The effect of l-thymidine, acyclic thymine and 8-bromoguanine on the stability of model G-quadruplex structures

Background Guanine-rich oligonucleotides are capable of forming tetrahelical structures known as G-quadruplexes with interesting biological properties. We have investigated the effects of site-specific substitution in the loops and in the tetrads model G-quadruplexes using thymine glycol nucleic acid (GNA) units, l-thymidine and 8-Br-2′-deoxyguanosine. Methods Modified oligonucleotides were chemically synthesized and spectroscopic techniques were used to determine the relative stability of the modified G-quadruplex. The double 8-BrdG-modified quadruplexes were further characterized by Nuclear Magnetic Resonance. Binding to thrombin of selected quadruplex was analyzed by gel electrophoresis retention assay. Results The most interesting results were found with a 8-bromoG substitution that had the larger stabilization of the quadruplex. NMR studies indicate a tight relationship between the loops and the tetrads to accommodate 8-bromoG modifications within the TBA. Conclusions The substitutions of loop positions with GNA T affect the TBA stability except for single modification in T7 position. Single l-thymidine substitutions produced destabilization of TBA. Larger changes on quadruplex stability are observed with the use of 8-bromoG finding a single substitution with the highest thermal stabilization found in thrombin binding aptamers modified at the guanine residues and having good affinity for thrombin. Double 8-BrdG modification in anti positions of different tetrads produce a conformational flip from syn to anti conformation of 8-Br-dG to favor loop-tetrad interaction and preserve the overall TBA stability

Exploring the interaction of Curaxin CBL0137 with G-Quadruplex DNA oligomers

Curaxins and especially the second-generation derivative curaxin CBL0137 have important antitumor activities in multiple cancers such as glioblastoma, melanoma and others. Although most of the authors suggest that their mechanism of action comes from the activation of p53 and inactivation of NF-kB by targeting FACT, there is evidence supporting the involvement of DNA binding in their antitumor activity. In this work, the DNA binding properties of curaxin CBL0137 with model quadruplex DNA oligomers were studied by 1H NMR, CD, fluorescence and molecular modeling. We provided molecular details of the interaction of curaxin with two G-quadruplex structures, the single repeat of human telomere d(TTAGGGT)4 and the c-myc promoter Pu22 sequence. We also performed 1H and 31P NMR experiments were also performed in order to investigate the interaction with duplex DNA models. Our data support the hypothesis that the interaction of curaxin with G-quadruplex may provide a novel insight into the DNA-binding properties of CBL0137, and it will be helpful for the design of novel selective DNA-targeting curaxin analogues

Baseline mitral regurgitation predicts outcome in patients referred for dobutamine stress echocardiography

Purpose: A number of parameters recorded during dobutamine stress echocardiography (DSE) are associated with worse outcome. However, the relative importance of baseline mitral regurgitation (MR) is unknown. The aim of this study was to assess the prevalence and associated implications of functional MR with long-term mortality in a large cohort of patients referred for DSE. Methods: 6745 patients (mean age 64.9±12.2 years) were studied. Demographic, baseline and peak DSE data were collected. All-cause mortality was retrospectively analyzed. DSE was successfully completed in all patients with no adverse outcomes. Results: MR was present in 1019 (15.1%) patients. During a mean follow up of 5.1±1.8 years, 1642 (24.3%) patients died and MR was significantly associated with increased all-cause mortality (p<0.001). With Kaplan-Meier analysis, survival was significantly worse for patients with moderate and severe MR (p<0.001). With multivariate Cox regression analysis, moderate and severe MR (HR 2.78; 95% CI 2.17 - 3.57; and HR 3.62; 95% CI 2.89 - 4.53, respectively) were independently associated with all-cause mortality. The addition of MR to C statistic models significantly improved discrimination. Conclusions: MR is associated with all-cause mortality and adds incremental prognostic information among patients referred for DSE. The presence of MR should be taken into account when evaluating the prognostic significance of DSE results

Endosseous dental implant fractures an analysis of 21 cases

Implant fracture is an infrequent cause of implant failure. The present study evaluates 21 fractured implants, with an analysis of patient age and sex, the type, length and diameter of the implant, positioning in the dental arch, the type of prosthetic rehabilitation involved, the number of abutments and pontics, the presence or absence of distal extensions or cantilevers, and loading time to fracture. Implant fracture was more common in males than in females (15:4), and the mean patient age was 56.9 years. Most cases (n = 19) corresponded to implant-supported fixed prostheses - 16 with cantilevers of different lengths- while only two fractured implants were supporting overdentures instead of fixed prostheses. The great majority of fractured implants (80.9%) were located in the molar and premolar regions, and most fractured within 3-4 years after loading. It is important to know and apply the measures required to prevent implant fracture, and to seek the best individualized solution for each case - though complete implant removal is usually the treatment of choic

Validación dimensional del Student Engagement Questionnaire (SEQ) en población universitaria española. Capacidades del alumno y entorno de enseñanza/aprendizaje

En este trabajo se validó el Study Engagement Questionnaire (SEQ) de Kember y Leung (2009) en una muestra española. Es un instru- mento diseñado para realizar una evaluación conjunta de los procesos de enseñanza-aprendizaje en la universidad y devolver feed-back a los profeso- res y a las instituciones para la mejora de estos procesos. Se utilizó una muestra de 805 sujetos de tres universidades valencianas. El cuestionario evalúa diversas capacidades del alumno así como la capacidad del profesor para diseñar un entorno de aprendizaje idóneo para aprender. Ha tenido varias versiones hasta conseguir una estabilidad notable en sus dimensiones (variables latentes) y en las relaciones entre las variables que contempla. Mediante análisis factorial confirmatorio se ha corroborado la estructura propuesta por los autores así como las relaciones entre las variables involu- cradas. El instrumento parece, pues, adecuado para ser utilizado en mues- tras españolas para el propósito para el que fue diseñado.In this work, the SEQ (Student Engagement Questionnaire - Kember & Leung, 2009) was validated with a Spanish sample. This instru- ment is designed to make a joint assessment of teaching-learning processes at university and to provide feedback to teachers and institutions to im- prove these processes. A sample of 805 subjects from three Valencian uni- versities was used. The questionnaire assesses the capabilities of several students, and the teacher’s ability to design an adequate learning environ- ment. Several versions have been developed until its dimensions (latent variables), and the relations between the involved variables, achieve high degree of stability. A confirmatory factor analysis corroborated the struc- ture proposed by the authors and the relations between the involved varia- bles. Hence the instrument seems suitable to be used in Spanish samples for the proposal it was designed for