Multicenter clinical evaluation of a novel transcription-mediated amplification assay for SARS-CoV-2 molecular testing / Evaluación clínica multicéntrica de un nuevo ensayo de amplificación mediada por transcripción para la detección molecular de SARS-CoV-2

Introduction The onset and spread of COVID-19 pandemic has forced clinical laboratories to rapidly expand testing capacity for SARS-CoV-2. This study evaluates the clinical performance of the TMA Procleix SARS-CoV-2 assay in comparison to the RT-PCR assay Allplex™ SARS-CoV-2 for the qualitative detection of SARS-CoV-2 RNA. Methods Between November 2020 and February 2021, 610 upper-respiratory specimens received for routine SARS-CoV-2 molecular testing were prospectively collected and selected at the Hospital Universitari Vall d’Hebron and the Hospital Universitari Bellvitge in Barcelona, Spain. All samples were processed in parallel with the TMA and the RT-PCR assays, and results were compared. Discrepancies were retested by an additional RT-PCR method and the clinical history of these patients was reviewed. Results Overall, the level of concordance between both assays was 92.0% (κ, 0.772). Most discordant results (36/38, 94.7%) corresponded to samples testing positive with the TMA assay and negative with the RT-PCR method. Of these discrepant cases, most (28/36, 77.8%) were finally classified as confirmed or probable SARS-CoV-2 cases according to the discrepant analysis. Conclusion In conclusion, the TMA Procleix SARS-CoV-2 assay performed well for the qualitative detection of SARS-CoV-2 RNA in a multisite clinical setting. This novel TMA assay demonstrated a greater sensitivity in comparison to RT-PCR methods for the molecular detection of SARS-CoV-2. This higher sensitivity but also the qualitative feature of this detection of SARS-CoV-2 should be considered when making testing algorithm decisions (AU)

Introducción El inicio y la expansión de la pandemia por COVID-19 han forzado a los laboratorios clínicos a ampliar rápidamente la capacidad de detección de SARS-CoV-2. Evaluamos el rendimiento clínico del ensayo de TMA Procleix SARS-CoV-2 en comparación con el ensayo de RT-PCR Allplex™ SARS-CoV-2 para la detección cualitativa de ARN de SARS-CoV-2. Métodos Entre noviembre de 2020 y febrero de 2021 se seleccionaron prospectivamente 610 muestras del tracto respiratorio superior recibidas de rutina en el Hospital Universitario Vall d’Hebron y el Hospital Universitario de Bellvitge en Barcelona, España, para el diagnóstico molecular de SARS-CoV-2. Todas las muestras fueron procesadas en paralelo con los ensayos de TMA y RT-PCR, y se compararon los resultados. Las discrepancias se estudiaron por un método adicional de RT-PCR y se revisaron las historias clínicas de los pacientes. Resultados En general, la concordancia entre ambos ensayos fue del 92,0% (κ, 0,772). La mayoría de los casos discrepantes (36/38, 94,7%) correspondían a muestras positivas con el ensayo de TMA y negativas con el método de RT-PCR. De estos, la mayoría (28/36, 77,8%) fueron finalmente clasificados como casos confirmados o probables de SARS-CoV-2 de acuerdo al análisis de discrepantes. Conclusión El ensayo de TMA Procleix SARS-CoV-2 funcionó bien para la detección cualitativa de ARN de SARS-CoV-2 en un entorno clínico multicéntrico. Este ensayo TMA demostró una mayor sensibilidad en comparación con métodos de RT-PCR para la detección molecular de SARS-CoV-2. Esta mayor sensibilidad, pero también el carácter cualitativo de esta detección de SARS-CoV-2, se deben considerar en el diagnóstico de la infección (AU)

The importance of prevalence and pre-test probability on the microbiological diagnosis of SARS-CoV-2: the case of Spain in 2020 / La importancia de la prevalencia y de la probabilidad pre-test en el diagnóstico microbiológico de SARS-CoV-2: el caso de España en 2020

Objectives. The aim of this work was to estimate the con ditioned probability for the diagnosis of SARS-CoV-2 infection with reverse transcription polymerase chain reaction (RT-PCR), viral antigen rapid diagnostic tests (Ag-RDT), and antibody detection tests depending on the prevalence in the specific healthcare settings in Spain in 2020, and on the pre-test prob ability (PTP) according to the clinical situation, age and un known or close contacts of the patient. Material and methods. Performance parameters of tests were obtained from literature. Prevalence data and PTP were obtained from Spanish sources and a survey, respectively. The post-test probability is the positive predictive value (PPV) when test is positive. For negative result, we also calculated the probability of having the infection (false negatives). Results. For both RT-PCR and viral Ag-RDT, the lowest PPV values were for the population screenings. This strategy proved to be useful in ruling out infection but generates a high number of false positives. At individual level, both tools provided high PPV (≥ 97%) when the PTP values are over 35%. In seroprevalence studies, though the specificity of IgG alone tests is high, under low seroprevalence, false positives cannot be avoided. Total antibodies tests are useful for diagnosis of COVID-19 in those doubtful cases with RT-PCR or Ag-RDT tests being repeatedly negative. Conclusions. The interpretating of results depends not only on the accuracy of the test, but also on the prevalence of the infection in different settings, and the PTP associated to the patient before performing the test (AU)

bjetivos. En este trabajo estimamos la probabilidad con dicionada del diagnóstico de infección por SARS-CoV-2 con RT PCR, pruebas de antígenos virales (Ag-RDT) y pruebas de detec ción de anticuerpos, en función de la prevalencia en España en diferentes ámbitos durante 2020, y de la probabilidad pre-test (PPT) según la situación clínica, edad y contactos del paciente. Material y métodos. Los parámetros de rendimiento de las pruebas se obtuvieron de bibliografía. Los datos de preva lencia y PPT se obtuvieron de fuentes españolas y de una en cuesta, respectivamente. La probabilidad post-test es el valor predictivo positivo (VPP) cuando la prueba es positiva. Para el resultado negativo, también calculamos la probabilidad de te ner la infección (falsos negativos). Resultados. Tanto con RT-PCR como con Ag-RDT, los va lores más bajos de VPP se detectaron en los cribados poblacio nales, que demostraron ser útiles para descartar la infección, pero generan muchos falsos positivos. A nivel individual, am bas pruebas proporcionaron un VPP ≥ 97% cuando los valores de PPT son superiores al 35%. En estudios de seroprevalencia, aunque la especificidad de las pruebas de IgG sola es alta, si la seroprevalencia es baja, no se pueden evitar falsos positivos. Además, las pruebas de anticuerpos totales pueden ayudar al diagnóstico de COVID-19 en aquellos casos dudosos con prue bas de RT-PCR o Ag-RDT repetidamente negativas. Conclusiones. La interpretación de los resultados depen de no sólo del rendimiento de las pruebas, sino también de la prevalencia de la infección en diferentes ámbitos, y de la PPT asociada al paciente antes de realizar la prueba (AU)

Role of Hologic® Panther Aptima™ SARS-CoV-2 assay in the detection of SARS-CoV-2: screening or diagnostic technique? / Papel del ensayo Hologic® Panther Aptima™ SARS-CoV-2 en la detección del SARS-CoV-2: ¿técnica de cribado o de diagnóstico?

During the multiple waves of COVID-19 suffered all over the world, having a rapid and sensitive diagnostic test has be come a priority for microbiology laboratories. The AptimaTM SARS-CoV-2 transcription-mediated amplification (TMA) assay running on the Panther system (Hologic) was presented as a very good option to cover this need. To evaluate this system, 570 respiratory samples were included in the study and were processed both by the Panther (Hologic) system and by qRT PCR (Thermo Fisher Science, Waltham, USA), current assay for the diagnosis of severe acute respiratory syndrome coronavi rus 2 (SARS-CoV-2). A high number of false positives (n=76) was obtained with Panther system (Hologic), but the number of false positives decreases as the relative light units (RLU) val ue increases. These results show that this technique can be a good option for sample screening but checking for positive results should be mandatory, especially those with low RLU values (AU)

Durante las múltiples oleadas de COVID-19 sufridas en todo el mundo, disponer de una prueba diagnóstica rápida y sensible se ha convertido en una prioridad para los laborato transcripción (TMA) AptimaTM SARS-CoV-2 que se ejecuta en el sistema Panther (Hologic) se presentó como una muy bue na opción para cubrir esta necesidad. Para evaluar este sis tema, se incluyeron en el estudio 570 muestras respiratorias y se procesaron tanto por el sistema Panther (Hologic) como por qRT-PCR (Thermo Fisher Science, Waltham, EE. UU.), téc nica utilizada actualmente para el diagnóstico del síndrome respiratorio agudo severo por coronavirus 2 (SARS-CoV-2). Se obtuvo un alto número de falsos positivos (n=76) con el sistema Panther (Hologic), pero el número de falsos positivos disminuye a medida que aumenta el valor de las unidades re lativas de luz (RLU). Estos resultados muestran que esta técnica puede ser una buena opción como técnica de screening, pero la verificación de resultados positivos debería ser obligatoria, especialmente aquellos con valores bajos de RLU (AU)

A Series of 69 COVID-related Dermatoses With Biopsy, Immunohistochemistry With Anti-spike 3, in situ Hybridization and PCR: A Critical Reappraisal of Viral Involvement in COVID-19 Skin Lesions / Serie de 69 dermatosis relacionadas con la COVID con biopsia, estudio inmunohistoquímico con anti-spike 3, hibridación in situ y PCR: Una revisión crítica de la afectación cutánea relacionada con la COVID-19

Background Despite the large number of articles published on skin lesions related to COVID-19, clinicopathological correlation has not been performed consistently and immunohistochemistry to demonstrate spike 3 protein expression has not been validated through RT-PCR. Material and method We compiled 69 cases of patients with confirmed COVID-19, where skin lesions were clinically and histopathologically studied. Immunohistochemistry (IHC) and RT-PCR was performed in skin biopsies. Results After a careful review of the cases, 15 were found to be dermatosis not related to COVID-19, while the rest of the lesions could be classified according to their clinical characteristics as vesicular (4), maculopapular eruptions (41), urticariform (9), livedo and necrosis (10) and pernio-like (5). Although histopathological features were similar to previously reported results, we found two previously unreported findings, maculopapular eruptions with squamous eccrine syringometaplasia and neutrophilic epitheliotropism. IHC showed in some cases endothelial and epidermal staining but RT-PCR was negative in all the tested cases. Thus, direct viral involvement could not be demonstrated. Conclusions Despite presenting the largest series of confirmed COVID-19 patients with histopathologically studied skin manifestations, direct viral involvement was difficult to establish. Vasculopathic and urticariform lesions seem to be those more clearly related to the viral infection, despite IHC or RT-PCR negative results failed to demonstrate viral presence. These findings, as in other dermatological areas, highlight the need of a clinico-pathological correlation to increase knowledge about viral involvement in COVID-19 skin-related lesions (AU)

Antecedentes A pesar del gran número de artículos publicados sobre las lesiones cutáneas relacionadas con la COVID-19, no se ha realizado una correlación clinicopatológica de manera consistente, y no ha sido validado el estudio de inmunohistoquímica para demostrar la expresión de la proteína spike 3 mediante RT-PCR. Material y métodos Recopilamos 69 casos de pacientes con COVID-19 confirmada, en los que se estudiaron las lesiones cutáneas a nivel clínico e histopatológico, habiéndose realizado la prueba inmunohistoquímica (IHQ) y RT-PCR en las biopsias cutáneas. Resultados Tras una revisión detallada de los casos, en 15 de ellos se encontró que la dermatosis no guardaba relación con la COVID-19, mientras que el resto de las lesiones podrían clasificarse de acuerdo con sus características clínicas como vesiculares (4), erupciones maculopapulares (41), urticariformes (9), livedo y necrosis (10) y de tipo perniosis (5). Aunque las características histopatológicas fueron similares a los resultados previamente reportados, encontramos dos hallazgos no reportados previamente: erupciones maculopapulares con siringometaplasia ecrina escamosa y epiteliotropismo neutrofílico. La IHQ reflejó en ciertos casos tinción endotelial y epidérmica, pero la prueba RT-PCR fue negativa en todos los casos probados. Por ello no pudo demostrarse el compromiso viral directo. Conclusiones A pesar de presentar la mayor serie de pacientes con COVID-19 confirmada y manifestaciones cutáneas histopatológicamente estudiadas, el compromiso viral directo fue difícil de establecer. Las lesiones vasculopáticas e urticariformes parecen ser las más claramente relacionadas con la infección viral, a pesar de que los resultados negativos de la IHQ o RT-PCR no pudieron demostrar la presencia viral (AU)

[Articulo traducido] Serie de 69 dermatosis relacionadas con la COVID-19 con biopsia, estudio inmunohistoquímico con anti-spike 3, hibridación in situ y PCR: Una revisión crítica de la afectación cutánea relacionada con la COVID-19 / A Series of 69 COVID-related Dermatoses With Biopsy, Immunohistochemistry With Anti-spike 3, in situ Hybridization and PCR: A Critical Reappraisal of Viral Involvement in COVID-19 Skin Lesions

Antecedentes A pesar del gran número de artículos publicados sobre las lesiones cutáneas relacionadas con la COVID-19, no se ha realizado una correlación clinicopatológica de manera consistente, y no ha sido validado el estudio de inmunohistoquímica para demostrar la expresión de la proteína spike 3 mediante RT-PCR. Material y métodos Recopilamos 69 casos de pacientes con COVID-19 confirmada, en los que se estudiaron las lesiones cutáneas a nivel clínico e histopatológico, habiéndose realizado la prueba inmunohistoquímica (IHQ) y RT-PCR en las biopsias cutáneas. Resultados Tras una revisión detallada de los casos, en 15 de ellos se encontró que la dermatosis no guardaba relación con la COVID-19, mientras que el resto de las lesiones podrían clasificarse de acuerdo con sus características clínicas como vesiculares (4), erupciones maculopapulares (41), urticariformes (9), livedo y necrosis (10) y de tipo perniosis (5). Aunque las características histopatológicas fueron similares a los resultados previamente reportados, encontramos dos hallazgos no reportados previamente: erupciones maculopapulares con siringometaplasia ecrina escamosa y epiteliotropismo neutrofílico. La IHQ reflejó en ciertos casos tinción endotelial y epidérmica, pero la prueba RT-PCR fue negativa en todos los casos probados. Por ello no pudo demostrarse el compromiso viral directo. Conclusiones A pesar de presentar la mayor serie de pacientes con COVID-19 confirmada y manifestaciones cutáneas histopatológicamente estudiadas, el compromiso viral directo fue difícil de establecer. Las lesiones vasculopáticas e urticariformes parecen ser las más claramente relacionadas con la infección viral, a pesar de que los resultados negativos de la IHQ o RT-PCR no pudieron demostrar la presencia viral (AU)

Background Despite the large number of articles published on skin lesions related to COVID-19, clinicopathological correlation has not been performed consistently and immunohistochemistry to demonstrate spike 3 protein expression has not been validated through RT-PCR. Material and method We compiled 69 cases of patients with confirmed COVID-19, where skin lesions were clinically and histopathologically studied. Immunohistochemistry (IHC) and RT-PCR was performed in skin biopsies. Results After a careful review of the cases, 15 were found to be dermatosis not related to COVID-19, while the rest of the lesions could be classified according to their clinical characteristics as vesicular (4), maculopapular eruptions (41), urticariform (9), livedo and necrosis (10) and pernio-like (5). Although histopathological features were similar to previously reported results, we found two previously unreported findings, maculopapular eruptions with squamous eccrine syringometaplasia and neutrophilic epitheliotropism. IHC showed in some cases endothelial and epidermal staining but RT-PCR was negative in all the tested cases. Thus, direct viral involvement could not be demonstrated. Conclusions Despite presenting the largest series of confirmed COVID-19 patients with histopathologically studied skin manifestations, direct viral involvement was difficult to establish. Vasculopathic and urticariform lesions seem to be those more clearly related to the viral infection, despite IHC or RT-PCR negative results failed to demonstrate viral presence. These findings, as in other dermatological areas, highlight the need of a clinico-pathological correlation to increase knowledge about viral involvement in COVID-19 skin-related lesions (AU)

TC torácica de baja dosis de radiación como técnica de cribado preoperatorio de infección por SARS-CoV-2 / Low-dose chest CT for preoperative screening for SARS-CoV-2 infection

Introducción: El objetivo del presente estudio fue evaluar el papel de la tomografía computarizada (TC) de tórax complementaria a la prueba de la reacción en cadena de la polimerasa con transcripción inversa (RT-PCR) en pacientes asintomáticos candidatos a cirugía electiva en el contexto de la pandemia de COVID-19. Material y métodos: Se incluyeron, de forma prospectiva, 464 pacientes asintomáticos que se sometieron a una triple estrategia de cribado de infección por SARS-CoV-2 (cuestionario de salud, prueba de RT-PCR y TC torácica de baja dosis) durante las 48 horas previas a la realización de una cirugía electiva. Un resultado positivo en la prueba de RT-PCR y/o la identificación de hallazgos tomográficos sugestivos de neumonía COVID-19 (categorías CO-RADS 4 y 5) fueron considerados criterios diagnósticos de infección por SARS-CoV-2. Resultados: La mayor parte de los pacientes se sometieron a cirugías de otorrinolaringología (64,7%). Ningún paciente presentó un resultado positivo en la prueba de RT-PCR ni síntomas sugestivos de infección por SARS-CoV-2 en el cuestionario de salud. Únicamente 22 (4,7%) mostraron signos compatibles con infección pulmonar; 20 de ellos atípica o indeterminada para COVID-19 (CO-RADS 2, 3) y 2 compatibles con neumonía COVID-19 en resolución. Durante el postoperatorio inmediato no se confirmó ningún caso positivo para SARS-CoV-2. Conclusión: En nuestra serie, la realización de una TC torácica de baja dosis de radiación en pacientes asintomáticos para el cribado preoperatorio de infección por SARS-CoV-2 no proporcionó un valor diagnóstico adicional a la RT-PCR y el cuestionario de salud.(AU)

Introduction: This study aimed to evaluate the role of chest computed tomography (CT) in complementing reverse transcription polymerase chain reaction (RT-PCR) in asymptomatic candidates for elective surgery in the context of the COVID-19 pandemic. Material and methods: We prospectively included 464 asymptomatic patients who underwent a triple screening workup for SARS-CoV-2 infection (health questionnaire, RT-PCR, and low-dose chest CT) during the 48hours prior to undergoing elective surgery. A positive RT-PCR and/or CT findings suggestive of COVID-19 (CO-RADS 4 / 5) were considered diagnostic criteria for SARS-CoV-2 infection. Results: Most patients (64.7%) underwent otorhinolaryngology surgery. No patients had positive RT-PCR results or symptoms suggestive of SARS-CoV-2 in the health questionnaire. Only 22 (4.7%) had signs compatible with lung infection; in 20 of these, the CT findings were atypical or indeterminate for COVID-19 (CO-RADS 2 / 3) and in 2 they were compatible with COVID-19 pneumonia in resolution. In the immediate postoperative period, no cases of SARS-CoV-2 infection were confirmed. Conclusion: In our series of asymptomatic patients, low-dose CT did not add any value to the results of RT-PCR and a health questionnaire in preoperative screening for SARS-CoV-2.(AU)

Serum microRNAs Catalog Asthma Patients by Phenotype

Background: Asthma is a chronic inflammatory condition of the airways with a complex pathophysiology. Stratification of asthma subtypes into phenotypes and endotypes should move the field forward, making treatment more effective and personalized. Eosinophils are the key inflammatory cells involved in severe eosinophilic asthma. Given the health threat posed by eosinophilic asthma, there is a need for reliable biomarkers to identify affected patients and treat them properly with novel biologics. microRNAs (miRNAs) are a promising diagnostic tool. Objective: The aim of this study was to identify serum miRNAs that can phenotype asthma patients. Methods: Serum miRNAs of patients with eosinophilic asthma (N=40) and patients with noneosinophilic asthma (N=36) were evaluated using next-generation sequencing, specifically miRNAs-seq, and selected miRNAs were validated using RT-qPCR. Pathway enrichment analysis of deregulated miRNAs was performed. Results: Next-generation sequencing revealed 15 miRNAs that were expressed differentially between eosinophilic and noneosinophilic asthma patients, although no differences were observed in the miRNome between atopic and nonatopic asthma patients. Of the 15 miRNAs expressed differentially between eosinophilic and noneosinophilic asthma patients, hsa-miR-26a-1-3p and hsa-miR-376a-3p were validated by RT-qPCR. Expression levels of these 2 miRNAs were higher in eosinophilic than in noneosinophilic asthma patients. Furthermore, expression values of hsa-miR-26a-1-3p correlated inversely with peripheral blood eosinophil count, and hsa-miR-376a-3p expression values correlated with FeNO values and the number of exacerbations. Additionally, in silico pathway enrichment analysis revealed that these 2 miRNAs regulate signaling pathways associated with the pathogenesis of asthma. Conclusion: hsa-miR-26a-1-3p and hsa-miR-376a-3p could be used to differentiate between eosinophilic and noneosinophilic asthma (AU)

Diagnóstico de SARS-CoV-2 mediante test de antígeno en un Servicio de Urgencias Pediátricas / Diagnosis of SARS-CoV-2 by antigen test in a Pediatric Emergency Service

Introducción. Encontrar un método diagnóstico para laenfermedad por SARS-CoV-2, eficiente y accesible ha sidouno de los grandes problemas a lo largo de la epidemia porCOVID 19.Pacientes y métodos. Estudio descriptivo retrospectivode los datos clínicos de los pacientes sometidos a test de antígenos para SARS-CoV-2 realizados en el Servicio de Urgencias Pediátricas de un hospital terciario, entre el 22/12/2021y el 10/02/2022, y su concordancia con el resultado de laRT-PCR de SARS-CoV-2 en caso de disponer de ésta.Resultados. Se realizaron 653 test de antígenos (53,9%varones), siendo positivos el 26,6%. La edad media fueestadísticamente mayor en aquellos con resultado positivo(67,3±51 meses, frente a a 51,95±51,3 meses). El síntomamás frecuente entre los pacientes positivos fue la fiebre en el79%. Entre los 387 pacientes con test negativo, se realizaron92 RT-PCR, resultando positivas 11 de ellas (12%) 9 de los 11pacientes con RT-PCR positivo tenían un contacto familiarestrecho y, de estas, 7 presentaban fiebre. Resulto significativa la relación entre tener un contacto familiar y un test deantígeno positivo (p<0,01), pero no con otro tipo de contacto.Discusión. Los pacientes que presentaron RT-PCR paraSARS-CoV-2 positiva, con test de antígeno negativo presentaban en su mayoría contacto familiar y fiebre. El contacto nofamiliar no tenía mayor porcentaje de falsos negativos queaquellos sin contacto conocido. La variación de positividadpuede deberse a las diferencias en la valoración de caso sospechoso y a la técnica de obtención de muestra. (AU)

Introduction. Finding an efficient and accessible diagnostic method for SARS-CoV-2 disease has been one of thebig problems throughout the COVID 19 epidemic.Patients and methods. Retrospective descriptive studyof the clinical data of patients undergoing antigen testsfor SARS-CoV-2 carried out in the Pediatric EmergencyDepartment of a tertiary hospital, between 22/12/2021and 10/02/2022, and its concordance with the result of theRT-PCR of SARS-CoV-2 if available.Result. 653 antigen tests were performed (53.9% male),26.6% were positive. The mean age was statistically higherin those with a positive result (67.3±51 months, comparedto 51.95±51.3 months). The most frequent symptom amongpositive patients was fever in 79%. Among the 387 patientswith a negative test, 92 RT-PCR were performed, resultingin positive 11 of them (12%) 9 of the 11 patients with positive RT-PCR had a close family contact and, of these, 7 hadfever. The relationship between having a family contact anda positive antigen test (p<0.01) was significant, but not with another type of contact. Discussion. Patients who presented RT-PCR for SARSCoV-2 positive, with negative antigen test had mostly familycontact and fever. Non-family contact had no higher percentage of false negatives than those with no known contact. The variation in positivity may be due to differencesin the assessment of suspected case and sample collection technique. (AU)

microRNA-377-3p inhibits osteosarcoma progression by targeting CUL1 and regulating Wnt/B-catenin signaling pathway

Objective Emerging studies highlight the crucial effects of microRNAs on cancer initiation and malignant progression of various tumors. This study focused on the biological effect of miR-377-3p on CUL1 and epithelial–mesenchymal transition (EMT) and Wnt/β-catenin pathways in osteosarcoma (OS). Methods We performed quantitative real-time polymerase chain reaction (qRT-PCR) to analyze miR-377-3p and CUL1 expression levels in OS tissues and MG-63 cells. Then, cell counting kit (CCK)-8 and Transwell assay were used to examine the functions of miR-377-3p in OS cell growth and metastasis abilities. Meanwhile, luciferase reporter assay was used to validate CUL1 as direct target of miR-377-3p. qRT-PCR and Western blot were then carried out to detect the impact of miR-377-3p on EMT and Wnt/β-catenin pathways. Tumor xenograft models were established to further examine the effects of miR-377-3p on OS tumorigenesis in vivo. Results miR-377-3p downregulation was frequently identified in OS tissues and cells, which was associated with worse prognosis of OS patients. Functional experiments showed miR-377-3p restoration could dramatically repress OS cell growth and migration by regulation of EMT and Wnt/β-catenin pathways. Moreover, luciferase reporter assay revealed that CUL1 acted as a functional target of miR-377-3p. Additionally, the elevated CUL1 expressions in OS tissues also indicated poor prognosis of OS patients. Furthermore, the OS tumor growth was also obviously inhibited by miR-377-3p overexpression in vivo. Conclusions Collectively, all the above findings revealed that miR-377-3p exerted anti-OS functions via CUL1 and EMT and Wnt/β-catenin pathways. These results may contribute to the development of clinical OS treatment (AU)

Aspectos básicos de la COVID-19 para el manejo desde atención primaria / Basic aspects of COVID-19 for management from primary care

El SARS-CoV-2 se transmite de persona a persona por inhalación o por el contacto con gotas y aerosoles respiratorios. El período de incubación mediano es 5,1 días. Fiebre, tos seca, disnea y fatiga, son los síntomas más comunes. Casi la mitad de los casos son asintomáticos. El espectro de la enfermedad varía desde cuadros leves (81%) a críticos (5%). La edad avanzada, el sexo masculino y las comorbilidades impactan negativamente sobre la gravedad y la mortalidad de la COVID-19. El diagnóstico de la patología aguda se realiza con pruebas de reacción en cadena de la polimerasa con transcriptasa inversa (RT-PCR) o de detección antigénica. En pacientes hospitalizados, el remdesivir reduce el tiempo de recuperación. Los corticoides orales se recomiendan en casos graves o críticos que requieren oxigenoterapia o ventilación mecánica. La tromboprofilaxis se sugiere en todos los casos graves y no graves con riesgo trombótico alto. La antibioterapia se limita a pacientes de alta sospecha de sobreinfección bacteriana. Los casos leves-moderados y graves tras el alta hospitalaria deben monitorizarse clínicamente durante un período mínimo de dos semanas.(AU)

SARS-CoV-2 is transmitted from person to person by inhalation or contact with respiratory droplets and aerosols. The median incubation period is 5.1 days. Fever, dry cough, dyspnea and fatigue are the most common symptoms. Almost half of the cases are asymptomatic. The spectrum of disease varies from mild (81%) to critical (5%). Older age, male gender and comorbidities negatively impact on the severity and mortality of COVID-19. The diagnosis of acute COVID-19 is made with RT-PCR or antigenic detection tests. In hospital patients, remdesivir reduces recovery time. Oral steroids are recommended for severe or critical cases requiring oxygen therapy or mechanical ventilation. Thromboprophylaxis is recommended in all severe and non-severe cases with high thrombotic risk. Antibiotherapy is limited to cases of high suspicion of bacterial superinfection. Mild-moderate and severe cases after discharge from hospital should be clinically monitored for a minimum period of two weeks.(AU)

Low prevalence of asthma in Mexican children and adults with a positive rtRT-PCR test for SARS-CoV-2: a cross-sectional study during the 2020 pandemic

Background: It has recently been argued that asthma does not increase the risk of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection. If so, the prevalence of asthma in subjects diagnosed with COVID-19 should be lower than in the general population. Objective: To determine the prevalence of asthma in Mexican children and adults with SARS-CoV-2 infection. Methods: A public database of the Epidemiological Surveillance System for Viral Respiratory Disease in Mexico was analyzed. Those who underwent the real-time reverse transcriptase-polymerase chain reaction-SARS-CoV-2 (rtRT-PCR-SARS-CoV-2) test from February 27 to June 21, 2020, were included. In addition to the prevalence of asthma, some factors associated with it were investigated. Results: Data from 417,366 subjects were analyzed. Asthma prevalence in children, adults, and global were 3.7%, 3.3%, and 3.3%, respectively. Although the asthma prevalence was lower in SARS-CoV-2 positive over negative patients, significant differences were only found in adults (2.8% vs. 3.7% respectively; odds ratio (OR) = 0.74; 95% confidence interval (CI): 0.71–0.77); but not in children (3.5% vs. 3.8%, respectively; OR = 0.91; 95%CI: 0.76–1.10). Multivariate analysis showed in younger than 18 years that girls and immunosuppression were factors associated with a decrease in the odds to develop asthma. In adults, asthma was positively associated with females, obesity, smoking, immunosuppression, chronic obstructive pulmonary disease, arterial hypertension, and cardiovascular disease. Conclusion: The prevalence of asthma in child and adult were lower than those previously reported. Our study seems to support the hypothesis that asthma patients have a lower risk of SARS-CoV-2 infection. Further studies are required to demonstrate the consistency of our findings (AU)

Clínica diferencial en niños infectados por SARS-CoV-2, trazabilidad de contactos y rentabilidad de pruebas diagnósticas: estudio observacional transversal / Differential clinic in children infected by SARS-CoV-2, traceability of contacts and cost-effectiveness of diagnostic tests: Cross-sectional observational study

INTRODUCCIÓN: Ante la posible coexistencia de la infección por el virus SARS-CoV-2 con otras infecciones estacionales, se pretende identificar síntomas diferenciales. Se ha estudiado el papel de los niños en el contagio intrafamiliar y la sensibilidad de la reacción en cadena de la polimerasa con transcriptasa inversa (RT-PCR) en un área con baja transmisión comunitaria. MATERIAL Y MÉTODOS: Estudio observacional transversal. Pacientes entre 0-15 años estudiados por técnica RT-PCR por sospecha clínica de infección por virus SARS-CoV-2 en los meses de marzo-mayo del 2020. Encuesta sobre síntomas y contactos. Determinación de anticuerpos anti-SARS-CoV-2 al menos 21 días después del test RT- PCR. RESULTADOS: Se incluyó a 126 pacientes, 33 con infección confirmada y edad media 8,4 años (IC del 95%, 6,8-10,0) superior a los no infectados. La fiebre fue el síntoma más común y con mayor sensibilidad. Las diferencias encontradas fueron una mayor frecuencia de anosmia (p = 0,029) y cefalea (p = 0,009) entre los niños infectados con una especificidad del 96,7 y el 81,5% respectivamente. No hubo diferencias en la duración de los síntomas. Un 81,8% de los infectados fue probablemente contagiado en el núcleo familiar, en un 85,2% por un progenitor que trabajaba fuera del hogar. La sensibilidad de RT-PCR fue 70,9% y su valor predictivo negativo 91,1%. CONCLUSIONES: El cuadro clínico es inespecífico y los síntomas más específicos difíciles de detectar en niños más pequeños. Los niños tuvieron un papel reducido en la transmisión intrafamiliar. La sensibilidad de la RT-PCR podría estar relacionada con una menor contagiosidad infantil tras una semana de infección

INTRODUCTION: Given the possible coexistence of infection by the SARS-CoV-2 with other seasonal infections, the aim is to identify differential symptoms. I know has studied the role of children in intrafamily contagion and the sensitivity of reverse transcriptase polymerase chain reaction (RT-PCR) in an area with low community transmission. MATERIAL AND METHODS: Cross-sectional observational study. Patients between 0-15 years studied by RT-PCR technique due to clinical suspicion of infection by SARS-CoV-2 virus in the months of March-May 2020. Survey on symptoms and contacts. Determination of Anti-SARS-CoV-2 antibodies at least 21 days after the RT-PCR test. RESULTS: 126 patients were included, 33 with confirmed infection and age mean 8.4 years (95% CI 6.8-10,5) higher than not infected. Fever was the most common and with greater sensitivity. The differences found were a greater frequency of anosmia (P = 0.029) and headache (P = .009) among children infected with a specificity of 96.7% and 81.5% respectively. There were no differences in the duration of the symptoms. 81.8% of those infected were probably infected in the nucleus 85.2% by a parent who worked outside the home. The sensitivity of RT-PCR was 70.9% and its negative predictive value 91.1%. CONCLUSIONS: The clinical picture is nonspecific and the symptoms more specific difficult to detect in younger children. Children had a reduced role in the intrafamily transmission. The sensitivity of RT-PCR could be related to a less contagiousness in children after one week of infection

Evaluation of color Doppler ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of breast cancer

Purpose To evaluate the clinical value of color Doppler ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of breast cancer (BC). Methods 129 patients with BC and 50 patients with benign breast lesions were selected. They were scanned by color Doppler ultrasound on the breast. The relative expression of miR-21 and miR-27a in plasma was detected by real-time fluorescence quantitative PCR. The clinical value of color Doppler ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of BC was analyzed retrospectively. Results Two-dimensional ultrasound images of BC showed irregular shape, unclear boundary, burr-like edge, uneven internal echo, weakened rear echo, microcalcification, aspect ratio > 1 and axillary lymph node metastasis. The grade of blood flow signal was improved, mainly in grade II and above, which was statistically different from that of benign control group (P < 0.001). The relative expression of plasma miR-21 and miR-27a in breast cancer group was significantly higher than that in benign control group (P < 0.001). The relative expressions of plasma miR-21 and miR-27a in breast cancer group were significantly correlated with histological grade, clinical stage, lymph node metastasis and postoperation. The relative expression levels of miR-21 and miR-27a in plasma of patients with low histological grade (grade III), high clinical stage (stage III, IV) and lymph node metastasis before operation were significantly higher than those in patients with high histological grade (grade I, II), low clinical stage (stage I, II) and no lymph node metastasis after operation (P < 0.01) (AU)

Noncoding RNAs as potential biomarkers for DIPG diagnosis and prognosis: XIST and XIST-210 involvement

Purpose Diffuse intrinsic pontine gliomas (DIPGs) are the most fatal primary brainstem tumors in pediatric patients. The identification of new molecular features, mediating their formation and progression, as non-coding RNAs (ncRNAs), would be of great importance for the development of effective treatments. Methods We analyzed the DIPGs transcriptome with the HTA2.0 array and it was compared with pediatric non-brainstem astrocytoma expression profiles (GSE72269). Results More than 50% of the differentially expressed transcripts were ncRNAs and based on this, we proposed a DIPGs ncRNA signature. LncRNAs XIST and XIST-210, and the HBII-52 and HBII-85 snoRNA clusters were markedly downregulated in DIPGs. qPCR assays demonstrated XIST downregulation in all non-brainstem astrocytomas, in a gender, age, and brain location-independent manner, as well as in DIPGs affecting boys; however, DIPGs affecting girls showed both downregulation and upregulation of XIST. Girls’ with longer survival positively correlated with XIST expression. Conclusions The involvement of ncRNAs in DIPGs is imminent and their expression profile is useful to differentiate them from non-neoplastic tissues and non-brain stem astrocytomas, which suggests their potential use as DIPG biomarkers. In fact, XIST and XIST-210 are potential DIPG prognostic biomarkers (AU)

CLIC1 knockout inhibits invasion and migration of gastric cancer by upregulating AMOT-p130 expression

Purpose To explore the regulatory relationship between Chloride intracellular channel 1 (CLIC1) and Angiomotin (AMOT)-p130, and reveal the role of AMOT-p130 in gastric cancer (GC). Methods Immunohistochemistry was performed to analyze the expression of CLIC1 and AMOT-p130 in GC tissues and adjacent tissues. The expression of AMOT-p130 upon CLIC1 silencing was analyzed using RT-PCR, western blot, and immunofluorescence in GC cells. Transwell and wound-healing assays were performed to detect migration and invasion in GC cells. The changes in EMT-related proteins were detected using western blot. Results Our study found that high CLIC1 expression was significantly associated with low AMOT-p130 expression in GC tissues. Silencing CLIC1 expression in MGC-803 cells (MGC-803 CLIC1 KO) and AGS cells (AGS CLIC1 KO) decreased the invasive and migratory abilities of tumor cells, which were induced by the upregulation of AMOT-p130. Subsequently, we demonstrated that AMOT-p130 inhibits the invasive and migratory abilities of GC cells by inhibiting epithelial–mesenchymal transition. Conclusions Our study suggests that AMOT-p130 could inhibit epithelial–mesenchymal transition in GC cells. CLIC1 may participate in the metastatic progression of GC by downregulating the expression of AMOT-p130 (AU)

Utilidad de la sangre seca del talón (tarjeta de Guthrie) en el diagnóstico de infección congénita por enterovirus / Use of dried blood from the heel (Guthrie card) in the diagnosis of congenital enterovirus infection

No disponible

Multiple Reversed Halo Sign on Chest CT in COVID-19 Pneumonia / Múltiples signos del halo invertido en la TC de tórax de una neumonía por COVID-19

No disponible

Características de los pacientes con enfermedad neumocócica invasora que requieren ingreso en la unidad de cuidados intensivos / Characteristics of patients with invasive pneumococcal disease requiring admission to intensive care units

INTRODUCCIÓN: La enfermedad neumocócica invasora (ENI) puede requerir ingreso en la unidad de cuidados intensivos pediátricos (UCIP). El objetivo de este trabajo es analizar las características epidemiológicas, clínicas y microbiológicas asociadas a la ENI que predisponen el ingreso en la UCIP. MATERIAL Y MÉTODOS: Estudio prospectivo de casos diagnosticados con ENI en tres hospitales pediátricos de Barcelona entre enero de 2012 y junio de 2016. Se analizaron las asociaciones entre el ingreso en la UCIP y las variables epidemiológicas, clínicas y microbiológicas. RESULTADOS: Se incluyeron 263 casos con ENI. El 19% (n = 50) requirió ingreso en la UCIP. El 100% (7) de los pacientes con shock séptico, 84,2% (16) con meningitis y 15,2% (23) con neumonía complicada ingresaron en la UCIP. Las complicaciones más frecuentes fueron pulmonares (35,2%) y neurológicas (39,5%). La razón entre ingreso y no ingreso en la UCIP fue 4,17 veces mayor en los sujetos con enfermedad de base. Los serotipos asociados al ingreso en la UCIP fueron el 19A (23% del total de este serotipo), el 14 (20%), el 3 (17%) y el serotipo 1 (12,5%). CONCLUSIONES: La ENI requiere ingreso en la UCIP en caso de shock séptico y meningitis, no así, de entrada, la neumonía complicada. El porcentaje de ingresos es mayor en los niños con enfermedad de base. El ingreso en la UCIP conlleva una estancia más prolongada, así como complicaciones durante la fase aguda y secuelas, sobre todo, neurológicas. Los serotipos de los pacientes que ingresaron en la UCIP fueron, predominantemente, serotipos vacunales

INTRODUCTION: Patients with invasive pneumococcal disease (IPD) may require admission into paediatric intensive care units (PICU). The aim of this study is to analyse the epidemiological, clinical, and microbiological characteristics associated with IPD that may require admission to the PICU. MATERIAL AND METHODS: A prospective study was conducted on cases of IPD diagnosed in three Paediatric Hospitals in Barcelona between January 2012 and June 2016. An analysis was made of the associations between the admission to PICU and the epidemiological, clinical, and microbiological variables. RESULTS: A total of 263 cases with IPD were included, of which 19% (n = 50) required admission to PICU. Patients with septic shock (7; 100%), meningitis (16; 84.2%), and those with complicated pneumonia (23; 15.2%) were admitted to the PICU. The most frequent complications were pulmonary (35.2%) and neurological (39.5%). The ratio between admission and non-admission to PICU was 4.7 times higher in subjects with an underlying disease. The serotypes associated with PICU admission were 19A (23% of the total of this serotype), serotype 14 (20%), serotype 3 (17%), and serotype 1 (12.5%). CONCLUSIONS: IPD required PICU admission in cases of septic shock and meningitis, and less so with complicated pneumonia. The percentage of admissions is greater in children with an underlying disease. Admission into the PICU involves a longer stay, complications during the acute phase, as well as sequelae, particularly neurological ones. The serotypes of the patients that were admitted to PICU were predominantly vaccine serotypes