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- ItemPatogênese molecular da COVID-19: o envolvimento de fatores genéticos, hemostáticos e imunológicos na evolução de quadros graves, desenvolvimento de coagulopatias e COVID Longa.(Universidade Federal do Espírito Santo, 2025-03-21) Silva, Danielle Ribeiro Campos da Silva; Meira, Débora Dummer; https://orcid.org/0000-0002-6092-2459; http://lattes.cnpq.br/7199119599752978; Louro, Iúri Drumond; https://orcid.org/0000-0001-5160-9615; http://lattes.cnpq.br/3817361438227180; https://orcid.org/0009-0004-3821-7376; http://lattes.cnpq.br/8642525842646026; Paula, Flávia de; https://orcid.org/0000-0001-8679-2982; http://lattes.cnpq.br/7913201450663683; Figueiras, Paulo Roberto; https://orcid.org/0000-0002-6068-1982; ; Prezotti, Alessandra Nunes Loureiro; https://orcid.org/0000-0003-1079-7140; http://lattes.cnpq.br/9069306254016368; Pereira, Fausto Edmundo Lima; https://orcid.org/0000-0002-9219-3537; http://lattes.cnpq.br/4065537941002091Introduction: The development of the inflammatory process triggered by the SARS-CoV-2 virus leads to abnormal activation of the coagulation system, pathologically manifesting as generalized endothelial dysfunction in small vessels and microthrombosis in the alveolar region. Following the pandemic, persistent symptoms and sequelae are frequently associated with this infection—referred to as “Long COVID”—and generally have a negative impact on individuals’ daily activities. The investigation of coagulation-related markers, cytokines, antiphospholipid antibodies, and genotyping of interleukins and thrombophilia-associated genes that influence hyperinflammation, endothelial dysfunction, and hypoxia is highly relevant for the prognosis of surviving patients. Objective: To investigate molecular and genetic alterations in the hemostatic and inflammatory systems, ABO blood group, and the presence of acquired antibodies in patients who tested positive for COVID-19, as well as the association of these factors with disease severity, potential occurrence of thrombotic complications, and the development of Long COVID. Methods: A total of 226 unvaccinated volunteers (considering two doses) with RT-PCR-confirmed COVID-19 were recruited between November 2020 and December 2021. The cohort was divided into three groups: Mild (60), Moderate (65), and Severe (101), according to WHO guidelines. All participants signed an Informed Consent Form, completed a questionnaire, and had blood samples collected for analysis. The questionnaire assessed relevant preexisting clinical conditions, medical record data, symptoms, and the occurrence of sequelae. Laboratory tests were conducted to assess thrombosis, blood typing, hemostatic/inflammatory biomarkers, and genotyping. Participants whose test results were outside reference ranges were re-evaluated six months later, and after two years, they completed a follow-up questionnaire focused on Long COVID and quality of life. Appropriate statistical tests were used for data analysis. Results: A significant relationship was identified between disease severity and sociodemographic and biomarker data. Statistically significant factors included: age (P age (P<0.0001), lower educational level/ social vulnerability (P<0.0001), ethnicity (P=0.003), overweight (P<0.0001), MTHFR rs1801133 polymorphism (P=0.035), previous cardiovascular disease (P=0.002), diabetes mellitus (DM) (P=0.006), Factor VIII (FVIII) (P=0.046), von Willebrand Factor (vWF) (P=0.0020), and D-dimer (DD) (P<0.0001). After six months, FVIII (P=0.0005), vWF (P=0.0022), and DD (P<0.0001) levels significantly decreased, but only DD returned to normal levels.Sequelae affecting the musculoskeletal, cardiorespiratory, and vascular systems were investigated. Individuals with chronic pulmonary disease (CPD) and DM had higher chances of developing joint pain and myalgia. CPD was also associated with dyspnea and palpitations. Elevated DD levels contributed to palpitations. Prolonged dry cough was associated with lower education and non-white ethnicity. Women were more likely to develop phlebitis. FVIII was associated with a higher risk of cramps, and regular physical activity showed a protective effect against swelling. The EQ-5D-5L questionnaire assessed current quality of life, with the most severely affected participants reporting greater impairment across all measured domains. Conclusion: The findings suggest that endothelial dysfunction is a central factor and may contribute to the development of Long COVID.
- ItemAnálise de custo do tratamento do câncer da cavidade oral e orofaringe: Uso do método de microcusteio por bottom-up para dados de um hospital de referência no Brasil(Universidade Federal do Espírito Santo, 2024-03-11) Zorzaneli, Willene dos Santos Machado; Zara, Ana Laura Amâncio de Sene; Zeidler, Sandra Ventorin von; https://orcid.org/0000-0002-8897-5747; http://lattes.cnpq.br/5785612863130498; Paula, Flavia de; Guimarães, Michele Ferreira; Silva, Everton Nunes daCost analysis in OC/OPC is scarce, however, necessary since cost can be an essential tool for decision-making. This study aimed to estimate the direct costs of oral and oropharyngeal squamous cell carcinoma (OC/OPC) in a state in southeastern Brazil. To this end, a cost of illness study was carried out based on the bottom-up microcosting method as a partial economic assessment, considering the direct medical costs related to outpatients and inpatients from the perspective of reimbursement rates from the Unified Health System. Data were collected from the medical records of 41 randomized patients recruited, between 2013 and 2018. All cases were confirmed as squamous cell carcinoma of the mouth and oropharynx (C02 to C06, C09 and C10 according to the International Classification of Diseases for Oncology, 3rd Edition). Each cost value was adjusted for annual accumulated inflation. The statistical analysis of descriptive analyses, parametric tests, ANOVA and Student t test and non-parametric tests, Kruskal Wallis and Mann Whitney, was used using the SPSS® Statistical program. To adjust the differences in confidence intervals at 95%, the bootstrap method was used for 5.000 resamples using the Stata® statistical program. For all statistical tests, the significance level was set at 5%. Healthcare system cost estimates for 41 patients with OC/OPC amounted to a total cost of I$164,671.07 with an average of IS$4,042.5 per patient. 22,814 procedures and uses of health services were carried out. The sample was mostly made up of men (>80%) and aged between 51 and 60 years (41.5%). Carcinomas of the base of the tongue and floor of the mouth had the highest costs, I$4,288.25 and 6,586.88, respectively. Clinical stage III was the one with the most procedures and highest costs, I$5,415.11 (p<0.001) per patient. From the perspective of the Unified Health System, there is a high-cost demand for OC/OPC treatment, especially in advanced stages of the disease. This study contributes to decision- making regarding treatment as well as the implementation of programs aimed at early detection of diseases
- ItemCâncer de cabeça e pescoço : tendência de mortalidade e análise comparativa das técnicas de hibridização in situ, RT-qPCR e p16 para diagnóstico de infecção por HPV(Universidade Federal do Espírito Santo, 2024-12-17) Sant'Anna, Jéssica Graça; Zeidler, Sandra Lúcia Ventorin von ; https://orcid.org/0000-0002-8897-5747; http://lattes.cnpq.br/5785612863130498; https://orcid.org/0000-0001-9759-2978; http://lattes.cnpq.br/2132178774575269; Paula, Flavia de ; https://orcid.org/0000-0001-8679-2982; http://lattes.cnpq.br/7913201450663683; Curado, Maria Paula ; https://orcid.org/0000-0001-8172-2483; http://lattes.cnpq.br/3397823736381748; Silva, Melissa de Freitas Cordeiro ; https://orcid.org/0000-0002-7709-5912; http://lattes.cnpq.br/6753487805311437; Podesta, Olívia Perim Galvão de; https://orcid.org/0000-0001-6669-5915; http://lattes.cnpq.br/2110363960968286In Brazil, regional disparities in mortality rates from head and neck squamous cell carcinoma (HNSCC) are high. Infection with high-risk oncogenic human papillomavirus (HR-HPV) is an important risk factor, associated with an increase in cases of oropharyngeal squamous cell carcinoma (OPSCC) in Brazil and globally. This study analyzed mortality trends from HNSCC between 1998 and 2017 in Espírito Santo, Brazil (ES-BR), and methods for detecting HR-HPV in OPSCC using messenger RNA (mRNA) E6 by RT-qPCR, mRNA E6/E7 by in situ hybridization (RNA-ISH), and immunohistochemistry for p16 expression (IHC-p16) in Espírito Santo, Brazil (ES-BR) and Bogotá, Colombia (BO-CO). The mortality trend methodology was conducted in reference hospitals in ES-BR, using mortality data from DATASUS, the Mortality Information System (SIM), and data from the Brazilian Institute of Geography and Statistics (IBGE). Clinicopathological characteristics and HPV status of the studied sites were analyzed, and HPV detection methods were compared using RNA-ISH and IHC-p16 with RT-qPCR as the standard for sensitivity and specificity analysis. Between 1998 and 2017, mortality from HNSCC increased in ES, especially among men, rising from 48.96 to 58.24 deaths per 100,000 inhabitants. Regarding anatomical subsites, men showed an increase in deaths from oral cavity, oropharyngeal, and laryngeal cancers, while women’s increase was limited to oral cavity cancer. Regarding HR-HPV, samples from ES showed lower frequencies of infection compared to those from BO. Lymph node metastasis was associated with a worse prognosis, while HPV-positive patients had better survival. Smoking negatively impacted survival. Detection methods for HR-HPV, such as IHC p16, RNA-ISH, and RT-qPCR, showed statistical significance. The results show an increase in HNSCC mortality in ES, especially among men and in OPSCC. The IHC p16 technique had high sensitivity, while RNA-ISH showed high specificity. Regional strategies and advanced diagnostics are crucial for managing HNSCC
- ItemRegulación de plantas genéticamente editadas y selección del gen SAMT para la edición de carica papaya buscando resistencia a meleira(Universidade Federal do Espírito Santo, 2024-05-09) Gaitán, Julia Bibiana Merchán; Rodrigues, Silas Pessini; Fernandes, Patrícia Machado Bueno; https://orcid.org/0000-0003-2695-3638; Ventura, José Aires; Tarazi, Roberto; Pérez Brito, Daisy De La Caridad; Rodrigues, Juliany Cola FernandesThis paper addresses the regulation of one of the most modern technologies for the control of plant pathologies, the CRISPR (Clustered regularly interspaced short palindromic repeats) technique, which is based on the action of a Cas protein and an gRNA that together can target specific areas of the DNA to modify them, through a systematic review of the regulatory systems of different countries. The different approaches adopted by countries in the Latin American region to regulate CRISPR technology were analyzed, taking into account the opportunities it offers and the possible associated ethical and social risks. So far, Brazil has analyzed the case-by case evaluation process of genetically modified crops by the regulatory committee CTNBio for the use of new plant breeding technologies to determine whether it would be considered a GMO or conventional breeding. This analysis was carried out because there is great interest in using CRISPR technology to develop new plant cultivars, especially plants relevant to Brazilian fruit growing. The Carica papaya cultivar, is of particular interest because of limitations in its production, due to the plant's susceptibility to the virus complex that causes papaya meleira disease, is economically and biologically relevant, as viruses have been identified in plant latex, a fluid derived from laticifers, which occur in more than 20 plant families and on which there is a body of information that points to laticifers as plant response systems to viruses. For this reason, a detailed review was conducted on the role of laticiferans of different species, examining the ability of both pro- and antiviral latex to protect plants, as well as its potential to be used as a natural source of defense molecules against insect vectors and fungal infections. The potential use of CRISPR in response to control viruses affecting laticifers was also evaluated through a compilation of articles targeting gene editing with specific expression or related to laticifer biology, confirming that it is possible to modulate laticifer function using CRISPR. The relationship of the PMeV complex in the C. papaya L. plant was discussed and the importance of the cloning of the Salicylate O-methyltransferase gene in the defense against this viral complex are discussed, due to its presence in the plant defense mechanisms related to laticifers. Describing experimental advances in the development of a protocol to silence the salicylate O-methyltransferase gene of C. papaya by means of the Golden Gate Assembly, detailing the design of gRNA for the construction and multiplication of gene editing vectors, intermediate steps in the process of gene silencing of the SAMT gene for the control of meleira disease in papaya, resulting in optimal conditions for the use of CRISPR technology in C. papaya plants.
- ItemDesenvolvimento de biomaterial ósseo descelularizado enriquecido com hidrogel de osso fetal bovino(Universidade Federal do Espírito Santo, 2024-10-10) Barros, Rodolpho José da Silva; Nogueira, Breno Valentim ; https://orcid.org/0000-0002-2199-0635; http://lattes.cnpq.br/0011229320439147; https://orcid.org/0000-0001-6214-733X; http://lattes.cnpq.br/5652361736715531; Maranduba, Carlos Magno da Costa ; https://orcid.org/0000-0001-7327-1934; http://lattes.cnpq.br/4763153859701731; Oliveira, Jairo Pinto de ; https://orcid.org/0000-0001-7595-1183; http://lattes.cnpq.br/2228283301316218; Guimarães, Marco Cesar Cunegundes ; https://orcid.org/0000-0003-2146-0180; http://lattes.cnpq.br/0261991057482057; Aires, Rafaela; https://orcid.org/0000-0002-9532-9401; http://lattes.cnpq.br/5449202639864996The global bone graft material market is projected to grow at a rate of 6-7% annually until 2032. Conventional bone grafting strategies are no longer considered sufficient or adequate for most treatments. The development of decellularized biomaterials derived from animal tissues (xenografts) offers a promising and viable strategy. The structural similarity between animal and human bones, along with the decellularization technique, can result in a high-quality biomaterial that provides mechanical support, preserves the organic matrix, and supplies essential biological stimuli for tissue regeneration. Adult and fetal bovine bones were decellularized to produce biomaterials (blocks, granules, and hydrogel) with residual DNA levels lower than 50 ng/mg and SDS levels below 2 μg/mg of dry tissue. Thermogravimetric assays (DSC and TGA) and infrared spectroscopy (FTIR and Raman) demonstrated the adequate preservation of both the mineral and organic bone matrices. Scanning electron microscopy (SEM) and elemental composition analysis (EDS and CHNS-O) confirmed the preservation of the microstructure and biochemical composition of the bone matrix. pH stability, absence of heavy metals, and sterility were verified. Comparative tests showed that the decellularized biomaterials, particularly those derived from fetal bone, exhibit superior porosity, density, mechanical support capacity, and organic composition compared to other biomaterials. In vitro tests revealed no cytotoxicity and a higher ability to stimulate the proliferation of mesenchymal cells and fibroblasts. The results indicate that the production of decellularized bovine bone biomaterials coated with fetal hydrogel could contribute to better biocompatibility of implants and help meet the global demand for bone graft materials