Recent dynamics in Neisseria gonorrhoeae genomic epidemiology in Brazil: antimicrobial resistance and genomic lineages in 2017-20 compared to 2015-16.

OBJECTIVES: Regular quality-assured WGS with antimicrobial resistance (AMR) and epidemiological data of patients is imperative to elucidate the shifting gonorrhoea epidemiology, nationally and internationally. We describe the dynamics of the gonococcal population in 11 cities in Brazil between 2017 and 2020 and elucidate emerging and disappearing gonococcal lineages associated with AMR, compare to Brazilian WGS and AMR data from 2015 to 2016, and explain recent changes in gonococcal AMR and gonorrhoea epidemiology. METHODS: WGS was performed using Illumina NextSeq 550 and genomes of 623 gonococcal isolates were used for downstream analysis. Molecular typing and AMR determinants were obtained and links between genomic lineages and AMR (determined by agar dilution/Etest) examined. RESULTS: Azithromycin resistance (15.6%, 97/623) had substantially increased and was mainly explained by clonal expansions of strains with 23S rRNA C2611T (mostly NG-STAR CC124) and mtr mosaics (mostly NG-STAR CC63, MLST ST9363). Resistance to ceftriaxone and cefixime remained at the same levels as in 2015-16, i.e. at 0% and 0.2% (1/623), respectively. Regarding novel gonorrhoea treatments, no known zoliflodacin-resistance gyrB mutations or gepotidacin-resistance gyrA mutations were found. Genomic lineages and sublineages showed a phylogenomic shift from sublineage A5 to sublineages A1-A4, while isolates within lineage B remained diverse in Brazil. CONCLUSIONS: Azithromycin resistance, mainly caused by 23S rRNA C2611T and mtrD mosaics/semi-mosaics, had substantially increased in Brazil. This mostly low-level azithromycin resistance may threaten the recommended ceftriaxone-azithromycin therapy, but the lack of ceftriaxone resistance is encouraging. Enhanced gonococcal AMR surveillance, including WGS, is imperative in Brazil and other Latin American and Caribbean countries.

Prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, and Mycoplasma genitalium and risk factors among pregnant women in Brazil: Results from the national molecular diagnosis implementation project.

BACKGROUND: Sexually transmitted infections (STIs) are a public health problem. The aim of the present study was to assess the prevalence and risk factors associated with at least one STI (Chlamydia trachomatis [CT], Neisseria gonorrhoeae [NG], Trichomonas vaginalis [TV], and Mycoplasma genitalium [MG]) in Brazil. METHODS: A cross-sectional study was conducted using secondary data from the pilot implementation of the National Service for molecular diagnosis of CT, NG, TV, and MG in pregnancy. We obtained Ministry of Health surveillance data from the implementation project. Data encompassing pregnant women aged 15-49 years from public antenatal clinics in Brazil in 2022 were included. RESULTS: A total of 2728 data of pregnant women were analyzed. The prevalence of at least one infection was 21.0% (573), with the highest prevalence in the Southeast region (23.3%) and the lowest in the Center-West region (15.4%). The prevalence of CT was 9.9% (270), NG 0.6% (16), TV 6.7% (184), and MG 7.8% (212). Factors associated with any infection were from 15 to 24 years (AOR = 1.93; 95% CI: 1.58-2.35); reported family income up to US$400 (AOR = 1.79; 95% CI: 1.03-3.34); declared not living maritally with their partners (AOR = 1.90, 95% CI: 1.52-2.37) and had more than one sexual partner in their lifetime (AOR = 2.09, 95% CI: 1.55-2.86). CONCLUSION: This study showed a high prevalence of at least one STI among pregnant women in Brazil, particularly among younger women. It also provides up-to-date national data on CT, NG, TV, and MG infections in this population. These findings underscore the importance of enhancing access to STI screening for young pregnant women within the Brazilian public health system.

Aetiological molecular identification of sexually transmitted infections that cause urethral discharge syndrome and genital ulcer disease in Brazilian men: a nationwide study.

BACKGROUND: Little is known about the aetiology of urethral discharge syndrome (UDS) and genital ulcer disease (GUD) in Brazil due to limited access to laboratory tests and treatment based mainly on the syndromic approach. OBJECTIVES: To update Brazilian treatment guidelines according to the current scenario, the first nationwide aetiological study for UDS and GUD was performed. METHODS: Male participants with urethral discharge (UD) and/or genital ulcer (GU) reports were enrolled. Sample collection was performed by 12 sentinel sites located in the five Brazilian regions. Between 2018 and 2020, 1141 UD and 208 GU samples were collected in a Universal Transport Medium-RT (Copan). A multiplex quantitative PCR kit (Seegene) was used to detect UD: Chlamydia trachomatis (CT), Mycoplasma genitalium (MG), M. hominis (MH), Neisseria gonorrhoeae (NG), Trichomonas vaginalis (TV), Ureaplasma parvum (UP), U. urealyticum (UU) and another kit to detect GU: cytomegalovirus (CMV), Haemophilus ducreyi (HD), herpes simplex virus type 1 (HSV1), herpes simplex virus type 2 (HSV2), lymphogranuloma venereum (LGV), Treponema pallidum (TP) and varicella-zoster virus (VZV). RESULTS: In UD samples, the frequency of pathogen detection was NG: 78.38%, CT: 25.6%, MG: 8.3%, UU: 10.4%, UP: 3.5%, MH: 3.5% and TV: 0.9%. Coinfection was assessed in 30.9% of samples, with 14.3% of NG/CT coinfection. The most frequent pathogen identified in GU was HSV2, present in 40.8% of the samples, followed by TP at 24.8%, LGV and CMV at 1%, and HSV1 at 0.4%. Coinfection of TP/HSV2 was detected in 4.4% of samples. VZV and HD were not detected. In 27.7% of the GU samples, no pathogen was detected. CONCLUSION: This study provided the acquisition of unprecedented data on the aetiology of UDS and GUD in Brazil, demonstrated the presence of a variety of pathogens in both sample types and reaffirmed the aetiologies known to be most prevalent globally.

Recent HIV infection and annualized HIV incidence rates among sexual and gender minorities in Brazil and Peru (ImPrEP seroincidence study): a cross-sectional, multicenter study.

Background: HIV incidence estimation is critical for monitoring the HIV epidemic dynamics and the effectiveness of public health prevention interventions. We aimed to identify sexual and gender minorities (SGM) with recent HIV infections, factors associated with recent HIV infection, and to estimate annualised HIV incidence rates. Methods: Cross-sectional multicentre study in HIV testing services in Brazil and Peru (15 cities). Inclusion criteria: 18+ years, SGM assigned male at birth, not using pre-/post-exposure prophylaxis. We identified recent HIV infection using the Maxim HIV-1 LAg-Avidity EIA assay as part of a recent infection testing algorithm (RITA). Annualized HIV incidence was calculated using the UNAIDS/WHO incidence estimator tool. Multivariable logistic regression models were used to estimate factors associated with recent HIV infection. Trial registration: NCT05674682. Findings: From 31-Jan-2021 to 29-May-2022, 6899 individuals participated [Brazil: 4586 (66.5%); Peru: 2313 (33.5%)]; 5946 (86.2%) cisgender men, 751 (10.9%) transgender women and 202 (2.9%) non-binary/gender diverse. Median age was 27 (IQR: 23-34) years. HIV prevalence was 11.4% (N = 784/6899); 137 (2.0%) SGM were identified with recent HIV infection. The overall annualized HIV incidence rate was 3.88% (95% CI: 2.86-4.87); Brazil: 2.62% (95% CI: 1.78-3.43); Peru: 6.69% (95% CI: 4.62-8.69). Participants aged 18-24 years had higher odds of recent HIV infection compared to those aged 30+ years in both countries. Interpretation: Our results highlight the significant burden of HIV epidemic among SGM in large urban centres of Brazil and Peru. Public health policies and interventions to increase access to effective HIV prevention methods such as PrEP are urgently needed in Latin America. Funding: Unitaid, WHO (Switzerland), Ministry of Health from Brazil and Peru.

Brazilian Clinical Practice Guidelines for Sexually Transmitted Infections That Cause Urethral Discharge: Assessment According to the AGREE II and Critical Analysis Based on the WHO Recommendations.

BACKGROUND: Sexually transmitted infections (STI) can have severe consequences. In Brazil, case management is recommended by the Clinical Protocol and Therapeutical Guidelines for Comprehensive Care for People with STIs (PCDT-IST). This study assessed the quality of PCDT-IST (2021) and reviewed the main recommendations for the management of STI that cause urethral discharge compared with the World Health Organization (WHO) STI Guidelines. METHODS: The PCDT-IST (2021) quality was independently assessed by 4 appraisers using the Appraisal of Guidelines Research and Evaluation instrument, version II (AGREE II). The PCDT-IST (2021) and the WHO Guidelines for the Management of Symptomatic STI (2021) were compared considering 14 different assessment domains. RESULTS: The PCDT-IST (2021) scores in the AGREE II domains were: Rigor of Development (58%), Applicability (35%), Editorial Independence (38%), Scope and Purpose (78%), Stakeholder Involvement (74%), and Clarity and Presentation (82%). The overall score was 67%, and all appraisers recommended the Brazilian guideline. Regarding the PCDT-IST (2021) and the WHO STI Guidelines (2021) comparation, 10 domains would be relevant for further reviewing the Brazilian recommendations: Diagnostic tests; Etiological approach; Treatment for recurrent urethral discharge; Treatment for urethritis without etiological agent identification; Treatment for gonococcal urethritis; Treatment for chlamydial urethritis; Retreatment for gonococcal infections; Treatment for Mycoplasma genitalium urethritis; Treatment for Trichomonas vaginalis urethritis; 10. Flowcharts. CONCLUSIONS: The PCDT-IST (2021) has a reasonable degree of quality. However, the domains of Applicability, Rigor of Development, and Editorial Independence must be better ensured. The guidelines comparison will help to select key topics that should be addressed with priority in the following national STI guidelines updates.

National surveillance of Neisseria gonorrhoeae antimicrobial susceptibility and epidemiological data of gonorrhoea patients across Brazil, 2018-20.

Objectives: To (i) describe the nationwide antimicrobial susceptibility of Neisseria gonorrhoeae (NG) isolates cultured across Brazil in 2018-20 and compare it with NG antimicrobial resistance data from 2015-16, and (ii) present epidemiological data of the corresponding gonorrhoea patients in 2018-20. Methods: Twelve representative sentinel sites cultured NG isolates from men with urethral discharge. Susceptibility to eight antimicrobials was examined using agar dilution method, according to WHO standards. The consenting participants were invited to provide epidemiological data. Results: In total, 633 NG isolates (one isolate per participant) were analysed, and 449 (70.9%) questionnaires were answered. Heterosexual (68.2%) and homosexual (23.1%) sexual orientations were common, and most prevalent types of unprotected sexual intercourse were vaginal insertive (69.9%), oral giving (56.6%) and anal insertive (47.4%). The levels of in vitro NG resistance to ciprofloxacin, tetracycline, benzylpenicillin, azithromycin, cefixime, gentamicin, spectinomycin and ceftriaxone were 67.3%, 40.0%, 25.7%, 10.6%, 0.3%, 0%, 0% and 0%, respectively. Compliance with the recommended first-line ceftriaxone 500 mg plus azithromycin 1 g therapy was high (90.9%). Conclusions: Compared with 2015-16, ciprofloxacin resistance has remained high and azithromycin and cefixime resistance rates have increased in Brazil. Resistance remained lacking to ceftriaxone, gentamicin and spectinomycin, which all are gonorrhoea treatment options. The increasing azithromycin resistance in Brazil and internationally may threaten the future use of azithromycin in dual regimens for treatment of gonorrhoea. Consequently, continued and enhanced quality-assured surveillance of gonococcal AMR, and ideally also treatment failures and including WGS, is imperative in Brazil and worldwide.

Prevalence of syphilis and associated factors in the adult population treated at healthcare facilities in Vitória (ES), Brazil / Prevalência e fatores associados à sífilis em adultos atendidos em unidades de saúde de Vitória (ES), Brasil

Introduction: There are few population-sampling studies on the prevalence of syphilis in Brazil. Objectives: We aim to determine the seroprevalence of syphilis and identify factors associated with the infection in adult patients observed at six regional healthcare facilities in Vitória city, state of Espírito Santo, Brazil. Methods: A cross-sectional study was conducted between September 2010 and December 2011. For individuals included in the study, a Venereal Disease Research Laboratory (VDRL) test and two treponemal tests (immunochromatographic and IgG ELISA assays) were performed. Demographic data, history of sexually transmitted infections, and behavioral data were collected. Results: Of the 1,502 subjects included in the study, 47% were men and 53% were women. The mean age was 41.63±14.57 years. The prevalence of syphilis was (0.9%; 95%CI 0.4­1.3) when a diagnosis of syphilis was considered with VDRL titers equal to or greater than 1:8. However, the prevalence was higher (2.8%) when a positive VDRL test, regardless of the titer, was considered. A multivariate analysis showed a significant association between syphilis and homosexual or bisexual behavior [OR=6.80; 95%CI 1.00­46.20], prior history of sexually transmitted infection [OR=16.30; 95%CI 3.61­73.41], the presence of a tattoo [OR=6.21; 95%CI 1.49­ 25.84], and cocaine use [OR=6.80; 95%CI 1.15­40.30]. The prevalence of positive treponemal test was 10.4%. Conclusion: The seroprevalence of active syphilis in this population was similar to that observed in other populational studies in Brazil. The high prevalence of positive treponemal tests may be due to the positive serological memory of a cured infection, but the results may also be due to cases of early or late syphilis that were not detected by the VDRL test.

Introdução: Existem poucos estudos em amostras populacionais sobre a prevalência da sífilis no Brasil. Objetivos: Determinar a soroprevalência de sífilis e identificar fatores associados à infecção em pacientes adultos atendidos nas unidades das seis regiões de saúde do Município de Vitória, Estado do Espírito Santo. Métodos: Foi realizado um estudo transversal entre setembro de 2010 e dezembro de 2011. Para os indivíduos incluídos no estudo, foram realizados o teste Venereal Disease Research Laboratory (VDRL) e dois testes treponêmicos (imunocromatográfico e IgG ELISA). Foram coletados dados demográficos, histórico de infecções sexualmente transmissíveis e dados comportamentais. Resultados: Dos 1.502 indivíduos incluídos no estudo, 47% eram homens e 53% eram mulheres. A média de idade foi de 41,63±14,57 anos. A prevalência de sífilis foi de 0,9% (IC95% 0,4­1,3) quando considerado diagnóstico de sífilis com títulos de VDRL iguais ou superiores a 1:8. Porém, a prevalência foi maior (2,8%) quando considerado VDRL positivo, independente do título. Análise multivariada mostrou associação significativa da sífilis com comportamento homo ou bissexual [OR=6,80; IC95% 1,00­46,20], história prévia de infecções sexualmente transmissíveis [OR=16,30; IC95% 3,61­73,41], tatuagem [OR=6,21; IC95% 1,49­25,84] e uso de cocaína [OR=6,80; IC95% 1,15­40,30]. A prevalência de teste treponêmico positivo foi de 10,4%. Conclusão: A soroprevalência de sífilis ativa nesta população foi semelhante à observada em outros estudos populacionais no Brasil. A alta prevalência nos testes treponêmicos positivos pode ser devida a cicatriz sorológica de infecção curada, mas pode também estar associada a casos de sífilis primária ou tardia, que não foram detectados pelo teste de VDRL.

Brazilian Protocol for Sexually Transmitted Infections 2020: syphilis diagnostic tests.

The recommendations for diagnostic tests for investigating syphilis are part of the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections and the Technical Manual for Syphilis Diagnosis, published by the Brazilian Ministry of Health. These recommendations were developed based on scientific evidence and discussions with a panel of experts. This article presents direct tests to detect Treponema pallidum in lesions and algorithms that combine treponemal and non-treponemal antibody tests to assist in syphilis diagnosis, with the aim of contributing to the efforts of health service managers and health professionals in qualifying health care. The article also covers the use of non-treponemal tests to investigate neurosyphilis and guidelines for interpreting non-treponemal antibody titers in monitoring the treatment and diagnosis of congenital syphilis, as well as prospects for innovations in diagnosis. The critical role of rapid immunochromatographic treponemal tests for public health and for addressing syphilis is also highlighted. Highlighted excerpt: During the natural evolution of syphilis, activity periods with distinct clinical, immunological, and histopathological characteristics are interspersed with latent periods when there are no signs or symptoms, making access to tests critical for early diagnosis.

[Brazilian Protocol for Sexually Transmitted Infections 2020: syphilis diagnostic tests]. / Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: testes diagnósticos para sífilis.

The recommendations for diagnostic tests for investigating syphilis are part of the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections and the Technical Manual for Syphilis Diagnosis, published by the Brazilian Ministry of Health. These recommendations were developed based on scientific evidence and discussions with a panel of experts. Aiming to contribute to the efforts of health service managers and health professionals in qualifying health care, this article presents the use of direct tests to detect Treponema pallidum in lesions, as well as algorithms that combine treponemal and non-treponemal antibody tests to assist in the diagnosis of syphilis. The article also covers use of non-treponemal tests to investigate neurosyphilis and guidelines for interpreting non-treponemal antibody titers in monitoring treatment and diagnosis of congenital syphilis, as well as prospects for innovations in diagnosis. The important role of rapid immunochromatographic treponemal tests for public health and for addressing syphilis is also highlighted.

As recomendações de testes diagnósticos para investigação da sífilis compõem o Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis e o Manual Técnico para Diagnóstico de Sífilis, publicados pelo Ministério da Saúde do Brasil. Tais recomendações foram elaboradas com base em evidências científicas e discussões com painel de especialistas. Visando contribuir com gestores e profissionais de saúde na qualificação da assistência, este artigo apresenta o uso dos exames diretos para detecção de Treponema pallidum em lesões, assim como algoritmos que combinam testes imunológicos treponêmicos e não treponêmicos para auxiliar no diagnóstico da sífilis. O artigo também apresenta o uso dos testes não treponêmicos para investigação de neurossífilis e orientações para interpretação do título dos anticorpos não treponêmicos no monitoramento do tratamento e diagnóstico de sífilis congênita, bem como as perspectivas futuras de inovações em diagnóstico. Ressalta-se, além disso, o importante papel dos testes rápidos imunocromatográficos treponêmicos para a saúde pública e o enfrentamento da sífilis.

Las recomendaciones de las pruebas de diagnóstico de sífilis forman parte del Protocolo Clínico y Directrices Terapéuticas para la Atención Integral a las Personas con Infecciones de Transmisión Sexual y del Manual Técnico para el Diagnóstico de Sífilis, publicados por el Ministerio de Salud de Brasil. Estos documentos fueron preparados en base a la evidencia científica y discusiones con especialistas. Con el objetivo de contribuir con los administradores y profesionales de la salud pública en la calificación de la atención, este artículo presenta el uso de pruebas directas para la detección de Treponema pallidum en lesiones, además de algoritmos que combinan pruebas treponémicas y no treponémicas para ayudar al diagnóstico. También se presentan las pruebas no treponémicas para investigar la neurosífilis y orientaciones para la interpretación de títulos de anticuerpos no treponémicos para monitorear la respuesta al tratamiento y diagnosticar la sífilis congénita, así como las perspectivas futuras para las innovaciones diagnósticas. También se enfatiza el uso de pruebas inmunocromatográficas treponémicas rápidas como una herramienta importante para la salud pública y para el enfrentamiento de la sífilis.

Follow-up and outcome of the twelve-year experience in adult patients with acute promyelocytic leukemia

ABSTRACT Acute promyelocytic leukemia is a subtype of acute myeloid leukemia, characterized by the presence of neoplastic promyelocytes, due to the reciprocal balanced translocation between chromosomes 15 and 17. Currently, with the use of agents that act directly on this molecular change, such as all-trans retinoic acid and arsenic trioxide, APL has shifted from a highly mortal to a curable disease. However, some cases are still at high risk of death, especially early death, and acquiring a better understanding of the clinical and biological factors involving APL is needed to correctly identify and treat such cases. The early suspected diagnosis and prompt initiation of the target therapy are important for better response rates. The follow-up and outcomes, using real-life data from 44 consecutive APL patients, were studied between 2001 and 2013. The overall survival rate was 82.7% and early death was 16%. Almost all patient deaths were due to severe bleeding, which was confirmed by multivariate analysis, as the most important prognostic factor leading to death. A better understanding the pathogenesis of the hemorrhagic complications in APL is needed, as well as the risk factors associated with early death in APL patients, as this has become synonymous with overall mortality.

Follow-up and outcome of the twelve-year experience in adult patients with acute promyelocytic leukemia.

Acute promyelocytic leukemia is a subtype of acute myeloid leukemia, characterized by the presence of neoplastic promyelocytes, due to the reciprocal balanced translocation between chromosomes 15 and 17. Currently, with the use of agents that act directly on this molecular change, such as all-trans retinoic acid and arsenic trioxide, APL has shifted from a highly mortal to a curable disease. However, some cases are still at high risk of death, especially early death, and acquiring a better understanding of the clinical and biological factors involving APL is needed to correctly identify and treat such cases. The early suspected diagnosis and prompt initiation of the target therapy are important for better response rates. The follow-up and outcomes, using real-life data from 44 consecutive APL patients, were studied between 2001 and 2013. The overall survival rate was 82.7% and early death was 16%. Almost all patient deaths were due to severe bleeding, which was confirmed by multivariate analysis, as the most important prognostic factor leading to death. A better understanding the pathogenesis of the hemorrhagic complications in APL is needed, as well as the risk factors associated with early death in APL patients, as this has become synonymous with overall mortality.

Brazilian protocol for sexually transmitted infections 2020: syphilis diagnostic tests

Abstract The recommendations for diagnostic tests for investigating syphilis are part of the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections and the Technical Manual for Syphilis Diagnosis, published by the Brazilian Ministry of Health. These recommendations were developed based on scientific evidence and discussions with a panel of experts. This article presents direct tests to detect Treponema pallidum in lesions and algorithms that combine treponemal and non-treponemal antibody tests to assist in syphilis diagnosis, with the aim of contributing to the efforts of health service managers and health professionals in qualifying health care. The article also covers the use of non-treponemal tests to investigate neurosyphilis and guidelines for interpreting non-treponemal antibody titers in monitoring the treatment and diagnosis of congenital syphilis, as well as prospects for innovations in diagnosis. The critical role of rapid immunochromatographic treponemal tests for public health and for addressing syphilis is also highlighted.

Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: testes diagnósticos para sífilis / Brazilian Protocol for Sexually Transmitted Infections 2020: syphilis diagnostic tests / Protocolo Brasileño para Infecciones de Transmisión Sexual 2020: pruebas de diagnóstico de sífilis

As recomendações de testes diagnósticos para investigação da sífilis compõem o Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis e o Manual Técnico para Diagnóstico de Sífilis, publicados pelo Ministério da Saúde do Brasil. Tais recomendações foram elaboradas com base em evidências científicas e discussões com painel de especialistas. Visando contribuir com gestores e profissionais de saúde na qualificação da assistência, este artigo apresenta o uso dos exames diretos para detecção de Treponema pallidum em lesões, assim como algoritmos que combinam testes imunológicos treponêmicos e não treponêmicos para auxiliar no diagnóstico da sífilis. O artigo também apresenta o uso dos testes não treponêmicos para investigação de neurossífilis e orientações para interpretação do título dos anticorpos não treponêmicos no monitoramento do tratamento e diagnóstico de sífilis congênita, bem como as perspectivas futuras de inovações em diagnóstico. Ressalta-se, além disso, o importante papel dos testes rápidos imunocromatográficos treponêmicos para a saúde pública e o enfrentamento da sífilis.

The recommendations for diagnostic tests for investigating syphilis are part of the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections and the Technical Manual for Syphilis Diagnosis, published by the Brazilian Ministry of Health. These recommendations were developed based on scientific evidence and discussions with a panel of experts. Aiming to contribute to the efforts of health service managers and health professionals in qualifying health care, this article presents the use of direct tests to detect Treponema pallidum in lesions, as well as algorithms that combine treponemal and non-treponemal antibody tests to assist in the diagnosis of syphilis. The article also covers use of non-treponemal tests to investigate neurosyphilis and guidelines for interpreting non-treponemal antibody titers in monitoring treatment and diagnosis of congenital syphilis, as well as prospects for innovations in diagnosis. The important role of rapid immunochromatographic treponemal tests for public health and for addressing syphilis is also highlighted.

Las recomendaciones de las pruebas de diagnóstico de sífilis forman parte del Protocolo Clínico y Directrices Terapéuticas para la Atención Integral a las Personas con Infecciones de Transmisión Sexual y del Manual Técnico para el Diagnóstico de Sífilis, publicados por el Ministerio de Salud de Brasil. Estos documentos fueron preparados en base a la evidencia científica y discusiones con especialistas. Con el objetivo de contribuir con los administradores y profesionales de la salud pública en la calificación de la atención, este artículo presenta el uso de pruebas directas para la detección de Treponema pallidum en lesiones, además de algoritmos que combinan pruebas treponémicas y no treponémicas para ayudar al diagnóstico. También se presentan las pruebas no treponémicas para investigar la neurosífilis y orientaciones para la interpretación de títulos de anticuerpos no treponémicos para monitorear la respuesta al tratamiento y diagnosticar la sífilis congénita, así como las perspectivas futuras para las innovaciones diagnósticas. También se enfatiza el uso de pruebas inmunocromatográficas treponémicas rápidas como una herramienta importante para la salud pública y para el enfrentamiento de la sífilis.

Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: testes diagnósticos para sífilis / Brazilian Protocol for Sexually Transmitted Infections 2020: syphilis diagnostic tests / Protocolo Brasileño para Infecciones de Transmisión Sexual 2020: pruebas de diagnóstico de sífilis

Resumo As recomendações de testes diagnósticos para investigação da sífilis compõem o Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis e o Manual Técnico para Diagnóstico de Sífilis, publicados pelo Ministério da Saúde do Brasil. Tais recomendações foram elaboradas com base em evidências científicas e discussões com painel de especialistas. Visando contribuir com gestores e profissionais de saúde na qualificação da assistência, este artigo apresenta o uso dos exames diretos para detecção de Treponema pallidum em lesões, assim como algoritmos que combinam testes imunológicos treponêmicos e não treponêmicos para auxiliar no diagnóstico da sífilis. O artigo também apresenta o uso dos testes não treponêmicos para investigação de neurossífilis e orientações para interpretação do título dos anticorpos não treponêmicos no monitoramento do tratamento e diagnóstico de sífilis congênita, bem como as perspectivas futuras de inovações em diagnóstico. Ressalta-se, além disso, o importante papel dos testes rápidos imunocromatográficos treponêmicos para a saúde pública e o enfrentamento da sífilis.

Abstract The recommendations for diagnostic tests for investigating syphilis are part of the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections and the Technical Manual for Syphilis Diagnosis, published by the Brazilian Ministry of Health. These recommendations were developed based on scientific evidence and discussions with a panel of experts. Aiming to contribute to the efforts of health service managers and health professionals in qualifying health care, this article presents the use of direct tests to detect Treponema pallidum in lesions, as well as algorithms that combine treponemal and non-treponemal antibody tests to assist in the diagnosis of syphilis. The article also covers use of non-treponemal tests to investigate neurosyphilis and guidelines for interpreting non-treponemal antibody titers in monitoring treatment and diagnosis of congenital syphilis, as well as prospects for innovations in diagnosis. The important role of rapid immunochromatographic treponemal tests for public health and for addressing syphilis is also highlighted.

Resumen Las recomendaciones de las pruebas de diagnóstico de sífilis forman parte del Protocolo Clínico y Directrices Terapéuticas para la Atención Integral a las Personas con Infecciones de Transmisión Sexual y del Manual Técnico para el Diagnóstico de Sífilis, publicados por el Ministerio de Salud de Brasil. Estos documentos fueron preparados en base a la evidencia científica y discusiones con especialistas. Con el objetivo de contribuir con los administradores y profesionales de la salud pública en la calificación de la atención, este artículo presenta el uso de pruebas directas para la detección de Treponema pallidum en lesiones, además de algoritmos que combinan pruebas treponémicas y no treponémicas para ayudar al diagnóstico. También se presentan las pruebas no treponémicas para investigar la neurosífilis y orientaciones para la interpretación de títulos de anticuerpos no treponémicos para monitorear la respuesta al tratamiento y diagnosticar la sífilis congénita, así como las perspectivas futuras para las innovaciones diagnósticas. También se enfatiza el uso de pruebas inmunocromatográficas treponémicas rápidas como una herramienta importante para la salud pública y para el enfrentamiento de la sífilis.

Cytotoxic mechanisms of primin, a natural quinone isolated from Eugenia hiemalis, on hematological cancer cell lines.

Considering the high morbidity and mortality rates associated with hematological malignancies and the frequent development of drug resistance by these diseases, the search for new cytotoxic agents is an urgent necessity. The new compounds should present higher efficiency and specificity in inducing tumor cell death, be easily administered and have little or negligible adverse effects. Quinones have been reported in the literature by their several pharmacological properties, including antitumor activity, thus, the aim of this study was to investigate the cytotoxic effect of primin, a natural quinone, on hematological malignancies cell lines. Primin was highly cytotoxic against the three cell lines included in this study (K562, Jurkat and MM.1S) in a concentration- and time-dependent manner, as demonstrated by the MTT method. The compound triggered an apoptotic-like cell death, as observed by ethidium bromide/acridine orange staining, DNA fragmentation and phosphatidylserine exposure after labeling with Annexin V. Both intrinsic and extrinsic apoptosis are involved in cell death induced by primin, as well as the modulation of cell proliferation marker KI-67. The activation of intrinsic apoptosis appears to be related to a decreased Bcl-2 expression and increased Bax expression. While the increase in FasR expression signals activate extrinsic apoptosis. The results suggest that primin is a promising natural molecule that could be used in hematological malignancies therapy or as prototypes for the development of new chemotherapics.

Synthesis of novel pyrazoline derivatives and the evaluation of death mechanisms involved in their antileukemic activity.

Malignant neoplasms are one of the leading causes of death worldwide and hematologic malignancies, including acute leukemia (AL) is one of the most relevant cancer types. Current available chemotherapeutics are associated with high morbidity and mortality rates, therefore, the search for new molecules with antitumor activity, specific and selective for neoplastic cells, became a great challenge for researchers in the oncology field. As pyrazolines stand out in the literature for their great variety of biological activities, the aim of this study was to synthesize and evaluate the antileukemic activity of five new pyrazoline derivatives. All pyrazolines showed adequate physicochemical properties for a good oral bioavailability. The two unpublished and most effective pyrazoline derivatives have been selected for further experiments. These compounds are highly selective for leukemic cells when compared to non-neoplastic cells and did not cause lysis on human red blood cells. Additionally, selected pyrazolines induced cell cycle arrest at G0/G1 phase and decreased cell proliferation marker KI67. Apoptotic cell death induced by selected pyrazolines was confirmed by morphological analysis, assessment of phosphatidylserine residue exposure and DNA fragmentation. Several factors indicate that both intrinsic and extrinsic apoptosis occurred. These were: increased FasR expression; the predominance of Bax in relation to Bcl-2; the loss of mitochondrial membrane potential; AIF release; decreased expression of survivin (an antiapoptotic protein); and the activation of caspase-3. The selected pyrazolines were also found to be cytotoxic against neoplastic cells collected from the peripheral blood and bone marrow of patients with different subtypes of acute leukemia.

MICONIDINE acetate, a new selective and cytotoxic compound with synergic potential, induces cell cycle arrest and apoptosis in leukemia cells.

Plants are important sources of biologically active compounds and they provide unlimited opportunities for the discovery and development of new drug leads, including new chemotherapeutics. Miconidin acetate (MA) is a hydroquinone derivative isolated from E. hiemalis. In this study we demonstrated that MA was cytotoxic against acute leukemia (AL), solid tumor cells and cancer stem cells, with the strongest effect exhibited against AL. Furthermore, it was non-cytotoxic against non-tumor cells and did not cause significant hemolysis. MA blocks the G2/M phase and causes cytostatic effects, acting in a similar way to dexamethasone by increasing PML expression. The compound also triggered intrinsic and extrinsic apoptosis by modulating Bax, FasR and survivin expression. This led to an extensive mitochondrial damage that resulted in AIF, cytochrome c and endonuclease G release, caspase-3 and PARP cleavage and DNA fragmentation. We have further demonstrated that MA was strongly cytotoxic against neoplastic cells collected from patients with different AL subtypes. Interestingly, MA increased the cytotoxic effect of chemotherapeutics cytarabine and vincristine. This study indicates that MA may be a new agent for AL and highlights its potential as a new source of anticancer drugs. Graphical abstract MA blocks G2/M phase with PML expression and KI67 inhibition, ROS generation and intrinsic and extrinsic apoptosis, leading to mitochondrial damage, caspase 3 and PARP cleavage and DNA fragmentation.

Evaluation of seven rapid tests for syphilis available in Brazil using defibrinated plasma panels.

OBJECTIVES: In 2012, the WHO estimated that 6 million new cases of syphilis per year would occur worldwide, including 937 000 in Brazil. Early diagnosis and treatment of syphilis are essential to reduce morbidity and prevent transmission. The availability of rapid tests (RTs) for this diagnosis means that testing can be performed more quickly, as a point-of-care test, even in non-laboratory environments and requires only simple technical training to antibodies detection. The objective of this study was to evaluate the performance and operational aspects of seven commercially available RTs for syphilis in Brazil. METHODS: Seven rapid treponemal tests were evaluated for sensitivity, specificity, accuracy and Kappa value, according to a panel composed of 493 members. The operational performance of the assay was also determined for these tests. RESULTS: The seven RTs showed sensitivity ranging from 94.5% to 100% when compared with the reference tests and specificity of between 91.5% and 100%. All the RTs evaluated presented good operational performance, and only one failed to present the minimum specificity as defined by Brazil's Ministry of Health. CONCLUSION: All the tests presented good operational performance, and the professionals who performed them considered them to be easy to use and interpret. This evaluation is important for making informed choices of tests to be used in the Brazilian Unified Health System.

Cytotoxic effect of a novel naphthylchalcone against multiple cancer cells focusing on hematologic malignancies.

Chalcones are natural compounds described in the literature by its several properties including cytotoxic activity against several tumor types. Considering that the search for new chemotherapeutic agents is still necessary, the aim of this study was to investigate the cytotoxic mechanisms involved in cell death induced by a synthetic chalcone (A23) on different tumor cells. Chalcone A23 reduced the cell viability of twelve tumor cell lines in a concentration and time dependent manner and it was more cytotoxic against acute leukemia cells. Interestingly, the compound was non cytotoxic to normal cells and non-hemolytic to normal red blood cells. Chalcone A23 decreased the expression of cell proliferation marker KI-67 and blocked the G2/M phase in both K562 and Jurkat cell lines. Cells treated with A23 showed morphological features suggestive of apoptosis, the "latter pattern" in agarose gel, the externalization of phosphatidylserine and caspase-3 and PARP cleavage. Chalcone A23 significantly reduced the mitochondrial membrane potential, decreased the expression of anti-apoptotic proteins Bcl-2 and survivin and increased the expression of pro-apoptotic protein Bax, confirming the involvement of the intrinsic pathway. The increased mitochondrial permeability resulted in the release of AIF, cytochrome c and endonuclease G from the mitochondria to the cytosol. In addition, chalcone A23 increased the expression of FasR and induced Bid cleavage, showing the involvement of the extrinsic pathway. Finally, chalcone A23 seems to have a synergic effect with the chemotherapy drugs cytarabine and vincristine. These results suggest that A23 is an interesting compound with strong and selective anti-tumor activity.

Effects of 1,3,5-triphenyl-4,5-dihydro-1H-pyrazole derivatives on cell-cycle and apoptosis in human acute leukemia cell lines.

Pyrazoline is an important 5-membered nitrogen heterocycle that has been extensively researched. Ten derivatives were synthesized and tested for antileukemic effects on 2 human acute leukemia cell lines, K562 and Jurkat. The most cytotoxic of these derivatives, compound 21, was chosen for investigation of cytotoxicity mechanisms. The results obtained with selectivity calculations revealed that compound 21 is more selective for acute leukemia (K562 and Jurkat cell lines) than for other tumor cell lines. Moreover, compound 21 was not cytotoxic to normal cell lines, indicating a potential use in clinical tests. Compound 21 caused a significant cell cycle arrest in the S-phase in Jurkat cells and increased the proportion of cells in the sub G0/G1 phase in both cell lines. Cells treated with compound 21 demonstrated morphological changes characteristic of apoptosis in the EB/AO assay, confirmed by externalization of phosphatidylserine by the annexin V - fluorescein isothiocyanate method and by DNA fragmentation. An investigation of cytotoxicity mechanisms suggests the involvement of an intrinsic apoptosis pathway due to mitochondrial damage and an increase in the ratio of mitochondrial Bax/Bcl2. Pyrazoline 21 obeyed Lipinski's "rule of five" for drug-likeness. Based on these preliminary results, the antileukemic activity of compound 21 makes it a potential anticancer agent.