VERA LUCIA TEIXEIRA DE FREITAS

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Departamento de Moléstias Infecciosas e Parasitárias, Faculdade de Medicina
LIM/48 - Laboratório de Imunologia, Hospital das Clínicas, Faculdade de Medicina

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  • article 4 Citação(ões) na Scopus
    A Real Time PCR strategy for the detection and quantification of Candida albicans in human blood
    (2020) BUSSER, Felipe Delatorre; COELHO, Vivian Caso; FONSECA, Claudia de Abreu; NEGRO, Gilda Maria Barbaro Del; SHIKANAI-YASUDA, Maria Aparecida; LOPES, Marta Heloisa; MAGRI, Marcello Mihailenko Chaves; FREITAS, Vera Lucia Teixeira de
    Candidemia is a significant cause of bloodstream infections (BSI) in nosocomial settings. The identification of species can potentially improve the quality of care and decrease human mortality. Quantitative PCR (qPCR) was evaluated for Candida albicans detection using culture suspensions containing C. albicans, spiked human blood. the cloned qPCR target fragment (ITS2 region) and the results of these assays were compared. The assays showed a good detection limit: C. albicans DNA extracted from yeast (sensitivity 0.2 CFU/mu L), spiked human blood (sensitivity 10 CFU/mL), and cloned fragment of ITS2 region (sensitivity 20 target copies/mu L). The efficiency of ITS2 fragment-qPCR ranged from 89.67 to 97.07, and the linearity (R-2) of the standard curve ranged from 0.992 to 0.999. The results showed that this ITS2-qPCR has a great potential as a molecular prototype model for the development of a test to be applied in clinical practice, greatly reducing the time of candidemia diagnosis, which is extremely important in this clinical setting.
  • 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.
  • conferenceObject
    CHAGAS DISEASE CARDIOMYOPATHY: ASSOCIATION WITH IL-17 AND IL-18 GENETIC POLYMORPHISMS
    (2018) SANTOS, Alexandra dos; FERREIRA, Daiane; OLIVEIRA, Jamile; OLIVEIRA, Claudia; BOCCHI, Edimar; NOVAES, Cristina; CRUZ, Fatima; CARVALHO, Noemia; SATO, Paula; FREITAS, Vera; SHIKANAI-YASUDA, Maria
  • article 4 Citação(ões) na Scopus
    Suspected vertical transmission of Chagas disease caused by DTU TcIV in an infection probably transmitted orally, during anoutbreak in the Brazilian Amazon
    (2021) FREITAS, Vera Lucia Teixeira de; ESPER, Helena Rangel; NAKANISHI, Erika Shimoda; PIOTTO, Mariana Ramos; ASSY, Joao Guilherme Pontes Lima; BERRETA, Olivia Campos Pinheiro; SAID, Renato do Carmo; SEGURADO, Aluisio Augusto Cotrim; CARVALHO, Noemia Barbosa; FRANCA, Francisco Oscar de Siqueira; LOPES, Marta Heloisa
    This study describes difficulties in the monitoring of a child born during an oral outbreak of Chagas disease, in which there are several indications that the transmission occurred through the congenital route: 1. the mother was in the third trimester of pregnancy when she was infected; 2. She presented high parasitemia at the time of delivery; 3. In both, the mother and her daughter, T cruzi was classified as DTU TcIV. The parasites were not found in the blood at birth and the infection was detected only three months later in an asymptomatic infant. As the mother and her child live in a highly endemic area, vector transmission could not be excluded during this period.
  • article 21 Citação(ões) na Scopus
    Multilocus Sequence Typing of Candida tropicalis Shows the Presence of Different Clonal Clusters and Fluconazole Susceptibility Profiles in Sequential Isolates from Candidemia Patients in Sao Paulo, Brazil
    (2013) MAGRI, Marcello Mihailenko Chaves; GOMES-GOUVEA, Michele Soares; FREITAS, Vera Lucia Teixeira de; MOTTA, Adriana Lopes; MORETTI, Maria Luiza; SHIKANAI-YASUDA, Maria Aparecida
    The profiles of 61 Candida tropicalis isolates from 43 patients (28 adults and 15 children) diagnosed with candidemia at two teaching hospitals in Sao Paulo, Brazil, were characterized by multilocus sequence typing (MLST). For the 14 patients who had bloodstream infections, 32 isolates were serially collected from their blood and/or catheters. Thirty-nine diploid sequence types (DSTs) were differentiated. According to the C. tropicalis MLST database (http://pubmlst.org/ctropicalis/), 36 DSTs and 23 genotypes identified from the 61 isolates had not previously been described. This report represents the first study to characterize sequential isolates of C. tropicalis from candidemia cases in South America. Microvariation in a single gene was found in the sequential isolates from 7 patients. The main polymorphisms occurred in the alleles of the XYR1 gene, specifically at nucleotide positions 215, 242, and 344. Macrovariation in six gene fragments was detected in the isolates from 3 patients. eBURST analysis added two new groups to this study (groups 6 and 18). Additionally, susceptibility tests indicate that 3 isolates were resistant to fluconazole. No correlation was found between the DSTs and susceptibility to fluconazole and/or selective antifungal pressure. Two patients were sequentially infected with resistant and susceptible strains. MLST is an important tool for studying the genetic diversity of multiple/sequential isolates of patients with candidemia, allowing the comparison of our data with those from other regions of the world, as well as allowing an analysis of the genetic relationship among several clones in sequential isolates from the same or different candidemia patient sites (blood or catheter).
  • article 13 Citação(ões) na Scopus
    Polymorphisms on IFNG, IL12B and IL12RB1 genes and paracoccidioidomycosis in the Brazilian population
    (2016) CARVALHO, F. M. C.; BUSSER, F. D.; FREITAS, V. L. T.; FURUCHO, C. R.; SADAHIRO, A.; KONO, A. S. G.; CRIADO, P. R.; MORETTI, M. L.; SATO, P. K.; SHIKANAI-YASUDA, M. A.
    Paracoccidioidomycosis (PCM) is a systemic chronic mycosis, endemic in Latin America, especially Brazil, and is the eighth leading cause of death among chronic and recurrent infectious diseases. PCM infection is characterized by the presence of Th1 immune response; the acute form, by a mixed Th2/Th9, while the chronic form is characterized by Th17/Th22 profiles. The occurrence and severity of human PCM may also be associated with genetic factors such as single nucleotide polymorphisms (SNP) on cytokines encoding genes. We investigated the association between these polymorphisms and the different clinical forms of PCM. We included 156 patients with PCM(40 with the acute form, 99 with the chronic multifocal and 17 with the chronic unifocal form) and assayed their DNA samples for IFNG + 874 T/A SNP by PCR-ARMS (Amplification Refractory Mutational System), IL12B + 1188 A/C SNP on 3' UTR and IL12RB1 641 A/G SNP on exon 7 by PCR-RFLP (Restriction Fragment Length Polymorphism). We found similar genotypic and allelic frequencies of the investigated SNPs among the clinical forms of PCM. Considering male patients, the IL12RB1 641 AA genotype was more frequent in the chronic multifocal form while heterozygosis was in the chronic unifocal form of PCM (p = 0.048). Although our data suggest that the AA genotype (IL12RB1) may be associated with the more disseminated chronic disease, more patients of the chronic unifocal PCM group need to be analyzed as well as the secretion patterns of IFN-gamma combined with the IL-12R beta 1 expression for a better comprehension of this association.
  • conferenceObject
    Interferon c and interleukin-12p40 genetic polymorphisms in brazilian patients with paracoccidioidomycosis
    (2012) HOLANDA, F. M. C.; SATO, P. K.; FREITAS, V. L. T.; YASUDA, M. A. Shikanai
    Paracoccidioidomycosis, caused by the thermal dimorphic fungus Paracoccidioides brasiliensis, is a chronic systemic granulomatous disease, endemic in South America, mainly in Brazil, being the eighth leading cause of death among infectious and parasitic diseases. Paracoccidioidomycosis infection is characterized by a Th1 response with secretion of IFN-c and basal levels of IL-10 and IL-4. In the acute form or juvenile a Th2 profile is predominant, with production of IL-10 and IL-4 and low levels of IFN-c. In chronic or adult, a mixed Th1 and Th2 profile is observed. The IFN-c can be induced by another cytokine, IL-12, and both are important in cellular immune response. In experimental models, IFN-c confers resistance to infection caused by Paracoccidioides brasiliensis and deficiency of IL-12p40 can cause susceptibility to this mycosis. The differential production of these cytokines may be associated with genetic polymorphisms. Objective: To investigate the association between the single nucleotide polymorphisms (SNP) IL-12B 3¢ UTR +1188 A/C and IFNG +874 T/A and the clinical forms of paracoccidioidomycosis. Materials and methods: DNA was obtained from peripheral blood samples from patients with paracoccidioidomycosis and healthy subjects. SNP IL-12B 3¢ UTR +1188 A/C was investigated with PCR-RFLP (PCR-Restriction Fragment Length Polymorphism) and SNP IFNG +874 T/A was investigated with PCR-ARMS (Polymerase Chain Reaction - Amplification Refractory Mutational System). Amplified products were visualized with electrophoresis on agarose gel. Results: In the SNP IFNG +874 T/A, the AA genotype was more frequent in the chronic multifocal and TA genotype in the unifocal. Considering the SNP IL-12B 3¢ UTR +1188 A/C there was a tendency for predominance of AC genotype in patients with chronic form, while the AA genotype was more frequent in acute. Despite this apparent difference among the groups, no statistically significant difference was observed in genotype distribution and alleles A, T and C. Conclusions: In this study there was no association between genetic polymorphisms of IL-12B 3¢ UTR +1188 A/C or IFNG +874 T/A and paracoccidioidomycosis, as well as with different clinical forms.
  • article 5 Citação(ões) na Scopus
    Usefulness of PCR for Trypanosoma cruzi DNA in blood and endomyocardial biopsies for detection of Chagas disease reactivation after heart transplantation: A comparative study
    (2021) BENVENUTI, L. A.; FREITAS, V. L. T.; ROGGéRIO, A.; NISHIYA, A. S.; MANGINI, S.; STRABELLI, T. M. V.
    Background: Chagas disease reactivation (CDR) after heart transplantation is characterized by relapse of the infectious disease with proliferation and dissemination of Trypanosoma cruzi parasites. Serial blood PCR testing is consensually recommended for CDR monitoring, but there is uncertainty about the incremental value in performing the molecular tests in endomyocardial biopsies (EMB). Methods: We compared qualitative and quantitative results of PCR for T cruzi DNA in 62 pairs of blood and EMB collected with a maximum time interval of 7 days, from 34 heart-transplanted, chagasic patients. Results: Blood PCR resulted positive in 39/62 (62.9%) samples, with PL ranging from 0.14 to 1610.73 (median: 3.31). PCR resulted positive in 8/60 (13.3%) EMB, with PL ranging from 2.82 to 1670.55 (median: 65.63). All blood samples which tested negative presented a paired EMB which also tested negative. However, 31/39 (79.5%) blood samples which tested positive presented a paired EMB which tested negative. There was poor agreement between blood and EMB PCR (kappa = 0.153). CDR affecting the myocardium (myo-CDR) was diagnosed in three occasions. PCR resulted positive in both blood and EMB at the time of myo-CDR, with PL ranging from 0.61 to 1610.73 in blood and 13.8 to 1670.55 in EMB. Conclusions: Negative PCR for T cruzi in blood rules out myo-CDR, with no value of testing EMB. Positive PCR in blood with high PL is diagnostic for myo-CDR. If PCR in blood results positive with low PL, testing EMB is useful: negative PCR turns unlikely, and positive PCR reinforces greatly the possibility of myo-CDR. © 2021 Wiley Periodicals LLC
  • article 4 Citação(ões) na Scopus
    Detection of Trypanosoma cruzi DTUs TcI and TcIV in two outbreaks of orally-transmitted Chagas disease in the Northern region of Brazil
    (2023) FREITAS, Vera Lucia Teixeira de; PIOTTO, Mariana Ramos; ESPER, Helena Rangel; NAKANISHI, Erika Yoshie Shimoda; FONSECA, Claudia de Abreu; ASSY, Joao Guilherme Pontes Lima; BERRETA, Olivia Campos Pinheiro; FRANCA, Francisco Oscar de Siqueira; LOPES, Marta Heloisa
    This study describes the laboratory investigation of two acute Chagas disease outbreaks that occurred in the riverside communities of Marimarituba and Cachoeira do Arua, in the Santarem municipality, Para State, located in the Northern region of Brazil, and occurred in March 2016 and August 2017, respectively. The generation of data regarding the diversity of Trypanosoma cruzi parasites circulating in the Amazon region is key for understanding the emergence and expansion of Chagas disease. This study aimed to identify T. cruzi Discrete Typing Units (DTUs) involved in two outbreaks of acute Chagas disease (ACD) directly from the patient's biological sample. Nested and multiplex PCR targeting the symbolscript (rRNA) and mini-exon genes, respectively, were used to identify T. cruzi DTU in blood samples from patients diagnosed with ACD. The samples with positive cPCR were submitted for analysis for T. cruzi DTUs, which included 13 samples from the patients with ACD by oral transmission and two samples collected from two newborns of two women with ACD, from Marimarituba and Cachoeira do Arua. The samples were classified as T. cruzi TcIV, from Marimarituba's outbreak, and T. cruzi TcI, from Cachoeira do Arua's outbreak. The molecular identification of T. cruzi may increase understanding of the role of this parasite in Chagas disease's emergence within the Amazon region, contributing to the improvement of the management of this important, but also neglected, disease.
  • article 7 Citação(ões) na Scopus
    Interdisciplinary approach at the primary healthcare level for Bolivian immigrants with Chagas disease in the city of Sao Paulo
    (2017) YASUDA, Maria Aparecida Shikanai; SATOLO, Camila Goncalves; CARVALHO, Noemia Barbosa; ATALA, Magda Maya; FERRUFINO, Rosarlo Quiroga; LEITE, Ruth Moreira; FURUCHO, Celia Regina; LUNA, Expedito; SILVA, Rubens Antonio; HAGE, Marcia; OLIVEIRA, Caroline Medeji Ramos de; BUSSER, Felipe Delatorre; FREITAS, Vera Lucia Teixeira de; WANDERLEY, Dalva Marli Valerio; MARTINELLI, Luzia; ALMEIDA, Sonia Regina; VINAS, Pedro Albajar; CARNEIRO JR., Nivaldo
    Background/Methods In a pioneering cross-sectional study among Bolivian immigrants in the city of Sao Paulo, Brazil, the epidemiological profile, clinical manifestations and morbidity of Chagas disease were described. The feasibility of the management of Chagas disease at primary healthcare clinics using a biomedical and psychosocial interdisciplinary approach was also tested. Previously, a Trypanosoma cruzi (T. cruzi) infection rate of 4.4% among 633 immigrants was reported. The samples were screened using two commercial enzyme-linked immunoassay (ELISA) tests generated with epimastigote antigens, and those with discrepant or seropositive results were analyzed by confirmatory tests: indirect immunofluorescence (IFI), TESA-blot and a commercial recombinant ELISA. PCR and blood cultures were performed in seropositive patients. Results The majority of the 28 seropositive patients were women, of whom 88.89% were of childbearing age. The predominant clinical forms of Chagas disease were the indeterminate and atypical cardiac forms. Less than 50% received the recommended antiparasitic treatment of benznidazole. An interdisciplinary team was centered on primary healthcare physicians who applied guidelines for the management of patients. Infectologists, cardiologists, pediatricians and other specialists acted as reference professionals. Confirmatory serology and molecular biology tests, as well as echocardiography, Holter and other tests, were performed for the assessment of affected organs in secondary healthcare centers. The published high performance of two commercial ELISA tests was not confirmed. Conclusion An interdisciplinary approach including antiparasitic treatment is feasible at the primary healthcare level for the management of Chagas disease in Bolivian immigrants. The itinerant feature of immigration was associated with a lack of adherence to antiparasitic treatment and was considered a main challenge for the clinical management of this population. This approach is recommended for management of the infected population in endemic and nonendemic areas, although different strategies are needed depending on the severity of the disease and the structure of the healthcare system.