Ex vivo analysis of topotecan: advancing the application of laboratory-based clinical therapeutics.

Topotecan is currently approved for relapsed small-cell lung cancer and ovarian cancer. Topotecan's efficacy in the second-line setting and novel mechanism of action suggest broad antitumour activity. We utilised a clinically validated, cell-death, ex vivo assay in human tumour explants to examine the activity profile of topotecan alone and in combination with other antitumour agents. Serial dilutions of topotecan alone and in combination with other cytotoxic agents were applied to biopsy specimens of non-small-cell lung cancer (NSCLC) and breast, colon, and prostate cancers. Dose-response curves were interpolated to provide 50% lethal concentrations (LC(50)). The degree of synergy (by median effect) and normalised Z-scores (raw scores converted to relative activity distributed around the mean) were then computed. Single-agent activity was observed for topotecan in all four tumour types. In 57 chemotherapy-naive specimens, NSCLC revealed the highest activity, demonstrated by the lowest LC(50) value (0.26+/-0.06 microg ml(-1); P=0.002). Overall, previously treated and chemotherapy-naive specimens revealed no significant differences in mean LC(50)'s. Synergy was observed for several combinations, including topotecan plus cisplatin in prostate and for topotecan plus 5-fluorouracil in breast cancers. The Z-score analyses conducted suggest activity for previously unexplored drug regimens, including topotecan plus 5-fluorouracil, vinorelbine, and mitomycin-C in NSCLC and breast cancer. Phase II studies are underway to determine the degree to which these ex vivo findings will translate into improved clinical results.

PAS kinase: an evolutionarily conserved PAS domain-regulated serine/threonine kinase.

PAS domains regulate the function of many intracellular signaling pathways in response to both extrinsic and intrinsic stimuli. PAS domain-regulated histidine kinases are common in prokaryotes and control a wide range of fundamental physiological processes. Similarly regulated kinases are rare in eukaryotes and are to date completely absent in mammals. PAS kinase (PASK) is an evolutionarily conserved gene product present in yeast, flies, and mammals. The amino acid sequence of PASK specifies two PAS domains followed by a canonical serine/threonine kinase domain, indicating that it might represent the first mammalian PAS-regulated protein kinase. We present evidence that the activity of PASK is regulated by two mechanisms. Autophosphorylation at two threonine residues located within the activation loop significantly increases catalytic activity. We further demonstrate that the N-terminal PAS domain is a cis regulator of PASK catalytic activity. When the PAS domain-containing region is removed, enzyme activity is significantly increased, and supplementation of the purified PAS-A domain in trans selectively inhibits PASK catalytic activity. These studies define a eukaryotic signaling pathway suitable for studies of PAS domains in a purified in vitro setting.

Use of xanthan gum in dietary management of diabetes mellitus.

Xanthan gum (12 g/day) was fed in muffins during either the first or second half of a 12-wk period of muffin feeding, to free-living subjects. Nine subjects were diabetic, having moderately elevated serum glucose but managing without insulin or hypoglycemic drugs, and four were nondiabetic controls. Before the study and at the end of the xanthan and xanthan-free periods, bloods were taken before and 2 h after an oral glucose load. The feeding of xanthan gum lowered fasting and postload serum glucose and reduced fasting levels of total plasma cholesterol in diabetic subjects. Xanthan gum also tended to lower fasting and postload levels of gastrin and gastric inhibitory polypeptide (GIP) and fasting levels of total and VLDL triglyceride and cholesterol in VLDL and LDL fractions. Subjects reported a sense of fullness after consuming xanthan muffins but no severe digestive symptoms.

Congenital malformations and variations in reproductive performance in the ferret: effects of maternal age, color and parity.

Demographic data of ferrets from a commercial breeding colony were analyzed for the effects of maternal age, parity and strain on reproductive performance and the frequency of gross congenital abnormalities observed at parturition. Litter size (mean +/- SEM) was found to be greatest for young, primiparous females (10.3 +/- 0.2) and decreased with advancing maternal age and parity to a cohort mean of 8.1 +/- 0.1 for third parity females 16 months of age. Age, parity or strain had no effect on 24-hour neonatal mortality (7%) or mortality from birth to weaning (20%) and an examination of the causes for death suggested that these rates can be reduced. The malformation rate from two cohorts of females whelping at different times of the year was low (less than 1.0%) and not significantly different. A higher frequency of malformed offspring was detected in females of low previous parity (0-2) than in those with three or more. Based on data obtained in this survey, the ferret would seem a valuable alternative, nonrodent species for teratologic investigations using currently recommended protocols.