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Agradecimentos: The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by FAEPEX/UNICAMP (Fundo de Apoio ao Ensino, Pesquisa e Extens~ao), Auxilios Emergenciais COVID-19 (grant number 2418/20) Abstract: In this cross-sectional study, we investigate the presence of Severe Acute Respiratory Syndrome Coronavirus 2 Ribonucleic Acid (SARS-CoV-2 RNA) in the tears of hospitalized COVID-19 patients. After laboratory confirmation of SARS-CoV-2 infection by reverse transcription polymerase chain reaction (RT-PCR) analysis, tear samples from both eyes of each patient were collected using conjunctival swab for RT-PCR. Detailed demographic profile, systemic and ocular symptoms, comorbidities, clinical, ancillary, and ocular manifestations were evaluated. Of the 83 patients enrolled in the study, 7 (8.43%) had SARS-CoV-2 RNA detected in the tear samples. Neutrophils’ count, C-reactive protein, and D-dimer were higher in patients with SARS-CoV-2 detected in tears than in patients without virus in ocular surface samples. One patient with SARS-CoV-2 in tears showed mild ocular eyelid edema, hyperemia, and chemosis. No relevant ocular manifestations were detected in the other patients. Although the levels of viral RNA on ocular surface samples were low for most patients (5/7), with positivity only for gene N and CT higher than 30, two patients were positive for all viral targets tested (N, E, and RpRd), with viral load near 1 105 ePFU/mL, indicating that the ocular transmission of SARS-CoV-2 is a possibility that needs to be considered, especially in the hospital environment. Further studies need to be conducted to demonstrate whether infective viral particles could be isolated from tears Aberto

Brazil currently has one of the fastest-growing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) epidemics in the world. Because of limited available data, assessments of the impact of nonpharmaceutical interventions (NPIs) on this virus spread remain challenging. Using a mobility-driven transmission model, we show that NPIs reduced the reproduction number from >3 to 1 to 1.6 in São Paulo and Rio de Janeiro. Sequencing of 427 new genomes and analysis of a geographically representative genomic dataset identified >100 international virus introductions in Brazil. We estimate that most (76%) of the Brazilian strains fell in three clades that were introduced from Europe between 22 February and 11 March 2020. During the early epidemic phase, we found that SARS-CoV-2 spread mostly locally and within state borders. After this period, despite sharp decreases in air travel, we estimated multiple exportations from large urban centers that coincided with a 25% increase in average traveled distances in national flights. This study sheds new light on the epidemic transmission and evolutionary trajectories of SARS-CoV-2 lineages in Brazil and provides evidence that current interventions remain insufficient to keep virus transmission under control in this country.

Serum samples from 150 NS1-negative (Platelia ELISA) patients presumptively diagnosed with dengue were analyzed by the TaqMan probed real-time reverse transcription PCR (TaqMan qRT-PCR) method. The qRT-PCR positive samples were tested for serotype by semi-nested RT-PCR and a qualitative immunochromatographic assay for IgG and IgM. Molecular detection methods showed 33 (22%) positive samples out of 150 NS1-antigen negative samples. Of these, 72% were collected up to day 2 after the onset of symptoms, when diagnostic sensitivity of NS1-antigen test assays is significantly enhanced. Most of the cases were not characterized as secondary infection. Twenty-eight samples were successfully serotyped, 75% of which for DENV-4, 14% for DENV-2, 7% for DENV-3 and 4% for DENV-1. These findings reaffirm the hyperendemic situation of the state of Roraima and suggest a lower sensitivity of the NS1 test, mainly when DENV-4 is the predominant serotype. Health care providers should therefore be aware of samples tested negative by NS1 antigen assays, especially when clinical symptoms and other laboratory data results show evidence of dengue infection. Amostras séricas de 150 pacientes, com diagnóstico presuntivo de dengue e resultado negativo para dengue por ELISA-NS1-Antígeno do kit Platelia™ (NS1-Ag), foram analisadas pela técnica de TaqMan Transcrição Reversa seguida da Reação em Cadeia da Polimerase em Tempo Real (qRT-PCR). As amostras positivas por qRT-PCR, foram submetidas a identificação dos sorotipos por RT-Hemi nested-PCR e a ensaio imunocromatográfico para detecção qualitativa dos anticorpos IgG e IgM. A técnica molecular apresentou como resultado 33 (22%) amostras positivas entre as 150 negativas pela detecção do NS1-Ag, destas o 72% foram coletadas até o segundo dia de início dos sintomas da doença, período de maior sensibilidade para pesquisas de NS1-Ag. A maioria dos casos não evidenciou infecção secundária. Dessas amostras, 28 foram satisfatoriamente sorotipadas sendo 75% de DENV-4, 14% de DENV-2, 7% de DENV-3 e 4% de DENV-1. Os resultados reafirmam a situação hiperendêmica do Estado de Roraima e sugerem baixa sensibilidade do NS1 test, especialmente quando o sorotipo predominante é DENV-4. Sugerimos assim, que a comunidade médica deve ser alertada no sentido de ser cautelosa com resultados de NS1-Ag negativo, principalmente quando sintomas clínicos e outros resultados laboratoriais sejam indicativos de provável infecção por dengue.

Background: The identification of groups at high risk is fundamental to determine preventive strategies for skin cancer. Destructive reactive oxygen species produced by UVA or chemical carcinogens are metabolized by a series of enzymes. Polymorphisms of genes encoding for these enzymes may produce defective proteins with a diminished ability to detoxify a wide range of carcinogens. Aims: To ascertain the influence and potential interactions of several polymorphisms of genes encoding four important antioxidant GST enzymes in the susceptibility to cancer among Brazilians. Material and methods: We compared the genotypes of Glutathione S-Transferase mu, theta, pi and omega (GSTM1, GSTT1, GSTP1 and GSTO2) in a group of 102 patients with skin lesions and 124 controls. Results: Patients with Basal Cell Skin Carcinoma (BCC) presented the combined GSTM1-GSTT1+ genotype more frequently (49.1%) than controls (29.8%) (Fisher test; p =0.04), conferring a 2.273 (Odds Ratio; 95% CI =1.199-4.308) higher risk for BCC. We were not able to find any other association between genotypes or between any genotype and the patients' clinical features. Conclusions: The GST profile may help identify Brazilian individuals at higher risk for BCC Antecedentes: La identificación de grupos en riesgo elevado es fundamental en la determinación de las estrategias preventivas para el cáncer de la piel, el maligno humano más común. Las especies reactivas destructivas del oxígeno producidas por UVA o los agentes carcinógenos químicos son metabolizadas por una serie de enzimas. Los polimorfismos de los genes que codifican para estas enzimas pueden producir las enzimas defectuosas con una capacidad disminuida de desintoxicar una amplia gama de agentes carcinógenos. Objetivo: Este estudio fue diseñado para comprobar las interacciones de la influencia y del potencial de varios polimorfismos de los genes que codificaban 4 enzimas importantes del antioxidante GST en la susceptibilidad al cáncer entre brasileños. Métodos: Comparamos los genotipos del mu del S-Transferase del Glutathione, de la theta, de pi y de Omega (GSTM1, GSTT1, GSTP1 y GSTO2) en un grupo de 102 lesiones de piel y de 124 controles. Resultados: Los pacientes con el carcinoma basocelular (BCC) presentaron el genotipo combinado de GSTM1-GSTT1+ más frecuente (49,1%) que los controles (29,8%) (Fisher test; p =0,04), confiriendo 2.273 (Odds Ratio 95% CI =1.199-4.308) un riesgo más alto para BCC. No encontramos ninguna otra asociación entre los genotipos o entre ningún genotipo y características clínicas de los pacientes. Conclusiones: Sugerimos que el perfil de GST pueda ayudar a identificar a individuos brasileños en un riesgo más alto para BCC

Background: p53 is a nuclear protein that exerts an important role in the negative control of cellular proliferation, as well as in masterminding signaling cascades important in DNA repair and/or apoptosis. Mutations of p53 have been reported with high frequency in many cancer types and are highly prevalent in poorly differentiated and undifferentiated thyroid carcinomas, but they are not found in benign tumors and are infrequent in well-differentiated cancer. Most mutations are located in exons 5-8 of the gene. Recently, a germline mutation in the seldom investigated exon 10, on codon 337 of p53 was described in Brazilian children who had adrenocortical tumors. Aim: To study codon 337 of exon 10 of p53 mutation in thyroid tumors. Material and methods: Seventy four thyroid tumors were studied (5 follicular carcinomas including 3 widely invasive, 22 papillary carcinomas including 6 tall cell variants, 11 follicular adenomas, 1 medullary carcinoma and 35 benign goiters). DNA was extracted from a central part of all tumors and contralateral normal thyroid tissue samples or blood from 38 of these patients. The products of PCR for exon 10 of p53 were examined by single strand conformation polymorphism (SSCP) analysis. We sequenced 2 samples suspected of presenting aberrant migrating bands and 3 additional PCR products from tumor samples with normal SSCP patterns but all were wild type. Results: In all samples studied, a wild type sequence was found. Conclusions: Exon 10 of p53 gene does not present mutations in thyroid tumors, suggesting that this mutation is specific of adrenocortical cancers. (Rev Méd Chile 2004; 132: 1513-6)

Orientador: Laura Sterian Ward Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas Resumo: Tanto a Doença de Graves (DG) como o Carcinoma Diferenciado da Tiroide (CDT) são patologias de etiologia multifatorial e envolvem uma interação entre meio ambiente e fatores genéticos de predisposição. Ambas apresentam nítida preferência pelo sexo feminino. Por isso, variações no metabolismo do estrógeno poderiam estar associadas a esta preferência, pois, seus metabólitos podem causar dano ao material genético. Os objetivos foram determinar a influência dos polimorfismos dos genes CYP17A1, HSD-17?1, CYP1A1, CYP1A2, CYP1B1, COMT e SULT1E1 no risco para a DG e para o CDT. Para tanto, foi estudado 282 pacientes com DG (234 mulheres e 48 homens; 39,80±11,69 anos), 292 pacientes com CDT (248 mulheres e 44 homens; 42,23±14,81 anos), comparados com 308 controles (246 mulheres e 62 homens; 36,86±12,95 anos). Para o estudo dos polimorfismos, utilizou-se a técnica TaqMan SNP Genotyping. Alterações polimórficas nos genes CYP17A1 (p=0,0421), CYP1A1 m1 (p=0,0328), CYP1A2*1F (p=0,0085), CYP1B1 códon 119 (p

Orientadores: Laura Sterian, Clarice Weiss Arns Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas Resumo: O Epstein-Barr Vírus (EBV) é um dos mais frequentes herpesvírus no mundo todo, cerca de 90% da população adulta já foram infectadas. Responsável por 2% de todas as neoplasias malignas no mundo, é considerado um agente carcinógeno humano. O vírus está relacionado com carcinoma de nasofaringe, câncer gástrico, linfomas de Burkitt e de Hodgkin e mais recentemente nosso grupo demonstrou a sua presença em alta carga viral em tumores malignos tireoidianos. O câncer de tireoide, por sua vez, é a neoplasia maligna endócrina mais comum em todo o mundo, sendo a quinta mais frequente entre as mulheres. Aumento da expressão nos oncogenes da família RAS (HRAS, NRAS e KRAS) são encontradas em diversos tumores humanos, e são frequentes no câncer de tireoide, alterações na expressão de TP53 também ocorrem, particularmente em tumores tireoidianos mais agressivos. O EBV é capaz de causar infecção latente e persistente no hospedeiro e interfere no metabolismo da célula hospedeira, desencadeando um processo tumorigênico. Sendo assim o objetivo deste trabalho foi avaliar a capacidade do Epstein-Barr Vírus em causar alterações morfológicas e moleculares em células tireoidianas neoplásicas das linhagens TPC-1, BCPAP e 8505C. Para tanto foi realizada a adaptação viral para análise de permissividade e efeito citopático nas linhagens celulares neoplásicas tireodianas, seguindo-se a cinética viral de 48 horas para análise da carga viral e da expressão dos oncogenes KRAS, NRAS, HRAS e de TP53. A comparação das células inoculadas pelo vírus com células controle não inoculadas mostrou que todas as linhagens celulares foram permissivas ao vírus, no entanto o vírus provocou efeito citopático, alterando a morfologia celular nas linhagens de TPC-1 e 8505C. Em relação à análise da expressão gênica observamos uma tendência à maior expressão de TP53 e NRAS nas linhagens TPC-1 e BCPAP inoculadas em relação ao grupo controle, e à maior expressão NRAS com diminuição da expressão de TP53 na linhagem 8505C. No entanto essa diferença não foi significativa (p>0,05). Concluímos que as linhagens células neoplásicas da tireoide são permissivas ao EBV e que o vírus é capaz de causar efeito citopático tanto nas linhagens celulares de carcinoma bem diferenciado como na de indiferenciado da tireoide. Demonstramos ainda que o EBV interfere na expressão gênica em linhagens celulares neoplásicas tireoidianas aumentando a expressão de oncogenes, principalmente KRAS e NRAS. Estes dados sugerem que EBV pode atuar na tumorigênese tireoidiana aumentando a expressão de oncogenes e interferindo na expressão de genes supressores tumorais Abstract: The Epstein-Barr Virus (EBV) is one of the most frequent herpes viruses worldwide, about 90% of the adult population have already been infected. Responsible for 2% of all malignancies in the world, it is a human carcinogen. The virus is related to nasopharyngeal carcinoma, gastric cancer, Burkitt and Hodgkin's lymphomas and more recently our group has demonstrated its presence in high viral load in malignant thyroid tumors. Thyroid cancer, in turn, is one of the most common endocrine malignancies in the world, a fifth most frequent among women. The hyperexpression of RAS family oncogenes (HRAS, NRAS and KRAS) occurs in several human tumors and it is frequent in thyroid cancer. Variations in TP53 expression also occur, particularly in more aggressive thyroid tumors. EBV is able to stablish latent and persistent infection in the host cell and does not interfere in its metabolism, triggering a tumorigenic process. Thus, we aimed to evaluate the ability of the virus to cause morphological and molecular changes in neoplastic thyroid cells of the TPC-1, BCPAP and 8505C lines. we aimed to evaluate the ability of the virus to cause morphological and molecular changes in neoplastic thyroid cell lines TPC-1, BCPAP and 8505C. Viral adaptation for the analysis of the cytopathic effect of the neoplastic thyroid cell lines was performed, followed by viral kinetics of 48 hours for viral load and genes expression analysis of KRAS, NRAS, HRAS and TP53. Comparison of the inoculated cells with non-inoculated control cells showed that all tumor cell lines were permissive to the virus; however, the virus caused cytopathic effect, altering the cell morphology only in the TPC-1 and 8505C. Related to gene expression analysis, we observed a tendency to greater expression of TP53 and NRAS in the TPC-1 and BCPAP lines inoculated in relation to the control group, and to the higher NRAS expression with decreased expression of TP53 in 8505C cell line. However, this difference was not significant (p> 0.05). We conclude that neoplastic thyroid cell lines are permissive to EBV and that the virus is capable of causing cytopathic effect in both well-differentiated and undifferentiated thyroid cell lines. We also demonstrated that EBV interferes in gene expression in thyroid neoplastic cell lines producing an increase in the expression of the oncogenes KRAS, NRAS and TP53 followed by the loss of expression of the tumor suppressor gene TP53. We suggest that these effects could be related to tumor progression Doutorado Clínica Médica Doutora em Ciências CAPES 1443334 FAEPEX 519.294

Camu-camu (Myrciaria dubia (Kunth) McVaugh) or Caçari is a native fruit species from the Amazon basin that has been noted worldwide for its richness in antioxidant compounds, such as ascorbic acid. Given this, EMBRAPA-RR and UFRR have been conducting several studies aiming to characterize the genetic variability and thus advance in the improvement of the species. However, there is still a lack of information generated using molecular marker techniques to abbreviate this process. Thus, the objective of this study was to identify, based on agronomic characteristics and molecular markers, individuals with different genetic profiles from the Embrapa-RR Camu-Camu Germplasm Collection. For this, the research was divided into three stages: 1) Firstly, we compared and modified protocols for DNA extraction from Roraima camu-camu. 2) Next, the genetic variability of 55 subsamples was analyzed using ISSR primers and 3) Finally, information from 53 subsamples was associated with agronomic data for more complete identification of the genotypes of the Collection. The process adaptations allowed us to establish a DNA extraction protocol for camu-camu according to the working conditions presented in Roraima, thus allowing to analyze the genetic variability present in the Germplasm Collection and to verify that AT08, AB08, AT13, BQ28, EV10, LR12 and IAB01 are the most genetically divergent. By combining the agronomic data with the molecular data, it was noted that more subsamples differed: AB08, AT08, AT10, AT13, BQ04, EV10, LR12, IAB04, BQ28, MU06 and LR05. Thus, it is extremely important to evaluate in the field the vegetative development and postharvest of these subsamples, being able to verify other agronomic attributes and to select genotypes that develop well in the field and at the same time maintain the present variation of the natural environment. O camu-camu (Myrciaria dubia (Kunth) McVaugh) ou caçari é uma espécie frutífera nativa da bacia Amazônica que têm se destacado mundialmente pela riqueza em compostos antioxidantes, como o ácido ascórbico. Diante disso, a EMBRAPA-RR e a UFRR vêm realizando diversos estudos com o objetivo de caracterizar a variabilidade genética e assim avançar no melhoramento da espécie. Entretanto, ainda há falta de informações geradas usando técnicas de marcadores moleculares que permitam abreviar esse processo. Sendo assim, objetivou-se com este estudo identificar com base em características agronômicas e marcadores moleculares indivíduos com perfis genéticos diferentes da Coleção de Germoplasma de camu-camu da Embrapa-RR. Para isso, a pesquisa foi dividida em três etapas: 1) Primeiramente foi comparado e modificado protocolos para extração de DNA do camu-camu de Roraima. 2) Em seguida, foi analisada a variabilidade genética de 55 subamostras por meio de iniciadores ISSR e 3) por último as informações moleculares de 53 subamostras foram associadas a dados agronômicos para identificação mais completa dos genótipos da Coleção. As adaptações nos processos permitiram estabelecer um protocolo de extração de DNA para o camu-camu de acordo com as condições de trabalho apresentadas em Roraima possibilitando assim, analisar a variabilidade genética presente na Coleção de Germoplasma e constatar que AT08, AB08, AT13, BQ28, EV10, LR12 e IAB01 são os mais divergentes geneticamente. Ao unir os dados agronômicos com os dados moleculares notouse que mais subamostras se diferenciaram, sendo elas: AB08, AT08, AT10, AT13, BQ04, EV10, LR12, IAB04, BQ28, MU06 e LR05. Dessa forma, é extremamente importante avaliar em campo o desenvolvimento vegetativo e a pós-colheita dessas subamostras, podendo verificar outros atributos agronômicos e selecionar genótipos que se desenvolvam bem no campo e ao mesmo tempo mantenham a variação presente do ambiente natural.