Fecal Microbiota of Toxigenic Clostridioides difficile-Associated Diarrhea

Clostridioides difficile infection (CDI) is currently one of the most important causes of infectious diarrhea in developed countries and the main cause in healthcare settings. Here, we characterized the gut microbiota from the feces of 57 patients with diarrhea from nosocomial and community-acquired CDI. We performed an ecological analysis by high-throughput sequencing of the V3-V4 region of 16S rRNA amplicons and evaluated the association of the various ecological profiles with CDI risk factors. Among all samples Bacteroidaceae 31.01%, Enterobacteriaceae 9.82%, Lachnospiraceae 9.33%, Tannerellaceae 6,16%, and Ruminococcaceae 5.64%, were the most abundant families. A reduced abundance of Bacteroides was associated with a poor CDI prognosis, with severe diarrhea and a high incidence of recurrence. This reduction was associated with a weakened host immune system and previous aggressive antibiotherapy. Peptostreptococcaceae family was 1.56% overall and within the family the only identified member was the genus Clostridioides, positively correlated with the presence of Akkermansia that may be predictive of the presence of a CDI. Finally, a relevant aspect that must be considered in clinical practice is the misdiagnosis of CDI, as patients with a stool sample that tests positive for C. difficile are usually diagnosed with CDI and subsequently treated as such. However, co-infection with other pathogenic agents often plays an important role in the development of diarrhea, and must be considered when prescribing antibiotic treatment

Th1 and Th17 hypercytokinemia as early host response signature in severe pandemic influenza

Abstract Introduction Human host immune response following infection with the new variant of A/H1N1 pandemic influenza virus (nvH1N1) is poorly understood. We utilize here systemic cytokine and antibody levels in evaluating differences in early immune response in both mild and severe patients infected with nvH1N1. Methods We profiled 29 cytokines and chemokines and evaluated the haemagglutination inhibition activity as quantitative and qualitative measurements of host immune responses in serum obtained during the first five days after symptoms onset, in two cohorts of nvH1N1 infected patients. Severe patients required hospitalization (n = 20), due to respiratory insufficiency (10 of them were admitted to the intensive care unit), while mild patients had exclusively flu-like symptoms (n = 15). A group of healthy donors was included as control (n = 15). Differences in levels of mediators between groups were assessed by using the non parametric U-Mann Whitney test. Association between variables was determined by calculating the Spearman correlation coefficient. Viral load was performed in serum by using real-time PCR targeting the neuraminidase gene. Results Increased levels of innate-immunity mediators (IP-10, MCP-1, MIP-1β), and the absence of anti-nvH1N1 antibodies, characterized the early response to nvH1N1 infection in both hospitalized and mild patients. High systemic levels of type-II interferon (IFN-γ) and also of a group of mediators involved in the development of T-helper 17 (IL-8, IL-9, IL-17, IL-6) and T-helper 1 (TNF-α, IL-15, IL-12p70) responses were exclusively found in hospitalized patients. IL-15, IL-12p70, IL-6 constituted a hallmark of critical illness in our study. A significant inverse association was found between IL-6, IL-8 and PaO2 in critical patients. Conclusions While infection with the nvH1N1 induces a typical innate response in both mild and severe patients, severe disease with respiratory involvement is characterized by early secretion of Th17 and Th1 cytokines usually associated with cell mediated immunity but also commonly linked to the pathogenesis of autoimmune/inflammatory diseases. The exact role of Th1 and Th17 mediators in the evolution of nvH1N1 mild and severe disease merits further investigation as to the detrimental or beneficial role these cytokines play in severe illness

Effect of viral storm in patients admitted to intensive care units with severe COVID-19 in Spain: a multicentre, prospective, cohort study

Background: The contribution of the virus to the pathogenesis of severe COVID-19 is still unclear. We aimed to evaluate associations between viral RNA load in plasma and host response, complications, and deaths in critically ill patients with COVID-19. Methods: We did a prospective cohort study across 23 hospitals in Spain. We included patients aged 18 years or older with laboratory-confirmed SARS-CoV-2 infection who were admitted to an intensive care unit between March 16, 2020, and Feb 27, 2021. RNA of the SARS-CoV-2 nucleocapsid region 1 (N1) was quantified in plasma samples collected from patients in the first 48 h following admission, using digital PCR. Patients were grouped on the basis of N1 quantity: VIR-N1-Zero ([removed]2747 N1 copies per mL). The primary outcome was all-cause death within 90 days after admission. We evaluated odds ratios (ORs) for the primary outcome between groups using a logistic regression analysis. Findings: 1068 patients met the inclusion criteria, of whom 117 had insufficient plasma samples and 115 had key information missing. 836 patients were included in the analysis, of whom 403 (48%) were in the VIR-N1-Low group, 283 (34%) were in the VIR-N1-Storm group, and 150 (18%) were in the VIR-N1-Zero group. Overall, patients in the VIR-N1-Storm group had the most severe disease: 266 (94%) of 283 patients received invasive mechanical ventilation (IMV), 116 (41%) developed acute kidney injury, 180 (65%) had secondary infections, and 148 (52%) died within 90 days. Patients in the VIR-N1-Zero group had the least severe disease: 81 (54%) of 150 received IMV, 34 (23%) developed acute kidney injury, 47 (32%) had secondary infections, and 26 (17%) died within 90 days (OR for death 0·30, 95% CI 0·16–0·55; p<0·0001, compared with the VIR-N1-Storm group). 106 (26%) of 403 patients in the VIR-N1-Low group died within 90 days (OR for death 0·39, 95% CI 0·26–0·57; p[removed]11 página

Viral RNA load in plasma is associated with critical illness and a dysregulated host response in COVID‑19

Background. COVID-19 can course with respiratory and extrapulmonary disease. SARS-CoV-2 RNA is detected in respiratory samples but also in blood, stool and urine. Severe COVID-19 is characterized by a dysregulated host response to this virus. We studied whether viral RNAemia or viral RNA load in plasma is associated with severe COVID-19 and also to this dysregulated response. Methods. A total of 250 patients with COVID-19 were recruited (50 outpatients, 100 hospitalized ward patients and 100 critically ill). Viral RNA detection and quantification in plasma was performed using droplet digital PCR, targeting the N1 and N2 regions of the SARS-CoV-2 nucleoprotein gene. The association between SARS-CoV-2 RNAemia and viral RNA load in plasma with severity was evaluated by multivariate logistic regression. Correlations between viral RNA load and biomarkers evidencing dysregulation of host response were evaluated by calculating the Spearman correlation coefficients. Results. The frequency of viral RNAemia was higher in the critically ill patients (78%) compared to ward patients (27%) and outpatients (2%) (p < 0.001). Critical patients had higher viral RNA loads in plasma than non-critically ill patients, with non-survivors showing the highest values. When outpatients and ward patients were compared, viral RNAemia did not show significant associations in the multivariate analysis. In contrast, when ward patients were compared with ICU patients, both viral RNAemia and viral RNA load in plasma were associated with critical illness (OR [CI 95%], p): RNAemia (3.92 [1.183–12.968], 0.025), viral RNA load (N1) (1.962 [1.244–3.096], 0.004); viral RNA load (N2) (2.229 [1.382–3.595], 0.001). Viral RNA load in plasma correlated with higher levels of chemokines (CXCL10, CCL2), biomarkers indicative of a systemic inflammatory response (IL-6, CRP, ferritin), activation of NK cells (IL-15), endothelial dysfunction (VCAM-1, angiopoietin-2, ICAM-1), coagulation activation (D-Dimer and INR), tissue damage (LDH, GPT), neutrophil response (neutrophils counts, myeloperoxidase, GM-CSF) and immunodepression (PD-L1, IL-10, lymphopenia and monocytopenia). Conclusions. SARS-CoV-2 RNAemia and viral RNA load in plasma are associated with critical illness in COVID-19. Viral RNA load in plasma correlates with key signatures of dysregulated host responses, suggesting a major role of uncontrolled viral replication in the pathogenesis of this disease.This work was supported by awards from the Canadian Institutes of Health Research, the Canadian 2019 Novel Coronavirus (COVID-19) Rapid Research Funding initiative (CIHR OV2 – 170357), Research Nova Scotia (DJK), Atlantic Genome/Genome Canada (DJK), Li-Ka Shing Foundation (DJK), Dalhousie Medical Research Foundation (DJK), the “Subvenciones de concesión directa para proyectos y programas de investigación del virus SARS‐CoV2, causante del COVID‐19”, FONDO–COVID19, Instituto de Salud Carlos III (COV20/00110, CIBERES, 06/06/0028), (AT) and fnally by the “Convocatoria extraordinaria y urgente de la Gerencia Regional de Salud de Castilla y León, para la fnanciación de proyectos de investigación en enfermedad COVID-19” (GRS COVID 53/A/20) (CA). DJK is a recipient of the Canada Research Chair in Translational Vaccinology and Infammation. APT was funded by the Sara Borrell Research Grant CD018/0123 funded by Instituto de Salud Carlos III and co-fnanced by the European Development Regional Fund (A Way to Achieve Europe programme). The funding sources did not play any role neither in the design of the study and collection, not in the analysis, in the interpretation of data or in writing the manuscript

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Quadrivalent cell culture influenza virus vaccine. Comparison to egg-derived vaccine

Influenza virus infections pose a serious public health problem and vaccination is the most effective public health intervention against them. The current manufacture of influenza vaccines in embryonated chicken eggs entails significant limitations. These limitations have been overcome by producing vaccines in cell culture, which allow a faster and more flexible response to potential pandemic threats. Given the impact of influenza B virus on disease burden, the availability of quadrivalent vaccines is useful for increasing the rate of protection from disease. This paper analyzes the limitations of the current production of influenza vaccine in eggs and the advantages of vaccines developed in cell culture, as well as their safety, tolerability, efficacy and effectiveness. Additionally, we reflect on the contribution of new quadrivalent vaccines from cell culture as an alternative in seasonal vaccination campaigns against influenza

La utilización de los indicadores bibliométricos para evaluar la actividad investigadora

The use of bibliometric indicators in research performance assessment. Research assessment is essential for the right management of the resources devoted to this activity since it allows to know the performance of the scientific activity as well as its impact in society. With the results of this evaluation, the public funds allocated to this activity are justified in the eyes of society. But evaluating research is hardly accurate because it is not an exact science. There are several criteria used to assess this activity for individuals as well as for research groups. Among these criteria, we find the number of scientific publications published in a period, together with the number of times that these publications are cited in other papers or books, the number of patents or copyrights, the recognition awarded to the authors of the publications (prizes, honours, etc.), or the capacity to obtain public and private funds for these activities. Nevertheless, from all of these criteria, bibliometric indicators or indexes are one of the most used tools, since they provide quantitative and qualitative (i.e. the impact) information on the scientific production.La evaluación de la investigación es necesaria para la gestión y planificación de los recursos destinados a la investigación ya que permite conocer el rendimiento de la actividad científica así como su impacto en la sociedad. Con los resultados de esta evaluación se justifican ante la sociedad las partidas presupuestarias destinadas a esta actividad. Pero evaluar es una tarea ardua e imposible en exactitud debido a que la actividad científica no es exacta. Se utilizan diversos criterios para evaluar dicha actividad tanto para investigadores a nivel individual como para grupos de investigación. Entre estos criterios se incluyen el número de publicaciones científicas producidas en un periodo dado, el número de veces que estas publicaciones son citadas en otros artículos o libros, el número de patentes o registros de propiedad intelectual, el reconocimiento otorgado a los autores de las publicaciones (premios, honores, etc...), y también la capacidad de captación de financiación tanto pública como privada para la realización de estas actividades. De todos estos criterios, los indicadores o índices bibliométricos son una de las herramientas más utilizadas ya que proporcionan información tanto cuantitativa sobre la producción científica como cualitativa, es decir sobre el impacto de esa producción

TORMES: an automated pipeline for whole bacterial genome analysis

El progreso de las tecnologías de secuenciación de alto rendimiento (HTS) y la reducción de los costes de secuenciación son tales que la secuenciación del genoma completo (WGS) ha sustituido a muchos ensayos y procedimientos tradicionales de laboratorio. La explotación del volumen de datos producidos por las plataformas de HTS requiere importantes conocimientos informáticos, lo que constituye el principal cuello de botella para la implantación de la WGS como técnica rutinaria de laboratorio. La forma de presentar la ingente cantidad de resultados a investigadores y clínicos sin conocimientos especializados en secuenciación genómica es también un problema importante. Nuestro trabajo como especialistas en técnicas de laboratorio tuvo que evolucionar hacia el aprendizaje bioinformático así que tras realizar el diploma en la universidad sevillana Pablo Olavide fuimos capaces de desarrollar el pipeline TORMES, en honor al río salmantino, un pipeline de fácil uso para el análisis WGS de bacterias de cualquier origen generadas por HTS en plataformas Illumina. Permite realizar un análisis genómico completo de un conjunto de bacterias (sin importar el número, especie u origen) partiendo directamente de los datos sin tratar procedentes de la plataforma de secuenciación. TORMES es de código libre y requiere seguir unas instrucciones sencillas para su instalación y uso, lo que lo convierte en una herramienta idónea para los usuarios sin conocimientos avanzados en bioinformática. TORMES está diseñado para usuarios no bioinformáticos, y automatiza los pasos necesarios para el análisis WGS directamente a partir de los datos de la secuencia en bruto: filtrado de la calidad de la secuencia, ensamblaje de novo, ordenación del genoma borrador frente a una referencia, anotación del genoma, tipado de secuencias multilocus (MLST), búsqueda de resistencia a antibióticos y genes de virulencia, y comparaciones de pangenomas. Una vez finalizado el análisis, TORMES genera un informe interactivo de tipo web que puede abrirse en cualquier navegador y ser compartido y revisado por los investigadores de forma sencilla. TORMES puede ejecutarse mediante comandos muy sencillos y representa una forma rápida y fácil de realizar análisis WGS