Estandarización de una prueba de inmunofluorescencia indirecta para IgG anti-Toxoplasma en ratón

We describe a technique based on indirect immunoflurescence antibody test (IFAT) for detection of specific anti-Toxoplasma IgG in mice. This technique detected infection from inocula with 10 tachyzoites of Toxoplasma in albino mice. In samples from human patients, this technique detected infection in amniotic fluid and in central nervous sample fluid. IFAT for IgG anti-Toxoplasma in mice is a useful complementary technique for the diagnosis of toxoplasmosis.Se describe una técnica de inmunofluorescencia indirecta (IFI) para la detección de IgG anti-Toxoplasma en ratón. La técnica reveló infecciones en el ratón a partir de inoculaciones de 10 taquizoitos de Toxoplasma gondii. En muestras de pacientes, se detectó infección en un líquido amniótico de una paciente con toxoplasmosis adquirida en el embarazo y en un líquido cefalorraquídeo de una paciente con toxoplasmosis cerebral. La técnica de inoculación en ratón y su detección por IFI es un complemento para el diagnóstico en el líquido amniótico o en el inmunosuprimido y es una herramienta valiosa para el aislamiento y caracterización de cepas de Toxoplasma

Mortality risk factors in primary Sjögren syndrome:a real-world, retrospective, cohort study

BACKGROUND: What baseline predictors would be involved in mortality in people with primary Sjögren syndrome (SjS) remains uncertain. This study aimed to investigate the baseline characteristics collected at the time of diagnosis of SjS associated with mortality and to identify mortality risk factors for all-cause death and deaths related to systemic SjS activity measured by the ESSDAI score.METHODS: In this international, real-world, retrospective, cohort study, we retrospectively collected data from 27 countries on mortality and causes of death from the Big Data Sjögren Registry. Inclusion criteria consisted of fulfilling 2002/2016 SjS classification criteria, and exclusion criteria included chronic HCV/HIV infections and associated systemic autoimmune diseases. A statistical approach based on a directed acyclic graph was used, with all-cause and Sjögren-related mortality as primary endpoints. The key determinants that defined the disease phenotype at diagnosis (glandular, systemic, and immunological) were analysed as independent variables.FINDINGS: Between January 1st, 2014 and December 31, 2023, data from 11,372 patients with primary SjS (93.5% women, 78.4% classified as White, mean age at diagnosis of 51.1 years) included in the Registry were analysed. 876 (7.7%) deaths were recorded after a mean follow-up of 8.6 years (SD 7.12). Univariate analysis of prognostic factors for all-cause death identified eight Sjögren-related variables (ocular and oral tests, salivary biopsy, ESSDAI, ANA, anti-Ro, anti-La, and cryoglobulins). The multivariate CPH model adjusted for these variables and the epidemiological features showed that DAS-ESSDAI (high vs no high: HR = 1.68; 95% CI, 1.27-2.22) and cryoglobulins (positive vs negative: HR = 1.72; 95% CI, 1.22-2.42) were independent predictors of all-cause death. Of the 640 deaths with available information detailing the specific cause of death, 14% were due to systemic SjS. Univariate analysis of prognostic factors for Sjögren-cause death identified five Sjögren-related variables (oral tests, clinESSDAI, DAS-ESSDAI, ANA, and cryoglobulins). The multivariate competing risks CPH model adjusted for these variables and the epidemiological features showed that oral tests (abnormal vs normal results: HR = 1.38; 95% CI, 1.01-1.87), DAS-ESSDAI (high vs no high: HR = 1.55; 95% CI, 1.22-1.96) and cryoglobulins (positive vs negative: HR = 1.52; 95% CI, 1.16-2) were independent predictors of SjS-related death.INTERPRETATION: The key mortality risk factors at the time of SjS diagnosis were positive cryoglobulins and a high systemic activity scored using the ESSDAI, conferring a 2-times increased risk of all-cause and SjS-related death. ESSDAI measurement and cryoglobulin testing should be considered mandatory when an individual is diagnosed with SjS.FUNDING: Novartis.</p

Libro: Las Ciencias Políticas y Sociales ante Contingencias de Amplio Impacto. Incógnitas y Propuestas

Ciencia Política, Administración Pública, Política y Gobierno, y Políticas Públicas. Licencia Creative Commons License 3.0 Reconocimiento-No Comercial-Sin Obras Derivadas. Usted es libre de copiar, distribuir y comunicar públicamente la obra bajo las condiciones siguientes: Reconocimiento - Debe reconocer los créditos de la obra de la manera especificada por el autor o el licenciador (pero no de una manera que sugiera que tiene su apoyo o apoyan el uso que hace de su obra). No comercial - No puede utilizar esta obra para fines comerciales. Sin obras derivadas - No se puede alterar, transformar o generar una obra derivada a partir de esta obra.Se analizan desde una perspectiva internacional a interdisciplinaria las vertientes, problemas, incógnitas y propuestas ante una nueva realidad o normalidad, resultado y consecuencia de la pandemia que se vive de manera contemporánea, de tal manera que la problematización abordada realimente propuestas, acciones y rutas adecuadas y satisfactorias que permitan la construcción de futuros promisorios.Academia Internacional de Ciencias Político-Administrativas y Estudios de Futuro, A.C. (IAPAS por sus siglas en inglés)

Retrospective evaluation of whole exome and genome mutation calls in 746 cancer samples

Funder: NCI U24CA211006Abstract: The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) curated consensus somatic mutation calls using whole exome sequencing (WES) and whole genome sequencing (WGS), respectively. Here, as part of the ICGC/TCGA Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium, which aggregated whole genome sequencing data from 2,658 cancers across 38 tumour types, we compare WES and WGS side-by-side from 746 TCGA samples, finding that ~80% of mutations overlap in covered exonic regions. We estimate that low variant allele fraction (VAF < 15%) and clonal heterogeneity contribute up to 68% of private WGS mutations and 71% of private WES mutations. We observe that ~30% of private WGS mutations trace to mutations identified by a single variant caller in WES consensus efforts. WGS captures both ~50% more variation in exonic regions and un-observed mutations in loci with variable GC-content. Together, our analysis highlights technological divergences between two reproducible somatic variant detection efforts

The importance of intrinsic traits, environment and human activities in modulating stress levels in a wild ungulate

Identifying the ecological and anthropogenic processes that affect wildlife physiology, and that may operate as chronic stressors, is of prime importance to implementing appropriate management and conservation strategies. Although advances have been made in understanding the physiological ecology of wild ungulates, little is known of how multiple intrinsic and ecological factors work, either independently or synergistically, to modulate their stress responses. By using faecal glucocorticoid metabolites (FGM) as indicators of stress, a set of environmental and human determinants affecting the stress physiology of wild red deer (Cervus elaphus) was examined in the Mediterranean ecosystems of south-western Europe, where this species is subjected to contrasting weather regimes and hunting management systems. Variation-partitioning techniques were also used to estimate the comparative influence of factors related to an individual's intrinsic characteristics, environmental conditions and management practices in shaping physiological stress levels. Our results showed that factors related to hunting management were the main drivers of FGM variation in red deer, followed by those related to the environmental conditions and individuals’ traits, and their effects were closely associated to spatio-temporal variability. Holding massive hunting events involving the use of hounds, as well as high population densities, were related to more long-term stress levels in the populations studied. Evidence was also found that supplementary feeding practices may mitigate the negative effects of reduced food availability in overabundant deer populations. Weather conditions were also significant factors explaining variation in stress levels; accumulated rainfall and an increase in ambient temperatures during the coldest months were associated with a decrease in stress hormone levels. No differences in hormonal concentrations were found between males and females, but higher levels of hormone metabolites were detected in younger animals in both sexes. Our findings provide an integrated perspective of how multiple factors impact on stress physiology in large wild herbivores and highlight the importance of considering management practices, as well as spatio-temporal variation, when assessing stress-inducing factors in wild populations. Given the implications of this study regarding the impact of human activities on physiological stress levels in wild animals, it could be an important basis to support wildlife management decisions.J.P.V. Santos was supported by a PhD Grant (SFRH/BD/65880/2009) from Fundação para a Ciência e a Tecnologia (FCT – Portugal) co-financed by the European Social Fund POPH-QREN programme. P. Acevedo is supported by the Spanish Ministerio de Economía y Competitividad (MINECO – Spain) and Universidad de Castilla-La Mancha (UCLM – Spain) through a ‘Ramón y Cajal’ contract (RYC-2012-11970). J. Carvalho and J. Queirós were both supported by PhD Grants (SFRH/BD/98387/2013 and SFRH/BD/73732/2010, respectively) from FCT.Peer Reviewe

Near infrared reflectance spectroscopy (NIRS) for predicting glucocorticoid metabolites in lyophilised and oven-dried faeces of red deer

Interest in measuring faecal glucocorticoid metabolites (FGM) as indicators of physiological homeostasis and performance in wildlife is increasing. However, current reference techniques, specifically enzyme immunoassays (EIAs) and radioimmunoassays (RIAs), are expensive, time-consuming, reagent-based, and the samples are destroyed during their application. Conversely, near infrared reflectance spectroscopy (NIRS) is a rapid, reagent-free and non-destructive technique, which, once calibrated by standard laboratory methods, can be used at a low cost. The objectives of this study were to evaluate the feasibility of using NIRS to predict glucocorticoid metabolite concentrations in red deer (Cervus elaphus) faeces, as well as the effect of lyophilisation and oven drying on FGM quantification. Seventy-eight fresh faecal samples were collected directly from the rectum of hunter-harvested red deer and then divided into two equal portions; one portion of each individual sample was lyophilised and the other portion was oven-dried. After dehydration, all faecal samples were ground and then analysed by RIA (standard laboratory technique) and scanned with an NIR spectrophotometer. Modified partial least squares regression was used to generate NIRS calibration equations for both lyophilised and oven-dried samples and a cross-validation procedure was employed for their optimisation. Near infrared reflectance spectroscopy proved to be a feasible, acceptably accurate and reliable technique for predicting FGM concentrations in red deer faeces subjected either to lyophilisation or to oven drying. Calibration and cross-validation results indicated that predictive equations for lyophilised faeces were slightly more precise and robust than for the oven-dried ones (lyophilised: R2 = 0.90, r2cv = 0.81, RPD = 2.72; oven-dried: R2 = 0.88, r2cv = 0.79, RPD = 2.26; CV: cross-validation, RPD: ratio of performance to deviation). Nevertheless, oven-dried faeces may be used as an alternative to lyophilised ones to quantify FGM levels accurately, provided that an appropriate combination of dehydration time and temperature is used during the desiccation process. High degrees of association and statistically significant positive correlations (p < 0.001) were found between the lyophilised and oven-dried samples regarding their FGM content, both for RIA assays and NIRS analyses. This study provides a new approach for assessing stress levels in free-ranging populations and has practical implications concerning wildlife monitoring as it makes it possible to improve the efficiency and reduce the cost and time constraints of current analytical techniques

Determining changes in the nutritional condition of red deer in Mediterranean ecosystems: Effects of environmental, management and demographic factors

Monitoring changes in the nutritional status of wild populations is crucial to understanding how species respond to natural or human-mediated variations in food availability and quality. It is a fundamental step for decision-making in wildlife management. Many studies on nutritional ecology still tend to rely on the use of single indicators. Nonetheless, nutrition has multiple interdependent facets that should be combined to better understand how organisms interact with their environment through feeding. Here, data from a large-scale cross-sectional survey conducted between 2010 and 2013 were used to examine the influence of environmental conditions and management practices on the variation of faecal nitrogen content (FN, index of dietary quality) and kidney fat index (KFI, indicator of nutritional condition) in red deer Cervus elaphus in the Mediterranean ecosystems of Iberia. Variations in the nutritional indicators in relation to sex, age, faecal fibre contents and faecal counts of lungworm larvae were also analysed. Our results showed that FN levels were statistically and positively correlated with vegetation primary productivity (P = 0.003). In contrast, faecal fibre fractions were significantly and negatively correlated with FN concentrations (P = 0.024 for hemicellulose, P < 0.0001 for cellulose and lignin). Moreover, a negative, marginally significant relationship was found between FN and the average altitude of the study areas (P = 0.055). The KFI was statistically higher in females than in males (P < 0.0001) and increased with increasing FN levels (P = 0.01). Fat stores were also statistically and positively correlated with vegetation productivity (P = 0.001) and with the amount of supplementary food provided (P = 0.04). The effect of high-quality diets on deer kidney fat deposits was gradually lower as the population density increased (P = 0.02). This result was more marked in non-food-supplemented populations. Both FN and KFI were statistically higher during the winter in relation to the autumn (P < 0.0001 and P = 0.005, respectively). This study shows that FN can be used to monitor and predict changes in the dietary quality, and the KFI in the nutritional status, in red deer in Mediterranean environments, reliably. Since both indicators showed different sensitivity to intrinsic, ecological and management determinants, this research also highlights their value and complementary use when studying populations of large wild herbivores.J.P.V. Santos was supported by a PhD Grant (SFRH/BD/65880/2009) from Fundação para a Ciência e a Tecnologia (FCT) co-financed by the European Social Fund POPH-QREN programme. J. Carvalho and J. Queirós were both also supported by PhD Grants (SFRH/BD/98387/2013 and SFRH/BD/73732/2010, respectively) from FCT. P. Acevedo is supported by Ministerio de Economía y Competitividad (MINECO) and University of Castilla-La Mancha through “Ramón y Cajal” contract (RYC-2012-11970). The present work also benefited from the financial aid of the research grant from MINECO (AGL2016-76358-R).Peer Reviewe