association with atypical EAEC. Our data indicate that SPATEs may represent important virulence traits in both subgroups of EAEC.
Keywords	diarrhea ; Escherichia coli ; genes ; pathotypes ; pets ; phylogeny ; serine proteinases ; virulence
Language	English
Dates of publication	2017-06
Size	p. 83-86.
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	2037068-4
ISSN	1567-1348
ISSN	1567-1348
DOI	10.1016/j.meegid.2017.02.018
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Article ; Online: Structural Changes in Stx1 Engineering Monoclonal Antibody Improves Its Functionality as Diagnostic Tool for a Rapid Latex Agglutination Test.

Luz, Daniela / Shiga, Emerson A / Chen, Gang / Quintilio, Wagner / Andrade, Fernanda B / Maranhão, Andrea Q / Caetano, Bruna A / Mitsunari, Thaís / Silva, Míriam A / Rocha, Letícia B / Moro, Ana M / Sidhu, Sachdev S / Piazza, Roxane M F

Antibodies (Basel, Switzerland)

2018 Volume 7, Issue 1

Abstract: Stx1 toxin is one of the ... ...

Abstract	Stx1 toxin is one of the AB
Language	English
Publishing date	2018-02-01
Publishing country	Switzerland
Document type	Journal Article
ISSN	2073-4468
ISSN (online)	2073-4468
DOI	10.3390/antib7010009
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Article ; Online: Sensitive and specific detection of enteropathogenic and enterohemorrhagic Escherichia coli using recombinant anti-intimin antibody by immunofluorescence assay.

Caravelli, Andressa / Luz, Daniela E / Andrade, Fernanda B / Moraes, Claudia T P / Maranhão, Andrea Q / Piazza, Roxane M F

Diagnostic microbiology and infectious disease

2013 Volume 77, Issue 4, Page(s) 301–303

Abstract: The main and common virulence factor expressed by enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC) is intimin, a 94-kDa outer membrane protein, which is a product of the eae gene, and, thus, an excellent target for the ... ...

Abstract	The main and common virulence factor expressed by enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC) is intimin, a 94-kDa outer membrane protein, which is a product of the eae gene, and, thus, an excellent target for the detection of these pathogens. Among the methods for detection of virulence factor expression, immunoassays can be considered the first alternative to either animal use or in vitro culture cells assays, for which polyclonal and/or monoclonal antibodies are raised. In the present work, we evaluated the sensitivity and specificity of an intimin recombinant antibody (scFv-intimin) using immunofluorescence assay. The scFv-intimin detected typical EPEC, atypical EPEC, and EHEC isolates (100% sensitivity) with no detection of eae- isolates (100% specificity). Thus, immunofluorescence is an effective and rapid method, and scFv-intimin, an excellent tool for the diagnosis of diarrhea caused by EPEC and EHEC and also can be employed in case-control epidemiological surveys.
MeSH term(s)	Adhesins, Bacterial/immunology ; Enterohemorrhagic Escherichia coli/classification ; Enteropathogenic Escherichia coli/classification ; Escherichia coli Proteins/immunology ; Fluorescent Antibody Technique, Indirect/methods ; Humans ; Sensitivity and Specificity ; Single-Chain Antibodies/chemistry ; Single-Chain Antibodies/genetics ; Single-Chain Antibodies/immunology ; Virulence Factors/immunology
Chemical Substances	Adhesins, Bacterial ; Escherichia coli Proteins ; Single-Chain Antibodies ; Virulence Factors ; eaeA protein, E coli (147094-99-3)
Language	English
Publishing date	2013-12
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	604920-5
ISSN	1879-0070 ; 0732-8893
ISSN (online)	1879-0070
ISSN	0732-8893
DOI	10.1016/j.diagmicrobio.2013.08.021
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Article: Sensitive and specific detection of enteropathogenic and enterohemorrhagic Escherichia coli using recombinant anti-intimin antibody by immunofluorescence assay

Caravelli, Andressa / Luz, Daniela E / Andrade, Fernanda B / Moraes, Claudia T.P / Maranhão, Andrea Q / Piazza, Roxane M.F

Diagnostic microbiology & infectious disease. 2013 Dec., v. 77, no. 4

2013

Abstract	The main and common virulence factor expressed by enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC) is intimin, a 94-kDa outer membrane protein, which is a product of the eae gene, and, thus, an excellent target for the detection of these pathogens. Among the methods for detection of virulence factor expression, immunoassays can be considered the first alternative to either animal use or in vitro culture cells assays, for which polyclonal and/or monoclonal antibodies are raised. In the present work, we evaluated the sensitivity and specificity of an intimin recombinant antibody (scFv-intimin) using immunofluorescence assay. The scFv-intimin detected typical EPEC, atypical EPEC, and EHEC isolates (100% sensitivity) with no detection of eae− isolates (100% specificity). Thus, immunofluorescence is an effective and rapid method, and scFv-intimin, an excellent tool for the diagnosis of diarrhea caused by EPEC and EHEC and also can be employed in case–control epidemiological surveys.
Keywords	animals ; diarrhea ; enterohemorrhagic Escherichia coli ; enteropathogenic Escherichia coli ; epidemiological studies ; fluorescent antibody technique ; genes ; immunoassays ; in vitro culture ; intimin ; monoclonal antibodies ; pathogens ; rapid methods ; recombinant antibodies ; virulence
Language	English
Dates of publication	2013-12
Size	p. 301-303.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	604920-5
ISSN	0732-8893
ISSN	0732-8893
DOI	10.1016/j.diagmicrobio.2013.08.021
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