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First-generation adenovirus vectors (FG AdVs) expressing short-hairpin RNA (shRNA) effectively downregulate the expressions of target genes. However, this vector, in fact, expresses not only the transgene product, but also virus-associated RNAs (VA RNAs) that disturb cellular RNAi machinery. We have established a production method for VA-deleted AdVs lacking expression of VA RNAs. Here, we showed that the highest shRNA activity was obtained when the shRNA was inserted not at the popularly used E1 site, but at the E4 site. We then compared the activities of shRNAs against hepatitis C virus (HCV) expressed from VA-deleted AdVs or conventional AdVs. The VA-deleted AdVs inhibited HCV production much more efficiently. Therefore, VA-deleted AdVs were more effective than the currently used AdVs for shRNA downregulation, probably because of the lack of competition between VA RNAs and the shRNAs. These VA-deleted AdVs might enable more effective gene therapies for chronic hepatitis C. [ABSTRACT FROM AUTHOR]

Betalains are one of the major plant pigment groups found in some higher plants and higher fungi. They are not produced naturally in any plant species outside of the order Caryophyllales, nor are they produced by anthocyanin-accumulating Caryophyllales. Here, we attempted to reconstruct the betalain biosynthetic pathway as a self-contained system in an anthocyanin-producing plant species. The combined expressions of a tyrosinase gene from shiitake mushroom and a DOPA 4,5-dioxygenase gene from the four-o'clock plant resulted in successful betalain production in cultured cells of tobacco BY2 and Arabidopsis T87. Transgenic tobacco BY2 cells were bright yellow because of the accumulation of betaxanthins. LC-TOF-MS analyses showed that proline-betaxanthin (Pro-Bx) accumulated as the major betaxanthin in these transgenic BY2 cells. Transgenic Arabidopsis T87 cells also produced betaxanthins, but produced lower levels than transgenic BY2 cells. These results illustrate the success of a novel genetic engineering strategy for betalain biosynthesis. [ABSTRACT FROM AUTHOR]

Abstract Purpose  To quantitatively investigate the time course of changes in corneal irregular astigmatism and their relation to best spectacle-corrected visual acuity (BSCVA) in eyes with keratoconus. Methods  Subjects were 47 patients (57 eyes) with keratoconus who had undergone videokeratography examinations at least twice, with an interval of 1 year or longer between each examination. By means of Fourier series harmonic analysis, topography data were separated into spherical component, regular astigmatism, asymmetry, and higher order irregularity. Results  In eyes that showed apparent progression on color-coded maps, the yearly rate of change was significantly positive for spherical component (P = 0.03) and higher order irregularity (P = 0.006). In eyes with apparent progression, the yearly change rate of BSCVA was significantly positive (P = 0.001), but not in the eyes without apparent progression (P = 0.1655). In eyes with progression on maps, the yearly change rate of BSCVA was significantly correlated with the change rate of higher order irregularity per year (r = 0.770, P = 0.0004). Conclusions  The results indicated that progression of keratoconus is associated with deterioration of BSCVA, which is in turn significantly correlated with increases in corneal higher order irregularity. Jpn J Ophthalmol 2007;51:265–269 @ Japanese Ophthalmological Society 2007 [ABSTRACT FROM AUTHOR]