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The complex-type oligosaccharide binding lectin Datura stramonium agglutinin detects type II A muscle fibres in the branchial biceps from man and cat.

Complex-type oligosaccharides were detected in the sarcoplasm of muscle fibres from cat and human biceps using lectins and anticarbohydrate antibodies. The lectin Datura stramonium agglutinin strongly stained type II A fibres as identified by myosin ATPase activity after alkaline and acid preincubation. In contrast, all muscle fibres showed a moderate coarse granular staining after incubation with Tetracarpidum conophorum agglutinin and Telfairia occidentalis agglutinin which recognize tri-antennary complex glycans poorly bound by D. stramonium agglutinin. Strong sarcoplasmic staining in all muscle fibres was obtained after incubation with an antibody against branched N-acetyllactosamine structure while an antibody against binary 2 --> 3 sialyllactosamine glycans failed to detect the muscle fibres. Treatment of the muscle sections with sialidase prior to incubation with D. stramonium agglutinin did not influence the lectin staining pattern. Staining of blots from electrophoretically separated muscle proteins obtained by homogenization, solubilization and centrifugation of small muscle pieces showed D. stramonium agglutinin binding to a number of bands ranging from 200 kDa to 30 kDa. No D. stramonium agglutinin positive bands were observed in blots from separated mitochondrial proteins while blots from sarcoplasmic reticulum separated by electrophoresis stained many bands in the range from 200 kDa to 30 kDa. It may be concluded that all muscle fibres in human and cat biceps hold intracellular non-sialylated complex-type oligosaccharides and further, that a specific tri-antennary complex-type glycoform is strongly expressed in type II A fibres as recognized by D. stramonium agglutinin. These results indicate a different glycosylation of certain myofibrillar-associated proteins in muscle fibre types.

Animals↗

Protein chemotaxonomy of genus Datura: identical amino acid sequence of ferredoxin from two varieties of Datura stramonium.

The complete amino acid sequence of the respective [2Fe-2S] ferredoxins from two varieties of Datura stramonium, D. stramonium var. stramonium and D. stramonium var. tatula, have been determined by automated Edman degradation of the entire Cm-protein and of the peptides obtained by tryptic digestion and CNBr treatment. The ferredoxins from the two plants exhibited identical amino acid sequences, suggesting a very close taxonomic relationship between the two. This result supports the proposal by Blakeslee and others that these plants, at first named by Linnaeus as two distinct species, i.e. D. stramonium L. and D. tatula L., should be considered two varieties of a single species.

Amino Acid Sequence↗

Lectin binding studies on murine peritoneal cells: physicochemical characterization of the binding of lectins from Datura stramonium, Evonymus europaea, and Griffonia simplicifolia to murine peritoneal cells.

Purified 125I-labeled lectins from Datura stramonium, Evonymus europaea, and Griffonia simplicifolia (I-B4 isolectin) were used to analyze changes in the expression of carbohydrates on the surface of resident (PC) and thioglycollate-stimulated murine (C57B/6J) peritoneal exudate cells (PEC). The lectins from D. stramonium, E. europaea, and G. simplicifolia I-B4 bind specifically to PEC with relatively high affinity (Kd = 5.65 +/- 1.08 X 10(-7) M, 1.08 +/- 0.12 X 10(-8) M, and 1.33 +/- 0.15 X 10(-7) M, respectively). Assuming a single lectin molecule binds to each cell surface saccharide, the number of receptor sites per cell ranged for different cell samples from 22.3 to 50.0 X 10(6), from 3.8 to 4.8 X 10(6), and from 2.0 to 16.8 X 10(6) for D. stramonium, E. europaea, and G. simplicifolia I-B4 lectins, respectively. There were approximately 3- to 7-fold, 16- to 20-fold, and 2- to 20-fold increases in binding capacity for D. stramonium, E. europaea and G. simplicifolia I-B4, respectively, compared to the binding to resident, peritoneal cells. Scatchard plots of the binding of all three lectins to PEC were linear, suggesting that the receptor sites for these lectins are homogeneous and noninteracting. The binding capacity of these lectins to PEC was unchanged after trypsin digestion of cells. The expression of carbohydrates on the surface of PEC was also monitored by an agglutination assay. PEC were agglutinated by all three lectins whereas PC either were not agglutinated or were agglutinated only at high lectin concentrations. On the basis of our knowledge of the carbohydrate binding specificity of the D. stramonium and G. simplicifolia I-B4 lectins, we postulate that, parallel with thioglycolate stimulation, there is an increase in the number of N-acetyllactosamine residues and terminal alpha-D-galactosyl end groups. The blood group B, and H type 1 determinants--DGa1 alpha 1,3[LFuc alpha 1,2]DGa1 beta 1,3(or 4)DGlcNAc and LFuc alpha 1,2DGa1 beta 1,3DG1cNAc, respectively, as well as DGa1 alpha 1,3DGa1 beta 1,3(or 4)DGlcNAc--may be considered to be possible receptors for the E. europaea lectin. These glycoconjugates, present on the surface of peritoneal exudate cells, provide new chemical markers for studying the differentiation of resident peritoneal cells.

Agglutination Tests↗

Alkaloid patterns in some varieties of Datura stramonium.

A comparative GC-MS investigation of the alkaloid patterns of three varieties of Datura stramonium vars. stramonium, tatula and godronii, was carried out. Twenty-five tropane alkaloids were identified in the plant organs. Alkaloid patterns of the roots, leaves and seeds of the varieties grown at equal conditions in Bulgaria were very similar. In contrast, alkaloid pattern of D. stramonium var. stramonium, grown in Egypt, showed significant differences indicating that it is influenced more strongly by the environmental factors than genetic ones.

Datura stramonium↗

Inhibition of proliferation and induction of differentiation of glioma cells with Datura stramonium agglutinin.

We found that a lectin, Datura stramonium agglutinin, induced irreversible differentiation in C6 glioma cells. The differentiated cells had long processes, a low rate of proliferation and a high content of glial fibrillary acidic protein. When the medium was replaced with Datura stramonium agglutinin-free medium after 1 h, cell proliferation continued to be inhibited. Experiments with several other lectins indicated that both recognition of linear N-acetyllactosamine repeats and recognition of multiantennary units of cell-surface glycans were required for the inhibition of C6 proliferation. Proliferation of four human glial tumour cells was also inhibited by Datura stramonium agglutinin. Further, these differentiated human glial tumour cells had long processes and a high content of glial fibrillary acidic protein similar to differentiated C6 glioma cells. Taken together, these observations suggest that Datura stramonium agglutinin may be useful as a new therapy for treating glioma without side effects.

Agglutinins↗

[Self-poisoning with Datura stramonium. 3 case reports].

BACKGROUND: Datura stramonium is a hallucinogenic plant that causes serious poisoning. Due to its easy availability and strong anticholinergic properties, substance users and teens may use Datura stramonium as a drug. Consumption of any part of the plant can result in severe toxicity. CASE REPORTS: 3 cases of acute self-poisoning by ingestion of Datura stramonium are reported. The patients presented with a typical anticholinergic syndrome: agitation, confusion, hallucinations and combative behaviour; all of them had mydriasis, but dry mouth and tachycardia were less common. All these 3 subjects had a good prognosis but have required hospitalisation because of severe psychiatric derangement with agitated behaviour. The patients were favourably managed with only symptomatic treatment. DISCUSSION: This article reviews the clinical syndrome associated with the toxicity. The severity of hallucinations and confusion, associated with pupillary dilation, flushing, dry mouth, and tachycardia, are related with Datura intoxication. Symptomatic treatment is efficient. CONCLUSION: Primary care physicians might be informed about the abuse of Datura stramonium, often associated with substance misuse, and the need to educate risk-patients.

Adult↗

[The production of gastrodin through biotransformation of p-hydroxybenzaldehyde by cell suspension culture of Datura stramonium].

AIM: To investigate the production of p-hydroxymethylphenol-beta-D-glucoside (gastrodin) through biotransformation by plant cell suspension cultures. METHODS: Using cell suspension cultures of Datura stramonium to convert the exogenous p-hydroxybenzaldehyde into gastrodin was conducted and the converted compounds were separated with a combination of multi-chromatography. Their chemical structures were determined on the basis of spectral analysis and chemical evidence. RESULTS: The conversion procedure of p-hydroxybenzaldehyde into gastrodin by Datura stramonium cell suspension cultures was established. The synthesized gastrodin (II) was isolated from the fermental liquor and identified by spectral analysis. At the same time, the p-hydroxybenzyl alcohol (I) converted through biotransformation of p-hydroxybenzaldehyde by cell suspension cultures of Datura stramonium was also isolated and identified. Two compounds were also isolated from the cell cultures and they were identified as beta-D-furanoallulose (III) and n-butyloxystyryl-beta-D-pyranoallulose (IV). CONCLUSION: Datura stramonium grown in suspension cultures can convert exogenous p-hydroxybenzaldehyde into the corresponding gastrodin.

Benzaldehydes↗

Metabolism of the herbicide glufosinate-ammonium in plant cell cultures of transgenic (rhizomania-resistant) and non-transgenic sugarbeet (Beta vulgaris), carrot (Daucus carota), purple foxglove (Digitalis purpurea) and thorn apple (Datura stramonium).

The metabolism of the herbicide glufosinate-ammonium was investigated in heterotrophic cell suspension and callus cultures of transgenic (bar-gene) and non-transgenic sugarbeet (Beta vulgaris). Similar studies were performed with suspensions of carrot (Daucus carota), purple foxglove (Digitalis purpurea) and thorn apple (Datura stramonium). 14C-labelled chemicals were the (racemic) glufosinate, L-glufosinate, and D-glufosinate, as well as the metabolites N-acetyl L-glufosinate and 3-(hydroxymethylphosphinyl)propionic acid (MPP). Cellular absorption was generally low, but depended noticeably on plant species, substance and enantiomer. Portions of non-extractable residues ranged from 0.1% to 1.2% of applied 14C. Amounts of soluble metabolites resulting from glufosinate or L-glufosinate were between 0.0% and 26.7% of absorbed 14C in non-transgenic cultures and 28.2% and 59.9% in transgenic sugarbeet. D-Glufosinate, MPP and N-acetyl L-glufosinate proved to be stable. The main metabolite in transgenic sugarbeet was N-acetyl L-glufosinate, besides traces of MPP and 4-(hydroxymethylphosphinyl)butanoic acid (MPB). In non-transgenic sugarbeet, glufosinate was transformed to a limited extent to MPP and trace amounts of MPB. In carrot, D stramonium and D purpurea, MPP was also the main product; MPB was identified as a further trace metabolite in D stramonium and D purpurea.

Aminobutyrates↗

Datura stramonium Agglutinin : Location in the Seed and Release upon Imbibition.

The distribution of Datura stramonium agglutinin over different tissues of D. stramonium L. seeds was visualized by immunocytochemical techniques and quantified by agglutination assays. The lectin occurs predominantly in the outer seed tissues (seed coat and seed epidermis), where it is associated, at least in part, with the cell walls. Developing D. stramonium seeds secrete newly synthesized lectin polypeptides into the incubation medium, which confirms the extracellular location of the lectin. Imbibition of mature decoated seeds results in a rapid and highly specific release of lectin. Indeed, imbibition solutions contain almost exclusively the lectin together with a few other carbohydrate-binding proteins; this is indicative of the predominance of these proteins in the seed surface layer. The presence of important amounts of lectin in the outer tissues of the seed is consistent with a possible role in the mediation of cell-cell interactions.

Journal Article↗

The formation of 3 alpha- and 3 beta-acetoxytropanes by Datura stramonium transformed root cultures involves two acetyl-CoA-dependent acyltransferases.

Tropine (tropan-3 alpha-ol) is an intermediate in the formation of hyoscyamine. An acyltransferase activity that can acetylate tropine using acetylcoenzyme A as cosubstrate has been found in transformed root cultures of Datura stramonium. A further acyltransferase activity that acetylates pseudotropine (tropan-3 beta-ol) with acetyl-coenzyme A is also present. These two activities can be partially resolved by anion-exchange chromatography, some fractions containing only the pseudotropine-utilizing activity. The basic properties of these two enzymes are reported and their roles in forming the observed alkaloid spectrum of D. stramonium roots discussed.

Acetyltransferases↗

Altered nitrogen metabolism associated with de-differentiated suspension cultures derived from root cultures of Datura stramonium studied by heteronuclear multiple bond coherence (HMBC) NMR spectroscopy.

De-differentiation of transformed root cultures of Datura stramonium has previously been shown to cause a loss of tropane alkaloid synthetic capacity. This indicates a marked shift in physiological status, notably in the flux of primary metabolites into tropane alkaloids. Nitrogen metabolism in transformed root cultures of D. stramonium (an alkaloid-producing system) and de-differentiated suspension cultures derived therefrom (a non-producing system) has been compared using Nuclear Magnetic Resonance (NMR) spectroscopy. (15)N-Labelled precursors [((15)NH(4))(2)SO(4) and K(15)NO(3)] were fed and their incorporation into nitrogenous metabolites studied using Heteronuclear Multiple Bond Coherence (HMBC) NMR spectroscopy. In both cultures, the same amino acids were resolved in the HMBC spectra. However, marked differences were found in the intensity of labelling of a range of nitrogenous compounds. In differentiated root cultures, cross-peaks corresponding to secondary metabolites, such as tropine, were observed, whereas these were absent in the de-differentiated cultures. By contrast, N- acetylputrescine and gamma-aminobutyric acid (GABA) accumulated in the de-differentiated cultures to a much larger extent than in the root cultures. It can therefore be suggested that the loss of alkaloid biosynthesis was compensated by the diversion of putrescine metabolism away from the tropane pathway and toward the synthesis of GABA via N-acetylputrescine.

Alkaloids↗

GC-MS investigation of tropane alkaloids in Datura stramonium.

Alkaloids, GS-MS, Datura stramonium The alkaloid spectrum in roots, leaves and seeds of Datura stramonium L. was investigated by GC-MS. Twenty-nine tropane alkaloids are detected. Twelve of them are new constituents for the species and the two tropane esters 3-(3'-acetoxytropoyloxy)tropane (21) and 3-(2'-hydroxytropoyloxy)tropane (26) are described for the first time.

Alkaloids↗

[A case of datura stramonium poisoning--general problems of differential diagnosis].

In past year drug abuse becomes more and more general in Hungary. In addition to consume traditional drugs, other substances are used frequently too. One of them is the Datura stramonium, which contains alkaloids (mostly atropine), and can result in hallucinations. Therefore Datura stramonium is seemingly becoming popular as a hallucinogenic drug. The consumption of any part of the plant causes atropine intoxication, thus anticholinergic delirium. Differential diagnosis of Datura intoxication can be difficult in the everyday medical practise. In our paper the symptomatology, diagnosis, differential diagnosis, and therapy of Datura intoxication are discussed and we report one of our cases.

Adult↗

Accidental Datura stramonium poisoning in a dog.

Datura stramonium is potentially poisonous to humans and livestock; however, there's little description of clinical and pathological findings in dogs naturally intoxicated. We report an accidental Datura stramonium poisoning in a dog emphasizing the importance of recognizing the classical signs of anticholinergic poisoning.

Animals↗

[Voluntary poisoning by ingestion of Datura stramonium. Another cause of hospitalization in youth seeking strong sensations].

Natural hallucinogenic substances have been used in numerous cultures throughout the world for millenniums, providing a sacred intermediary between men and gods. They were used during initiatory, divinatory rites and played an essential role in the myths of exorcism and charm where the initiated enters a sort of trance which separates the soul from the body before "revival". It resulted that a number of plants were given names reflecting their relationship with the divinities, bestowing upon them an aura of mystery and sacredness. With the development of free information superhighways via the net, recipes using these hallucinogenic substances can now travel anywhere on the planet, offering the non-initiated a means of experimenting their search for strong sensations. At the same time, the science of botany has undergone a major revolution. Patient observation of nature is no longer necessary since a well-conducted computer search can easily yield a large supply of plants offered for on-line purchase. Children and adolescents in primary and secondary schools are becoming a new target for the botanical market: young people exchange the latest recipes and Internet addresses, or more simply the objects of their new on-line purchase. This has led to an outbreak in the use of mushrooms and hallucinogenic plants where the cultural notion of initiation or sacred rites has metamorphosed into a phenomenon of communication between young people, notably during festive parties. Datura stramonium has become quite popular and the number of hospitalizations for acute datura poisoning has increased rapidly, as indicated by the statistics of the Bordeaux Poison Center (14 hospitalizations for voluntary poisonings with datura in 2002, compared with 5 in 2001). Besides the self-inflicted risk, one of the most important problems is the delay to diagnosis, simply because the primary care physician did not think of such an "exotic" cause. We report here the case of a 22-year-old young man hospitalized following ingestion Datura stramonium seeds. He presented confusions and dreamlike frenzy. The clinical course was favorable in 72 hours with symptomatic treatment, marked by amnesia of the episode.

Adult↗