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Gen Comp Endocrinol. 2000 Oct;120(1):108-17.
High blood cortisol levels and low cortisol receptor affinity: is the chub, Leuciscus cephalus, a cortisol-resistant teleost?

Pottinger TG, Carrick TR, Appleby A, Yeomans WE.

Windermere Laboratory, NERC Centre for Ecology and Hydrology, The Ferry House, Ambleside, Cumbria, LA22 0LP, United Kingdom. tgeh.ac.uk

In contrast to the relatively minor intra- and interspecies differences in blood cortisol levels reported for salmonid species, there is a more pronounced distinction between cortisol levels among the Salmonidae and the Cyprinidae, with both basal and stress-induced cortisol levels markedly higher in the latter. This study shows that in the chub, Leuciscus cephalus, a widely distributed European cyprinid, mean blood cortisol levels during stress (1500 ng mL(-1)) exceeded those reported for most other species of fish and, even in unstressed chub, cortisol levels (50-100 ng mL(-1)) were within the range known to cause immunosuppression, growth retardation, and reproductive dysfunction in salmonid fish. The chub appears to be atypical only with respect to plasma cortisol levels; the levels of plasma glucose and plasma lactate in unstressed and stressed chub are similar to those reported for other species. Plasma levels of 11-ketotestosterone in males and 17beta-estradiol in females are lower than those reported for salmonids but similar to those reported for other cyprinid species and display clear stress-induced reduction. Comparative analysis of the binding characteristics of the trout and chub gill cortisol receptor revealed that the total number of binding sites in gill tissue for each species was similar (B(max); approximately 50-100 fmol mg(-1) protein). However, the affinity of the binding site for cortisol displayed an eightfold difference between the species (rainbow trout: K(d) approximately 6 nM; chub: K(d) approximately 50 nM). Therefore, the potentially adverse effects of high circulating levels of cortisol found both at rest and under conditions of stress in chub may be offset by the lower affinity of the cortisol receptor, rather than the abundance of target-tissue receptor sites. This strategy is similar to that reported for some glucocorticoid-resistant rodents and New World primates. 2000 Academic Press.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11042016&dopt=Abstract

Arch Toxicol. 2000 Sep;74(7):379-87.
Establishment of a novel in vitro system for studying the interaction of xenobiotic metabolism of liver and intestinal microflora.

Laube B, Winkler S, Ladstetter B, Scheller T, Schwarz LR.

GSF-National Research Centre for Environment and Health, Institute of Toxicology, Neuherberg/Munich, Germany.

We developed a new two-chamber system for the coculture of hepatocytes and fecal microflora under aerobic and anaerobic conditions, respectively, to investigate the sequential metabolism of chemicals by the liver and microflora in vitro. The culture device consisted of two chambers separated by a permeable polycarbonate membrane. In the aerobic compartment, hepatocytes were cultivated as a monolayer on the membrane and in the anaerobic compartment fecal microflora as a suspension. To characterize the metabolic capacity of the microflora and hepatocytes, various marker enzymes were studied. Azoreductase, nitroductase, beta-glucuronidase, beta-glucosidase and sulphatase were tested in the microflora of the feces from three volunteers who had had significantly different eating habits for years (daily meat, mixed diet, vegetarian). The microflora exhibited significant activities and the various enzymes differed only moderately in the samples from the three volunteers. For rat hepatocytes the activities of various cytochrome P450 forms and conjugating enzymes served as markers. The enzyme activities were tested in the coculture system during a 4-h culture period intended for the test protocol. Deethylation of ethoxycoumarin and 2alpha-, 6beta- and 16alpha-hydroxylation of testosterone decreased by about 30%, 25%, 40% and 20%, respectively, while there was no loss of glucuronidation and sulphonation of 3-OH-benzo(a)pyrene nor of glutathione conjugation of 1-chloro-2,4-dinitrobenzene during the 4-h culture period. The activities of the tested hepatic phase I and II enzymes were not changed after coculture of the hepatocytes with the microflora for 4 h. The applicability of the in vitro system for studying the metabolic interaction of liver and microflora was demonstrated using 7-ethoxycoumarin and the developmental drug EMD 57033, a thiadiazinon derivative from Merck KGaA, as model compounds. Both compounds were oxidized and conjugated by liver cells. In the coculture of hepatocytes and fecal microflora the resulting glucuronides and sulphoconjugates were split by hydrolytic enzymes of the intestinal microflora.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11043493&dopt=Abstract

Mol Endocrinol. 2000 Oct;14(10):1618-26.
Androgen receptor nuclear translocation is facilitated by the f-actin cross-linking protein filamin.

Ozanne DM, Brady ME, Cook S, Gaughan L, Neal DE, Robson CN.

Prostate Research Group, School of Surgical and Reproductive Sciences, Medical School, University of Newcastle upon Tyne, England.

The human androgen receptor (hAR) is a ligand-dependent transcription factor responsible for the development of the male phenotype. The mechanism whereby nuclear translocation of the hAR is induced by its natural ligand 5alpha-dihydrotestosterone is a phenomenon not fully understood. The two-hybrid interaction trap assay has been used to isolate proteins that interact with the hAR in an attempt to identify molecules involved in hAR transactivation and movement. We have identified the actin-binding protein filamin, a 280-kDa component of the cytoskeleton, as an hAR interacting protein. This interaction is ligand independent but is enhanced in its presence. The functional significance of this interaction was analyzed using a cell line deficient in filamin via transient expression of a green fluorescent protein-hAR chimera. In filamin-deficient cells this revealed that hAR remained cytoplasmic even after prolonged exposure to synthetic ligand. Nuclear shuttling was restored when this cell line regained wild-type expression of filamin. These data suggest a novel role for filamin, implicating it as an important molecule in AR movement from the cytoplasm to the nucleus.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11043577&dopt=Abstract

Drugs Aging. 2000 Sep;17(3):161-3.
Androgen receptor-blocking agents: potential role in pancreatic cancer.

Greenway BA.

Derriford Hospital, Plymouth, England.

The growth of pancreatic adenocarcinoma may be under the control of the sex steroid hormone testosterone, besides other unknown stimuli. This premise was based on the discovery of androgen receptors, together with the enzymes aromatase and 5alpha-reductase, which use testosterone as a substrate, in malignant tissue. The additional support of low circulating serum testosterone and its stimulatory role in an animal model using human tumour xenografts further enhance this suggestion. Confirmatory evidence has now come from a double-blind, placebo-controlled trial in patients with pancreatic cancer in which flutamide, the pure androgen receptor blocker, doubled survival duration over control patients. Flutamide was administered in a dosage of 250mg 3 times daily. Support for this finding has recently been presented at a meeting of the Pancreatic Society of Great Britain. It is now important for further confirmatory studies to be carried out and to use flutamide in combination with other current adjuvant therapies in patients with pancreatic cancer.

online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11043816&dopt=Abstract

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