Optimización de un protocolo de extracción de ADN a partir de sangre bovina hemolizada y coagulada para la detección molecular de Anaplasma spp.

Anaplasma spp. bacteria cause anaplasmosis, a disease which negatively affects livestock production worldwide. Molecular detection by PCR requires efficient extraction of DNA from whole blood, which in turn depends on blood sample quality. Failures in sampling procedures and/or sample storage can lead to hemolysis and blood clotting, which can hamper diagnosis. An established DNA extraction protocol using Chelex® 100 resin was modified to optimize detection of Anaplasma spp. in hemolyzed and coagulated bovine blood samples, as well as reduce its cost. The optimized protocol extracted highly pure DNA effective in PCR analysis. Efficiency of the optimized protocol was compared with two commercial DNA extraction kits. When used in PCR detection of Anaplasma spp., the concordance values for all three were high (Cohen’s Kappa = 0.72). The optimized protocol is effective at extracting DNA from complex blood samples and is much less costly than commercial methods, a clear advantage when operating under limited budgets.La anaplasmosis es una enfermedad que puede afectar de manera significativa a la producción en el sector pecuario. Para la detección molecular por PCR de Anaplasma spp., se requiere de una extracción eficiente de ADN a partir de sangre completa, lo que a su vez depende de la calidad de la muestra de sangre. Fallas en los procedimientos de muestreo o conservación de las muestras pueden ocasionar hemolisis y coagulación de la sangre, lo que a su vez puede obstaculizar el diagnóstico.  El objetivo de esta investigación fue optimizar un protocolo casero de bajo costo que utiliza resina de Chelex® 100 y permite detectar de manera eficiente a Anaplasma spp. en 30 muestras de sangre bovina hemolizada y coagulada. La eficiencia del protocolo optimizado se comparó con dos kits comerciales de extracción de ADN mostrando una alta concordancia (Kappa de Cohen = 0.72) en la detección molecular de Anaplasma spp. por PCR. Se recomienda esta metodología para superar las limitaciones de investigación que pueden surgir al extraer ADN a partir de muestras de sangre complejas

Nuclear RNA purification by flow cytometry to study nuclear processes in plants

The nature of plant tissues has continuously hampered understanding of the spatio-temporal and subcellular distribution of RNA-guided processes. Here, we describe a universal protocol based on Arabidopsis to investigate subcellular RNA distribution from virtually any plant species using flow cytometry sorting. This protocol includes all necessary control steps to assess the quality of the nuclear RNA purification. Moreover, it can be easily applied to different plant developmental stages, tissues, cell cycle phases, experimental growth conditions, and specific cell type(s). For complete information on the use and execution of this protocol, please refer to and . The nature of plant tissues has continuously hampered understanding of the spatio-temporal and subcellular distribution of RNA-guided processes. Here, we describe a universal protocol based on Arabidopsis to investigate subcellular RNA distribution from virtually any plant species using flow cytometry sorting. This protocol includes all necessary control steps to assess the quality of the nuclear RNA purification. Moreover, it can be easily applied to different plant developmental stages, tissues, cell cycle phases, experimental growth conditions, and specific cell type(s)

Design method for estimation of unit value of extrication works after an accident using selected special technologies for removing the traffic obstacle on a motorway

Abstrakt Tato diplomová práce je dílčí součástí celkového projektu na stanovení všeobecné metodiky výpočtu nákladů na vyproštění nákladního automobilu. Zabývá se legislativou související s vyproštěním nákladních vozidel, analýzou nákladů a vyčíslení nákladů v důsledku odtahů u vybrané speciální techniky po nehodě. Práce je vodítkem pro znalce z ústavu soudního inženýrství, kteří na základě podkladů mohou snadněji stanovit postup a jednotkovou cenu vyprošťovacích prací po nehodě. Poté se zrychlí určování této ceny např. při soudních jednání a vyplacení zisků odtahovým firmám.Abstract (example) This thesis is a minor part of the overall project on the establishment of a general methodology of calculation of the cost of rescuing a lorry. It deals with legislation related to goods vehicle extrication, analysis of costs and costs as a result of exhausts for selected special equipment after an accident. The work is a guide for experts from the Institute of forensic engineering, on the basis of who can more easily establish a procedure and the unit price recovery work after the accident. Then speeds up determining the prices for example during the court proceedings and the payment of the profits of a business.

Epigenetic prediction of response to anti-PD-1 treatment in non-small-cell lung cancer: a multicenter, retrospective analysis

Background: Anti-programmed death-1 (PD-1) treatment for advanced non-small-cell lung cancer (NSCLC) has improved the survival of patients. However, a substantial percentage of patients do not respond to this treatment. We examined the use of DNA methylation profiles to determine the efficacy of anti-PD-1 treatment in patients recruited with current stage IV NSCLC. Methods: In this multicentre study, we recruited adult patients from 15 hospitals in France, Spain, and Italy who had histologically proven stage IV NSCLC and had been exposed to PD-1 blockade during the course of the disease. The study structure comprised a discovery cohort to assess the correlation between epigenetic features and clinical benefit with PD-1 blockade and two validation cohorts to assess the validity of our assumptions. We first established an epigenomic profile based on a microarray DNA methylation signature (EPIMMUNE) in a discovery set of tumour samples from patients treated with nivolumab or pembrolizumab. The EPIMMUNE signature was validated in an independent set of patients. A derived DNA methylation marker was validated by a single-methylation assay in a validation cohort of patients. The main study outcomes were progression-free survival and overall survival. We used the Kaplan-Meier method to estimate progression-free and overall survival, and calculated the differences between the groups with the log-rank test. We constructed a multivariate Cox model to identify the variables independently associated with progression-free and overall survival. Findings: Between June 23, 2014, and May 18, 2017, we obtained samples from 142 patients: 34 in the discovery cohort, 47 in the EPIMMUNE validation cohort, and 61 in the derived methylation marker cohort (the T-cell differentiation factor forkhead box P1 [FOXP1]). The EPIMMUNE signature in patients with stage IV NSCLC treated with anti-PD-1 agents was associated with improved progression-free survival (hazard ratio [HR] 0·010, 95% CI 3·29 × 10 −4–0·0282; p=0·0067) and overall survival (0·080, 0·017–0·373; p=0·0012). The EPIMMUNE-positive signature was not associated with PD-L1 expression, the presence of CD8+ cells, or mutational load. EPIMMUNE-negative tumours were enriched in tumour-associated macrophages and neutrophils, cancer-associated fibroblasts, and senescent endothelial cells. The EPIMMUNE-positive signature was associated with improved progression-free survival in the EPIMMUNE validation cohort (0·330, 0·149–0·727; p=0·0064). The unmethylated status of FOXP1 was associated with improved progression-free survival (0·415, 0·209–0·802; p=0·0063) and overall survival (0·409, 0·220–0·780; p=0·0094) in the FOXP1 validation cohort. The EPIMMUNE signature and unmethylated FOXP1 were not associated with clinical benefit in lung tumours that did not receive immunotherapy. Interpretation: Our study shows that the epigenetic milieu of NSCLC tumours indicates which patients are most likely to benefit from nivolumab or pembrolizumab treatments. The methylation status of FOXP1 could be associated with validated predictive biomarkers such as PD-L1 staining and mutational load to better select patients who will experience clinical benefit with PD-1 blockade, and its predictive value should be evaluated in prospective studies

Impact of COVID-19 on cardiovascular testing in the United States versus the rest of the world

Objectives: This study sought to quantify and compare the decline in volumes of cardiovascular procedures between the United States and non-US institutions during the early phase of the coronavirus disease-2019 (COVID-19) pandemic. Background: The COVID-19 pandemic has disrupted the care of many non-COVID-19 illnesses. Reductions in diagnostic cardiovascular testing around the world have led to concerns over the implications of reduced testing for cardiovascular disease (CVD) morbidity and mortality. Methods: Data were submitted to the INCAPS-COVID (International Atomic Energy Agency Non-Invasive Cardiology Protocols Study of COVID-19), a multinational registry comprising 909 institutions in 108 countries (including 155 facilities in 40 U.S. states), assessing the impact of the COVID-19 pandemic on volumes of diagnostic cardiovascular procedures. Data were obtained for April 2020 and compared with volumes of baseline procedures from March 2019. We compared laboratory characteristics, practices, and procedure volumes between U.S. and non-U.S. facilities and between U.S. geographic regions and identified factors associated with volume reduction in the United States. Results: Reductions in the volumes of procedures in the United States were similar to those in non-U.S. facilities (68% vs. 63%, respectively; p = 0.237), although U.S. facilities reported greater reductions in invasive coronary angiography (69% vs. 53%, respectively; p < 0.001). Significantly more U.S. facilities reported increased use of telehealth and patient screening measures than non-U.S. facilities, such as temperature checks, symptom screenings, and COVID-19 testing. Reductions in volumes of procedures differed between U.S. regions, with larger declines observed in the Northeast (76%) and Midwest (74%) than in the South (62%) and West (44%). Prevalence of COVID-19, staff redeployments, outpatient centers, and urban centers were associated with greater reductions in volume in U.S. facilities in a multivariable analysis. Conclusions: We observed marked reductions in U.S. cardiovascular testing in the early phase of the pandemic and significant variability between U.S. regions. The association between reductions of volumes and COVID-19 prevalence in the United States highlighted the need for proactive efforts to maintain access to cardiovascular testing in areas most affected by outbreaks of COVID-19 infection

The Andalusian eHealth Library, ten years of leadership

The Andalusian eHealth Library (BV-SSPA) was set up in 2006 under the scope of strategy number IV, Knowledge Management, of the II Quality Plan for the Andalusian Health System (2005-2008)1, which established the key process: To guarantee the knowledge exchange within the Health System, with the following specific aims: -To define and develop a scientific information strategy for the Health System. -To establish the Andalusian eHealth Library. -To coordinate the Andalusian Health documentary centres. -To establish strategic agreements with similar organizations. During its ten years of operation, the BV-SSPA has focused on providing Health System Professionals with the best scientific information available, in a timely manner, for clinical decision purposes, as well as research and training; in other words, the knowledge manager which integrates and coordinates the hospital libraries.Ye

Proceso de Enfermería en un paciente adulto mayor con infarto agudo de miocardio (IAM)

The professional practice of nursing has achieved a philosophy of responsibility and ethical behavior for the care of human life, sustenance that gives meaning to their practice, which contributes to the man obtaining well-being in all aspects during its life cycle . Nursing practice is aimed at providing care to the user according to their human needs, in any area where they are taking into account the biological, psychological, social and spiritual aspects. In addition, it is an important part of the health team, acting in collaboration with other disciplines. Its essence is the care of man in a holistic way, in all its dimensions, so it is necessary to apply a systematic method to assess, diagnose, plan, execute and evaluate their interventions in users in order to preserve and recover health . Nursing professionals require their own scientific foundations, crystallized in the nursing process, to guide their actions in a planned, systematized and organized way1. In particular, the nursing process performed in a patient with acute myocardial infarction (AMI), aims to identify real and potential health problems that allow decision making based on existing knowledge and evidence of nursing care to be applied In a systematic and organized way.El ejercicio profesional de la enfermería ha alcanzado una filosofía de responsabilidad y comportamiento ético para el cuidado de la vida humana, sustento que le da significado a su práctica, misma que contribuye para que el hombre obtenga bienestar en todos los aspectos durante su ciclo de vida. La práctica de enfermería esta dirigida a proporcionar cuidados al usuario de acuerdo a sus necesidades humanas, en cualquier ámbito donde se encuentre tomando en cuenta los aspectos biológico, psicológico, social y espiritual. Además, forma parte importante del equipo de salud, al actuar en colaboración con otras disciplinas. Su esencia es el cuidado del hombre de forma holística, en todas sus dimensiones, por lo que se requiere aplicar un método sistemático que permita valorar, diagnosticar, planear, ejecutar y evaluar sus intervenciones en los usuarios con el fin de conservar y recuperar la salud. Los profesionales de enfermería requieren de fundamentos científicos propios, cristalizados en el proceso de enfermería, que guíe en forma planeada, sistematizada y organizada sus acciones1. En particular, el proceso de enfermería realizado a un paciente con infarto agudo al miocardio (IAM), pretende identificar los problemas de salud reales y potenciales que permitan la toma de decisiones con base en el conocimiento y evidencia existente del cuidado de enfermería para ser aplicado en forma sistemática y organizada

Nuclear RNA purification by flow cytometry to study nuclear processes in plants

The nature of plant tissues has continuously hampered understanding of the spatio-temporal and subcellular distribution of RNA-guided processes. Here, we describe a universal protocol based on Arabidopsis to investigate subcellular RNA distribution from virtually any plant species using flow cytometry sorting. This protocol includes all necessary control steps to assess the quality of the nuclear RNA purification. Moreover, it can be easily applied to different plant developmental stages, tissues, cell cycle phases, experimental growth conditions, and specific cell type(s). For complete information on the use and execution of this protocol, please refer to and . The nature of plant tissues has continuously hampered understanding of the spatio-temporal and subcellular distribution of RNA-guided processes. Here, we describe a universal protocol based on Arabidopsis to investigate subcellular RNA distribution from virtually any plant species using flow cytometry sorting. This protocol includes all necessary control steps to assess the quality of the nuclear RNA purification. Moreover, it can be easily applied to different plant developmental stages, tissues, cell cycle phases, experimental growth conditions, and specific cell type(s)

Environmental Fate of RNA Interference Pesticides: Adsorption and Degradation of Double-Stranded RNA Molecules in Agricultural Soils

ISSN:0013-936XISSN:1520-585