Improved protocol for Bst polymerase and reverse transcriptase production and application to a point-of-care diagnostics system.

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has raised awareness in the scientific community about the importance of being prepared for sanitary emergencies. Many measures implemented during the COVID pandemic are now being expanded to other applications. In the field of molecular and immunological diagnostics, the need to massively test the population worldwide resulted in the application of a variety of methods to detect viral infection. Besides gold standard reverse transcription quantitative polymerase chain reaction (RT-qPCR), the use of reverse transcription loop-mediated isothermal amplification (RT-LAMP) arose as an alternative and sensitive method to amplify and detect viral genetic material. We have used openly available protocols and have improved the protein production of RT-LAMP enzymes Bst polymerase and HIV-reverse transcriptase. To optimize enzyme production, we tested different protein tags, and we shortened the protein purification protocol, resulting in reduced processing time and handling of the enzymes and, thus, preserved the protein activity with high purity. The enzymes showed significant stability at 4 °C and 25 °C, over 60 days, and were highly reliable when used as a one-step RT-LAMP reaction in a portable point-of-care device with clinical samples. The enzymes and the reaction setup can be further expanded to detect other infectious diseases agents.

Hilab System Device in an Oncological Hospital: A New Clinical Approach for Point of Care CBC Test, Supported by the Internet of Things and Machine Learning.

The complete blood count (CBC) is a highly requested test that is generally restricted to centralized laboratories, which are limited by high cost, being maintenance-demanding, and requiring costly equipment. The Hilab System (HS) is a small, handheld hematological platform that uses microscopy and chromatography techniques, combined with machine learning (ML) and artificial intelligence (AI), to perform a CBC test. This platform uses ML and AI techniques to add higher accuracy and reliability to the results besides allowing for faster reporting. For clinical and flagging capability evaluation of the handheld device, the study analyzed 550 blood samples of patients from a reference institution for oncological diseases. The clinical analysis encompassed the data comparison between the Hilab System and a conventional hematological analyzer (Sysmex XE-2100) for all CBC analytes. The flagging capability study compared the microscopic findings from the Hilab System and the standard blood smear evaluation method. The study also assessed the sample collection source (venous or capillary) influences. The Pearson correlation, Student t-test, Bland-Altman, and Passing-Bablok plot of analytes were calculated and are shown. Data from both methodologies were similar (p > 0.05; r ≥ 0.9 for most parameters) for all CBC analytes and flagging parameters. Venous and capillary samples did not differ statistically (p > 0.05). The study indicates that the Hilab System provides humanized blood collection associated with fast and accurate data, essential features for patient wellbeing and quick physician decision making.

SARS-CoV-2 Delta and Omicron Variants Surge in Curitiba, Southern Brazil, and Its Impact on Overall COVID-19 Lethality.

Screening efforts and genomic surveillance are essential tools to evaluate the course of the COVID-19 pandemic and assist the public healthcare system in dealing with an increasing number of infections. For the analysis of COVID-19 cases scenarios in Curitiba, Paraná, Brazil, we performed a diagnosis of positive cases, coupled with genotyping, for symptomatic and asymptomatic members of the Federal University of Paraná. We achieved over 1000 samples using RT-qPCR for diagnosis. The posterior genotyping allowed us to observe differences in the spread of strains in Curitiba, Brazil. The Delta variant was not associated with an infection wave, whereas the rapid Omicron variant spread became dominant in less than one month. We also evaluated the general vaccination coverage in the state, observing a striking reduction in lethality correlated to the vaccinated fraction of the population; although lower lethality rates were not much affected by the Omicron variant wave, the same effect was not translated in the number of infections. In summary, our results provide a general overview of the pandemic's course in Paraná State and how there was reduction in lethality after a combination of multiple infection waves and a large-scale vaccination program.

Linkage strategy and vulnerabilities in the barriers to HIV/AIDS treatment for men who have sex with men. / Estratégia de linkagem e vulnerabilidades nas barreiras ao tratamento de HIV/Aids para homens que fazem sexo com homens.

This study aims to analyze the linkage to HIV/AIDS treatment among Men who have Sex with Men (MSM) of the project "A Hora é Agora" in Curitiba, Paraná, Brazil. The concept of vulnerability with its three axes, namely, the individual, social, and programmatic, was the theoretical framework. The barriers from testing up to the onset of the treatment were mapped through linkage registration and minutes from supervisory meetings. In the individual axis, the data revealed that the MSM struggled to address the HIV diagnosis, besides psychological problems that could cause the delay in starting treatment. Considering the social axis, stigma/discrimination was identified in the service at the Basic Health Care Network and within the families, delaying the disclosure of serology. Lastly, in the programmatic axis, the MSM found barriers in accessing the health services due to requests to repeat the HIV test, changing doctors due to poor service, and difficulties in conducting further tests, which adversely reflected on health care. In order to overcome these barriers, it is recommended not only a macro-structural work within this group, but also an investment in the micropolitics, which will enable a real change of attitude, continued care, and a stance related to approaches of care and the defense of life.

Este estudo tem por objetivo analisar a vinculação ao tratamento de HIV/Aids de Homens que fazem Sexo com Homens (HSH) no Projeto A Hora é Agora, na cidade de Curitiba, Paraná. O conceito de vulnerabilidade com seus três eixos: individual, social e programático foi considerado o marco teórico. Realizou-se levantamento das barreiras enfrentadas pelos sujeitos da testagem até o início do tratamento, por meio de registro da linkagem e atas das reuniões de supervisão. Os dados revelaram que, no plano individual, os HSH tiveram dificuldade em aceitar o diagnóstico de HIV, além de problemas psicológicos que podem ter acarretado na demora de início do tratamento. No eixo social, o estigma/discriminação foi identificado no atendimento nas Unidades Básicas de Saúde e na família, protelando a revelação da sorologia. Por fim, no eixo programático, os HSH encontraram entraves no acesso aos serviços de saúde em função: dos pedidos para repetirem o teste de HIV; mudança de médico pelo mau atendimento; e obstáculos na realização de outros exames, refletindo negativamente no cuidado da saúde. Para a superação dessas barreiras recomenda-se uma atuação não apenas macroestrutural frente a esse grupo, mas um investimento na micropolítica, possibilitando uma mudança real de atitude, cuidado contínuo e postura frente a abordagem do cuidador e a defesa da vida.

Estratégia de linkagem e vulnerabilidades nas barreiras ao tratamento de HIV/Aids para homens que fazem sexo com homens / Linkage strategy and vulnerabilities in the barriers to HIV/AIDS treatment for men who have sex with men

Resumo Este estudo tem por objetivo analisar a vinculação ao tratamento de HIV/Aids de Homens que fazem Sexo com Homens (HSH) no Projeto A Hora é Agora, na cidade de Curitiba, Paraná. O conceito de vulnerabilidade com seus três eixos: individual, social e programático foi considerado o marco teórico. Realizou-se levantamento das barreiras enfrentadas pelos sujeitos da testagem até o início do tratamento, por meio de registro da linkagem e atas das reuniões de supervisão. Os dados revelaram que, no plano individual, os HSH tiveram dificuldade em aceitar o diagnóstico de HIV, além de problemas psicológicos que podem ter acarretado na demora de início do tratamento. No eixo social, o estigma/discriminação foi identificado no atendimento nas Unidades Básicas de Saúde e na família, protelando a revelação da sorologia. Por fim, no eixo programático, os HSH encontraram entraves no acesso aos serviços de saúde em função: dos pedidos para repetirem o teste de HIV; mudança de médico pelo mau atendimento; e obstáculos na realização de outros exames, refletindo negativamente no cuidado da saúde. Para a superação dessas barreiras recomenda-se uma atuação não apenas macroestrutural frente a esse grupo, mas um investimento na micropolítica, possibilitando uma mudança real de atitude, cuidado contínuo e postura frente a abordagem do cuidador e a defesa da vida.

Abstract This study aims to analyze the linkage to HIV/AIDS treatment among Men who have Sex with Men (MSM) of the project "A Hora é Agora" in Curitiba, Paraná, Brazil. The concept of vulnerability with its three axes, namely, the individual, social, and programmatic, was the theoretical framework. The barriers from testing up to the onset of the treatment were mapped through linkage registration and minutes from supervisory meetings. In the individual axis, the data revealed that the MSM struggled to address the HIV diagnosis, besides psychological problems that could cause the delay in starting treatment. Considering the social axis, stigma/discrimination was identified in the service at the Basic Health Care Network and within the families, delaying the disclosure of serology. Lastly, in the programmatic axis, the MSM found barriers in accessing the health services due to requests to repeat the HIV test, changing doctors due to poor service, and difficulties in conducting further tests, which adversely reflected on health care. In order to overcome these barriers, it is recommended not only a macro-structural work within this group, but also an investment in the micropolitics, which will enable a real change of attitude, continued care, and a stance related to approaches of care and the defense of life.

Severe acute respiratory syndrome coronavirus 2 infection among healthcare workers in a tertiary public hospital in Curitiba, Brazil.

BACKGROUND: We aimed to describe the clinical characteristics of coronavirus disease 2019 (COVID-19) among healthcare workers (HCWs) in Curitiba, Brazil. METHODS: Upper respiratory samples from 1077 HCWs were tested for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using reverse transcription polymerase chain reaction from June 16, 2020 to December 9, 2020. RESULTS: Overall, 32.7% of HCWs were infected. The positivity rates in symptomatic and asymptomatic HCWs were 39.2% and 15.9%, respectively. Hospital departments categorized as high-risk for exposure had the highest number of infected HCWs. CONCLUSIONS: Early diagnosis and isolation of infected HCWs remain key in controlling SARS-CoV-2 transmission because HCWs in close contact with COVID-19 patients are more likely to be infected than those who are not.

Severe acute respiratory syndrome coronavirus 2 infection among healthcare workers in a tertiary public hospital in Curitiba, Brazil

ABSTRACT BACKGROUND: We aimed to describe the clinical characteristics of coronavirus disease 2019 (COVID-19) among healthcare workers (HCWs) in Curitiba, Brazil. METHODS: Upper respiratory samples from 1077 HCWs were tested for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using reverse transcription polymerase chain reaction from June 16, 2020 to December 9, 2020. RESULTS: Overall, 32.7% of HCWs were infected. The positivity rates in symptomatic and asymptomatic HCWs were 39.2% and 15.9%, respectively. Hospital departments categorized as high-risk for exposure had the highest number of infected HCWs. CONCLUSIONS: Early diagnosis and isolation of infected HCWs remain key in controlling SARS-CoV-2 transmission because HCWs in close contact with COVID-19 patients are more likely to be infected than those who are not.

Small vessel occlusion and syphilis in patients with first-ever ischemic stroke.

BACKGROUND: Syphilis is an endemic disease, particularly in low- and middle-income countries, with vascular involvement in large vessels (aortitis), but no clear relationship with stroke patients, except for those who presented with meningovascular neurosyphilis. OBJECTIVE: To investigate the relationship between a positive history of syphilis determined by serological testing and ischemic stroke etiology, particularly small vessel disease (SVD). METHODS: In total, 269 first-ever ischemic stroke patients admitted to the stroke unit were tested for syphilis. Patients with neurosyphilis were excluded. All patients were classified according to the ASCOD phenotyping as SVD - when SVD was the potential causal mechanism (S1) - or non-SVD - when SVD was uncertain (S2), unlike (S3), or not detected (S0). RESULTS: Syphilis was positive in 32 (12%) patients. When comparing patients with positive and negative serology, the only significant difference was SVD as the causal mechanism (S1) in patients with positive results: 9 (28%) vs. 22 (9%), p<0.01. CONCLUSION: The current study showed that the frequency of positive syphilis serological test was higher in patients with first-ever ischemic stroke and SVD as the potential causal mechanism. This finding could be related to the endothelial dysfunction occurring in syphilis.