Illumina mate-paired DNA sequencing-library preparation using Cre-Lox recombination.

Standard Illumina mate-paired libraries are constructed from 3- to 5-kb DNA fragments by a blunt-end circularization. Sequencing reads that pass through the junction of the two joined ends of a 3-5-kb DNA fragment are not easy to identify and pose problems during mapping and de novo assembly. Longer read lengths increase the possibility that a read will cross the junction. To solve this problem, we developed a mate-paired protocol for use with Illumina sequencing technology that uses Cre-Lox recombination instead of blunt end circularization. In this method, a LoxP sequence is incorporated at the junction site. This sequence allows screening reads for junctions without using a reference genome. Junction reads can be trimmed or split at the junction. Moreover, the location of the LoxP sequence in the reads distinguishes mate-paired reads from spurious paired-end reads. We tested this new method by preparing and sequencing a mate-paired library with an insert size of 3 kb from Saccharomyces cerevisiae. We present an analysis of the library quality statistics and a new bio-informatics tool called DeLoxer that can be used to analyze an IlluminaCre-Lox mate-paired data set. We also demonstrate how the resulting data significantly improves a de novo assembly of the S. cerevisiae genome.

Efectos del entrenamiento físico de pacientes con fibrilación auricular en régimen de rehabilitación cardiovascular / Effects of physical training of patients with atrial fibrillation in cardiovascular rehabilitation regimen

Introducción: Aun son escasos los estudios que recomiendan los pacientes con fibrilación auricular en los programas de rehabilitación cardiaca. Objetivo: Evaluar los efectos del entrenamiento físico en pacientes con fibrilación auricular en régimen de rehabilitación cardiovascular. Metodología: Se realizó un estudio pre-experimental, prospectivo, tipo pretest- postest con 18 pacientes con fibrilación auricular que asistieron durante 12 semanas a las sesiones de rehabilitación cardiaca. Se evaluó el paciente al inicio y finaldel programa con ergometría con gases espirados, análisis de sangre y ecocardiograma transtorácico. Resultados: Predominaron el sexo masculino (72.2 %), el diagnóstico de miocardiopatías (33.3 %), la hipertensión arterial (100 %) y la fibrilación auricular persistente de larga duración (55.6 %). Se encontró una diferencia estadísticamente significativa para cada variable morfofuncional: índice de masa corporal (28.0±5.0 vs. 26.9±4.4, p< 0.0001), consumo de oxígeno pico (13.6±3.0 vs. 15.3±2.9, p<0.0001), consumo de oxígeno en el umbral anaerobio (9.9±2.2 vs. 10.7±2.0; p=0.007), equivalente ventilatorio para dióxido de carbono (27.7±4.7 vs. 26.6±4.7; p<0.0001), equivalente ventilatorio para el oxígeno (24.2±5.0 vs. 23.8±4.9 p=0.001), pulso de oxígeno (10.2±2.5 vs. 12.1±2.1; p< 0,0001), unidades metabólicas (3.8±0.9 vs. 4.3±0.9; p<0.0001) y fracción de eyección ventricular izquierda (55,7±8,7 vs. 58,2±7,5, p=0,003). Las variables metabólicas disminuyeron significativamente (p<0.0001). La clase funcional de Weber mejoró en 5 pacientes (27.8 %). Conclusiones: El entrenamiento físico de pacientes con fibrilación auricular durante 12 semanas de rehabilitación cardiovascular mejoró la capacidad funcional, con incrementos evolutivos de la fracción de eyección ventricular izquierda y disminución de las variables metabólicas, sin generar riesgos ni complicaciones.

Effects of physical training of patients with atrial fibrillation in cardiovascular rehabilitation regimen Introduction: There are still few studies that recommend patients with atrial fibrillation in cardiovascular rehabilitation programs. Objective: To evaluate the effects of physical training in patients with atrial fibrillation undergoing cardiovascular rehabilitation. Method: A pre-experimental, prospective, pretest-posttest study was carried out with 18 patients with atrial fibrillation who attended cardiac rehabilitation sessions for 12 weeks. The patient was evaluated at the beginning and end of the program with stress test with expired gases, blood test and transthoracic echocardiogram. Results: The male sex (72.2%), the diagnosis of cardiomyopathies (33.3%), arterial hypertension (100%) and longterm persistent atrial fibrillation (55.6%) predominated. A statistically significant difference was found for each morphofunctional variable: body mass index (28.0±5.0 vs. 26.9±4.4, p<0.0001), peak oxygen consumption (13.6±3.0 vs. 15.3±2.9, p<0.0001), oxygen at the anaerobic threshold (9.9±2.2 vs. 10.7±2.0; p=0.007), ventilatory equivalent for carbon dioxide (27.7±4.7 vs. 26.6±4.7; p<0.0001), ventilatory equivalent for oxygen (24.2± 5.0 vs. 23.8±4.9 p=0.001), oxygen pulse (10.2±2.5 vs. 12.1±2.1; p<0.0001), metabolic units (3.8±0.9 vs. 4.3±0.9; p<0.0001) and fraction of left ventricular ejection (55.7±8.7 vs. 58.2±7.5, p=0.003). The metabolic variables decreased significantly (p<0.0001). Weber's functional class improved in 5 patients (27.8%). Conclusions: Physical training of patients with atrial fibrillation during 12 weeks of cardiovascular rehabilitation improved functional capacity, with progressive increases in left ventricular ejection fraction and decrease in metabolic variables, without generating risks or complications.

Genome Sequence of Helicobacter heilmannii Sensu Stricto ASB1 Isolated from the Gastric Mucosa of a Kitten with Severe Gastritis.

Here we report the genome sequence of Helicobacter heilmannii sensu stricto ASB1 isolated from the gastric mucosa of a kitten with severe gastritis. Helicobacter heilmannii sensu stricto has also been associated with gastric disease in humans. Availability of this genome sequence will contribute to the identification of genes involved in the pathogen's virulence and carcinogenic properties.