SUMMARY OBJECTIVE: The aim of this study was to examine the impact of symptom-based screening on the prevalence and outcomes of neonatal coronavirus disease 2019 in pregnant women admitted for delivery. METHODS: A retrospective observational study was conducted from June to August 2020 at Gonzaga Mota of Messejana Hospital, Fortaleza, CE, Brazil. All pregnant women were screened for coronavirus disease 2019 based on symptoms. Reverse transcription-polymerase chain reaction or immunology assays for severe acute respiratory syndrome coronavirus 2 were performed when a patient reported a symptom. All newborns of symptomatic patients were submitted for Reverse transcription-polymerase chain reaction. Newborns were divided into groups according to the Reverse transcription-polymerase chain reaction results to identify the relationship between maternal symptoms and neonatal coronavirus disease 2019. RESULTS: A total of 55 (55/1,026, 5.4%) and 50 (50/1,026, 4.8%) pregnant women reported symptoms and had a positive confirmatory test, respectively. The most common symptom of coronavirus disease 2019 among the pregnant women with positive confirmatory test was cough (n=23, 46%). Seven newborns (7/50, 14%) of symptomatic mothers had positive Reverse transcription-polymerase chain reaction. Upon birth, no newborn had serious complications. CONCLUSION: Universal screening of pregnant women admitted for delivery can reduce the perinatal transmission of coronavirus disease 2019. Symptom-based screening can be an alternative for regions with a low prevalence of the disease where a better allocation of financial resources is necessary.

Abstract Zika fever is a viral infection of great relevance in public health, especially in tropic regions, in which there is a predominance of mosquitoes of the genus Aedes, vectors of the disease. Microcephaly in neonatal children and Guillain-Barré syndrome in adults can be caused by the action of the Zika virus (ZIKV). Non-structural proteins, such as NS2B, NS3 and NS5, are important pharmacological targets, due to their action in the life cycle. The absence of anti-Zika drugs raises new research, including prospecting for natural products. This work investigated the in silico antiviral activity of bixin and six other derived molecules against the Zika viral proteins NS2B-NS3 and NS5. The optimized structure was subjected to molecular docking to characterize the interaction between bixinoids and ZIKV non-structural proteins, where significant interactions were observed with amino acid residues in the catalytic site in each enzyme. These results suggest that bixin and ethyl bixin has the potential to interfere with the enzymatic activity of NS2B, NS3 and NS5, thus being an indication of being a promising anti-Zika agent.

Abstract INTRODUCTION: Quantitative reverse transcription polymerase chain reaction (RT-qPCR) can detect the severe acute respiratory syndrome Coronavirus-2 (SARS-CoV-2) in a highly specific manner. However, a decrease in the specificity of PCR assays for their targets may lead to false negative results. METHODS: Here, 177 high-coverage complete SARS-CoV-2 genome sequences from 13 Brazilian states were aligned with 15 WHO recommended PCR assays. RESULTS: Only 3 of the 15 completely aligned to all Brazilian sequences. Ten assays had mismatches in up to 3 sequences and two in many sequences. CONCLUSION: These results should be taken into consideration when using PCR-based diagnostics in Brazil.

The present study aimed to review literature on studies of dengue cases conducted over 30 years in the state of Ceará. Between November 2015 and January 2016, articles published in Portuguese and English in 7 databases were searched using keywords and a Boolean operator. A total of 191 articles were identified in the databases; 133 were excluded according to the exclusion criteria, and 58 were included in the study. Of the 58 articles analyzed, 6 reported data from Brazil; including the Northeast region and the state of Ceará; 41 reported data for only the city of Fortaleza; 7 reported data for the state of Ceará; 4 reported data for cities in the interior of the state; and 3 included only children. The studies adopted different approaches and focused on different aspects of the disease. Study outcomes included the identification of serological, epidemiological, clinical, and laboratory characteristics; potential larvicides and biological predators of mosquitoes; potential antiviral agents; vector density characteristics; and educational dengue prevention and control strategies. Additionally, one vaccine trial was included. Although studies on dengue in the state of Ceará are scarce, they are encompassing, including several lines of research, and the number of studies and reports on dengue in the state of Ceará continues to increase.

Abstract INTRODUCTION: Early diagnosis of human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) can decrease transmission and significantly affect morbidity and mortality; however, Brazil still confronts the reality of late HIV diagnosis. METHODS: Medical records of 284 HIV-positive patients were reviewed in this cross-sectional study. RESULTS: Of all patients, 28% were diagnosed in the context of health assessments, whereas 27% were symptomatic at diagnosis. Early HIV infection (Group 1) was diagnosed in 60.2% of participants. They were younger than those with late diagnosis (Group 2) (p = 0.002). CONCLUSIONS: These findings highlight the need for strategies to increase HIV testing in asymptomatic individuals and older patients.

Introducción. El dengue y la leptospirosis son dos enfermedades febriles de gran importancia clínica y epidemiológica en Brasil, y presentan una similitud sintomatológica significativa, lo cual dificulta el diagnóstico clínico. Objetivos. Hacer el diagnóstico diferencial para leptospirosis en pacientes con sospecha clínica de dengue. Materiales y métodos. En este estudio, 86 pacientes con sospecha clínica de dengue fueron sometidos a examen para el diagnóstico de dengue mediante pruebas de virología y serología (RT-PCR, prueba de inmunocromatografía NS1 y ELISA-NS1) y pruebas para la detección de IgM (prueba rápida IgM/IgG y ELISA-IgM). En los mismos pacientes también se evaluó la presencia de leptospirosis mediante la prueba rápida IgG/IgM (Bioeasy ® ) y ELISA IgM-Leptospira (PanBio ® ) . Resultados. De los 86 pacientes, 48 (55,8 %) fueron positivos para dengue en, por lo menos, una de las pruebas y 5 (7,35 %) presentaron reacciones positivas para leptospirosis. Conclusión. Durante las epidemias de dengue, otras infecciones y entre ellas la leptospirosis, pueden dar lugar a confusiones cuando se el diagnóstico se hace únicamente con base en criterios inespecíficos clínicos y de laboratorio.

Introduction: Dengue and leptospirosis are two febrile illnesses of great clinical and epidemiological importance in Brazil. Their significant degree of symptomatic similarity makes clinical diagnosis difficult. Objective: To diagnose leptospirosis differentially in patients with clinically suspected dengue. Materials and methods: In this study, 86 patients with clinically suspected dengue underwent virological and serological diagnostic evaluations for dengue (reverse transcriptase polymerase chain reaction, NS1 immunochromatographic test, and NS1 enzyme-linked immunosorbent assay, ELISA), as well as tests to detect immunoglobulin M (IgM; IgM/IgG Rapid Test and IgM ELISA). The same patients were subsequently evaluated for leptospirosis using Rapid Test IgG/IgM (Bioeasy ® ) and Leptospira IgM ELISA (PanBio ® ). Results: Of the 86 patients, 48 (55.8%) had positive results for dengue in at least one of the tests and five (7.35%) showed positive reactions for leptospirosis. Conclusion: During dengue epidemics, this disease may be misdiagnosed as other infections, including leptospirosis, when diagnosis is based on nonspecific clinical and laboratory criteria alone.

RESUMO A leptospirose é doença febril tipicamente subestimada em todo o mundo, principalmente em áreas que a dengue se apresenta de forma endêmica. Desta forma, há limitações importantes na compreensão do número de casos de leptospirose nessas áreas, o que proporciona maior subnotificação. Neste estudo, apresentamos estimativa de possíveis casos de leptospirose a partir de casos de dengue-símile na cidade de Fortaleza, nordeste do Brasil, durante os anos de 2008, 2010 e 2012. Os pacientes foram investigados para dengue e leptospirose utilizando testes imunoezimáticos para detecção do anticorpo, da classe IgM, específicos para cada patologia. Entre os casos suspeitos de dengue, mas que não apresentaram resultado laboratorial positivo, 10,8%; 19,2% e 30,8% foram confirmados como leptospirose nos anos de 2008, 2010 e 2012; respectivamente. Considerando os casos notificados pela vigilância de dengue e que foram, posteriormente, descartados, baseados nos resultados dos testes laboratoriais, estimamos que o número atual de casos de leptospirose pode ser de 26 a 49 vezes mais do que o detectado e notificado pelos serviços de saúde. Além disso, acreditamos que aproximadamente 20% dos casos de dengue-símile podem ser de leptospirose, em áreas onde as duas doenças ocorram de forma endêmica.

SUMMARY Leptospirosis is a febrile disease with a typically underestimated global incidence, especially in regions where dengue is endemic. Therefore, it is difficult to accurately determine the number of leptospirosis cases in these areas, which contributes to significant under-reporting this disease. In this study, we estimated the number of possible leptospirosis cases among dengue-like cases that were reported during 2008, 2010, and 2012 in the city of Fortaleza, northeast Brazil. Patients were evaluated for dengue and leptospirosis using immunoenzymatic tests for IgM antibodies that were specific to each pathogen. Among the suspected cases of dengue that resulted as negative in laboratory tests, 10.8% (2008), 19.2% (2010), and 30.8% (2012) were confirmed to be leptospirosis. Considering the cases reported by the surveillance authority as dengue that were subsequently discarded based on the laboratory test results, we estimate that the number of actual leptospirosis cases may be 26 to 49 times higher than those diagnosed and reported by the Health Services. Furthermore, we believe that approximately 20% of dengue-like cases may be leptospirosis cases in areas where the two diseases are endemic.

While human immunodeficiency virus (HIV)-1 chemokine co-receptors 5 tropism and the GWGR motif in the envelope third variable region (V3 loop) have been associated with a slower disease progression, their influence on antiretroviral response remains unclear. The impact of baseline V3 characteristics on treatment response was evaluated in a randomised, double blind, prospective cohort study with patients initiating highly active antiretroviral therapy with lopinavir or efavirenz plus azithothymidine/3TC (1:1) over 48 weeks. Similar virological and immunological responses were observed for both treatment regimens. The 43 individuals had a mean baseline CD4 T cell count of 119 cells/mm³ [standard deviation (SD) = 99] and a mean viral load of 5.09 log10 copies/mL (SD = 0.49). The GWGR motif was not associated with a CD4 T cell response, but predicted R5 tropism by the geno2pheno[clinical20%] algorithm correlated with higher CD4 T cell levels at all monitoring points (p < 0.05). Moreover, higher false-positive rates (FPR) values from this analysis revealed a strong correlation with CD4 T cell recovery (p < 0.0001). Transmitted drug resistance mutations, documented in 3/41 (7.3%) cases, were unrelated to the assigned antiretroviral regimen and had no impact on patient outcomes. In conclusion, naÏve HIV-1 R5 infected patients exhibited higher CD4 T cell counts at baseline; this difference was sustained throughout therapy. The geno2pheno[clinical] option FPR positively correlated with CD4 T cell gain and may be useful in predicting CD4 T cell recovery.

AIM: The objective of this study was to correlate laboratory tests during the evolution of dengue fever, comparing frequencies between the different clinical forms in order to use test results to predict the severity of the disease. METHODS: This is an observational, descriptive and retrospective study of 154 patients with clinical and serological diagnoses of dengue fever who, in the period from January to May 2008, were admitted in a tertiary state hospital in the city of Fortaleza that is a referral center for infectious diseases. The patients were allocated to two groups according to age: under 15 years old (n = 66) and 15 years or older (n = 88). The tests analyzed were blood count, platelet count, and serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) concentrations. RESULTS: Thrombocytopenia and elevated transaminases were observed in patients with classic dengue fever. The main laboratory abnormalities found in dengue hemorrhagic fever were thrombocytopenia, hemoconcentration and elevated transaminases, similar to severe dengue with the exception of hemoconcentration. Most laboratory abnormalities started on the 3rd day but were more evident on the 5th day with restoration of values by the 11th day; this was more prominent in under 15-year-olds and with the more severe clinical forms. CONCLUSIONS: These results are relevant in assessing the disease because they can be used as markers for more severe forms and can help by enabling the adaptation of the therapeutic conduct to the needs of individual patients.

A reação em cadeia da polimerase (PCR) tem fornecido diagnóstico de material de arquivo, mas alguns métodos de fixação, tais como formalina, provocam danos ao DNA e subsequentemente afetam sua análise, particularmente tecidos embebidos em parafina. A PCR é conhecida pela sua alta especificidade e sensibilidade, embora algumas dificuldades ocorram quando o material utilizado foi fixado em formalina e embebido em parafina. Isso não se deve somente pela formação de cross-linkings com proteínas, a qual aumenta com o maior tempo de fixação, mas também pelo dano direto que a formalina causa no DNA. PCR foi usada para analisar placenta e órgão fetais de 34 amostras com suspeita de infecção pelo Parvovírus B19 (PB19). Não foi possível amplificar o DNA do PB19 usando nested-PCR, provavelmente devido ao tamanho do amplicon gerado com o primeiro passo dos primers. Adequamos o problema utilizando somente o segundo par de primers e pudemos observar que das 34 amostras, duas eram positivas para PCR (5,9%). Entretanto, a PCR dos órgãos fetais foi negativa em um de dois casos. Observamos também uma relação negativa entre a espessura do corte dos materiais com a positividade das amostras. Em conclusão, embora a PCR seja altamente específica e sensível em amostras a fresco ou idealmente fixadas, uma padronização cuidadosa para análise com a PCR é necessária quando se utiliza tecidos fixados em formalina e embebidos em parafina utilizando primers que requerem menor fragmento de DNA para a amplificação.

The polymerase chain reaction (PCR) has provided diagnosis of archival material, but some fixation methods such as formalin damage DNA and, subsequently, affect PCR analysis, particularly paraffin-embedded tissues. PCR is known due to its high specificity and sensitivity, although some difficulties arise when formalinfixed and paraffin-embedded tissue is used. Not only does this occur due to protein cross-linking, which increases with longer fixation time, but it also happens due to the direct damage that formalin causes in the DNA itself. PCR was used to analyze placenta and fetal organs from 34 samples with suspected Parvovirus B19 infection. It was not possible to amplify Parvovirus B19 DNA using nested-PCR, probably due to the size of the amplicon generated with the first set of primers. We approached this problem by using only the second set of primers. Two out of 34 tissue samples (5,9%) were positive by PCR. However, PCR performed on corresponding fetal organs was negative in one of the two. We also observed a negative relation between the thickness of the tissue fragment and the positivity of the samples. In conclusion, although PCR is highly specific and sensitive in fresh or ideally fixed material, a careful standardization of PCR assays is necessary when using formalin fixed paraffin-embedded tissues by applying primers that require smaller DNA fragments for amplification.

O parvovírus B19 foi detectado em 1975 e desde sua descoberta tem se mostrado um agente infeccioso importante em seres humanos, cujo diagnóstico pode ser feito pelo exame histológico através do encontro de inclusão nuclear em tecidos fetais ou placentários. No entanto, estas células podem ser escassas ou não apresentarem características típicas, dificultando o diagnóstico. Analisamos placentas e órgãos fetais de 34 casos de hidropisia fetal não-imune corados com Hematoxilina e Eosina (HE) e submetidos à reação em cadeia da polimerase (PCR) e imuno-histoquímica (IH). Em dois casos (5,9%) houve positividade na placenta pelo HE, IH e PCR. No entanto, PCR dos órgãos fetais foi negativa em um destes casos enquanto que a identificação pôde ser feita por IH e histologia. Concluímos que a freqüência do parvovírus B19 é similar à literatura e a reação IH foi o melhor método de detecção, com identificação mais específica e segura, permitindo identificação citoplasmática, o que não é possível pelo exame histopatológico. A PCR pode apresentar falsa negatividade, provavelmente pela fixação, não identifica as células e é mais dispendiosa. Embora mais específica e sensível em material a fresco ou idealmente fixado isto não ocorre com tecidos fixados em formalina e embebidos em parafina, freqüentemente os únicos disponíveis.

Parvovirus B19 infection was first discovered in 1975 and it is implicated in fetal death from hydrops fetalis the world over. Diagnosis is usually made through histological identification of the intranuclear inclusion in placenta and fetal organs. However, these cells may be scarce or uncharacteristic, making definitive diagnosis difficult. We analyzed histologically placentas and fetal organs from 34 cases of non-immune hydrops fetalis, stained with Hematoxylin and Eosin (HE) and submitted to immunohistochemistry and polymerase chain reaction (PCR). Of 34 tissue samples, two (5.9%) presented typical intranuclear inclusion in circulating normoblasts seen in Hematoxylin and Eosin stained sections, confirmed by immunohistochemistry and PCR. However, PCR of fetal organs was negative in one case in which the placenta PCR was positive. We concluded that parvovirus B19 infection frequency is similar to the literature and that immunohistochemistry was the best detection method. It is highly specific and sensitive, preserves the morphology and reveals a larger number of positive cells than does HE with the advantage of showing cytoplasmic and nuclear positivity, making it more reliable. Although PCR is more specific and sensitive in fresh or ideally fixed material it is not so in formalin-fixed paraffin-embedded tissues, frequently the only one available in such cases.