Efficient gene transfer to human endothelial cells using DNA complexed to adenovirus particles

Human endothelial cells have been found to be relatively refractory to various methods of DNA transfection currently in common use. By using a transfection method involving DNA complexed with replication-deficient adenovirus particles, we have shown that 20% of a population of cultured endothelial cells can be transfected and high levels of transient expression achieved. Both early-passage human umbilical vein endothelial cells and the continuous differentiated line of human endothelium-derived EA.hy926 cells are responsive to this method of transfection. Efficient DNA transfection of endothelial cells is important for studies of endothelium-specific promoters and is a potentially useful route for transgenic therapy.     

A note on the readability on the Position Analysis Questionnaire (PAQ).

The Position Analysis Questionnaire (PAQ) is a structured job analysis questionnaire composed of worker-oriented job elements. 4 readability indexes--the Date-Chall, the Flesch, the FOG (R. Gunning), and the SMOG (H. McLaughlin)--were applied to both the directions and questions of the PAQ. The PAQ directions reach a college readability level using the Flesch and SMOG indexes and reach the college graduate level using the Dale-Chall and FOG indexes. The PAQ questions reach the college graduate readability level on all 4 indexes. Implications regarding the use of the PAQ in light of its difficult level of readability are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Evidence of independent gene duplications during the evolution of archaeal and eukaryotic family B DNA polymerases

Eukaryotes and archaea both possess multiple genes coding for family B DNA polymerases. In animals and fungi, three family B DNA polymerases, alpha, delta, and epsilon, are responsible for replication of nuclear DNA. We used a PCR-based approach to amplify and sequence phylogenetically conserved regions of these three DNA polymerases from Giardia intestinalis and Trichomonas vaginalis, representatives of early-diverging eukaryotic lineages. Phylogenetic analysis of eukaryotic and archaeal paralogs suggests that the gene duplications that gave rise to the three replicative paralogs occurred before the divergence of the earliest eukaryotic lineages, and that all eukaryotes are likely to possess these paralogs. One eukaryotic paralog, epsilon, consistently branches within archaeal sequences to the exclusion of other eukaryotic paralogs, suggesting that an epsilon-like family B DNA polymerase was ancestral to both archaea and eukaryotes. Because crenarchaeote and euryarchaeote paralogs do not form monophyletic groups in phylogenetic analysis, it is possible that archaeal family B paralogs themselves evolved by a series of gene duplications independent of the gene duplications that gave rise to eukaryotic paralogs.     

Endothelial cell expression of testican mRNA

By screening random cDNAs from a continuous vascular endothelial cell line, EA.hy926, we identified a 5 kb mRNA that is expressed at high levels by this human cell line and by an early passage umbilical vein endothelial cell line. It is detected at lower levels in certain stromal cell lines, but it is not detected in most other cell lines tested, indicating that it represents a differentially expressed function rather than a ubiquitous or housekeeping function. This mRNA was readily detected in samples derived from most human organs as might be expected for a gene expressed in the vascular wall. Sequencing of the 5 kb mRNA reveals its identity with 3.5 kb of previously published testis-derived cDNA sequence called testican (Alliel et al., 1993). Differential expression of this gene by endothelial cells contributes a new perspective on the potential function of testican.