Using multiplexed regulation of luciferase activity and GFP translocation to screen for FOXO modulators

<p>Abstract</p> <p>Background</p> <p>Independent luciferase reporter assays and fluorescent translocation assays have been successfully used in drug discovery for several molecular targets. We developed U2transLUC, an assay system in which luciferase and fluorescent read-outs can be multiplexed to provide a powerful cell-based high content screening method.</p> <p>Results</p> <p>The U2transLUC system is based on a stable cell line expressing a GFP-tagged FOXO transcription factor and a luciferase reporter gene under the control of human FOXO-responsive enhancers. The U2transLUC assay measures nuclear-cytoplasmic FOXO shuttling and FOXO-driven transcription, providing a means to analyze these two key features of FOXO regulation in the same experiment. We challenged the U2transLUC system with chemical probes with known biological activities and we were able to identify compounds with translocation and/or transactivation capacity.</p> <p>Conclusion</p> <p>Combining different biological read-outs in a single cell line offers significant advantages over conventional cell-based assays. The U2transLUC assay facilitates the maintenance and monitoring of homogeneous FOXO transcription factor expression and allows the reporter gene activity measured to be normalized with respect to cell viability. U2transLUC is suitable for high throughput screening and can identify small molecules that interfere with FOXO signaling at different levels.</p

Control del dolor postoperatorio en cirugía: relación entre cirugías, pauta analgésica, dosis y percepción del dolor

El dolor agudo postoperatorio (DAP) se produce como consecuencia de la estimulación nociceptiva sobre los distintos órganos y tejidos en una intervención quirúrgica. Se realizó un estudio prospectivo, de cohortes para valorar la intensidad de dolor percibido durante el ingreso hospitalario, en los pacientes intervenidos en Cirugía General, así como las variables que influyen en el mismo como sexo, edad, técnica quirúrgica, antecedentes quirúrgicos, carácter de la cirugía y pauta analgésica. Se recogió el dolor de los pacientes en las primeras 24 horas postoperatorias con una escala para la valoración de la percepción del dolor. Formaron parte del estudio 60 pacientes, quienes situaron la media del DAP, en 4,61 entre dolor moderado y dolor fuerte. Las diferentes intervenciones se agruparon en 4 categorías según características comunes: hernias, laparoscopias, laparotomías y eventroplastias y procedimientos de proctología. Se comparó la media del dolor con cada tipo de intervención y se observó un grado de significación de 0,007. Con los resultados del estudio se pudo concluir que: - El sexo influye en la percepción del DAP, siendo más alto en las mujeres. - La técnica quirúrgica influye en la percepción del dolor, siendo más dolorosas las cirugías de tipo laparotomías y eventroplastias, y las menos, las hernias. El resto de variables estudiadas no mostraros relación en la percepción del dolor.Grado en Medicin

Walking for functional life

Nas próximas décadas os indivíduos com mais de 60 anos duplicarão e os indivíduos com mais de 80 anos quadruplicarão. A perda de independência ocorre devido a vários fatores (restrições à mobilidade, fragilidade, etc.). Conclusões: Quando se prescreve exercicio ou programas de promoção da saúde tendo como alvo populações idênticas às deste estudo, deve existir uma atenção especial para as atividades de marcha e atividades que utilizam as mãos (motricidade fina), uma vez que estes componentes específicos parecem ter um maior impacto na aptidão física functional. Programas adequados e políticas adequadas mostraram contribuir para manter adultos mais velhos saudáveis e independentes ao longo do tempo com um impacto social, financial e suporte de saúde pública positivo. Os resultados apontam para a importância da prescrição de exercício conciso tendo como alvo adultos mais velhos. Deve ser dada mais atenção especificamente à marcha e a atividades manuais como componentes de um programa de exercício comunitário, como objetivo de aumentar o envelhecimento ativo e saudável.Também uma maior atenção deve ser dada às estruturas ambientais (parques, jardins públicos, etc.), de modo a aumentar as atividades de marcha para todos, especialmente para os mais velhos.info:eu-repo/semantics/publishedVersio

Effectiveness and costs of phototest in dementia and cognitive impairment screening

<p>Abstract</p> <p>Background</p> <p>To assess and compare the effectiveness and costs of Phototest, Mini Mental State Examination (MMSE), and Memory Impairment Screen (MIS) to screen for dementia (DEM) and cognitive impairment (CI).</p> <p>Methods</p> <p>A phase III study was conducted over one year in consecutive patients with suspicion of CI or DEM at four Primary Care (PC) centers. After undergoing all screening tests at the PC center, participants were extensively evaluated by researchers blinded to screening test results in a Cognitive-Behavioral Neurology Unit (CBNU). The gold standard diagnosis was established by consensus of expert neurologists. Effectiveness was assessed by the proportion of correct diagnoses (diagnostic accuracy [DA]) and by the kappa index of concordance between test results and gold standard diagnoses. Costs were based on public prices and hospital accounts.</p> <p>Results</p> <p>The study included 140 subjects (48 with DEM, 37 with CI without DEM, and 55 without CI). The MIS could not be applied to 23 illiterate subjects (16.4%). For DEM, the maximum effectiveness of the MMSE was obtained with different cutoff points as a function of educational level [k = 0.31 (95% Confidence interval [95%CI], 0.19-0.43), DA = 0.60 (95%CI, 0.52-0.68)], and that of the MIS with a cutoff of 3/4 [k = 0.63 (95%CI, 0.48-0.78), DA = 0.83 (95%CI, 0.80-0.92)]. Effectiveness of the Phototest [k = 0.71 (95%CI, 0.59-0.83), DA = 0.87 (95%CI, 0.80-0.92)] was similar to that of the MIS and higher than that of the MMSE. Costs were higher with MMSE (275.9 ± 193.3€ [mean ± sd euros]) than with Phototest (208.2 ± 196.8€) or MIS (201.3 ± 193.4€), whose costs did not significantly differ. For CI, the effectiveness did not significantly differ between MIS [k = 0.59 (95%CI, 0.45-0.74), DA = 0.79 (95%CI, 0.64-0.97)] and Phototest [k = 0.58 (95%CI, 0.45-0.74), DA = 0.78 (95%CI, 0.64-0.95)] and was lowest for the MMSE [k = 0.27 (95%CI, 0.09-0.45), DA = 0.69 (95%CI, 0.56-0.84)]. Costs were higher for MMSE (393.4 ± 121.8€) than for Phototest (287.0 ± 197.4€) or MIS (300.1 ± 165.6€), whose costs did not significantly differ.</p> <p>Conclusion</p> <p>MMSE is not an effective instrument in our setting. For both DEM and CI, the Phototest and MIS are more effective and less costly, with no difference between them. However, MIS could not be applied to the appreciable percentage of our population who were illiterate.</p

Diagnostic Accuracy, Effectiveness and Cost for Cognitive Impairment and Dementia Screening of Three Short Cognitive Tests Applicable to Illiterates

BACKGROUND: Illiteracy, a universal problem, limits the utilization of the most widely used short cognitive tests. Our objective was to assess and compare the effectiveness and cost for cognitive impairment (CI) and dementia (DEM) screening of three short cognitive tests applicable to illiterates. METHODS: Phase III diagnostic test evaluation study was performed during one year in four Primary Care centers, prospectively including individuals with suspicion of CI or DEM. All underwent the Eurotest, Memory Alteration Test (M@T), and Phototest, applied in a balanced manner. Clinical, functional, and cognitive studies were independently performed in a blinded fashion in a Cognitive Behavioral Neurology Unit, and the gold standard diagnosis was established by consensus of expert neurologists on the basis of these results. Effectiveness of tests was assessed as the proportion of correct diagnoses (diagnostic accuracy [DA]) and the kappa index of concordance (k) with respect to gold standard diagnoses. Costs were based on public prices at the time and hospital accounts. RESULTS: The study included 139 individuals: 47 with DEM, 36 with CI, and 56 without CI. No significant differences in effectiveness were found among the tests. For DEM screening: Eurotest (k = 0.71 [0.59-0.83], DA = 0.87 [0.80-0.92]), M@T (k = 0.72 [0.60-0.84], DA = 0.87 [0.80-0.92]), Phototest (k = 0.70 [0.57-0.82], DA = 0.86 [0.79-0.91]). For CI screening: Eurotest (k = 0.67 [0.55-0.79]; DA = 0.83 [0.76-0.89]), M@T (k = 0.52 [0.37-0.67]; DA = 0.80 [0.72-0.86]), Phototest (k = 0.59 [0.46-0.72]; DA = 0.79 [0.71-0.86]). There were no differences in the cost of DEM screening, but the cost of CI screening was significantly higher with M@T (330.7 ± 177.1 €, mean ± sd) than with Eurotest (294.1 ± 195.0 €) or Phototest (296.0 ± 196. 5 €). Application time was shorter with Phototest (2.8 ± 0.8 min) than with Eurotest (7.1 ± 1.8 min) or M@T (6.8 ± 2.2 min). CONCLUSIONS: Eurotest, M@T, and Phototest are equally effective. Eurotest and Phototest are both less expensive options but Phototest is the most efficient, requiring the shortest application time

Plan estratégico de la industria de la panela en el departamento de Piura

El presente plan estratégico tiene como finalidad proponer estrategias que permitan desarrollar la producción de la panela en Piura, de tal manera que, para el año 2017, este departamento sea reconocido por la capacidad de sus agricultores de producir panela con altos estándares de calidad, mediante la utilización de tecnologías que aumenten la productividad y contribuyan a mejorar la calidad de vida de la comunidad. Para llegar a este resultado se han planteado objetivos que sopesarán el crecimiento a corto y largo plazo, los cuales deberán ir acentuando un crecimiento sostenido de esta industria, la misma que todavía se desarrolla con procedimientos empíricos y sin mayor desarrollo tecnológico, pero que tiene un alto potencial de crecimiento, particularmente por la geografía, el clima, y la disponibilidad de recursos. Si bien es cierto que todavía no se puede competir por volumen, tal como lo hacen India y Colombia, la panela de Piura se está posicionando en mercados de mayor exigencia como un producto de alta calidad, por la integridad de sus componentes vitamínicos y minerales, otorgándole atributos diferenciadores que aumentan la calidad de este producto. Por otro lado, se han identificado estrategias que permitirán un crecimiento y desarrollo, tanto en volumen como en calidad, de esta industria, las mismas que se pretenden desarrollar mediante la colaboración de entidades nacionales y extranjeras, de tal manera que proporcionen un crecimiento integral de la industria, y que generen valor tanto a los productores como a la sociedad, por estar este producto inmerso en esta de manera tradicional y culturalmente, lo que permite diseñar un plan que posibilite el crecimiento económico y social de los productores de esta industria.This strategic plan has the purpose of to propose strategies that allow to propitiate the development of the panela in Piura's Department, in such a way that for the year 2017 Piura will be recognized by the capacity of his farmers to produce panela with standard high places of quality, by means of the utilization of technologies that raise the productivity and help to improve the quality of life of the community. To come to this result there have appeared aims that will have to weigh the growth of short and long term, which will have to go accentuating a growth supported of this industry, the same one that still develops with empirical procedures and without major technological development, but that possesses a potential high place of growth, particularly for the geography, the climate and availability of resources. Though it is true that still one cannot compete for volume, as India and Colombia do it, Piura's panela is positioning on markets of major exigency as a product of high quality, and that possesses the integrity of his vitamins and minerals, obtaining attributes that raise the quality of our product. On the other hand, there have been identified strategies that will allow a growth and development, so much in volume and quality this industry. The industry try to develop by means of the collaboration of national and foreign entities, in such a way that they provide an integral growth of the industry, and which generate value to producers and company, the same one that this immersed to this product of a traditional way and culturally, allowing us to develop a planning that allows the economic and social growth of the producers of the above mentioned industryTesi

Influence of the actions observed on cervical motion in patients with chronic neck pain: a pilot study

The aim of the present pilot study was to prove if the action-observation (AOb) improved the cervical range of motion (CROM) in patients with nonspecific chronic neck pain (CNP). Double blind pilot study. A total of 28 subjects were randomly assigned to an effective-movement group (n= 14) and an ineffective-movement group (n= 14). The follow-up consisted of: pretreatment, posttreatment and 10 min after second measurement (motor imagery). Outcome measures were CROM, and pressure pain detection thresholds (PPDTs). No statistical differences were found in baseline on CROM and on the PPDT. Test for independent groups revealed significant changes in cervical rotation movement. Both groups in posttreatment (P= 0.042; Cohen d= 0.81) and after 10 min (P= 0.019; Cohen d= 0.9). For intragroup PPDT, the Wilcoxon test revealed significant effects in the effective movement at C2 of the pre to 10-min post (P= 0.040). However, the ineffective movement revealed a significant reduction in PPDT in zygapophyseal joint of C5-C6 as the pre to post (P= 0.010) as the pre to 10-min post (P= 0.041) periods. In conclusions this pilot study demonstrated that the effective AOb produced significant changes versus ineffective AOb in the CROM and it could influences in PPT in subject with CNP immediately

Identification of oxidative stress related proteins as biomarkers for lung cancer and chronic obstructive pulmonary disease in bronchoalveolar lavage

Lung cancer (LC) and chronic obstructive pulmonary disease (COPD) commonly coexist in smokers, and the presence of COPD increases the risk of developing LC. Cigarette smoke causes oxidative stress and an inflammatory response in lung cells, which in turn may be involved in COPD and lung cancer development. The aim of this study was to identify differential proteomic profiles related to oxidative stress response that were potentially involved in these two pathological entities. Protein content was assessed in the bronchoalveolar lavage (BAL) of 60 patients classified in four groups: COPD, COPD and LC, LC, and control (neither COPD nor LC). Proteins were separated into spots by two dimensional polyacrylamide gel electrophoresis (2D-PAGE) and examined by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF/TOF). A total of 16 oxidative stress regulatory proteins were differentially expressed in BAL samples from LC and/or COPD patients as compared with the control group. A distinct proteomic reactive oxygen species (ROS) protein signature emerged that characterized lung cancer and COPD. In conclusion, our findings highlight the role of the oxidative stress response proteins in the pathogenic pathways of both diseases, and provide new candidate biomarkers and predictive tools for LC and COPD diagnosis. © 2013 by the authors; licensee MDPI, Basel, Switzerland.LPA is funded by Fondo de Investigación Sanitaria (PI081156) and Fondo de Investigación Sanitaria (PI1102688). MDP is funded by Fondo de Investigación Sanitaria (CD 09/00148). AN is funded by the Portuguese Ministry of Science, Technology and Superior Education—FCT (Fundação para a Ciência e para a Tecnologia: SFRH/BD/48341/2008). SMP is funded by Fondo de Investigación Sanitaria (CD 11/00153).Peer Reviewe

Quality of life of patients with metastatic pancreatic adenocarcinoma initiating first-line chemotherapy in routine practice

Background: Despite advances in surgery, radiotherapy, and chemotherapy, pancreatic adenocarcinoma often progresses rapidly and causes death. The physical decline of these patients is expected to impact their quality of life (QoL). Therefore, in addition to objective measures of effectiveness, the evaluation of health-related QoL should be considered a matter of major concern when assessing therapy outcomes. Methods: Observational, prospective, multicenter study including patients with metastatic pancreatic adenocarcinoma who started first-line chemotherapy in 12 Spanish centers. Treatment and clinical characteristics were recorded at baseline. Patients' health-related quality of life, ECOG, and Karnofsky index were measured at baseline, at Days 15 and 30, and every four weeks up to 6months of chemotherapy. Health-related quality of life was measured using the EORTC-QLQ-C30 and EQ-5D questionnaires. Other endpoints included overall survival and progression-free survival. Results: The study sample included 116 patients (median age of 65years). Mean (SD) scores for the QLQ-C30 global health status scale showed a significant increasing trend throughout the treatment (p=0.005). Patients with either a Karnofsky index of 70-80 or ECOG 2 showed greater improvement in the QLQ-C30 global health status score than the corresponding groups with better performance status (p 50 at baseline had significantly greater overall survival and progression-free survival (p=0.005 and p=0.021, respectively). No significant associations were observed regarding the EQ-5D score. Conclusions: Most metastatic pancreatic adenocarcinoma patients receiving first-line chemotherapy showed an increase in health-related quality of life scores throughout the treatment. Patients with lower performance status and health-related quality of life at baseline tended to greater improvement. The EORTC QLQ-C30 scale allowed us to measure the health-related quality of life of metastatic pancreatic adenocarcinoma patients receiving first-line chemotherapy

Adipokines, Biomarkers of Endothelial Activation, and Metabolic Syndrome in Patients with Ankylosing Spondylitis

Ankylosing spondylitis (AS) is a chronic inflammatory rheumatic disease associated with accelerated atherosclerosis and increased risk of cardiovascular (CV) disease. AS patients also display a high prevalence of features clustered under the name of metabolic syndrome (MeS). Anti-TNF- α therapy was found to be effective to treat AS patients by suppressing inflammation and also improving endothelial function. Previously, it was demonstrated that a short infusion of anti-TNF- α monoclonal antibodyinfliximab induced a rapid and dramatic reduction in serum insulin levels and insulin resistance along with a rapid improvement of insulin sensitivity in nondiabetic AS patients. The role of adipokines, MeS-related biomarkers and biomarkers of endothelial cell activation and inflammation seem to be relevant in different chronic inflammatory diseases. However, its implication in AS has not been fully established. Therefore, in this review we summarize the recent advances in the study of the involvement of these molecules in CV disease or MeS in AS. The assessment of adipokines and biomarkers of endothelial cell activation and MeS may be of potential relevance in the stratification of the CV risk of patients with AS