Purification of the Neisseria meningitidis transferrin binding protein-2 (TBP2) to homogeneity using column chromatography

The interleukin-6 (IL-6) gene is expressed by various stimuli including cytokines or viral infections, such as human T-cell leukemia virus type I (HTLV-1). However, it has not been well established how HTLV-1 induces the expression of the IL-6 gene. In the present study, we demonstrated that HTLV-1-derived transactivator protein, p40tax, could stimulate endogenous IL-6 gene expression. Furthermore, we showed that the NF-kappa B binding site (IL-6 kappa B site) located between -74 and -62 upstream of the cap site of the IL-6 gene was an essential cis-acting element for p40tax-mediated transactivation of the IL-6 gene expression by utilizing a series of 5' deletion mutants of the IL-6 5' flanking region as well as a construct with a mutated IL-6 kappa B site. We identified the presence of two nuclear factor complexes that bound to the IL-6 kappa B site. One was constitutively expressed, and the other was inducible by p40tax. Taken together, HTLV-1 p40tax directly induces IL-6 gene expression through the IL-6 kappa B site, indicating the close association between IL-6 overproduction and HTLV-1 infection.     

A high efficiency technique for the generation of transgenic sugar beets from stomatal guard cells

An optimized protocol has been developed for the efficient and rapid genetic modification of sugar beet (Beta vulgaris L.). A polyethylene glycol-mediated DNA transformation technique could be applied to protoplast populations enriched specifically for a single totipotent cell type derived from stomatal guard cells, to achieve high transformation frequencies. Bialaphos resistance, conferred by the pat gene, produced a highly efficient selection system. The majority of plants were obtained within 8 to 9 weeks and were appropriate for plant breeding purposes. All were resistant to glufosinate-ammonium-based herbicides. Detailed genomic characterization has verified transgene integration, and progeny analysis showed Mendelian inheritance.     

Evidence for phosphorylation of CLN3 protein associated with Batten disease

While Aspergillus ficuum phytaseA (phyA) was rapidly inactivated by 1,2-cyclohexanedione and phenylglyoxal, both specific modifiers of arginine, phytaseB (phyB) showed a markedly different behavior. First, phyB was totally insensitive to 1,2-cyclohexanedione even in the presence of 0.2 M guanidinium hydrochloride; second, the enzyme showed a great deal of resistance to inactivation by phenylglyoxal. Taken together, these results indicate that the chemical environment of the active site of phyB is very different from that of the active site of phyA. Despite sequence similarities of the active site region in these two proteins, their differential behavior to arginine modifiers indicates that other parts of the protein play a role in the active site formation. We expected some differences in the structure since the proteins have dissimilar kinetic parameters and pH optima.     

Radiofluorinated L-m-tyrosines: new in-vivo probes for central dopamine biochemistry

PURPOSE: Recent information indicates that large, sustained wall shear stress gradients are a dominant hemodynamic parameter associated with the location and severity of atherosclerosis and myointimal hyperplasia. This study computes the spatial values of wall shear stresses and their gradients for three carotid artery bifurcation geometries. METHODS: A computational fluid dynamics program was used to solve the transient two-dimensional partial differential equations that describe fluid flow. Blood was treated as both a Newtonian and a non-Newtonian incompressible fluid. Solutions for the velocities, wall shear stresses, and wall shear-stress gradients were obtained for three carotid bifurcation geometries: a normal carotid bifurcation (similar to a primarily reconstructed carotid endarterectomy), a patch-reconstructed carotid endarterectomy, and a gradually tapered, low-angle carotid bifurcation (no carotid bulb). RESULTS: Computed velocity profiles closely match published experimental ones. Disturbed flow velocities are largest in the bulb segment of the normal carotid bifurcation. Peak and minimum wall shear stresses and peak shear stress gradients occurred in the lateral internal carotid artery wall. These were binodal in the normal or primarily reconstructed carotid artery, localized at the distal end of the patch-reconstructed carotid bifurcation, and minimal in the smooth, tapered carotid bifurcation. Wall shear stresses and their gradients were slightly higher for non-Newtonian than Newtonian fluids in the normal carotid artery but were similar in the other two geometric configurations. CONCLUSION: These results indicate that flow disturbances in general and wall shear stress gradients in particular are markedly reduced in carotid artery bifurcations that are smooth and gradually tapered and do not have a bulb. Abrupt geometric wall changes such as those occurring in the normal carotid bulb and at the distal end of a patch-reconstruction after carotid endarterectomy are harbingers of disturbed flow and high wall shear stress gradients. These results suggest that carotid endarterectomy reconstruction geometry characterized by a gradually tapered internal carotid artery may minimize the hemodynamically induced component of early myointimal hyperplasia and thrombosis and late atherosclerotic restenosis.     

Social interaction and sensorimotor gating abnormalities in mice lacking Dvl1

Mice completely deficient for Dvl1, one of three mouse homologs of the Drosophila segment polarity gene Dishevelled, were created by gene targeting. Dvl1-deficient mice are viable, fertile, and structurally normal. Surprisingly, these mice exhibited reduced social interaction, including differences in whisker trimming, deficits in nest-building, less huddling contact during home cage sleeping, and subordinate responses in a social dominance test. Sensorimotor gating was abnormal, as measured by deficits in prepulse inhibition of acoustic and tactile startle. Thus, Dvl1 mutants may provide a model for aspects of several human psychiatric disorders. These results are consistent with an interpretation that common genetic mechanisms underlie abnormal social behavior and sensorimotor gating deficits and implicate Dvl1 in processes underlying complex behaviors.