A comparison of two in vitro maturation media for use with adult porcine oocytes for adult somatic cell nuclear transfer.

Two media used to mature adult porcine oocytes for somatic cell nuclear transfer were compared. In the first experiment, parthenogenetic embryos were produced using a maturation medium used by us previously to clone pigs (OMM199) and that described by Kühholzer et al. (2001) to transport oocytes overnight (BOMED). There was no difference in maturation rates between the two different media. However, BOMED medium increased the percentage of parthenogenetic embryos that developed to the blastocyst stage compared with OMM199 (49% vs. 29%, respectively). In a second experiment, BOMED medium increased the percentage of SCNT embryos that developed to the blastocyst stage compared with OMM199 (22% vs. 8%, respectively). The efficiency of our cloning protocol using adult oocytes matured in BOMED medium was then determined by transferring SCNT embryos reconstructed using adult fibroblasts to synchronized recipients. Primary cultures of adult fibroblasts were obtained from two adult male pigs and used for SCNT (passages 2-4). Between 82 and 146 fused couplets were transferred to seven recipients synchronized 1 day behind the embryos. Five recipients (71% pregnancy rate) subsequently farrowed a total of 23 piglets (4.4 average litter size). Overall efficiencies (liveborn/embryos transferred) were 3.2% for all transfers and 4.3% for animals that gave birth.

Nutrition and exercise environment available to outpatients, visitors, and staff in Children's hospitals in Canada and the United States.

BACKGROUND: Children's hospitals should advocate for children's health by modeling optimum health environments. OBJECTIVES: To determine whether children's hospitals provide optimum health environments and to identify associated factors. DESIGN: Telephone survey. SETTING: Canadian and US hospitals with accredited pediatric residency programs. PARTICIPANTS: Food services directors or administrative dietitians. MAIN OUTCOME MEASURES: Health environment grades as determined for 4 domains quantifying (1) the amount of less nutritious food sold at cafeterias (cafeteria grade), (2) the presence of fast food outlets (outlet grade), (3) the amount of nutritious food alternatives available (healthful alternative grade), and (4) the presence of patient obesity or employee exercise programs (program grade). RESULTS: The overall response rate was 87%. Compared with Canadian hospitals, US hospitals had more food outlets (89% vs 50%) and more snack/beverage vending machines (median, 16 vs 12) (P = .001 for both), despite equivalent consumer numbers. External companies managed more outlets at US vs Canadian hospitals (65% vs 14%; P = .01), and, generally, US hospitals recuperated more revenue from their outlets. Worst cafeteria grade was associated with US hospital location (odds ratio [OR], 8.9; 95% confidence interval [CI], 1.6-50; P = .01) and lower healthful alternative grade (OR, 0.016; 95% CI, 0.002-0.15; P<.001). Lower grade in any domain was related to whether hospitals received more revenue from noncafeteria food outlets (OR, 1.7; 95% CI, 1.06-2.72; P = .03) and the presence of more internally operated cafeterias (OR, 2.3 per cafeteria; 95% CI, 1.53-3.36; P<.001). CONCLUSIONS: Children's hospitals provide suboptimal health environments. Reliance on revenue may be an important motivating factor encouraging the adoption of outlets that serve less nutritious food.

An efficient method for producing alpha(1,3)-galactosyltransferase gene knockout pigs.

We have reported relatively efficient methods for somatic cell nuclear transfer and for knocking out the alpha(1,3)-galactosyltransferase (alpha1,3-GT) gene in porcine fetal fibroblasts using a nonisogenic promoterless construct approach. Here we report the production of alpha1,3-GT gene knockout pigs using these procedures. Seven alpha1,3-GT gene knockout cell clones were identified by long-range PCR from 108 neomycin resistant (neo(R)) colonies, giving a 6.5% targeting efficiency. Three cell clones were used for nuclear transfer. Nuclear transfer was performed using a fusion before activation protocol using in vitro-matured adult oocytes. Between 51 and 110 fused couplets were transferred to 10 recipients synchronized 1 day behind the embryos. Parturition was induced on day 115, and piglets were delivered by caesarean section. Four recipients gave birth to a total of 18 live piglets. All pigs were female, and all three clones resulted in the birth of live pigs. alpha1,3-GT gene knockout pigs were identified by long-range PCR and confirmed by Southern blot analysis. The efficiency (embryos transferred/piglets born) of our cloning protocol was 1.9% for all transfers and 4.6% for animals that gave birth.

The 'Golden Keys' to health - a healthy lifestyle intervention with randomized individual mentorship for overweight and obesity in adolescents.

OBJECTIVE: To conduct a pilot study designed to measure the impact of a healthy lifestyle intervention with or without individualized mentorship on adiposity, metabolic profile, nutrition and physical activity in overweight teens. METHODS: A total of 38 overweight adolescents (body mass index above the 85th percentile) 12 to 16 years of age, who were enrolled in a healthy lifestyle intervention program for six months, were randomly assigned to a nonmentored or individualized mentored intervention. RESULTS: For the entire cohort (final n=32), there was a nonstatistically significant reduction in mean (± SD) body mass index z score (2.08±0.38 to 2.01±0.47, P=0.07) and waist circumference (98±10 cm to 96±11 cm, P=0.08), and significant improvements in high-density lipoprotein level (1.08±0.24 mmol/L to 1.20±0.26 mmol/L, P<0.001), and low-density lipoprotein/high-density lipoprotein ratio (2.55±0.84 to 2.26±0.87, P<0.001) from baseline to the end of the intervention. Subjects consumed fewer high-calorie foods (3.9±1.9 to 3.0±1.5 servings/day, P=0.01) and snacks (9.7±5.5 to 6.8±4.0 servings/day, P=0.02), made fewer fast food restaurant visits (1.4±1.3 to 0.8±0.9 visits/week, P=0.02), and had less screen time (8.3±3.8 to 6.9±3.6 h/day, P=0.01). In addition, mentorship was found to be a feasible approach to supporting weight management in obese teens. Our study was underpowered to determine treatment effect, but promising modifications to lifestyle were observed despite the absence of statistically significant improvements in outcomes. CONCLUSIONS: The healthy lifestyle intervention improved subjects' lifestyles and lipid profiles, and the addition of mentorship in this context is feasible. A larger study with a longer intervention time is required to determine whether behavioural changes are associated with clinical improvement and to determine the role of mentorship in promoting lifestyle change.