A requirement for Nedd9 in luminal progenitor cells prior to mammary tumorigenesis in MMTV-HER2/ErbB2 mice.

Overexpression of the NEDD9/HEF1/Cas-L scaffolding protein is frequent, and drives invasion and metastasis in breast, head and neck, colorectal, melanoma, lung and other types of cancer. We have examined the consequences of genetic ablation of Nedd9 in the MMTV-HER2/ERBB2/neu mouse mammary tumor model. Unexpectedly, we found that only a limited effect on metastasis in MMTV-neu;Nedd9(-/-) mice compared with MMTV-neu;Nedd9(+/+) mice, but instead a dramatic reduction in tumor incidence (18 versus 80%), and a significantly increased latency until tumor appearance. Orthotopic reinjection and tail-vein injection of cells arising from tumors, coupled with in vivo analysis, indicated tumors arising in MMTV-neu;Nedd9(-/-) mice had undergone mutational selection that overcame the initial requirement for Nedd9. To better understand the defects in early tumor growth, we compared mammary progenitor cell pools from MMTV-neu;Nedd9(-/-) versus MMTV-neu;Nedd9(+/+) mice. The MMTV-neu;Nedd9(-/-) genotype selectively reduced both the number and colony-forming potential of mammary luminal epithelial progenitor cells, while not affecting basal epithelial progenitors. MMTV-neu;Nedd9(-/-) mammospheres had striking defects in morphology and cell polarity. All of these defects were seen predominantly in the context of the HER2/neu oncogene, and were not associated with randomization of the plane of mitotic division, but rather with depressed expression the cell attachment protein FAK, accompanied by increased sensitivity to small molecule inhibitors of FAK and SRC. Surprisingly, in spite of these significant differences, only minimal changes were observed in the gene expression profile of Nedd9(-/-) mice, indicating critical Nedd9-dependent differences in cell growth properties were mediated via post-transcriptional regulation of cell signaling. Coupled with emerging data indicating a role for NEDD9 in progenitor cell populations during the morphogenesis of other tissues, these results indicate a functional requirement for NEDD9 in the growth of mammary cancer progenitor cells.

Water quality monitoring: the basis for watershed management in the Oldman River Basin, Canada.

The Oldman River flows 440 km from its headwaters in south-western Alberta, through mountains, foothills and plains into the South Saskatchewan River. Peak flows occur in May and June. Three major reservoirs, together with more than a dozen other structures, supply water to nine irrigation districts and other water users in the Oldman basin. Human activity in the basin includes forestry, recreation, oil and gas development, and agriculture, including a large number of confined livestock feeding operations. Based on the perception of basin residents that water quality was declining and of human health concern, the Oldman River Basin Water Quality Initiative was formed in 1997 to address the concerns. There was limited factual information, and at the time there was a desire for finger pointing. Results (1998-2002) show that mainstem water quality remains good whereas tributary water quality is more of a challenge. Key variables of concern are nutrients, bacteria and pesticides. Point source discharges are better understood and better regulated, whereas non-point source runoff requires more attention. Recent data on Cryptosporidium and Giardia species are providing benefit for focusing watershed management activities. The water quality data collected is providing a foundation to implement community-supported urban and rural better management practices to improve water quality.

Artifacts in trimethylsilyl derivatization reactions and ways to avoid them.

Trimethylsilyl derivatives are routinely employed in gas chromatography to increase the volatility and stability of organic compounds containing active hydrogens. Normally only the desired derivative is formed when organic compounds are derivatized with common silylation reagents. However, some compounds form additional unexpected derivatives or by-products (artifacts). Artifact formation leads to multiple peaks for the same compound or unexpected components in the gas chromatographic analysis of mixtures. This review includes silylation artifacts identified in our laboratory by mass spectrometry during the last 20 years and references to those found in the literature. Also, means of avoiding artifact formation are discussed in detail.