Efficient expression of CFTR function with adeno-associated virus vectors that carry shortened CFTR genes

Adeno-associated virus (AAV)-based vectors have been shown to be effective in transferring the cystic fibrosis gene (CFTR) into airway epithelial cells in animal models and in patients. However, the level of CFTR gene expression has been low because the vector cannot accommodate the CFTR gene together with a promoter. In this study, we described a strategy to reduce the size of the CFTR cDNA to allow the incorporation of an effective promoter with the CFTR gene into AAV vectors. We engineered and tested 20 CFTR mini-genes containing deletions that were targeted to regions that may contain nonessential sequences. Functional analyses showed that four of the shortened CFTRs (one with combined deletions) retained the function and the characteristics of a wild-type CFTR, as measured by open probability, time voltage dependence, and regulation by cAMP. By using an AAV vector with a P5 promoter, we transduced these short forms of CFTR genes into target cells and demonstrated high levels of CFTR expression. We also demonstrated that smaller AAV/CFTR vectors with a P5 promoter expressed the CFTR gene more efficiently than larger vectors or a vector in which CFTR gene was expressed from the AAV inverted terminal repeat sequence. The CFTR mini-gene with combined deletions was packaged into AAV virions more efficiently, generated higher titers of transducing virions, and more effectively transferred CFTR function into target cells. These new vectors should circumvent the limitations of AAV vector for CFTR expression. Our strategy also may be applicable to other genes, the sizes of which exceed the packaging limit of an AAV vector.     

A primate model for enteral nutrition by tube

To establish an animal model for the controlled study of enteral nutrition by tube, five adult chair-adapted primates (Macaca fasicularis) had gastrostomy and jejunostomy tubes placed for the delivery of a modified protein isolate diet. Following 7 days of postoperative depletion with a hypocaloric infusion of dextrose (20 kcal, 0 g N/kg/day), the animals were repleted for 10 days with tube feedings (124 kcal, 0.73 g N/kg/day). There was no operative mortality or morbidity and each animal demonstrated conversion to anabolism by significant weight gain, positive nitrogen balance, and net protein synthesis as determined by [15N]glycine protein turnover rates. Significant correlation was found between caloric intake and nitrogen balance at the level of nitrogen provided in this diet (r = 0.88, p less than 0.05). This model was found to be well suited for the surgical and nutritional techniques required for the long-term study of enteral nutrition by tube.     

Examining cognitive function across the lifespan using a mobile application

Many studies conducted in a laboratory or university setting are limited by funding, personnel, space, and time constraints. In the present study, we introduce a method of data collection using a mobile application that circumvents these typical experiment administration issues. Using the application, we examined cross-sectional age differences in cognitive function. We obtained data from more than 15,000 participants and replicated specific patterns of age-related differences in cognition. Using a subset of these participants, we also examined the processing speed account of age-related cognitive differences, and the association of exercise and leisure activity with cognitive function across the lifespan. We discuss the relative advantages and disadvantages of data collection with a mobile application, and provide recommendations for the use of this method in research.     

A multiplicative model of software defect repair times

We hypothesize that software defect repair times can be characterized by the Laplace Transform of the Lognormal (LTLN) distribution. This hypothesis is rooted in the observation that software defect repair times are influenced by the multiplicative interplay of several factors, and the lognormal distribution is a natural choice to model rates of occurrence of such phenomenon. Conversion of the lognormal rate distribution to an occurrence time distribution yields the LTLN. We analyzed a total of more than 10,000 software defect repair times collected over nine products at Cisco Systems to confirm our LTLN hypothesis. Our results also demonstrate that the LTLN distribution provides a statistically better fit to the observed repair times than either of the two most widely used repair time distributions, namely, the lognormal and the exponential. Moreover, we show that the repair times of subsets of defects, partitioned according to the Orthogonal Defect Classification (ODC) scheme also follow the LTLN distribution. Finally, we describe how the insights that lead to the LTLN repair time model allow us to consider and evaluate alternative process improvement strategies.