Nucleic acid separations using superficially porous silica particles.

Ion pair reverse-phase liquid chromatography has been widely employed for nucleic acid separations. A wide range of alternative stationary phases have been utilised in conjunction with ion pair reverse-phase chromatography, including totally porous particles, non-porous particles, macroporous particles and monolithic stationary phases. In this study we have utilised superficially porous silica particles in conjunction with ion pair reverse-phase liquid chromatography for the analysis of nucleic acids. We have investigated a range of different pore-sizes and phases for the analysis of a diverse range of nucleic acids including oligonucleotides, oligoribonucleotides, phosphorothioate oligonucleotides and high molecular weight dsDNA and RNA. The pore size of the superficially porous silica particles was shown to significantly affect the resolution of the nucleic acids. Optimum separations of small oligonucleotides such as those generated in RNase mapping experiments were obtained with 80Å pore sizes and can readily be interfaced with mass spectrometry analysis. Improved resolution of larger oligonucleotides (>19mers) was observed with pore sizes of 150Å. The optimum resolution for larger dsDNA/RNA molecules was achieved using superficially porous silica particles with pore sizes of 400Å. Furthermore, we have utilised 150Å pore size solid-core particles to separate typical impurities of a fully phosphorothioated oligonucleotide, which are often generated in the synthesis of this important class of therapeutic oligonucleotide

Decline of Some West-Central Florida Anuran Populations in Response to Habitat Degradation

Recent reports have suggested that a global decline in amphibian populations has taken place during the past few decades. Urban development is thought to affect the richness and abundances of species and, therefore, could be an important cause of decline. We estimated the richness and abundances of anurans in wetlands at a residential development and in similar wetlands at a nearby undeveloped park. The residential development originally was pine flatwoods habitat, as is the undeveloped park curiently. We also compared the anuran species\u27 composition of the park in 1992 with the composition in 1974. Both richness and abundances of anurans in the residential development were different than those in the undeveloped park. Employing the same amount of sampling effort at both sites, we trapped or observed 11 species at the developement and 15 species at the park, and trapped 569 individuals at the development and 1224 individuals at the park. The anuran species richness at the undeveloped park in 1992 was nearly the same as in 1974; a single rare species apparently was not present in 1992. Of the 15 species present in both surveys, 14 showed higher abundances in 1992 than in 1974. We suggest that the current differences between the residential development and the park have resulted from degradation of both the uplands used by many species during the dry season and the temporary wetlands used by many species for reproduction. Four species especially sensitive to such degradation, Bufo quercicus, Scaphiopus h. holbrookii, Hyla femoralis, and H. gratiosa, were the species missing from the residential development. Not all species of anurans typical of pine flatwoods appeared to be affected adversely by development. Three species of ranids, Rana utricularia, R. grylio, and R. catesbeiana, were found in higher abundances at the residential development than at the park. These ranid species breed in a wide variety of aquatic systems, including the permanent bodies of water that are now abundant in the development, and probably use the uplands less than other anurans. If amphibian decline is international in scope, then the decline could be attributable either to global changes caused by humans, or to local, but widespread, environmental degradation, or to a combination of factors. While much recent popular focus has been on potential global causes of decline, we believe that this emphasis may have caused attention to be taken away from local causes that, as our study demonstrated, may be at least as important. We suggest that in many places, local environmental degradation is insidiously chipping away at amphibian diversity, and that more emphasis should be placed on these local causes than is now the case

Interactions Among Fire, Insects and Pathogens in Coniferous Forests of the Interior Western United States and Canada

1 Natural and recurring disturbances caused by fire, native forest insects and pathogens have interacted for millennia to create and maintain forests dominated by seral or pioneering species of conifers in the interior regions of the western United States and Canada. 2 Changes in fire suppression and other factors in the last century have altered the species composition and increased the density of trees in many western forests, leading to concomitant changes in how these three disturbance agents interact. 3 Two- and three-way interactions are reviewed that involve fire, insects and pathogens in these forests, including fire-induced pathogen infection and insect attack, the effects of tree mortality from insects and diseases on fuel accumulation, and efforts to model these interactions. 4 The emerging concern is highlighted regarding how the amount and distribution of bark beetle-caused tree mortality will be affected by large-scale restoration of these fire-adapted forest ecosystems via prescribed fire. 5 The effects of fire on soil insects and pathogens, and on biodiversity of ground-dwelling arthropods, are examined. 6 The effects of fire suppression on forest susceptibility to insects and pathogens, are discussed, as is the use of prescribed fire to control forest pests