Adeno-associated virus vector integration junctions

Vectors derived from adeno-associated virus (AAV) have the potential to stably transduce mammalian cells by integrating into host chromosomes. Despite active research on the use of AAV vectors for gene therapy, the structure of integrated vector proviruses has not previously been analyzed at the DNA sequence level. Studies on the integration of wild-type AAV have identified a common site-specific integration locus on human chromosome 19; however, most AAV vectors do not appear to integrate at this locus. To improve our understanding of AAV vector integration, we analyzed the DNA sequences of several integrated vector proviruses. HeLa cells were transduced with an AAV shuttle vector, and integrated proviruses containing flanking human DNA were recovered as bacterial plasmids for further analysis. We found that AAV vectors integrated as single-copy proviruses at random chromosomal locations and that the flanking HeLa DNA at integration sites was not homologous to AAV or the site-specific integration locus of wild-type AAV. Recombination junctions were scattered throughout the vector terminal repeats with no apparent site specificity. None of the integrated vectors were fully intact. Vector proviruses with nearly intact terminal repeats were excised and amplified after infection with wild-type AAV and adenovirus. Our results suggest that AAV vectors integrate by nonhomologous recombination after partial degradation of entering vector genomes. These findings have important implications for the mechanism of AAV vector integration and the use of these vectors in human gene therapy.     

Superfluid Drops: Dynamics of Pinch-Off and Sliding Motion

We present high resolution and high speed (5 µs) photographs of ⁴He drops undergoing pinch-off and sliding down a cesiated inclined plane. When a fluid drop is stretched and pulled apart by gravity, a balance of surface tension and inertia resugts in a striking icicle-shaped column of fluid which connects the two separating parts. The narrowest point of the icicle is an example of a finite-time singularity in the equations of motion. The tip radius of the icicle L obeys a power law L～τ ^2/3, where τ is the time before the moment of pinch-off. We have verified this for both superfluid and normal drops. Because of the boundary condition requiring zero velocity at a solid wall, sliding and rolling motion of drops on a substrate is a subtle issue even for conventional fluids. For example, calculations of the dissipation yields nonphysical infinities. We have analyzed video images of sliding superfluid drop motion and measured the acceleration of ⁴He droplets on a Cesium substrate.     

Computer Networks

Computer networks have the ability to bring the power of large machines to work on a single problem and to provide reliable computer services to large populations. They also may become an unmanageable structure that can cripple itself in a fashion akin to the great Northeast power failure in 1965. Imagine the following sequence: computer X does not have the sine subprogram but relies on computer Y for it; computer Y on the other hand solves the sine subprogram using the cosine subprogram which it doesn't have; computer Y therefore calls X for a cosine; X solves for cosine using sine which it asks Y for.& Of course, you say, no computer network would be so simplistic. But would you guarantee it could never happen for any set of computer resources among N computers-and that the network might not head for the buried recursive disaster like a lemming for a cliff?     

A 6.5 GHz-11.5 GHz source using a grid amplifier with a twistreflector

The authors have constructed and tested an oscillator using a grid amplifier with external feedback from a twist reflector. The twist reflector serves two functions; it changes the output polarization to match the input, and its position sets the feedback phase. This permits a wider tuning range than has been possible with previous grid oscillators. The source could be continuously tuned from 8.2 GHz to 11.0 GHz by moving the twist reflector. By moving the polarizer and mirror in the twist reflector independently, a 1.8-to-1 frequency range from 6.5 GHz to 11.5 GHz was achieved. The peak effective radiated power was 6.3 W at 9.9 GHz     

Seniority and criterion measures of job proficiency.

"On the basis of an analysis of 47 studies involving measures of the job proficiency of 2,462 workers employed in 39 different occupations, the average correlation between seniority and job proficiency is .17 with a standard error of correlation of .02. The type of measure of job success influences the relation between seniority and job success… [and]… the influence of seniority increases with the skill level of the occupations sampled… with skilled occupation the influence appears to be large relative to the magnitude of validity coefficients commonly obtained and should be taken into account and controlled whenever possible." (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Efficacy of timed embryo transfer with fresh and frozen in vitro produced embryos to increase pregnancy rates in heat-stressed dairy cattle

Our objective was to determine whether pregnancy rates in heat-stressed dairy cattle could be enhanced by timed embryo transfer of fresh (nonfrozen) or frozen-thawed in vitro-derived embryos compared to timed insemination. Ovulation in Holstein cows was synchronized by a GnRH injection followed 7 d later by PGF2 alpha and a second treatment with GnRH 48 h later. Control cows (n = 129) were inseminated 16 h (d 0) after the second GnRH injection. On d 7, a fresh (n = 133) or frozen-thawed (n = 142) in vitro-derived embryo was transferred to cows assigned for timed embryo transfer after categorizing the corpus luteum by palpation per rectum as 3 (excellent), 2 (good or fair), 1 (poor), and 0 (nonpalpable). Response to the synchronization treatment, determined by plasma progesterone concentration (ng/ml) < or = 1.5 on d 0 and > or = 2.0 on d 7, was 76.2%. Mean plasma progesterone concentration on d 7 increased as the quality of corpus luteum improved from category 0 to 3. Concentrations of progesterone in plasma were elevated (> or = 2.0 ng/ml) at 21 d in 64.7 (fresh embryo), 40.3 (frozen embryo), and 41.4 +/- 0.1% (timed insemination) of cows, respectively. Cows that received a fresh embryo had a greater pregnancy rate at 45 to 52 d than did cows that received a frozen-thawed embryo or timed insemination (14.3 > 4.8, 4.9 +/- 2.3%). Body condition (d 0) of cows influenced the pregnancy rate and plasma progesterone concentrations. In summary, timed embryo transfer with fresh in vitro-produced embryos in heat-stressed dairy cattle improved pregnancy rate relative to timed insemination.