Ultrastructural changes induced by açaí (Euterpe oleracea Mart) in MCF-7 breast cancer cell line.

Açaí (Euterpe oleracea Mart) is an Amazon plant with many biological properties. Previous report of this group evidenced autophagy induction after treatment with açaí seed extract in MCF-7 breast cancer cell lines by acridine orange assay. The aim of this study was to evaluate the ultrastructural changes induced by açaí seed extract in MCF-7 breast cancer cell lines. First, MCF- 7 breast cancer cell line viability was evaluated by MTT assay. Acridine orange assay showed increase in the acidic compartments, suggesting autophagolysosome formation. These cells were treated with 25 µg/ml for 24 h and evaluated by transmission electron microscopy (MET). This analysis showed that açaí seed extract induced autophagy, confirmed by autophagolysosome formation. Furthermore, açaí seed extract increased the number of mitochondria, suggesting the enrollment of reactive oxygen species in autophagy.

Impact of Delirium and Its Motor Subtypes on Stroke Outcomes.

BACKGROUND AND PURPOSE: Delirium is an acute and fluctuating impairment of attention, cognition, and behavior. Although common in stroke, studies that associate the clinical subtypes of delirium with functional outcome and death are lacking. We aimed to evaluate the influence of delirium occurrence and its different motor subtypes over stroke patients' prognosis. METHODS: Prospective cohort of stroke patients with symptom onset within 72 hours before research admission. Delirium was diagnosed by Confusion Assessment Method for the Intensive Care Unit, and its motor subtypes were defined according to the Richmond Agitation-Sedation Scale. The main outcome was functional dependence or death (modified Rankin Scale>2) at 90 days comparing: delirium versus no delirium patients; and between motor subtypes. Secondary outcomes included modified Rankin Scale score >2 at 30 days and 90-day-mortality. RESULTS: Two hundred twenty-seven patients were enrolled. Delirium occurred in 71 patients (31.3%), with the hypoactive subtype as the most frequent, in 41 subjects (57.8%). Delirium was associated with increased risk of death and functional dependence at 30 and 90 days and higher 90-day mortality. Multivariate analysis showed delirium (odds ratio, 3.28 [95% CI, 1.17-9.22]) as independent predictor of modified Rankin Scale >2 at 90 days. CONCLUSIONS: Delirium is frequent in stroke patients in the acute phase. Its occurrence-specifically in mixed and hypoactive subtypes-seems to predict worse outcomes in this population. To our knowledge, this is the first study to prospectively investigate differences between delirium motor subtypes over functional outcome three months poststroke. Larger studies are needed to elucidate the relationship between motor subtypes of delirium and functional outcomes in the context of acute stroke.

A longitudinal study of convalescent plasma (CCP) donors and correlation of ABO group, initial neutralizing antibodies (nAb), and body mass index (BMI) with nAb and anti-nucleocapsid (NP) SARS-CoV-2 antibody kinetics: Proposals for better quality of CCP collections.

INTRODUCTION: Little is known about the neutralizing (nAb) and binding antibody kinetics in COVID-19 convalescent plasma donors, especially during the first 100 days after disease onset. MATERIALS AND METHODS: A cohort of previously RT-PCR positive (detected by nasopharyngeal swab during the acute phase), male convalescent patients, all with mild symptoms, were enrolled in serial blood sample collection for a longitudinal nAb titers and anti-nucleocapsid (NP) antibodies (IgM, IgG and IgA) evaluation. NAbs were detected by a cytopathic effect-based virus neutralization test (CPE-based VNT), carried out with SARS-CoV-2 (GenBank: MT350282). RESULTS: A total of 78 male volunteers provided 316 samples, spanning a total of 4820 days of study. Although only 25% of donors kept nAb titers ≥160 within 100 days after the onset of disease, there was >75% probability of sustaining nAb titers ≥160 in volunteers whose initial nAb titer was ≥1280, weight ≥ 90 kg or obese, according to their body mass index (BMI), as evidenced by Kaplan-Meier analysis and Cox hazard regression (all p < .02). There was no correlation between the ABO group, ABO antibody titers and persistent high nAb titers. High IgG anti-NP (S/CO ≥5.0) is a good surrogate for detecting nAb ≥ 160, defined by the ROC curve (sensitivity = 90.5%; CI95%: 84.5%-94.7%). CONCLUSION: Selection of CCP donors for multiple collections based on initial high nAb titers (≥1280) or BMI ≥ 30 kg/m2 provides a simple strategy to achieve higher quality in CCP programs. High IgG anti-NP levels can also be used as surrogate markers for high nAb screening.

Surrogate test performance for SARS-CoV-2 neutralizing antibodies (nAbs) for convalescent plasma (CCP): How useful could they be?

BACKGROUND: COVID-19 high-titer CCP selection is a concern, because neutralizing antibody (nAb) testing requires sophisticated labs and methods. Surrogate tests are an alternative for measuring nAb levels in plasma bags, including those that are pathogen-reduced. STUDY DESIGN/METHODS: We studied a panel consisting of 191 samples from convalescent donors tested by nAb (CPE-VNT), obtained from 180 CCP donations (collection: March 20-January 21) and 11 negative controls, with a total of 80 and 111 serum and plasma samples (71 amotosalen/UV treated), with nAb titers ranging from negative to 10,240. Samples were blindly tested for several surrogates: one anti-RBD, two anti-spike, and four anti-nucleocapsid tests, either isolated or combined to improve their positive predictive values as predictors of the presence of high-titer nAbs, defined as those with titers ≥160. RESULTS: Except for combined and anti-IgA/M tests, all isolated surrogate tests showed excellent performance for nAb detection: sensitivity (98.3%-100%), specificity (85.7%-100%), PPV (98.9%-100%), NPV (81.3%-100%), and AUC (0.93-0.96), with a variable decrease in sensitivity and considerably lower specificity when using FDA authorization and concomitant nAb titers ≥160. All surrogates had AUCs that were statistically different from CPE-VNT if nAb≥160, including when using combined, orthogonal approaches. CONCLUSIONS: Surrogate tests (isolated or in combination) have an indirect good performance in detecting the presence of nAb, with lower sensitivity and specificity when high nAb titer samples are used, possibly accepting a considerable number of donors whose nAb titers are actually low, which should be evaluated by each laboratory responsible for CCP collection.

Anti-A and SARS-CoV-2: an intriguing association.

BACKGROUND: Blood groups and anti-A isohemagglutinin may be involved in susceptibility to SARS-CoV-2 infection. MATERIALS AND METHODS: We retrospectively studied 268 COVID-19 convalescent plasma donors and 162 COVID-19 inpatients (total 430 subjects, confirmed by RT-PCR) and 2,212 healthy volunteer first-time blood donors as a control group. These were further divided into two groups: those with anti-A (blood types O and B) and those without it (types A and AB). Titres of nucleoproteins, and neutralizing SARS-CoV-2 antibody were measured in the convalescent plasma donors and inpatients. Multivariate logistic regression and non-parametric tests were applied. RESULTS: Persons having types O or B showed less infection prevalence than those of types A or AB (OR = 0·62, 95% CI 0·50-0·78; P < 0·001), but there was no difference when COVID-19 inpatients were analysed. Immunoglobulins M, G and A were lower in COVID-19 subjects of types O or B group than those of A or AB (0·16 vs. 0·19; P = 0·03, 2·11 vs. 2·55; P = 0·02, 0·23 vs. 0·32; P = 0·03, respectively). CONCLUSION: In this retrospective cohort, COVID-19 individuals were less likely to belong to blood types O and B, and also had lower SARS-CoV-2 antibody titres than A and AB individuals. COVID-19 severity did not associate with the blood groups.

Screening for SARS-CoV-2 antibodies in convalescent plasma in Brazil: Preliminary lessons from a voluntary convalescent donor program.

BACKGROUND: Coronavirus disease 2019 (COVID-19) convalescent plasma (CCP) collection began in two Brazilian hospitals for treatment of severe/critical patients. METHODS AND MATERIALS: Mild/moderate COVID-19 convalescents were selected as CCP donors after reverse transcription polymerase chain reaction (RT-PCR) confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and absence of symptoms for ≥14 days plus (a) age (18-60 years), body weight greater than 55 kg; (b) immunohematological studies; (c) no infectious markers of hepatitis B virus, hepatitis C virus, human immunodeficiency virus, human T-lymphotropic virus-1/2, Chagas and syphilis infection; (d) no HLA antibodies (multiparous); (e) second RT-PCR (nasopharyngeal swab and/or blood) negativity; (f) virus neutralization test (cytopathic effect-based virus neutralization test neutralizing antibody) and anti-nucleocapsid protein SARS-CoV-2 IgM, IgG, and IgA enzyme-linked immunosorbent assays. RESULTS: Among 271 donors (41 females, 230 males), 250 presented with neutralizing antibodies. Final RT-PCR was negative on swab (77.0%) or blood (88.4%; P = .46). Final definition of RT-PCR was only defined at more than 28 days after full recovery in 59 of 174 (33.9%) RT-PCR -ve, and 25/69 RT-PCR +ve (36.2%; 13 between 35 and 48 days). Neutralizing antibody titers of 160 or greater were found in 63.6%. Correlation between IgG signal/cutoff of 5.0 or greater and neutralizing antibody of 160 or greater was 82.4%. Combination of final RT-PCR -ve with neutralizing antibody ≥160 was 41.3% (112/271). Serial plasma collection showed decline in neutralizing antibody titers and IgA levels (P < .05), probably denoting a "golden period" for CCP collection (≤28 days after joining the program); IgA might have an important role as neutralizing antibody. Donor's weight, days between disease onset and serial plasma collection, and IgG and IgM levels are important predictors for neutralizing antibody titer. CONCLUSIONS: RT-PCR +ve cases are still detected in 36.2% within 28 to 48 days after recovery. High anti-nucleocapsid protein IgG levels may be used as a surrogate marker to neutralizing antibody.

High prevalence of SARS-CoV-2 infection among symptomatic healthcare workers in a large university tertiary hospital in São Paulo, Brazil.

BACKGROUND: Brazil became the epicenter of the COVID-19 pandemic in Latin America since May 2020, reporting the highest number of cases and deaths in the region. Healthcare workers (HCWs) are at increased risk of SARS-CoV-2 infection, experiencing a significant burden from COVID-19. Identifying and understanding the clinical characteristics and risk factors associated with infection are of paramount importance to inform screening strategies and infection control practices in this scenario. The aims of this study were to investigate the prevalence and clinical characteristics of HCWs with COVID-19 symptoms. METHODS: Between March 21st and May 22nd, 2020 a cross-sectional study was performed in a tertiary university hospital in São Paulo. Prevalence of SARS-CoV-2 infection among HCWs with COVID-19 symptoms was determined by RT-PCR testing on nasopharyngeal and oropharyngeal samples. Participants were asked to complete an electronic structured questionnaire including clinical and demographic data. RESULTS: Overall, 125 (42.37%) of 295 symptomatic HCWs tested positive for SARS-CoV-2. Over the 10-week study period, positivity rates varied from 22.2% (95% CI 15.9-60.3%) in the second week to 55.9% (95% CI 43.2-68.6%) in the sixth week, reaching a plateau (38-46%) thereafter. Median (SD) age was 34.2 (9.9) years and 205 (69.5%) were female. We did not find significant differences in the prevalence of the most commonly reported underlying medical condition among healthcare workers that tested positive or negative for SARS-CoV-2 infection. After multivariable analysis, using logistic regression, anosmia (adjusted OR 4.4 95% CI 2.21-8.74) and ocular pain (adjusted OR 1.95 95% CI 1.14-3.33) were the only symptoms independently associated with positivity for SARS-CoV-2 infection. Follow-up information on clinical outcomes showed that 9 (7.2%) HCWs were hospitalized (seven were male) and 2 (1.6%) died. CONCLUSIONS: The findings of this study confirmed the high burden of SARS-CoV-2 infection among healthcare workers in the hardest hit city by the pandemic in Latin America. Anosmia and ocular pain were symptoms independently associated with COVID-19 diagnosis. In low and middle-income countries, where limited availability of tests is frequent, these findings may contribute to optimize a targeted symptom-oriented screening strategy.

SARS-CoV-2 isolation from the first reported patients in Brazil and establishment of a coordinated task network.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was confirmed in Brazil in February 2020, the first cases were followed by an increase in the number of cases throughout the country, resulting in an important public health crisis that requires fast and coordinated responses. OBJECTIVES: The objective of this work is to describe the isolation and propagation properties of SARS-CoV-2 isolates from the first confirmed cases of coronavirus disease 2019 (COVID-19) in Brazil. METHODS: After diagnosis in patients that returned from Italy to the São Paulo city in late February by RT-PCR, SARS-CoV-2 isolates were obtained in cell cultures and characterised by full genome sequencing, electron microscopy and in vitro replication properties. FINDINGS: The virus isolate was recovered from nasopharyngeal specimen, propagated in Vero cells (E6, CCL-81 and hSLAM), with clear cytopathic effects, and characterised by full genome sequencing, electron microscopy and in vitro replication properties. Virus stocks - viable (titre 2.11 × 106 TCID50/mL, titre 1.5 × 106 PFUs/mL) and inactivated from isolate SARS.CoV2/SP02.2020.HIAE.Br were prepared and set available to the public health authorities and the scientific community in Brazil and abroad. MAIN CONCLUSION: We believe that the protocols for virus growth and studies here described and the distribution initiative may constitute a viable model for other developing countries, not only to help a rapid effective pandemic response, but also to facilitate and support basic scientific research.

Combinations of cactus pear with different roughage sources on the production, chemical composition, and milk fatty acid profile of F1 Holstein/Zebu cows.

The qualities of food, mainly of animal origin, have always been of concern to consumers. It is known that the composition of animals' diets can influence the composition of the foods produced, such as milk. Thus, the objective of this study was to evaluate diets with cactus pear in combination with different forages for F1 Holstein/Zebu cows on the chemical composition and fatty acid profile of milk. Two simultaneous 4 × 4 Latin squares were used in the experimental design. Four experimental diets were used: Diet 1, sorghum silage as the only roughage; Diet 2, sorghum silage combined with cactus pear in a proportion of 50% of the roughage (dry matter basis); Diet 3, elephant grass (Pennisetum purpureum cv. Roxo) as the only roughage; and Diet 4, elephant grass combined with cactus pear in a proportion of 50% of the roughage. The roughage:concentrate ratio was 75:25. The milk urea nitrogen was higher (16.08 mg/dL) in the milk of cows fed the diet with only elephant grass than that of milk from the other diet groups. The other analyzed variables of the chemical composition of milk were not influenced by the diets. However, there was a change in the sum of the saturated and monounsaturated fatty acids. The content of C16:0 fatty acid was higher and that of C18:0 was lower in the milk of cows fed the cactus pear diets than in that of cows fed the other diets. Higher levels of oleic acid were observed in the milk of cows fed with diets containing elephant grass than those in the milk of the cows in the sorghum forage groups. Conjugated linoleic acid (CLA) was higher in the milk of cows fed cactus pear than in that of the other cows. The combination of cactus pear with elephant grass or sorghum silage in the diet did not alter the chemical composition of milk. However, it influenced the fatty acid profile.

Nutritional and productive parameters of Holstein/Zebu cows fed diets containing cactus pear

Objective: This study ascertained effects of cactus pear in association with different roughage in the diet of F1 Holstein/Zebu cows on intake, nutrient digestibility, nitrogen balance, ingestive behavior and performance. Methods: Eight cows with 72 ± 11 days of lactation were used. The experimental design was simultaneous in two 4 x 4 Latin squares. Four experimental diets were used: Diet 1 - sorghum silage as exclusive roughage; Diet 2 - sorghum silage associated with cactus pear in a proportion of 50% of the roughage (dry basis); Diet 3 - elephant grass (Pennisetum purpureum cv. Roxo) as exclusive roughage; Diet 4 - elephant grass associated with cactus pear in a proportion of 50% of the roughage. The roughage:concentrate ratio was 75:25. Dry matter intake (P = 0.01) was higher with sorghum silage. Results: There were differences in dry matter intake (P = 0.01), crude protein (P <0.01), ether extract (P = 0.01), non-fibrous carbohydrates (P <0.01) and total digestible nutrients (P = 0.01) among the diets. Cactus pear in the diet reduced water intake by 44.52% (P < 0.01). The nitrogen balance was 59.71% and 27.49% lower in animals treated with exclusive sorghum silage and sorghum silage associated with cactus pear in relation to diets with elephant grass and elephant grass associated with cactus pear, respectively (P < 0.01). The diets did not influence the milk production (P = 0.70), 3.5% fat corrected milk production (P = 0.72) or feed efficiency (P = 0.61). Conclusion: The association of cactus pear with sorghum or elephant grass silage does not alter milk production, reduces the intake of dry matter and water and improves the digestibility of nutrients.

Histopathology and changes in the expression of metallothioneins, heat shock proteins and inducible nitric oxide synthase in Prochilodus costatus from a neotropical river contaminated by heavy metals.

The most recent dam rupture in Brazil released tons of mining tailings into the upper course of the Paraopeba River, affecting this river in an unprecedented way. The present study aimed to evaluate the influence of heavy metals on Prochilodus costatus, an important commercial species in Brazil, four years after the dam colapse. To this end, biomarkers of heavy metals, oxidative stress, and environmental stress were analyzed, and histological analyses of target organs were performed. The results demonstrated critical contamination of fish from the Paraopeba River. Increased expression of Metallothioneins - MTs, Heat Shock Protein - HSP70, and inducible nitric oxide synthase - iNOS, as well as greater rates of histological changes in the liver, spleen, and gonads, were observed in P. costatus. These findings demonstrate that, despite past contamination, the metals present in mining tailings have significantly increased the contamination of the Paraopeba River basin.

Lifestyle, meal times, and sleep patterns changes in higher education professors during COVID-19: Association with non-communicable chronic diseases.

BACKGROUND: In response to the COVID-19 pandemic, educational institutions had to swiftly adapt and transition to remote teaching in order to maintain academic activities. However, these changes presented a number of challenges for professors, which could have negative effects on their health. OBJECTIVE: To analyze the association between changes in dietary and sleep habits, physical activity level, and sedentary behavior with the development of non-communicable diseases (NCDs) among Brazilian higher education professors during the pandemic period. METHODS: This is a cross-sectional and retrospective study conducted using an online form. Generalized linear models, adjusted for age, sex, and body mass index, were used to verify the difference between pre-pandemic and pandemic periods. Logistic regression models were used to predict the odds ratio (OR) for the development of NCDs according to physical activity time, sedentary behavior time, dietary and sleep patterns. RESULTS: A total of 936 professors residing across Brazil participated in the survey. The duration of sedentary behavior increased, sleep duration slightly decreased, and meal times shifted to earlier during the pandemic. A total of 22.9%of the participants reported the diagnosis of some NCDs during this period. Physical activity practice was associated with a lower risk of diseases during the pandemic, regardless of the intensity performed. On the other hand, late eating habits and excessive food consumption during the pandemic were associated with a higher risk. CONCLUSION: The results provide data that can help in the development of public policies that promote health actions to minimize the consequences associated with the pandemic period.

Preterm births prevalence during the COVID-19 pandemic in Brazil: results from the national database.

The SARS-CoV-2 (COVID-19) pandemic impacted the health systems between and within countries, and in the course of the pandemic sexual and reproductive health services were the most disrupted. Findings from high-income settings have reported significant changes in preterm birth prevalence during the pandemic period. To understand the possible effects of the COVID-19 pandemic on preterm birth numbers at the Brazilian national level. We compare the number of preterm deliveries during the COVID-19 pandemic period (2020 and 2021) with previous years. We conducted a population-based cross-sectional study taking the period from January 2017 to December 2021 to account. We use individual-level live births data from the Brazilian Live Birth Information System (SINASC), and we estimate the odds ratio (OR) of preterm deliveries using propensity score weighting analysis in Brazil and its regions. During the study period (from 2017 to 2021), about 2.7 million live births were recorded per year, and the missing value for gestational age at delivery was less than 1.5%. The preterm birth prevalence slightly increased during the COVID-19 pandemic compared to the pre-pandemic period (11.32% in 2021 vs 11.09% in 2019, p-value < 0.0001). After adjusting for sociodemographic variables, the OR of preterm births in Brazil has significantly increased, 4% in 2020 (OR: 1.04 [1.03-1.05] 95% CI, p-value < 0.001), and 2% in 2021(OR: 1.02 [1.01-1.03] 95% CI, p-value < 0.001), compared to 2019. At the regional level, the preterm birth pattern in the South, Southeast and Northeast regions show a similar pattern. The highest odds ratio was observed in the South region (2020 vs 2019, OR: 1.07 [1.05-1.10] 95% CI; 2021 vs 2019, OR: 1.03 [1.01-1.06] 95% CI). However, we also observed a significant reduction in the ORs of preterm births in the northern region during the COVID-19 pandemic (2020 vs 2019, OR: 0.96 [0.94-0.98] 95% CI) and (2021 vs 2019, OR: 0.97 [0.95-0.99] 95% CI). Our analysis shows that the pandemic has increased regional variation in the number of preterm births in Brazil in 2020 and 2021 compared to the pre-pandemic years.

Ergonomic interventions for work in a sitting position: an integrative review.

The sitting position is one of the most common positions in the workplace and one that can contribute to overloading the musculoskeletal system. Ergonomics can play a significant role in ensuring an appropriate relationship between people and their work and in achieving better conditions for workers' health. The objective of this study was to consult the available evidence on the results of different ergonomic interventions for the musculoskeletal systems of workers who perform their jobs in a sitting position. This was an integrative review, searching the LILACS, MEDLINE, PubMed, SciELO, and CINAHL electronic databases for articles published from 2010 to 2019. The following keywords were used: Trabalhadores OR Workers OR Trabajadores AND Dor OR Pain OR Dolor AND Postura Sentada OR Sitting Position OR Sedestación AND Ergonomia OR Ergonomics OR Ergonomía. A total of 183 articles were identified, 14 of which were selected for the review. For qualitative analysis, the articles were organized by author, year, sample/population, objective, analytical instrument, intervention and type of intervention: combinations of physical exercise programs and postural and ergonomic guidance; different types of guidance and facilitating instruments; or configuration of furniture and use of supporting devices. A quantitative analysis of study quality was conducted using the Physiotherapy Evidence Database, based on the Delphi list. The interventions contributed to improve physical conditions and the tasks being carried out, making them more appropriate for the workers.

A postura sentada é uma das mais adotadas nos ambientes de trabalho e pode contribuir na sobrecarga do sistema musculoesquelético. A ergonomia pode apresentar um papel significativo para manter a relação adequada do homem com o trabalho e para abordar melhores condições à saúde dos trabalhadores. O objetivo deste estudo foi verificar as evidências disponíveis sobre os resultados de diferentes intervenções ergonômicas no sistema musculoesquelético de trabalhadores que exercem atividades na postura sentada. Tratou-se de uma revisão integrativa, utilizando as bases de dados eletrônicas LILACS, MEDLINE, PubMed, SciELO e CINAHL, com busca de artigos publicados no período de 2010 a 2019. Utilizaram-se os seguintes descritores: Trabalhadores OR Workers OR Trabajadores AND Dor OR Pain OR Dolor AND Postura Sentada OR Sitting Position OR Sedestación AND Ergonomia OR Ergonomics OR Ergonomía. Foram encontrados 183 artigos, sendo 14 selecionados. A análise qualitativa organizou os artigos de acordo com autor, ano, amostra/população, objetivo, instrumento de análise, intervenção e tipo de intervenção: associação de programas de exercícios físicos e orientações posturais e ergonômicas; diferentes formas de orientação e instrumentos facilitadores; e configuração do mobiliário e utilização de dispositivos auxiliares. A análise quantitativa da qualidade dos estudos considerou a escala Physiotherapy Evidence Database, baseado na lista Delphi. As intervenções contribuíram para melhorar as condições físicas e as tarefas executadas, tornando-as mais apropriadas para os trabalhadores.

Rehabilitation and release of confiscated songbirds into the wild: A pilot study.

Songbirds are currently the most prevalent animals in illegal trafficking in Brazil and other countries, so they are often confiscated, and this poses legal, ethical, and conservation challenges. Returning them to nature requires complex and expensive management, a topic that is sparingly addressed in the literature. Here, we described the processes and costs associated with an attempt to rehabilitate and release confiscated songbirds into the wild. A total of 1,721 songbirds of several species were quarantined, rehabilitated, and released, primarily on two farms located within their typical geographical distribution. Health assessments were performed on samples from 370 birds. Serology revealed no antibodies against Newcastle disease, and Salmonella spp. cultures were negative. Real-time polymerase chain reactions detected M. gallisepticum in samples from seven birds. Atoxoplasma spp. and Acuaria spp. infections, sepsis, and trauma were the top causes of bird death. About 6% of the released birds were recaptured, within an average period of 249 days after release, and at a mean distance of 2,397 meters from the release sites. The majority of these birds were found with free-living mates within or close to fragments of transitional ecoregions with native or cultivated grasslands, and native groves/forests, and shrublands. However, eucalyptus plantations with rich understory regeneration provided a suitable environment for the released forest species to settle, since they were recaptured during the defense of these sites. Over half of the recaptured birds presented behavioral profiles with both dominant and tame traits. Birds with dominant traits are more likely to settle in habitats and face the live decoys during fieldwork, whereas birds with tame characteristics tend to accept close contact with humans. Ultramarine grosbeak (Cyanoloxia brissonii), the least common species among those released, at the release sites showed an almost 2-fold recapture rate in the shortest mean distances from the release sites. This suggests less territory competition, perhaps a major factor of bird re-establishment here. The total per-bird cost was USD 57. Our findings suggested suitable survival and re-establishment of confiscated songbirds in the wild, when managed as we describe.

SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time Validation.

SARS-CoV-2 diagnostic tests have become an important tool for pandemic control. Among the alternatives for COVID-19 diagnosis, antigen rapid diagnostic tests (Ag-RDT) are very convenient and widely used. However, as SARS-CoV-2 variants may continuously emerge, the replacement of tests and reagents may be required to maintain the sensitivity of Ag-RDTs. Here, we describe the development and validation of an Ag-RDT during an outbreak of the Omicron variant, including the characterization of a new monoclonal antibody (anti-DTC-N 1B3 mAb) that recognizes the Nucleocapsid protein (N). The anti-DTC-N 1B3 mAb recognized the sequence TFPPTEPKKDKKK located at the C-terminus of the N protein of main SARS-CoV-2 variants of concern. Accordingly, the Ag-RDT prototypes using the anti-DTC-N 1B3 mAB detected all the SARS-CoV-2 variants-Wuhan, Alpha, Gamma, Delta, P2 and Omicron. The performance of the best prototype (sensitivity of 95.2% for samples with Ct ≤ 25; specificity of 98.3% and overall accuracy of 85.0%) met the WHO recommendations. Moreover, results from a patients' follow-up study indicated that, if performed within the first three days after onset of symptoms, the Ag-RDT displayed 100% sensitivity. Thus, the new mAb and the Ag-RDT developed herein may constitute alternative tools for COVID-19 point-of-care diagnosis and epidemiological surveillance.

Identification of Variants Responsible for Monogenic Forms of Diabetes in Brazil.

Monogenic forms of diabetes mellitus may affect a significant number of patients of this disease, and it is an important molecular cause to be investigated. However, studies of the genetic causes of monogenic diabetes, especially in populations with mixed ethnic backgrounds, such as the one in Brazil, are scarce. The aim of this study was to screen several genes associated with monogenic diabetes in fifty-seven Brazilian patients with recurrence of the disease in their families and thirty-four relatives. Inclusion criteria were: Age of onset ≤ 40 years old, BMI < 30 kg/m², at least two affected generations and negative anti-GAD and anti-IA2 antibodies. MODY genes HNF4A, GCK, HNF1A, HNF1B, NEUROD1, KLF11, PAX4, INS, KCNJ11, and MT-TL1 were sequenced by Sanger sequencing. We identified a total of 20 patients with variants, 13 GCK-MODY, four HNF1A-MODY, and one variant in each of the following genes, HNF4A, HNF1B and MT-TL1. Segregation analysis was performed in 13 families. Four variants were novel, two in GCK (p.(Met115Val) [c.343A>G] and p.(Asp365GlufsTer95) [c.1094_1095insGCGA]) and two in HNF1A (p.(Tyr163Ter) [c.489C>G] and p.(Val380CysfsTer39) [c.1136_1137insC]). Here we highlight the importance of screening for monogenic diabetes in admixed populations.

Validation of Serological Methods for COVID-19 and Retrospective Screening of Health Employees and Visitors to the São Paulo University Hospital, Brazil.

Reliable serological tests for the detection of SARS-CoV-2 antibodies among infected or vaccinated individuals are important for epidemiological and clinical studies. Low-cost approaches easily adaptable to high throughput screenings, such as Enzyme-Linked Immunosorbent Assays (ELISA) or electrochemiluminescence immunoassay (ECLIA), can be readily validated using different SARS-CoV-2 antigens. A total of 1,119 serum samples collected between March and July of 2020 from health employees and visitors to the University Hospital at the University of São Paulo were screened with the Elecsys® Anti-SARS-CoV-2 immunoassay (Elecsys) (Roche Diagnostics) and three in-house ELISAs that are based on different antigens: the Nucleoprotein (N-ELISA), the Receptor Binding Domain (RBD-ELISA), and a portion of the S1 protein (ΔS1-ELISA). Virus neutralization test (CPE-VNT) was used as the gold standard to validate the serological assays. We observed high sensitivity and specificity values with the Elecsys (96.92% and 98.78%, respectively) and N-ELISA (93.94% and 94.40%, respectively), compared with RBD-ELISA (90.91% sensitivity and 88.80% specificity) and the ΔS1-ELISA (77.27% sensitivity and 76% specificity). The Elecsys® proved to be a reliable SARS-CoV-2 serological test. Similarly, the recombinant SARS-CoV-2 N protein displayed good performance in the ELISA tests. The availability of reliable diagnostic tests is critical for the precise determination of infection rates, particularly in countries with high SARS-CoV-2 infection rates, such as Brazil. Collectively, our results indicate that the development and validation of new serological tests based on recombinant proteins may provide new alternatives for the SARS-CoV-2 diagnostic market.

Development and validation of an enzyme-linked immunoassay kit for diagnosis and surveillance of COVID-19.

There is a massive demand to identify alternative methods to detect new cases of COVID-19 as well as to investigate the epidemiology of the disease. In many countries, importation of commercial kits poses a significant impact on their testing capacity and increases the costs for the public health system. We have developed an ELISA to detect IgG antibodies against SARS-CoV-2 using a recombinant viral nucleocapsid (rN) protein expressed in E. coli. Using a total of 894 clinical samples we showed that the rN-ELISA was able to detect IgG antibodies against SARS-CoV-2 with high sensitivity (97.5%) and specificity (96.3%) when compared to a commercial antibody test. After three external validation studies, we showed that the test accuracy was higher than 90%. The rN-ELISA IgG kit constitutes a convenient and specific method for the large-scale determination of SARS-CoV-2 antibodies in human sera with high reliability.