Magnetic resonance imaging of viral particle biodistribution in vivo.

We describe here a technique for the visualization of viral vector delivery by magnetic resonance imaging (MRI) in vivo. By conjugating avidin-coated baculoviral vectors (Baavi) with biotinylated ultra-small superparamagnetic iron oxide particles (USPIO), we are able to produce vector-related MRI contrast in the choroid plexus cells of rat brain in vivo over a period of 14 days. Ten microlitres of 2.5 x 10(10) PFU/ml nuclear-targeted LacZ-encoding Baavi with bUSPIO coating was injected into rat brain ventricles and visualized by MRI at 4.7 T. As baculoviruses exhibit restricted cell-type specificity in the rat brain, altered MRI contrast was detected in the choroid plexus of the injected ventricles. No specific signal loss was detected when wild-type baculoviruses or intact biotinylated USPIO particles were injected into the lateral ventricles. Cryosectioned brains were stained for nuclear-targeted beta-galactosidase gene expression, which was found to colocalize with MRI contrast. This study provides the first proof of principle for robust and non-invasive viral vector MRI by using avidin-displaying viruses in vivo. Considering the widespread use of MRI in current medical imaging, the approach is likely to provide numerous future applications in imaging of therapeutic gene transfer.

Photosynthetic and stomatal responses to high temperature and light in two oaks at the western limit of their range.

Bur oak (Quercus macrocarpa Michx.) and chinquapin oak (Q. muehlenbergii Engl.) leaves were exposed to high temperatures at various photosynthetic photon flux densities under laboratory conditions to determine if species-specific responses to these factors were consistent with the distribution of these oaks in gallery forests in the tallgrass prairies of northeastern Kansas, USA. Measurements of the ratio of chlorophyll fluorescence decrease, R(fd), indicated that chinquapin oak maintained greater photosynthetic capacity than bur oak across all tested combinations of irradiance (100, 400, 700 and 1000 micro mol m(-2) s(-1)) and temperature (40, 42, 44, 46 and 48 degrees C). In both oak species, manipulation of leaf temperature to about 47 degrees C for 45 min in the field led to a 45% decrease in carbon assimilation up to one week after the heat treatment, and to sharp reductions in stomatal conductance. Photosynthetic recovery patterns indicated that bur oak took longer to recover from heat stress than chinquapin oak, suggesting that heat stress may be important in determining distribution patterns of these oak species. Based on a comparison of the results with data from other forest species, we conclude that the photosynthetic temperature tolerances of bur oak and chinquapin oaks facilitate their dominance at the western limit of the eastern deciduous forest.