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the crystal structure of progesterone 5beta-reductase from digitalis lanata defines a novel class of short chain dehydrogenases/reductases.progesterone 5beta-reductase (5beta-por) catalyzes the stereospecific reduction of progesterone to 5beta-pregnane-3,20-dione and is a key enzyme in the biosynthetic pathway of cardenolides in digitalis (foxglove) plants. sequence considerations suggested that 5beta-por is a member of the short chain dehydrogenase/reductase (sdr) family of proteins but at the same time revealed that the sequence motifs that in standard sdrs contain the catalytically important residues are missing. here we present ...200818032383
[studies on the efficacy of tetraploid forms of digitalis purpurea and digitalis lanata in guinea pigs]. 195413224339
[pharmacological properties of adicin, soviet alpha-acetyldigitoxin].adicin, soviet alpha-acetyldigitoxin, obtained from digitalis lanata ehrh., c. scrophulariaceae after isolation from it of celanid is a highly effective cardiotonic of the digitalis type of action. it exerts a favourable ino- and tonotropic and an adverse chronotropic action on the heart. this action is brought up in varied animal species and can be observed for a long time (over 3 hours). adicin produces no adverse effect on the coronary blood flow and it is conducive to the improvement of veno ...19807002595
investigating digitalis lanata for anabolic effect. 19685684381
comparative experimental studies on the antiinflammatory effect of the therapeutic agents "maraslavin" and "infusion of digitalis lanata 20:100", applied locally. 196314128109
[comparison of the effect of digitalis lanata standard drug with digoxin and cymarin on guinea pigs]. 19675632864
[could the digitalis lanata drug standard be replaced by a pure glycoside from digitalis lanata?]. 19675632863
[pharmacological evaluation of tetraploid drugs of digitalis purpurea and digitalis lanata in guinea pigs]. 195513254336
[chromatographic, chemical and biological research on extracts of digitalis lanata ehrh. cultivated in various environmental conditions]. 196314116471
molecular cloning and expression of progesterone 5beta-reductase (5beta-por) from isoplexis canariensis.a full-length cdna clone that encodes progesterone 5beta-reductase (5beta-por, ec 1.3.1.3) was isolated from isoplexis canariensis leaves. the reading frame of the ic5beta-por gene is 1170 nucleotides corresponding to 389 amino acids. the sphi /sali ic5beta-por fragment was cloned into the pqe vector system and then transformed into escherichia coli strain m15[prep4]. the gene was functionally expressed and the recombinant enzyme was characterised. k(m) and v(max) were calculated to be 0.215 mm ...200616858667
expression of recombinant digitalis lanata ehrh. cardenolide 16'-o-glucohydrolase in cucumis sativus l. hairy roots.the coding sequence for the digitalis lanata ehrh. cardenolide 16'-o-glucohydrolase was inserted downstream of the 35s promoter in the binary vector pbi121 resulting in plant expression vector pbi121cgh. cotyledon explants excised from 10-day-old seedlings of cucumis sativus l. were transformed using agrobacterium rhizogenes 15834 harbouring pbi121cgh. hairy roots were obtained from infected cotyledon explants in vitro 10 days after inoculation. pcr amplification of coding sequences for cgh i, r ...200616775721
the vep1 gene (at4g24220) encodes a short-chain dehydrogenase/reductase with 3-oxo-delta4,5-steroid 5beta-reductase activity in arabidopsis thaliana l.the arabidopsis thaliana vep1 gene product shows about 70% sequence identity to digitalis lanata progesterone 5beta-reductase, an enzyme considered to catalyze a key step in the biosynthesis of cardiac glycosides. a. thaliana does not accumulate cardenolides but protein extracts prepared from its leaves were capable of reducing progesterone to 5beta-pregnane-3,20-dione. a full-length cdna clone encoding a delta(4,5)-steroid 5beta-reductase (at5beta-str, ec 1.1.1.145/1.3.1.23), a member of the sh ...200919166903
it could have happened to van gogh: a case of fatal purple foxglove poisoning and review of the literature.although death owing to the toxic effects of the therapeutic digitalis lanata extract, digoxin has been reported, there are no reported cases of fatal digitalis purpurea (digitoxin) plant intoxication in humans in the literature. we describe a case of ingestion of digitalis purpurea in a 64-year-old man, which was fatal despite administration of digibind. a review of the literature and aspects of management of plant digitalis poisoning are discussed.200717968204
molecular cloning and heterologous expression of progesterone 5beta-reductase from digitalis lanata ehrh.a full-length cdna clone that encodes progesterone 5beta-reductase (5beta-por) was isolated from digitalis lanata leaves. the reading frame of the 5beta-por gene is 1170 nucleotides corresponding to 389 amino acids. for expression, a sph1/sal1 5beta-por fragment was cloned into the pqe vector and was transformed into escherichia coli strain m15[prep4]. the recombinant gene was functionally expressed and the recombinant enzyme was characterized. the k(m) and v(max) values for the putative natural ...200616386278
cardenolides in digitalis lanata cells transformed with ti-plasmids.crown galls were induced by transformation of leaves, leaf discs, and shoots of the plant digitalis lanata with the agrobacterium tumefaciens strains c58 pti c58, b6s3 pti b6s3, and a136 pti a6nctmr-338::tn5. integration of plasmid dna in the genome of d. lanata was demonstrated by hybridization experiments. the transformed cells synthesized opines and showed hormone-autotrophic growth. the crown galls formed on leaves of d. lanata plants contained digitoxigenin derivatives (up to 0.8 muimol dig ...199017221446
experimental data about anabolic effect of digitalis lanata. 19685687106
progesterone 5ß-reductase of erysimumcrepidifolium: cdna cloning, expression in escherichia coli, and reduction of enones with the recombinant protein.erysimum is a genus of the brassicaceae family closely related to the genus arabidopsis. several erysimum species accumulate 5ß-cardenolides. progesterone 5ß-reductases (p5ßrs) first described in digitalis species are thought to be involved in 5ß-cardenolide biosynthesis. p5ßrs belong to the dehydrogenase/reductase super-family of proteins. a full length cdna clone encoding a p5ßr was isolated from erysimumcrepidifolium leaves by 5'/3' race-pcr (termed ecp5ßr). subsequently, the p5ßr cdnas of an ...201121767854
systemic induction of the biosynthesis of terpenic compounds in digitalis lanata.a bacterial screening was carried out in the rhizosphere of two digitalis species, d. thapsi and d. parviflora, both at the vegetative stage and at flowering. a total of 480 isolates were characterised at genus level, bacillus being the dominant genera in all cases. fifty percent of the bacillus strains isolated from each species were analysed by pcr-rapds. at 85% similarity, 12 groups separated for d. thapsi and 18 for d. parviflora. one strain of each group was selected for biological assay on ...200312685026
functional replacement of the essential ess1 in yeast by the plant parvulin dlpar13.a functionally pin1-like peptidyl-prolyl cis/trans isomerase (ppiase(1)) was isolated from proembryogenic masses (pems) of digitalis lanata according to its enzymatic activity. partial sequence analysis of the purified enzyme (dlpar13) revealed sequence homology to members of the parvulin family of ppiases. similar to human pin1 and yeast ess1, it exhibits catalytic activity toward substrates containing (thr(p)/ser(p))-pro peptide bonds and comparable inhibition kinetics with juglone. unlike pin ...200111118437
cloning and functional expression in escherichia coli of a cdna encoding cardenolide 16'-o-glucohydrolase from digitalis lanata ehrh.a clone of cardenolide 16'-o-glucohydrolase cdna (cgh i) was obtained from digitalis lanata which encodes a protein of 642 amino acids (calculated molecular mass 73.2 kda). the amino acid sequence derived from cgh i showed high homology to a widely distributed family of beta-glucohydrolases (glycosyl hydrolases family 1). the recombinant cgh i protein produced in escherichia coli had cgh i activity. cgh i mrna was detected in leaves, flowers, stems and fruits of d. lanata.200011092916
cardenolide 16'-o-glucohydrolase from digitalis lanata. purification and characterization.a three-step chromatographic procedure was developed for purification of cardenolide 16'-o-glucohydrolase (cgh) from digitalis lanata ehrh. leaves, including phenyl-sepharose hydrophobic interaction chromatography followed by sp-sepharose cation exchange and q-sepharose anion-exchange chromatography. starting with acetone dry powder the purification resulted in an 760-fold enrichment of cgh. molecular weight, substrate specificity, ph optimum and temperature stability of cgh were determined. ant ...19989640670
cell type and spatial location dependence of cytoplasmic viscosity measured by time-resolved fluorescence microscopy.information on the cell type and spatial location dependence of cytoplasmic viscosity would be very useful in understanding some of the processes occurring in the cell. for this purpose, fluorescent dye kiton red (sulforhodamine b) was loaded into a variety of cells such as swiss 3t3 fibroblasts, human mononuclear cells, sarcoma-180 tumor cells, chinese hamster ovary cells, plant cells from digitalis lanata, stamen hair cells of tradescantia, and guard mother cells of allium cepa. space-resolved ...19979143317
[biological and chemical evaluations of extracts of digitalis lanata e. and digitalis purpurea l]. 19675613031
antiherpes activity of glucoevatromonoside, a cardenolide isolated from a brazilian cultivar of digitalis lanata.cardiac glycosides, known ligands of the sodium pump, are widely used in the treatment of heart failure, such as digoxin and digitoxin. besides this important activity, other biological activities, such as the antiviral activity, have been described for this group. hsv are responsible for many infections of oral, ocular and genital regions. treatment with nucleoside analogs such as acyclovir is effective in most cases; however drug-resistance may arise due to prolonged treatment mainly in immuno ...201121763352
biotransformation of extracted digitoxin from digitalis lanata by streptomyces.the biotransformation of digitoxin and some of its derivatives extracted from digitalis lanata by streptomyces isolated species was investigated. cultures of a streptomyces strain designated eusa2003b, isolated from an egyptian soil sample, efficiently induced selective 12beta-hydroxylation of the steroid aglycone of digitoxin (dt) and its alpha-acetyl and beta-methyl derivatives. the transformation reaction was performed within a 5-day fermentation process, products were isolated and their agly ...201121699088
[steroid saponins with more than one sugar chain. vi. lanatigoside and lanagitoside, two bisdesmosidic 22-hydroxyfurostanol glycosides from the leaves of digitalis lanata ehrh]. 20165077133
[mosaic virus on digitalis lanata ehrh. and its effect on the content of the active agent]. 196514309862
selective cytotoxicity evaluation in anticancer drug screening of fractionated plant extracts.chosen to reflect biodiversity in a phylogenetic sense, 100 fractionated plant extracts were screened in vitro for cytotoxicity following extraction and fractionation (polypeptide isolation). of these 100 extracts, 30 were selected and then characterized preliminarily for antitumor potency and mode of action by testing them on two cell lines and primary cultures of human tumor cells. on the basis of cytotoxicity potency, 10 of the extracts were further characterized for anticancer activity in 10 ...200212230887
optimization and validation of a high-performance liquid chromatography method for the analysis of cardiac glycosides in digitalis lanata.in this study, a simple and reliable hplc method for the qualitative and quantitative analysis of cardiac glycosides in digitalis lanata ehrh. raw material was developed and applied to healthy and phytoplasma-infected plants. the target analytes cover a broad range of secondary metabolites, including primary, secondary and tertiary glycosides and the corresponding aglycones. the sample preparation was carried out by sonication of the plant material with 70% (v/v) aqueous methanol at room tempera ...200919268961
stress-induced expression of cyclophilins in proembryonic masses of digitalis lanata does not protect against freezing/thawing stress.using proembryonic masses (pems) of digitalis lanata erh., it was demonstrated that cold, hormonal or osmotic stress, which increased freezing tolerance during cryopreservation, induced an increasing level of two peptidyl-prolyl-cis/transisomerases (ppiases). the difference in pi (9.2 +/- 0.2 and 9.5 +/- 0.2, +/- sd; n = 3) allowed the separation of the two enzymes by free-flow isoelectrophoresis. both were inhibited by cyclosporin a and thus belong to the cyclophilin family of ppiases. the enzy ...199910420652
quantitative determination of cardiac glycosides in digitalis lanata leaves by reversed-phase thin-layer chromatography.an analytical method for the determination of cardiac glycosides in digitalis lanata leaves by reversed-phase thin-layer chromatography (rp-tlc) was developed. the procedure consisted of extraction of dry leaf powder with 50% methanol and clean-up by sep-pak cartridges prior to rp-tlc analysis. rp-tlc was performed on an octadecylsilyl bonded silica gel plate, using a developing solvent of acetonitrile-methanol-0.5 m nacl (1:1:1) for primary glycosides and acetonitrile-methanol-0.5 m nacl (12:7: ...19968916557
extraction and quantitation of digoxin and acetyldigoxin from the digitalis lanata leaf via near-supercritical methanol-modified carbon dioxide.an extraction process is reported that employs a near-supercritical mixture of co2 and meoh to extract the cardiac glycoside, digoxin, from the digitalis lanata leaf. the method development of the sample preparation procedure is presented in detail, and reasons for trends that occur in the natural products extraction are given.19968759168
drugs for atrial fibrillation. digoxin comes from digitalis lanata. 19968611904
cardiac glycosides in partly submerged shoots of digitalis lanata.shoot cultures were established from axillary buds (11 strains) or seeds (1 strain) of individual digitalis lanata ehrh. plants and propagated partially submerged in liquid medium. five of these shoot culture strains were characterized with regard to their growth and cardenolide content. the cultures were observed for more than one year and found to be relatively stable with regard to their growth and cardenolide spectrum and yield. the strains examined differed in terms of their total cardenoli ...19938302954
biotransformation of 21-o-acetyl-deoxycorticosterone by cell suspension cultures of digitalis lanata (strain w.1.4).cell cultures of digitalis species are known to accept exogenous substrates for biotransformation reactions. we here report the biotransformation of 21-o-acetyl-deoxycorticosterone (1) by cell suspension cultures of digitalis lanata strain w.1.4. nine derivatives of 1 were obtained and their chemical structures determined by spectroscopic methods. 2β-hydroxylation and c-21-glucosylation of the steroidal nucleus were described for the first time in suspension-cultured plant cells. steroid 5α- and ...201222917633
expression of 3beta-hsd and p5betar, genes respectively coding for delta5-3beta-hydroxysteroid dehydrogenase and progesterone 5beta-reductase, in leaves and cell cultures of digitalis lanata ehrh.plants of the genus digitalis produce 5 beta-cardenolides that are used in the therapy of cardiac insufficiency in humans. 3 beta-hydroxysteroid dehydrogenase (3 beta-hsd) and progesterone 5 beta-reductase (p5 betar) are both supposed to be important enzymes in the biosynthesis of these natural products. activity and gene expression were demonstrated for both enzymes in cardenolide-accumulating leaves of digitalis lanata but also in cardenolide-free permanent cell suspension cultures initiated f ...201020514608
separation of cardiac glycosides by micellar electrokinetic capillary electrophoresis.the separation of mixtures of primary and secondary cardiac glycosides by micellar electrokinetic capillary electrophoresis modified by cyclodextrins, urea and sodium cholate proved to be suitable for the determination of these hydrophobic compounds. it was possible to distinguish the two anomeric cardenolides glucodigifucoside and glucodigiglucomethyloside with all three buffer systems. electropherograms of crude plant cell extracts from digitalis lanata were obtained with this method.19938486748
high-performance liquid chromatographic determination of cardenolides in digitalis leaves after solid-phase extraction.for hplc analysis of cardenolide glycosides of digitalis lanata the separation of the main compounds from other constituents is useful. an improved method for doing this, based on solid-phase extraction on a c18 modified poly(styrene-divinylbenzene) polymer, is described. the presented assay permits quantitative estimation of more than 50 cardenolides in about 2 mg of dried leaf powder of digitalis lanata with high speed and accuracy.19938445005
delta 5-3beta-hydroxysteroid dehydrogenase (3 beta hsd) from digitalis lanata. heterologous expression and characterisation of the recombinant enzyme.during the biosynthesis of cardiac glycosides, delta (5)-3beta-hydroxysteroid dehydrogenase (3 beta hsd, ec 1.1.1.51) converts pregnenolone (5-pregnen-3beta-ol-20-one) to isoprogesterone (5-pregnene-3,20-dione). a 3 beta hsd gene was isolated from leaves of digitalis lanata. it consisted of 870 nucleotides containing a 90 nucleotide long intron. a full-length cdna clone that encodes 3 beta hsd was isolated by rt-pcr from the same source. a sph i /kpn i 3 beta hsd cdna was cloned into the pqe30 v ...200717564944
effects of digitoxigenin, digoxigenin, and various cardiac glycosides on cardenolide accumulation in shoot cultures of digitalis lanata.various cardenolide genins and cardenolide glycosides were administered to light-grown and dark-grown digitalis lanata shoot cultures to investigate conversion reactions related to the formation and rearrangement of the sugar side chain of digitalis glycosides. digitoxigenin was converted to digitoxigen-3-one, 3-epidigitoxigenin, and digoxigenin. in addition, various cardiac glycosides were formed, including mono-glycosides with glucose, glucomethylose, fucose, and digitalose, as well as the cor ...199817253315
effects of various pregnanes and two 23-nor-5-cholenic acids on cardenolide accumulation in cell and organ cultures of digitalis lanata.5-pregnen-3beta-ol-20-one (pregnenolone), 4-pregnene-3,20-dione (progesterone), 5-pregnene-3beta,21-diol-20-one (21-hydroxypregnenolone), 4-pregnen-21 -ol-3,20-dione (cortexone), 5beta-pregnane-3,20-dione, 5alpha-pregnane-3,20-dione, 5beta-pregnan-3alpha-ol-20-one, 5beta-pregnan-3beta-ol-20-one, 5beta-pregnane-3beta,14beta, 21-triol-20-one 3-acetate, 23-nor-5-cholenic acid-3beta,20xi-diol, and 23-nor-3,20(22) e-choladienic acid-3beta-ol were administered to photomixotrophic shoot cultures of dig ...199717252366
crystallization and preliminary crystallographic analysis of selenomethionine-labelled progesterone 5beta-reductase from digitalis lanata ehrh.progesterone 5beta-reductase (5beta-por) catalyzes the reduction of progesterone to 5beta-pregnane-3,20-dione and is the first stereospecific enzyme in the putative biosynthetic pathway of digitalis cardenolides. selenomethionine-derivatized 5beta-por from d. lanata was successfully overproduced and crystallized. the crystals belong to space group p4(3)2(1)2, with unit-cell parameters a = 71.73, c = 186.64 a. a mad data set collected at 2.7 a resolution allowed the identification of six out of e ...200616511297
use of commercial plant species in a hydroponic system to treat domestic wastewaters.the objectives in this work were to investigate a conceptual layout for an inexpensive and simple system that would treat primary municipal wastewater to discharge standards. a commercial hydroponic system was adapted for this study and the wastewater was used to irrigate wooly digitalis (digitalis lanata ehrh.) and foxglove (digitalis purpurea l.). these plants are medicinal and produce cardenolide compounds. influent and effluent samples were collected once a month for six months and analyzed ...201315074822
[preparation of biologically transformed raw material from woolly foxglove digitalis lanata ehrh and isolation of digoxin from it].a biotechnological approach is proposed for conservation of a terraneous part of woolly foxglove under anaerobic conditions with a subsequent air-sun drying of the biologically transformed raw material. during the conservation primary foxglove glycosides completely convert to secondary ones which do not transform further. a simple method is described for preparation from the transformed raw material of an enriched glycoside fraction with the yield of 3.6% and for isolation from this fraction of ...201314593875
[solubility and dissolution rate of digoxin from digitalis lanata drug extracts] .the influence on solubility and dissolution rate was investigated for digoxin as a model drug with a very low solubility in water. the investigations were carried out with different fractions of extracts from leaves of digitalis lanata. these fractions differ in the composition of concomitant compounds. the solubility of digoxin from the extract fractions is increased up to 42 times, with considerable differences between the fractions. the solubility depends on the weight of the extract fraction ...200011189870
haploid plants regenerated from androgenic cell cultures of digitalis lanata.androgenic callus was obtained from cold treated anthers and pollen of digitalis lanata. the callus was mixoploid and contained haploid, diploid and tetraploid cells as shown by impulse cytophotometry. haploid cell lines were selected by colony cloning. they were unstable and selection had to be repeated every 1-2 months. mixoploid shoot cultures were derived from embryogenic haploid cell lines via somatic embryos. haploid shoots were selected by explanting shoot tips. the shoots showed wide var ...200010821049
structure-specificity relationship of cardiac glycosides as a substrate for glucohydrolase ii.cardenolide glucohydrolase ii (cgh ii) is a cardenolide-specific glucohydrolase obtained from digitalis lanata leaves. we investigated the structure-specificity relationship of several cardenolide disaccharides as a substrate for cgh ii. conformation analysis of the substrates was performed using molecular mechanics calculations. the sugar chain conformation of two inert glycosides was significantly different from that of the other glycosides. the other two glycosides, which were weak substrates ...200010726855
purification of δ(5)-3-ketosteroid isomerase from digitalis lanata.the isomerization of 5-pregnene-3,20-dione into 4-pregnene-3,20-dione was investigated to shed further light on cardenolide biosynthesis and to characterize the enzymes involved in cardenolide formation. it was shown that the δ(5)-3-ketosteroid isomerase of digitalis lanata, which catalyzes this isomerization, is an individual enzyme and not, as previously thought, associated with δ(5)-3β-hydroxysteroid dehydrogenase. the enzyme was purified by fractionated ammonium sulfate precipitation, hydrop ...201525468533
prospecting endophytic fungal assemblage of digitalis lanata ehrh. (foxglove) as a novel source of digoxin: a cardiac glycoside.endophytes, the chemical synthesizers inside plants, are the microorganisms having mutualistic relationship with the host plant. they can be used by plants for defense in addition to the production of a wide variety of beneficial bioactive secondary metabolites. there are reports that microbial endophytes mimic the bioactive compounds as produced by the plant itself thus making them a promising source of novel compounds. during the present study, endophytes were isolated from the symptomless lea ...201328324591
[effect of some biotic elicitors on flavonoids production in digitalis lanata cell cultures].a method to improve the secondary metabolites production in plant cell cultures is the use of elicitors (biotic or abiotic). in this paper we investigated the effect of two biotic elicitors, botrytis and sclerotinia, on the flavonoids production, an important category of secondary metabolites, in digitalis lanata suspension cultures.201323077964
lanatoside c, a cardiac glycoside, acts through protein kinase cδ to cause apoptosis of human hepatocellular carcinoma cells.recent studies have revealed that cardiac glycosides, such as digitalis and digoxin, have anticancer activity and may serve as lead compounds for the development of cancer treatments. the poor prognosis of hepatocellular carcinoma (hcc) patients reflects the development of resistance to current chemotherapeutic agents, highlighting the need for discovering new small-molecule therapeutics. here, we found that lanatoside c, an anti-arrhythmic agent extracted from digitalis lanata, inhibited the gr ...201728387249
cultivation and breeding of digitalis lanata in the netherlands.after the second world war marshall aid funds were used to establish a cooperative organisation for growing, drying, and selling digitalis lanata (and other medicinal, aromatic, and culinary herbs) in the netherlands. the crop is sown in mid april and the fully mechanised harvest of the leaves takes place from september to late november. the leaves are dried for 10-12 hours at 50 degrees c maximum. the aim of breeding trials is to improve leaf production, erect leaf attitude, resistance to septo ...19854041298
structural studies on the biosides of digitalis lanata: bisdigitoxosides of digitoxigenin, gitoxigenin and digoxigenin.the crystal structures and conformations of bisdigitoxosides of digitoxigenin (i), gitoxigenin (ii) and digoxigenin (iii and iv) have been determined using single-crystal x-ray crystallographic techniques. crystals of (i), (ii) and (iv) were grown from ethyl acetate solutions of the glycosides while (iii) was grown from a solution of the digitoxoside in ethanol. as in other cardiac glycosides the ring junctions a-b and c-d are cis. the d ring in these structures shows different conformations whi ...19892619961
studies on growth and cardenolide production of digitalis lanata tissue cultures.digitalis lanata cell cultures grown as small undifferentiated aggregates in suspension culture can be redifferentiated into green embryos that produce cardenolides. the possibility of using a statistical (box-wilson) experimental design to study the effects of four different variables on growth, differentiation, and cardenolide production of d. lanata tissue cultures are investigated. the results of the analyses were processed by linear regression analysis. mathematical models explaining the ef ...198918587951
purification and characterization of lanatoside 15'-o-acetylesterase from digitalis lanata ehrh.lanatoside 15'-o-acetylesterase (lae) from in-vitro-cultivated cells of digitalis lanata ehrh. was isolated and partially sequenced. the enzyme was extracted with citrate buffer from acetone dry powder. it was purified in a two-step chromatographical procedure including phenyl sepharose hydrophobic interaction chromatography followed by cm sepharose cation-exchange chromatography to more than 330 mumol.s-1.(g protein)-1. sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page) of the ...19989530881
extracellular polysaccharides produced by suspension-cultured cells from digitalis lanata.extracellular polysaccharides (ecp) were isolated in yields of up to 4 mg/ml from the culture media of suspension-cultured cells from digitalis lanata ehrh. ecp content was increasing continuously over the first ten days of cultivation and then stayed constant until day 20. ecp were fractionated by ion-exchange chromatography into two neutral and one acidic fractions. further fractionation was achieved by gel-permeation chromatography (gpc). one neutral fraction was separated into two distinct f ...19979342950
21'-di-dehydro-deacetyllanatoside c, a biotransformation product of deacetyllanatoside c from senescent shoot cultures of digitalis lanata.feeding deacetyllanatoside c to senescent shoot cultures of digitalis lanata resulted in the formation of a new product, which was isolated by semi-preparative hplc. the molecular structure was elucidated by means of hplc-mass spectrometry and nmr as 21'-di-dehydro-deacetyllanatoside c.19979055448
isolation of cardenolides from a brazilian cultivar of digitalis lanata by rotation locular counter-current chromatography.cardenolides from a brazilian cultivar of digitalis lanata were isolated by rotation locular countercurrent chromatography (rlcc), employing dichloromethane-methanol-water (5:6:4, v/v) as the solvent system. highly pure lanatoside c was obtained from the digitalis lanata hydromethanolic extract, pre-purified either by silica gel or reversed-phase chromatography.19969008858
[determination of cardenolides in digitalis lanata by a solid-phase immunoenzyme method].a competitive solid-phase eia technique using high-affinity digoxin-specific monoclonal antibodies has been developed to determine cardenolides in digitalis lanata. the test-system can detect digoxin and closely related compounds in concentration of 0.1 nmol/ml. the content of cardenolides, tested by elisa, correlated well with the total content of digoxin, deslanoside c and lanatoside c as analysed by hplc. the test-system is useful in estimating the productivity of large series and samples of ...19938274170
3 alpha-hydroxysteroid-5 beta-oxidoreductase in tissue cultures of digitalis lanata.putative intermediates of cardenolide biosynthesis, namely progesterone, pregnenolone, 5 beta-pregnane-3,20-dione or 5 beta-pregnan-3 beta-ol-20-one, were administered to light- or dark-grown shoot cultures of digitalis lanata. the unsaturated compounds were reduced to their respective 5 alpha-pregnanes, 5 beta-pregnane-3,20-dione was reduced to 5 beta-pregnan-3 alpha-ol-20-one and 5 beta-pregnan-3 beta-ol-20-one was isomerized to the respective 3 alpha-pregnane. suspension cultures of digitalis ...20138251035
12 beta-hydroxylation of digitoxin by suspension-cultured digitalis lanata cells: production of digoxin in 20-litre and 300-litre air-lift bioreactors.a biotransformation process for the production of digoxin was developed using digitalis lanata cell suspension cultures. digitoxin was used as the substrate for biotransformation. digoxin production was carried out in a variety of vessels, including 1-l exsiccators, 20-l glass reactors and a 300-l air-lift bioreactor. a culture volume of 200 l was established after 28 d and the cells were then cultured semi-continuously in a 300-l bioreactor employing the draw-fill cultivation method. maximal di ...19921369154
cardenolide and neutral lipid biosynthesis from malonate in digitalis lanata.digitalis lanata plants were grown on water culture in a controlled environment and in the young, growing leaves free sterols (0.335 micromol/g fw), triacylglycerols (0.97 micromol/g fw) and cardenolides (1.82 micromol/g fw) were the major apolar and polar lipids. the cardenolide-containing fraction from these tissues was separated into 26 cardenolides by hplc. the 5 major components (accounting for 60 % of the occurring glycosides) lanatosides a and c, acetyldigoxin, acetyldigitoxin, and glucoe ...199217226464
major cardenolide glycosides in embryogenic suspension cultures of digitalis lanata. 19873628562
udp-glucose:digitoxin 16'-o-glucosyltransferase from suspension-cultured digitalis lanata cells.suspension cultures from several cell lines of digitalis lanata, as well as cultures from 6 other plant species were checked for their ability to form purpurea-glycoside a from digitoxin. an in-vitro assay for the udp-glucose:digitoxin 16'-o-glucosyltransferase (dgt, ec 2.4.1.-) has been established based on an hplc method. the enzyme is located in the soluble fraction. its ph optimum is at 7.4. no enzyme activity was found in either purified vacuole preparations or lysed vacuoles. ascorbate (10 ...198624248401
co-cultivation of digitalis purpurea and digitalis lanata cell cultures. 198617345441
long-term cultivation of digitalis lanata clones propagated in vitro: cardenolide content of the regenerated plants1.shoot cultures of digitalis lanata ehrh. (scrophulariaceae) were established by inoculating meristem explants from axillary buds in a halfstrength liquid murashige and skoog medium. after short- and long-term cultivation in vitro, the shoots were rooted on a solid medium, transferred into soil, and grown in the greenhouse. the cardenolide content of the regenerated plants was determined by hplc. the vegetatively propagated plants showed good homogeneity and are identical with the parent plant in ...198617345413
[cardenolide glycosides in digitalis crossbreeds. reciprocal crossbreeds of digitalis lanata with other digitalis species]. 19921642514
high-performance liquid chromatographic determination of alpha-acetyldigoxin in digitalis lanata leaves.an analytical method for the determination of alpha-acetyldigoxin in digitalis lanata leaves by hplc has been developed. the procedure consists of extraction of dry leaf powder with 50% methanol and cleanup by a sep-pak c18 cartridge prior to hplc analysis. the quantitation is carried out by the incorporation of beta-methyldigoxin as an internal standard. hplc is performed on an octylsilyl bonded silica column with acetonitrile/methanol/water (100/11/188, v/v). the effluent is monitored by uv ab ...19911776696
contamination of botanical dietary supplements by digitalis lanata. 19989738088
determination of cardenolides in hairy root cultures of digitalis lanata by enzyme-linked immunosorbent assay.a competitive elisa technique using digoxin-specific antibody has been developed to determine digoxin and its related compounds in hairy root cultures of digitalis lanata. the elisa could detect 0.2-2 nm digoxin and closely related cardenolides. hairy roots cultured in the dark accumulated very small amounts of cardenolides (0.02-0.07 micrograms/g dry wt), while the content of cardenolides in green hairy roots cultured in the light was increased maximally 600-fold (16.5 micrograms/g dry wt) comp ...20132089119
new cardiac glycosides from digitalis lanata.from the leaves of digitalis lanata ehrh. two new cardenolides have been found and structurally elucidated as digoxigenin-3-o-beta- d-digitoxosido-beta- d-digitoxosido-beta- d-2,6-dideoxy-glucoside and glucodigoxoside by tlc and spectroscopic methods.198417340309
new cardiac glucosides from digitalis lanata1.from the leaves of digitalis lanata ehrh. two as yet unknown cardenolides have been found. they were structurally elucidated as digoxigenin-3-o-beta- d-digitoxosido-beta- d-2,6-dideoxyglucoside and digoxigenin-3-o-beta- d-digitaloside by tlc and mainly spectroscopic methods.198417340285
stability of biotransformation capacity in digitalis lanata cell cultures after cryogenic storage.suspension cultures of digitalis lanata were stored at the temperature of liquid nitrogen. after thawing, viable cultures were recovered which showed a growth rate equal to that of untreated cells. the capacity of digitalis cells to transform β-methyldigitoxin to β-methyldigoxin remained unchanged after cryostorage. this was shown in 300-ml shake cultures as well as in 20-1 bioreactors. from these results it is clear that cryostorage is a suitable method for preserving cell lines with specific c ...198324258127
[new cardiac glycosides from digitalis lanata.].from the leaves of digitalis lanata e hrh. three as yet unknown cardenolides were isolated and structurally elucidated as digoxigenin-3-o-beta-d-digitoxosido-beta-d-digitoxosido-beta-d-glucomethylosid, digoxigenin-3-o-beta-d-digitoxosido-beta-d-glucomethylosid and diginatigenin-3-o-beta-d-digitalosid by tlc and mainly spectroscopic methods.198317405021
[peroxidase activity and peroxidase isoenzyme patterns of tissue cultures of digitalis lanata and digitalis purpurea.]. 198317404926
two-stage cultivation of digitalis lanata cells: semicontinuous production of deacetyllanatoside c in 20-litre airlift bioreactors.a two-stage cultivation method was employed to develop a semicontinuous biotransformation process for the production of deacetyllanatoside c, a cardenolide of the important digoxin series. digitoxin was used as the substrate for biotransformation. the process was optimized in 1-l shake flasks and then established on the 20-l scale using two airlift bioreactors, one for cell growth (working volume 12 litres) and another for deacetyllanatoside c production (working volume 18 litres). growth and pr ...19901366814
3 beta-hydroxysteroid oxidoreductase in suspension cultures of digitalis lanata ehrh.a 3 beta-hydroxysteroid oxidoreductase was isolated and characterized in the microsomes of digitalis lanata cell cultures. the enzyme catalyzes the conversion of 5 alpha-pregnane-3,20-dione to 5 alpha-pregnan-3 beta-ol-20-one and requires nad(p)h2. the enzyme was found to have a ph optimum of 8.0. the reaction had an optimum incubation temperature of 25 degrees c with linear reduction for the first 4 h, reaching maximum enzyme activity after 7 h. substrate kinetics for 5 alpha-pregnane-3,20-dion ...20092291772
separation and determination of lanatosides in digitalis lanata leaves by high-performance liquid chromatography. 19902380317
batch cultures of somatic embryos of digitalis lanata in gaslift fermenters. development and cardenolide accumulation.somatic embryos of digitalis lanata strain vii were successfully grown as batch cultures in gaslift fermenters. embryo development and formation of cardenolides in the embryos resembled those of cultures grown in shake flasks. both processes depended on the developmental stage of the embryos at the time point of inoculation, the homogeneity of the embryo structures, the density of the inoculum, the composition of the nutrient medium, the intensity of irradiation, and the composition of the gas m ...199017221393
light energy dissipation under water stress conditions: contribution of reassimilation and evidence for additional processes.using (14)co(2) gas exchange and metabolite analyses, stomatal as well as total internal co(2) uptake and evolution were estimated. pulse modulated fluorescence was measured during induction and steady state of photosynthesis. leaf water potential of digitalis lanata ehrh. plants decreased to -2.5 megapascals after withholding irrigation. by osmotic adjustment, leaves remained turgid and fully exposed to irradiance even at severe water stress. due to the stress-induced reduction of stomatal cond ...199016667370
δ5-3β-hydroxysteroid dehydrogenase/δ5-δ4-ketosteroid isomerase (3β-hsd), a possible enzyme of cardiac glycoside biosynthesis, in cell cultures and plants of digitalis lanata ehrh.the in vitro transformation of pregnenolone into progesterone in digitalis lanata tissues was shown to be catalyzed by a 3β-hydroxysteroid dehydrogenase/ketosteroid isomerase (3β-hsd). product formation was monitored by hplc. the enzyme could be partially characterized and 3β-hsd activities were measured in various digitalis lanata tissues and in cell cultures of other plant species. since no correlation was observed between biosynthetic competence of the tissue and 3β-hsd activity, it was concl ...199024232686
delta(5)-3beta-hydroxysteroid dehydrogenase from digitalis lanata ehrh. - a multifunctional enzyme in steroid metabolism?delta(5)-3beta-etaydroxysteroid dehydrogenase (delta(5)-3beta-hsd; ec 1.1.1.145), an enzyme converting pregn-5-ene-3beta-ol-20-one (pregnenolone) to pregn-5-ene-3,20-dione (isoprogesterone), was isolated from the soluble fraction of suspension-cultured cells of digitalis lanata l. strain viii. starting with acetone dry powder the enzyme was purified in three steps using column chromatography on fractogel-tsk deae, hydroxyapatite and sephacryl g-200. fractions with highest delta(5)-3beta-hsd acti ...199910550629
[cardiac glycosides: from ancient history through withering's foxglove to endogeneous cardiac glycosides].for centuries, drugs that increase the power of contraction of the failing heart have been used for the treatment of congestive heart failure (dropsy). the cardiac effect is due to the content of cardiac glycosides. squill or sea onion, urginea (scilla) maritima, a seashore plant, was known by the ancient romans and syrians and possibly also by the ancient egyptians. squills were used erratically, but some prescriptions indicate that they may have been used for the treatment of oedematous states ...200415685783
characterization and localization of progesterone 5 alpha-reductase from cell cultures of foxglove (digitalis lanata ehrh).progesterone 5 alpha-reductase, which catalyses the reduction of progesterone to 5 alpha-pregnane-3,20-dione, was isolated and characterized from cell cultures of digitalis lanata (foxglove). optimum enzyme activity was observed at ph 7.0, and the enzyme had an apparent km value of 30 microm for its substrate progesterone. the enzyme needs nadph as reductant, which could not be replaced by nadh. for nadph, the apparent km value is 130 microm. the optimum temperature was 40 degrees c; at temperat ...19902106876
formation of digitalis lanata clone lines by shoot tip culture.propagation of digitalis lanata by shoot tip culture made possible (a) the rapid multiplication of elite plants with the formation of plant clones and (b) the long-term cultivation of these plants which under normal growth conditions would die at the end of the second vegetation period. optimum conditions were established for the regeneration of shoots from shoot tips, for daughter shoot formation and rooting as well as for the adaptation of the regenerated plants to the open ground. gene banks ...199017221370
semicontinuous cultivation of digitalis lanata cells: production of beta-methyldigoxin in a 300-l airlift bioreactor.selected digitalis lanata cell lines cultivated in 1-l shake flasks or 20-l airlift bioreactors converted beta-methyldigitoxin into beta-methyldigoxin with almost no side reactions. this biotransformation process was optimized with regard to substrate supply and culture medium composition, and was then scaled up to a volume of 210 l using a 300-l airlift bioreactor. a semicontinuous process was developed in which 513.3 g beta-methyldigoxin were produced after 89 days of cultivation.198918588131
the efficiency of water use in water stressed plants is increased due to aba induced stomatal closure.gas exchange and abscisic acid content of digitalis lanata ehrh. have been examined at different levels of plant water stress. net photosynthesis, transpiration and conductance of attached leaves declined rapidly at first, then more slowly following the withholding of irrigation. the intercellular partial pressure of co2 decreased slightly. the concentration of 2-cis(s)aba increased about eight-fold in the leaves of non-irrigated plants as compared with well-watered controls. a close linear corr ...198824425243
growth and digoxin content in digitalis lanata in controlled conditions and natural environment. 19883174863
reconstitution of cytochrome p-450-dependent digitoxin 12 beta-hydroxylase from cell cultures of foxglove (digitalis lanata ehrh.).cytochrome p-450-dependent digitoxin 12 beta-hydroxylase from cell cultures of foxglove (digitalis lanata) was solubilized from microsomal membranes with chaps (3-[(3-cholamidopropyl)dimethylammonio]propane-1-sulphonic acid). cytochrome p-450 was separated from nadph: cytochrome c (p-450) reductase by ion-exchange chromatography on deae-sephacel. nadph:cytochrome c (p-450) reductase was further purified by affinity chromatography on 2',5'-adp-sepharose 4b. this procedure resulted in a 248-fold p ...19883137929
rapid clonal multiplication of digitalis lanata in tissue culture.shoot cultures of digitalis lanata have been established by inoculating the shoot tip of seedlings germinated in aseptic culture, or of field-grown plants, onto linsmaier and skoog's rm medium supplemented with 1 mg 6-benzylaminopurine and 0.1 mg indolacetic acid 1(-1). on this medium formation of up to 30 new axillary shoots could be induced. shoots could be grown into functional plants after root induction on a medium containing reduced amounts (one-fifth of normal) of nitrogen and indolebutyr ...198124258753
12beta-hydroxylation of digitoxin by suspension-cultured digitalis lanata cells. production of deacetyllanatoside c using a two-stage culture method1.suspension-cultured digitalis lanata cells, known to form beta-methyldigoxin from beta-methyldigitoxin without any by-products, were not able to 12beta-hydroxylate digitoxin efficiently when cultivated in the cell culture medium devised by murashige and skoog. most of the substrate added was merely glucosylated at its 16'-o-position leading to purpureaglycoside a as the main biotransformation product after 9 days of incubation. an 8% glucose solution (ph 5.5) turned out to be a suitable producti ...198817265224
fluorescence quenching and gas exchange in a water stressed c(3) plant, digitalis lanata.a leaf cuvette has been adapted for use with a pulse-modulation fluorometer and an open gas exchange system. leaf water potential (psi) was decreased by withholding watering from digitalis lanata ehrh. plants. at different stages of water deficiency the photochemical (q(q)) and nonphotochemical (q(e)) fluorescence quenching was determined during the transition between darkness and light-induced steady state photosynthesis of the attached leaves. in addition, the steady state co(2) and h(2)o gas ...198816665875
cryopreservation of digitalis lanata shoot tips.a method for the preservation in liquid nitrogen of shoot tips (meristems) of d. lanata is described. it includes the following steps: (a) hardening of shoots by cultivation at 4 degrees c for 8 weeks, (b) treatment of the explanted shoot tips with cryoprotectors, e.g., 2 mol dmso l (-1) for 2 h, (c) either ultrarapid cooling (ca. 4000 k min (-1)) of the shoot tips by submerging in liquid nitrogen or slow cooling (ca. 0.5 k min (-1)) of the shoot tips to -40 degrees c using a suitable freezer, ( ...198717269043
immobilization of digitoxin 12β-hydroxylase, a cytochrome p-450-dependent enzyme from cell cultures of digitalis lanata ehrh.attempts were made to immobilize digitoxin 12β-hydroxylase, a membrane-bound, cytochrome p-450-dependent monooxygenase from cell cultures of digitalis lanata. the optimum procedure was the entrapment of microsomes in 2% alginate by crosslinking the polysaccharide chains with cacl2. after the immobilization of the enzyme about 70% of its activity was retained. the kinetic data such as the ph optimum and the optimum substrate concentrations were identical for the immobilized enzyme and freely susp ...198724248652
highly efficient 12beta-hydroxylation of digitoxin in digitalis lanata cell suspensions using a two-staged culture method. 198617345373
isolation and quantitative determination of some cardioactive glycosides from digitalis lanata by high-performance liquid chromatography.a rapid extraction method followed by high-performance liquid chromatographic assay was developed for the quantitative determination of the cardioactive glycosides of digitalis lanata. the leaf samples were extracted with water or aqueous alcohols. the simple extraction method gives a better yield than the methods described previously. lanatoside c and its metabolites have been separated on a reversed-phase column with various mixtures of acetonitrile, methanol, and water as mobile phases for is ...19863740407
[cultivation and improvement of digitalis lanata in the netherlands]. 19853990875
new cardiac glycosides from digitalis lanata.from the leaves of digitalis lanata ehrh. two new cardenolides have been found and structurally elucidated as digoxigenin-3-o-beta- d-digitoxosido-beta- d-digitoxosido-beta- d-xyloside bei tlc and spectroscopic methods.198417340310
clinical utility of plasma digoxin measurements.the results of plasma digoxin concentration (pdc) measurements in 260 patients treated with digitalis lanata preparations (digoxin, lanatoside c, betamethyl-digoxin) allowed the following observations: 1. owing to an important overlap between "toxic" and "nontoxic" pdc values, the method has a limited value in the diagnosis of digitalis toxicity. however, in patients with toxicity associated with pdc under 2 ng/ml, it allows identification digitalis sensitivity. 2. a dosage selection based on pd ...20132749155
biotransformation of secondary glycosides in digitalis lanata leaf discs. 198117402071
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