Leukocyte surface expression of the endoplasmic reticulum chaperone GRP78 is increased in severe COVID-19.

Hyperinflammation present in individuals with severe COVID-19 has been associated with an exacerbated cytokine production and hyperactivated immune cells. Endoplasmic reticulum stress leading to the unfolded protein response has been recently reported as an active player in inducing inflammatory responses. Once unfolded protein response is activated, GRP78, an endoplasmic reticulum-resident chaperone, is translocated to the cell surface (sGRP78), where it is considered a cell stress marker; however, its presence has not been evaluated in immune cells during disease. Here we assessed the presence of sGRP78 on different cell subsets in blood samples from severe or convalescent COVID-19 patients. The frequency of CD45+sGRP78+ cells was higher in patients with the disease compared to convalescent patients. The latter showed similar frequencies to healthy controls. In patients with COVID-19, the lymphoid compartment showed the highest presence of sGRP78+ cells versus the myeloid compartment. CCL2, TNF-α, C-reactive protein, and international normalized ratio measurements showed a positive correlation with the frequency of CD45+sGRP78+ cells. Finally, gene expression microarray data showed that activated T and B cells increased the expression of GRP78, and peripheral blood mononuclear cells from healthy donors acquired sGRP78 upon activation with ionomycin and PMA. Thus, our data highlight the association of sGRP78 on immune cells in patients with severe COVID-19.

Strategies to recover blood donors during the COVID-19 pandemic: a tertiary level hospital experience.

BACKGROUND: The SARS-CoV-2 pandemic has challenged blood banks. In Mexico, donors decreased 22% between April and May 2020 compared to the same months in 2019. This study analyzed the effect of the strategies to recover donors (altruistic and family) in a tertiary pediatric care center during the pandemic. METHODS: The Blood Bank of the Hospital Infantil de México Federico Gómez implemented strategies to obtain blood components to ensure self-sufficiency. The effect of these strategies on donor recovery was analyzed. RESULTS: There were 7,315 eligible donors in 2019 and 5,070 in 2020. Blood component requirements decreased from 10,037 units in 2019 to 8,619 in 2020. The strategies aimed at attracting altruistic donors managed to increase the percentage of this type of donor when comparing the months in which these strategies were applied with the same months in 2019. In addition, it was observed that the greater the number of methods used simultaneously, the higher the percentage of altruistic donors (rho = 0.846, p = 0.002). In contrast, strategies aimed at attracting family donors did not increase the number of this type of donor. CONCLUSIONS: Actions to recruit altruistic donors increased the number of this type of donor to meet the hospital's needs.

INTRODUCCIÓN: La pandemia por SARS-CoV-2 ha representado un reto en los bancos de sangre. En México, los donadores disminuyeron el 22% entre abril y mayo del 2020 en comparación con los mismos meses del 2019. Este estudio analizó el efecto de las estrategias realizadas para recuperar donantes, altruistas y familiares, en un centro de atención pediátrica de tercer nivel durante la pandemia. MÉTODOS: El Banco de Sangre del Hospital Infantil de México Federico Gómez implementó estrategias encaminadas a la obtención de componentes sanguíneos para asegurar la autosuficiencia. Se analizó el efecto de dichas estrategias en la recuperación de donantes. RESULTADOS: Se registraron 7,315 donadores aptos en el año 2019 y 5,070 en el 2020. Los requerimientos de componentes sanguíneos disminuyeron de 10,037 unidades en 2019 a 8,619 en 2020. Las estrategias que estaban destinadas a captar donadores altruistas lograron aumentar el porcentaje de este tipo de donadores al comparar los meses en que se aplicaron dichas estrategias con los mismos meses en el 2019. Además, se observó que, a mayor número de estrategias aplicadas de manera simultánea, mayor porcentaje de donadores altruistas (rho = 0.846, p = 0.002). Por el contrario, las estrategias con la finalidad de atraer donadores familiares no lograron aumentar la cantidad de este tipo de donadores. CONCLUSIONES: Las acciones para recabar donadores altruistas aumentaron la cantidad de este tipo de donadores para satisfacer las necesidades del hospital.

Efficacy and safety of vitamin D supplementation in hospitalized COVID-19 pediatric patients: A randomized controlled trial.

Background: Some studies suggested that adequate levels of vitamin D (VD) decrease the risk of severe COVID-19. Information about the effectiveness of VD supplementation in children is scarce. Objective: To assess the efficacy and safety of VD supplementation compared to the standard of care in hospitalized children with COVID-19. Patients and methods: An open-label randomized controlled single-blind clinical trial was carried out. We included patients from 1 month to 17 years, with moderate COVID-19, who required hospitalization and supplemental oxygen. They were randomized into two groups: the VD group, which received doses of 1,000 (children < 1 year) or 2,000 IU/day (from 1 to 17 years) and the group without VD (control). The outcome variables were the progression of oxygen requirement, the development of complications, and death. Statistical analysis: For comparison between groups, we used the chi-squared test or Fisher's exact test and the Mann-Whitney U test. Absolute risk reduction (ARR) and the number needed to treat (NNT) were calculated. p ≤ 0.05 was considered statistically significant. Results: From 24 March 2020 to 31 March 2021, 87 patients were eligible to participate in the trial; 45 patients were randomized: 20 to the VD group and 25 to the control group. There was no difference in general characteristics at baseline, including serum VD levels (median 13.8 ng/ml in the VD group and 11.4 ng/ml in the control group). Outcomes: 2/20 (10%) in the VD group vs. 9/25 (36%) in the control group progressed to a superior ventilation modality (p = 0.10); one patient in the VD group died (5%) compared to 6 (24%) patients in the control group (p = 0.23). ARR was 26% (95% CI 8.8 to 60.2%) and NNT was 3 (2 to 11) for progression and ARR was 19% (95% CI -3.9 to 42.8%) and NNT was 6 (2 to 26) for death. None of the patients receiving VD had adverse effects. The trial was stopped for ethical reasons; since after receiving the results of the basal VD values, none of the patients had normal levels. Conclusion: In this trial, VD supplementation in pediatric patients seems to decrease the risk of COVID-19 progression and death. More studies are needed to confirm these findings. Clinical Trial Registration: This protocol was registered on ClinicalTrials.gov with the registration number NCT04502667.

25-Hydroxyvitamin D level is associated with mortality in patients with critical COVID-19: a prospective observational study in Mexico City.

BACKGROUND/OBJECTIVES: Considering the high number of deaths from coronavirus disease 2019 (COVID-19) in Latin American countries, together with multiple factors that increase the prevalence of vitamin D deficiency, we aimed to determine 25-hydroxyvitamin D (25[OH]D) levels and its association with mortality in patients with critical COVID-19. SUBJECTS/METHODS: This was a prospective observational study including adult patients with critical COVID-19. Data, including clinical characteristics and 25(OH)D levels measured at the time of intensive care unit admission, were collected. All patients were followed until hospital discharge or in-hospital death. The patients were divided into those surviving and deceased patient groups, and univariate and multivariate logistic regression analyses were performed to determine independent predictors of in hospital mortality. RESULTS: The entire cohort comprised 94 patients with critical COVID-19 (males, 59.6%; median age, 61.5 years). The median 25(OH)D level was 12.7 ng/mL, and 15 (16%) and 79 (84%) patients had vitamin D insufficiency and vitamin D deficiency, respectively. The median serum 25(OH)D level was significantly lower in deceased patients compared with surviving (12.1 vs. 18.7 ng/mL, P < 0.001). Vitamin D deficiency was present in 100% of the deceased patients. Multivariate logistic regression analysis revealed that age, body mass index, other risk factors, and 25(OH)D level were independent predictors of mortality. CONCLUSIONS: Vitamin D deficiency was present in 84% of critical COVID-19 patients. Serum 25(OH)D was independently associated with mortality in critical patients with COVID-19.

Analysis of the Behaviour of Immunoglobulin G Antibodies in Children and Adults Convalescing From Severe Acute Respiratory Syndrome-Coronavirus-2 Infection.

The pandemic caused by SARS CoV-2 (COVID-19) has affected millions of people since 2020. There are clinical differences and in mortality between the adult and paediatric population. Recently, the immune response through the development of antibodies has gained relevance due to the risk of reinfection and vaccines' development. Objective: Was to compare the association of clinical history and the clinical presentation of the disease with the development of IgG antibodies against SARS-CoV-2 in paediatric and adult patients with a history of positive reverse transcriptase-polymerase chain reaction (RT-PCR) results. Methods: Cross-sectional observational study carried out in a Paediatric Hospital in Mexico City included patients under 18 years of age and health personnel with positive RT-PCR for COVID-19 comparing antibody expression. The development of specific IgG antibodies was measured, the presence of comorbidities, duration, and severity of symptoms was determined. Results: Sixty-one subjects (20 < 18 years and 41 > 18 years) were analysed. The median sample collection was 3 weeks. There were no differences in the expression of specific antibodies; no differences were shown according to the symptoms' severity. A positive correlation (r = 0.77) was demonstrated between the duration of symptoms and antibody levels. Conclusions: In conclusion, there is a clear association between the duration of the symptoms associated with SARS-CoV-2 infection and the IgG units generated in paediatric and adult patients convalescing from COVID-19.

Risk conditions in healthcare workers of a pediatric coronavirus disease center in Mexico City.

BACKGROUND: The new evere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is characterized by its high capacity to transmit. Health-care personnel is highly susceptible to becoming infected. This study aimed to determine the characteristics and known risk factors for contagion and severe outcomes of SARS-CoV-2 disease in health-care personnel of a pediatric coronavirus disease (COVID) center in Mexico City. METHODS: In the last week of March 2020 (at the beginning of phase 2 of the Ministry of Health's national campaign in Mexico), a study was conducted on healthcare workers of a pediatric COVID hospital in Mexico City. Using a virtual interview, we evaluated comorbidities, mobility, areas and functions where they carry out the activities, protection measures, contact history, and vaccination. According to their activities, healthcare workers were classified into the following areas: medical, nursing, other health-care personnel (researchers, nutritionists, rehabilitation, imaging, and laboratory), administrative, and other services. We compared the variables between the groups of healthcare workers with the X2 test. RESULTS: We included 812 participants. The mean age was 41 ± 11 years, and 33% were overweight or obese, 18% were over 60 years old, and 19% had high blood pressure. Medical and nursing personnel presented a higher proportion in the use of standard protection measures. CONCLUSIONS: Among healthcare workers, there are risk conditions for the development of complications in case of SARS-CoV-2 infection. Most medical and nursing personnel use standard protective measures.

Analytical recommendations for SARS-CoV-2 identification by RT-PCR in pediatric patients.

Coronavirus disease 2019 (COVID-19) is caused by the severe acute respiratory syndrome 2 coronavirus (SARS-CoV-2) and is currently listed as a global public health emergency. Timely identification and protocol implementations for molecular detection of this virus are vital for medical decision-making. Identification of SARS-CoV-2 infection cases is based on detection of the virus RNA by molecular tests, particularly real-time reverse transcription-polymerase chain reaction (RT-PCR). Technical and operational details specific to each center must be considered to perform the molecular diagnosis of SARS-CoV-2 in pediatric patients. The term "qualified laboratories" involves laboratories in which all users, analysts, and anyone reporting results are trained to develop and interpret results through a procedure implemented previously by an instructor. Such knowledge is essential in detecting and identifying errors during each of its phases: pre-analytical, analytical, and post-analytical, which allow the establishment of continuous improvement policies to ensure the quality of the results, but above all, the physical integrity of health workers.

Genotyping of the Major SARS-CoV-2 Clade by Short-Amplicon High-Resolution Melting (SA-HRM) Analysis.

The genome of the SARS-CoV-2 virus, the causal agent of the COVID-19 pandemic, has diverged due to multiple mutations since its emergence as a human pathogen in December 2019. Some mutations have defined several SARS-CoV-2 clades that seem to behave differently in terms of regional distribution and other biological features. Next-generation sequencing (NGS) approaches are used to classify the sequence variants in viruses from individual human patients. However, the cost and relative scarcity of NGS equipment and expertise in developing countries prevent studies aimed to associate specific clades and variants to clinical features and outcomes in such territories. As of March 2021, the GR clade and its derivatives, including the B.1.1.7 and B.1.1.28 variants, predominate worldwide. We implemented the post-PCR small-amplicon high-resolution melting analysis to genotype SARS-CoV-2 viruses isolated from the saliva of individual patients. This procedure was able to clearly distinguish two groups of samples of SARS-CoV-2-positive samples predicted, according to their melting profiles, to contain GR and non-GR viruses. This grouping of the samples was validated by means of amplification-refractory mutation system (ARMS) assay as well as Sanger sequencing.

Restructuring of a pediatric hospital in the face of the COVID-19 pandemic.

Overview of the pandemic In December 2019, a new virus named SARS-CoV-2 was reported in Wuhan province, China. The first case of COVID-19 in Mexico was confirmed on February 28, 2020, and the World Health Organization declared the pandemic on March 11.

Saliva as a promising biofluid for SARS-CoV-2 detection during the early stages of infection.

Background: Diagnostic testing for coronavirus disease (COVID)-19 is performed using nasopharyngeal swabs. This type of sampling is uncomfortable for the patient, dangerous for health workers, and its high demand has led to a global shortage of swabs. One of the alternative specimens is saliva. However, the optimal conditions for the test have not been established. Methods: Reverse transcription-polymerase chain reaction was used to detect the viral genome in saliva samples kept at room temperature, in the fridge or frozen for 2 days. In addition, the influence of brushing teeth and feeding on the detection of the virus in saliva was addressed. Finally, the efficiency of saliva in revealing the presence of the virus during the hospitalization period was determined in children. Results: The viral genome was consistently detected regardless of the storage conditions of saliva samples. Brushing teeth and feeding did not influence the sensitivity of the test. In hospitalized children, positive results were obtained only during the early days. Conclusions: These results support the idea of the use of saliva as an alternative specimen for diagnostic testing for COVID-19. The viral genome is stable and endures perturbations in the oral cavity. However, clearance of the virus from the mouth during the infection may limit the use of the test only to the early stages of the disease.

Introducción: El diagnóstico de COVID-19 (enfermedad por coronavirus 2019) se realiza con un hisopado nasofaríngeo. El procedimiento de toma de muestra es molesto para el paciente y peligroso para el personal de salud, y la alta demanda de análisis ha conducido a la escasez de hisopos. Una alternativa es el uso de saliva, pero las condiciones óptimas para realizar el estudio no han sido establecidas. Métodos: Se usó la reacción en cadena de la polimerasa con transcriptasa reversa para detectar el genoma viral en muestras de saliva mantenidas a temperatura ambiente, en refrigeración o congeladas. Además, se evaluó la influencia del aseo bucal y de la ingesta de alimento en la detección del virus. Finalmente, se determinó el desempeño de la saliva para reportar la presencia del virus durante el periodo de hospitalización en niños. Resultados: El genoma viral fue estable durante 2 días a las diferentes temperaturas ensayadas. El aseo bucal y la ingesta de alimento no influyeron en la detección del virus. En los niños hospitalizados solo se obtuvieron resultados positivos durante los primeros días. Conclusiones: Los resultados coinciden con la idea del uso de la saliva como biofluido alternativo para el diagnóstico de COVID-19. El genoma viral es estable y no se ve afectado por perturbaciones en la cavidad oral; sin embargo, la dinámica de la infección puede provocar que el ensayo solo sea útil durante las primeras etapas de la enfermedad.

Clinical risk profile associated with SARS-CoV-2 infection and complications in the emergency area of a pediatric COVID-19 center.

Background: In February 2020, the disease caused by the novel coronavirus (SARS-CoV-2), was classified as a pandemic. In the pediatric population, coronavirus disease (COVID)-19 has a reported mortality of less than 6% in complicated cases; however, the clinical characteristics and severity are not the same as those presented in the adult population. This study aimed to describe the clinical manifestations of patients younger than 18 years old and their association with the confirmation of the test and outcomes. Methods: We conducted an analytical cross-sectional study of symptoms suggestive for SARS-CoV-2 infection. All subjects with a confirmatory test for SARS-CoV-2 were included. Initial symptoms, history of influenza vaccination, and previous contact were documented, and mortality and the requirement for assisted mechanical ventilation were identified. The proportions of the variables were compared with the χ2 test. The odds ratio for a positive test and the requirement of intubation was calculated. Results: Of a total of 510 subjects, 76 (15%) were positive for SARS-CoV-2. The associated symptoms were chest pain, sudden onset of symptoms, and general malaise. The variable most associated with contagion was the exposure to a relative with a confirmed diagnosis of COVID-19. Infants and subjects without the influenza vaccine showed an increased risk for respiratory complications. Conclusions: The frequency of positivity in the test was 15% (infants and adolescents represented 64% of the confirmed cases), and the associated factors identified were contact with a confirmed case, sudden onset of symptoms, and chest pain.

Introducción: En 2019 se reportaron los primeros casos de SARS-CoV-2 (coronavirus tipo 2 del síndrome respiratorio agudo grave), causante de la COVID-19, que alcanzó el grado de pandemia en febrero de 2020. La presentación en la etapa pediátrica reporta una mortalidad menor del 6% en los casos complicados; sin embargo, las características clínicas y su gravedad no son iguales que en la población adulta. El objetivo de este estudio fue describir las manifestaciones clínicas de los pacientes menores de 18 años y su asociación con la confirmación de la prueba, la intubación endotraqueal y la muerte. Métodos: Estudio transversal analítico por cuadro sugestivo de infección por SARS-CoV-2. Se incluyeron sujetos positivos para SARS-CoV-2. Se documentaron los síntomas iniciales, los antecedentes de vacunación contra la influenza y los contactos previos, y se identificaron los desenlaces de mortalidad y requerimiento de ventilación mecánica asistida. Se compararon las proporciones de las variables con la prueba χ2 y se calculó la razón de momios para la presencia de una prueba positiva y requerir intubación. Resultados: De un total de 510 sujetos, 76 (15%) fueron positivos para SARS-CoV-2. Los síntomas asociados fueron dolor precordial, inicio súbito y malestar general. La variable asociada con mayor frecuencia el contagio fue la exposición a un familiar con COVID-19 confirmada. Los sujetos sin vacuna de la influenza presentaron un riesgo mayor de complicaciones respiratorias. Conclusiones: La frecuencia de positividad en la prueba fue del 15%. Se identificaron como factores asociados a prueba positiva el contacto con un caso confirmado de COVID-19, el inicio súbito de los síntomas y el dolor precordial.

Clinical risk profile associated with SARS-CoV-2 infection and complications in the emergency area of a pediatric COVID-19 center

Background: In February 2020, the disease caused by the novel coronavirus (SARS-CoV-2), was classified as a pandemic. In the pediatric population, coronavirus disease (COVID)-19 has a reported mortality of less than 6% in complicated cases;however, the clinical characteristics and severity are not the same as those presented in the adult population. This study aimed to describe the clinical manifestations of patients younger than 18 years old and their association with the confirmation of the test and outcomes. Methods: We conducted an analytical cross-sectional study of symptoms suggestive for SARS-CoV-2 infection. All subjects with a confirmatory test for SARS-CoV-2 were included. Initial symptoms, history of influenza vaccination, and previous contact were documented, and mortality and the requirement for assisted mechanical ventilation were identified. The proportions of the variables were compared with the &#967;2 test. The odds ratio for a positive test and the requirement of intubation was calculated. Results: Of a total of 510 subjects, 76 (15%) were positive for SARS-CoV-2. The associated symptoms were chest pain, sudden onset of symptoms, and general malaise. The variable most associated with contagion was the exposure to a relative with a confirmed diagnosis of COVID-19. Infants and subjects without the influenza vaccine showed an increased risk for respiratory complications. Conclusions: The frequency of positivity in the test was 15% (infants and adolescents represented 64% of the confirmed cases), and the associated factors identified were contact with a confirmed case, sudden onset of symptoms, and chest pain. Resumen Introducción: En 2019 se reportaron los primeros casos de SARS-CoV-2 (coronavirus tipo 2 del síndrome respiratorio agudo grave), causante de la COVID-19, que alcanzó el grado de pandemia en febrero de 2020. La presentación en la etapa pediátrica reporta una mortalidad menor del 6% en los casos complicados;sin embargo, las características clínicas y su gravedad no son iguales que en la población adulta. El objetivo de este estudio fue describir las manifestaciones clínicas de los pacientes menores de 18 años y su asociación con la confirmación de la prueba, la intubación endotraqueal y la muerte. Métodos: Estudio transversal analítico por cuadro sugestivo de infección por SARS-CoV-2. Se incluyeron sujetos positivos para SARS-CoV-2. Se documentaron los síntomas iniciales, los antecedentes de vacunación contra la influenza y los contactos previos, y se identificaron los desenlaces de mortalidad y requerimiento de ventilación mecánica asistida. Se compararon las proporciones de las variables con la prueba &#967;2 y se calculó la razón de momios para la presencia de una prueba positiva y requerir intubación. Resultados: De un total de 510 sujetos, 76 (15%) fueron positivos para SARS-CoV-2. Los síntomas asociados fueron dolor precordial, inicio súbito y malestar general. La variable asociada con mayor frecuencia el contagio fue la exposición a un familiar con COVID-19 confirmada. Los sujetos sin vacuna de la influenza presentaron un riesgo mayor de complicaciones respiratorias. Conclusiones: La frecuencia de positividad en la prueba fue del 15%. Se identificaron como factores asociados a prueba positiva el contacto con un caso confirmado de COVID-19, el inicio súbito de los síntomas y el dolor precordial.

Saliva as a promising biofluid for SARS-CoV-2 detection during the early stages of infection

Background: Diagnostic testing for coronavirus disease (COVID)-19 is performed using nasopharyngeal swabs. This type of sampling is uncomfortable for the patient, dangerous for health workers, and its high demand has led to a global shortage of swabs. One of the alternative specimens is saliva. However, the optimal conditions for the test have not been established. Methods: Reverse transcription-polymerase chain reaction was used to detect the viral genome in saliva samples kept at room temperature, in the fridge or frozen for 2 days. In addition, the influence of brushing teeth and feeding on the detection of the virus in saliva was addressed. Finally, the efficiency of saliva in revealing the presence of the virus during the hospitalization period was determined in children. Results: The viral genome was consistently detected regardless of the storage conditions of saliva samples. Brushing teeth and feeding did not influence the sensitivity of the test. In hospitalized children, positive results were obtained only during the early days. Conclusions: These results support the idea of the use of saliva as an alternative specimen for diagnostic testing for COVID-19. The viral genome is stable and endures perturbations in the oral cavity. However, clearance of the virus from the mouth during the infection may limit the use of the test only to the early stages of the disease. Resumen Introducción: El diagnóstico de COVID-19 (enfermedad por coronavirus 2019) se realiza con un hisopado nasofaríngeo. El procedimiento de toma de muestra es molesto para el paciente y peligroso para el personal de salud, y la alta demanda de análisis ha conducido a la escasez de hisopos. Una alternativa es el uso de saliva, pero las condiciones óptimas para realizar el estudio no han sido establecidas. Métodos: Se usó la reacción en cadena de la polimerasa con transcriptasa reversa para detectar el genoma viral en muestras de saliva mantenidas a temperatura ambiente, en refrigeración o congeladas. Además, se evaluó la influencia del aseo bucal y de la ingesta de alimento en la detección del virus. Finalmente, se determinó el desempeño de la saliva para reportar la presencia del virus durante el periodo de hospitalización en niños. Resultados: El genoma viral fue estable durante 2 días a las diferentes temperaturas ensayadas. El aseo bucal y la ingesta de alimento no influyeron en la detección del virus. En los niños hospitalizados solo se obtuvieron resultados positivos durante los primeros días. Conclusiones: Los resultados coinciden con la idea del uso de la saliva como biofluido alternativo para el diagnóstico de COVID-19. El genoma viral es estable y no se ve afectado por perturbaciones en la cavidad oral;sin embargo, la dinámica de la infección puede provocar que el ensayo solo sea útil durante las primeras etapas de la enfermedad.