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Large animal hepatotoxic and nephrotoxic plants.

The hepatotoxic and nephrotoxic plants of large domestic animals have been reviewed. The most important ones are those widely distributed as weeds over pastures, negelcted forests and grasslands, those used as ornamentals, the nitrate concentrating forage crops, and the cyanophoric plants. Crotolaria spp, the ragwort (Senecia jacobaea), the lantana spp. and heliotopum are common hepatoxic plants. Amaranthus retroflexus, Datura stramonium, Solanum rostratum, and the castor oil plant (Ricinus communis) are nephrotoxic plants.

Animals↗

Anticholinergic poisoning due to Chinese herbal medicines.

Serious poisoning may occur following the consumption of Chinese herbal medicines (CHM) containing anticholinergics. The great majority of cases are probably related to the use of yangjinhua, the dried flower of Datura metel L, for treating bronchial asthma, chronic bronchitis, pains and flu symptoms. In some cases the use of CHM contaminated by atropine-like substances or fake herbs were suspected. Some ginseng (Panax ginseng) preparations might have been adulterated with Mandragora officinarum (scopolamine) and other herbs.

Cholinergic Antagonists↗

Jimson weed poisoning--Texas, New York, and California, 1994.

Ingestion of Jimson weed (Datura stramonium), which contains the anticholinergics atropine and scopolamine, can cause serious illness or death. Sporadic incidents of intentional misuse have been reported throughout the United States, and clusters of poisonings have occurred among adolescents unaware of its potential adverse effects. This report describes incidents of Jimson weed poisoning that occurred in Texas, New York, and California during June-November 1994.

Adolescent↗

Inhibition of lectin-mediated ovarian tumor cell adhesion by sugar analogs.

Adhesion of A-121 human ovarian carcinoma cells to extracellular matrix is partly mediated via interaction between galaptin, an endogenous beta-galactoside-binding lectin present in extracellular matrix, and specific cell surface carbohydrate receptors identified as lysosomal associated membrane proteins, lamp-1 and lamp-2. In this study, we report that adhesion of human ovarian carcinoma cells to polystyrene plates coated with polymerized human splenic galaptin can be inhibited by polyclonal antibodies raised against lamp-1 and lamp-2 molecules and by pretreatment of A-121 human ovarian carcinoma cells with glucosamine analogs: 2-acetamido-1,4,6-tri-O-acetyl-3- deoxy-3-fluoro-alpha-D-glucopyranose (3-F-GlcNAc) and 2-acetamido-1,3,6-tri-O-acetyl-4-deoxy-4-fluoro-alpha-D-glucopyranose (4-F-GlcNAc). A 48-h exposure of A-121 cells to individual sugar analogs, or to a combination of the two, resulted in a concentration-dependent inhibition of cellular attachment to polymerized galaptin. Both drugs inhibited glycoprotein biosynthesis as measured by cellular incorporation of labeled [3H]glucosamine and [3H]fucose with negligible effects on [3H]thymidine and [3H]leucine incorporation and cell growth. As a result of drug action on glycoprotein biosynthesis, an alteration in the structure of the galaptin receptor was noted by indirect immunofluorescence and Western blot analysis. Moreover, probing gels of cell extracts with anti-lamp antibodies or Datura stramonium lectin demonstrated significant changes in the reactivity and pattern of glycoprotein staining, suggesting an effect of sugar analogs on the glycosylation of various cellular receptor molecules. The greatest change was observed when tumor cells were exposed to a combination of the two sugar analogs. These studies suggest that specific endogenous lectins and their surface receptors play a role in tumor cell adhesion and perhaps metastasis and may serve as suitable targets for therapeutic exploitation.

Acetylglucosamine↗

[Thorn apple poisoning].

A 20-year-old man presented with signs of thornapple intoxication: restlessness, disorientation, hallucinations, euphoria, and furthermore dry and red skin and symmetrical dilation of the pupils. Thornapple intoxication mimics atropine intoxication. Thorn apples (Datura stramonium) are seemingly becoming popular as a hallucinogenic drug. When the symptoms are mild gastric lavage leaving active charcoal in the stomach afterwards will suffice. In case of severe symptoms treatment with physostigmine is indicated.

Adult↗

The dangers of jimson weed and its abuse by teenagers in the Kanawha Valley of West Virginia.

Jimson weed (Datura stramonium, a member of the Belladonna alkyloid family) is a plant growing naturally in West Virginia and has been used as a home remedy since colonial times. Due to its easy availability and strong anticholinergic properties, teens are using Jimson weed as a drug. Plant parts can be brewed as a tea or chewed, and seed pods, commonly known as "pods" or "thorn apples," can be eaten. Side effects from ingesting jimson weed include tachycardia, dry mouth, dilated pupils, blurred vision, hallucinations, confusion, combative behavior, and difficulty urinating. Severe toxicity has been associated with coma and seizures, although death is rare. Treatment consists of activated charcoal and gastric lavage. Esmolol or other beta-blocker may be indicated to reduce severe sinus tachycardia. Seizures, severe hypertension, severe hallucinations, and life-threatening arrhythmias are indicators for the use of the anticholinesterase inhibitor, Physostigmine. This article reviews the cases of nine teenagers who were treated in hospitals in the Kanawha Valley after ingesting jimson weed. We hope this article will help alert primary care physicians about the abuse of jimson weed and inform health officials about the need to educate teens about the dangers of this plant.

Adolescent↗

[Poisoning with Jimson weed. Five cases treated with physostigmine].

During the autumn of 1995, the National Poisons Information Centre was contacted about several cases of poisoning with Jimson weed (Datura stramonium). Five cases are described here. Upon admission to hospital the patients had moderate to severe anticholinergic symptoms, such as mydriasis, sinus tachycardia, agitation, dry mouth, urine retention, fever, hypertension, hallucinations and seizures. Owing to their agitated behaviour, gastrointestinal decontamination was impossible. Repeated doses of physostigmine (2-3 mg) administered intravenously reversed the anticholinergic features without side-effects. In the most severe case, physostigmine was needed for 18 hours (total dose; 25.5 mg). The patients recovered in a day or two, but mydriasis persisted in many cases.

Adolescent↗

Lectin histochemistry of the metathoracic ganglion of the locust Schistocerca gregaria before and after axotomy of the tympanal nerve.

The thoracic ganglia of insects exhibit a highly ordered organization. It seems possible that the information underlying the emergence of this order during development and its maintenance throughout insect life is given via a distinct pattern of molecules distributed within the ganglion. The question we asked was whether the adult insect ganglion is subdivided by the distribution of specific carbohydrates and furthermore whether or not this distribution changes during degeneration and regeneration of neurons. In order to determine the normal carbohydrate distribution, we stained sections of the intact metathoracic ganglion of the locust Schistocerca gregaria with fluorescence-coupled lectins. We succeeded in labeling three sensory neuropil areas with either peanut agglutinin (PNA): Phaseolus vulgaris erythrolectin (PVE), soybean agglutinin, wheat germ agglutinin (WGA), or Vicia villosa agglutinin. Apart from this, PNA, WGA, and succinylated WGA also selectively labeled some neuronal cell bodies, including dorsal unpaired median neurons. Datura stramonium lectin (DSL), Griffonia simplicifolia lectin II, and Solanum tuberosum lectin (STL) bound to glial cells or glia surrounding extracellular matrix. A few lectins stained all structures within the ganglion; some showed no binding at all. In the second part of our study, we tested whether carbohydrates were differentially regulated during transient deafferentation after the axotomy of the tympanal nerve. Binding of PNA and PVE within the auditory neuropil did not change. However, binding of the two glia-associated markers, DSL and STL, clearly differed from that found in intact animals; they bound transiently (day 3-4 until day 10-20 post-surgery) to axonal tracts and neuropils of the axotomized sensory afferents.

Afferent Pathways↗

Analysis of phytochelatin-cadmium complexes from plant tissue culture using nano-electrospray ionization tandem mass spectrometry and capillary liquid chromatography/electrospray ionization tandem mass spectrometry.

Phytochelatins (PCs, also known as class III metallothioneins), a family of sulfhydryl-rich peptides with the formula (gamma-GluCys)(n)Gly(Pc(n), n = 2-11), are induced in plants, yeast and fungi exposed to heavy metals, and are thought to detoxify metals by forming PC- metal complexes. Although PCs have been detected, PC- metal complexes have not been well characterized. In this work, nano-electrospray ionization tandem mass spectrometry (nano-ESI-MS/MS) and capillary liquid chromatography/electrospray ionization tandem mass spectrometry (capillary LC/ESI-MS/MS) methods were used to analyze PC - Cd complexes isolated from Datura innoxia, also known as Jimsonweed, cell culture exposed to Cd. With nano-ESI-MS/MS and capillary LC/ESI-MS/MS we could simultaneously detect the presence of PCs and PC - Cd complexes from plant cell extracts, unambiguously identify these species and elucidate the nature of individual PC - Cd complexes. Phytochelatins with n = 3-6 were detected, as were PC - Cd complexes with PC(3), PC(4) and PC(5). This is the first study to report the size and nature of native PC - Cd complexes from plant tissue samples. These results demonstrate that the direct analysis of plant extracts using nano-ESI-MS/MS and capillary LC/ESI-MS/MS methods is simple and sensitive to the range of PCs and PC - Cd complexes in plants. Hence these methods open up new opportunities for further quantitative analysis of PCs and PC - metal complexes in cell culture and plant systems to understand the relationship between the biosynthesis of these compounds and metal tolerance.

Amino Acid Sequence↗

Structural studies on sugar chains of carbohydrate-deficient transferrin from patients with alcoholic liver disease using lectin affinity electrophoresis.

It is well-known that microheterogeneity of human serum transferrin observed in alcoholics manifests as sialic acid-deficient transferrin isoforms, otherwise known as carbohydrate-deficient transferrin (CDT). A recent study demonstrated that serum CDT lacked one or both of the entire carbohydrate chains but the investigation required several troublesome procedures. The aim of the present study was to confirm the sugar chain structures of serum transferrin, and of serum CDT in particular, from patients with alcoholic liver disease (ALD) using conventional lectin affinity electrophoresis which might be useful in the clinical setting. The serum CDT obtained from ALD-patients was partially purified using an anion exchanger. Serum transferrin and the partially purified serum CDT were investigated by concanavalin A (Con A)- and Datura stramonium agglutinin (DSA)-affinity electrophoresis followed by antibody-affinity blotting and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with Western blotting. By Con A-affinity electrophoresis, serum CDT was separated into weakly reactive and nonreactive transferrins which showed slower electrophoretic mobilities than those from the healthy controls. Moreover, nearly all of the serum CDT was nonreactive with DSA. On SDS-PAGE, the molecular masses of serum CDT were estimated to be approximately 75 and 72 kDa, which corresponded to those of partially and completely deglycosylated transferrin obtained from the healthy controls (78 kDa), respectively. In conclusion, these results indicated that the sugar chain structures of serum CDT from patients with ALD show not merely a loss of terminal sialic acids, but also the absence of asparagine-N-linked oligosaccharides.

Antibody Affinity↗

Capillary electrophoresis-diode array detection--electrospray mass spectrometry for the analysis of selected tropane alkaloids in plant extracts.

Capillary zone electrophoresis, coupled to UV and interfaced with electrospray ionization mass spectrometry (ESI-MS), is described for the simultaneous analysis of hyoscyamine and scopolamine. On-line UV detection occurred at 22 cm from the inlet of the capillary and ESI-MS monitoring was performed along the entire length of the capillary (85 cm). An alkaline solution of 40 mM ammonium acetate at pH 8.5 was suitable for the analysis of the alkaloids under consideration. Under the optimized conditions, including CE and ESI-MS parameters, the two alkaloids were resolved within a short time and with very high sensitivity. The differentiation of hyoscyamine and its positional isomer littorine, commonly encountered in plant material, is also presented using up-front collision-induced dissociation. Finally, the developed method was applied to the analysis of these alkaloids in Belladonna leaf extract and in Datura candida x D. aurea hairy root extract.

Alkaloids↗

Changes of alpha1-acid glycoprotein microheterogeneity in acute inflammation stages analyzed by isoelectric focusing using serum obtained postoperatively.

The relationship between variations of alpha1-acid glycoprotein (orosomucoid, AGP) microheterogeneity detected from isoelectric focusing (IEF) patterns and clinical stage of acute inflammation based on serum C-reactive protein (CRP) levels and interleukin-6 (IL-6) levels was investigated. Serum samples were obtained from healthy subjects, and from patients with esophageal or stomach carcinoma before and after operation. Samples without neuraminidase treatment were used for AGP microheterogeneity analysis, and samples with neuraminidase treatment for AGP heterogeneity analysis. In AGP microheterogeneity, nine bands were detected in the range of pI 3.18-3.57 in sera obtained from healthy subjects. In patients, AGP microheterogeneity changed the first day after operation; the percentage of bands surrounding pI 3.5 increased, and the highest value appeared in sera taken the first or second day after operation and then decreased quickly. These bands showed reactivity for concanavalin A (Con A). The increase in Con A-reactive AGP occurred later than the increase in IL-6, and occurred earlier than the increase in CRP. On the seventh day after operation, the percentage of bands around pI 3.2 increased. These bands showed the reactivity for Datura stramonium agglutinin. On the other hand, in samples with neuraminidase treatment, little change of AGP heterogeneity was observed in most samples, which did not reflect the stage of inflammation. These findings suggested that AGP microheterogeneity detection was a useful marker for the clinical stage of inflammation.

Aged↗

Both cell-surface carbohydrates and protein tyrosine phosphatase are involved in the differentiation of astrocytes in vitro.

Astrocytes are important in the development and maintenance of functions of the CNS, acting in cooperation with neurons and other glial cells. The glycans on astrocyte membrane are believed to play important roles in cell-cell communication. Plant lectins are useful probes, because the lectins can bind to certain cell surface receptors and elicit cellular responses that are normally activated by endogenous ligands for those receptors. In the present study, we investigated the effect of Datura stramonium agglutinin (DSA) on astrocytes and characterized several molecular events. The addition of DSA to a culture of flat, polygonal, immature astrocytes derived from the neonatal rat cerebellum caused the cells to become stellate in shape, similar to astrocytes observed in vivo, concomitant with an increase in expression of astrocyte-specific intermediate filament (glial fibrillary acidic protein [GFAP]) and inhibition of proliferation. These results indicate that DSA binds to astrocytes and triggers differentiation. We also found a decrease in the extent of tyrosine-phosphorylation of a 38-kDa protein. To elucidate the molecular events during astrocyte differentiation, we examined the effects of various signal transduction inhibitors on the transformation from the polygonal to stellate shape (stellation). Interestingly, only tyrosine phosphatase inhibitors, orthovanadate and phenylarsine oxide, showed an inhibitory effect. Our results suggest that DSA induced astrocyte differentiation acts via tyrosine dephosphorylation.

Animals↗

Mucous domains: microchemical heterogeneity in the mucociliary complex of the olfactory epithelium.

Access to and clearance of odorants from binding sites on olfactory cilia are regulated by a complex interplay of molecular, physical and cellular factors. These perireceptor events occur primarily in the mucociliary complex. The use of gold-labelled lectinoprobes, one from Limax flavus (LFA) which is specific for terminal sialic acid residues, and one from Datura stramonium (DSA) specific for N-acetylglucosamine residues, demonstrated intricate patterns of binding in mucous domains of the olfactory mucus and ectodomains of the glycocalyx of olfactory cilia. In electron micrographs of Lowicryl-embedded salamander olfactory mucosa, the mucus consisted of an electron-dense domain that lay superficial to an electron-lucent domain; the interface between the two was irregular. A significantly higher density of binding sites for both lectins was present in the superficial than in the deeper domain. The two domains were not homogeneous: there were small electron-lucent domains (hsL) within the superficial electron-dense domain (hsD) that bound a 4.8-fold lower density of gold-labelled DSA than the surrounding matrix, and the olfactory cilia, which project into hsD, were surrounded by an electron-lucent sheath that appeared to be continuous with the deeper domain. Ectodomains of the glycocalyx associated with olfactory cilia exhibited a higher density of binding sites for both LFA and DSA than did either microvilli of sustentacular cells or respiratory cilia. Specificity of the lectinoprobes was confirmed by inhibition of binding with specific sugars or enzymic removal of specific sugar residues. These results demonstrated microchemical heterogeneity of the non-homogeneous mucous domains in olfactory mucus and in the attendant glycocalyx of olfactory cilia based on the differential localization of sialic acid and N-acetylglucosamine sugar residues.

Animals↗

Dietary phosphorus affects the growth of larval Manduca sexta.

Although phosphorus has long been considered an important factor in the growth of diverse biota such as bacteria, algae, and zooplankton, insect nutrition has classically focused on dietary protein and energy content. However, research in elemental stoichiometry has suggested that primary producer biomass has similar N:P ratios in aquatic and terrestrial systems, and phosphorus-rich herbivores in freshwater systems frequently face phosphorus-limited nutritional conditions. Therefore, herbivorous insects should also be prone to phosphorus limitation. We tested this prediction by rearing Manduca sexta larvae on artificial and natural (Datura wrightii leaves) diets containing varying levels of phosphorus (approximately 0.20, 0.55, or 1.2% phosphorus by dry weight). For both artificial and natural diets, increased dietary phosphorus significantly increased growth rates and body phosphorus contents, and shortened the time to the final instar molt. Caterpillars did not consistently exhibit compensatory feeding for phosphorus on either type of diet. The growth and body phosphorus responses were not explicable by changes in amounts of potassium or calcium, which co-varied with phosphorus in the diets. Concentrations of phosphorus in D. wrightii leaves collected in the field varied over a range in which leaf phosphorus is predicted to affect M. sexta's growth rates. These results suggest that natural variation in dietary phosphorus is likely to affect the growth rate and population dynamics of M. sexta, and perhaps larval insects more generally.

Animals↗

Determination of the enantiomeric purity of scopolamine isolated from plant extract using achiral/chiral coupled column chromatography.

The optical purity of scopolamine derived from Datura sanguinea was determined using coupled column chromatography. A C18 column was used to separate scopolamine from the additional alkaloids and other biological material present in the vegetal extract. The C18 column was coupled through a six-port switching valve to two beta-cyclodextrin columns in series which were used to resolve the scopolamine enantiomers. A single acetylated beta-cyclodextrin column gives equivalent results to the native cyclodextrin columns because of slightly higher enantioselectivity for scopolamine. A multistep extraction procedure is used to isolate scopolamine from the vegetal material. 4-6% of the scopolamine in the final extract was found to be the d enantiomer. Sample extracts as well as commercial scopolamine hydrobromide were treated under various conditions commonly encountered during typical commercial extraction procedures and analyzed in order to determine if the d enantiomer was present in the original material or if it was produced during the extraction process and, if so, determine which step and conditions contribute to racemization. Both the salt and the extract were found to be susceptible to racemization under basic conditions (greater than or equal to pH 9) although the extract appeared to be more susceptible than the salt. Tropic acid formed from the hydrolysis of scopolamine seemed to be completely racemized even though the remaining scopolamine was only partially racemized. Within experimental error, no d enantiomer was found in the original fresh plant material.

Acetylation↗

The macrophage cell surface glycoprotein F4/80 is a highly glycosylated proteoglycan.

Molecules whose expression is limited to particular leukocyte populations are of interest since they may perform unique functions for these cells. We therefore examined the biochemical nature of the F4/80 molecule, which is expressed solely on macrophage and dendritic cell subpopulations. Our study clearly indicates that post-translational modifications, which can influence both a protein's structural and functional features, constitute a major component of the 160-kDa cell-surface F4/80 molecule. The F4/80 molecule is synthesized as a single polypeptide chain which acquires numerous intramolecular disulfide bonds and requires an extended time period (T1/2 = 60 min) for transport to an endoglycosidase H-resistant form. The F4/80 molecule contains extensive N-linked glycosylation which contributes approximately 40 kDa to the mature molecule. The N-linked carbohydrates are of the branched, complex type, containing repeating N-acetylglycosamine or N-acetyllactosamine units which mediate the reactivity of the F4/80 molecule with Datura stramonium lectin. O-linked glycosylation is also present and contributes approximately 10 kDa to the F4/80 molecule. Furthermore, the sialic acid modifications of the F4/80 molecule are primarily through alpha 2-6 linkages to galactose. Finally, we demonstrate that the F4/80 molecule is a proteoglycan modified by chondroitin sulfate glycosaminoglycans. In addition to clarifying the nature of the F4/80 molecule biochemically, these post-translational modifications have specific implications for molecular recognition processes. We conclude that the modifications of the F4/80 molecule may mediate cell-cell recognition, cell adhesion, or ligand binding independently of the F4/80 molecule protein core.

Amino Acid Sequence↗

Glycosylation of beta-1 integrins in B16-F10 mouse melanoma cells as determinant of differential binding and acquisition of biological activity.

Studying B16-F10 cells we could identify beta-1 integrins as laminin, fibronectin and collagen receptors. Gradient ionic strength elution analysis of affinity chromatography showed differential interactions between laminin-binding beta-1 integrins (two beta-1 polypeptides of 105 and 120 kDa) and fibronectin and collagen-binding beta-1 integrins (elution of one major beta-1 polypeptide of 120 kDa) and their respective ligands. To evaluate this diversity we submitted B16-F10 extracts to IEF and SDS-PAGE and found that one beta-1 integrin formed acidic and larger isoforms, while another formed basic and smaller isoforms. To study this difference we also submitted material eluted from WGA-Sepharose columns to IEF but now only the acidic beta-1 isoform was found. Extracts of B16-F10 treated with neuraminidase showed only the basic beta-1 isoform, suggesting that terminal sialic acid residues may be responsible for this acidic pattern, an interpretation supported by the fact that MAA (Maackia ammurensis agglutinin) reacts only with the acidic isoform. Differential glycosylation of beta-1 integrin isoforms in B16-F10 was also demonstrated since the smaller laminin-binding beta-1 integrin isoform reacted only with GNA (Galanthus nivalis agglutinin), whereas the mature larger form reacted with DSA (Datura stramonium agglutinin) and MAA; thus this heterogeneity of beta-1 chains is essentially due to variable glycosylation. Autoradiography and immunoblotting analysis of material separated by 2-dimensional electrophoresis show that only the processed forms of beta-1 integrins are expressed at the cell surface.

Animals↗