Abstract Since the inception of COVID-19 pandemic, there has been a challenging race for the development of precise diagnostic tests. Specific SARS-CoV-2 serological assays are the main tools used to estimate the rate of past infections or herd immunity in epidemiological studies, in addition to being helpful in guiding public health management policies. In this study, an in-house ELISA based on the construct of SARS-CoV-2 nucleocapsid (N) proteins, named rCoV2, rCoV4, and rCoV7, showed diagnostic performance for the detection of IgG antibodies. Sensitivity was evaluated in serum samples from patients with mild to moderate or severe COVID-19 infections, which were collected at different time points, while specificity was evaluated using pre-pandemic sera. In samples from mild to moderate cases obtained ≥16 days after the onset of symptoms, the sensitivities for rCoV2, rCoV4, and rCoV7 were 66.7%, 75%, and 77.8%, respectively. For samples from severe cases, the sensitivity was above 80% for all constructs. All proteins showed high specificity (94-98%). Overall, rCoV7 (C-terminus N-protein portion) showed better diagnostic performance, with 62.3% sensitivity in moderate and severe cases and 96.6% specificity. The SARS-CoV-2 ELISA using N-protein-based constructs could be a promisor tool for investigate the epidemiology of COVID-19 and monitor population-level serosurveillance. COVID19 COVID 19 COVID-1 pandemic tests SARSCoV2 SARSCoV SARS CoV 2 SARS-CoV- studies policies study inhouse house N (N rCoV2 rCoV rCoV4 antibodies points prepandemic pre sera 16 ≥1 symptoms 667 66 7 66.7% 75 75% 778 77 8 77.8% respectively 80 9498%. 9498 94 98% . 98 (94-98%) Overall Cterminus C terminus Nprotein protein portion 623 62 3 62.3 966 96 6 96.6 Nproteinbased populationlevel population level serosurveillance COVID1 1 COVID- SARS-CoV ≥ 66.7 77.8 9498% 949 9 (94-98% 62. 96. 66. 77. (94-98 (94-9 (94- (94 (9 (

ABSTRACT Differential diagnosis of coronavirus disease 2019 (COVID-19) from other febrile diseases is one of several challenges imposed by the pandemic. We present a case of severe malaria and COVID-19 coinfection in a non-malaria-endemic region. A 44-year-old female with malaise, fever, hypotension, jaundice, and enlarged liver and spleen was admitted to the intensive care unit. Reverse transcription-quantitative PCR results for severe acute respiratory syndrome coronavirus 2 were positive. Rapid tests, microscopy, and quantitative PCR were positive for Plasmodium vivax. Cytokine storm profiles were identified. We could not determine whether the severe vivax malaria in our patient was triggered by COVID-19 coinfection. 201 COVID19 COVID 19 (COVID-19 pandemic COVID-1 nonmalariaendemic non endemic region 44yearold yearold 44 year old malaise fever hypotension jaundice unit transcriptionquantitative transcription tests microscopy identified 20 COVID1 1 (COVID-1 COVID- 4 (COVID- (COVID

ABSTRACT There is a consensus that the antifungal repertoire for the treatment of cryptococcal infections is limited. Standard treatment involves the administration of an antifungal drug derived from natural sources (i.e., amphotericin B) and two other drugs developed synthetically (i.e., flucytosine and fluconazole). Despite treatment, the mortality rates associated with fungal cryptococcosis are high. Amphotericin B and flucytosine are toxic, require intravenous administration, and are usually unavailable in low-income countries because of their high cost. However, fluconazole is cost-effective, widely available, and harmless with regard to its side effects. However, fluconazole is a fungistatic agent that has contributed considerably to the increase in fungal resistance and frequent relapses in patients with cryptococcal meningitis. Therefore, there is an unquestionable need to identify new alternatives or adjuvants to conventional drugs for the treatment of cryptococcosis. A potential antifungal agent should be able to kill cryptococci and “bypass” the virulence mechanism of the yeast. Furthermore, it should have fungicidal action, low toxicity, high selectivity, easily penetrate the central nervous system, and widely available. In this review, we describe cryptococcosis, its conventional therapy, and failures arising from the use of drugs traditionally considered to be the reference standard. Additionally, we present the approaches used for the discovery of new drugs to counteract cryptococcosis, ranging from the conventional screening of natural products to the inclusion of structural modifications to optimize anticryptococcal activity, as well as drug repositioning and combined therapies. limited i.e., ie i e (i.e. fluconazole. . fluconazole) toxic lowincome income cost However costeffective, costeffective effective, effective cost-effective available effects meningitis Therefore bypass “bypass yeast Furthermore action toxicity selectivity system review therapy standard Additionally activity therapies i.e. (i.e i.e

ABSTRACT Background: Women living with human immunodeficiency virus (HIV) (WLWH) are more likely to be infected with the oncogenic human papillomavirus (HPV). We assessed the prevalence of high-risk (HR) (16/18/31/33/35/39/45/51/52/56/58/59/68/73/82), probable high-risk (pHR) (26/53/66), and low-risk (LR) (6/11/40/42/43/44/54/61/70) HPV types and their associated risk factors. Methods: This cross-sectional study of WLWH aged 18-64 years included one laboratory and eight HIV-specialty healthcare facilities in the pilot network. Descriptive statistics were used to assess sociodemographic and behavioral characteristics. Adjusted analyses were conducted to evaluate risk factors associated with HR and/or pHR HPV infection in WLWH. Results: From May/2021 to May/2022, 1,914 (92.5%) WLWH participated in the pilot study and had valid HPV-DNA results of self-collected vaginal samples. The median age of the participants was 45 years, 60.1% had ≥ 9 years of schooling, 80.5% were ≤ 18 years at first sexual intercourse, and 51.7% had > 4 sexual partners throughout life. The prevalence of any HPV type, HR HPV, pHR HPV, and LR HPV was 65.8%, 49.6%, 16.7%, and 40.0%, respectively. Age was inversely associated with pHR and/or HR-HPV (p < 0.001), and education level was inversely associated with HR-HPV (p = 0.003) types. Any HR or pHR was associated with being single (p = 0.029) and exchanging sex for drugs (p = 0.037). Conclusions: The prevalence of HPV, especially HR HPV, among WLWH is high in Brazil, highlighting the need for HPV screening in this population. Self-collection of vaginal samples is an important strategy for increasing testing access. Background HIV (HIV (WLWH HPV. . (HPV) highrisk (HR 16/18/31/33/35/39/45/51/52/56/58/59/68/73/82, 161831333539455152565859687382 16/18/31/33/35/39/45/51/52/56/58/59/68/73/82 , 16 31 33 35 39 51 52 56 58 59 68 73 82 (16/18/31/33/35/39/45/51/52/56/58/59/68/73/82) (pHR 26/53/66, 265366 26/53/66 26 53 66 (26/53/66) lowrisk low (LR 6/11/40/42/43/44/54/61/70 61140424344546170 6 11 40 42 43 44 54 61 70 (6/11/40/42/43/44/54/61/70 Methods crosssectional cross sectional 1864 64 18-6 HIVspecialty specialty network characteristics andor Results May2021 May 2021 May/202 May2022 2022 May/2022 1914 1 914 1,91 92.5% 925 92 5 (92.5% HPVDNA DNA selfcollected self collected 601 60 60.1 schooling 805 80 80.5 intercourse 517 7 51.7 life type 658 65 8 65.8% 496 49 49.6% 167 16.7% 400 0 40.0% respectively HRHPV p 0.001, 0001 0.001 001 0.001) 0.003 0003 003 0.029 0029 029 0.037. 0037 0.037 037 0.037) Conclusions Brazil population Selfcollection Self collection access (HPV 16183133353945515256585968738 16/18/31/33/35/39/45/51/52/56/58/59/68/73/8 3 (16/18/31/33/35/39/45/51/52/56/58/59/68/73/82 26536 26/53/6 2 (26/53/66 6/11/40/42/43/44/54/61/7 6114042434454617 (6/11/40/42/43/44/54/61/7 186 18- May202 202 May/20 191 91 1,9 92.5 (92.5 60. 80. 51. 65.8 49.6 16.7 40.0 000 0.00 00 0.02 002 02 0.03 03 1618313335394551525658596873 16/18/31/33/35/39/45/51/52/56/58/59/68/73/ (16/18/31/33/35/39/45/51/52/56/58/59/68/73/8 2653 26/53/ (26/53/6 6/11/40/42/43/44/54/61/ 611404243445461 (6/11/40/42/43/44/54/61/ May20 20 May/2 19 1, 92. 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Abstract Genetic variability in the host metabolism of antimalarial drugs influenced by the polymorphisms of cytochrome P450 (CYP) could lead to significant changes in antimalarial treatment response. However, little is known about the frequency of alleles CYP2B6, CYP2C8, and CYP2D6 in an Amazonian population, especially with vivax malaria. Therefore, this study aimed to determine the frequency of CYP alleles CYP2B6*6, CYP2C8*3, and CYP2D6*4 in patients with vivax malaria. The study included 231 patients with vivax malaria treated at a health care reference in Manaus, northern Brazil. A sample of peripheral blood from each subject was collected to perform DNA extraction and genotypic analysis. Genotyping of polymorphisms was performed by allelic discrimination using Real-time polymerase chain reaction. The CYP2D6*4 allele was the most prevalent among patients who developed severe malaria. The frequencies of the CYP2B6*6 and CYP2D6*4 were not different between the severe and uncomplicated malaria. There was a significant association between heterozygous CYP2D6*4 and severe cases of malaria. The results are in agreement with other reports described in the literature for different populations. Future studies are needed to understand the clinical implications of the polymorphisms in patients with vivax malaria. P P45 (CYP response However CYP2B6 CYPB B CYP2C8 CYPC C CYPD D CYP2D population Therefore CYP2B66 6 CYP2C83 3 CYP2C8*3 CYP2D64 4 CYP2D6* 23 Manaus Brazil analysis Realtime Real time reaction CYP2B6* populations P4 CYP2B CYP2C CYP2C8* 2

ABSTRACT Background: Although primaquine (PQ) is indicated for G6PD-deficient patients, data on weekly PQ use in Brazil are limited. Methods: We aimed to investigate malaria recurrences among participants receiving daily and weekly PQ treatments in a real-life setting of two municipalities in the Amazon between 2019 and 2020. Results: Patients receiving weekly PQ treatment had a lower risk of recurrence than those receiving daily PQ treatment (risk ratio: 0.62, 95% confidence interval: 0.41-0.94), using a model adjusted for study site. Conclusions: Weekly PQ use did not increase the risk of malaria recurrence. Further studies with larger populations are warranted.

Resumo Objetivo: descrever os arranjos tecnoassistenciais desenvolvidos no âmbito da gestão do trabalho na rede de atenção à pandemia de COVID-19, na perspectiva de gestores. Método: pesquisa qualitativa, do tipo caso único incorporado, com 23 gestores de uma Rede de Atenção à Saúde. Análise aplicada em dois ciclos de codificação temática, com o auxílio do software ATLAS.ti. Resultados: os arranjos foram analisados em categorias vinculadas à: atenção à saúde; gestão; incorporação de tecnologias; implantação de hospital de campanha; e análise retrospectiva das experiências como um todo. Houve destaque para a implantação de fluxo de atendimentos, boletins de saúde virtuais, telemonitoramento, chatbots, uso de aplicativos, implantação de hospitais de campanha e da urgência básica no âmbito de Unidades Básicas de Saúde. Identificou-se a hiperjudicialização no sistema; fragilidades na gestão das informações, intersetorialidade e condução técnico-política em âmbito nacional; o protagonismo dos enfermeiros em cargos de gestão e para o enfrentamento da pandemia. Conclusão: apesar do despreparo dos serviços de saúde para o enfrentamento da pandemia, a resiliência dos atores promoveu dinamicidade e arranjos tecnoassistenciais no âmbito da gestão e do cuidado humanizado. O estudo tem potencial contribuição para qualificação das práticas de gestão e desenvolvimento de políticas públicas.

Abstract Objective: to describe the technical-assistance arrangements developed within the scope of work management in the COVID-19 pandemic care network, from the managers’ perspective. Method: a qualitative research study, of the incorporated single case type, conducted with 23 managers of a Health Care Network. The analysis was applied in two thematic coding cycles, with the aid of the ATLAS.ti software. Results: the arrangements were analyzed in categories related to health care; management; incorporation of technologies; implementation of a field hospital; and retrospective analysis of the experiences as a whole. There was emphasis on the implementation of care flows, virtual health bulletins, Telemonitoring, chatbots, use of applications, and implementation of field hospitals and of basic urgency services within the scope of the Basic Health Units. Hyperjudicialization in the system was identified; as well as weaknesses in information management, intersectoriality and technical-political leadership at the national level; the role of nurses in management positions and for coping with the pandemic. Conclusion: despite the health services’ unpreparedness to face the pandemic, the actors’ resilience promoted dynamism and technical-assistance arrangements in the context of management and humanized care. The study has a potential to contribute to the qualification of the public policy management and development practices.

Resumen Objetivo: describir los preparativos tecnoasistenciales que se desarrollaron en el ámbito de la gestión del trabajo en la red de atención de la pandemia de COVID-19, desde la perspectiva de los gestores. Método: investigación cualitativa, del tipo caso único incorporado, con 23 gestores de una Red de Atención de Salud. Análisis aplicado en dos ciclos de codificación temática, con ayuda del software ATLAS.ti. Resultados: los preparativos fueron analizados en categorías relacionadas con: la atención de la salud; la administración; la incorporación de tecnologías; la implementación de un hospital de campaña; y el análisis retrospectivo de las experiencias en general. Se destacaron la implementación del flujo de atención, los boletines virtuales de salud, el telemonitoreo, los chatbots, el uso de aplicaciones, la implementación de hospitales de campaña y emergencias básicas en el ámbito de las Unidades Básicas de Salud. Se identificaron la hiperjudicialización en el sistema; las debilidades en la gestión de la información, la intersectorialidad y conducción técnico-política a nivel nacional; el protagonismo de los enfermeros en cargos de gestión y para hacer frente a la pandemia. Conclusión: a pesar de la falta de preparación de los servicios de salud para enfrentar la pandemia, la resiliencia de los actores promovió el dinamismo y los preparativos tecnoasistenciales en el ámbito de la gestión y de la atención humanizada. El estudio tiene una contribución potencial para la calificación de las prácticas de gestión y el desarrollo de políticas públicas.

BACKGROUND Understanding the epidemiology of malaria through the molecular force of the blood-stage infection of Plasmodium vivax (molFOB) may provide a detailed assessment of malaria transmission. OBJECTIVES In this study, we investigated risk factors and spatial-temporal patterns of incidence of Plasmodium infection and clinical malaria episodes in three peri-urban communities of Manaus, Western Brazilian Amazon. METHODS Monthly samples were collected in a cohort of 1,274 individuals between April 2013 and March 2014. DNA samples were subject to Plasmodium species. molFOB was calculated by counting the number of genotypes observed on each visit, which had not been present in the preceding two visits and adjusting these counts by the respective times-at-risk. FINDINGS Respectively, 77.8% and 97.2% of the population remained free of P. vivax and P. falciparum infection. Expected heterozygosity for P. vivax was 0.69 for MSP1_F3 and 0.86 for MS2. Multiplicity of infection in P. vivax was close to the value of 1. The season was associated with P. vivax positivity [adjusted hazard ratio (aHR) 2.6 (1.9-5.7)] and clinical disease [aHR 10.6 (2.4-47.2)]. P. falciparum infection was associated with previous malarial episodes [HR 9.7 (4.5-20.9)]. Subjects who reported possession of a bed net [incidence rate ratio (IRR) 1.6 (1.2-2.2)] or previous malaria episodes [IRR 3.0 (2.0-4.5)] were found to have significantly higher P. vivax molFOB. MAIN CONCLUSIONS Overall, P. vivax infection prevailed in the area and infections were mostly observed as monoclonal. Previous malaria episodes were associated with significantly higher P. vivax molFOB.

BACKGROUND Plasmodium vivax, the major cause of malaria in Latin America, has a large subtelomeric multigene family called vir. In the P. vivax genome, about 20% of its sequences are vir genes. Vir antigens are grouped in subfamilies according to their sequence similarities and have been shown to have distinct roles and subcellular locations. However, little is known about vir subfamilies, especially when comes to their functions. OBJECTIVE To evaluate the diversity, antigenicity, and adhesiveness of Plasmodium vivax VIR-E. METHODS Vir-E genes were amplified from six P. vivax isolates from Manaus, North of Brazil. The presence of naturally acquired antibodies to recombinant PvBrVIR-E and PvAMA-1 was evaluated by ELISA. Binding capacity of recombinant PvBrVIR-E was assessed by adhesion assay to CHO-ICAM1 cells. FINDINGS Despite vir-E sequence diversity, among those identified sequences, a representative one was chosen to be expressed as recombinant protein. The presence of IgM or IgG antibodies to PvBrVIR-E was detected in 23.75% of the study population while the presence of IgG antibodies to PvAMA-1 antigen was 66.25% in the same population. PvBrVIR-E was adhesive to CHO-ICAM1. MAIN CONCLUSIONS PvBrVIR-E was antigenic and adhesive to CHO-ICAM1.

Abstract Bee venom is a natural toxin composed of several peptides. Massive envenoming causes severe local and systemic reactions. We report two cases of severe bee envenomation, of which one was fatal. We also describe clinical characteristics and immune markers. Both victims suffered from respiratory distress, renal failure, rhabdomyolysis, and shock. They required invasive mechanical ventilation, vasoactive drugs, and renal replacement therapy. Moreover, serum levels of chemokines, cytokines, and cell-free circulating nucleic acids demonstrated an intense inflammatory process. Massive envenoming produced systemic injury in the victims, with an uncontrolled inflammatory response, and a more significant chemotactic response in the fatal case.

BACKGROUND Different strategies for improvement of malaria control and elimination are based on the blockage of malaria parasite transmission to the mosquito vector. These strategies include the drugs that target the plasmodial sexual stages in humans and the early developmental stages inside mosquitoes. OBJECTIVES Here we tested Malaria Box compounds in order to evaluate their activity against male and female gametocytes in Plasmodium berghei, mosquito infection in P. vivax and ookinete formation in both species. METHODS/FINDINGS The membrane feeding assay and the development of ookinetes by a 24 h ex vivo culture and the ookinete yield per 1000 erythrocytes were used to test transmission-blocking potential of the Malaria Box compounds in P. vivax. For P. berghei we used flow cytometry to evaluate male and female gametocyte time course and fluorescence microscopy to check the ookinete development. The two species used in this study showed similar results concerning the compounds’ activity against gametocytes and ookinetes, which were different from those in P. falciparum. In addition, from the eight Malaria Box compounds tested in both species, compounds MMV665830, MMV665878 and MMV665941 were selected as a hit compounds due the high inhibition observed. CONCLUSION Our results showed that P. berghei is suitable as an initial screening system to test compounds against P. vivax.

Abstract INTRODUCTION: Electron microscopy (EM) is a rapid and effective tool that can be used to create images of a whole spectrum of virus-host interactions and, as such, has long been used in the discovery and description of viral mechanisms. METHODS: Electron microscopy was used to evaluate the pulmonary pathologies of postmortem lung sections from three patients who died from infection with SARS-associated coronavirus 2 (SARS-CoV-2), a new member of the Coronaviridae family. RESULTS: Diffuse alveolar damage (DAD) was predominant in all three patients. The early exudative stage was characterized principally by edema and extravasation of red blood cells into the alveolar space with injury to the alveolar epithelial cells; this was followed by detachment, apoptosis, and necrosis of type I and II pneumocytes. The capillaries exhibited congestion, exposure of the basement membrane from denuded endothelial cells, platelet adhesion, fibrin thrombi, and rupture of the capillary walls. The proliferative stage was characterized by pronounced proliferation of type II alveolar pneumocytes and multinucleated giant cells. The cytopathic effect of SARS-CoV-2 was observed both in degenerated type II pneumocytes and freely circulating in the alveoli, with components from virions, macrophages, lymphocytes, and cellular debris. CONCLUSIONS: Viral particles consistent with the characteristics of SARS-CoV-2 were observed mainly in degenerated pneumocytes, in the endothelium, or freely circulating in the alveoli. In the final stage of illness, the alveolar spaces were replaced by fibrosis.

Abstract INTRODUCTION: Snakebites in the Brazilian Amazon are caused mostly by snakes from the Bothrops genus and envenomated patients may suffer from tissue complications. METHODS: This study aimed to identify risk factors for severe tissue complications (STC) in patients with Bothrops snakebite in the Amazonas state, Brazil. RESULTS: Snakebites that were classified as severe and affected female patients with comorbidities presented greater risks of developing STCs. In addition, hospitalizations of patients with STC exceeded 5 days. CONCLUSIONS: Clinical and epidemiological characteristics can prove essential for assessing the evolution of STC and clinical prognosis of patients with Bothrops snakebites.

In the present study, we investigated the genetic diversity of Plasmodium vivax metacaspase 1 (PvMCA1) catalytic domain in two municipalities of the main malaria hotspot in Brazil, i.e., the Juruá Valley, and observed complete sequence identity among all P. vivax field isolates and the Sal-1 reference strain. Analysis of PvMCA1 catalytic domain in different P. vivax genomic sequences publicly available also revealed a high degree of conservation worldwide, with very few amino acid substitutions that were not related to putative histidine and cysteine catalytic residues, whose involvement with the active site of protease was herein predicted by molecular modeling. The genetic conservation presented by PvMCA1 may contribute to its eligibility as a druggable target candidate in vivax malaria.