ELIA TAMASO ESPIN GARCIA CALZOLARI

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Departamento de Patologia, Faculdade de Medicina - Docente
LIM/59 - Laboratório de Biologia Celular, Hospital das Clínicas, Faculdade de Medicina

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  • conferenceObject
    ELEVATED ROS LEVELS AND DNA FRAGMENTATION IN SPERM FROM CONVALESCENT MEN IN SARS-COV-2 INFECTION
    (2022) HALLAK, Jorge; BERNARDES, Felipe; TEIXEIRA, Thiago Afonso Carvalho Celestino; SALDIVA, Paulo Hilario Nascimento; KALLAS, Esper Georges; CALDINI, Elia Tamaso Espin Garcia; DUARTE NETO, Amaro Nunes; FAQUINETI, Heloisa; JESUS, Vinicius Luiz Menezes; GUTIERREZ, Raul Segundo Sanchez; DREVET, Joel R.
  • article 2 Citação(ões) na Scopus
    Using EM data to understand COVID-19 pathophysiology
    (2021) DOLHNIKOFF, Marisa; DUARTE-NETO, Amaro N.; SALDIVA, Paulo H. N.; CALDINI, Elia G.
  • article 46 Citação(ões) na Scopus
    Testicular pathology in fatal COVID-19: A descriptive autopsy study
    (2022) DUARTE-NETO, Amaro N.; TEIXEIRA, Thiago A.; CALDINI, Elia G.; KANAMURA, Cristina T.; GOMES-GOUVEA, Michele S.; SANTOS, Angela B. G. dos; MONTEIRO, Renata A. A.; PINHO, Joao R. R.; MAUAD, Thais; SILVA, Luiz F. F. da; SALDIVA, Paulo H. N.; DOLHNIKOFF, Marisa; LEITE, Katia R. M.; HALLAK, Jorge
    Background Multi-organ damage is a common feature of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, going beyond the initially observed severe pneumonia. Evidence that the testis is also compromised is growing. Objective To describe the pathological findings in testes from fatal cases of COVID-19, including the detection of viral particles and antigens, and inflammatory cell subsets. Materials and methods Postmortem testicular samples were obtained by percutaneous puncture from 11 deceased men and examined by reverse-transcription polymerase chain reaction (RT-PCR) for RNA detection and by light and electron microscopy (EM) for SARS-CoV-2. Immunohistochemistry (IHC) for the SARS-CoV-2 N-protein and lymphocytic and histiocytic markers was also performed. Results Eight patients had mild interstitial orchitis, composed mainly of CD68+ and TCD8+ cells. Fibrin thrombi were detected in five cases. All cases presented congestion, interstitial edema, thickening of the tubular basal membrane, decreased Leydig and Sertoli cells with reduced spermatogenesis, and strong expression of vascular cell adhesion molecule (VCAM) in vessels. IHC detected SARS-Cov-2 antigen in Leydig cells, Sertoli cells, spermatogonia, and fibroblasts in all cases. EM detected viral particles in the cytoplasm of fibroblasts, endothelium, Sertoli and Leydig cells, spermatids, and epithelial cells of the rete testis in four cases, while RT-PCR detected SARS-CoV-2 RNA in three cases. Discussion and conclusion The COVID-19-associated testicular lesion revealed a combination of orchitis, vascular changes, basal membrane thickening, Leydig and Sertoli cell scarcity, and reduced spermatogenesis associated with SARS-CoV-2 local infection that may impair hormonal function and fertility in men.
  • article 59 Citação(ões) na Scopus
    Salivary glands are a target for SARS-CoV-2: a source for saliva contamination
    (2021) MATUCK, Bruno Fernandes; DOLHNIKOFF, Marisa; DUARTE-NETO, Amaro Nunes; MAIA, Gilvan; GOMES, Sara Costa; SENDYK, Daniel Isaac; ZARPELLON, Amanda; ANDRADE, Nathalia Paiva de; MONTEIRO, Renata Aparecida; PINHO, Joao Renato Rebello; GOMES-GOUVEA, Michele Soares; SOUZA, Suzana C. O. M.; KANAMURA, Cristina; MAUAD, Thais; SALDIVA, Paulo Hilario Nascimento; BRAZ-SILVA, Paulo H.; CALDINI, Elia Garcia; SILVA, Luiz Fernando Ferraz da
    The ability of the new coronavirus SARS-CoV-2 to spread and contaminate is one of the determinants of the COVID-19 pandemic status. SARS-CoV-2 has been detected in saliva consistently, with similar sensitivity to that observed in nasopharyngeal swabs. We conducted ultrasound-guided postmortem biopsies in COVID-19 fatal cases. Samples of salivary glands (SGs; parotid, submandibular, and minor) were obtained. We analyzed samples using RT-qPCR, immunohistochemistry, electron microscopy, and histopathological analysis to identify SARS-CoV-2 and elucidate qualitative and quantitative viral profiles in salivary glands. The study included 13 female and 11 male patients, with a mean age of 53.12 years (range 8-83 years). RT-qPCR for SARS-CoV-2 was positive in 30 SG samples from 18 patients (60% of total SG samples and 75% of all cases). Ultrastructural analyses showed spherical 70-100 nm viral particles, consistent in size and shape with the Coronaviridae family, in the ductal lining cell cytoplasm, acinar cells, and ductal lumen of SGs. There was also degeneration of organelles in infected cells and the presence of a cluster of nucleocapsids, which suggests viral replication in SG cells. Qualitative histopathological analysis showed morphologic alterations in the duct lining epithelium characterized by cytoplasmic and nuclear vacuolization, as well as nuclear pleomorphism. Acinar cells showed degenerative changes of the zymogen granules and enlarged nuclei. Ductal epithelium and serous acinar cells showed intense expression of ACE2 and TMPRSS receptors. An anti-SARS-CoV-2 antibody was positive in 8 (53%) of the 15 tested cases in duct lining epithelial cells and acinar cells of major SGs. Only two minor salivary glands were positive for SARS-CoV-2 by immunohistochemistry. Salivary glands are a reservoir for SARS-CoV-2 and provide a pathophysiological background for studies that indicate the use of saliva as a diagnostic method for COVID-19 and highlight this biological fluid's role in spreading the disease. (C) 2021 The Pathological Society of Great Britain and Ireland.
  • article 0 Citação(ões) na Scopus
    Ultrastructural characterization of alveolar microvascular damage in severe COVID-19 respiratory failure
    (2023) NEGRI, Elnara Marcia; BENCHIMOL, Marlene; MAUAD, Thais; DUARTE-NETO, Amaro Nunes; GOTTARDI, Maiara; SILVA, Luiz Fernando Ferraz da; SALDIVA, Paulo Hilario Nascimento; DOLHNIKOFF, Marisa; SOUZA, Wanderley de; CALDINI, Elia Garcia
    Endothelial dysfunction is a key phenomenon in COVID-19, induced by direct viral endothelial infection and secondary inflammation, mainly affecting the microvascular circulation. However, few studies described the subcellular aspects of the lung microvasculature and the associated thrombotic phenomena, which are widely present in severe COVID-19 cases. To that end, in this transversal observational study we performed transmission and scanning electron microscopy in nine lung samples of patients who died due to COVID-19, obtained via minimally invasive autopsies in Sao Paulo, Brazil, in 2020. All patients died due to acute respiratory failure and had microvascular thrombosis at histology. Electron microscopy revealed areas of endothelial damage with basal lamina disruption and virus infection in endothelial cells. In the capillary lumens, the ultrastructure of the thrombi is depicted, with red blood cells stacking, dysmorphism and hemolysis, fibrin meshworks, and extracellular traps. Our description illustrates the complex pathophysiology of microvascular thrombosis at the cellular level, which leads to some of the peculiar characteristics of severe COVID-19.
  • article 5 Citação(ões) na Scopus
    Ultrasound-Guided Minimally Invasive Tissue Sampling: A Minimally Invasive Autopsy Strategy During the COVID-19 Pandemic in Brazil, 2020
    (2021) DUARTE-NETO, Amaro Nunes; SILVA, Luiz Fernando Ferraz da; MONTEIRO, Renata Aparecida de Almeida; FILHO, Jair Theodoro; LEITE, Thabata Larissa Luciano Ferreira; MOURA, Catia Sales de; GOMES-GOUVEA, Michele Soares; PINHO, Joao Renato Rebellho; KANAMURA, Cristina Takami; OLIVERIA, Ellen Pierre de; BISPO, Kely Cristina Soares; ARRUDA, Cassia; SANTOS, Aline Brito dos; AQUINO, Flavia Cristina Goncalves; CALDINI, Elia Garcia; MAUAD, Thais; SALDIVA, Paulo Hilario Nascimento; DOLHNIKOFF, Marisa
    Background: Minimally invasive autopsies, also known as minimally invasive tissue sampling (MITS), have proven to be an alternative to complete diagnostic autopsies (CDAs) in places or situations where this procedure cannot be performed. During the coronavirus disease 2019 (COVID-19) pandemic, CDAs were suspended by March 2020 in Brazil to reduce biohazard. To contribute to the understanding of COVID-19 pathology, we have conducted ultrasound (US)-guided MITS as a strategy. Methods: This case series study includes 80 autopsies performed in patients with COVID-19 confirmed by laboratorial tests. Different organs were sampled using a standardized MITS protocol. Tissues were submitted to histopathological analysis as well as immunohistochemical and molecular analysis and electron microscopy in selected cases. Results: US-guided MITS proved to be a safe and highly accurate procedure; none of the personnel were infected, and accuracy ranged from 69.1% for kidney, up to 90.1% for lungs, and reaching 98.7% and 97.5% for liver and heart, respectively. US-guided MITS provided a systemic view of the disease, describing the most common pathological findings and identifying viral and other infectious agents using ancillary techniques, and also allowed COVID-19 diagnosis confirmation in 5% of the cases that were negative in premortem and postmortem nasopharyngeal/oropharyngeal swab real-time reverse-transcription polymerase chain reaction. Conclusions: Our data showed that US-guided MITS has the capacity similar to CDA not only to identify but also to characterize emergent diseases.
  • article 87 Citação(ões) na Scopus
    An autopsy study of the spectrum of severe COVID-19 in children: From SARS to different phenotypes of MIS-C
    (2021) DUARTE-NETO, Amaro Nunes; CALDINI, Elia Garcia; GOMES-GOUVEA, Michele Soares; KANAMURA, Cristina Takami; MONTEIRO, Renata Aparecida de Almeida; FERRANTI, Juliana Ferreira; VENTURA, Andrea Maria Cordeiro; REGALIO, Fabiane Aliotti; FIORENZANO, Daniela Matos; GIBELLI, Maria Augusta Bento Cicaroni; CARVALHO, Werther Brunow de; LEAL, Gabriela Nunes; PINHO, Joao Renato Rebello; DELGADO, Artur Figueiredo; CARNEIRO-SAMPAIO, Magda; MAUAD, Thais; SILVA, Luiz Fernando Ferraz da; SALDIVA, Paulo Hilario Nascimento; DOLHNIKOFF, Marisa
    Background: COVID-19 in children is usually mild or asymptomatic, but severe and fatal paediatric cases have been described. The pathology of COVID-19 in children is not known; the proposed pathogenesis for severe cases includes immune-mediated mechanisms or the direct effect of SARS-CoV-2 on tissues. We describe the autopsy findings in five cases of paediatric COVID-19 and provide mechanistic insight into the mechanisms involved in the pathogenesis of the disease. Methods: Children and adolescents who died with COVID-19 between March 18 and August 15, 2020 were autopsied with a minimally invasive method. Tissue samples from all vital organs were analysed by histology, electron microscopy (EM), reverse-transcription polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC). Findings: Five patients were included, one male and four female, aged 7 months to 15 years. Two patients had severe diseases before SARS-CoV-2 infection: adrenal carcinoma and Edwards syndrome. Three patients were previously healthy and had multisystem inflammatory syndrome in children (MIS-C) with distinct clinical presentations: myocarditis, colitis, and acute encephalopathy with status epilepticus. Autopsy findings varied amongst patients and included mild to severe COVID-19 pneumonia, pulmonary microthrombosis, cerebral oedema with reactive gliosis, myocarditis, intestinal inflammation, and haemophagocytosis. SARSCoV-2 was detected in all patients in lungs, heart and kidneys by at least one method (RT-PCR, IHC or EM), and in endothelial cells from heart and brain in two patients with MIS-C (IHC). In addition, we show for the first time the presence of SARS-CoV-2 in the brain tissue of a child with MIS-C with acute encephalopathy, and in the intestinal tissue of a child with acute colitis. Interpretation: SARS-CoV-2 can infect several cell and tissue types in paediatric patients, and the target organ for the clinical manifestation varies amongst individuals. Two major patterns of severe COVID-19 were observed: a primarily pulmonary disease, with severe acute respiratory disease and diffuse alveolar damage, or a multisystem inflammatory syndrome with the involvement of several organs. The presence of SARS-CoV-2 in several organs, associated with cellular ultrastructural changes, reinforces the hypothesis that a direct effect of SARS-CoV-2 on tissues is involved in the pathogenesis of MIS-C. (c) 2021 The Author(s).
  • article 186 Citação(ões) na Scopus
    SARS-CoV-2 in cardiac tissue of a child with COVID-19-related multisystem inflammatory syndrome
    (2020) DOLHNIKOFF, Marisa; FERRANTI, Juliana Ferreira; MONTEIRO, Renata Aparecida de Almeida; DUARTE-NETO, Amaro Nunes; GOMES-GOUVEA, Michele Soares; DEGASPARE, Natalia Viu; DELGADO, Artur Figueiredo; FIORITA, Carolina Montanari; LEAL, Gabriela Nunes; RODRIGUES, Regina Maria; CHAIM, Khallil Taverna; PINHO, Joao Renato Rebello; CARNEIRO-SAMPAIO, Magda; MAUAD, Thais; SILVA, Luiz Fernando Ferraz da; CARVALHO, Werther Brunow de; SALDIVA, Paulo Hilario Nascimento; CALDINI, Elia Garcia
  • conferenceObject
    INTRACELLULAR SARS-COV-2 FOUNDIN THE SPERMATOZOA OF CONVALESCENT MEN WITH NEGATIVE PCR SEMEN SAMPLES REVEALED BY ELECTRON MICROSCOPY
    (2022) HALLAK, Jorge; CALDINI, Elia Tamaso Espin Garcia; CORREA, Maria Cassia Jacintho Mendes; DOLHNIKOFF, Marisa; SALDIVA, Paulo Hilario Nascimento; KALLAS, Esper Georges; DUARTE NETO, Amaro Nunes; GUTIERREZ, Raul Segundo Sanchez; DREVET, Joel R.; TEIXEIRA, Thiago Afonso Carvalho Celestino; BERNARDES, Felipe Saraiva; FAQUINETI, Heloisa