ELIA TAMASO ESPIN GARCIA CALZOLARI
Projetos de Pesquisa
Unidades Organizacionais
Departamento de Patologia, Faculdade de Medicina - Docente
LIM/59 - Laboratório de Biologia Celular, Hospital das Clínicas, Faculdade de Medicina
LIM/59 - Laboratório de Biologia Celular, Hospital das Clínicas, Faculdade de Medicina
7 resultados
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- Using EM data to understand COVID-19 pathophysiology(2021) DOLHNIKOFF, Marisa; DUARTE-NETO, Amaro N.; SALDIVA, Paulo H. N.; CALDINI, Elia G.
- 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 daThe 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.
- 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, MarisaBackground: 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.
- 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, MarisaBackground: 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).
- Nicotinamide attenuates streptozotocin-induced diabetes complications and increases survival rate in rats: role of autonomic nervous system(2021) CRUZ, Paula L.; MORAES-SILVA, Ivana C.; RIBEIRO, Amanda A.; MACHI, Jacqueline F.; MELO, Marcelo Dantas Tavares de; SANTOS, Fernando dos; SILVA, Maikon Barbosa da; STRUNZ, Celia Maria Cassaro; CALDINI, Elia Garcia; IRIGOYEN, Maria-ClaudiaBackground To evaluate the effect of nicotinamide prior to streptozotocin-induced (STZ) diabetes in baroreflex sensitivity and cardiovascular autonomic modulation, and its association with hemodynamics and metabolic parameters. Methods Methods: Male Wistar rats were divided into control (Cont) and STZ-induced diabetes (Diab). Half of the rats from each group received a single dose of nicotinamide (100 mg/Kg) before STZ injection (Cont+NicA and Diab+NicA). All groups were followed-up for 5 weeks. Results Body weight loss of more than 40% was observed in Diab throughout the period (Diab: 271.00 +/- 12.74 g; Diab+NicA: 344.62 +/- 17.82). Increased glycemia was seen in Diab rats (541.28 +/- 18.68 mg/dl) while Diab+NicA group had a slight decrease (440.87 +/- 20.96 mg/dl). However, insulin resistance was observed only in Diab. In relation to Cont, heart rate, mean blood pressure and diastolic function were reduced when compared to Diab, together with parasympathetic modulation and baroreflex sensitivity. All of these parameters were improved in Diab+NicA when compared to Diab. Improved baroreflex sensitivity and parasympathetic modulation were correlated with glycemia, insulin resistance, and body weight mass. Additionally, Diab+NicA group increased survival rate. Conclusions Results suggest that the association of nicotinamide in STZ-induced diabetic rats prevents most of the expected derangements mainly by preserving parasympathetic and baroreflex parameters.
- Pre-coating decellularized liver with HepG2-conditioned medium improves hepatic recellularization(2021) CAIRES-JUNIOR, Luiz Carlos; GOULART, Ernesto; TELLES-SILVA, Kayque Alves; ARAUJO, Bruno Henrique Silva; MUSSO, Camila Manso; KOBAYASHI, Gerson; OLIVEIRA, Danyllo; ASSONI, Amanda; CARVALHO, Valdemir Melechco; RIBEIRO-JR, Antonio Fernando; ISHIBA, Renata; BRAGA, Karina Andrighetti Oliveira; NEPOMUCENO, Natalia; CALDINI, Elia; RANGEL, Thadeu; RAIA, Silvano; I, Peter Lelkes; ZATZ, MayanaLiver transplantation from compatible donors has been the main therapy available for patients with irreversible hepatic injuries. Due to the increasing shortage of organs suitable for transplantation, tissue engineering technologies are important alternatives or surrogate approaches for the future of human organ transplantations. New bioengineering tools have been designed to produce decellularized organs (i.e. scaffolds) which could be recellularized with human cells. Specifically, there is an unmet need for developing reproducible protocols for inducing better cellular spreading in decellularized liver scaffolds. The aim of the present work was to investigate the possibility to improve liver scaffold recellularization by pre-coating decellularized tissue scaffolds with HepG2-conditioned medium (CM). Furthermore, we evaluated the capability of commercial human liver cells (HepG2) to adhere to several types of extracellular matrices (ECM) as well as CM components. Wistar rat livers were decellularized and analyzed by histology, scanning electron microscopy (SEM), immunohistochemistry and residual DNA-content analysis. Human induced pluripotent stem cells (hiPSCs)-derived mesenchymal cells (hiMSCs), and human commercial hepatic (HepG2) and endothelial (HAEC) cells were used for liver scaffold recellularization with or without CM pre-coating. Recellularization occurred for up to 5 weeks. Hepatic tissues and CM were analyzed by proteomic assays. We show that integrity and anatomical organization of the hepatic ECM were maintained after decellularization, and proteomic analysis suggested that pre-coating with CM enriched the decellularized liver ECM. Pre-coating with HepG2-CM highly improved liver recellularization and revealed the positive effects of liver ECM and CM components association.
- Skin Changes Due to Massive Weight Loss: Histological Changes and the Causes of the Limited Results of Contouring Surgeries(2021) I, Rodrigo Rocha; CINTRA JUNIOR, Wilson; MODOLIN, Miguel L. A.; TAKAHASHI, Giulia G.; CALDINI, Elia T. E. G.; GEMPERLI, RolfPurpose Outcomes of body contouring surgeries in patients who previously had obesity are limited because of the loss of skin quality. This study aimed to evaluate the morphometric characteristics of collagen and elastic fibers of the skin in the abdominal epigastric region of patients who had massive weight loss following bariatric surgery and compared such with the skin characteristics of patients with morbid obesity. Methods This observational study compared skin fragments from the epigastric region of 20 patients who had massive weight loss due to bariatric surgery and 20 patients with morbid obesity. The morphometric analysis was performed on the collagen system using the Picrosirius/polarized light method and on the elastic system using the Weigert's resorcin-fuchsin method. Results Reduction of thick collagen fibers (p = 0.048), increased thin collagen fibers (p = 0.0085), and increased elastic fiber density (p < 0.001) were observed in the massive weight loss group. No differences were found between the groups regarding mean age (p = 0.917) and total amount of collagen fibers (p = 0.3619). Structural dermis alterations in the massive weight loss group demonstrated collagenous remodeling, with consequent reduction of thick, organized, structured, and directed fibers in favor of thin, misaligned, and loosely arranged fibers. Weight loss was also associated with increased skin elasticity. Conclusion The morphometric changes in the collagen and elastic system scientifically explained the already established clinical perception of cutaneous alterations in patients who had massive weight loss following bariatric surgeries.