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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.
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COVID-19 , Citocinas , Interleucina-4 , SARS-CoV-2 , Humanos , COVID-19/inmunología , COVID-19/sangre , COVID-19/mortalidad , Masculino , Femenino , Persona de Mediana Edad , Citocinas/sangre , SARS-CoV-2/inmunología , Anciano , Interleucina-4/sangre , Estudios Prospectivos , Interferón gamma/sangre , Interleucina-1beta/sangre , Adulto , Interleucina-6/sangreRESUMEN
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.
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Anticuerpos Neutralizantes , Anticuerpos Antivirales , COVID-19 , SARS-CoV-2 , Humanos , Anticuerpos Neutralizantes/inmunología , Anticuerpos Neutralizantes/sangre , SARS-CoV-2/inmunología , Anticuerpos Antivirales/inmunología , Anticuerpos Antivirales/sangre , COVID-19/virología , COVID-19/inmunología , COVID-19/diagnóstico , COVID-19/sangre , Pruebas de Neutralización/métodos , Células HEK293 , Pseudotipado Viral , Glicoproteína de la Espiga del Coronavirus/inmunología , Glicoproteína de la Espiga del Coronavirus/genéticaRESUMEN
OBJECTIVE: The study aimed to determine the association between serum magnesium and Vitamin D levels with the severity and mortality by coronavirus disease 19 (COVID-19) in hospitalized patients. METHOD: Men and women over 18 years of age with probable COVID-19 were enrolled in a case-control study. Patients with a positive or negative test for Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were allocated into case or control groups, respectively. Vitamin D deficiency was defined by concentrations < 20 ng/mL and hypomagnesemia by serum levels < 1.8 mg/dL. RESULTS: A total of 54 patients, 30 women and 24 men, were enrolled and allocated into the groups with (n = 27) and without (n = 27) COVID-19. The logistic regression analysis showed that Vitamin D deficiency (odds ratio [OR] = 6.13; 95% confidence intervals [CI]: 1.32-28.34) and insufficiency (OR = 0.12; 95% CI: 0.02-0.60) are significantly associated with hospitalization. However, Vitamin D disorders and hypomagnesemia were not associated with mortality. CONCLUSIONS: The results of the present study revealed that Vitamin D disturbances, but not hypomagnesemia, are associated with the severity of SARS-CoV-2.
OBJETIVO: Determinar la asociación entre los niveles séricos de vitamina D y de magnesio con la gravedad y la mortalidad de la COVID-19 en pacientes hospitalizados. MÉTODO: Hombres y mujeres mayores de 18 años con probable COVID-19 fueron enrolados en un estudio de casos y controles. Los pacientes con una prueba positiva o negativa para SARS-CoV-2 fueron asignados en los grupos de casos y de controles, respectivamente. RESULTADOS: Un total de 54 pacientes, 30 mujeres y 24 hombres, fueron enrolados y asignados a los grupos COVID-19 (n = 27) y control (n = 27). El análisis de regresión logística mostró que la deficiencia de vitamina D (odds ratio [OR]: 6.13; intervalo de confianza del 95% [IC95%]: 1.32-28.34) y la insuficiencia de vitamina D (OR: 0.12; IC95%: 0.02-0.60) se asocian significativamente con hospitalización. Sin embargo, las alteraciones de la vitamina D y la hipomagnesemia no se asociaron con mortalidad. CONCLUSIONES: Los resultados del presente estudio revelaron que las alteraciones de la vitamina D, pero no la hipomagnesemia, se asocian con la gravedad de la COVID-19.
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COVID-19 , Deficiencia de Magnesio , Magnesio , Índice de Severidad de la Enfermedad , Deficiencia de Vitamina D , Vitamina D , Humanos , COVID-19/sangre , COVID-19/mortalidad , COVID-19/complicaciones , Masculino , Femenino , Magnesio/sangre , Persona de Mediana Edad , Estudios de Casos y Controles , Deficiencia de Vitamina D/sangre , Deficiencia de Vitamina D/complicaciones , Deficiencia de Vitamina D/epidemiología , Vitamina D/sangre , Vitamina D/análogos & derivados , Anciano , Deficiencia de Magnesio/sangre , Deficiencia de Magnesio/complicaciones , Deficiencia de Magnesio/epidemiología , Adulto , Hospitalización/estadística & datos numéricos , SARS-CoV-2RESUMEN
COVID-19 is characterized by a broad range of symptoms and disease trajectories. Understanding the correlation between clinical biomarkers and lung pathology during acute COVID-19 is necessary to understand its diverse pathogenesis and inform more effective treatments. Here, we present an integrated analysis of longitudinal clinical parameters, peripheral blood markers, and lung pathology in 142 Brazilian patients hospitalized with COVID-19. We identified core clinical and peripheral blood signatures differentiating disease progression between patients who recovered from severe disease compared with those who succumbed to the disease. Signatures were heterogeneous among fatal cases yet clustered into two patient groups: "early death" (<15 days until death) and "late death" (>15 days). Progression to early death was characterized systemically and in lung histopathological samples by rapid endothelial and myeloid activation and the presence of thrombi associated with SARS-CoV-2+ macrophages. In contrast, progression to late death was associated with fibrosis, apoptosis, and SARS-CoV-2+ epithelial cells in postmortem lung tissue. In late death cases, cytotoxicity, interferon, and T helper 17 (TH17) signatures were only detectable in the peripheral blood after 2 weeks of hospitalization. Progression to recovery was associated with higher lymphocyte counts, TH2 responses, and anti-inflammatory-mediated responses. By integrating antemortem longitudinal blood signatures and spatial single-cell lung signatures from postmortem lung samples, we defined clinical parameters that could be used to help predict COVID-19 outcomes.
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COVID-19 , Progresión de la Enfermedad , Pulmón , SARS-CoV-2 , Humanos , COVID-19/sangre , COVID-19/diagnóstico , Pulmón/patología , SARS-CoV-2/aislamiento & purificación , Masculino , Femenino , Persona de Mediana Edad , Biomarcadores/sangre , Análisis de la Célula Individual , Adulto , Brasil , AncianoRESUMEN
Introduction: COVID-19 is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a virus notable for its rapid mutation rate, which has led to the emergence of various variants such as Delta and Omicron, each with potentially different levels of transmissibility and virulence. Therefore, this study aims to compare clinical charactheristics and markers associated with the severity of COVID-19 in hospitalized patients from western Mexico who were infected with the Delta and Omicron variants of SARS-CoV-2. Methods: This cross-sectional study involved 66 patients hospitalized for COVID-19, diagnosed by RT-qPCR. SARS-CoV-2 variants were identified through whole genome sequencing using the COVIDseq platform from Illumina. Upon admission, patients underwent a clinical history assessment, blood gas analysis, and blood biometry. Additionally, several tests and markers were measured, including the percentage of neutralizing antibodies, erythrocyte sedimentation rate (ESR), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNFα), D-dimer, lactate dehydrogenase (LDH), C-reactive protein (CRP), and ferritin. Results and discussion: Patients hospitalized with the Omicron were found to be older, compared to those infected with the Delta (64 vs. 54 years, p = 0.006). Additionally, a higher proportion of male patients were observed in the Omicron compared to the Delta (p = 0.029). Both Omicron and Delta variants were associated with lymphopenia, although the lymphocyte count was lower in Omicron (0.9 vs. 0.56 10x3/L; p = 0.007). The COVID-GRAM scale indicated a high risk for severe disease in both groups, but the score was higher in Omicron compared to Delta (157 vs. 128 points; p = 0.0004). Patients infected with Omicron exhibited a lower percentage of neutralizing antibodies than those with Delta (35.99 vs. 81%; p < 0.05), regardless of their vaccination status. Among the markers assessed, globular ESR was found to be lower in Omicron compared to Delta (30.5 vs. 41.5 mm/h; p = 0.001), while ferritin levels were higher in patients infected with the Omicron (1,359 vs. 960.6 µg/L; p = 0.007). In patients with severe COVID-19, markers such as lymphopenia, neutralizing antibody levels, ferritin, and COVID-GRAM scores are elevated in the Omicron variant, while only the leukocyte count and ESR for the Delta variant.
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Biomarcadores , COVID-19 , Hospitalización , SARS-CoV-2 , Índice de Severidad de la Enfermedad , Humanos , COVID-19/diagnóstico , COVID-19/sangre , Masculino , México , Femenino , SARS-CoV-2/genética , Persona de Mediana Edad , Estudios Transversales , Biomarcadores/sangre , Adulto , AncianoRESUMEN
SARS-CoV-2 caused the pandemic situation experienced since the beginning of 2020, and many countries faced the rapid spread and severe form of the disease. Mechanisms of interaction between the virus and the host were observed during acute phase, but few data are available when related to immunity dynamics in convalescents. We conducted a longitudinal study, with 51 healthy donors and 62 COVID-19 convalescent patients, which these had a 2-month follow-up after symptoms recovery. Venous blood sample was obtained from all participants to measure blood count, subpopulations of monocytes, lymphocytes, natural killer cells and dendritic cells. Serum was used to measure cytokines, chemokines, growth factors, anti-N IgG and anti-S IgG/IgM antibodies. Statistic was performed by Kruskal-Wallis test, and linear regression with days post symptoms and antibody titers. All analysis had confidence interval of 95%. Less than 35% of convalescents were anti-S IgM+, while more than 80% were IgG+ in D30. Anti-N IgG decreased along time, with loss of seroreactivity of 13%. Eosinophil count played a distinct role on both antibodies during all study, and the convalescence was orchestrated by higher neutrophil-to-lymphocyte ratio and IL-15, but initial stages were marked by increase in myeloid DCs, B1 lymphocytes, inflammatory and patrolling monocytes, G-CSF and IL-2. Later convalescence seemed to change to cytotoxicity mediated by T lymphocytes, plasmacytoid DCs, VEGF, IL-9 and CXCL10. Anti-S IgG antibodies showed the longest perseverance and may be a better option for diagnosis. The inflammatory pattern is yet present on initial stage of convalescence, but quickly shifts to a reparative dynamic. Meanwhile eosinophils seem to play a role on anti-N levels in convalescence, although may not be the major causative agent. We must highlight the importance of immunological markers on acute clinical outcomes, but their comprehension to potentialize adaptive system must be explored to improve immunizations and further preventive policies.
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Anticuerpos Antivirales , COVID-19 , Convalecencia , Citocinas , Inmunoglobulina G , SARS-CoV-2 , Humanos , COVID-19/inmunología , COVID-19/sangre , Masculino , Femenino , Adulto , Persona de Mediana Edad , SARS-CoV-2/inmunología , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Citocinas/sangre , Inmunoglobulina M/sangre , Inmunoglobulina M/inmunología , Estudios Longitudinales , Anciano , Eosinófilos/inmunología , Eosinófilos/metabolismoRESUMEN
COVID-19's long-lasting and complex impacts have become a global concern, with diverse clinical outcomes. This study evaluated 226 participants to understand the clinical spectrum of COVID-19/Long COVID (LC), exploring how disease severity correlates with sociodemographic factors and biomarkers. Determinants related to COVID-19 severity included age (P < 0.001), lower education (P < 0.001), ethnicity (P = 0.003), overweight (P < 0.001), MTHFR gene rs1801133 (P = 0.035), cardiovascular diseases (P = 0.002), diabetes mellitus (DM) (P = 0.006), Factor VIII (FVIII) (P = 0.046), von Willebrand factor (VWF) (P = 0.002), and dimer D (DD) (P < 0.001). Six months later, in a portion of the monitored participants, a significant reduction in FVIII (P < 0.001), VWF (P = 0.002), and DD (P < 0.001) levels was observed, with only DD returning to normal values. Different systemic sequelae were identified, with higher incidences of joint pain and myalgia in participants with a clinical history of DM, chronic lung disease (CLD) and sustained high interleukin 6 values in the convalescent phase. CLD, COVID-19 severity and high DD levels increased the risk of developing dyspnea and palpitations. Women were more likely to develop lower limb phlebitis long-term, while sustained elevated FVIII in the convalescent phase was associated with an increased risk of swelling. Regular physical activity had a protective effect against swelling. This study highlights factors contributing to COVID-19 severity/LC, emphasizing endothelial cell activation as a potential mechanism.
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Biomarcadores , COVID-19 , SARS-CoV-2 , Índice de Severidad de la Enfermedad , Factor de von Willebrand , Humanos , COVID-19/sangre , Femenino , Masculino , Biomarcadores/sangre , Persona de Mediana Edad , Pronóstico , Factor de von Willebrand/metabolismo , Factor de von Willebrand/análisis , Adulto , Anciano , SARS-CoV-2/aislamiento & purificación , Factor VIII/metabolismo , Factor VIII/análisis , Productos de Degradación de Fibrina-Fibrinógeno/análisis , Productos de Degradación de Fibrina-Fibrinógeno/metabolismoRESUMEN
BACKGROUND: Pentraxin 3 (PTX3) is an acute-phase protein that belongs to the pentraxin family, which plays an important role in the body's defense against pathogens. PTX3 levels have been associated with inflammatory processes, and it is a possible biomarker for the diagnosis and prognosis of different infectious diseases, including COVID-19. The objective of this study was to analyze the potential of PTX3 as a plasma biomarker for predicting death in patients hospitalized with COVID-19. METHODS: The study included a total of 312 patients with COVID-19, admitted from July 2020 to August 2021 to hospital ward and intensive care unit beds at two hospitals in the Northeast Region of Brazil. PTX3 was measured using ELISA in samples collected within 24 h after hospital admission. Maximally selected rank statistics were used to determine the PTX3 cutoff point that best distinguished patients who died from those who survived. A receiver operating characteristic (ROC) curve was used to determine the performance of the biomarker. Survival analysis was performed using a Kaplan-Meier curve, and a Cox regression model was used to determine predictors associated with death. RESULTS: Of the 312 patients included in the study, 233 recovered and 79 died. Patients who died had higher PTX3 levels at the time of admission, when compared to those who recovered (median: 52.84 versus 10.79 ng/mL; p < 0.001). PTX3 showed area under the ROC (AUC) = 0.834, higher than other markers used in clinical practice, such as C-reactive protein (AUC = 0.72) and D-dimer (AUC = 0.77). Furthermore, according to the Kaplan-Meier survival curve, patients with PTX3 concentrations above the cutoff point (27.3 ng/mL) had a lower survival rate (p = 0.014). In multivariate Cox regression, PTX3 > 27.3 ng/mL was an important predictor of death, regardless of other confounding factors (hazard ratio = 1.79; p = 0.027). CONCLUSION: PTX3 can be considered as a potential biomarker for predicting death in patients hospitalized with COVID-19.
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Biomarcadores , Proteína C-Reactiva , COVID-19 , Hospitalización , Curva ROC , Componente Amiloide P Sérico , Humanos , Proteína C-Reactiva/análisis , Componente Amiloide P Sérico/análisis , Componente Amiloide P Sérico/metabolismo , COVID-19/mortalidad , COVID-19/diagnóstico , COVID-19/sangre , Masculino , Femenino , Biomarcadores/sangre , Persona de Mediana Edad , Anciano , Brasil/epidemiología , Pronóstico , SARS-CoV-2 , Adulto , Anciano de 80 o más AñosRESUMEN
Objectives: This study was performed to identify predictive markers of worse outcomes in patients with severe COVID-19 in an intensive care unit. Methods: Sixty patients with severe COVID-19, hospitalized in the Intensive Care Unit (ICU) between March and July 2021, were stratified into two groups according to the outcome survivors and non-survivors. After admission to the ICU, blood samples were collected directly for biomarker analysis. Routine hematological and biochemical biomarkers, as well as serum levels of cytokines, chemokines, and immunoglobulins, were investigated. Results: Lymphopenia, neutrophilia, and thrombocytopenia were more pronounced in non-surviving patients, while the levels of CRP, AST, creatinine, ferritin, AST, troponin I, urea, magnesium, and potassium were higher in the non-surviving group than the survival group. In addition, serum levels of IL-10, CCL2, CXCL9, and CXCL10 were significantly increased in patients who did not survive. These changes in the biomarkers evaluated were associated with increased mortality in patients with severe COVID-19. Conclusion: The present study confirmed and expanded the validity of laboratory biomarkers as indicators of mortality in severe COVID-19.
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Biomarcadores , COVID-19 , Unidades de Cuidados Intensivos , SARS-CoV-2 , Humanos , COVID-19/mortalidad , COVID-19/sangre , COVID-19/inmunología , Masculino , Biomarcadores/sangre , Femenino , Persona de Mediana Edad , Anciano , Citocinas/sangre , Hospitalización , Índice de Severidad de la Enfermedad , Pronóstico , Adulto , Anciano de 80 o más AñosRESUMEN
BACKGROUND: Compare the changes and differences in metabolome and lipidome profiles among severe COVID-19 and CAP patients with ARF to identify biomarkers that could be used for personalized diagnosis, prognosis, and treatment. RESEARCH DESIGN AND METHODS: Plasma samples were taken at hospital admission (baseline) and on the 5th day of hospitalization (follow-up) and examined by RP-LC-QTOF-MS and HILIC-LC-QTOF-MS. RESULTS: 127 patients, 17 with CAP and 110 with COVID-19, were included. The analysis revealed 87 altered metabolites, suggesting changes in the metabolism of arachidonic acid, glycerolipids, glycerophospholipids, linoleic acid, pyruvate, glycolysis, among others. Most of these metabolites are involved in inflammatory, hypoxic, and thrombotic processes. At baseline, the greatest differences were found in phosphatidylcholine (PC) 31:4 (p < 0.001), phosphoserine (PS) 34:3 (p < 0.001), and phosphatidylcholine (PC) 36:5 (p < 0.001), all of which were notably decreased in COVID-19 patients. At follow-up, the most dysregulated metabolites were monomethyl-phosphatidylethanolamine (PE-Nme) 40:5 (p < 0.001) and phosphatidylcholine (PC) 38:4 (p < 0.001). CONCLUSIONS: Metabolic and lipidic alterations suggest inhibition of innate anti-inflammatory and anti-thrombotic mechanisms in COVID-19 patients, which might lead to increased viral proliferation, uncontrolled inflammation, and thrombi formation. Results provide novel targets for predictive biomarkers against CAP and COVID-19. TRIAL REGISTRATION: Not applicable.
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Biomarcadores , COVID-19 , Infecciones Comunitarias Adquiridas , Humanos , COVID-19/sangre , Masculino , Femenino , Persona de Mediana Edad , Pronóstico , Biomarcadores/sangre , Anciano , Infecciones Comunitarias Adquiridas/sangre , Infecciones Comunitarias Adquiridas/diagnóstico , Lipidómica , Índice de Severidad de la Enfermedad , SARS-CoV-2 , Adulto , MetabolomaRESUMEN
Rapid virus identification is crucial for preventing outbreaks. The COVID-19 pandemic has highlighted the critical nature of rapid virus detection. Here, we designed a label-free electrochemical biosensor modified with gold nanoparticles (AuNPs) to detect IgG antibodies from human serum, enabling rapid point-of-care diagnostics. AuNPs were synthesized and characterized. A multivariate optimization was carried out to determine the optimal condition for functionalizing AuNPs with anti-IgG. Subsequently, using a glassy carbon electrode (GCE), a modified AuNPs/GCE electrochemical biosensor was developed for IgG detection. The results indicated that AuNPs displayed a spherical morphology with a size distribution of 19.54 nm. Additionally, the zeta potential was recorded at -7.84 mV. Central composite design (CCD) analysis determined the optimal conditions for functionalizing AuNPs to be an anti-IgG concentration of 320 µg mL-1, a temperature of 25 °C, and pH of 7.4. The characterization study confirmed the successful synthesis and functionalization of AuNPs. Through electrochemical impedance spectroscopy measurement, the biosensor demonstrated a limit of detection (LOD) of 0.2 ng mL-1 and limit of quantification (LOQ) of 0.8 ng mL-1. Furthermore, tests in real samples showed the interaction between IgG antibodies in serum samples and AuNPs/GCE, confirming the biosensor's ability to detect and quantify IgG in clinical samples.
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Técnicas Biosensibles , Técnicas Electroquímicas , Oro , Inmunoglobulina G , Límite de Detección , Nanopartículas del Metal , SARS-CoV-2 , Humanos , Técnicas Biosensibles/métodos , Oro/química , Nanopartículas del Metal/química , Técnicas Electroquímicas/métodos , Inmunoglobulina G/sangre , SARS-CoV-2/inmunología , SARS-CoV-2/aislamiento & purificación , COVID-19/diagnóstico , COVID-19/sangre , COVID-19/virología , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , ElectrodosRESUMEN
Acute respiratory distress syndrome is a significant complication in critical care patients. COVID-19 (C19)-associated severe respiratory failure is related to it, and d-dimer rise predicts a worse outcome. To investigate the association between d-dimer and the severity of this respiratory syndrome, we conducted a study in C19 intubated patients. A retrospective, single-center observational study was conducted with 64 C19 adult intubated patients. Strata of d-dimer results between patients was evaluated using survival analysis. Survival was higher in mild respiratory distress patients. D-dimer showed poor sensitivity and specificity in predicting respiratory failure severity. Risk assessment for death showed a higher prevalence of admission d-dimer results (HR 1.335; 95% CI 0.695-2.564). Our sample confidently represented the medical profile of C19 severe patients. Sepsis development in C19 is associated with the inflammatory storm in respiratory distress syndrome. As the receiver operating curves show, the increase in d-dimer results is consistent with inflammation rather than a prognostic biomarker. As expected, severe respiratory distress patients presented higher mortality. In summary, d-dimer results are not associated with the prognosis of C19 respiratory distress syndrome patients.
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Biomarcadores , COVID-19 , Productos de Degradación de Fibrina-Fibrinógeno , Síndrome de Dificultad Respiratoria , Humanos , Productos de Degradación de Fibrina-Fibrinógeno/análisis , COVID-19/sangre , COVID-19/complicaciones , COVID-19/mortalidad , Masculino , Biomarcadores/sangre , Estudios Retrospectivos , Síndrome de Dificultad Respiratoria/sangre , Síndrome de Dificultad Respiratoria/mortalidad , Femenino , Persona de Mediana Edad , Índice de Severidad de la Enfermedad , Anciano , Pronóstico , SARS-CoV-2 , Adulto , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: COVID-19 is a systemic infection with a significant impact on nutrition risk and the hematopoietic system. The neutrophil-lymphocyte ratio (NLR) may have prognostic value in determining severe cases of COVID-19 and the urea-creatinine ratio (UCR) is currently being studied as a potential biomarker of catabolism associated with critical illness. The aim was to assess the association between the NLR, UCR and C-reactive protein (CRP) with nutritional risk in hospitalized patients with COVID-19. METHODS: This is a retrospective cross-sectional study that assessed 589 hospitalized patients with COVID-19, 18 years of age or older, of both sexes. Nutritional risk was assessed by Nutritional Risk Screening (NRS, 2002) and NLR by neutrophils divided by lymphocyte count. The UCR was calculated by the ratio between urea and creatinine and quantified by the calorimetric biochemical method and CRP by the immunoturbidimetric method. Differences between groups were applied by the Mann-Whitney U test and the automated binary regression test. RESULTS: Of the 589 patients, 87.4% were at nutritional risk. When evaluating patients admitted to the ICU, 91.9% were at nutritional risk. Patients with NRS ≥3 are older, with lower body mass and BMI, higher NLR and UCR and lower CRP values. However, 73% of patients admitted to the ward were at nutritional risk, and only age differed between groups, being higher in patients with NRS ≥3. Logistic regression showed a weak association between nutritional risk in NRS and UCR (Model 1) (OR = 0.96, 95%CI: 0.94-0.99, p = 0.003) and NRS with CRP (Model 1) (OR = 1.01, 95%CI: 1.00-1.02, p < 0.001) in patients in the ICU. On the other hand, the logistic regression in ward patients found association only for CRP in both models (Model 1, OR = 1.01, 95%CI: 1.00-1.01, p = 0.041) and (Model 2, OR = 1.01, 95%CI: 1.00-1.01, p = 0.031). CONCLUSION: We found a weak association between nutritional risk and UCR and CRP levels in patients admitted to the ICU, while in the ward patients the nutritional risk also had weak association with CRP.
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Biomarcadores , Proteína C-Reactiva , COVID-19 , Creatinina , Hospitalización , Estado Nutricional , SARS-CoV-2 , Urea , Humanos , COVID-19/sangre , Masculino , Femenino , Estudios Transversales , Proteína C-Reactiva/metabolismo , Proteína C-Reactiva/análisis , Persona de Mediana Edad , Anciano , Estudios Retrospectivos , Creatinina/sangre , Urea/sangre , Biomarcadores/sangre , Evaluación Nutricional , Neutrófilos , Anciano de 80 o más Años , Factores de Riesgo , Adulto , Desnutrición/sangreRESUMEN
The outbreak of the disease caused by the SARS-CoV-2 virus (Covid-19) has resulted in a global health emergency that has caused millions of deaths in recent years. The control of the pandemic was significantly impacted by the availability of inputs and qualified labor to correctly diagnose the population. The challenges faced by numerous countries in conducting this extensive diagnosis, utilizing methods such as RT-PCR, emphasize the necessity for alternative testing strategies that are less reliant on expensive raw materials and can be implemented on a larger scale. This paper proposes a methodology for classifying blood serum samples as either positive or negative for Covid-19 infection using excitation-emission matrix (EEM) fluorescence spectroscopy associated with multivariate analysis. The proposed methodology uses EEM spectra of samples diagnosed by the reference method (RT-PCR) to train and validate classification models. Two approaches were tested: the first using PARAFAC and the second by unfolding the excitation-emission matrices. The DD-SIMCA model performed best in the PARAFAC approach, with an error rate of 0.05, sensitivity of 0.98 and specificity of 0.96. The PLS-DA and PCA-DA models in the second approach effectively distinguished between classes. The PCA-DA model performed the best with an error rate of 0.06 and sensitivity and specificity of 0.94. Fluorescence spectroscopy was found to be effective in analyzing serum samples and obtaining discrimination models to determine if a patient is infected with SARS-CoV-2. The findings are encouraging and could aid in the development of an inexpensive and reliable auxiliary diagnostic method.
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COVID-19 , SARS-CoV-2 , Espectrometría de Fluorescencia , Humanos , Espectrometría de Fluorescencia/métodos , COVID-19/diagnóstico , COVID-19/sangre , SARS-CoV-2/aislamiento & purificación , Análisis Discriminante , Análisis Multivariante , Análisis de los Mínimos Cuadrados , Análisis de Componente PrincipalRESUMEN
BACKGROUND: The prevalence of COVID-19 as the primary diagnosis among hospitalized patients with myocardial injury has increased during the pandemic and targeting elevated oxidant stress and inflammatory biomarkers may offer a potential role for novel therapies to improve outcomes. METHODS: At a single VA Medical Center from January 1 through December 31, 2021, troponin assays from patients being evaluated in the Emergency Room for consideration of admission were analyzed and peak levels from each patient were considered abnormal if exceeding the Upper Reference Limit (URL). Among admitted patients with an elevated troponin level, ICD-10 diagnoses were categorized, biomarker elevations were recorded, and independent predictors of death in patients with COVID-19 were determined at a median of 6-months following admission. RESULTS: Of 998 patients, 399 (40 %) had a negative troponin and were not included in the analysis. Additional patients with an elevated troponin were also excluded, either because they were not admitted (n = 68) or had a final diagnosis of Type 1 MI (n = 117). Of the remaining 414 patients with an elevated peak troponin, COVID-19 was the primary diagnosis in 43 patients (10 %) and was the 4th most common diagnosis of patients admitted with myocardial injury behind congestive heart failure, sepsis, and COPD or pneumonia. At a median of 6-months following admission, 18 (42 %) of the COVID-19 patients had died and independent predictors of death (Odd Ratio: Confidence Intervals) were age (1.18: 1.06â1.37), Troponin level (Log 10 transformed) (16.54: 2.30â266.65) and C-Reactive Protein (CRP) (1.30: 1.10â1.65). CONCLUSIONS: Newly diagnosed COVID-19 during the pandemic was a common cause of elevated troponin in hospitalized patients without a Type 1 MI. Age, peak troponin level and peak CRP level were independent predictors of poor outcomes and suggest a need to target these cardiac biomarkers, potentially with novel antioxidant or anti-inflammatory therapies.
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Biomarcadores , COVID-19 , Troponina , Humanos , COVID-19/sangre , COVID-19/mortalidad , Biomarcadores/sangre , Masculino , Femenino , Persona de Mediana Edad , Anciano , Troponina/sangre , SARS-CoV-2 , Infarto del Miocardio/sangre , Infarto del Miocardio/diagnósticoRESUMEN
Platelets are enucleated fragments of cells with a diversity of internal granules. They are responsible for functions related to hemostasis, coagulation, and inflammation. The activation of these processes depends on a cascade coordinated by cytokines, chemokines, and components of purinergic signaling, such as ATP, ADP, and adenosine. Platelets express distinct components of the purinergic system: P2X1, P2Y1, PY12, and P2Y14 receptors; and the ectonucleotidases NTPDase, NPP, and 5NTE (ecto-5'-nucleotidase). Except for P2Y14, which has not yet exhibited a known function, all other components relate to the biological processes mentioned before. Platelets are known to display specific responses to microorganisms, being capable of recognizing pathogen-associated molecular patterns (PAMPs), engulfing certain classes of viruses, and participating in NETosis. Platelet function dysregulation implicates various pathophysiological processes, including cardiovascular diseases (CVDs) and infections. In COVID-19 patients, platelets exhibit altered purinergic signaling and increased activation, contributing to inflammation. Excessive platelet activation can lead to complications from thrombosis, which can affect the circulation of vital organs. Therefore, controlling the activation is necessary to end the inflammatory process and restore homeostasis. Ectonucleotidases, capable of hydrolyzing ATP, ADP, and AMP, are of fundamental importance in activating platelets, promising pharmacological targets for clinical use as cardiovascular protective drugs. In this review, we revisit platelet biology, the purinergic receptors and ectonucleotidases on their surface, and their importance in platelet activity. Additionally, we describe methods for isolating platelets in humans and murine, as well as the main techniques for detecting the activity of ectonucleotidases in platelets. Considering the multitude of functions revealed by platelets and their potential use as potent bioreactors able to secrete and present molecules involved in the communication of the vasculature with the immune system, it is crucial to deeply understand platelet biology and purinergic signaling participation to contribute to the developing of therapeutic strategies in diseases of the cardiovascular, inflammatory, and immune systems.
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Plaquetas , COVID-19 , Activación Plaquetaria , Animales , Humanos , 5'-Nucleotidasa/metabolismo , Adenosina Trifosfatasas/metabolismo , Plaquetas/metabolismo , Plaquetas/inmunología , Separación Celular/métodos , COVID-19/inmunología , COVID-19/sangre , Activación Plaquetaria/inmunología , SARS-CoV-2/inmunología , Transducción de SeñalRESUMEN
INTRODUCTION: Despite its crucial role in Epidermal Growth Factor Receptor (EGFR) activation, and the resulting impact on the health-disease process, epidermal growth factor (EGF) is an underexplored molecule in relation to how its serum concentrations relate to other analytes and clinical variables in pathological contexts. OBJECTIVE: To clarify the possible correlation between EGF and clinical and analytical variables in the context of COVID-19. METHODS: Cross-sectional observational and analytical study, in patients with virological and clinical diagnosis of COVID-19, selected by simple random sampling, admitted between August and September 2021. UMELISA-EGF commercial kits were used. RESULTS: Differences in overall EGF values were observed between groups (566.04 vs. 910.53 pg/ml, p = .0430). In COVID-19 patients, no notable correlations were observed for neutrophil, platelet, triglyceride or liver enzyme values (p > .05). Significant correlations were observed with the neutrophil-lymphocyte indicator (r = 0.4711, p = .0128) as well as with the platelet-lymphocyte index (r = 0.4553, p = .0155). Statistical results of multivariate regression analysis suggest NLR (ß = .2232, p = .0353) and PLR (ß = .2117, p = .0411) are predictors of inflammation in patients with COVID-19. CONCLUSIONS: Serum EGF concentrations in COVID-19 correlate positively with prognostic inflammatory markers of severity and could presumably act as an independent risk factor for the development of inflammation in response to new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).
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COVID-19 , Factor de Crecimiento Epidérmico , Inflamación , SARS-CoV-2 , Humanos , COVID-19/sangre , COVID-19/diagnóstico , Factor de Crecimiento Epidérmico/sangre , Masculino , Femenino , Estudios Transversales , Persona de Mediana Edad , Anciano , Inflamación/sangre , Adulto , Biomarcadores/sangre , Neutrófilos/inmunologíaRESUMEN
The IgG response against SARS-CoV-2 infection can persist for over six months (long response; LR). However, among 30% of those infected, the duration can be as short as three months or less (short response; SR). The present study assembled serological data on the anti-SARS-CoV-2 IgG response duration of two previous studies and integrated these results with the plasmatic cytokine levels and genetic profile of 10 immune-relevant SNPs that were also previously published, along with the plasmatic total IgG, IgA, and IgM levels, allowing for the genetic, clinical, immunological, and epidemiological aspects of the post-COVID-19 IgG response duration to be understood. The SR was associated with previous mild acute COVID-19 and with an SNP (rs2228145) in IL6R related to low gene expression. Additionally, among the SR subgroup, no statistically significant Spearman correlations were observed between the plasma levels of IL-17A and the Th17 regulatory cytokines IFN-γ (rs = 0.2399; p = 0.1043), IL-4 (rs = 0.0273; p = 0.8554), and IL-2 (rs = 0.2204; p = 0.1365), while among the LR subgroup, weaker but statistically significant Spearman correlations were observed between the plasma levels of IL-17A and IFN-γ (rs = 0.3873; p = 0.0016), IL-4 (rs = 0.2671; p = 0.0328), and IL-2 (rs = 0.3959; p = 0.0012). These results suggest that the Th17 response mediated by the IL-6 pathway has a role in the prolonged IgG response to SARS-CoV-2 infection.
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Anticuerpos Antivirales , COVID-19 , Inmunoglobulina G , Polimorfismo de Nucleótido Simple , SARS-CoV-2 , Humanos , COVID-19/inmunología , COVID-19/epidemiología , COVID-19/sangre , COVID-19/virología , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , SARS-CoV-2/inmunología , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Masculino , Femenino , Receptores de Interleucina-6/genética , Persona de Mediana Edad , Adulto , Interleucina-17/sangre , Interleucina-17/genética , Citocinas/sangre , Inmunoglobulina A/sangre , Interferón gamma/sangre , Interferón gamma/genética , Inmunoglobulina M/sangre , Interleucina-4/sangre , Interleucina-4/genética , AncianoRESUMEN
OBJECTIVE: To investigate the relationship between the levels of adipokines and other endocrine biomarkers and patient outcomes in hospitalized patients with COVID-19. METHODS: In a prospective study that included 213 subjects with COVID-19 admitted to the intensive care unit, we measured the levels of cortisol, C-peptide, glucagon-like peptide-1, insulin, peptide YY, ghrelin, leptin, and resistin.; their contributions to patient clustering, disease severity, and predicting in-hospital mortality were analyzed. RESULTS: Cortisol, resistin, leptin, insulin, and ghrelin levels significantly differed between severity groups, as defined by the World Health Organization severity scale. Additionally, lower ghrelin and higher cortisol levels were associated with mortality. Adding biomarkers to the clinical predictors of mortality significantly improved accuracy in determining prognosis. Phenotyping of subjects based on plasma biomarker levels yielded two different phenotypes that were associated with disease severity, but not mortality. CONCLUSION: As a single biomarker, only cortisol was independently associated with mortality; however, metabolic biomarkers could improve mortality prediction when added to clinical parameters. Metabolic biomarker phenotypes were differentially distributed according to COVID-19 severity but were not associated with mortality.
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Biomarcadores , COVID-19 , Fenotipo , Humanos , COVID-19/sangre , COVID-19/mortalidad , COVID-19/diagnóstico , Biomarcadores/sangre , Masculino , Femenino , Estudios Prospectivos , Pronóstico , Persona de Mediana Edad , Anciano , Índice de Severidad de la Enfermedad , Unidades de Cuidados Intensivos , Hidrocortisona/sangre , Mortalidad Hospitalaria , SARS-CoV-2RESUMEN
BACKGROUND: SARS-CoV-2 is a systemic disease that affects endothelial function and leads to coagulation disorders, increasing the risk of mortality. Blood levels of endothelial biomarkers such as Von Willebrand Factor (VWF), Thrombomodulin or Blood Dendritic Cell Antigen-3 (BDCA3), and uUokinase (uPA) increase in patients with severe disease and can be prognostic indicators for mortality. Therefore, the aim of this study was to determine the effect of VWF, BDCA3, and uPA levels on mortality. METHODS: From May 2020 to January 2021, we studied a prospective cohort of hospitalized adult patients with polymerase chain reaction (PCR)-confirmed COVID-19 with a SaO2 ≤ 93% and a PaO2/FiO2 ratio < 300. In-hospital survival was evaluated from admission to death or to a maximum of 60 days of follow-up with Kaplan-Meier survival curves and Cox proportional hazard models as independent predictor measures of endothelial dysfunction. RESULTS: We recruited a total of 165 subjects (73% men) with a median age of 57.3 ± 12.9 years. The most common comorbidities were obesity (39.7%), hypertension (35.4%) and diabetes (30.3%). Endothelial biomarkers were increased in non-survivors compared to survivors. According to the multivariate Cox proportional hazard model, those with an elevated VWF concentration ≥ 4870 pg/ml had a hazard ratio (HR) of 4.06 (95% CI: 1.32-12.5) compared to those with a lower VWF concentration adjusted for age, cerebrovascular events, enoxaparin dose, lactate dehydrogenase (LDH) level, and bilirubin level. uPA and BDCA3 also increased mortality in patients with levels ≥ 460 pg/ml and ≥ 3600 pg/ml, respectively. CONCLUSION: The risk of mortality in those with elevated levels of endothelial biomarkers was observable in this study.