€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ *****ACTA PHARMACEUTICA HUNGARICA***** Bathori M Mathe I [Droplet countercurrent chromatography to isolate ecdysteroids from the herb Silene tatarica (L>)] Ellenaramu cseppmegoszlasos kromatorgafia alkalmazasa ekdiszteroidok izolalasara. In: Acta Pharm Hung (1996 May) 66(3):125-31 ISSN: 0001-6659 (Published in Hungarian) Our research project was to isolate ecdysteroids from Silene tatarica (L.) Pers., which plant belongs to Caryophyllaceae family. This plant as a species of the Silence genus has been considered as a good potential source of ecdysteroids, including new, hitherto never identified natural product(s). The raw material of isolation was the herba of Silene tatarica (L.) Pers. that was cultivated in the fields of the Ecological and Botanical Research Institute of the Hungarian Academy of Sciences (Vacratot, Hungary) and the plants were collected in June of 1994. The isolation procedure started with methanol extraction of the ground and milled herba. To improve the recovery, the extraction was done with boiling methanol under reflux for 20 minutes. The crude extract was purified using simple techniques, such as fractioned precipitation (with acetone), solvent-solvent partition (benzene-aqueous methanol). Further purification includes droplet counter-current chromatography (chloroform-methanol-water, 65:20:20, v/v/v) and straight-phase HPLC (silica stationary phase, dichloromethane-150 propanol-water. 125:25:2 v/v/v, mobile phase). Structural identification was done by using mass spectrometry and NMR spectroscopy. Three of the isolated ecdysteroids were identified as 20-hydroxyecdysone, 2-deoxy-20-hydroxyecdysone and 20-hydroxyecdysone- 22-benzoate these compounds have never been detected in Silene tatarica (L.) Pers. Structural elucidation of the fourt isolated ecdysteroid resulted in a new natural product with the chemical structure of 2-deoxy-20-hydroxyecdysone 22-benzoate. €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ *****BIOCHEMICAL GENETICS***** Kamsteeg J van Brederode J van Nigtevecht G Identification, properties, and genetic control of UDP-glucose: cyanidin-3-rhamnosyl-(1 leads to 6)-glucoside-5-O-glucosyltransferase isolated from petals of the red campion (Silene dioica). In: Biochem Genet (1978 Dec) 16(11-12):1059-71 ISSN: 0006-2928 An enzyme catalyzing the transfer of the glucosyl moiety of UDP- glucose to the 5-hydroxyl group of cyanidin-3-rhamnosyl-(1 leads to 6)-glucoside has been demonstrated in petal extracts of Silene dioica plants. This glucosyltransferase activity was not detectable in green parts of these plants. The enzyme activity is controlled by a single dominant gene M; no glucosyltransferase activity could be demonstrated in petals of m/m plants. The enzyme was purified eightyfold by PVP and Sephadex G50 chromatography. The glucosyltransferase had a pH optimum of 7.4, had a molecular weight of about 55,000, was stimulated by divalent metal ions, and had a "true Km" values of 0.5 x 10(-3) M for UDP-glucose and 3.6 x 10(-3) M for cyanidin-3-rhamnosylglucoside. Pelargonidin-3-rhamnosylglucoside also could serve as acceptor. The enzyme did not catalyze the glucosylation of the 5-hydroxyl group of cyanidin-3-glucoside, although in petals of M/- n/n mutants cyanidin-3,5-diglucoside is present. ADP-glucose could not serve as a glucosyl donor. €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ Kamsteeg J van Brederode J van Nigtevecht G Identification and properties of UDP-glucose: cyanidin-3-O- glucosyltransferase isolated from petals of the red campion (Silene dioica). In: Biochem Genet (1978 Dec) 16(11-12):1045-58 ISSN: 0006-2928 An enzyme catalyzing the transfer of the glucosyl moiety of UDP- glucose to the 3-hydroxyl group of cyanidin has been demonstrated in petal extracts of Silene dioica mutants with cyanidin-3-O-glucoside in the petals. This transferase activity was also present in young rosette leaves and calyces of these plants. The highest glucosyltransferase activity was found in petals of opening flowers of young plants. The enzyme was purified ninetyfold by PVP and Sephadex chromatography. The glucosyltransferase had a pH optimum of 7.5, had a "true Km value" of 4.1 x 10(-4) M for UDP-glucose and 0.4 x 10(-4) M for cyanidin chloride, and was not stimulated by divalent metal ions. Both p-chloromercuribenzoate and HgCl2 inhibited the enzyme activity. Pelargonidin chloride and delphinidin chloride at reduced rates also served as substrates. The enzyme did not catalyze the glucosylation of the 3-hydroxyl group of flavonols or the 5- hydroxyl group of anthocyanins. ADP-glucose could not serve as a glucosyl donor. The results of Sephadex G150 chromatography suggest that the glucosyltransferase can exist as dimer of about 125,000 daltons and as active monomers of 60,000 daltons. The genetic control of the glucosyltransferase activity is discussed. €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ *****FEBS LETTERS***** Vellekoop P Lugones L van Brederode J Purification of an UDP-glucose:flavone, 7-O-glucosyltransferase, from Silene latifolia using a specific interaction between the enzyme and phenyl-Sepharose. In: FEBS Lett (1993 Sep 6) 330(1):36-40 ISSN: 0014-5793 An UDP-glucose:flavonoid, 7-O-glucosyltransferase, from Silene latifolia was isolated from petals and purified 450-fold using a combination of gel-filtration, affinity chromatography and anion- exchange chromatography. Affinity chromatography on a phenyl- Sepharose CL-4B column in combination with elution with the substrate, isovitexin (6-C-glucosylapigenin), was an especially effective purification step. A purification factor between 10 and 20 could be reached using this column. A possible mechanism for the specific interaction of the enzyme with the phenyl-Sepharose will be discussed. This method of purification may also be applicable to other enzymes which use aromatic compounds as substrates. On a SDS- PAGE gel a band of 54 kDa, which co-purified with enzyme activity, could be detected in the purest fraction. Registry Numbers: EC 2.4.1.- (Glucosyltransferases) EC 2.4.1.81 (flavone 7-O-beta-glucosyltransferase) 71822-39-4 (phenyl-sepharose) 9012-36-6 (Sepharose) €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ *****GLYCOCONJUGATE JOURNAL***** Lhernould S Karamanos Y Lerouge P Morvan H Characterization of the peptide-N4-(N-acetylglucosaminyl) asparagine amidase (PNGase Se) from Silene alba cells. In: Glycoconj J (1995 Feb) 12(1):94-8 ISSN: 0282-0080 The peptide-N4-(N-acetylglucosaminyl) asparagine amidase (PNGase Se) earlier described [Lhernould S., Karamanos Y., Bourgerie S., Strecker G., Julien R., Morvan H. (1992) Glycoconjugate J 9:191-97] was partially purified from cultured Silene alba cells using affinity chromatography. The enzyme is active between pH 3.0 and 6.5, and is stable in the presence of moderate concentrations of several other protein unfolding chemicals, but is readily inactivated by SDS. Although the enzyme cleaves the carbohydrate from a variety of animal and plant glycopeptides, it does not hydrolyse the carbohydrate from most of the corresponding unfolded glycoproteins in otherwise comparable conditions. The substrate specificity of this plant PNGase supports the hypothesis that this enzyme could be at the origin of the production of 'unconjugated N-glycans' in a suspension medium of cultured Silene alba cells. Registry Numbers: EC 3.5. (Amidohydrolases) EC 3.5.1.52 (glycopeptidase) 7006-34-0 (Asparagine) 7783-20-2 (Ammonium Sulfate) €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ *****INTERNATIONAL UROLOGY AND NEPHROLOGY***** Grases F Melero G Costa-Bauza A Prieto R March JG Urolithiasis and phytotherapy. In: Int Urol Nephrol (1994) 26(5):507-11 ISSN: 0301-1623 The effects of seven plants with suspected application to prevent and treat stone kidney formation (Verbena officinalis, Lithospermum officinale, Taraxacum officinale, Equisetum arvense, Arctostaphylos uva-ursi, Arctium lappa and Silene saxifraga) have been studied using female Wistar rats. Variations of the main urolithiasis risk factors (citraturia, calciuria, phosphaturia, pH and diuresis) have been evaluated. It can be concluded that beneficial effects caused by these herb infusions on urolithiasis can be attributed to some disinfectant action, and tentatively to the presence of saponins. Specifically, some solvent action can be postulated with respect to uric stones or heterogeneous uric nucleus, due to the basifying capacity of some herb infusions. Nevertheless, for all the mentioned beneficial effects, more effective and equally innocuous substances are well known. €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€ *****PHYTOCHEMISTRY***** Lacaille-Dubois MA Hanquet B Cui ZH Lou ZC Wagner H Acylated triterpene saponins from Silene jenisseensis. In: Phytochemistry (1995 Sep) 40(2):509-14 ISSN: 0031-9422 From the roots of Silene jenisseensis a new trans-p-methoxycinnamoyl triterpene saponin has been isolated along with its cis-p- methoxycinnamoyl isomer as an inseparable mixture. Their structures were established by chemical means and spectroscopic methods including 1D- and 2D-homonuclear and heteronuclear correlation NMR spectroscopy as 3-O-[beta-D-galactopyranosyl-(1 --> 2)-beta-D- glucuronopyranosyl]-28-O-[beta-D-glucopyranosyl-(1 --> 2)-alpha-L- rhamnopyranosyl- (1 --> 2)-beta-D-4-O-trans-p-methoxycinnamoyl- fucopyranosyl] quillaic acid and its cis-isomer, respectively. They did not show any activity in the in vitro chemoluminescence granulocytes assay, but exhibited only a weak inhibitory effect in the cyclooxygenase inhibition assay. €€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€