Influence of water electrolysis on hydrolysis and methanogenesis stages of anaerobic digestion at room temperature: Kinetic and metabolic analysis.

Batch cultures were performed to study hydrolysis and methanogenesis in the presence of an electric field at room temperature (i.e., 23 °C). Kinetic and metabolic analyses using RuO2/Ti electrodes were carried out in short reaction times to avoid biofilm formation, allowing the evaluation of the effect of O2 and H2 produced on anaerobic digestion during the imposition of three electric fields: 1.21, 1.45, and 1.64 V/cm. Results highlighted that at 1.21 V/cm, the electrolysis produced 0.0753 mg O2/L·min, where facultative microorganisms consumed 21 % oxygen, enhancing the hydrolysis phase by 52 %. Additionally, methane production was noticeably improved with an activity of 0.89 ± 0.02 g COD-CH4/g VSS·d, meaning 39 % higher than the control. The imposition of an electric field showed promising results since the methanogenic activity at room temperature was very close to the activities observed in conventional reactors at 35 °C.

Mental health outcomes in frontline healthcare workers in Brazil during the COVID-19 epidemic: Results of an online survey in four regions using respondent-driven sampling (RDS).

BACKGROUND: The COVID-19 pandemic overwhelmed health facilities and presented healthcare workers (HCWs) with a new infectious disease threat. In addition to a sanitary crisis, Brazil still had to face major political, economic, and social challenges. This study aimed to investigate mental health outcomes in frontline HCWs in different regions of the country and at different epidemic times. We also sought to identify the main risk factors associated with these outcomes. METHODS: A cross-sectional online survey using respondent-driven sampling was conducted to recruit physicians (n = 584), nurses (n = 997), and nurse technicians (n = 524) in 4 regions of Brazil (North, Northeast, Southeast, and South) from August 2020 to July 2021. We used standardized instruments to screen for common mental disorders (CMD)(SRQ-20), alcohol misuse (AUDIT-C), depression (PHQ-9), anxiety (GAD-7), and post-traumatic stress disorder (PTSD)(PCL-5). Gile's successive sampling estimator was used to produce weighted estimates. We created a three-cluster data set for each HCW category and developed a hierarchical regression model with three levels: individual characteristics; workplace-related aspects; COVID-19 personal experience. The impact of the epidemic moment on the outcomes was also studied. RESULTS: The prevalence of probable CMD was 26.8-36.9%, alcohol misuse 8.7-13.6%, depression 16.4-21.2%, anxiety 10.8-14.2%, and PTSD 5.9-8.0%. We found a stronger association between mental health outcomes and the following factors: history of psychiatric disorders, female gender, and clinical comorbidities (level 1); work overload and family isolation (level 2); sick leave (level 3). Epidemic variables, such as the number of deaths and trend of deaths by COVID-19, had almost no impact on the outcomes. CONCLUSION: An alarmingly high prevalence of depression and anxiety was found in Brazilian frontline HCWs. Individual factors were the most strongly associated with mental health outcomes. These findings indicate the need to develop programs that provide emotional support, identify professionals at risk and refer them to specialized treatment when necessary.

Valvular Heart Disease Emergencies: A Comprehensive Review Focusing on the Initial Approach in the Emergency Department. / Emergências Relacionadas à Doença Valvar Cardíaca: Uma Revisão Abrangente da Abordagem Inicial no Departamento de Emergência.

Valvular heart disease (VHD) is an increasing health problem worldwide. Patients with VHD may experience several cardiovascular-related emergencies. The management of these patients is a challenge in the emergency department, especially when the previous heart condition is unknown. Specific recommendations for the initial management are currently poor. This integrative review proposes an evidence-based three-step approach from bedside VHD suspicion to the initial treatment of the emergencies. The first step is the suspicion of underlying valvular condition based on signs and symptoms. The second step comprises the attempt to confirm the diagnosis and assessment of VHD severity with complementary tests. Finally, the third step addresses the diagnosis and treatment options for heart failure, atrial fibrillation, valvular thrombosis, acute rheumatic fever, and infective endocarditis. In addition, several images of complementary tests and summary tables are provided for physician support.

A doença valvar cardíaca é um problema de saúde crescente no mundo. Os pacientes com valvopatia podem apresentar diversas emergências cardiovasculares. O manejo desses pacientes é um desafio no departamento de emergência, principalmente quando a condição cardíaca prévia é desconhecida. Atualmente, recomendações específicas para o manejo inicial são limitadas. A presente revisão integrativa propõe uma abordagem baseada em evidência, de três etapas, desde a suspeita de valvopatia à beira do leito até o tratamento inicial das emergências. A primeira etapa é a suspeita de uma condição valvar subjacente com base nos sinais e sintomas. A segunda etapa consiste na tentativa de confirmação diagnóstica e avaliação da gravidade da valvopatia com exames complementares. Finalmente, a terceira etapa aborda as opções diagnósticas e terapêuticas para insuficiência cardíaca, fibrilação atrial, trombose valvar, febre reumática aguda, e endocardite infecciosa. Além disso, apresentamos imagens de exames complementares e tabelas para apoio aos médicos.

Impact of the COVID-19 pandemic on the mental health of frontline healthcare workers in a highly affected region in Brazil.

BACKGROUND: The COVID-19 pandemic had a major impact on the mental health of healthcare workers (HCWs), especially in low and middle-income countries, which had to face additional political, social, and economic challenges. We thus aimed to assess the prevalence of mental health outcomes and the associated factors in HCWs treating COVID-19 patients in one of the most affected regions in Brazil. METHODS: We used the Respondent-Driven Sampling method to assess the risks of COVID-19 infection and symptoms of mental disorders in nurses, nursing technicians, and physicians who worked on the frontline in the metropolitan region of Recife. 865 healthcare workers completed a survey regarding sociodemographic data, work-related risks, and symptoms of mental disorders - SRQ-20 for common mental disorders (CMD); AUDIT-C for problematic alcohol use; GAD-7 for anxiety; PHQ-9 for depression; PCL-5 for post-traumatic stress disorder (PTSD). Gile's successive sampling estimator was used to produce the weighted estimates by professional category. A Poisson regression model with robust variance was used to analyze factors associated with a positive screening for CMD. We will present the results of a cross-sectional analysis of the mental health outcomes after the first peak of COVID-19 - from August 2020 to February 2021. RESULTS: The prevalence ratios for a positive screening for CMD were 34.9% (95% CI: 27.8-41.9) in nurses, 28.6% (95% CI: 21.3-36.0) in physicians, and 26.6% (95% CI: 16.8-36.5) in nursing technicians. Nurses presented a higher prevalence of depressive symptoms (23%). Positive screening for problematic alcohol use (10.5 to14.0%), anxiety (10.4 to 13.3%), and PTSD (3.3 to 4.4%) were similar between the professional categories. The main factors associated with CMD in nurses and physicians were related to an intrinsic susceptibility to mental illness, such as previous or family history of psychiatric disorder, and female sex. Among nurse technicians, work-related factors, such as accidents with biological material, presented the strongest association with CMD. CONCLUSION: The mental health of HCWs fighting COVID-19 in Recife was severely affected. It is crucial that healthcare services provide adequate working conditions and psychological support, investing in programs to promote and protect HCWs mental health.

How does a low-magnitude electric field influence anaerobic digestion in wastewater treatment? A review.

Anaerobic digestion (AD) is a physio-biochemical process widely used for treating industrial or municipal wastewater with concomitant methane production. Several technologies have been tested to improve AD's efficiency, like pretreatments and co-digestion, among others. Recently the imposition of a low-magnitude electric field (LMEF) has been applied at the AD to improve methane yield. Despite the positive results of imputing an electric field, many gaps are not understood yet. Therefore, this review focuses on the biochemical aspects of AD and electric field for a better understanding of the effect of the LMEF on the metabolisms of the AD during wastewater treatment and its application in methane production enhancement.

Comparison of COVID-19 hospitalization costs across care pathways: a patient-level time-driven activity-based costing analysis in a Brazilian hospital.

BACKGROUND: The COVID-19 pandemic raised awareness of the need to better understand where and how patient-level costs are incurred in health care organizations, as health managers and other decision-makers need to plan and quickly adapt to the increasing demand for health care services to meet patients' care needs. Time-driven activity-based costing offers a better understanding of the drivers of cost throughout the care pathway, providing information that can guide decisions on process improvement and resource optimization. This study aims to estimate COVID-19 patient-level hospital costs and to evaluate cost variability considering the in-hospital care pathways of COVID-19 management and the patient clinical classification. METHODS: This is a prospective cohort study that applied time-driven activity-based costing (TDABC) in a Brazilian reference center for COVID-19. Patients hospitalized during the first wave of the disease were selected for their data to be analyzed to estimate in-hospital costs. The cost information was calculated at the patient level and stratified by hospital care pathway and Ordinal Scale for Clinical Improvement (OSCI) category. Multivariable analyses were applied to identify predictors of cost variability in the care pathways that were evaluated. RESULTS: A total of 208 patients were included in the study. Patients followed five different care pathways, of which Emergency + Ward was the most followed (n = 118, 57%). Pathways which included the intensive care unit presented a statistically significant influence on costs per patient (p <  0.001) when compared to Emergency + Ward. The median cost per patient was I$2879 (IQR 1215; 8140) and mean cost per patient was I$6818 (SD 9043). The most expensive care pathway was the ICU only, registering a median cost per patient of I$13,519 (IQR 5637; 23,373) and mean cost per patient of I$17,709 (SD 16,020). All care pathways that included the ICU unit registered a higher cost per patient. CONCLUSIONS: This is one of the first microcosting study for COVID-19 that applied the TDABC methodology and demonstrated how patient-level costs vary as a function of the care pathways followed by patients. These findings can be used to develop value reimbursement strategies that will inform sustainable health policies in middle-income countries such as Brazil.

Emergências Relacionadas à Doença Valvar Cardíaca: Uma Revisão Abrangente da Abordagem Inicial no Departamento de Emergência / Valvular Heart Disease Emergencies: A Comprehensive Review Focusing on the Initial Approach in the Emergency Department

Resumo A doença valvar cardíaca é um problema de saúde crescente no mundo. Os pacientes com valvopatia podem apresentar diversas emergências cardiovasculares. O manejo desses pacientes é um desafio no departamento de emergência, principalmente quando a condição cardíaca prévia é desconhecida. Atualmente, recomendações específicas para o manejo inicial são limitadas. A presente revisão integrativa propõe uma abordagem baseada em evidência, de três etapas, desde a suspeita de valvopatia à beira do leito até o tratamento inicial das emergências. A primeira etapa é a suspeita de uma condição valvar subjacente com base nos sinais e sintomas. A segunda etapa consiste na tentativa de confirmação diagnóstica e avaliação da gravidade da valvopatia com exames complementares. Finalmente, a terceira etapa aborda as opções diagnósticas e terapêuticas para insuficiência cardíaca, fibrilação atrial, trombose valvar, febre reumática aguda, e endocardite infecciosa. Além disso, apresentamos imagens de exames complementares e tabelas para apoio aos médicos.

Abstract Valvular heart disease (VHD) is an increasing health problem worldwide. Patients with VHD may experience several cardiovascular-related emergencies. The management of these patients is a challenge in the emergency department, especially when the previous heart condition is unknown. Specific recommendations for the initial management are currently poor. This integrative review proposes an evidence-based three-step approach from bedside VHD suspicion to the initial treatment of the emergencies. The first step is the suspicion of underlying valvular condition based on signs and symptoms. The second step comprises the attempt to confirm the diagnosis and assessment of VHD severity with complementary tests. Finally, the third step addresses the diagnosis and treatment options for heart failure, atrial fibrillation, valvular thrombosis, acute rheumatic fever, and infective endocarditis. In addition, several images of complementary tests and summary tables are provided for physician support.

The cost of total hip replacement: a multicentric TDABC comparison in Brazil / O custo da artroplastia de quadril: uma comparação multicêntrica TDABC no Brasil

Objective: Monitoring costs is critical in searching for a more effective healthcare system. This study aimed to comprehend the care pathway and measure the costs associated with hip replacement surgeries in different hospitals in Brazil. Methods: The time-driven activity-based costing method was applied for cost data collection and analyses. Data on 62 patients were retrieved from five public hospitals. A descriptive cost analysis was followed by a comprehensive analysis of the variability in each hospital's care process, leading to suggestions for cost-saving opportunities along with the surgical care pathway. As a final analysis, the cost of surgical treatment was contrasted with the national reimbursement fee. Results: The mean cost per patient of the total sample was $5,784 (MIN-MAX $2,525.9-$9,557.8). Pre- and post-surgery hospitalization periods demonstrated the highest variability in length of time and resource consumption among centers. Compared to the national best practice fee, the average cost per inpatient total hip arthroplasty (THA) pathway from all six hospitals was approximately 7x the national reimbursement. Conclusion: The application of the TDABC allowed us to identify differences in the surgical care pathway among hospitals, which could be explored in further studies aimed at designing a benchmark surgical pathway. Differences in how the treatment is delivered to patients also justified the high-cost variability among centers.

Objetivo: O custo do monitoramento é um elemento-chave na busca contínua por um sistema de saúde mais eficaz. O objetivo deste estudo foi compreender a trajetória assistencial e mensurar os custos associados às cirurgias de artroplastia do quadril em diferentes hospitais do Brasil. Métodos: O método de custeio baseado em atividades orientado pelo tempo foi aplicado para a coleta e análise de dados de custos. Os dados de 62 pacientes foram recuperados de cinco hospitais públicos. Uma análise descritiva de custos foi seguida por uma análise abrangente da variabilidade no processo de atendimento de cada hospital, levando a sugestões de oportunidades de redução de custos junto com a via de atendimento cirúrgico. Como análise final, o custo do tratamento cirúrgico foi contrastado com o valor de reembolso nacional. Resultados: O custo médio por paciente da amostra total foi de $ 5.784 (MIN-MAX $ 2.525,9-$ 9.557,8). Os períodos de internação pré e pós-operatórios demonstraram a maior variabilidade no tempo e no consumo de recursos entre os centros. Em comparação com o reembolso nacional de melhores práticas, o custo médio por cirurgia de prótese de quadril de paciente internado de todos os seis hospitais foi de aproximadamente 7x o reembolso nacional. Conclusão: A aplicação do TDABC nos permitiu identificar diferenças na via de atendimento cirúrgico entre hospitais, o que poderia ser explorado em estudos futuros que visem projetar uma via cirúrgica de referência. As diferenças na forma como o tratamento está sendo entregue aos pacientes também contribuíram para justificar a alta variabilidade dos custos entre os centros.

Lessons from Brazil's unsuccessful fiscal decentralization policy to fight COVID-19.

This manuscript investigates the unsuccessful case of the fiscal decentralization policy implemented by the Brazilian central government to help municipalities fight COVID-19. Based on quantitative analyses of data available on governmental websites, we identified that the transfer policy had ignored municipalities' risk patterns and income changes. It benefited municipalities regardless of their vulnerability and population infection risks, and many municipalities reduced healthcare expenditures funded by their revenues during the pandemic. Hence, some municipalities made a "pandemic surplus" in 2020 - a municipal electoral year. Indeed, COVID-19 killed 663,694 people in Brazil until 4 May 2022. Lessons from an unsuccessful case of response to COVID-19 help develop resilience for other crises by emerging market economies and developing countries. The findings have implications for policymakers and literature since they represent inadequate vertical coordination that followed a path dependence on traditional decentralization policies and took place in a year of municipal elections without clear spending and accountability rules.

Efeitos políticos na descontinuidade do orçamento participativo em municípios / Efectos políticos sobre la discontinuidad del presupuesto participativo en los municipios / Political effects on the discontinuation of participatory budgeting in municipalities

Resumo O artigo analisa fatores associados à descontinuidade do orçamento participativo (OP) em municípios brasileiros acima de 50 mil habitantes no período entre 2000 e 2016. A pesquisa foi analisada pela estimação de modelos econométricos com foco em variáveis explicativas políticas e pela propensão do ambiente local à participação. Os resultados indicam que a descontinuidade está associada positivamente com a troca do partido do prefeito municipal entre as eleições, a despeito da ideologia do partido; positivamente com a troca da ideologia do partido do prefeito, com a entrada de partidos de esquerda; negativamente com a entrada de partidos com outras ideologias; e positivamente com a troca entre partidos de esquerda. A descontinuidade não apresentou efeito com a troca entre partidos com outras ideologias. Os resultados indicam fatores locais e contextuais explicativos para a descontinuidade do OP, permitindo direcionar desenho de políticas públicas participativas e levantando reflexividade sobre mecanismos que devem ser adotados localmente para que não ocorra a descontinuidade, além de contribuir para a literatura ao demonstrar a associação entre a descontinuidade com a troca de partidos da esquerda, o que pode suscitar pesquisas futuras, dado que o resultado não era esperado.

Resumen El artículo analizó factores asociados a la discontinuidad del presupuesto participativo (PP) en municipios brasileños con más de 50.000 habitantes, en el período 2000-2016. La discontinuidad se analizó mediante la estimación de modelos econométricos con enfoque en variables políticas explicativas y la propensión del entorno local a participar. Los resultados indican que la discontinuidad se asocia: (i) positivamente con el cambio de partido del alcalde entre elecciones, independientemente de la ideología del partido; (ii) positivamente con el cambio de ideología del partido de la alcaldía, con la entrada de partidos de izquierda; (iii) negativamente con la entrada de partidos con otras ideologías; (iv) positivamente con el intercambio entre partidos de izquierda. La discontinuidad no tuvo efecto con el intercambio entre partidos con otras ideologías. Los resultados contribuyen al señalar factores locales y contextuales que explican la discontinuidad del PP, permitiendo orientar el diseño de políticas públicas participativas y planteando la reflexión sobre los mecanismos que deben ser adoptados localmente para que no ocurra la discontinuidad. También contribuye a la literatura al demostrar la asociación entre la discontinuidad y el intercambio de partidos de izquierda, lo que puede impulsar futuras investigaciones, dado que el resultado no era el esperado.

Abstract The article analyzes the factors associated with the discontinuation of participatory budgeting (PB) in Brazilian municipalities with more than 50,000 inhabitants between 2000 and 2016. We used econometric models to estimate PB's discontinuation based on political explanatory variables and people's local participation. The results indicate that discontinuation is associated: (i) positively with the election of a different local government, regardless of the winning party's ideology; (ii) positively with the election of a local government with a left-wing ideology'; (iii) negatively with the election of a non-left-wing local government; (iv) positively with the election of a left-wing local government replacing another left-wing government. Changes in local government where other non-left-wing parties replace non-left-wing parties had no significant effect on discontinuation. The results contribute to our knowledge by showing the local and contextual factors that explain the discontinuation of participatory budgeting; this should help guide the design of participatory public policies and allow practitioners to reflect on the mechanisms that could be adopted locally to avoid discontinuation. The article also contributes to the literature by demonstrating the association between discontinuation and changes in left-wing local governments, which may prompt future research, given the unexpected result.

SARS-CoV-2 productively infects primary human immune system cells in vitro and in COVID-19 patients.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is associated with a hyperinflammatory state and lymphocytopenia, a hallmark that appears as both signature and prognosis of disease severity outcome. Although cytokine storm and a sustained inflammatory state are commonly associated with immune cell depletion, it is still unclear whether direct SARS-CoV-2 infection of immune cells could also play a role in this scenario by harboring viral replication. We found that monocytes, as well as both B and T lymphocytes, were susceptible to SARS-CoV-2 infection in vitro, accumulating double-stranded RNA consistent with viral RNA replication and ultimately leading to expressive T cell apoptosis. In addition, flow cytometry and immunofluorescence analysis revealed that SARS-CoV-2 was frequently detected in monocytes and B lymphocytes from coronavirus disease 2019 (COVID-19) patients. The rates of SARS-CoV-2-infected monocytes in peripheral blood mononuclear cells from COVID-19 patients increased over time from symptom onset, with SARS-CoV-2-positive monocytes, B cells, and CD4+ T lymphocytes also detected in postmortem lung tissue. These results indicated that SARS-CoV-2 infection of blood-circulating leukocytes in COVID-19 patients might have important implications for disease pathogenesis and progression, immune dysfunction, and virus spread within the host.

Time-driven activity-based costing as a strategy to increase efficiency: An analyses of interventional coronary procedures.

Monitoring the costs is one of the key components underlying value-based health care. This study aimed to evaluate the cost-saving opportunities of interventional coronary procedures (ICPs). Data from 90 patients submitted to elective ICP were evaluated in five Brazilian hospitals. Time-driven activity-based costing, that guides the cost estimates using the time consumed and the capacity cost rates per resource as the data input, was used to assess costs and the time spent over the care pathway. Descriptive cost analyses were followed by a labour cost-saving estimate potentially achieved by the redesign of the ICP pathway. The mean cost per patient varied from $807 to $2639. The length of the procedure phase per patient was similar among the hospitals, while the post-procedure phase presented the highest variation in length. The highest direct cost saving opportunities are concentrated in the procedure phase. By comparing the benchmark service with the most expensive one, it was estimated that redesigning physician practices could decrease 51% of the procedure cost. This application is pioneered in Brazil and demonstrates how detailed cost information can contribute to driving health care management to value by identifying cost-saving opportunities.

Neural Infection by Oropouche Virus in Adult Human Brain Slices Induces an Inflammatory and Toxic Response.

Oropouche virus (OROV) is an emerging arbovirus in South and Central Americas with high spreading potential. OROV infection has been associated with neurological complications and OROV genomic RNA has been detected in cerebrospinal fluid from patients, suggesting its neuroinvasive potential. Motivated by these findings, neurotropism and neuropathogenesis of OROV have been investigated in vivo in murine models, which do not fully recapitulate the complexity of the human brain. Here we have used slice cultures from adult human brains to investigate whether OROV is capable of infecting mature human neural cells in a context of preserved neural connections and brain cytoarchitecture. Our results demonstrate that human neural cells can be infected ex vivo by OROV and support the production of infectious viral particles. Moreover, OROV infection led to the release of the pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-α) and diminished cell viability 48 h post-infection, indicating that OROV triggers an inflammatory response and tissue damage. Although OROV-positive neurons were observed, microglia were the most abundant central nervous system (CNS) cell type infected by OROV, suggesting that they play an important role in the response to CNS infection by OROV in the adult human brain. Importantly, we found no OROV-infected astrocytes. To the best of our knowledge, this is the first direct demonstration of OROV infection in human brain cells. Combined with previous data from murine models and case reports of OROV genome detection in cerebrospinal fluid from patients, our data shed light on OROV neuropathogenesis and help raising awareness about acute and possibly chronic consequences of OROV infection in the human brain.

Spatiotemporal variability of solid waste on sandy beaches with different access restrictions.

Intense human use and high construction density in coastal areas are stressors to sandy beaches. Pollution by marine debris is a major problem on beaches worldwide. This study pioneered an assessment of marine debris characterization over time on beaches with different levels of access. In two periods and seasons, marine debris was sampled on nine sandy beaches of Rio de Janeiro, grouped by levels of access. The general marine debris density has decreased over time, accompanied by an improvement in public cleaning mechanisms. The most important predictor for the majority of marine debris items is related to accessibility; beaches with restricted access showed a reduction in the abundance of most items. High marine debris densities, even on beaches with restricted access, showed that all evaluated beaches can be sinks for marine debris circulating in marine waters. Beach cleaning strategy improvements will be inefficient unless integrated marine debris management is implemented.

Detection of respiratory viruses in primary cholesteatoma tissues.

Cholesteatomas are frequent middle ear benign tumors of unknown etiology. Infectious agents have been considered as possible contributing factors in the pathogenesis of cholesteatomas. Aiming to investigate the presence of respiratory viruses in primary cholesteatoma tissues, 26 formalin-fixed paraffin-embedded primary cholesteatoma tissues obtained from patients seen at the of the Clinical Hospital of the University of São Paulo School of Medicine, in Ribeirão Preto, Brazil were tested by real-time polymerase chain reaction (PCR). Considering the PCR results, 35% of the tissues were positive for human rhinovirus (HRV), 15.3% for human enterovirus (EV), 3.8% for human metapneumovirus (HMPV), and 3.8% for human bocavirus (HBoV). Serial immunohistochemistry for virus antigens and cell surface markers evidenced that the viruses were associated with fibroblasts, dendritic cells, macrophages, B lymphocytes, CD4+ , and CD8+ T lymphocytes. These findings indicate for the first time the presence of active respiratory virus infection in primary cholesteatoma tissues, suggesting that persisting virus infection in the middle could play a role in the pathogenesis and evolution of cholesteatomas.

The Economic Impact of COVID-19 Treatment at a Hospital-level: Investment and Financial Registers of Brazilian Hospitals.

Background: The economic impact associated with the treatment strategies of coronavirus disease-2019 (COVID-19) patients by hospitals and health-care systems in Brazil is unknown and difficult to estimate. This research describes the investments made to absorb the demand for treatment and the changes in occupation rates and billing in Brazilian hospitals. Methods: This research covers the initial findings of "COVID-19 hospital costs and the proposition of a bundled reimbursement strategy for the health-care system," which includes 10 hospitals. The chief financial officer, the chief medical officer, and hospital executives of each participating hospital provided information regarding investments attributed to COVID-19 patient treatment. The analysis included variations in occupation rates and billing from 2019 to 2020 observed in each institution, and the investments for medical equipment, individual protection materials and building construction per patient treated. Results: The majority of hospitals registered a decrease in hospitalization rates and revenue from 2019 to 2020. For intensive care units (ICUs), the mean occupancy rate ranged from 88% to 83%, and for wards, it ranged from 85% to 73%. Monthly average revenue decreased by 10%. The mean hospital investment per COVID-19 inpatient was I$6800 (standard deviation 7664), with the purchase of ventilators as the most common investment. For this item, the mean, highest and lowest acquisition cost per ventilator were, respectively, I$31 468, I$48 881 and I$17 777. Conclusion: There was significant variability in acquisition costs and investments by institution for responding to the COVID-19 pandemic. These findings highlight the importance of continuing microeconomic studies for a comprehensive assessment of hospital costs. Only with more detailed analyses, will it be possible to define and drive sustainable strategies to manage and reimburse COVID-19 treatment in health-care systems.

Human Respiratory Syncytial Virus Infection in a Human T Cell Line Is Hampered at Multiple Steps.

Human respiratory syncytial virus (HRSV) is the most frequent cause of severe respiratory disease in children. The main targets of HRSV infection are epithelial cells of the respiratory tract, and the great majority of the studies regarding HRSV infection are done in respiratory cells. Recently, the interest on respiratory virus infection of lymphoid cells has been growing, but details of the interaction of HRSV with lymphoid cells remain unknown. Therefore, this study was done to assess the relationship of HRSV with A3.01 cells, a human CD4+ T cell line. Using flow cytometry and fluorescent focus assay, we found that A3.01 cells are susceptible but virtually not permissive to HRSV infection. Dequenching experiments revealed that the fusion process of HRSV in A3.01 cells was nearly abolished in comparison to HEp-2 cells, an epithelial cell lineage. Quantification of viral RNA by RT-qPCR showed that the replication of HRSV in A3.01 cells was considerably reduced. Western blot and quantitative flow cytometry analyses demonstrated that the production of HRSV proteins in A3.01 was significantly lower than in HEp-2 cells. Additionally, using fluorescence in situ hybridization, we found that the inclusion body-associated granules (IBAGs) were almost absent in HRSV inclusion bodies in A3.01 cells. We also assessed the intracellular trafficking of HRSV proteins and found that HRSV proteins colocalized partially with the secretory pathway in A3.01 cells, but these HRSV proteins and viral filaments were present only scarcely at the plasma membrane. HRSV infection of A3.01 CD4+ T cells is virtually unproductive as compared to HEp-2 cells, as a result of defects at several steps of the viral cycle: Fusion, genome replication, formation of inclusion bodies, recruitment of cellular proteins, virus assembly, and budding.

Host Retromer Protein Sorting Nexin 2 Interacts with Human Respiratory Syncytial Virus Structural Proteins and is Required for Efficient Viral Production.

Human respiratory syncytial virus (HRSV) envelope glycoproteins traffic to assembly sites through the secretory pathway, while nonglycosylated proteins M and N are present in HRSV inclusion bodies but must reach the plasma membrane, where HRSV assembly happens. Little is known about how nonglycosylated HRSV proteins reach assembly sites. Here, we show that HRSV M and N proteins partially colocalize with the Golgi marker giantin, and the glycosylated F and nonglycosylated N proteins are closely located in the trans-Golgi, suggesting their interaction in that compartment. Brefeldin A compromised the trafficking of HRSV F and N proteins and inclusion body sizes, indicating that the Golgi is important for both glycosylated and nonglycosylated HRSV protein traffic. HRSV N and M proteins colocalized and interacted with sorting nexin 2 (SNX2), a retromer component that shapes endosomes in tubular structures. Glycosylated F and nonglycosylated N HRSV proteins are detected in SNX2-laden aggregates with intracellular filaments projecting from their outer surfaces, and VPS26, another retromer component, was also found in inclusion bodies and filament-shaped structures. Similar to SNX2, TGN46 also colocalized with HRSV M and N proteins in filamentous structures at the plasma membrane. Cell fractionation showed enrichment of SNX2 in fractions containing HRSV M and N proteins. Silencing of SNX1 and 2 was associated with reduction in viral proteins, HRSV inclusion body size, syncytium formation, and progeny production. The results indicate that HRSV structural proteins M and N are in the secretory pathway, and SNX2 plays an important role in the traffic of HRSV structural proteins toward assembly sites.IMPORTANCE The present study contributes new knowledge to understand HRSV assembly by providing evidence that nonglycosylated structural proteins M and N interact with elements of the secretory pathway, shedding light on their intracellular traffic. To the best of our knowledge, the present contribution is important given the scarcity of studies about the traffic of HRSV nonglycosylated proteins, especially by pointing to the involvement of SNX2, a retromer component, in the HRSV assembly process.