Characterization of DNA-protein cross-links formed by treatment of L1210 cells and nuclei with bis(2-chloroethyl)methylamine (nitrogen mustard)

Proteins cross-linked to DNA after nitrogen mustard (HN2) treatment of cells or isolated nuclei were purified in CsCl gradients. The protein-DNA cross-links could be cleaved by incubation in dilute acid and could be stabilized by alkali pretreatment. These results indicate that proteins cross-linked to DNA by HN2 are bound to alkylated purines. Analysis of the DNA-bound proteins on NaDodSO4-polyacrylamide gels showed that primarily large nonhistone proteins are cross-linked to DNA in cells treated with HN2. Very little if any histone is cross-linked to the DNA. Comparison of DNA bound proteins from HN2-treated cells and HN2-treated nuclei showed that in general the same proteins are linked to DNA in both cases, but some qualitative and quantitative differences exist.     

Modeled Impacts of Cover Crops and Vegetative Barriers on Corn Stover Availability and Soil Quality

Environmentally benign, economically viable, and socially acceptable agronomic strategies are needed to launch a sustainable lignocellulosic biofuel industry. Our objective was to demonstrate a landscape planning process that can ensure adequate supplies of corn (Zea mays L.) stover feedstock while protecting and improving soil quality. The Landscape Environmental Assessment Framework (LEAF) was used to develop land use strategies that were then scaled up for five U.S. Corn Belt states (Nebraska, Iowa, Illinois, Indiana, and Minnesota) to illustrate the impact that could be achieved. Our results show an annual sustainable stover supply of 194 million Mg without exceeding soil erosion T values or depleting soil organic carbon [i.e., soil conditioning index (SCI)?>?0] when no-till, winter cover crop, and vegetative barriers were incorporated into the landscape. A second, more rigorous conservation target was set to enhance soil quality while sustainably harvesting stover. By requiring erosion to be <1/2 T and the SCI-organic matter (OM) subfactor to be >?0, the annual sustainable quantity of harvestable stover dropped to148 million Mg. Examining removal rates by state and soil resource showed that soil capability class and slope generally determined the effectiveness of the three conservation practices and the resulting sustainable harvest rate. This emphasizes that sustainable biomass harvest must be based on subfield management decisions to ensure soil resources are conserved or enhanced, while providing sufficient biomass feedstock to support the economic growth of bioenergy enterprises.     

Experiments on the origins of optical activity

Two recent reports claim that (1) aqueous L-aspartic acid polymerizes faster than D-Asplin the presence of kaolin at 90°, and (2) L-phenylalanine is adsorbed by kaolin more extensively than D-Phe at pH 6 (the reverse being true at pH 2). The novelty of these observations and their potential significance for the origin of optical activity has prompted us to duplicate these experiments using more sensitive methods. L- and D, L-Asp in 0.01M solution were incubated with kaolin at 90° for 8 days. Careful examination of the aqueous residues from such experiments failed to demonstrate any preferential polymerization of L-Asp over D-Asp, or indeed any significant gross polymerization of Asp at all. In other experiments 0.001M solutions of D, L-Phe at pH 6 and pH 2 were stirred with large excesses of kaolin for 24 hr, and the aqueous extracts from these mixtures were examined for gross adsorption using the amino acid analyzer. No significant gross adsorption was noted. We then looked for asymmetric adsorption in the aqueous residues using optical ratatory dispersion, gas chromatography and thin layer chromatography. By none of these analytical criteria could we find any evidence whatsoever for the preferential adsorption of D- versus L-Phe from either pH 6 or pH 2 solutions. Finally, in experiments bearing on the origin of optical activity by parity violation during β-decay, we have irradiated solid samples of D-, L- and D, L-leucine in a 61700 Ci Sr-90 source at Oak Ridge National Lab. for 1.34 yr (total dose: 4.2×10⁸ rad). Gas chromatographic examination of the (appropriately derivitized) recovered samples showed that the L-Leu was 16.7% decomposed, the D-Leu 11.4% and the D,L-Leu 13.8% decomposed. The recovered D,L-Leu sample had a gas-chromatographically determined enantiomeric composition of 50.8% D-leu and 49.2% L-Leu. These data, though very close to expermental error, may indicate a slight preferential radiolysis of L-Leu compared to D-Leu by the Bremsstrahlung from Sr-90 β-decay. These high intensity irradiation experiments are being continued on a prolonged basis in order to reach more definitive conclusions.