OBJECTIVES:To quantify immunological dysfunction in surgical patients with presence/absence of sepsis using a droplet digital polymerase chain reaction (ddPCR) transcriptomic analysis. The study also ...aims to evaluate this approach for improving identification of sepsis in these patients.
BACKGROUND:Immune dysregulation is a central event in sepsis. Quantification of the expression of immunological genes participating in the pathogenesis of sepsis could represent a new avenue to improve its diagnosis.
METHODS:Expression of 6 neutrophil protease genes (MMP8, OLFM4, LCN2/NGAL, LTF, PRTN3, MPO) and also of 5 genes involved in the immunological synapse (HLA-DRA, CD40LG, CD3E, CD28, ICOS) was quantified in blood from 101 surgical patients with sepsis, 53 uninfected surgical patients, and 16 blood donors by using ddPCR. Areas under receiver operating characteristic curves (AUROC) and multivariate regression analysis were employed to test individual genes and gene ratios to identify sepsis, in comparison with procalcitonin.
RESULTS:Sepsis-induced overexpression of neutrophil protease genes and depressed expression of immunological synapse genes. MMP8/HLA-DRA, LCN2/HLA-DRA outperformed procalcitonin in differentiating between patients with sepsis and surgical controls in the AUROC analysisLCN2/HLA-DRA0.90 (0.85–0.96), MMP8/HLA-DRA0.89 (0.84–0.95), procalcitonin0.80 (0.73–0.88) (AUROC, confidence interval 95%), and also in the multivariate analysisLCN2/HLA-DRA8.57 (2.25–32.62); MMP8/HLA-DRA8.03 (2.10–30.76), procalcitonin4.20 (1.15–15.43) odds ratio (confidence interval 95%). Gene expression levels of HLA-DRA were an independent marker of hospital mortality.
CONCLUSIONS:Quantifying the transcriptomic ratios MMP8/HLA-DRA, LCN2/HLA-DRA by ddPCR is a promising approach to improve sepsis diagnosis in surgical patients.
Early recognition of sepsis is a key factor to improve survival to this disease in surgical patients, since it allows prompt control of the infectious source. Combining pro-inflammatory and ...immunosupression biomarkers could represent a good strategy to improve sepsis detection. Here we evaluated the combination of procalcitonin (PCT) with gene expression levels of HLA-DRA to detect sepsis in a cohort of 154 surgical patients (101 with sepsis and 53 with no infection). HLA-DRA expression was quantified using droplet digital PCR, a next-generation PCR technology. Area under the receiver operating curve analysis (AUROC) showed that the PCT/HLA-DRA ratio outperformed PCT to detect sepsis (AUROC CI95%, p): PCT: 0.80 0.73-0.88, <0.001; PCT/HLA-DRA: 0.85 0.78-0.91, <0.001. In the multivariate analysis, the ratio showed a superior ability to predict sepsis compared to that of PCT (OR CI 95%, p): PCT/HLA-DRA: 7.66 1.82-32.29, 0.006; PCT: 4.21 1.15-15.43 0.030. Multivariate analysis was confirmed using a new surgical cohort with 74 sepsis patients and 21 controls: PCT/HLA-DRA: 34.86 1.22-995.08, 0.038; PCT: 5.52 0.40-75.78, 0.201. In conclusion, the combination of PCT with HLA-DRA is a promising strategy for improving sepsis detection in surgical patients.
Summary Objectives Sepsis is characterised by the frequent presence of organ failure and marked immunologic alterations. We studied the association between the extent of organ failure and the ...transcriptomic response of septic patients. Methods Gene expression profiles in the blood of 74 surgical patients with sepsis were compared with those of 30 surgical patients with no sepsis. Differentially expressed genes were assessed for their correlation with the sequential organ failure (SOFA) score. Results The expression levels of a group of genes participating in the cell cycle (HIST1H1C, CKS2, CCNA2, CDK1, CCNB2, CIT, CCNB1, AURKA, RAD51), neutrophil protease activity (ELANE, ADORA3, MPO, MMP8, CTSG), IL-1R and IL-18R response correlated directly with SOFA and mortality. Genes involved in T cell (LCK, CD3G, CD3D, ZAP70, ICOS, CD3E, CD28, IL2RB, CD8B, CD8A, CD40LG, IL23A, CCL5, SH2D1A, ITK, CD247, TBX21, GATA3, CCR7, LEF1, STAT4) and NK cell immunity (CD244, KLRK1, KLRD1) were inversely associated with SOFA and mortality. Conclusions The extent of organ failure in sepsis correlates directly with the existence of imbalanced innate and adaptive responses at the transcriptomic level. Quantification of the expression levels of the genes identified here could contribute to the simultaneous assessment of disease severity and immunological alterations in sepsis.
Nowadays, mortality rates in intensive care units are the highest of all hospital units. However, there is not a reliable prognostic system to predict the likelihood of death in patients with ...postsurgical shock. Thus, the aim of the present work is to obtain a gene expression signature to distinguish the low and high risk of death in postsurgical shock patients. In this sense, mRNA levels were evaluated by microarray on a discovery cohort to select the most differentially expressed genes between surviving and non-surviving groups 30 days after the operation. Selected genes were evaluated by quantitative real-time polymerase chain reaction (qPCR) in a validation cohort to validate the reliability of data. A receiver-operating characteristic analysis with the area under the curve was performed to quantify the sensitivity and specificity for gene expression levels, which were compared with predictions by established risk scales, such as acute physiology and chronic health evaluation (APACHE) and sequential organ failure assessment (SOFA).
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genes were upregulated in the non-surviving group of the discovery cohort, and their predictive power was confirmed in the validation cohort. This work offers new biomarkers based on transcriptional patterns to classify the postsurgical shock patients according to low and high risk of death. The results present more accuracy than other mortality risk scores.
•Gene expression patterns can distinguish septic shock from non-septic shock.•Differentially expressed genes were IGHG1, IL1R2, LCN2, LTF, MMP8, and OLFM4.•This tool allows a correct diagnosis to ...provide an immediate and specific treatment.
To obtain a gene expression signature to distinguish between septic shock and non-septic shock in postoperative patients, since patients with both conditions show similar signs and symptoms.
Differentially expressed genes were selected by microarray analysis in the discovery cohort. These genes were evaluated by quantitative real time polymerase chain reactions in the validation cohort to determine their reliability and predictive capacity by receiver operating characteristic curve analysis.
Differentially expressed genes selected were IGHG1, IL1R2, LCN2, LTF, MMP8, and OLFM4. The multivariate regression model for gene expression presented an area under the curve value of 0.922. These genes were able to discern between both shock conditions better than other biomarkers used for diagnosis of these conditions, such as procalcitonin (0.589), C-reactive protein (0.705), or neutrophils (0.605).
Gene expression patterns provided a robust tool to distinguish septic shock from non-septic shock postsurgical patients and shows the potential to provide an immediate and specific treatment, avoiding the unnecessary use of broad-spectrum antibiotics and the development of antimicrobial resistance, secondary infections and increase health care costs.
El presente artículo investiga el saber que tiene el alumnado de secundaria sobre la inmunidad y las vacunas, para lo que se empleó un cuestionario de respuesta abierta junto a su rúbrica de ...evaluación. Así la rúbrica contempla cuatro categorías de respuesta para cada pregunta, cuyos significados revelan el tipo de concepción que mantiene el alumnado. Los resultados cualitativos obtenidos manifiestan ideas poco acordes al modelo escolar de inmunidad; no obstante, un análisis de Rasch muestra un saber articulado, el cual es evidenciado a través de la unidimensionalidad de las medidas obtenidas. Lo anteriormente mencionado sugiere la posibilidad de utilizar las mismas como guía de un posible itinerario de progresión que facilite el aprendizaje de los estudiantes a lo largo de la enseñanza. Estos resultados no solo tienen implicaciones a nivel teórico y de la práctica del aula, sino que también conllevan a la validación de la rúbrica construida para su uso con esta población de estudiantes.
This article investigates the knowledge of secondary school students about immunity and vaccines, using an open ended questionnaire along with its evaluation rubric. Thus, the rubric includes four response categories for each question, whose meanings reveal the kinds of ideas held by the students. The qualitative results obtained show ideas that are not very consistent with the referent learning model for immunity; however, an analysis of Rasch shows articulated knowledge, which is evidence through the unidimensionality of the measurements obtained. This suggests the possibility of using them as a guide to a possible progression itinerary that facilitates student learning over the course of their education. These results not only have implications at a theoretical and classroom practice levels, but also entail the validation of the rubric constructed for use with this student population.
Este artigo investiga o conhecimento de alunos do ensino médio sobre o a imunidade e as vacinas, utilizando um questionário de resposta aberta juntamente com a respetiva rubrica de avaliação. Assim, a rubrica contempla quatro categorias de resposta para cada questão, cujos significados revelam o tipo de concepção que os alunos possuem. Os resultados qualitativos obtidos mostram ideias que não condizem com o modelo escolar de imunidade; no entanto, uma análise Rasch mostra um conhecimento articulado, que é evidenciado pela unidimensionalidade das medidas obtidas. O que foi mencionado anteriormente, sugere a possibilidade de utilizá-los como guia para um possível roteiro de progressão que facilite o aprendizado dos alunos ao longo do ensino. Estes resultados não só têm implicações a nível teórico e prático em sala de aula, como também implicam à validação da rubrica construída para a utilização com esta população de alunos.
To analyze the incidence of Covid-19 in patients who are chronic users of hydroxychloroquine.
Cross-sectional retrospective observational multicenter study in health areas and districts from Castilla ...La-Mancha and Andalucia. Of the 4451 participants included in the first recruitment, 3817 with valid data were selected. The main variable of the study is the presence or absence of Covid-19 infection by clinical, serological or polymerase chain reaction diagnosis. Sociodemographic and clinical variables and treatment and concomitant comorbidities were recorded.
169 (4,45%) patients had Covid-19 infection, of which 12 (7.1%) died and 32 (18.9%) required hospital admission. Previous respiratory pathology was related to Covid-19 infection (P<.05). Maculopathy appears in 5.3% of patients and is significantly related to the dose of hydroxychloroquine consumed (P<.05).
There is no relationship between chronic use of hydroxychloroquine and the incidence of Covid-19.
En este trabajo se estudia el modelo de inmunidad y vacuna que presentan los libros de texto de educación secundaria, considerados un nexo entre el currículum y la práctica docente. El objetivo de ...nuestro estudio era explorar qué contenidos se incluyen en dichos modelos y cómo se articulan. A la vez, se propone un procedimiento basado en la elaboración de mapas conceptuales para alcanzar tal fin. En general, los modelos de los libros de texto de nuestro corpus de estudio presentan procesos celulares de la inmunidad centrados en lo individual antes que en lo colectivo. Las conclusiones de esta investigación suponen un diagnóstico parcial de cómo se aborda la enseñanza de la inmunología en las aulas para así diseñar un modelo que permita poner en práctica estrategias y recursos que impliquen al alumnado en la modelización de la inmunidad.
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