CRISPR Activation in Mouse Trophoblast Stem Cells.

The placenta is a vital organ that regulates nutrient supply to the developing embryo during gestation. In mice, the placenta is composed of trophoblast lineage and mesodermal derivatives, which merge through the chorioallantoic fusion process in a critical event for the progression of placenta development. The trophoblast lineage is derived from self-renewing, multipotent cells known as mouse trophoblast stem cells (mTSCs). These cells are a valuable tool that allows scientists to comprehend the signals regulating major placental cell types' self-renewal and differentiation capacity. Recent advances in CRISPR-Cas9 genome editing applied in mTSCs have provided novel insights into the molecular networks involved in placentation. Here, we present a comprehensive CRISPR activation (CRISPRa) protocol based on the CRISPR/gRNA-directed synergistic activation mediator (SAM) method to overexpress specific target genes in mTSCs.

Spirometry and respiratory oscillometry: Feasibility and concordance in schoolchildren with asthma.

OBJECTIVE: The purpose of this study was to describe the feasibility of respiratory oscillometry (RO) in schoolchildren with asthma, and the concordance of its results with those of spirometry, to determine its clinical usefulness. METHODS: RO and spirometry were performed in 154 children (6 to 14-year-old) with asthma, following strict quality criteria for the tests. Their feasibility (probability of valid test, time of execution, number of maneuvers needed to achieve a valid test, and perceived difficulty) was compared. The factors that influence feasibility were analyzed with multivariate methods. FEV1, FEV1/FVC, FVC and FEF25-75 for spirometry, and R5, AX and R5-19 for RO, were converted into z-scores and their concordance was investigated through intraclass correlation coefficients (ICC) and kappa indices for normal/abnormal values. RESULTS: There were no differences in the probability of obtaining a valid RO or spirometry (83.1% vs. 81.8%, p = 0.868). RO required a lower number of maneuvers [mean (SD) 4.2 (1.8) versus 6.0 (1.6), p < 0.001] and less execution time [5.1 (2.7) versus 7.6 (2.4) minutes, p < 0.001], and patients considered it less difficult. Age increased the probability of obtaining valid RO and spirometry. The concordance of results between RO and spirometry was low, and only between zFEV1 and zAX could it be considered moderate (ICC = 0.412, kappa = 0.427). CONCLUSION: RO and spirometry are feasible in children with asthma. RO has some practical advantages, but the concordance of its results with spirometry is low.

Effect of infused norepinephrine dosage on pressure ulcers in perianesthesia care unit patients: a pilot study.

To determine whether treatment with moderate-high dose norepinephrine is a contributory cause increasing the risk of pressure ulcer development in critical postoperative, mechanically ventilated, hyperglycemic patients. A pilot clinical cohort study was conducted in a PACU in a tertiary hospital. A convenience sample of 16 hyperglycemic patients undergoing mechanical ventilation with at least 24 hours' stay in the PACU were divided into two groups of eight according to dosage of norepinephrine infusion at any moment during the days monitored. One group received ≥ 2.5 mg/hour, and the other received <2.5 mg/hour. The dose of norepinephrine administered by continuous venous infusion and the occurrence of stage II pressure ulcers were recorded. The proportion of patients with pressure ulcers in the group receiving moderate-high dose of norepinephrine was 50% compared with 0% in the low-dose group (P = .038). The odds ratio for daily average norepinephrine administered (mg/hour) was 2.21 (95% CI = 1.27 to +infinite); exact P = .0049. The odds ratio for daily average norepinephrine administered adjusting for weight (mg/kg/hour) was 3.05 (95% CI = 1.299 to +infinite); exact P = .0016. Moderate-high dose norepinephrine is a contributory cause that significantly increases the risk of pressure ulcer development in certain short-stay PACU patients.

[Childhood obesity: pediatricians' attitudes and opinions]. / Obesidad infantil: opiniones y actitudes de los pediatras.

OBJECTIVES: To identify the Spanish pediatricians' opinions and attitudes towards obesity, in relation with its treatment, prevention, and care organization, barriers they find in its treatment, and to know the tools they consider more useful to have for the management of obese children. METHODS: Mail survey posted to the primary care pediatricians of Castilla-León (Spain) exploring: willingness to act against obesity, utility of therapies and preventive strategies, barriers found in treating obese patients, most needed tools for treating obesity, autoefficacy, and support to an obese children management programme in primary care. RESULTS: There was a broad consensus in seeing obesity as an important health problem that demands action from pediatricians. This willingness to action decreased when bigger personal barriers were found. The most frequently encountered barriers came from the social milieu: easy access and advertisement of certain foods and beverages, lack of implication of parents, lack of perception of a weight problem in children and parents. Training was the most trusted tool. Pediatricians considered themselves as some or low efficacious in treating obesity. The effectiveness of an obese children management program in primary care was surpassed by the work it would need. CONCLUSIONS: In spite of their high willingness to act against obesity, pediatricians feel themselves limited because they find barriers mainly from the social and cultural milieus. They ask for training to fight pediatric obesity.