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Article ; Online: Transmission Electron Microscopy Studies of Dengue Viruses.

Caldas, Gabriela Cardoso / Barth, Ortrud Monika / Barreto-Vieira, Debora Ferreira

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

2021 Volume 2409, Page(s) 11–30

Abstract: Despite the advancement of molecular biology techniques, morphological studies using transmission electron microscopy (TEM) are still being of great importance to elucidate some aspects of viral structures, morphogenesis, and pathogenesis. In relation to ...

Abstract	Despite the advancement of molecular biology techniques, morphological studies using transmission electron microscopy (TEM) are still being of great importance to elucidate some aspects of viral structures, morphogenesis, and pathogenesis. In relation to dengue viruses (DENV), several studies report the use of TEM to obtain a clearer definition of viral morphology, the events involved in its morphogenesis, aspects of pathogenesis, and cell tropism. In this chapter, the main technical protocols and their respective reagents for studies of DENV infection by TEM are described, both in cell culture and in biological tissue samples.
MeSH term(s)	Dengue Virus ; Microscopy, Electron, Transmission
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_2
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Monkeypox (Mpox) virus isolation and ultrastructural characterisation from a Brazilian human sample case.

Miranda, Milene Dias / Caldas, Gabriela Cardoso / Ferreira, Vivian Neuza / Barth, Ortrud Monika / da Silva, Aline de Paula Dias / Silva, Mayara Secco Torres / Grinsztejn, Beatriz / Veloso, Valdiléa Gonçalves / Souza, Thiago Moreno / da Silva, Edson Elias / Barreto-Vieira, Debora Ferreira

Memorias do Instituto Oswaldo Cruz

2023 Volume 118, Page(s) e230090

Abstract: Background: According to the last 2023 Monkeypox (Mpox) Outbreak Global Map from the Centres for Disease Control and Prevention (CDC), more than 100 countries with no Mpox infection report cases. Brazil stands out in this group and is the second country ...

Abstract	Background: According to the last 2023 Monkeypox (Mpox) Outbreak Global Map from the Centres for Disease Control and Prevention (CDC), more than 100 countries with no Mpox infection report cases. Brazil stands out in this group and is the second country with the highest number of cases in the last outbreak. Objective: To contribute to knowledge of the virus infection effects in a cellular model, which is important for diagnosis infections not yet included in a provider´s differential diagnosis and for developing viral inhibition strategies. Methods: We describe a virus isolation protocol for a human clinical sample from a patient from Brazil, the viral growth in a cell model through plaque forming units (PFU) assay, reverse transcriptase polymerase chain reaction (RT-PCR) and transmission electron microscopy (TEM). Findings: We follow the viral isolation in Vero cell culture from a Mpox positive clinically diagnosed sample and show the infection effects on cellular structures using a TEM. Main conclusions: Understanding the impact of viral growth on cellular structures and its replication kinetics may offer better strategies for the development of new drugs with antiviral properties.
MeSH term(s)	Humans ; Brazil ; Mpox (monkeypox) ; Biological Assay ; Diagnosis, Differential ; Disease Outbreaks
Language	English
Publishing date	2023-08-28
Publishing country	Brazil
Document type	Journal Article
ZDB-ID	953293-6
ISSN	1678-8060 ; 0074-0276
ISSN (online)	1678-8060
ISSN	0074-0276
DOI	10.1590/0074-02760230090
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Article ; Online: Dengue, Yellow Fever, Zika and Chikungunya epidemic arboviruses in Brazil: ultrastructural aspects.

Barreto-Vieira, Debora Ferreira / Couto-Lima, Dinair / Jácome, Fernanda Cunha / Caldas, Gabriela Cardoso / Barth, Ortrud Monika

Memorias do Instituto Oswaldo Cruz

2021 Volume 115, Page(s) e200278

Abstract: Background: The impact of arbovirus cocirculation in Brazil is unknown. Dengue virus (DENV) reinfection may result in more intense viraemia or immunopathology, leading to more severe disease. The Zika virus (ZIKV) epidemic in the Americas provided ... ...

Abstract	Background: The impact of arbovirus cocirculation in Brazil is unknown. Dengue virus (DENV) reinfection may result in more intense viraemia or immunopathology, leading to more severe disease. The Zika virus (ZIKV) epidemic in the Americas provided pathogenicity evidence that had not been previously observed in flavivirus infections. In contrast to other flaviviruses, electron microscopy studies have shown that ZIKV may replicate in viroplasm-like structures. Flaviviruses produce an ensemble of structurally different virions, collectively contributing to tissue tropism and virus dissemination. Objectives and methods: In this work, the Aedes albopictus mosquito cell lineage (C6/36 cells) and kidney epithelial cells from African green monkeys (Vero cells) were infected with samples of the main circulating arboviruses in Brazil [DENV-1, DENV-2, DENV-3, DENV-4, ZIKV, Yellow Fever virus (YFV) and Chikungunya virus (CHIKV)], and ultrastructural studies by transmission electron microscopy were performed. Findings: We observed that ZIKV, the DENV serotypes, YFV and CHIKV particles are spherical. ZIKV, DENV-1, -2, -3 and -4 presented diameters of 40-50 nm, and CHIKV presented approximate diameters of 50-60 nm. Viroplasm-like structures was observed in ZIKV replication cycle. Main conclusions: The morphogenesis of these arboviruses is similar to what has been presented in previous studies. However, we understand that further studies are needed to investigate the relationship between viroplasm-like structures and ZIKV replication dynamics.
MeSH term(s)	Animals ; Arboviruses ; Brazil/epidemiology ; Chikungunya Fever/epidemiology ; Chlorocebus aethiops ; Dengue/epidemiology ; Epidemics ; Vero Cells ; Yellow Fever ; Zika Virus ; Zika Virus Infection/epidemiology
Language	English
Publishing date	2021-02-03
Publishing country	Brazil
Document type	Journal Article
ZDB-ID	953293-6
ISSN	1678-8060 ; 0074-0276
ISSN (online)	1678-8060
ISSN	0074-0276
DOI	10.1590/0074-02760200278
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Article ; Online: Primary infection of BALB/c mice with a dengue virus type 4 strain leads to kidney injury.

Rasinhas, Arthur da Costa / Jácome, Fernanda Cunha / Caldas, Gabriela Cardoso / de Almeida, Ana Luisa Teixeira / de Souza, Daniel Dias Coutinho / Dos Santos, João Paulo Rodrigues / Dias, Helver Gonçalves / Araujo, Eduarda Lima / Mohana-Borges, Ronaldo / Barth, Ortrud Monika / Dos Santos, Flavia Barreto / Barreto-Vieira, Debora Ferreira

Memorias do Instituto Oswaldo Cruz

2023 Volume 118, Page(s) e220255

Abstract: Background: Dengue is a disease caused by dengue virus (DENV-1 through -4). Among the four serotypes, DENV-4 remains the least studied. Acute kidney injury is a potential complication of dengue generally associated with severe dengue infection.: ... ...

Abstract	Background: Dengue is a disease caused by dengue virus (DENV-1 through -4). Among the four serotypes, DENV-4 remains the least studied. Acute kidney injury is a potential complication of dengue generally associated with severe dengue infection. Objectives: The goal of this study was to investigate the alterations caused by experimental dengue infection in the kidney of adult BALB/c mice. Methods: In this study, BALB/c mice were infected through the intravenous route with a DENV-4 strain, isolated from a human patient. The kidneys of the mice were procured and subject to histopathological and ultrastructural analysis. Findings: The presence of the viral antigen was confirmed through immunohistochemistry. Analysis of tissue sections revealed the presence of inflammatory cell infiltrate throughout the parenchyma. Glomerular enlargement was a common find. Necrosis of tubular cells and haemorrhage were also observed. Analysis of the kidney on a transmission electron microscope allowed a closer look into the necrotic tubular cells, which presented nuclei with condensed chromatin, and loss of cytoplasm. Main conclusions: Even though the kidney is probably not a primary target of dengue infection in mice, the inoculation of the virus in the blood appears to damage the renal tissue through local inflammation.
MeSH term(s)	Adult ; Humans ; Animals ; Mice ; Dengue Virus ; Kidney ; Antigens, Viral ; Mice, Inbred BALB C ; Severe Dengue
Chemical Substances	Antigens, Viral
Language	English
Publishing date	2023-05-08
Publishing country	Brazil
Document type	Journal Article
ZDB-ID	953293-6
ISSN	1678-8060 ; 0074-0276
ISSN (online)	1678-8060
ISSN	0074-0276
DOI	10.1590/0074-02760220255
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Article ; Online: Inhibition of the SREBP pathway prevents SARS-CoV-2 replication and inflammasome activation.

Soares, Vinicius Cardoso / Dias, Suelen Silva Gomes / Santos, Julia Cunha / Azevedo-Quintanilha, Isaclaudia G / Moreira, Isabela Batista Gonçalves / Sacramento, Carolina Q / Fintelman-Rodrigues, Natalia / Temerozo, Jairo R / da Silva, Marcos Alexandre Nunes / Barreto-Vieira, Debora Ferreira / Souza, Thiago Ml / Bozza, Patricia T

Life science alliance

2023 Volume 6, Issue 11

Abstract: SARS-CoV-2 induces major cellular lipid rearrangements, exploiting the host's metabolic pathways to replicate. Sterol regulatory element binding proteins (SREBPs) are a family of transcription factors that control lipid metabolism. SREBP1 is associated ... ...

Abstract	SARS-CoV-2 induces major cellular lipid rearrangements, exploiting the host's metabolic pathways to replicate. Sterol regulatory element binding proteins (SREBPs) are a family of transcription factors that control lipid metabolism. SREBP1 is associated with the regulation of fatty acids, whereas SREBP2 controls cholesterol metabolism, and both isoforms are associated with lipid droplet (LD) biogenesis. Here, we evaluated the effect of SREBP in a SARS-CoV-2-infected lung epithelial cell line (Calu-3). We showed that SARS-CoV-2 infection induced the activation of SREBP1 and SREBP2 and LD accumulation. Genetic knockdown of both SREBPs and pharmacological inhibition with the dual SREBP activation inhibitor fatostatin promote the inhibition of SARS-CoV-2 replication, cell death, and LD formation in Calu-3 cells. In addition, we demonstrated that SARS-CoV-2 induced inflammasome-dependent cell death by pyroptosis and release of IL-1β and IL-18, with activation of caspase-1, cleavage of gasdermin D1, was also reduced by SREBP inhibition. Collectively, our findings help to elucidate that SREBPs are crucial host factors required for viral replication and pathogenesis. These results indicate that SREBP is a host target for the development of antiviral strategies.
MeSH term(s)	Humans ; Inflammasomes ; SARS-CoV-2 ; Sterol Regulatory Element Binding Protein 1 ; COVID-19 ; Lipid Metabolism
Chemical Substances	Inflammasomes ; Sterol Regulatory Element Binding Protein 1
Language	English
Publishing date	2023-09-05
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2575-1077
ISSN (online)	2575-1077
DOI	10.26508/lsa.202302049
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Dengue, Yellow Fever, Zika and Chikungunya epidemic arboviruses in Brazil

Debora Ferreira Barreto-Vieira / Dinair Couto-Lima / Fernanda Cunha Jácome / Gabriela Cardoso Caldas / Ortrud Monika Barth

Memórias do Instituto Oswaldo Cruz., Vol

ultrastructural aspects

2021 Volume 115

Abstract: BACKGROUND The impact of arbovirus cocirculation in Brazil is unknown. Dengue virus (DENV) reinfection may result in more intense viraemia or immunopathology, leading to more severe disease. The Zika virus (ZIKV) epidemic in the Americas provided ... ...

Abstract	BACKGROUND The impact of arbovirus cocirculation in Brazil is unknown. Dengue virus (DENV) reinfection may result in more intense viraemia or immunopathology, leading to more severe disease. The Zika virus (ZIKV) epidemic in the Americas provided pathogenicity evidence that had not been previously observed in flavivirus infections. In contrast to other flaviviruses, electron microscopy studies have shown that ZIKV may replicate in viroplasm-like structures. Flaviviruses produce an ensemble of structurally different virions, collectively contributing to tissue tropism and virus dissemination. OBJECTIVES AND METHODS In this work, the Aedes albopictus mosquito cell lineage (C6/36 cells) and kidney epithelial cells from African green monkeys (Vero cells) were infected with samples of the main circulating arboviruses in Brazil [DENV-1, DENV-2, DENV-3, DENV-4, ZIKV, Yellow Fever virus (YFV) and Chikungunya virus (CHIKV)], and ultrastructural studies by transmission electron microscopy were performed. FINDINGS We observed that ZIKV, the DENV serotypes, YFV and CHIKV particles are spherical. ZIKV, DENV-1, -2, -3 and -4 presented diameters of 40-50 nm, and CHIKV presented approximate diameters of 50-60 nm. Viroplasm-like structures was observed in ZIKV replication cycle. MAIN CONCLUSIONS The morphogenesis of these arboviruses is similar to what has been presented in previous studies. However, we understand that further studies are needed to investigate the relationship between viroplasm-like structures and ZIKV replication dynamics.
Keywords	arboviruses ; transmission electron microscopy ; viroplasm ; flaviviruses ; Arctic medicine. Tropical medicine ; RC955-962 ; Microbiology ; QR1-502
Language	English
Publishing date	2021-02-01T00:00:00Z
Publisher	Instituto Oswaldo Cruz, Ministério da Saúde
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Low impact to fixed cell processing aiming transmission electron microscopy.

Barth, Ortrud Monika / Silva, Marcos Alexandre Nunes da / Barreto-Vieira, Debora Ferreira

Memorias do Instituto Oswaldo Cruz

2016 Volume 111, Issue 6, Page(s) 411–413

Abstract: In cell culture, cell structures suffer strong impact due to centrifugation during processing for electron microscope observation. In order to minimise this effect, a new protocol was successfully developed. Using conventional reagents and equipments, it ...

Abstract	In cell culture, cell structures suffer strong impact due to centrifugation during processing for electron microscope observation. In order to minimise this effect, a new protocol was successfully developed. Using conventional reagents and equipments, it took over one week, but cell compression was reduced to none or the lowest deformation possible.
MeSH term(s)	Aedes/ultrastructure ; Aedes/virology ; Animals ; Cell Culture Techniques ; Centrifugation/methods ; Cercopithecus aethiops ; Dengue Virus/ultrastructure ; Fixatives ; Indicators and Reagents ; Microscopy, Electron, Transmission/methods ; Vero Cells/ultrastructure
Chemical Substances	Fixatives ; Indicators and Reagents
Language	English
Publishing date	2016-06-03
Publishing country	Brazil
Document type	Journal Article
ZDB-ID	953293-6
ISSN	1678-8060 ; 0074-0276
ISSN (online)	1678-8060
ISSN	0074-0276
DOI	10.1590/0074-02760150433
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Article ; Online: Comparative analysis of liver involvement caused by two DENV-2 lineages using an immunocompetent murine model.

Jácome, Fernanda Cunha / Caldas, Gabriela Cardoso / Rasinhas, Arthur da Costa / de Almeida, Ana Luisa Teixeira / de Souza, Daniel Dias Coutinho / Paulino, Amanda Carlos / Leonardo, Raphael / Barth, Ortrud Monika / Dos Santos, Flavia Barreto / Barreto-Vieira, Débora Ferreira

Scientific reports

2021 Volume 11, Issue 1, Page(s) 9723

Abstract: Dengue (DEN) is the most prevalent arbovirus among humans, and four billion people live at risk of infection. The clinical manifestations of DEN are variable, and the disease may present subclinically or asymptomatically. A quarter of patients develop ... ...

Abstract	Dengue (DEN) is the most prevalent arbovirus among humans, and four billion people live at risk of infection. The clinical manifestations of DEN are variable, and the disease may present subclinically or asymptomatically. A quarter of patients develop classical dengue (CD) or severe dengue (SD), which is potentially lethal and involves vascular permeability changes, severe hemorrhage and organ damage. The involvement of the liver is a fairly common feature in DEN, and alterations range from asymptomatic elevation of transaminases to acute liver failure. Since its introduction in Brazil in 1990, two strains of Dengue virus (DENV) serotype 2 (DENV-2) have been detected: Lineage I, which is responsible for an outbreak in 1991, and Lineage II, which caused an epidemic greater than the previous one and had a different epidemiological profile. To date, studies on different strains of the same serotype/genotype and their association with disease severity are scarce. In addition, one of the greatest challenges regarding the study of DEN pathogenesis and the development of drug and vaccine therapies is the absence of an animal model that reproduces the disease as it occurs in humans. The main goals of this study were to assess BALB/c mouse susceptibility experimentally infected by two distinct DENV-2 strains and characterize possible differences in the clinical signs and alterations induced in the liver resulting from those infections. Mice infected by the two DENV-2 lineages gained less weight than uninfected mice; however, their livers were slightly heavier. Increased AST and AST levels were observed in infected mice, and the number of platelets increased in the first 72 h of infection and subsequently decreased. Mice infected with both lineages presented leukocytosis but at different times of infection. The histopathological changes induced by both lineages were similar and comparable to the changes observed in DEN fatal cases. The viral genome was detected in two liver samples. The results demonstrate the susceptibility of BALB/c mice to both DENV-2 lineages and suggest that the changes induced by those strains are similar, although for some parameters, they are manifested at different times of infection.
MeSH term(s)	Animals ; Body Temperature ; Body Weight ; Dengue Virus/classification ; Dengue Virus/pathogenicity ; Disease Models, Animal ; Immunocompetence ; Liver/physiopathology ; Liver/virology ; Liver Function Tests ; Mice ; Mice, Inbred BALB C ; Organ Size
Language	English
Publishing date	2021-05-06
Publishing country	England
Document type	Comparative Study ; Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-021-88502-2
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Article: Brazilian Dengue Virus Type 2-Associated Renal Involvement in a Murine Model: Outcomes after Infection by Two Lineages of the Asian/American Genotype.

Jácome, Fernanda Cunha / Caldas, Gabriela Cardoso / Rasinhas, Arthur da Costa / de Almeida, Ana Luisa Teixeira / de Souza, Daniel Dias Coutinho / Paulino, Amanda Carlos / da Silva, Marcos Alexandre Nunes / Barth, Ortrud Monika / Dos Santos, Flávia Barreto / Barreto-Vieira, Debora Ferreira

Pathogens (Basel, Switzerland)

2021 Volume 10, Issue 9

Abstract: Dengue virus type 2 (DENV-2) is, traditionally, the most studied serotype due to its association with explosive outbreaks and severe cases. In Brazil, almost 20 years after the first introduction in the 1990s, a new lineage (Lineage II) of the DENV-2 ... ...

Abstract	Dengue virus type 2 (DENV-2) is, traditionally, the most studied serotype due to its association with explosive outbreaks and severe cases. In Brazil, almost 20 years after the first introduction in the 1990s, a new lineage (Lineage II) of the DENV-2 Asian/American genotype emerged and caused an epidemic with severe cases and hospitalizations. Severe dengue includes multiple organ failure, and renal involvement can be potentially related to increased mortality. In order to better understand the role of DENV infection in renal injury, here we aimed to investigate the outcomes of infection with two distinct lineages of DENV-2 Asian/American genotype in the kidney of a murine model. BALB/c mice were infected with Lineages I and II and tissues were submitted to histopathology, immunohistochemistry, histomorphometry and ultrastructural analysis. Blood urea nitrogen (BUN) was detected in blood sample accessed by cardiac puncture. A tendency in kidney weight increase was observed in mice infected with both lineages, but urea levels, on average, were increased only in mice infected with Lineage II. The DENV antigen was detected in the tissue of mice infected with Lineage II and morphological changes were similar to those observed in human dengue cases. Furthermore, the parameters such as organ weight, urea levels and morphometric analysis, showed significant differences between the two lineages in the infected BALB/c, which was demonstrated to be a suitable experimental model for dengue pathophysiology studies in kidneys.
Language	English
Publishing date	2021-08-26
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2695572-6
ISSN	2076-0817
ISSN	2076-0817
DOI	10.3390/pathogens10091084
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Book: Modelo animal experimental para o estudo da patogênese dos vírus dengue sorotipos 1 e 2

Barreto-Vieira, Débora Ferreira

manual de técnicas

2010

Author's details	Débora Ferreira Barreto-Vieira, Ortrud Monika Barth-Schatzmayr, Hermann Gonçalves Schatzmayr
MeSH term(s)	Densovirus/pathogenicity ; Disease Models, Animal
Language	Portuguese
Size	xiv, 82 pages :, illustrations (chiefly color) ;, 23 cm
Document type	Book
ISBN	9788571932357 ; 8571932352
Database	Catalogue of the US National Library of Medicine (NLM)

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