Acute kidney injury: a post-COVID-19 complication in children and adolescents / Lesão renal aguda: uma complicação pós-COVID-19 em crianças e adolescentes

ABSTRACT Objective: To describe cases of acute kidney injury (AKI) in children diagnosed with COVID-19, associated risk factors, clinical aspects and outcome of cases. Methods: Retrospective study, carried out in a pediatric hospital between March 2020 and September 2021, with patients with COVID-19 who were diagnosed with AKI, studying information present in medical records such as comorbidities, age, gender and use of nephrotoxic medications. Results: We studied 40 cases, and male individuals were significantly more affected (62.5%; p=0.025). AKI was a severe complication of COVID-19 infection, with 100% of the sample requiring admission to the Intensive Care Unit and 22.5% dying. The most prevalent comorbidities analyzed in this study were epilepsy, cerebral palsy and heart disease. Most patients were classified according to Kidney Disease: Improving Global Outcomes (KDIGO) criteria as KDIGO 1 (42.5%), and required orotracheal intubation (67.5%). The frequency of use of nephrotoxic medications and need for dialysis was low, with percentages of 35 and 17.5%, respectively. Among the children who died, 70.4% had some comorbidity and 88.8% received invasive ventilation. Conclusions: AKI in children with COVID-19 infection is associated with severe conditions. Despite the severity, most patients were discharged alive from the hospital.

RESUMO Objetivo: Descrever casos de lesão renal aguda (LRA) em crianças diagnosticadas com COVID-19, associando fatores de risco, aspectos clínicos e evolução dos casos. Métodos: Estudo retrospectivo, realizado em hospital pediátrico entre março de 2020 e setembro de 2021, com pacientes com COVID-19 diagnosticados com LRA, que examinou informações presentes em prontuários como comorbidades, idade, sexo e uso de medicações nefrotóxicas. Resultados: Foram estudados 40 casos, sendo o sexo masculino significativamente mais acometido (62,5%; p=0,025). A LRA foi uma complicação grave da infecção por COVID-19, com 100% da amostra necessitando de internação na Unidade de Terapia Intensiva e 22,5% indo a óbito. As comorbidades mais prevalentes analisadas neste estudo foram epilepsia, paralisia cerebral e cardiopatia. A maioria dos pacientes foi classificada pelos critérios Kidney Disease: Improving Global Outcomes (KDIGO) como KDIGO 1 (42,5%) e necessitou de intubação orotraqueal (67,5%). A frequência de uso de medicamentos nefrotóxicos e necessidade de diálise foi baixa, com percentuais de 35 e 17,5%, respectivamente. Entre as crianças que faleceram, 70,4% apresentavam alguma comorbidade e 88,8% receberam ventilação invasiva. Conclusões: A LRA em crianças com infecção por COVID-19 está associada a quadros graves, apesar de a maior parte dos pacientes ter recebido alta hospitalar.

Choroiditis following severe acute respiratory syndrome coronavirus 2 infection in unvaccinated identical twins

ABSTRACT Unvaccinated identical twins developed bilateral anterior uveitis soon after the onset of coronavirus disease 2019 symptoms. During follow-up, both patients developed choroiditis, and one twine developed posterior scleritis and serous retinal detachment. Prompt treatment with oral prednisone ameliorated the lesions, and no recurrence was observed at the 18-month follow-up. Choroiditis may rarely be associated with severe acute respiratory syndrome coronavirus 2 infection, and it responds well to corticosteroid therapy. Although the exact mechanism is unknown, we hypothesize that the virus may act as an immunological trigger for choroiditis.

Neonatal and maternal outcomes of mRNA versus Non-mRNA COVID-19 vaccines in pregnant patients: a systematic review and meta-analysis.

Objective: To compare the effectiveness and safety of non-mRNA versus mRNA COVID-19 vaccines on pregnant women and their newborns in a systematic review with meta-analysis. Data sources: We searched PubMed, Embase, and Cochrane Central in May 2023. Study selection: The search strategy yielded 4451 results, 16 studies were fully reviewed. We selected case-control studies analysing non-mRNA versus mRNA vaccines. Data collection and analysis: we assessed the risk of bias using the Cochrane Risk of Bias in Non-randomized Studies of Interventions (ROBINS-I) tool. Standardised mean differences were pooled using random-effect models. Data synthesis: We identified 8 prospective and retrospective studies with a total of 32,153 patients. Non-mRNA vaccines were associated with a higher incidence of fever (OR 2.67; 95% CI 2.08-3.43; p<0.001), and a lower incidence of fetal or neonatal death (OR 0.16; 95% CI 0.08-0.33; p<0.001). In subgroup analyses, the Jansen vaccine (Ad26.COV2.S) was found to have a higher rate of premature labor/delivery (OR 4.48; 95% CI 1.45-13.83; p=0.009) and missed/spontaneous abortion (OR 1.90; 95% CI 1.09-3.30; p=0.02), as compared with the Pfizer (BNT162b2) vaccine. Conclusion: non-mRNA vaccines are associated with a lower incidence of fetal or neonatal death among pregnant women who receive a Covid19 vaccine, although at an increased rate of pyrexia compared with mRNA vaccines. Other studies are required for better assessment. PROSPERO: CRD42023421814.

Aging mitochondria in the context of SARS-CoV-2: exploring interactions and implications.

The coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has presented global challenges with a diverse clinical spectrum, including severe respiratory complications and systemic effects. This review explores the intricate relationship between mitochondrial dysfunction, aging, and obesity in COVID-19. Mitochondria are vital for cellular energy provision and resilience against age-related macromolecule damage accumulation. They manage energy allocation in cells, activating adaptive responses and stress signals such as redox imbalance and innate immunity activation. As organisms age, mitochondrial function diminishes. Aging and obesity, linked to mitochondrial dysfunction, compromise the antiviral response, affecting the release of interferons, and worsening COVID-19 severity. Furthermore, the development of post-acute sequelae of SARS-CoV-2 infection (PASC), also known as long COVID has been associated with altered energy metabolism, and chronic immune dysregulation derived from mitochondrial dysfunction. Understanding the interplay between mitochondria, aging, obesity, and viral infections provides insights into COVID-19 pathogenesis. Targeting mitochondrial health may offer potential therapeutic strategies to mitigate severe outcomes and address long-term consequences in infected individuals.

Socioeconomic factors and COVID-19 mortality in immune-mediated rheumatic diseases patients: regional analysis from Argentina, Mexico and Brazil.

BACKGROUND: SARS-CoV-2 infection has become a major international issue, not only from a medical point of view, but also social, economic and political. Most of the available information comes from the United States, Europe, and China, where the population and the socioeconomic status are very different from Latin American countries. This study evaluates the effect of regional socioeconomic characteristics on mortality due SARS-CoV-2 infection in patients with immune-mediated rheumatic diseases (IMRD) from Argentina, Mexico and Brazil. METHODS: Data from three national registries, SAR-COVID (Argentina), CMR-COVID (Mexico) and ReumaCoV-Brasil (Brazil), were combined. Adult IMRD patients with SARS-CoV-2 infection were recruited. National data for each province/state, including population density, number of physicians per inhabitant, income, unemployment, GINI index, Municipal Human Development Index (MHDI), stringency index, vaccination rate and most frequent viral strains per period were assessed as risk factors for mortality due to COVID-19. RESULTS: A total of 4744 patients were included, 2534 (53.4%) from SAR-COVID, 1166 (24.6%) from CMRCOVID and 1044 (22.0%) from ReumaCoV-Brasil. Mortality due to COVID-19 was 5.4%. In the multivariable analysis, higher number of physicians per 1000 inhabitants and being infected during the vaccination period of each country were associated with lower mortality. After adjustment for socioeconomic factors, there was no association with country of residence and mortality. CONCLUSION: These findings corroborate the complex interplay between socioeconomic factors, rheumatic disease activity, and regional disparities as determinants of death due to COVID-19 in Argentina, Brazil and Mexico. Thus, this research provides valuable insights for guiding public health policies and clinical practice in the ongoing fight against the COVID-19 pandemic.

Inline Raman spectroscopy as process analytical technology for SARS-CoV-2 VLP production.

The present work focused on inline Raman spectroscopy monitoring of SARS-CoV-2 VLP production using two culture media by fitting chemometric models for biochemical parameters (viable cell density, cell viability, glucose, lactate, glutamine, glutamate, ammonium, and viral titer). For that purpose, linear, partial least square (PLS), and nonlinear approaches, artificial neural network (ANN), were used as correlation techniques to build the models for each variable. ANN approach resulted in better fitting for most parameters, except for viable cell density and glucose, whose PLS presented more suitable models. Both were statistically similar for ammonium. The mean absolute error of the best models, within the quantified value range for viable cell density (375,000-1,287,500 cell/mL), cell viability (29.76-100.00%), glucose (8.700-10.500 g/), lactate (0.019-0.400 g/L), glutamine (0.925-1.520 g/L), glutamate (0.552-1.610 g/L), viral titer (no virus quantified-7.505 log10 PFU/mL) and ammonium (0.0074-0.0478 g/L) were, respectively, 41,533 ± 45,273 cell/mL (PLS), 1.63 ± 1.54% (ANN), 0.058 ± 0.065 g/L (PLS), 0.007 ± 0.007 g/L (ANN), 0.007 ± 0.006 g/L (ANN), 0.006 ± 0.006 g/L (ANN), 0.211 ± 0.221 log10 PFU/mL (ANN), and 0.0026 ± 0.0026 g/L (PLS) or 0.0027 ± 0.0034 g/L (ANN). The correlation accuracy, errors, and best models obtained are in accord with studies, both online and offline approaches while using the same insect cell/baculovirus expression system or different cell host. Besides, the biochemical tracking throughout bioreactor runs using the models showed suitable profiles, even using two different culture media.

Impact of COVID-19 infection on genitourinary cancer management. SOGUG-COVID-19: A spanish, multicenter, observational study.

INTRODUCTION: The COVID-19 pandemic is a great burden worldwide, but its impact on patients with genitourinary cancer (GUC) is poorly characterized. This study aimed to characterize the clinical features and evolution of GUC patients affected by COVID-19 in Spain. PATIENTS AND METHODS: SOGUG-COVID-19 was an observational ambispective non-interventional study that recruited patients with SARS-CoV-2 infection who had been treated for GUC in 32 Spanish hospitals. Data were collected from patients' medical records in a short period of time, coinciding with the first waves of COVID-19, when the mortality was also higher in the general population. RESULTS: From November 2020 to April 2021, 408 patients were enrolled in the study. The median age was 70 years, and 357 patients (87.5%) were male. Most frequent Cancer Origin was: prostate (40.7%), urothelial (31.4%) and kidney (22.1%). Most patients (71.3%) were diagnosed at the metastatic stage, and 33.3% had poorly differentiated histology. Anticancer treatment during the infection was reported in 58.3% of patients, and 21.3% had received immunotherapy prior to or concurrent with the infection. The most frequent COVID-19 symptoms were pyrexia (49.0%), cough (38.2%) and dyspnea (31.9%). Median age was higher for patients with pneumonia (p < 0.001), patchy infiltrates (p = 0.005), ICU admission (p < 0.001) and death (p < 0.001). Tumor stage was associated with complications (p = 0.006). The fatality rate was 19.9% and the 6-month COVID-19-specific survival rate was 79.7%. CONCLUSION: Patients with genitourinary cancers seem exceptionally vulnerable to COVID-19 regardless of tumor type or anticancer therapy. Age and tumor stage were the only identified risk factors for severe COVID-19.

SpiN-Tec: A T cell-based recombinant vaccine that is safe, immunogenic, and shows high efficacy in experimental models challenged with SARS-CoV-2 variants of concern.

The emergence of new SARS-CoV-2 variants of concern associated with waning immunity induced by natural infection or vaccines currently in use suggests that the COVID-19 pandemic will become endemic. Investing in new booster vaccines using different platforms is a promising way to enhance protection and keep the disease under control. Here, we evaluated the immunogenicity, efficacy, and safety of the SpiN-Tec vaccine, based on a chimeric recombinant protein (SpiN) adjuvanted with CTVad1 (MF59-based adjuvant), aiming at boosting immunity against variants of concern of SARS-CoV-2. Immunization of K18-hACE-2 transgenic mice and hamsters induced high antibody titers and cellular immune response to the SpiN protein as well as to its components, RBD and N proteins. Importantly in a heterologous prime/boost protocol with a COVID-19 vaccine approved for emergency use (ChAdOx1), SpiN-Tec enhanced the level of circulation neutralizing antibodies (nAb). In addition to protection against the Wuhan isolate, protection against the Delta and Omicron variants was also observed as shown by reduced viral load and lung pathology. Toxicity and safety tests performed in rats demonstrated that the SpiN-Tec vaccine was safe and, based on these results, the SpiN-Tec phase I/II clinical trial was approved.

Massive pulmonary thromboembolism in an adolescent with SARS-CoV-2 infection. / Tromboembolismo pulmonar masivo en adolescente con infección por SARS­CoV-2.

Thromboembolic events incidence is low in pediatrics; high suspicion and explicit management algorithms are essential. We present a 12-year-old female patient with two weeks of dyspnea, orthopnea, and ankle edema. Tests showed metabolic acidosis, hyperlactatemia, elevated D-dimer, and positive SARS-CoV-2 CRP. The echocardiogram showed severe right ventricular dysfunction and supra-systemic pulmonary hypertension. Chest CT angiography showed extensive bilateral pulmonary thromboembolism. Anticoagulant therapy was started. She presented with hemodynamic instability. Adrenaline, norepinephrine, milrinone, and nitric oxide were started. The clinical picture was extremely severe in the first 24 hours. It was decided to perform systemic thrombolysis with alteplase, which led to an improvement. Cardiorespiratory stabilization and anticoagulation are the mainstays of therapy in massive pulmonary thromboembolism. Fibrinolytic therapy is used in selected high-risk cases. In this patient, systemic reperfusion therapy with alteplase was performed with no significant complications.

La incidencia de eventos tromboembólicos es baja en pediatría; es importante la alta sospecha y algoritmos claros de manejo. Presentamos una paciente de 12 años, con 2 semanas de disnea, ortopnea y edema de tobillos. Exámenes evidenciaron acidosis metabólica, hiperlactatemia, dímero D elevado y PCR SARS-CoV-2 positiva. Ecocardiograma con disfunción grave de ventrículo derecho e hipertensión pulmonar suprasistémica. Angio-TAC de tórax mostró extenso tromboembolismo pulmonar bilateral. Se inició terapia anticoagulante. Evolucionó con inestabilidad hemodinámica. Inició adrenalina, norepinefrina, milrinona y óxido nítrico. Primeras 24 horas de extrema gravedad. Se decidió realización de trombólisis sistémica con alteplasa, y evolucionó con progresión a la mejoría. Los pilares de terapia en tromboembolismo pulmonar (TEP) masivo son la estabilización cardiorrespiratoria y la anticoagulación. En casos seleccionados como de alto riesgo, se utiliza terapia fibrinolítica. En esta paciente, se realizó terapia de reperfusión sistémica con alteplasa, sin complicaciones de importancia en el procedimiento.

Acute kidney injury and mortality in patients with critical COVID-19 in Mexico: case-control study.

OBJECTIVE: We aimed to test the association between acute kidney injury (AKI) and mortality in critically ill patients with Coronavirus disease 2019 (COVID-19). METHOD: We conducted a single-center case-control study at the intensive care unit (ICU) of a second-level hospital in Mexico. We included 100 patients with critical COVID-19 from January to December 2021, and collected demographic characteristics, comorbidities, APACHE II, SOFA, NEWS2, and CO-RADS scores at admission, incidence of intrahospital complications, length of hospital and ICU stay, and duration of mechanical ventilation, among others. RESULTS: The median survival of deceased patients was 20 days. After multivariable logistic regression, the following variables were significantly associated to mortality: AKI (adjusted odds ratio [AOR] 6.64, 95% confidence intervals [CI] = 2.1-20.6, p = 0.001), age > 55 years (AOR 5.3, 95% CI = 1.5-18.1, p = 0.007), and arrhythmias (AOR 5.15, 95% CI = 1.3-19.2, p = 0.015). Median survival was shorter in patients with AKI (15 vs. 22 days, p = 0.043), as well as in patients with overweight/obesity (15 vs. 25 days, p = 0.026). CONCLUSION: Our findings show that the development of AKI was the main risk factor associated with mortality in critical COVID-19 patients, while other factors such as older age and cardiac arrhythmias were also associated with this outcome. The management of patients with COVID-19 should include renal function screening and staging on admission to the Emergency Department.

OBJETIVO: Probar la asociación entre lesión renal aguda y mortalidad en pacientes con COVID-19 grave. MÉTODO: Realizamos un estudio de casos y controles unicéntrico en la unidad de cuidados intensivos (UCI) de un hospital de segundo nivel en México. Incluimos 100 pacientes con COVID-19 grave de enero a diciembre 2021, recolectando características demográficas, comorbilidad, APACHE II, SOFA, NEWS2 y CO-RADS al ingreso, incidencia de complicaciones intrahospitalarias, duración de la estancia hospitalaria y en la UCI, duración de ventilación mecánica, etc. RESULTADOS: La mediana de supervivencia de los pacientes que fallecieron fue de 20 días. Al realizar el análisis de regresión logística multivariable, las siguientes variables se asociaron significativamente con la mortalidad: lesión renal aguda (odds ratio ajustada [ORa]: 6.64; intervalo de confianza del 95% [IC95%]: 2.1-20.6; p = 0.001), edad > 55 años (ORa: 5.3; IC95%: 1.5-18.1; p = 0.007) y arritmias (ORa: 5.15; IC95%: 1.3-19.2; p = 0.015). La supervivencia fue menor en pacientes con lesión renal aguda (15 vs. 22 días; p = 0,043), así como en pacientes con sobrepeso u obesidad (15 vs. 25 días; p = 0.026). CONCLUSIONES: Nuestros resultados muestran que el desarrollo de lesión renal aguda es el principal factor de riesgo asociado a mortalidad en pacientes con COVID-19 grave, mientras que otros factores, como la edad > 55 años y la presencia de arritmias cardiacas, también se asocian a mortalidad por COVID-19. El manejo de pacientes con COVID-19 debe incluir el tamizaje y la estadificación de la función renal al ingreso a urgencias.

Variables Associated With In-Hospital Lethality in COVID-19: A Prospective Cohort Study From Colombia.

INTRODUCTION: The emergence of COVID-19 represents the most significant health crisis in recent history. Incidence and mortality rates depend on several factors. Many studies have focused on investigating which characteristics could be strongly related to higher mortality and lethality. OBJECTIVE: This study aims to analyze the variables associated with in-hospital mortality among patients admitted in a reference northeastern region of a Colombian institution. METHODS: An ambidirectional cohort, single-center study was carried out in a reference hospital in northeastern Colombia. All patients admitted to the Fundación Oftalmológica de Santander (FOSCAL) between March 2020 and September 2021, with COVID-19 real-time polymerase chain reaction (PCR) positive test, were included. RESULTS: A total of 3,028 patients were included, of whom 2,034 (67.8%) survived and 994 (32.8%) died during their hospital stay; 48.8% (1,479) of the patients were female. The most common comorbidities were hypertension (1,236 patients, 40.8%), obesity (body mass index (BMI) ≥ 30; 656 patients, 21.6%), and diabetes (618 patients, 20.4%). The average age of the surviving patients was 52.2 years, while for the deceased patients, it was 70.3 years. The variables that showed significant association with in-hospital mortality were as follows: male sex ≥ 45 years, dyspnea, oxygen saturation (SatO2) < 85%, hypertension, chronic kidney disease (CKD), and a Charlson Comorbidity Index (CCI) score of >1. CONCLUSIONS: Male sex, age ≥ 45 years, dyspnea, SatO2 < 85%, hypertension, CKD, and a CCI score of >1 were associated with a higher risk of in-hospital mortality in COVID-19-infected patients.

Physiotherapeutic Approaches in the Rehabilitation of Patients After Severe Acute Respiratory Syndrome Coronavirus 2: A Scoping Review of In-Hospital Until Outpatient Phase.

OBJECTIVES: This scoping review aimed to identify studies that address the use of physical therapy in the rehabilitation of adult and elderly patients after SARS-CoV-2 infection. METHODS: This scoping review was based on Joanna Briggs Institute methodology: participant (P), represented by adult and elderly patients after SARS-CoV-2; concept (C), main physical therapy interventions; and context (C), recovery period after SARS-CoV-2 (in-hospital or outpatient phase). The following information sources were used: MEDLINE/PubMed, Cochrane Library, PEDro, Scopus, CINAHL, Web of Science, Science Direct, and Springer. The search was conducted between December 2019 and November 2021. Rayyan (Qatar Computing Research Institute, QCRI) was used for study selection process and analysis. RESULTS: A total of 7,568 studies were identified; 11 were included in this review. The most frequent physical therapy interventions were those associated with pulmonary, cardiac, musculoskeletal, neurological, and digestive rehabilitation in adult and elderly patients after SARS-CoV-2. Physiotherapy interventions included aerobic exercises, respiratory muscle training, muscle strength training, breathing exercises, early mobilization, balance training, bronchial hygiene maneuvers, body positioning management, flexibility training, cognitive training, neuromuscular electrical stimulation, and trunk exercises. CONCLUSION: Our study demonstrated the growing utility of therapeutic interventions, mainly in improving quality of life, as well as body functions after pulmonary, cardiac, neurological, digestive, and musculoskeletal rehabilitation in patients after SARS-CoV-2.

In Silico Design of miniACE2 Decoys with In Vitro Enhanced Neutralization Activity against SARS-CoV-2, Encompassing Omicron Subvariants.

The COVID-19 pandemic has overwhelmed healthcare systems and triggered global economic downturns. While vaccines have reduced the lethality rate of SARS-CoV-2 to 0.9% as of October 2024, the continuous evolution of variants remains a significant public health challenge. Next-generation medical therapies offer hope in addressing this threat, especially for immunocompromised individuals who experience prolonged infections and severe illnesses, contributing to viral evolution. These cases increase the risk of new variants emerging. This study explores miniACE2 decoys as a novel strategy to counteract SARS-CoV-2 variants. Using in silico design and molecular dynamics, blocking proteins (BPs) were developed with stronger binding affinity for the receptor-binding domain of multiple variants than naturally soluble human ACE2. The BPs were expressed in E. coli and tested in vitro, showing promising neutralizing effects. Notably, miniACE2 BP9 exhibited an average IC50 of 4.9 µg/mL across several variants, including the Wuhan strain, Mu, Omicron BA.1, and BA.2 This low IC50 demonstrates the potent neutralizing ability of BP9, indicating its efficacy at low concentrations.Based on these findings, BP9 has emerged as a promising therapeutic candidate for combating SARS-CoV-2 and its evolving variants, thereby positioning it as a potential emergency biopharmaceutical.

Waning immune response to CoronaVac in health care workers from different age groups.

Introduction: The waning of serum antibodies against severe acute respiratory syndrome coronavirus 2 observed in several studies raises questions about long-term immunity. Lower antibody levels are associated with new cases of COVID-19 even postvaccination, leading to the administration of booster doses. Objectives: To evaluate the postvaccination immune humoral response and the relationship between postvaccination seropositivity rates and demographic data among health care workers 6 months after CoronaVac vaccination. Methods: This was a cross-sectional study including health care workers vaccinated with two doses of CoronaVac after 6 months or more. The present study was conducted with the analysis of postvaccination serology test to assess the level of humoral response (anti-receptor binding domain IgG) after vaccination. Results: A total of 325 participants were enrolled, of whom 76% were female, with a median age of 42 years (20-85; interquartile range 31-53). Overall, 18.8% (61) of the participants results were seropositive for anti-receptor binding domain IgG; 81.2% did not have sufficient quantitative titers. The IgG titers obtained from female health care workers did not differ from those obtained from seropositive male health care workers, regardless of age. Conclusions: A group of positive quantitative titers was identified in the serology test for IgG antibodies against severe acute respiratory syndrome coronavirus 2. Further studies are needed to determine the durability of postvaccination antibodies and how serology testing can be used to determine the ideal timing for booster doses of the vaccine.

Introdução: O declínio dos anticorpos séricos contra a síndrome respiratória aguda grave do coronavirus 2 observado em vários estudos levanta questões sobre a imunidade a longo prazo. Níveis mais baixos de anticorpos estão associados a novos casos de covid-19 mesmo após a vacinação, levando à administração de doses de reforço. Objetivos: Avaliar a resposta imunitária humoral após a vacinação e a relação entre as taxas de soropositividade após a vacinação e dados demográficos em trabalhadores da saúde por mais de 6 meses após a imunização com CoronaVac. Métodos: Estudo transversal incluindo profissionais de saúde vacinados com duas doses de CoronaVac após 6 meses ou mais. O estudo foi realizado com a análise do teste sorológico após a vacinação para avaliar os níveis de resposta humoral (anti-domínio de ligação ao receptor IgG) após a vacinação. Resultados: Neste estudo foram incluídos 325 participantes, 76% do sexo feminino e a idade mediana foi de 42 anos (20-85; intervalo interquartil 31-53). No geral, 18,8% (61) dos resultados dos participantes foram soropositivos para IgG anti-domínio de ligação ao receptor; 81,2% não apresentaram títulos quantitativos suficientes. Os títulos de IgG obtidos para os profissionais de saúde do sexo feminino não foram diferentes daqueles obtidos para os participantes do sexo masculino com soropositividade, independentemente da idade. Conclusões: Foi possível identificar um grupo com títulos quantitativos positivos no teste sorológico para anticorpo IgG contra a síndrome respiratória aguda grave do coronavírus 2. Mais investigações são necessárias para determinar a durabilidade dos anticorpos após a vacinação e como os testes sorológicos podem determinar o momento ideal das doses de reforço da vacina.

Detection of Neutralizing Antibodies in Serum Samples Using a SARS-CoV-2 Pseudotyped Virus Assay.

Conventional live virus research on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causal agent of coronavirus disease-19 (COVID-19), requires Biosafety Level 3 (BSL-3) facilities. SARS-CoV-2 pseudotyped viruses have emerged as valuable tools in virology, mimicking the entry process of the SARS-CoV-2 virus into human cells by expressing its spike glycoprotein in a surrogate system using recombinant plasmids. One significant application of this tool is in functional assays for the evaluation of neutralizing antibodies. Pseudotyped viruses have the advantage of being competent for only a single cycle of infection, providing better safety and versatility and allowing them to be studied in BSL-2 laboratories. Here, we describe three protocols for the detection of SARS-CoV-2 neutralizing antibodies through a pseudotyped virus assay. First, SARS-CoV-2 S pseudotyped viruses (PV SARS-CoV-2 S) are produced using a Moloney murine leukemia virus (MuLV) three-plasmid system. The plasmids are designed to express the GagPol packing proteins, enhanced green fluorescent protein (eGFP) as a readout system, and the SARS-CoV-2 S protein modified to remove the endoplasmic reticulum retention domain and to improve infection. Next, the internalization of PV SARS-CoV-2 S protein in human embryonic kidney 293T (HEK-293T) cells overexpressing angiotensin-converting enzyme 2 (HEK-293T-ACE2) is confirmed by fluorescence microscopy and quantified using flow cytometry. Finally, PV SARS-CoV-2 S is used to screen neutralizing antibodies in serum samples from convalescent COVID-19 patients; it can also be used for studying the cell entry mechanisms of different SARS-CoV-2 variants, evaluating antiviral agents, and designing vaccines. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Generation of PV SARS-CoV-2 S pseudotyped virus Basic Protocol 2: Assay of PV SARS-CoV-2 S internalization in target cells. Basic Protocol 3: Detection of neutralizing antibodies in serum samples.

Complete Blood Count in Children With COVID-19: A Predictor of Disease Severity.

Blood count abnormalities are frequent in patients with severe COVID-19 disease and there is still a lack of information in pediatric complete blood count (CBC) results. Thus, this study aims to correlate the CBC in the emergency room of children with COVID-19 between 0 and 10 years old and the clinical severity of the disease. A retrospective cohort study was performed in children with COVID-19 who collected at the emergency room CBC, C-reactive protein (CRP), platelet to lymphocyte ratio (PLR), neutrophil to lymphocyte ratio (NLR), neutrophil to monocyte ratio (NMR), lymphocyte to neutrophil ratio (LNR), lymphocyte to monocyte ratio (LMR), monocyte to neutrophil ratio (MNR) and monocyte to lymphocyte ratio (MLR). In total, demographic data from 93 children with median age of 19 months (0.3-126), 60.2% males, were included. The main changes in the CBC were atypical lymphocytes (51.6%) and eosinopenia (49.5%). From 69 hospitalized children, 21 were considered severe. There was no association between age, gender, and CRP value with clinical severity. The presence of underlying disease was five times higher (odds ratio [OR] = 5.08) in patients who required hospitalization and a higher NLR value was 54% (OR = 1.54) more likely to occur. Eosinopenia was three times more frequent in inpatients with disease severity criteria (OR = 3.05). In conclusion, children younger than 10 years of age with COVID-19 have changes in the CBC collected in the emergency room, mainly atypical lymphocytes and eosinopenia. The presence of a comorbidity or a higher NLR increases the chance of hospitalization. In addition, eosinopenia was a predictor of severity in inpatient children due to COVID-19.

An integrative analysis of SARS-CoV 2 during the first and second waves of COVID-19 in Salta, Argentina.

Wastewater surveillance has been extensively applied to provide information about SARS-CoV-2 circulation in the community. However, its applicability is limited in regions lacking adequate sewerage infrastructure, without wastewater treatment plants (WWTP) or with insufficient coverage. During the COVID-19 pandemic, from July 2020 to September 2021, comprehensive epidemiological data encompassing positive, recovered, and deceased cases were collected alongside precipitation records. Additionally, wastewater samples from 13 main sewersheds and river water from two points (up- and downstream the main WWTP), in the city of Salta, were gathered. A total of 452 water samples were analyzed for quantitative detection of SARS-CoV-2 using reverse transcription real-time PCR. Across the 62-week study period, two distinct waves of COVID-19 were identified. The dynamics of deceased cases showed peaks 10 and 28 days after the peaks of positive cases in the first and second waves, respectively. Downstream river water exhibited higher fecal contamination than the upstream samples, evincing the impact of the WWTP discharges. Viral concentration in river waters mirrored those from wastewater, reflecting the progression of cases. Despite the lower reported number of cases during the first wave in comparison to the second (5420 vs. 8516 cases at the respective peaks), higher viral concentrations were detected in water samples (1.97 × 107 vs. 2.36 × 106 gc/L, respectively), suggesting underreporting during the first wave, and highlighting the positive effect of vaccination during the second. To the best of our knowledge, this is the first study that simultaneously and systematically analyzed surface water and wastewater over a prolonged period, the effect of precipitations were considered for the variations in the concentrations, and the findings compared with epidemiological information. Environmental surveillance was demonstrated to be a great tool to obtain valuable information about the circulation patterns of SARS-CoV-2, especially under resource constraints to massively test the population, thus, underreporting cases. Furthermore, the methodology employed herein can be easily expanded to the community-level surveillance of other pathogens excreted in urine and feces, encompassing viruses, bacteria, and protozoa.

The Model for End-Stage Liver Disease (MELD) Score Predicting Mortality Due to SARS-CoV-2 in Mexican Patients.

Background/Objectives: Coronavirus Disease 2019 (COVID-19) can cause liver injury and a deterioration of hepatic function. The Model for End-Stage Liver Disease (MELD) score is a good predictor for poor prognosis of hospitalized COVID-19 patients in the United States, Egypt and Turkey. Nevertheless, the best cut-off value for the MELD score to predict mortality in the Mexican population has yet to be established. Methods: A total of 234 patients with COVID-19 were studied in a tertiary-level hospital. Patients were stratified into survivors (n = 139) and non-survivors (n = 95). Receiver operating characteristic curves, Cox proportional hazard models, Kaplan-Meier method, and Bonferroni corrections were performed to identify the predictors of COVID-19 mortality. Results: MELD score had an area under the curve of 0.62 (95% CI: 0.56-0.68; p = 0.0009), sensitivity = 53.68%, and specificity = 73.38%. Univariate Cox proportional hazard regression analysis suggested that the leukocytes > 10.6, neutrophils > 8.42, neutrophil-to-lymphocyte ratio (NLR) > 8.69, systemic immune-inflammation index (SII) > 1809.21, MELD score > 9, and leukocyte glucose index (LGI) > 2.41 were predictors for mortality. However, the multivariate Cox proportional hazard model revealed that only the MELD score >9 (Hazard Ratio [HR] = 1.83; 95% confidence interval [CI]: 1.2-2.8; Pcorrected = 0.03) was an independent predictor for mortality of COVID-19. Conclusions: Although the MELD score is used for liver transplantation, we suggest that a MELD score >9 could be an accurate predictor for COVID-19 mortality at admission to ICU requiring mechanical ventilation.

Prospective Variation of Cytokine Trends during COVID-19: A Progressive Approach from Disease Onset until Outcome.

COVID-19 is characterized by pronounced hypercytokinemia. The cytokine switch, marked by an imbalance between pro-inflammatory and anti-inflammatory cytokines, emerged as a focal point of investigation throughout the COVID-19 pandemic. However, the kinetics and temporal dynamics of cytokine release remain contradictory, making the development of new therapeutics difficult, especially in severe cases. This study collected serum samples from SARS-CoV-2 infected patients at 72 h intervals and monitored them for various cytokines at each timepoint until hospital discharge or death. Cytokine levels were analyzed based on time since symptom onset and patient outcomes. All cytokines studied prospectively were strong predictors of mortality, particularly IL-4 (AUC = 0.98) and IL-1ß (AUC = 0.96). First-timepoint evaluations showed elevated cytokine levels in the mortality group (p < 0.001). Interestingly, IFN-γ levels decreased over time in the death group but increased in the survival group. Patients who died exhibited sustained levels of IL-1ß and IL-4 and increased IL-6 levels over time. These findings suggest cytokine elevation is crucial in predicting COVID-19 mortality. The dynamic interplay between IFN-γ and IL-4 highlights the balance between Th1/Th2 immune responses and underscores IFN-γ as a powerful indicator of immune dysregulation throughout the infection.

Inter-institutional laboratory standardization for SARS-CoV-2 surveillance through wastewater-based epidemiology applied to Mexico City.

Objectives: Wastewater-based surveillance applied to SARS-CoV-2 viral load quantification for COVID-19 has become one of the most relevant complementary tools in epidemiologic prevention programs worldwide. However, this valuable decision-making tool still requires fine-tuning to produce comparable results between laboratories, especially when applied to the surveillance of megacities. Methods: Six laboratories across Mexico and one from the United States executed an interlaboratory study to set up a singular standardized protocol considering method cost, installed infrastructure, materials available, and supply availability for SARS-CoV-2 quantification from five Mexico City sampling sites across this megacity. Results: Comparable data from processing outcomes in the Mexican laboratories and in the external international laboratory serve as a validating data source. The Bland-Altman comparison showed consistency, with cycle threshold values within ±1.96 SD of SARS-CoV-2 genetic copies for the standard curve quantification, with a mismatch of two laboratories. In addition, MS2 bacteriophage recovery rates varied between 35% and 67% among all participating laboratories. Finally, the efficiency of viral genetic material recovered from all participating laboratories varied between 65% and 93% for the participating laboratories. Conclusion: This work lays the foundation for extensive and continuous wastewater-based surveillance application across independent Mexican laboratories in a time- and resource-effective manner.