Navegando por Autor "Miyazaki, Carolina Kei"
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Item Comparing the therapeutic efficacy of different amphotericin Bcarrying delivery systems against visceral leishmaniasis.(2018) Mendonça, Débora Vasconcelos Costa; Martins, Vivian Tamietti; Lage, Daniela Pagliara; Ribeiro, Patrícia Aparecida Fernandes; Carvalho, Ana Maria Ravena Severino; Dias, Anna Leticia Teotonio; Miyazaki, Carolina Kei; Souza, Daniel Menezes; Roatt, Bruno Mendes; Tavares, Carlos Alberto Pereira; Duarte, Mariana Costa; Coelho, Eduardo Antônio FerrazAmphotericin B (Amp) has been well-successfully used to treat against Leishmania infection, although high toxicity has been found in patients. In the present study, Amp was administered in Leishmania infantum-infected BALB/c mice by three distinct delivery systems aiming to compare their efficacy against challenge infection, as well as their side effects in a murine visceral leishmaniasis (VL) model. This product was administered in a Poloxamer P407 (Pluronic® F127)-based polymeric micelle system (Amp/M), in the Ambisome® formulation (Lip-Amp) or in a free format (free Amp). Glucantime® (Gluc) was used as a comparative drug. Aiming to evaluate different endpoints of the treatments, the efficacy of the compounds was investigated one and 15-days after the therapeutic regimens, determining the parasite load by a limiting dilution assay and a quantitative PCR (qPCR) technique, as well as evaluating the immune response generated in the infected and treated animals. In the results, Amp/M or Lip-Amp-treated mice presented the best outcomes, since significant parasite load reductions were found in the evaluated organs, as well as a parasite-specific Th1 immune response was observed in the animals. In addition, no hepatic or renal damage was found in these mice. On the other hand, free Amp or Gluc induced toxicity in the animals, which was associated with a low Th1 immune response. Comparatively, Amp/M was the most effective drug in our experimental model, and results showed that the Amp-carrying system could be considered as a future alternative in studies against VL.Item Immunodiagnosis of human and canine visceral leishmaniasis using recombinant Leishmania infantum Prohibitin protein and a synthetic peptide containing its conformational B-cell epitope.(2019) Rodrigues, Marcella Rezende; Santos, Lucas Magno Oliveira; Miyazaki, Carolina Kei; Martins, Vivian Tamietti; Ribeiro, Fernanda Ludolf; Kursancew, Amanda Christine da Silva; Ramos, Fernanda Fonseca; Dias, Daniel Silva; Oliveira, Jamil Silvano de; Vieira, Paula Melo de Abreu; Roatt, Bruno Mendes; Ávila, Ricardo Andrez Machado de; Gonçalves, Denise Utsch; Souza, Daniel Menezes; Coelho, Eduardo Antônio Ferraz; Duarte, Mariana CostaIn the present study, Leishmania infantum's Prohibitin was cloned and, alongside a synthetic peptide, evaluated for the serodiagnosis of visceral and tegumentary leishmaniasis (CVL and TL, respectively) in dogs and humans. For TL diagnosis, this study analyzed serum samples from cutaneous (n=20) or mucosal (n=39) leishmaniasis patients, and from Chagas disease (CD) patients (n=8) and non-infected patients (n=45). For CVL diagnosis, serum samples from asymptomatic (n=14), symptomatic (n=71), non-infected (n=116), and Leish-Tec®- vaccinated (n=79) dogs were examined, as well as T. cruzi (n=11) and Ehrlichia canis (n=10) infected animals. An indirect ELISA method using rProhibitin showed diagnostic sensitivity and specificity values of 91.76% and 89.91%, respectively. L. infantum SLA showed 86.11% and 48.24% of specificity and sensitivity, respectively, for CVL serodiagnosis, and 98.31% and 84.91% sensitivity and specificity, respectively for TL diagnosis. L. braziliensis SLA showed 75.47% and 83.05% of specificity and sensitivity, respectively, for TL diagnosis. The synthetic peptide showed a better result in TL than in CVL diagnosis. In conclusion, preliminar results suggest that the detection of antibodies against the rProhibitin protein and the synthetic peptide improves the serodiagnosis of TL and CVL.Item A Pluronic® F127-based polymeric micelle system containing an antileishmanial molecule is immunotherapeutic and effective in the treatment against Leishmania amazonensis infection.(2019) Tavares, Grasiele de Sousa Vieira; Mendonça, Débora Vasconcelos Costa; Miyazaki, Carolina Kei; Lage, Daniela Pagliara; Soyer, Tauane Gonçalves; Carvalho, Lívia Mendes; Ottoni, Flaviano Melo; Dias, Daniel Silva; Ribeiro, Patrícia Aparecida Fernandes; Antinarelli, Luciana Maria Ribeiro; Ribeiro, Fernanda Ludolf; Duarte, Mariana Costa; Coimbra, Elaine Soares; Chávez Fumagalli, Miguel Angel; Roatt, Bruno Mendes; Souza, Daniel Menezes; Barichello, José Mario; Alves, Ricardo José; Coelho, Eduardo Antônio FerrazClioquinol (5-chloro-7-iodoquinolin-8-ol or ICHQ) was recently showed to presents an in vitro effective antileishmanial action, causing changes in membrane permeability, mitochondrial functionality, and parasite morphology. In the present study, ICHQ was incorporated into a Poloxamer 407-based polymeric micelles system (ICHQ/M), and its antileishmanial activity was in vivo evaluated in L. amazonensis-infected BALB/c mice. Amphotericin B (AmpB) and its liposomal formulation (Ambisome®) were used as controls. Parasitological and immunological evaluations were performed 30 days after the treatment. Results indicated more significant reductions in the average lesion diameter and parasite burden in ICHQ or ICHQ/M-treated mice, which were associated with the development of a polarized Th1 immune response, based on production of high levels of IFN-γ, IL-12, TNF-α, GM-CSF, and antileishmanial IgG2a antibody. Control groups´ mice produced high levels of IL-4, IL-10, and IgG1 isotype antibody. No organic toxicity was found by using ICHQ or ICHQ/M to treat the animals, although those receiving AmpB and Ambisome® have presented higher levels of renal and hepatic damage markers. In conclusion, results suggested that the ICHQ/M composition can be considered as an antileishmanial candidate to be tested against human leishmaniasis.