MUSSYA CISOTTO ROCHA

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LIM/48 - Laboratório de Imunologia, Hospital das Clínicas, Faculdade de Medicina

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Agora exibindo 1 - 10 de 18
  • article 7 Citação(ões) na Scopus
    Oral administration of buparvaquone nanostructured lipid carrier enables in vivo activity against Leishmania infantum
    (2022) MONTEIRO, Lis Marie; LOBENBERG, Raimar; BARBOSA, Eduardo Jose; ARAUJO, Gabriel Lima Barros de; SATO, Paula Keiko; KANASHIRO, Edite; ELIODORO, Raissa H. de Araujo; ROCHA, Mussya; FREITAS, Vera Lucia Teixeira de; FOTAKI, Nikoletta; BOU-CHACRA, Nadia Araci
    Leishmaniasis, a neglected tropical disease, is prevalent in 98 countries with the occurrence of 1.3 million new cases annually. The conventional therapy for visceral leishmaniasis requires hospitalization due to the severe adverse effects of the drugs, which are administered parenterally. Buparvaquone (BPQ) showed in vitro activity against leishmania parasites; nevertheless, it has failed in vivo tests due to its low aqueous solubility. Though, lipid nanoparticles can overcome this holdback. In this study we tested the hypothesis whether BPQ-NLC shows in vivo activity against L. infantum. Two optimized formulations were prepared (V1: 173.9 +/- 1.6 nm, 0.5 mg of BPQ/mL; V2: 232.4 +/- 1.6 nm, 1.3 mg of BPQ/mL), both showed increased solubility up to 73.00-fold, and dissolution up to 83.29%, while for the free drug it was only 2.89%. Cytotoxicity test showed their biocompatibility (CC50 >554.4 mu M). Besides, the V1 dose of 0.3 mg/kg/day for 10 days reduced the parasite burden in 83.4% +/- 18.2% (p <0.05) in the liver. BPQ-NLC showed similar leishmanicidal activity compared to miltefosine. Therefore, BPQ-NLC is a promising addition to the limited therapeutic arsenal suitable for leishmaniasis oral administration treatment.
  • article 0 Citação(ões) na Scopus
    New Allele-Specific Oligonucleotide (ASO) amplifications for Toxoplasma gondii rop18 allele typing: Analysis of 86 human congenital infections in Brazil
    (2023) SANTOS, Emilly Henrique dos; BARREIRA, Gabriel Acca; YAMAMOTO, Lidia; ROCHA, Mussya Cisotto; RODRIGUES, Karen Alessandra; CRUZ, Maria Carolina Pires; KANUNFRE, Kelly Aparecida; OKAY, Thelma Suely
    This study aimed to detect and differentiate Toxoplasma gondii by the allele typing of its polymorphic rop18 gene. For this purpose, a novel genotyping system using allele-specific oligonucleotides (ASOs) was designed, consisting of three ASO pairs. The first and third pairs specifically amplify rop18 allele I and allele III, while the second pair amplify both allele I and II. Genomic DNA from 86 congenital infections was analyzed by ASO-PCRs, successfully typing 82 (95.35%) samples. The remaining 4 samples (4.65%) required sequencing and single nucleotide polymorphism (SNP) analysis of the amplification products. The distribution of samples according to rop18 alleles was: 39.5% of allele III, 38.4% of allele II, 19.8% of mixed rop18 alleles (I/III or II/III), and 2.3% of allele I. The six severely compromised infants exhibited the highest parasite load levels and were infected during the first and early second trimesters of pregnancy. Among these cases, two were associated with rop18 allele I parasites, two with mixed rop18 alleles (I/III), one with allele II, and one with allele III parasites. In conclusion, all severe cases of congenital toxoplasmosis were infected during early pregnancy, but they were not exclusively associated with rop18 allele I parasites, as observed in murine toxoplasmosis. Furthermore, nearly one-fifth of parasites were non-archetypal, exhibiting more than one rop18 allele, indicating a higher genetic diversity of Toxoplasma gondii in this South American sample. Overall, a robust T. gondii rop18 allele typing was developed and suggested that congenital toxoplasmosis in humans involves complex mechanisms beyond the parasite genotype.
  • article 12 Citação(ões) na Scopus
    Biodegradable nanocarriers coated with polymyxin B: Evaluation of leishmanicidal and antibacterial potential
    (2019) COSTA, Juliana Souza Ribeiro; MEDEIROS, Marilia; YAMASHIRO-KANASHIRO, Edite Harumi; ROCHA, Mussya Cisotto; COTRIM, Paulo Cesar; STEPHANO, Marco Antonio; LANCELLOTTI, Marcelo; TAVARES, Guilherme Diniz; OLIVEIRA-NASCIMENTO, Laura
    Most treatments of leishmaniasis require hospitalization and present side effects or parasite resistance; innovations in drug formulation/reposition can overcome these barriers and must be pursued to increase therapeutic alternatives. Therefore, we tested polymyxin B (polB) potential to kill Leishmania amazonensis, adsorbed or not in PBCA nanoparticles (PBCAnp), which could augment polB internalization in infected macrophages. PBCAnps were fabricated by anionic polymerization and analyzed by Dynamic Light Scattering (size, potential), Nanoparticle Tracking Analysis (size/concentration), vertical diffusion cell (release rate), drug incorporation (indirect method, protein determination) and in vitro cell viability. Nanoparticles coated with polB (PBCAnp-polB) presented an adequate size of 261.5 25.9 nm, low PDI and of 1.79 +/- 0.17 mV (stable for 45 days, at least). The 50% drug release from PBCAnp-polB was 6-7 times slower than the free polB, which favors a prolonged and desired release profile. Concerning in vitro evaluations, polB alone reduced in vitro amastigote infection of macrophages (10 g/mL) without complete parasite elimination, even at higher concentrations. This behavior limits its future application to adjuvant leishmanicidal therapy or antimicrobial coating of carriers. The nanocarrier PBCAnp also presented leishmanicidal effect and surpassed polB activity; however, no antimicrobial activity was detected. PolB maintained its activity against E. coli, Pseudomonas and Klebsiella, adding antimicrobial properties to the nanoparticles. Thus, this coated drug delivery system, described for the first time, demonstrated antileishmanial and antimicrobial properties. The bactericidal feature helps with concomitant prevention/treatment of secondary infections that worst ulcers induced by cutaneous L. amazonensis, ultimately ending in disfiguring or disabling lesions. Author summary Cutaneous leishmaniasis form disfiguring lesions that leads to depression and social isolation. Both consequences result often in economic and education loss for communities already short on resources. The actual treatment aims to eliminate the protozoa disease-vector. Yet, it presents undesired effects or parasite resistance, besides the need for intravenous infusions and hospitalization. There is a few topical leishmanicidals but none with prolonged action. As highlighted, no ideal therapy exists; we must keep searching for alternatives, which includes new drugs, vehicles or therapies. For drugs, the most cost-effective way comprises testing approved molecules for other diseases, such as the antimicrobial polymyxin B (polB). Repositioning a drug has the added benefit of known safety profile and drawbacks, shortening and narrowing the research path. In turn, carriers can decrease side-effects and multiple dosing. They shield and/or improve drug targeting, which in this case means delivering inside macrophages infected with Leishmania. An example is poly (n-butyl cyanoacrylate) nanoparticles (PBCAnp), shown as an efficient carrier for targeting an extended drug release. Therefore, this work aimed to evaluate leishmanicidal and adjuvant properties of polB, alone and absorbed onto PBCAnp.
  • article 1 Citação(ões) na Scopus
    Genetic diversity of Trypanosoma cruzi strains isolated from chronic chagasic patients and non-human hosts in the state of Sao Paulo, Brazil
    (2022) SOUZA, Thiago Kury Moreno de; WESTPHALEN, Elizabeth Visone Nunes; WESTPHALEN, Sansao da Rocha; TANIGUCHI, Helena Hilomi; ELIAS, Carlos Roberto; MOTOIE, Gabriela; GAVA, Ricardo; PEREIRA-CHIOCCOLA, Vera Lucia; NOVAES, Christina Terra Gallafrio; CARVALHO, Noemia Barbosa; BOCCHI, Edimar Alcides; CRUZ, Fatima das Dores da; ROCHA, Mussya Cisotto; SHINJO, Samuel Katsuyuki; SHIKANAI-YASUDA, Maria Aparecida; ORTIZ, Paola Andrea; TEIXEIRA, Marta Maria Geraldes; TOLEZANO, Jose Eduardo
    BACKGROUND Trypanosoma cruzi shows an exuberant genetic diversity. Currently, seven phylogenetic lineages, called discrete typing units (DTUs), are recognised: TcI-TcVI and Tcbat. Despite advances in studies on T. cruzi and its populations, there is no consensus regarding its heterogeneity. OBJECTIVES This study aimed to perform molecular characterisation of T. cruzi strains, isolated in the state of S??o Paulo, to identify the DTUs involved and evaluate their genetic diversity. METHODS T. cruzi strains were isolated from biological samples of chronic chagasic patients, marsupials and triatomines through culture techniques and subjected to molecular characterisation using the fluorescent fragment length barcoding (FFLB) technique. Subsequently, the results were correlated with complementary information to enable better discrimination between the identified DTUs. FINDINGS It was possible to identify TcI in two humans and two triatomines; TcII/VI in 19 humans, two marsupials and one triatomine; and TcIII in one human host, an individual that also presented a result for TcI, which indicated the possibility of a mixed infection. Regarding the strains characterised by the TcII/VI profile, the correlation with complementary information allowed to suggest that, in general, these parasite populations indeed correspond to the TcII genotype. MAIN CONCLUSIONS The TcII/VI profile, associated with domestic cycles and patients with chronic Chagas disease, was the most prevalent among the identified DTUs. Furthermore, the correlation of the study results with complementary information made it possible to suggest that TcII is the predominant lineage of this work.
  • article 19 Citação(ões) na Scopus
    APPLICABILITY OF kDNA-PCR FOR ROUTINE DIAGNOSIS OF AMERICAN TEGUMENTARY LEISHMANIASIS IN A TERTIARY REFERENCE HOSPITAL
    (2013) SATOW, Marcela M.; YAMASHIRO-KANASHIRO, Edite H.; ROCHA, Mussya C.; OYAFUSO, Luiza K.; SOLER, Rita C.; COTRIM, Paulo C.; LINDOSO, Jose Angelo L.
    This study evaluated the applicability of kDNA-PCR as a prospective routine diagnosis method for American tegumentary leishmaniasis (ATL) in patients from the Instituto de Infectologia Emolio Ribas (IIER), a reference center for infectious diseases in Sao Paulo - SP, Brazil. The kDNA-PCR method detected Leishmania DNA in 87.5% (112/128) of the clinically suspected ATL patients, while the traditional methods demonstrated the following percentages of positivity: 62.8% (49/78) for the Montenegro skin test, 61.8% (47/76) for direct investigation, and 19.3% (22/114) for in vitro culture. The molecular method was able to confirm the disease in samples considered negative or inconclusive by traditional laboratory methods, contributing to the final clinical diagnosis and therapy of ATL in this hospital. Thus, we strongly recommend the inclusion of kDNA-PCR amplification as an alternative diagnostic method for ATL, suggesting a new algorithm routine to be followed to help the diagnosis and treatment of ATL in IIER.
  • conferenceObject
    PREVENTION OF TRANSFUSIONAL MALARIA IN THE STATE OF SAO PAULO BRAZIL
    (2017) SANTI, Silvia M. Di; SANCHEZ, Maria Carmen A.; MENDRONE JR., Alfredo; LIMA, Giselle F.; ROCHA, Mussya C.; COSTA-NASCIMENTO, Maria J.; FUJIMORI, Mahyumi; LEVI, Jose E.
  • article 5 Citação(ões) na Scopus
    Gastrointestinal manifestations are associated with severe pediatric COVID-19: A study in tertiary hospital
    (2021) PAULA, Camila Sanson Yoshino de; PALANDRI, Giovanna Gavros; FONSECA, Taiane Siraisi; VENDRAMINI, Thais Cristina Annibale; FARHAT, Sylvia Costa Lima; PEREIRA, Maria Fernanda Badue; LITVINOV, Nadia; TOMA, Ricardo Katsuya; SA, Fernanda Viveiros Moreira de; RODRIGUES, Katharina Reichmann; SCHVARTSMAN, Claudio; FORSAIT, Silvana; SAKITA, Neusa Keico; KANUNFRE, Kelly Aparecida; ROCHA, Mussya Cisotto; SANTOS, Emilly Henrique dos; OKAY, Thelma Suely; PINHO, Joao Renato Rebello; CARVALHO, Werther Brunow de; CARNEIRO-SAMPAIO, Magda; SILVA, Clovis Artur Almeida; MARQUES, Heloisa Helena de Sousa; EISENCRAFT, Adriana Pasmanik; ROSSI JUNIOR, Alfio; DELGADO, Artur Figueiredo; LEAL, Gabriela Nunes; FRAMIL, Juliana Valeria de Souza; GIBELLI, Maria Augusta Bento Cicaroni; JORGE, Patricia Palmeira Daenekas
  • article 20 Citação(ões) na Scopus
    Targeting Leishmania amazonensis amastigotes through macrophage internalisation of a hydroxymethylnitrofurazone nanostructured polymeric system
    (2017) MONTEIRO, Lis Marie; LOBENBERG, Raimar; FERREIRA, Elizabeth Igne; COTRIM, Paulo Cesar; KANASHIRO, Edite; ROCHA, Mussya; CHUNG, Man Chin; BOU-CHACRA, Nadia
    Dextran-coated poly (n-butyl cyanoacrylate) nanoparticles (PBCA-NPs) were prepared and were evaluated for enhanced delivery of a promising anti-Leishmania drug candidate, hydroxymethylnitrofurazone (NFOH), to phagocytic cells. Currently available chemotherapy for leishmaniasis, such as pentavalent antimonials, presents low safety and efficacy. Furthermore, widespread drug resistance in leishmaniasis is rapidly emerging. To overcome these drawbacks, the use of nanosized delivery systems can reduce systemic drug toxicity and increase the drug concentration in infected macrophages, therefore improving treatment of leishmaniasis. PBCA-NPs containing NFOH (PBCA-NFOH-NPs) were prepared by an anionic emulsion polymerisation method. The z-average and polydispersity index (PDI) were determined by photon correlation spectroscopy, the zeta potential by microelectrophoresis and the entrapment efficiency by HPLC. Cytotoxicity was determined using macrophages from BALB/c mice. Efficacy tests were performed using Leishmania amazonensis promastigotes and amastigotes. The z-average of PBCA-NFOH-NPs was 151.5 +/- 61.97 nm, with a PDI of 0.104 +/- 0.01, a zeta potential of -10.1 +/- 6.49 mV and an entrapment efficiency of 64.47 +/- 0.43%. Efficacy in amastigotes revealed IC50 values of 0.33 mu M and 31.2 mu M for the nanostructured and free NFOH, respectively (95-fold increase). The cytotoxicity study indicated low toxicity of the PBCA-NFOH-NPs to macrophages. The selectivity index was 370.6, which is 49-fold higher than free NFOH (7.6). Such findings indicated that improved efficacy could be due to NP internalisation following site-specific drug delivery and reactivation of immune protective reactions by the NP components. Thus, PBCA-NFOH-NPs have the potential to significantly improve the treatment of leishmaniasis, with reduced systemic side effects.
  • article 2 Citação(ões) na Scopus
    Silent circulation of Chikungunya virus among pregnant women and newborns in the Western Brazilian Amazon before the first outbreak of chikungunya fever
    (2022) KANUNFRE, Kelly Aparecida; ROCHA, Mussya Cisotto; MALTA, Maira Barreto; SOUZA, Rodrigo Medeiros de; CASTRO, Marcia Caldas; BOSCARDIN, Silvia Beatriz; SOUZA, Higo Fernando Santos; WITKIN, Steven S.; CARDOSO, Marly Augusto; OKAY, Thelma Suely
    The prevalence of immunity to Chikungunya virus (CHIKV) in pregnant women and newborns in the Western Brazilian Amazon was assessed at a time when previous studies did not report chikungunya fever in the area. In 435 asymptomatic pregnant women and 642 healthy unrelated newborns, the presence of IgM and IgG antibodies to CHIKV were determined by a commercial ELISA. All participants were negative to IgM anti-CHIKV. Anti-CHIKV IgG was identified in 41 (9.4%) pregnant women and 66 (10.3%) newborns. The presence of anti-CHIKV IgG was positively associated with the lowest socioeconomic status in pregnant women (OR 2.54, 95% CI 1.15-5.62, p=0.021) and in the newborns' mothers (OR 5.10, 95% CI 2.15-12.09, p< 0.001). Anti-CHIKV IgG was also associated with maternal age in both, the pregnant women (OR 1.06, 95% CI 1.00-1.11, p=0.037) and the newborns'mothers (OR 1.08, 95% CI 1.03-1.12, p=0.001). Pregnancy outcomes in which the mother or the newborn was anti-CHIKV IgG positive proceeded normally. Negative CHIKV serology was associated with being positive for DENV antibodies and having had malaria during pregnancy. These findings showed that there was already a silent circulation of CHIKV in this Amazon region before the first outbreak of chikungunya fever. Furthermore, seropositivity for CHIKV was surprisingly frequent (10%) in both, pregnant women and newborns, affecting mainly low-income women.
  • article 14 Citação(ões) na Scopus
    SARS-CoV-2 infections with emphasis on pediatric patients: a narrative review
    (2020) YAMAMOTO, Lidia; SANTOS, Emilly Henrique dos; PINTO, Lacyane Silva; ROCHA, Mussya Cisotto; KANUNFRE, Kelly Aparecida; VALLADA, Marcelo Genofre; OKAY, Thelma Suely
    This narrative review summarizes the main aspects underlying the new coronavirus SARS-CoV-2, its epidemiology, pathophysiology, pointing to differences of SARS-CoV-2 main receptors ACE2, in terms of expression and the amount of soluble ACE2 in the circulation of children, men and women, and also in those with risk factors such as the smokers and pregnant women or presenting with comorbidities (diabetes, obesity, hypertension and other cardiovascular diseases, renal and CNS pre-existing diseases). Clinical manifestations in adults and children were also described, emphasizing the particularities already seen in children, regarding signs, symptoms, viral excretion time and the involvement of all organs and systems. The COVID-19 in the pediatric population was divided into two sections: one dedicated to previously healthy children and adolescents with COVID-19, and the other to those who live with comorbidities and acquired COVID-19. A few paragraphs were reserved to the recently described severe multisystemic inflammatory syndrome associated with COVID-19 (MIS-C) that shares certain characteristics with Kawasaki disease. Some studies on the infection in pregnant and postpartum women, as well as neonates were shown. This review has also covered the laboratory diagnosis of COVID-19, passing through the imaging diagnosis made by the chest tomography revealing ground glass patching opacities, and results of non-specific exams such as the total blood with lymphopenia, the coagulation tests with increased prothrombin times, as well as marked increments of the D-dimer, troponin and proinflammatory cytokines. In the section devoted to the specific laboratory diagnosis of COVID-19, the most used RT-PCR protocols were described and some studies on the serological diagnosis with IgA, IgM and IgG detection were detailed, including the use of rapid immunochromatographic assays and discussing the ideal period after the onset of symptoms to perform each type of test. In the end, the management of pediatric patients with COVID-19 based mainly on supportive measures has been briefly commented.