A metabolite-profiling study of shock-frozen leaves of Isatis tinctoria L., an old indigo dye plant and medicinal herb, revealed a complex pattern of indigo-forming compounds with higher polarities than the known indigo precursors isatan B and indican. These highly unstable compounds underwent rapid post-harvest transformation and were not detected in air-dried leaves. The major indigo precursor, named isatan A (4), was isolated by rapid normal-phase and gel chromatography, along with isatan B (3). A full spectral data set of 3 showed that the previous structure assignment as 'indoxyl-5-ketogluconate' has to be revised to 1H-indol-3-yl beta-D-ribohex-3-ulopyranoside. Isatan A (4) was identified as 1H-indol-3-yl 6'-O-(carboxyacetyl)-beta-D-ribohex-3'-ulopyranoside. In aqueous solution, glycosides 3 and 4 occur as hydrates and undergo rapid hydrolysis under very mild acidic or basic conditions.

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Cold-pressed, non-raffinated evening primrose oil was found to contain lipophilic radical scavengers. A highly enriched fraction of these compounds could be obtained from the oil by extraction with aqueous ethanol and subsequent liquid-liquid partitioning with petroleum. LC-DAD-MS analysis revealed that the fraction contained three aromatic compounds with identical UV and ESI-MS spectra. The compounds were isolated by RP-HPLC and their structures established by chemical and spectroscopic means as 3-O-trans-caffeoyl derivatives of betulinic, morolic, and oleanolic acid. The morolic acid derivative was a new compound. The three esters exhibited pronounced radical scavenging activity against the stable 2,2-diphenyl-1-picrylhydrazyl radical and were potent inhibitors of neutrophil elastase and cyclooxygenase-1 and -2 in vitro. Commercial samples of evening primrose oils contained only traces of these lipophilic antioxidants.

Alterations in peripheral blood T-lymphocyte subsets have been implicated to play a role in the pathophysiology of estrogen deficiency-induced bone loss in postmenopausal women. The aim of this study was to investigate this hypothesis further by flow cytometric analysis of peripheral blood lymphocyte subpopulations together with bone histomorphometry using ovariectomized (OVX) rats as an experimental estrogen deficiency model. 104 female 6-month-old Wistar rats were either OVX or sham-operated (SHAM). Eight rats served as baseline controls. Groups of 8 SHAM and 8 OVX rats were killed 1, 2, 4, 6, 9, and 12 weeks after surgery. Blood was collected prior to sacrifice. In whole blood samples, subpopulations of peripheral lymphocytes were analyzed by flow cytometry using FITC- or PE-labeled monoclonal antibodies against rat CD5, CD4, and CD8. CD4 and CD8 positive T-lymphocytes were determined by a double-labeling technique. Serum samples were analyzed for estradiol and progesterone. The cancellous bone of the distal femoral metaphysis was analyzed by quantitative bone histomorphometry. Ovariectomy caused a statistically significant fall in serum estradiol and progesterone levels from 2 weeks postovariectomy until the end of the trial. Deterioration of cancellous bone structure and osteopenia in the distal femur of OVX rats became evident at 2 and 4 weeks postovariectomy, respectively. Flow cytometric analysis of peripheral blood lymphocytes revealed that, except for a transient increase in CD4 positive T-lymphocytes in OVX rats relative to SHAM animals at 1 week post-surgery, the number of CD5, CD4, or CD8 positive lymphocytes or the mean fluorescence per cell for these antigens in OVX rats remained unchanged throughout the study. Also the CD4+/CD8+ ratio of peripheral blood T-lymphocytes was not influenced by ovariectomy, and none of these parameters were significantly correlated with serum estradiol or progesterone levels. These results suggest that consistent changes in peripheral blood T-lymphocyte subpopulations are not demonstrable in OVX rats by the techniques applied in this study, and do not seem to play a pathogenetic role in the development of estrogen deficiency-induced bone loss in the rat.

Four groups of five fattening bulls each consumed a concentrate--wheat straw-diet (2.5:1) supplemented with either 0, 7, 14 or 21% ground rape seed for 350 days. Rape seed contained 427 g crude fat (ether extract) and 127 mg vitamin E per kg dry matter. The supplementation with rapeseed increased the fat concentrations in the rations from 25 to 50, 75 and 100 g, and of vitamin E from 11 to 19, 26 and 34 mg per kg dry matter. All bulls were slaughtered with about 560 kg body weight. Fatty acid composition of depot fat and of the fat of musc. long. dorsi were determined by gas liquid chromatography. Vitamin E concentrations in blood, depot fat and muscle were determined by HPLC. Oxidative stability of depot fat was measured as induction time by means of rancimat-test. Rape seed supplementation decreased C16-fatty acids and increased C18-fatty acids in depot and muscle fat. Muscle fat contained significantly more mono and poly unsaturated fatty acids (40.2 and 7.4%) than depot fat (33.5 and 2.0%, respectively). Rape seed supplementation enhanced significantly the vitamin E-concentrations in all body samples. In depot fat vit. E increased from 4.5 to 7.3, 8.5 and 14.9 micrograms/g. Induction time increased from 10.9 to 18.5, 16.1 and 19.5 h, when 0, 7, 14 or 21% rapeseed were added.