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LIM/52 - Laboratório de Virologia, Hospital das Clínicas, Faculdade de Medicina

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  • article 0 Citação(ões) na Scopus
    Respiratory viruses and postoperative hemodynamics in patients with unrestrictive congenital cardiac communications: a prospective cohort study
    (2023) ABUD, Kelly C. O.; MACHADO, Clarisse M.; BOAS, Lucy S. Vilas S.; MAEDA, Nair Y.; CARVALHO, Eloisa S.; SOUZA, Maria Francilene S.; GAIOLLA, Paula V.; CASTRO, Claudia R. P.; PEREIRA, Juliana; RABINOVITCH, Marlene; LOPES, Antonio Augusto
    BackgroundPulmonary vascular abnormalities pose a risk for severe life-threatening hemodynamic disturbances following surgical repair of congenital cardiac communications (CCCs). In the distal lung, small airways and vessels share a common microenvironment, where biological crosstalks take place. Because respiratory cells infected by viruses express a number of molecules with potential impact on airway and vascular remodeling, we decided to test the hypothesis that CCC patients carrying viral genomes in the airways might be at a higher risk for pulmonary (and systemic) hemodynamic disturbances postoperatively.MethodsSixty patients were prospectively enrolled (age 11 [7-16] months, median with interquartile range). Preoperative pulmonary/systemic mean arterial pressure ratio (PAP/SAP) was 0.78 (0.63-0.88). The presence or absence of genetic material for respiratory viruses in nasopharyngeal and tracheal aspirates was investigated preoperatively in the absence of respiratory symptoms using real-time polymerase chain reaction (kit for detection of 19 pathogens). Post-cardiopulmonary bypass (CPB) inflammatory reaction was analyzed by measuring serum levels of 36 inflammatory proteins (immunoblotting) 4 h after its termination. Postoperative hemodynamics was assessed using continuous recording of PAP and SAP with calculation of PAP/SAP ratio.ResultsViral genomes were detected in nasopharynx and the trachea in 64% and 38% of patients, respectively. Rhinovirus was the most prevalent agent. The presence of viral genomes in the trachea was associated with an upward shift of postoperative PAP curve (p = 0.011) with a PAP/SAP of 0.44 (0.36-0.50) in patients who were positive versus 0.34 (0.30-0.45) in those who were negative (p = 0.008). The presence or absence of viral genomes in nasopharynx did not help predict postoperative hemodynamics. Postoperative PAP/SAP was positively correlated with post-CPB levels of interleukin-1 receptor antagonist (p = 0.026), macrophage migration inhibitory factor (p = 0.019) and monocyte chemoattractant protein-1 (p = 0.031), particularly in patients with virus-positive tracheal aspirates.ConclusionsPatients with CCCs carrying respiratory viral genomes in lower airways are at a higher risk for postoperative pulmonary hypertension, thus deserving special attention and care. Preoperative exposure to respiratory viruses and post-CPB inflammatory reaction seem to play a combined role in determining the postoperative behavior of the pulmonary circulation.
  • article 64 Citação(ões) na Scopus
    Microbial Translocation Is Associated with Extensive Immune Activation in Dengue Virus Infected Patients with Severe Disease
    (2013) WEG, Cornelia A. M. van de; PANNUTI, Claudio S.; ARAUJO, Evaldo S. A. de; HAM, Henk-Jan van den; ANDEWEG, Arno C.; BOAS, Lucy S. V.; FELIX, Alvina C.; CARVALHO, Karina I.; MATOS, Andreia M. de; LEVI, Jose E.; ROMANO, Camila M.; CENTRONE, Cristiane C.; RODRIGUES, Celia L. de Lima; LUNA, Expedito; GORP, Eric C. M. van; OSTERHAUS, Albert D. M. E.; MARTINA, Byron E. E.; KALLAS, Esper G.
    Background: Severe dengue virus (DENV) disease is associated with extensive immune activation, characterized by a cytokine storm. Previously, elevated lipopolysaccharide (LPS) levels in dengue were found to correlate with clinical disease severity. In the present cross-sectional study we identified markers of microbial translocation and immune activation, which are associated with severe manifestations of DENV infection. Methods: Serum samples from DENV-infected patients were collected during the outbreak in 2010 in the State of Sao Paulo, Brazil. Levels of LPS, lipopolysaccharide binding protein (LBP), soluble CD14 (sCD14) and IgM and IgG endotoxin core antibodies were determined by ELISA. Thirty cytokines were quantified using a multiplex luminex system. Patients were classified according to the 2009 WHO classification and the occurrence of plasma leakage/shock and hemorrhage. Moreover, a (non-supervised) cluster analysis based on the expression of the quantified cytokines was applied to identify groups of patients with similar cytokine profiles. Markers of microbial translocation were linked to groups with similar clinical disease severity and clusters with similar cytokine profiles. Results: Cluster analysis indicated that LPS levels were significantly increased in patients with a profound pro-inflammatory cytokine profile. LBP and sCD14 showed significantly increased levels in patients with severe disease in the clinical classification and in patients with severe inflammation in the cluster analysis. With both the clinical classification and the cluster analysis, levels of IL-6, IL-8, sIL-2R, MCP-1, RANTES, HGF, G-CSF and EGF were associated with severe disease. Conclusions: The present study provides evidence that both microbial translocation and extensive immune activation occur during severe DENV infection and may play an important role in the pathogenesis.
  • article 42 Citação(ões) na Scopus
    Concentration of cytokines in patients with osteoarthritis of the knee and fibromyalgia
    (2014) IMAMURA, Marta; TARGINO, Rosa Alves; HSING, Wu Tu; IMAMURA, Satiko; AZEVEDO, Raymundo Soares; BOAS, Lucy Santos Villas; TOZETTO-MENDOZA, Tania Regina; ALFIERI, Fabio Marcon; FILIPPO, Thais Raquel; BATTISTELLA, Linamara Rizzo
    Introduction: Fibromyalgia and osteoarthritis may present a relationship with the -concentration of cytokines. The aim of this study was to compare the serum concentrations of IL-12p70, tumor necrosis factor, IL-10, IL-6, IL-1 beta, and IL-8 in patients with knee osteoarthritis and fibromyalgia. Materials and methods: The study included 53 women (71.2 +/- 7.6 years old) diagnosed with knee osteoarthritis with moderate-to-severe pain (visual analog scale. 4) for at least 3 months. Sixty women (54.1 +/- 8.1 years old) diagnosed with fibromyalgia according to the American College of Rheumatology criteria and with moderate-to-severe pain (visual analog scale. 4) also participated in this study. For the dosage of cytokines, blood was collected in the morning: 5 mL from the cubital vein. The material was centrifuged at 4 degrees C, separated into 100 mu L aliquots and stored at -80 degrees C until processing. Serum concentrations of the studied cytokines were assessed using the BD Cytometric Bead Array method. Data were analyzed with Student's t-test and the Mann-Whitney U test. Results: We found higher levels of IL-6, IL-10, and IL-1 beta in fibromyalgia patients. After adjustment of age as a covariate, there was no statistically significant difference in the concentration of any cytokine between fibromyalgia and knee osteoarthritis patients. Conclusion: Patients with knee osteoarthritis and fibromyalgia with the same duration and intensity of pain demonstrate similar concentrations of cytokines. Aging may play a role in cytokine profile, a finding not so extensively addressed in the literature and one that should be further investigated.
  • article 0 Citação(ões) na Scopus
    Cytomegalovirus Infection in Inflammatory Bowel Disease Is Not Associated with Worsening of Intestinal Inflammatory Activity (vol 9, e111574, 2014)
    (2015) CARMO, Alexandre Medeiros do; SANTOS, Fabiana Maria; ORTIZ-AGOSTINHO, Carmen Lucia; NISHITOKUKADO, Ieda; FROTA, Cintia S.; GOMES, Flavia Ubeda; LEITE, Andre Zonetti de Arruda; PANNUTI, Claudio Sergio; BOAS, Lucy Santos Vilas; TEIXEIRA, Magaly Gemio; SIPAHI, Aytan Miranda
  • conferenceObject
    SARS-CoV-2 Aerosol Generation During Respiratory Equipment Reprocessing
    (2021) BRUNA, Camila Quartim de Moraes; CIOFI-SILVA, Caroline; GRAZIANO, Kazuko Uchikawa; MENDES-CORREA, Maria Cassia; BOAS, Lucy Santos Villas; FERREIRA, Noeli Evangelista; TOZETTO-MENDOZA, Tania; PAULA, Anderson Vicente de
  • article 23 Citação(ões) na Scopus
    Cytomegalovirus Infection in Inflammatory Bowel Disease Is Not Associated with Worsening of Intestinal Inflammatory Activity
    (2014) CARMO, Alexandre Medeiros do; SANTOS, Fabiana Maria; ORTIZ-AGOSTINHO, Carmen Lucia; NISHITOKUKADO, Ieda; FROTA, Cintia S.; GOMES, Flavia Ubeda; LEITE, Andre Zonetti de Arruda; PANNUTI, Claudio Sergio; BOAS, Lucy Santos Vilas; TEIXEIRA, Magaly Gemio; SIPAHI, Aytan Miranda
    Background: Cytomegalovirus is highly prevalent virus and usually occurs in immunocompromised patients. The pathophysiology and treatment of inflammatory bowel disease often induce a state of immunosuppression. Because this, there are still doubts and controversies about the relationship between inflammatory bowel disease and cytomegalovirus. Aim: Evaluate the frequency of cytomegalovirus in patients with inflammatory bowel disease and identify correlations. Methods: Patients with inflammatory bowel disease underwent an interview, review of records and collection of blood and fecal samples. The search for cytomegalovirus was performed by IgG and IgM blood serology, by real-time PCR in the blood and by qualitative PCR in feces. Results were correlated with red blood cell levels, C-reactive protein levels, erythrocyte sedimentation rates and fecal calprotectin levels for each patient. Results: Among the 400 eligible patients, 249 had Crohn's disease, and 151 had ulcerative colitis. In the group of Crohn's disease, 67 of the patients had moderate or severe disease, but 126 patients presented with active disease, based on the evaluation of the fecal calprotectin. In patients with ulcerative colitis, only 21 patients had moderate disease, but 76 patients presented with active disease, based on the evaluation of the fecal calprotectin. A large majority of patients had positive CMV IgG. Overall, 10 patients had positive CMV IgM, and 9 patients had a positive qualitative detection of CMV DNA by PCR in the feces. All 400 patients returned negative results after the quantitative detection of CMV DNA in blood by real-time PCR. Analyzing the 19 patients with active infections, we only found that such an association occurred with the use of combined therapy (anti-TNF-alpha + azathioprine) Conclusion: The findings show that latent cytomegalovirus infections are frequent and active cytomegalovirus infection is rare. We did not find any association between an active infection of CMV and inflammatory bowel disease activity.
  • article 11 Citação(ões) na Scopus
    Dengue Virus Serotype 2 Intrahost Diversity in Patients with Different Clinical Outcomes
    (2021) TORRES, Maria Celeste; MENDONCA, Marcos Cesar Lima de; RODRIGUES, Cintia Damasceno dos Santos; FONSECA, Vagner; RIBEIRO, Mario Sergio; BRANDAO, Ana Paula; CUNHA, Rivaldo Venancio da; DIAS, Ana Isabel; BOAS, Lucy Santos Vilas; FELIX, Alvina Clara; PEREIRA, Maira Alves; PINTO, Luzia Maria de Oliveira; SAKUNTABHAI, Anavaj; FILIPPIS, Ana Maria Bispo de
    Intrahost genetic diversity is thought to facilitate arbovirus adaptation to changing environments and hosts, and it might also be linked to viral pathogenesis. Dengue virus serotype 2 (DENV-2) has circulated in Brazil since 1990 and is associated with severe disease and explosive outbreaks. Intending to shed light on the viral determinants for severe dengue pathogenesis, we sought to analyze the DENV-2 intrahost genetic diversity in 68 patient cases clinically classified as dengue fever (n = 31), dengue with warning signs (n = 19), and severe dengue (n = 18). Unlike previous DENV intrahost diversity studies whose approaches employed PCR, here we performed viral whole-genome deep sequencing from clinical samples with an amplicon-free approach, representing the real intrahost diversity scenario. Striking differences were detected in the viral population structure between the three clinical categories, which appear to be driven mainly by different infection times and selection pressures, rather than being linked with the clinical outcome itself. Diversity in the NS2B gene, however, showed to be constrained, irrespective of clinical outcome and infection time. Finally, 385 non-synonymous intrahost single-nucleotide variants located along the viral polyprotein, plus variants located in the untranslated regions, were consistently identified among the samples. Of them, 124 were exclusively or highly detected among cases with warning signs and among severe cases. However, there was no variant that by itself appeared to characterize the cases of greater severity, either due to its low intrahost frequency or the conservative effect on amino acid substitution. Although further studies are necessary to determine their real effect on viral proteins, this heightens the possibility of epistatic interactions. The present analysis represents an initial effort to correlate DENV-2 genetic diversity to its pathogenic potential and thus contribute to understanding the virus's dynamics within its human host.
  • article 22 Citação(ões) na Scopus
    CD8+T Lymphocyte Expansion, Proliferation and Activation in Dengue Fever
    (2015) MATOS, Andreia Manso de; CARVALHO, Karina Inacio; ROSA, Daniela Santoro; VILLAS-BOAS, Lucy Santos; SILVA, Wanessa Cardoso da; RODRIGUES, Celia Luiza de Lima; OLIVEIRA, Olimpia Massae Nakasone Peel Furtado; LEVI, Jose Eduardo; ARAUJO, Evaldo Stanislau Affonso; PANNUTI, Claudio Sergio; LUNA, Expedito Jose Albuquerque; KALLAS, Esper George
    Dengue fever induces a robust immune response, including massive T cell activation. The level of T cell activation may, however, be associated with more severe disease. In this study, we explored the level of CD8+ T lymphocyte activation in the first six days after onset of symptoms during a DENV2 outbreak in early 2010 on the coast of Sao Paulo State, Brazil. Using flow cytometry we detected a progressive increase in the percentage of CD8+ T cells in 74 dengue fever cases. Peripheral blood mononuclear cells from 30 cases were thawed and evaluated using expanded phenotyping. The expansion of the CD8+ T cells was coupled with increased Ki67 expression. Cell activation was observed later in the course of disease, as determined by the expression of the activation markers CD38 and HLA-DR. This increased CD8+ T lymphocyte activation was observed in all memory subsets, but was more pronounced in the effector memory subset, as defined by higher CD38 expression. Our results show that most CD8+ T cell subsets are expanded during DENV2 infection and that the effector memory subset is the predominantly affected sub population.
  • article 17 Citação(ões) na Scopus
    First case report of monkeypox in Brazil: clinical manifestations and differential diagnosis with sexually transmitted infections
    (2022) LIMA, Evelyn Lepka de; BARRA, Luiz Alberto Costa; BORGES, Luciana Marques Sansao; MEDEIROS, Lucas Alberto; TOMISHIGE, Marcia Y. S.; SANTOS, Lucas de Souza Loureiro Abbud; SILVA, Anderson Jose Dias da; RODRIGUES, Camila Cristina Martini; AZEVEDO, Luiz Cesar Fernandes de; VILLAS-BOAS, Lucy Santos; SILVA, Camila Alves Maia da; COLETTI, Thais Moura; MANULI, Erika R.; CLARO, Ingra Morales; ROMANO, Camila Malta; RAMUNDO, Mariana Severo; MOUTINHO, Tomas; SABINO, Ester Cerdeira; LINDOSO, Jose Angelo Lauletta; FIGUEIREDO-MELLO, Claudia
    In 2022, an outbreak of monkeypox is being reported in non-endemic areas, with unusual clinical manifestations. The detailed clinical description of the first patient that received the diagnosis of monkeypox in Brazil is reported here, whose clinical manifestations can easily lead to misdiagnosis of sexually transmitted infections. A 41 years old male presented to an emergency room with a vesicular rash with eight days of evolution. He had traveled to Portugal and Spain and reported non-penetrative sexual involvement with three different male individuals. On the third day of symptoms, he sought medical care and received empirical treatment directed to sexually transmitted infections. As the symptoms did not improve, he sought medical attention at an infectious disease referral center presenting, on admission, an ulcerated penile lesion with central necrotic crusts, a disseminated pleomorphic skin rash and an oropharyngeal ulcer. The monkeypox diagnosis was suspected due to the characteristics of the lesions and the history of intimate contact with casual partners, and it was later confirmed by sequencing the almost complete monkeypox genome. The patient was hospitalized for pain control, which required opiate administration. He developed a secondary bacterial infection on the penile lesions, which were treated with oral antibiotics. He was discharged after 14 days, with lesions in process of re-epithelialization. Given the current outbreak, we must consider the possibility of monkeypox in patients with suggestive lesions, anywhere on the body (including the genitals), added to an epidemiological link or history of intimate contact with strangers or casual partners.
  • conferenceObject
    (2014) AMBATI, A.; TESTA, L.; LJUNGMAN, P.; BOAS, L. S. Vilas; AOUN, M.; MAEURER, M.; MACHADO, C.