Morphological Studies of Hepatic Tissue of Mice Reinfected With Dengue Virus Serotypes 1 or 2

Schatzmayr, Hermann G.; Takiya, Christina Maeda; Jácome, Fernanda Cunha; Silva, Manoel Enderson Vieira; Faria, Nieli Rodrigues da Costa; Nogueira, Rita Maria Ribeiro; Barth, Ortrud Monika

doi:10.17525/vrr.v15i2.68

Cited by 6 publications

(5 citation statements)

References 28 publications

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“…Morphological analysis - Cells were fixed with 1% glutaraldehyde in sodium cacodylate buffer (0.2 M, pH 7.2), post-fixed with 1% buffered osmium tetroxide, dehydrated in acetone, embedded in epoxy resin, and polymerised at 60ºC for three days ( Sesso 2007 , Barreto-Vieira et al 2010 , 2015 ). The resin blocks were then cut into 50-70 nm thick ultrathin sections.…”

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confidence: 99%

Ultrastructure of Zika virus particles in cell cultures

Barth

Silva

Santos

et al. 2016

Mem. Inst. Oswaldo Cruz

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Zika virus (ZIKV) has infected thousands of Brazilian people and spread to other American countries since 2015. The introduction of ZIKV brought a strong impact to public health in Brazil. It is of utmost importance to identify a susceptible cell line that will enable the isolation and identification of the virus from patient samples, viral mass production, and testing of drug and vaccine candidates. Besides real-time reverse transcriptase polymerase chain reaction diagnosis for detecting the viral genome, virus isolation in cell lines was useful in order to study the structure of the viral particle and its behaviour inside cells. Analysis of ZIKV infected cell lines was achieved using transmission electron microscopy (TEM). Blood was obtained from a Brazilian patient during the first days after presenting with signs of the disease, and ZIKV from the patient’s blood was isolated in the C6/36 mosquito cell line. Afterwards, Vero cells were inoculated with the viral suspension, fixed six days after inoculation, embedded in polymers, and ultra-thin cut. Like dengue viruses, this flavivirus showed numerous virus particles present inside cellular vesicles thereby confirming the susceptibility of the Vero cell line to ZIKV replication. TEM is a unique technique available to make the virus visible.

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confidence: 99%

Ultrastructure of Zika virus particles in cell cultures

Barth

Silva

Santos

et al. 2016

Mem. Inst. Oswaldo Cruz

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“…For TEM analyses the Vero-E6 cells suspensions were fixed in 2.5% glutaraldehyde in sodium cacodilate buffer (0.2 M, pH 7.2), post-fixed in 1% buffered osmium tetroxide, dehydrated in acetone, embedded in epoxy resin and polymerised at 60 °C over the course of three days 81 , 82 . Ultrathin sections (50–70 nm) were obtained from the resin blocks.…”

Section: Methodsmentioning

confidence: 99%

Inactivation of SARS-CoV-2 by a chitosan/α-Ag2WO4 composite generated by femtosecond laser irradiation

Silva

Pimentel

Pinatti

et al. 2022

Sci Rep

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In the current COVID-19 pandemic, the next generation of innovative materials with enhanced anti-SARS-CoV-2 activity is urgently needed to prevent the spread of this virus within the community. Herein, we report the synthesis of chitosan/α-Ag2WO4 composites synthetized by femtosecond laser irradiation. The antimicrobial activity against Escherichia coli, Methicilin-susceptible Staphylococcus aureus (MSSA), and Candida albicans was determined by estimating the minimum inhibitory concentration (MIC) and minimal bactericidal/fungicidal concentration (MBC/MFC). To assess the biocompatibility of chitosan/α-Ag2WO4 composites in a range involving MIC and MBC/MFC on keratinocytes cells (NOK-si), an alamarBlue™ assay and an MTT assay were carried out. The SARS-CoV-2 virucidal effects was analyzed in Vero E6 cells through viral titer quantified in cell culture supernatant by PFU/mL assay. Our results showed a very similar antimicrobial activity of chitosan/α-Ag2WO4 3.3 and 6.6, with the last one demonstrating a slightly better action against MSSA. The chitosan/α-Ag2WO4 9.9 showed a wide range of antimicrobial activity (0.49–31.25 µg/mL). The cytotoxicity outcomes by alamarBlue™ revealed that the concentrations of interest (MIC and MBC/MFC) were considered non-cytotoxic to all composites after 72 h of exposure. The Chitosan/α-Ag2WO4 (CS6.6/α-Ag2WO4) composite reduced the SARS-CoV-2 viral titer quantification up to 80% of the controls. Then, our results suggest that these composites are highly efficient materials to kill bacteria (Escherichia coli, Methicillin-susceptible Staphylococcus aureus, and the yeast strain Candida albicans), in addition to inactivating SARS-CoV-2 by contact, through ROS production.

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“…For ultrastructural analysis, the infected and noninfected Calu-3 cell monolayers were trypsinized at 24 and 48 hours postinfection or cultivation, respectively. Cell suspensions were xed in 2.5% glutaraldehyde in sodium cacodylate buffer (0.2 M, pH 7.2), post xed in 1% buffered osmium tetroxide, dehydrated in acetone, embedded in epoxy resin, and polymerized at 60°C over the course of three days 39,40 . Ultrathin sections (50-70 nm) were obtained from the resin blocks.…”

Section: Methodsmentioning

confidence: 99%

SARS-CoV-2 engages replication and inflammasome activation through lipid remodeling via SREBPs

Bozza

Soares

Dias

et al. 2022

Preprint

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SARS-CoV-2 and other ssRNA + viruses induce 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 acid metabolism, while SREBP2 controls cholesterol metabolism, and both isoforms are associated with lipid droplet (LD) biogenesis. SARS-CoV-2 infection has been shown to increase the expression and activation of SREBPs, but the impact of this pathway on the infection outcome is still poorly explored. Here, we evaluated the effect of pharmacologic and molecular inhibition of SREBP1 and SREBP2 in a SARS-CoV-2-infected lung epithelial cell line (Calu-3). We showed that SARS-CoV-2 infection induced the expression and activation of SREBP1 and SREBP2, enzymes of lipid metabolism and LD accumulation. Partial inhibition of SARS-CoV-2 replication and cell death was observed with the genetic knockdown of SREBP1 or SREBP2, while combined SREBP1 and SREBP2 knockdown led to synergistic inhibition. Combined SREBP1 and SREBP2 knockdown inhibited DGAT-1 expression and abrogated SARS-CoV-2-triggered LD formation in Calu-3 cells. Moreover, blockage of LD biogenesis by DGAT1 siRNA inhibited SARS-CoV-2 replication and cell death. Pharmacological inhibition with the dual SREBP activation inhibitor fatostatin reduced virus replication, cell death and LD biogenesis. In addition, we demonstrated that SARS-CoV-2 induced cell death by pyroptosis, with activation of caspase-1, cleavage of gasdermin D1 and release of IL-1β and IL-18 depending on SREBP activation. Collectively, our findings help to elucidate that SREBPs are crucial host factors required for viral replication, LD biogenesis and inflammasome activation and indicate SREBP as a host target for the development of antiviral strategies.

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Morphological Studies of Hepatic Tissue of Mice Reinfected With Dengue Virus Serotypes 1 or 2

Cited by 6 publications

References 28 publications

Ultrastructure of Zika virus particles in cell cultures

Ultrastructure of Zika virus particles in cell cultures

Inactivation of SARS-CoV-2 by a chitosan/α-Ag2WO4 composite generated by femtosecond laser irradiation

SARS-CoV-2 engages replication and inflammasome activation through lipid remodeling via SREBPs

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