BIANCA HELENA VENTURA FERNANDES
Projetos de Pesquisa
Unidades Organizacionais
SCLAB-05, Faculdade de Medicina
LIM/42 - Laboratório de Hormônios e Genética Molecular, Hospital das Clínicas, Faculdade de Medicina
LIM/42 - Laboratório de Hormônios e Genética Molecular, Hospital das Clínicas, Faculdade de Medicina
4 resultados
Resultados de Busca
Agora exibindo 1 - 4 de 4
- Rederivation of a mutant line (prop 1) of zebrafish Danio rerio infected with Pseudoloma neurophilia using in vitro fertilization with eggs from pathogen-free wild-type (AB) females and sperm from prop 1 males(2022) FERNANDES, Bianca H. Ventura; SILVA, Caroline Caetano da; BISSEGATO, Debora; KENT, Michael L.; CARVALHO, Luciani R.Along with the growing number of laboratories that work with zebrafish (Danio rerio), it is necessary to have animals with good sanitary quality. Specific pathogens can interfere with the experimental results and in the life quality of the animals. Pseudoloma neurophilia is a parasite with high potential for interference in behavioural, morphology, toxicological and genetic research, and is very common in zebrafish facilities. With that, we implemented a protocol for the pathogen elimination in a genetically modified lineage (prop 1) using eggs from specific pathogen-free (SPF) wild-type fish (AB line) for in vitro fertilization, along with water recirculation equipment disinfection, appropriate PCR screening and back crossing protocols. This resulted in SPF prop 1 heterozygotes, which allowed us to move forward with subsequent crossings to develop homozygote prop 1 mutants for our research. Hence, this demonstrates a useful strategy for an individual research laboratory to rederive a specific mutant free line that is not available from other SPF laboratories.
- A novel insight on SARS-CoV-2 S-derived fragments in the control of the host immunity(2023) BASTOS, Thais Sibioni Berti; PAULA, Andre Guilherme Portela de; LUZ, Rebeca Bosso dos Santos; GARNIQUE, Anali M. B.; BELO, Marco A. A.; ETO, Silas Fernandes; FERNANDES, Dayanne Carla; FERRARIS, Fausto Klabund; PONTES, Leticia Gomes de; FRANCA, Tabata Takahashi; BARCELLOS, Leonardo Jose Gil; VERAS, Flavio P.; BERMEJO, Pamela; GUIDELLI, Giovanna; MANEIRA, Carla; MELLO, Fellipe da Silveira Bezerra de; TEIXEIRA, Gleidson; PEREIRA, Goncalo Amarante Guimaraes; FERNANDES, Bianca H. Ventura; SANCHES, Paulo R. S.; BRAZ, Helyson Lucas Bezerra; JORGE, Roberta Jeane Bezerra; MALAFAIA, Guilherme; CILLI, Eduardo M.; OLIVIER, Danilo da Silva; AMARAL, Marcos Serrou do; MEDEIROS, Renata J.; CONDINO-NETO, Antonio; CARVALHO, Luciani R.; MACHADO-SANTELLI, Glaucia M.; CHARLIE-SILVA, Ives; GALINDO-VILLEGAS, Jorge; BRAGA, Tarcio TeodoroDespite all efforts to combat the pandemic of COVID-19, we are still living with high numbers of infected persons, an overburdened health care system, and the lack of an effective and definitive treatment. Understanding the pathophysiology of the disease is crucial for the development of new technologies and therapies for the best clinical management of patients. Since the manipulation of the whole virus requires a structure with an adequate level of biosafety, the development of alternative technologies, such as the synthesis of peptides from viral proteins, is a possible solution to circumvent this problem. In addition, the use and validation of animal models is of extreme importance to screen new drugs and to compress the organism's response to the disease. Peptides derived from recombinant S protein from SARS-CoV-2 were synthesized and validated by in silico, in vitro and in vivo methodologies. Macrophages and neutrophils were challenged with the peptides and the production of inflammatory mediators and activation profile were evaluated. These peptides were also inoculated into the swim bladder of transgenic zebrafish larvae at 6 days post fertilization (dpf) to mimic the inflammatory process triggered by the virus, which was evaluated by confocal microscopy. In addition, toxicity and oxidative stress assays were also developed. In silico and molecular dynamics assays revealed that the peptides bind to the ACE2 receptor stably and interact with receptors and adhesion molecules, such as MHC and TCR, from humans and zebrafish. Macrophages stimulated with one of the peptides showed increased production of NO, TNF-alpha and CXCL2. Inoculation of the peptides in zebrafish larvae triggered an inflammatory process marked by macrophage recruitment and increased mortality, as well as histopathological changes, similarly to what is observed in individuals with COVID-19. The use of peptides is a valuable alternative for the study of host immune response in the context of COVID-19. The use of zebrafish as an animal model also proved to be appropriate and effective in evaluating the inflammatory process, comparable to humans.
- Photobiomodulation Reduces the Cytokine Storm Syndrome Associated with COVID-19 in the Zebrafish Model(2023) ROSA, Ivana F.; PECANHA, Ana P. B.; CARVALHO, Tabata R. B.; ALEXANDRE, Leonardo S.; FERREIRA, Vinicius G.; DORETTO, Lucas B.; SOUZA, Beatriz M.; NAKAJIMA, Rafael T.; SILVA, Patrick da; BARBOSA, Ana P.; GOMES-DE-PONTES, Leticia; BOMFIM, Camila G.; MACHADO-SANTELLI, Glaucia M.; CONDINO-NETO, Antonio; GUZZO, Cristiane R.; PERON, Jean P. S.; ANDRADE-SILVA, Magaiver; CAMARA, Niels O. S.; GARNIQUE, Anali M. B.; MEDEIROS, Renata J.; FERRARIS, Fausto K.; BARCELLOS, Leonardo J. G.; CORREIA-JUNIOR, Jose D.; GALINDO-VILLEGAS, Jorge; MACHADO, Monica F. R.; CASTOLDI, Angela; OLIVEIRA, Susana L.; COSTA, Camila C.; BELO, Marco A. A.; GALDINO, Giovane; SGRO, German G.; BUENO, Natalia F.; ETO, Silas F.; VERAS, Flavio P.; FERNANDES, Bianca H. V.; SANCHES, Paulo R. S.; CILLI, Eduardo M.; MALAFAIA, Guilherme; NOBREGA, Rafael H.; GARCEZ, Aguinaldo S.; CARRILHO, Emanuel; CHARLIE-SILVA, IvesAlthough the exact mechanism of the pathogenesis of coronavirus SARS-CoV-2 (COVID-19) is not fully understood, oxidative stress and the release of pro-inflammatory cytokines have been highlighted as playing a vital role in the pathogenesis of the disease. In this sense, alternative treatments are needed to reduce the level of inflammation caused by COVID-19. Therefore, this study aimed to investigate the potential effect of red photobiomodulation (PBM) as an attractive therapy to downregulate the cytokine storm caused by COVID-19 in a zebrafish model. RT-qPCR analyses and protein-protein interaction prediction among SARS-CoV-2 and Danio rerio proteins showed that recombinant Spike protein (rSpike) was responsible for generating systemic inflammatory processes with significantly increased levels of pro-inflammatory (il1b, il6, tnfa, and nfkbiab), oxidative stress (romo1) and energy metabolism (slc2a1a and coa1) mRNA markers, with a pattern similar to those observed in COVID-19 cases in humans. On the other hand, PBM treatment was able to decrease the mRNA levels of these pro-inflammatory and oxidative stress markers compared with rSpike in various tissues, promoting an anti-inflammatory response. Conversely, PBM promotes cellular and tissue repair of injured tissues and significantly increases the survival rate of rSpike-inoculated individuals. Additionally, metabolomics analysis showed that the most-impacted metabolic pathways between PBM and the rSpike treated groups were related to steroid metabolism, immune system, and lipid metabolism. Together, our findings suggest that the inflammatory process is an incisive feature of COVID-19 and red PBM can be used as a novel therapeutic agent for COVID-19 by regulating the inflammatory response. Nevertheless, the need for more clinical trials remains, and there is a significant gap to overcome before clinical trials can commence.
- Virulomic Analysis of Multidrug-Resistant Klebsiella pneumoniae Isolates and Experimental Virulence Model Using Danio rerio (Zebrafish)(2022) DUARTE, Edson Luiz Tarsia; RIZEK, Camila Fonseca; ESPINOZA, Evelyn Sanchez; MARCHI, Ana Paula; NOGUERA, Saidy Vasconez; CORTES, Marina Farrel; FERNANDES, Bianca H. Ventura; GUIMARAES, Thais; CARRILHO, Claudia M. D. de Maio; V, Lauro Perdigao Neto; TRINDADE, Priscila A.; COSTA, Silvia FigueiredoThis study evaluates a possible correlation between multidrug-resistant Klebsiella pneumoniae strains and virulence markers in a Danio rerio (zebrafish) model. Whole-genome sequencing (WGS) was performed on 46 strains from three Brazilian hospitals. All of the isolates were colistin-resistant and harbored bla(KPC-2). Ten different sequence types (STs) were found; 63% belonged to CC258, 22% to ST340, and 11% to ST16. The virulence factors most frequently found were type 3 fimbriae, siderophores, capsule regulators, and RND efflux-pumps. Six strains were selected for a time-kill experiment in zebrafish embryos: infection by ST16 was associated with a significantly higher mortality rate when compared to non-ST16 strains (52% vs. 29%, p = 0.002). Among the STs, the distribution of virulence factors did not differ significantly except for ST23, which harbored a greater variety of factors than other STs but was not related to a higher mortality rate in zebrafish. Although several virulence factors are described in K. pneumoniae, our study found ST16 to be the only significant predictor of a virulent phenotype in an animal model. Further research is needed to fully understand the correlation between virulence and sequence types.