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At least 37 records · Page 2Linked to original sources
PubMed2000

Tropane alkaloid production by hairy roots of Atropa belladonna obtained after transformation with Agrobacterium rhizogenes 15834 and Agrobacterium tumefaciens containing rol A, B, C genes only.

Atropa belladonna leaf disks were infected by a wild strain Agrobacterium rhizogenes 15834 harboring the Ri-TL-DNA and by a disarmed Agrobacterium tumefaciens strain harboring a construction with only rol ABC and npt II genes. Thirteen root lines were established and examined for their growth rate and alkaloid productivity to evaluate the possible role of rol genes in morphological differentiation and in tropane alkaloid formation. A great diversity has been observed in the growth rate of these 13 root lines. The root biomass increased up to 75 times. The total alkaloid contents were similar in the root lines obtained by infection with A. rhizogenes 15834 and A. tumefaciens rol ABC. The last ones accumulated between 4 (1.1 mg g(-1) DW) and 27 (8 mg g(-1) DW) times more alkaloids than the intact roots (0.3 mg g(-1) DW). This work has shown that the rol ABC genes were sufficient to increase tropane alkaloid production in A. belladonna hairy root cultures.

Agrobacterium tumefaciens↗
PubMed2003

Alkaloids in plants and root cultures of Atropa belladonna overexpressing putrescine N-methyltransferase.

Putrescine N-methyltransferase (PMT) is the first alkaloid-specific enzyme for nicotine and tropane alkaloid formation. The pmt gene from Nicotiana tabacum was fused to the CaMV 35S promoter and integrated into the Atropa belladonna genome. Transgenic plants and derived root cultures were analysed for gene expression and for levels of alkaloids and their precursors. Scopolamine, hyoscyamine, tropine, pseudotropine, tropinone, and calystegines were found unaltered or somewhat decreased in pmt-overexpressing lines compared to controls. When root cultures were treated with 5% sucrose, calystegine levels were elevated in control roots, but were not affected in pmt-overexpressing roots. 1 microM auxin reduced calystegine levels in control roots, while in pmt-overexpressing roots all alkaloids remained unaltered. Expression level of pmt alone is apparently not limiting for tropane alkaloid formation in A. belladonna.

Atropa belladonna↗
PubMed2004

Comparison of the inhibitory efficacy of four belladonna drugs on gastrointestinal movement and cognitive function in food-deprived mice.

Belladonna drugs, scopolamine (Sco), atropine (Atr), anisodine (AT(3)), and anisodamine (Ani), frequently used for gastrointestinal motility disorders often produce adverse effects on the central nervous system. In the present work, these drugs (0.05, 0.5, 5, and 50 micromol kg(-1), i.p.) were evaluated for their potential to inhibit gastrointestinal motility and cognition in mice. Results showed that the maximum inhibitory rates of Sco, Atr, AT(3), and Ani on gastric emptying and small intestinal movement were 29.78, 40.69, 12.30, and 17.99% and 51.98, 58.46, 46.51, and 46.22%, respectively. The affinities of Sco, Atr, AT(3), and Ani for muscarinic receptors in the whole mice were 1.62, 1.48, 2.28, and 1.11 micromol kg(-1) for the stomach or 0.30, 1.12, 0.59, and 1.14 micromol kg(-1) for the small intestine, respectively. The minimal effective doses for impairing avoidance-response learning were 5 micromol kg(-1) for Sco, Atr, or AT(3) and 50 micromol kg(-1) for Ani. The initial doses for insulting the avoidance-response memory or open-field memory were 0.5, 5, 5, and 50 micromol kg(-1) or 5, 5, 5, and >50 micromol kg(-1) for Sco, Atr, AT(3), and Ani, respectively. We conclude that the relative susceptibility of the mouse's tissue or function capacities towards the inhibitory effects of belladonna drugs is small intestine > stomach > avoidance-response memory > avoidance-response learning > open-field memory.

Animals↗
PubMedDate not supplied

Characteristics of Atropa belladonna hairy roots cryopreserved by vitrification method.

Atropa belladonna hairy roots (clone M8) were successfully cryopreserved by using the vitrification method. A. belladonna hairy root tips were precultured on a half strength of Murashige and Skoog (MS) solid medium with 0.1 mg per L 2,4-D or without phytohormone for 1 day, and then dehydrated with PVS2 solution for 15 minutes prior to immersion into liquid nitrogen for 1 day, 1 week, 1 month and 3 months. Hairy root tips kept in liquid nitrogen were rapidly thawed at 36 degree C in a water bath. The root tips were recultured on half strength MS medium. The hairy root tips, precultured with 2,4-D before cryopreservation, showed a higher survival rate than those precultured without phytohormone. The hairy root tips, precultured with 2,4-D, showed an average survival rate of 83 percent. There was no significant difference in the viability of the hairy roots cryopreserved for different periods. The regrowth of cryopreserved hairy roots was similar to that of untreated hairy roots and tropane alkaloid productivity became stable after 4th subculture. PCR analysis of hairy roots demonstrated the conservation of the T-DNA in cryopreserved hairy roots. These results indicate that cryopreservation by vitrification method is useful to preserve A.belladonna hairy root clone M8.

Atropa belladonna↗
PubMed1982

Belladonna alkaloids and phenobarbital combination pharmaceuticals analysis II: High-performance liquid chromatographic determination of phenobarbital.

A high-performance liquid chromatographic separation is described for the analysis of phenobarbital in combination pharmaceutical dosage forms containing belladonna alkaloids. A mobile phase of 0.003 M tetramethylammonium chloride in water-methanol (3:2, pH 7.4) was used to separate phenobarbital from belladonna alkaloids on an octadecylsilane column in less than 7 min. The column effluent was monitored at 240 nm, which resulted in a detection limit of 6 ng of phenobarbital. The method is applicable to elixirs, tablets, and capsules with no preliminary extraction procedure. Data from the application of the method to commercial products is also presented.

Belladonna Alkaloids↗
PubMed1993

Does a highly diluted homoeopathic drug act as a placebo in healthy volunteers? Experimental study of Belladonna 30C in double-blind crossover design--a pilot study.

A basic tenet of homoeopathy is that remedies which do not contain active molecules can have effects on the healthy human organism, by virtue of the specific preparation process of stepwise dilution and succussion, called potentization. The claim that a so called high 'potency' of a homoepathic remedy, Belladonna C30, could produce effects different from placebo, was investigated in a pilot study. In a double-blind crossover trial, 4 weeks of Belladonna C30 were compared to 4 weeks of placebo in 47 healthy volunteers. Data were collected daily. The number and types of changes were recorded into a predefined category system. Single-case evaluation showed differences between the two experimental phases for 21 subjects. Group evaluation showed no clearcut differences. The claim that homoeopathic potencies can produce symptoms other than placebo in healthy subjects should be put to further scrutiny.

Adult↗
PubMed2001

Neurotropic, immunological and gastric effects of low doses of Atropa belladonna L., Gelsemium sempervirens L. and Poumon histamine in stressed mice.

Previous studies realized in the laboratory have indicated that application of experimental stress (such as unavoidable footshock) induced significant behavioral, gastric and immunological alterations in mice. The aim of this study was to evaluate effects of low doses of Atropa belladonna L., Gelsemium sempervirens L. and Poumon histamine on stress-induced behavioral, immunological and gastric alterations. Locomotor, postural and exploratory activities have been evaluated by two behavioral tests: light/dark box and staircase tests. Immunological studies were investigated to count white blood cells subpopulations (lymphocytes, neutrophils, monocytes and basophils) by coulter counter. The severity of gastric erosions was evaluated by microscopic technique in mice after experimental stress. The results have demonstrated that low doses of G. sempervirens L. and A. belladonna L. had a significant neurotropic and protective effects on behavioral and gastric alterations induced by experimental stress. The immunological protective effects observed were probably induced via their neurotropic effects. The P. histamine showed a significant immunoprotective and gastroprotective effect in mice exposed to experimental stress.

Alkaloids↗
PubMed2000

Effects of the rol C gene on hairy root: induction development and tropane alkaloid production by Atropa belladonna.

Two series of Atropa belladonna hairy root lines were obtained, the first one transformed via Agrobacterium tumefaciens harboring rol C and npt II genes, and the other transformed with rol ABC and npt II genes. Thirteen hairy root lines were obtained and selected on hormone-free medium. The transformation was confirmed by PCR analysis, and these root lines were first examinated for their growth rate. Then hyoscyamine and scopolamine production was measured after 3 and 4 weeks of culture to evaluate the possible role of rol C gene in tropane alkaloid formation. The rol C gene alone played a significant role in the hairy root growth rate (17-fold increase). However this effect was much lower than that induced by the rol ABC genes together (75-fold increase). In contrast, the rol C gene alone was as efficient as the rol ABC genes together (mean value of total alkaloids: 0.36% dry weight, i.e., 12-fold times more than in untransformed roots) to stimulate the biosynthesis of tropane alkaloids in A. belladonna hairy root cultures.

Atropa belladonna↗
PubMed1999

An Atropa belladonna hyoscyamine 6beta-hydroxylase gene is differentially expressed in the root pericycle and anthers.

The AbH6H gene for hyoscyamine 6beta-hydroxylase (H6H), which converts hyoscyamine to scopolamine, was isolated from Atropa belladonna. This plant also possesses a related sequence, Ab psiH6H, which appears to be a non-functional pseudo-gene. AbH6H RNA was detected in cultured root, native root and anther, but not in stem, leaf, pistil, petal, and sepal tissues. In situ hybridization, immunohistochemistry and promoter::GUS transgene analysis showed that AbH6H is expressed specifically in root pericycle cells, and in tapetum and pollen mother cells. A 671 bp 5'-upstream region from AbH6H was sufficient for pericycle-specific expression in hairy roots of A. belladonna and Hyoscyamus niger, which both produce scopolamine, but cell-specific regulation was severely compromised in tobacco hairy roots, which do not produce scopolamine.

Amino Acid Sequence↗
PubMed2006

Tracking ancient polyploids: a retroposon insertion reveals an extinct diploid ancestor in the polyploid origin of belladonna.

Polyploidy is a prominent process in plant evolution and adaptation, but molecular phylogenetic studies of polyploids based on DNA sequences have often been confounded by their complex gene and genome histories. We report here a retroposon insertion in the nuclear gene granule-bound starch synthase I (GBSSI or "waxy") that clearly reveals the ancient hybrid history of the medically important polyploid species belladonna (Atropa belladonna) and resolves the controversy over the taxonomic group to which it belongs, the tribe Hyoscyameae (Solanaceae). Our inferences based on the pattern of presence or absence of the retroposon insertion are corroborated by phylogenetic analyses of the GBSSI gene sequences. This case may suggest that retroposons are promising molecular markers to study polyploid evolution.

Atropa belladonna↗
PubMed1997

[Atropa belladonna poisoning suggesting severe post-traumatic brain damage].

In the paper I have described a case of a student of the University of Technology who has suffered a severe Atropa belladonna intoxication. Originally the student has been sent to the Surgery Department by the ambulance service doctor who had recognized the symptoms of the post-traumatic brain damage. The diagnosis of the post-traumatic brain damage has been excluded during the following diagnostic investigation, the symptoms represented by the patient and his bad general condition, however, has still reminded the same. As the next step the patient has been transferred to the Toxicology Ward according to our suggestion of Atropa belladonna intoxication, where the diagnosis has been confirmed. The patient has regained consciousness after 10 hours of the therapy (with the use of the respirator) and has been discharged from the hospital in a good general condition after 11 days. This case of intoxication has been an example of an experiment done by a very inquiring student on himself.

Adult↗
PubMed2000

Learning deficits induced by 4 belladonna alkaloids are preferentially attenuated by tacrine.

AIM: To examine the antagonism of tacrine on the amnesic effects of scopolamine (Sco), anisodine (AT3), atropine (Atr), and anisodamine (Ani). METHODS: Cognitive functions and locomotor activities were determined using two sessions of step-through and open-field tests, respectively. Mice were injected with one of the belladonna alkaloids (0.05-50 mumol.kg-1, i.p.) and tacrine (50 mumol.kg-1, s.c.) 30 min before the first session. RESULTS: Tacrine completely blocked the avoidance-learning deficit caused by Sco 0.5 mumol.kg-1, AT3 and Atr 5 mumol.kg-1, or Ani 50 mumol.kg-1. But tacrine partly antagonized the learning deficit induced by Sco 5-50 mumol.kg-1 or Atr and AT3 50 mumol.kg-1. The avoidance-memory deficit caused by Sco 0.05-5 mumol.kg-1 or Atr 5 mumol.kg-1 was completely or partly attenuated by tacrine, which did not antagonize the memory deficit elicited by Sco and Atr 50 mumol.kg-1, AT3 5 and 50 mumol.kg-1, and Ani 50 mumol.kg-1. During the acquisition, the locomotor activity of the mice was inhibited by tacrine. This reduction was completely antagonized by Sco 0.5-50 mumol.kg-1, AT3 5-50 mumol.kg-1, Atr 5-50 mumol.kg-1, and only partly antagonized by AT3 and Atr 0.5 mumol.kg-1 or Ani 50 mumol.kg-1. CONCLUSION: Compared with the avoidance-memory deficit, the avoidance-learning deficit caused by belladonna alkaloids is more preferentially attenuated by tacrine.

Animals↗
PubMed2001

The effect of nitrate and ammonium concentrations on growth and alkaloid accumulation of Atropa belladonna hairy roots.

The growth, the alkaloid production, as well as the scopolamine/hyoscyamine ratio of two clones of belladonna hairy roots were studied. The effects of nitrate and ammonium concentrations on these cultures were investigated. A rise in ammonium concentration caused the decline of the hairy roots, while nitrate had a marked effect on the alkaloid content. The alkaloid production obtained with 15.8 mM of NO3- and 20.5 mM of NH4+ was 1.2-1.4 times higher than that obtained when the roots were grown in the standard Murashige and Skoog medium (MS medium, 39.5 mM of NO3- and 20.5 mM of NH4+). The nitrate and ammonium concentrations in the culture medium also had a strong influence on the scopolamine/hyoscyamine ratio. When nitrate or ammonium concentrations were raised, that ratio also was increased 2-3-fold. The hairy root clones originating from transformations with two distinct strains of Agrobacterium had similar responses.

Atropa belladonna↗
PubMed2001

Bimodal dose-dependent effect on autonomic, cardiac control after oral administration of Atropa belladonna.

This single-blind placebo-controlled study was designed to investigate the dose-dependent vagolytic and vagotonic effects after a single oral administration of Atropa belladonna tincture (ABT, 0.1 mg/ml alkaloid concentration, atropine/scopolamine = 20:1). In eight healthy young subjects, heart rate and noninvasive arterial finger blood pressure were recorded simultaneously over 4 h after oral application of four different doses of ABT (day 1: 2 ml, day 2: placebo, day 3: 5 ml, day 4: 1 ml). On each day, 14 20-min sequences under controlled experimental conditions were performed. Among others, mean RR interval (RR), high-frequency spectral power of heart rate variability (HF), and noninvasive baroreflex sensitivity (BRS) were calculated during metronome breathing in supine position. These parameters were robust markers of vagal activity. One hour after 5ml ABT, RR, HF and BRS decreased clearly in six of eight subjects. This effect was interpreted as vagolytic response. After 1 and 2 ml ABT, and after placebo, RR and HF increased markedly. The increase after ABT was much higher than the increase solely due to adaptation after placebo administration, and it could be clearly identified as an augmentation of vagal cardiac activity caused by low-dose ABT. In conclusion, low doses of orally administered ABT can be effectively used to stimulate parasympathetic activity in man. The mode of vagal activation changes between 2 and 5 ml ABT from vagotonic to vagolytic. ABT has no or very little effect on blood pressure control.

Administration, Oral↗
PubMed1975

Quantitative determination of alkaloids in belladonna tincture USP.

A simplified method for the quantitative analysis of hyoscyamine hydrobromide or atropine in Belladonna Tincture USP is described. The procedure is based on the formation of an alkaloid-dye complex which can be extracted with an organic solvent and measured spectrophotometrically. The results obtained by this method compare favorably with those obtained by the USP method.

Atropa belladonna↗