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Previous studies demonstrated that the Ras suppressor, RSU-1, localizes to human chromosome 10p13, a region frequently deleted in high grade gliomas, and that RSU-1 expression inhibited the tumorigenesis of a glioblastoma cell line. We have now examined RNA from human glial tumors for RSU-1 expression by RT-PCR using primers for the 5' and 3' ends of the RSU-1 open reading frame. Analysis of the amplified RSU-1 cDNA demonstrated that in addition to the entire 858 bp RSU-1 open reading frame, a shorter 725 bp RSU-1 fragment was amplified as well. Sequencing of this product revealed that it encoded a RSU-1 cDNA product which was missing a single 133 bp internal exon. This exon-deleted product was found in 30% of the high grade gliomas studied and 2/3 oligodendrogliomas, but not in other CNS tumors, bladder or colon tumors or normal tissue. The exon-deleted RSU-1 product was infrequently detected in RNA from human tumor cell lines. Expression of an HA-tagged form of the deleted RSU-1 protein in transfected Cos 1 cells revealed that the protein was unstable, with a half life of less than 1 h, in contrast to the full length HA-tagged Rsu-1 protein which was stable for more than 4 h. These results suggest that the alternative splicing of the RSU-1 RNA to produce the exon-deleted form constitutes a mechanism for reduction or loss of functional Rsu-1. Because the expression of Rsu-1 can inhibit malignant growth of glioblastoma cells, the depletion of Rsu-1, via the production of the alternatively spliced form of RSU-1, may inhibit growth regulation in tumors.

A natural shale and four synthetic organoclays were compared as potential sorbent additives to containment barriers at hazardous waste sites. Trimethylphenyl ammonium bentonite (TMPA-bent) was shown in batch experiments to have the greatest sorption capacities for 1,2,4-trichlorobenzene, trichloroethylene, and methyl isobutyl ketone, followed by the shale and a commercial organoclay. Sorption capacities were lowest for hexadecyltrimethyl ammonium bentonite (HDTMA-bent) and hexadecyl pyridinium bentonite (HDP-bent). Operative sorption mechanisms for the organoclays depended on the size of the organic modifier, i.e., uptake by the TMPA-bent occurred via adsorption onto mineral surfaces, while that for the HDTMA-bent and HDP-bent took place by absorption into organic phases formed by their long hydrocarbon tails. The shale was found to be by far the most cost-effective sorbent, an important factor for large scale applications. Solids concentration effects (i.e., higher apparent sorption capacities at lower experimental sorbent concentrations) were exhibited by HDTMA-bent and HDP-bent. This can be attributed to aggregation of sorbent particles as a result of interactions among their hydrocarbon chains. Solids effects were observed to decline and eventually disappear as sorbent concentrations were increased. Such effects must be considered in applying batch sorption results to flow-through systems.