Production of ginkgolides and bilobalide by Ginkgo biloba plants and tissue cultures.

The accumulation of the terpenes ginkgolides and bilobalide in Ginkgo biloba was reported in plants as well as in plant cell cultures. Several hundred plants cultivated under controlled conditions in the field have been analyzed for their terpene production over many years. Cross-pollination experiments were performed with mature trees and the terpene content of the progeny was analyzed. The age of the tree is the main factor influencing the terpene content of the leaves as the level always decreases dramatically between young and old trees. 80 cell culture strains have been established and ginkgolides analyzed by GC/MS. These cell cultures reveal very low amounts of terpenes (1 microgram g-1 D.W. or less). On the contrary, isolated in vitro root cultures accumulate terpenes at the same concentration as the young plant leaves (4 mg g-1 D.W.). Attempts to obtain rapid growing roots or even hairy-roots did not succeed but the possibility to transform Ginkgo cell strains has been demonstrated.

Direct demonstration of the endocytic function of caveolae by a cell-free assay.

The endocytic function of caveolae was challenged by taking advantage of a cell-free assay directly measuring the detachment of receptor-containing vesicles from isolated plasma membranes. Plasma membranes from cultured cells surface-labeled with 125I-cholera toxin (segregating in caveolae) were isolated as described previously. Following incubation of these labeled membranes in the presence of nucleotide(s) and cytosol, a significant proportion of the initially membrane-associated radioactivity was released into the incubation medium in sedimentable form (14*10(6 )g). Results of biochemical, morphological, and fractionation analysis of the material containing the released radioactivity directly demonstrated that caveolae are plasma membrane domains involved in an endocytic process and resulting in the formation of caveolae-derived vesicles. In addition, these studies allowed a direct comparison of caveolae- and clathrin-coated pit-mediated endocytosis and reveal that these two processes diverge in terms of kinetics, cytosol and nucleotide requirements as well as in terms of the density and size of the endocytic vesicles formed.

Bi-directional gene switching with the tetracycline repressor and a novel tetracycline antagonist.

We have screened a panel of tetracycline (tc)-like compounds for their potential use with tc-repressor (tetR) based gene switches. The interaction between tc and tetR appears quite specific, as only tc itself and its close homologues anhydro-tc and doxycycline strongly inhibited DNA binding. However, a single tc-like compound, GR33076X, increased DNA binding of the tetR-VP16 fusion protein, both in eukaryotic cells and in bacteria. We provide evidence that this antagonist of tetracycline is potentially useful for accelerated gene switching, especially in whole animals.

Diagnosis of poststernotomy infection: comparison of three means of assessment.

Of 737 adults undergoing cardiac operations through a median sternotomy over a 2-year period (January 1988 to January 1990), sternal-substernal space infection requiring mediastinal exploration developed in 8. Six of these patients as well as 18 additional patients were evaluated for possible poststernotomy wound infection by computed tomography (4 true positive, 4 false positive, 10 true negative, 2 false negative), indium-111 leukocyte scanning (5 true positive, 0 false positive, 18 true negative, 1 false negative), and epicardial pacer wire cultures (6 true positive, 1 false positive, 12 true negative, 0 false negative). On the basis of this experience it is suggested that in addition to computed tomography, indium-111 leukocyte scanning and epicardial pacer wire cultures may be useful in the diagnosis of poststernotomy deep wound infection.

Isolation of Flavonol Glycosides from Ginkgo biloba Leaves.

6-Hydroxykynurenic acid and nine flavonol glycosides were isolated and identified from the leaves of GINKGO BILOBA: three of them were identified in Ginkgo for the first time, and two others are unusual P-coumaroyl esters.

Effects of luteinizing hormone (LH)-releasing hormone pulse amplitude and frequency on LH secretion by perifused rat anterior pituitary cells.

Recent studies have shown that LH secretion in vivo is pulsatile. In the present study, a cell perifusion system was employed to characterize the pituitary response to changes in LHRH pulse amplitude and frequency. Increases in pulse amplitude consistently elevated both mean LH levels and the amount of LH released in response to individual LHRH pulses. The EC50 for LHRH was approximately 3 nM. Increases in pulse frequency also increased mean LH levels, but frequencies of three or more pulses per h were associated with a decrease in the amount of LH released per pulse. Alterations in LHRH pulse characteristics changed qualitative as well as quantitative aspects of LH secretion, with high frequency, high amplitude pulses producing a biphasic response to LHRH. Initially a self-priming response was seen during the second and third hours of stimulation; this was followed by increasing desensitization of the cultures to LHRH. These results, by defining the pituitary response to specific conditions of stimulation, will help to clarify the relationship of LHRH stimulation to LH secretion in vivo.

Gonadal steroids modulate pulsatile luteinizing hormone secretion by perifused rat anterior pituitary cells.

Recent studies have shown that LH secretion is pulsatile and that LH pulse characteristics are affected by the prevailing steroid environment in both male and female rats. In the present study, a cell perifusion system was used to examine the effects of testosterone (T) and 17 beta-estradiol (E) on LHRH-stimulated pulsatile LH secretion. T inhibited LH secretion, increasing the EC50 for LHRH, while E stimulated secretion, lowering the EC50. Steroid effects were independent of both LHRH pulse amplitude and frequency. E also affected the pattern of LH secretion by facilitating both LHRH self-priming and desensitization to LHRH. These results show that steroids can affect pulsatile LH secretion by actions exerted at the pituitary level and that steroids can induce both quantitative and qualitative changes in LH secretion in the presence of an invariant LHRH stimulus. These results help to elucidate the mechanisms underlying steroid feedback in vivo, since reduction in pituitary responsiveness to LHRH may play an important role in T feedback, while facilitation by E of both self-priming and desensitization may serve to increase the magnitude and shorten the duration of the proestrous LH surge.

Mesenteric thrombosis following splenectomy.

Three cases of postsplenectomy mesenteric thrombosis, two associated with thrombocytosis, are presented. Experience has shown that persistent thrombocytosis, accompanied by abnormal platelet function, is not a benign condition and may be associated with thrombosis. When encountered, postsplenectomy thrombocytosis of greater than 800,000 per mm-3 must be evaluated by platelet function studies and anticoagulation begun. Post-prandial cramping abdominal pain may be an early symptom of thrombosis, demanding immediate anticoagulation. Low-dose heparin, ASA, and dipyridamole are three of the more commonly used treatment modalities. Small bowel resection is indicated if thrombosis occurs.