High-throughput screening of potential inhibitors for the metabolism of the investigational anti-cancer drug 5,6-dimethylxanthenone-4-acetic acid.

By screening potential inhibitors of drug metabolism using the in vitro models, potential drug-drug interactions in vivo may be predicted with the use of appropriate pharmacokinetic principles. This study aimed to develop a rapid screening system using human liver microsomes to efficiently identify the potential inhibitors of DMXAA metabolism. Initial IC50 was estimated by using a two-point method, and then Ki values were determined if required and compared with those initial IC50 values. More than 100 compounds including known substrates and inhibitors of human uridine diphosphate glucuronosyltransferases (UGTs) and cytochrome P450 (CYP), anti-cancer drugs and xanthenone analogues were screened for their inhibitory effect on DMXAA glucuronidation and 6-methylhydroxylation in human liver microsomes. Both metabolites of DMXAA, DMXAA acyl glucuronide (DMXAA-G) and 6-hydroxymethyl-5-methylxanthenone-4-acetic acid (6-OH-MXAA), formed in human liver microsomes were quantitated by validated HPLC methods. The results indicated that there was a significant relationship (r2 = 0.966, P < 0.001) between the two-point IC50 values and the apparent Ki values for 20 compounds showing significant inhibitory effects on DMXAA metabolism, suggesting the usefulness of the two-point determination for the initial screening of compounds. This study has been completed using a strategy for rapid HPLC analysis and thus provided early access to detailed information for potential inhibitors of DMXAA metabolism and allows for further DMXAA-drug interaction studies.

Food waste treatment in a community center.

For urban community composting centers, the proper selection and use of bulking agent is a key element in not only the cost but also the quality of the finished compost. Besides wood chips (WC) widely used as BA, readily usable cereal residue pellets (CRP) can provide biodegradable carbon and sufficient free air space (FAS) to produce stabilizing temperatures. The objective of the present project was to test at a community center, the effectiveness of CRP in composting food waste (FW). Two recipes were used (CRP with and without WC) to measure: FAS; temperature regimes, and; losses in mass, water, carbon and nitrogen. Both recipes were composted during three consecutive years using a 2 m(3) commercial in-vessel composter operated in downtown Montreal (Canada). For all recipes, FAS exceeded 30% for moisture content below 60%, despite yearly variations in FW and BA physical properties. When properly managed by the center operator, both FW and CRP compost mixtures with and without WC developed within 3 days thermophilic temperatures exceeding 50 °C. The loss of total mass, water, carbon and nitrogen was quite variable for both recipes, ranging from 36% to 54%, 42% to 55%, 48% to 65%, and 4% to 55%, respectively. The highest loss in dry mass, water and C was obtained with FW and CRP without WC aerated to maintain mesophilic rather than thermophilic conditions. Although variable, lower nitrogen losses were obtained with CRP and WC as BA, compared to CRP alone, as also observed during previous laboratory trials. Therefore and as BA, CRP can be used alone but nitrogen losses will be minimized by adding WC. Compost stabilization depends on operator vigilance in terms of aeration. The measured fresh compost density of 530-600 kg/m(3) indicates that the 2 m(3) in-vessel composter can treat 6.5 tons of FW/year if operated during 7 months.

Phosphorylation by Akt1 promotes cytoplasmic localization of Skp2 and impairs APCCdh1-mediated Skp2 destruction.

Deregulated Skp2 function promotes cell transformation, and this is consistent with observations of Skp2 overexpression in many human cancers. However, the mechanisms underlying elevated Skp2 expression are still unknown. Here we show that the serine/threonine protein kinase Akt1, but not Akt2, directly controls Skp2 stability by a mechanism that involves degradation by the APC-Cdh1 ubiquitin ligase complex. We show further that Akt1 phosphorylates Skp2 at Ser 72, which is required to disrupt the interaction between Cdh1 and Skp2. In addition, we show that Ser 72 is localized within a putative nuclear localization sequence and that phosphorylation of Ser 72 by Akt leads to cytoplasmic translocation of Skp2. This finding expands our knowledge of how specific signalling kinase cascades influence proteolysis governed by APC-Cdh1 complexes, and provides evidence that elevated Akt activity and cytoplasmic Skp2 expression may be causative for cancer progression.