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PubMed1988

Effect of glycosylation inhibitors on the binding of insulin and lectins and on hormonal imprinting in Tetrahymena.

Treatment of the unicellular Tetrahymena with inhibitors of glycosylation (glucosamine or 2-fluoro-2-deoxy-D-glucose) leads, with a few exceptions, to enhancement of the binding of FITC-labelled insulin and lectins (Con-A, Helix, Datura). The development of insulin-evoked imprinting (i.e., the enhancement of binding at subsequent contacts) is not prevented by the administration of glucosamine. Meanwhile the administration of fluoroglucose results, on its own, in enhanced binding. This increased binding capacity cannot be further enhanced by imprinting.

Animals↗

PubMedDate not supplied

[Mannan sulfates--inducers of plant resistance to viral infection].

Mannan sulphates (MS) synthesized on the basis of extracellular linear mannan (LM) of Rhodotorula rubra induce resistance of Immune-580 tobacco and thornapple (Datura stramonium) to tobacco mosaic virus (TMV). The resistance is manifested in a decrease in the number and/or size of viral local lesions (LL) in MS-treated (mg/ml) leaf halves. The reduction in the LL size does not seem to be due to the direct inhibition of TMV multiplication by the polysaccharide as the virus accumulation in the tissues of the systemic host (Samsun tobacco) does not decrease when MS is used 15 min after infection. The induced resistance is partially inhibited by actinomycin D and completely by heating (32 degrees C). Unlike MS, neutral LM decreases only the number of LL on the treated parts of the tissue but does not influence their size. The antiviral activity of LM does not change in the presence of actinomycin D. The possible mechanisms of the LM and MS protective effect in plants are discussed.

Dactinomycin↗

PubMed1978

North American Indian rock art and hallucinogenic drugs.

It is proposed that the aboriginal rock paintings in two areas of North America may have been produced by shamans while they were under the influence of hallucinogenic agents derived from plants. The first of these areas is the Chumash and Yokuts Indian region of California, where polychrome paintings show designs similar to those visualized during the trance induced by decoctions of jimsonweed (Datura species). The second area is the lower Pecos River region of Texas, where shamanistic figures display traits considered to be conceptual analogues of the mescal bean (Sophora secundiflora) cult as practiced during historic times by Great Plains Indians. Although the evidence is only circumstantial, the proposed connections between these rock drawings and mind-expanding pharmacologic compounds fit well into the documented relationship that encompasses hallucinogenic drugs and certain movable objects of pre-Columbian American art.

Art↗

PubMed1983

Effects of herbicides on the concentration of poisonous compounds in plants: a review.

Herbicides may raise, lower, or not affect the concentration of poisonous compounds in plants. Herbicides, 2,4,5-T and silvex controlled Wasatch milkvetch (Astragalus miser var oblongifolius) and markedly reduced the concentration of miserotoxin, a poisonous aliphatic nitro compound. The concentration of phototoxic furocoumarins in the leaves of spring parsley (Cymopterus watsonii) decreased to nontoxic concentrations 4 to 5 weeks after treatment with 2,4-D. The hydrocyanic acid content of wild cherry (Prunus spp) decreased after treatment with 2,4-D and 2,4,5-T, but increased in Sudangrass (Sorghum halapense) after treatment with 2,4-D. The alkaloid concentration in Barbey larkspur (Delphinium barbeyi) increased after treatment with 2,4,5-T or silvex, decreased in horsetail (Equisetum palustre) after treatment with MCPA, and was unaffected in Datura after treatment with several herbicides. The effect of herbicides on nitrate concentration in plants was dependent upon the species of plant and the herbicide used.

Alkaloids↗

PubMed1984

Expression of disease symptoms in cauliflower mosaic virus genomic hybrids.

In an effort to determine if particular regions of the cauliflower mosaic virus (CaMV) genome could be associated with particular phenotypic characters, strains of CaMV differing markedly in biological properties were recombined to produce hybrids. DNA from pairs of (infectious) genomic clones was cleaved with restriction endonucleases, then mixed and ligated. Recombinants were found by screening transformants in E. coli, or by selection in vivo for infectious hybrids. Recombinants in infected turnip plants were characterized by restriction endonuclease mapping of their DNA to confirm the hybrid genotype. New hybrid strains that induced less severe disease, or conversely, more severe disease than either parent were observed. The experiments revealed that typical disease expression, consisting of leaf chlorosis and mottling, mapped to a genome segment containing open reading frame VI (ORF VI) and the full-length promoter. This basic disease symptom was found to be influenced by other regions of the genome. Insect transmissibility mapped to ORF II. The ability to develop generalized infections in solanaceous plants was tested in hybrids between CaMV CM1841 and a variant that infects Datura stramonium systemically. In this case the systemic mobilization of virus appeared to be controlled by ORF VI, suggesting that this gene may function in cell-to-cell movement of virus.

DNA, Recombinant↗

PubMed1984

The distribution of repeating [Gal beta 1,4GlcNAc beta 1,3] sequences in asparagine-linked oligosaccharides of the mouse lymphoma cell lines BW5147 and PHAR 2.1.

The occurrence and distribution of the repeating disaccharide [Gal beta 1,4GlcNAc beta 1,3] in the different types of Asn-linked oligosaccharides in mouse lymphoma BW5147 cells have been studied. Glycopeptides were prepared from cells grown in medium containing [6-3H]galactose, and the bi-, tri-, and tetraantennary Asn-linked oligosaccharides were fractionated by serial lectin affinity chromatography on concanavalin A-Sepharose, pea lectin -Sepharose, leukoagglutinating phytohemagglutinin-agarose, and Datura stramonium agglutinin-agarose. As described in this report, the latter lectin binds glycopeptides that contain either the repeating N-acetyllactosamine sequence or an outer mannose residue substituted at C-2 and C-6 by N-acetyllactosamine. The isolated glycopeptides were subjected to methylation analysis, specific exoglycosidase treatments, and digestion with Escherichia freundii endo-beta-galactosidase. Our data indicate that approximately two-thirds of the tetraantennary and one-half of the triantennary Asn-linked oligosaccharides contain repeating N-acetyllactosamine sequences in at least one branch. Many of the repeating sequences contain an additional galactose residue linked alpha 1,3 to a penultimate galactose residue. By contrast, less than 10% of the biantennary oligosaccharides contain the repeating disaccharide. The distribution of the repeating N-acetyllactosamine unit was also examined in a cell line ( PHAR 2.1) that is deficient in UDP-GlcNAc:alpha-mannoside beta 1,6-N-acetylglucosaminyltransferase. These cells are unable to synthesize tetraantennary and certain triantennary species and instead accumulate biantennary oligosaccharides. The total content of repeating N-acetyllactosamine units is greatly decreased in this line, and those that are present are found predominantly in triantennary Asn-linked oligosaccharides. These results demonstrate that the repeating N-acetyllactosamine sequence occurs commonly in complex-type Asn-linked oligosaccharides in BW5147 cells but is confined primarily to tri- and teraantennary species.

Acetylglucosamine↗

PubMed1981

[Preliminary study of guacetisal in chronic bronchitis].

A single-blind, between-patients trial was run on two randomly formed groups of 15 hospitalised male patients with chronic obstructive bronchitis, carefully selected to rule out excessive interference on the part of other pathologies or lund complications, in which 3 tablespoonfuls a day Broncaspin suspension were compared with the same dose of a recognised commercial preparation composed by sodium thymosulphonate, ammonium acetate, codeine, and tincture of datura. The clinical and instrumental data showed that Broncaspin had greater therapeutic activity. The local and general tolerability of the two preparations was uneceptionable.

Aged↗

PubMed1981

Absence of arthrogryposis in newborn Hampshire pigs from sows ingesting toxic levels of jimsonweed during gestation.

No congenital arthrogryposis or other deformities were found in newborn Hampshire pigs born to sows administered Datura stramonium (jimsonweed) during the second or third months of pregnancy at levels producing marked clinical signs of toxicity in the dams. The results suggest that jimsonweed is not teratogenic in pigs, contrary to epidemiologic speculation by other workers.

Animals↗

PubMed1995

Evaluation with receptor binding assay on the water extracts of ten CNS-active Chinese herbal drugs.

In the present study, we established receptor binding assays to evaluate the water extracts of ten central nervous system (CNS)-active Chinese herbal drugs. These ten herbal drugs are Chaihu (Radix Bupleuri), Chuanxiong (Rhizoma Chuanxiong), Danggui (Radix Angelicae sinensis), Danshen (Radix Salviae miltiorrhizae), Duhuo (Radix Angelicae pubescentis), Hangqin (Radix Scutellariae), Qinjiao (Radix Gentianae macrophyllae), Shengma (Rhizoma Cimicifugae), Suanzaoren (Semen Zizphi spinose), and Yangjihua (Flos Daturae). The results indicated that these water extracts contained the principles acting on the dopamine (D1 & D2), muscarinice acetylcholine (M1), or 5-HT (5-HT1A & 5-HT2) receptors, or the benzodiazepine and the gamma-amino-n-butyric acid (GABA) binding sites of GABAA receptors as determined by receptor binding assays. The receptors or binding sites which predominantly acted by each water extract are listed as follows: Chaihu: D2, 5-HT1A, GABA; Chuanxiong: GABA, 5-HT1A; Danggui: GABA, 5-HT1A; Danshen: BDZ; Duhuo: GABA, 5-HT1A, D2, D1; Hangqin: BDZ, D1, 5-HT1A; Qinjiao: GABA, BDZ, 5-HT1A, D2; Shengma: 5-HT1A; Suanzaoren: 5-HT1A, 5-HT2, GABA; Yangjihua: M1, 5-HT1A, 5-HT2. These results provided evidence to explain the CNS effects of these herbal drugs at the receptor level. Furthermore, these results provided information to direct the isolation and purification of receptor-interactive compounds from these herbal drugs.

Animals↗

PubMed1995

[The effect of steroids on organ-cultured porcine trabecular meshwork--an ultrastructural, biochemical, and lectin histochemical study].

The effect of corticosteroid administration on porcine trabecular meshwork (PTM) was studied by ultrastructural and biochemical analysis. Electron microscopic observations showed an increase of elastic fibers in PTM treated with steroids. Two-dimensional polyacrylamide gel electrophoresis with silver staining showed an increase of proteins with molecular weights of 52 kDa, 66 kDa and 70 kDa. The 66 kDa protein was assumed to be a glycoprotein containing N-acetyl-D-glucosamine, as it was stained positively in the lectin blot analysis with Triticum vulgaris (WGA) and Datura stramonium (DSA). The elastic fibers reacted positively to WGA and DSA, and the degree of this reaction was strengthened by steroid treatment. These results demonstrated that steroid treatment induces ultrastructural and biochemical changes in PTM, and an increase of elastic fibers containing glycoprotein with N-acetyl-D-glucosamine.

Acetylglucosamine↗

PubMed1994

Cytokeratin peptide SFGSGFGGGY mimics N-acetyl-beta-D-glucosamine in reaction with antibodies and lectins, and induces in vivo anti-carbohydrate antibody response.

Recently, we found that human anti-GlcNAc mAbs reacted with keratin from human skin and recognized specific peptide epitopes of human cytokeratin 14. Our data demonstrate that anti-keratin Ab responses in mice might be driven by Ags bearing terminal O-linked GlcNAc residues. To determine if mouse anti-GlcNAc mAbs recognized peptide epitopes of keratin, several mouse anti-GlcNAc mAbs were reacted with a panel of overlapping synthetic decapeptides of the entire amino acid sequence of human cytokeratin 14 in the ELISA. Results of the ELISA demonstrated that three mAbs, HGAC 54, HGAC 78, and 101.4.1, expressed maximal binding activity to keratin peptide B1 with the amino acid sequence SFGSGFGGGY. In addition, we found that mouse anti-cytokeratin 14 mAb CKB-1 recognized the same peptide and expressed anti-GlcNAc activity as well. Keratin peptide B1 was shown to react not only with anti-GlcNAc mAbs but also with several lectins such as wheat germ agglutinin, Datura stramonium lectin, Lycopersicon esculentum lectin, Solanum tuberosum lectin, and lectin from Wisteria floribunda. Reaction of the lectins with solid-phase keratin peptide B1 was inhibited by soluble GlcNAc and was not inhibited by glucose demonstrating specificity of binding. Using a panel of 24 synthetic keratin B1 peptides, each with a single amino acid substitution, we found that aromatic-aromatic and hydrophobic interactions were the major driving forces in the stabilization of the peptide-protein complexes. Finally, we demonstrated that immunization of BALB/c mice with peptide B1 conjugated to BSA induced an anti-GlcNAc Ab response. Results of our experiments indicated that certain peptides may express functional activity similar to GlcNAc and induce in vivo anti-carbohydrate Ab responses.

Acetylglucosamine↗

PubMed1995

Glycoconjugate expression of chondrocytes and perichondrium during hyaline cartilage development in the rat.

Alterations in the expression of glycoconjugate structures during cartilage development in the chondrocranium, nasal skeleton, Meckel's cartilage, limb buds, vertebral bodies and ribs were investigated comparatively in 13 to 21-d-old rat embryos. The binding patterns of 24 biotinylated lectins were analysed in serial sections and compared with results obtained using histochemical methods. Proteoglycan distribution, assessed by conventional staining procedures, was not associated with lectin binding sites. During early fetal development, hyaluronate concentrations were enhanced in areas of prospective chondrogenesis. With few exceptions, the lectins showed a general increase in intensity of binding to mesenchymal structures. Con A (Canavalia ensiformis), DSL (Datura stramonium), and WGA (Triticum vulgare) displayed a ubiquitous distribution of binding sites. After incubation with LCA (Lens culinaris), PSA (Pisum sativum), STL (Solanum tuberosum), and VAA (Viscum album), characteristic differences in binding intensity between focal areas of developing mesenchyme were seen. DBA (Dolichus biflorus), ECL (Erythrina cristagalli), GSL I (Griffonia simplicifolia), LTA (Lotus tetragonobolus), SJA (Saphora japonica), UEA I (Ulex europaeus) and VVL (Vicia villosa) consistently failed to bind. During chondrogenesis a general reduction of lectin staining was detected. In early stages of development GSL II (Griffonia simplicifolia) was a specific marker of the prechondral blastema in the viscerocranium. PNA (Arachis hypogaea) selectively labelled the prevertebral blastema. In contrast, condensing mesenchyme of limb buds and viscerocranium was not stained. Using RCA (Ricinus communis), it was possible to distinguish chondroblasts from mature cells. All chondrocytes were stained by PSA, PHA-E, PHA-L (Phaseolus vulgaris E and L), and WGA, whereas Con A, LCA, and GSL II detected distinct differences between cartilage with different localisations. Cartilage matrix was constantly negative. Applying GSL II it was possible to distinguish specific segments of the perichondrium. From our results we conclude that especially high mannose oligosaccharides are amplified during development. Terminal sialic acid molecules, branched intralaminar glucose and/or mannose, respectively, internal galactose-(beta 1,4)-N-acetylglucosamine sequences as well as galactose-(beta 1,3)-N-acetylgalactosamine sequences in a preterminal position are diffusely distributed in mesenchymal tissue. In contrast, no evidence for the presence of terminal GlcNAc(beta 1,4)GlcNAc sequences and terminal alpha-fucosyl residues in (1,2) or (1,3)-linkage was obtained. Chondrogenesis appears to be correlated with a general reduction in the extent of expression of oligosaccharide structures. No proof of terminal N-acetylgalactosamine and alpha-galactose moieties was found, whereas our staining results document the expression of terminal beta-galactose structures in restricted areas of the developing mesenchyme.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylglucosamine↗

PubMed1979

The biosynthetic route from ornithine to proline.

It is shown by tracer experiments with DL-[2-3H,5-14C]- and DL-[(RS)-5-3H,5-14C]rnithine, that the metabolic conversion of ornithine into proline, in three plant species (Nicotiana tabacum, Datura stramonium, and Lupinus angustifolius), takes place with maintenance of the delta-hydrogen atoms but with loss of the alpha-hydrogen atom. This indicates a route via alpha-keto-delta-aminovaleric acid (5-amino-2-oxopentanoic acid) and disproves the accepted route via glutamic gamma-semialdehyde (2-amino-5-oxopentanoic acid).

Carbon Radioisotopes↗

PubMed1995

Expression of membrane glycoconjugates on sheep lung endothelium.

BACKGROUND: Carbohydrates play an important role in both the regulation and expression of endothelial cell surface molecules. EXPERIMENTAL DESIGN: To investigate the molecular phenotype of endothelial cells in the sheep lung, we used a panel of 39 lectins and carefully titrated lectin histochemistry to identify small vessel glycoconjugates. Expression of glycoconjugates on isolated cells was studied by flow cytometry and lectin Western blotting. RESULTS: Eight lectins stained small vessel endothelium with minimal background staining. To confirm cell surface binding, endothelial cells from the peripheral lung were isolated and tested for lectin recognition by flow cytometry. Three lectins (Datura stramonium, Griffonia simplicifolia-1, Lycopersicon esculentum) also stained isolated lung cells by flow cytometry. A lectin Western transfer technique demonstrated common binding to a high molecular weight (100 to 130 kD) band. CONCLUSIONS: The use of lectins as probes of cell surface carbohydrate expression supports the possibility of selective glycoconjugate expression on sheep endothelium.

Animals↗

PubMed1995

Mucolipidosis IV: morphology and histochemistry of an autopsy case.

Mucolipidosis Type IV is a rare, autosomal recessive disorder characterized by corneal opacification, mental retardation, and delayed motor milestones. Whereas lysosomal storage material has been demonstrated in biopsied tissues and leukocytes, the complete autopsy pathology, including neuropathology, is unknown. The metabolic defect remains speculative. We report the general and neuropathologic findings of the only known autopsy. In the central nervous system, neuronal loss in the cerebral cortex, basal ganglia, deep cerebellar nuclei, and brainstem nuclei was marked by astrocytosis; the cytoplasm of residual neurons had brown granules. These granules were positive with periodic acid-Schiff, Concanavalia ensiformis, and Sudan black, but not with Luxol-fast blue. Ultrastructurally, neurons contained lysosomes laden with osmiophilic, amorphous and granular material, and few lamellated membrane structures. Hepatocytes, epithelia, endothelia, chondrocytes, and tissue macrophages also stained positively with Datura stramonium and Ricinus communis-I agglutinins, with renal glomeruli also staining with peanut agglutinin; most non-neural cells contained osmiophilic granules on toluidine blue-stained, plastic embedded sections, corresponding to lamellated membrane structures. These findings complement the previously reported ocular morphology and brain and liver biochemistry performed in the same patient, and suggest that the storage material in neurons differs from that in non-neural cells. Furthermore, the underlying defect is not likely to be a deficiency of a single enzyme (i.e. a lysosomal hydrolase).

Adult↗

PubMed1994

Herbal poisoning caused by adulterants or erroneous substitutes.

Six cases of herbal poisoning involving six patients in Hong Kong, Taipei and Kuala Lumpur are reported. The sources of poisoning were identified as adulterants (Podophyllum emodi) or erroneous substitutes (Datura metel). In cases of suspected herbal poisoning, it is recommended that the prescriptions, herbal residues and herb samples should be collected for pharmacognostical and chemical analysis to substantiate the cause of poisoning. Insofar as it is possible, an estimate of the amount of herbs consumed should also be obtained, to establish whether the amount of toxin present is sufficient to account for the symptoms.

Drug Contamination↗

PubMed1994

Characterization of a novel family of ciliary body glycoproteins.

PURPOSE: To isolate and characterize ciliary body epithelial antigens reactive with a monoclonal antibody, 2B4.14.1. METHODS: A mouse monoclonal antibody generated against human corneal endothelium, 2B4.14.1 reacts with nonpigmented epithelium of human and guinea pig ciliary bodies. The ciliary body proteins reactive with 2B4.14.1 were identified by Western blotting and were partially purified by affinity chromatography with 2B4.14.1 coupled to a solid support matrix. Carbohydrate components of the antigenic molecules were analyzed by lectin chromatography and by digestion with the enzymes N-glycosidase F and endoglycosidase F. The cellular and subcellular distribution of the antigens was examined by immunoperoxidase staining and by immunoelectron microscopy of ultracryotome sections of ciliary body. RESULTS: 2B4.14.1 reacted with families of guinea pig and human ciliary body glycoproteins with estimated molecular weights ranging from 250 to 325 kD. In Western blots of samples reduced before electrophoresis, the high molecular weight bands were replaced by weakly reactive bands at 115 to 130 and 210 kD, indicating that the 2B4.14.1 ligands have disulfide bonds. 2B4.14.1 ligands from both guinea pig and human ciliary body were bound by immobilized lectins, including concanavalin A, Datura stramonium lectin, and Lens culinaris hemagglutinin, which recognize components of N-linked oligosaccharides. Guinea pig ciliary body antigens digested with N-glycosidase F and endoglycosidase F failed to react with 2B4.14.1 in Western blots, confirming the presence of N-linked oligosaccharide chains and indicating that they form an integral part of the 2B4.14.1-reactive antigenic site. Molecular weight shifts of glycosidase-digested antigens were consistent with the presence of two to four N-linked oligosaccharide units. In immunoperoxidase-stained sections of guinea pig and human ciliary body, 2B4.14.1 reacted primarily with nonpigmented epithelial cells. Staining of guinea pig epithelial cells was fairly uniform; staining of human epithelial cells was concentrated on the basal surface. By immunoelectron microscopy, a majority of the 2B4.14.1 antigenic reactivity was localized immediately external to the nonpigmented epithelial cell plasma membrane. CONCLUSIONS: 2B4.14.1 reacts with a novel family of high molecular weight glycoproteins associated with the nonpigmented epithelial cell surface in guinea pig and human ciliary body.

Aged↗

PubMed1995

Histochemical demonstration of different types of poly-N-acetyllactosamine structures in human thyroid neoplasms using lectins and endo-beta-galactosidase digestion.

Blood-group-related antigens expressed in papillary carcinomas and other types of neoplasm of the human thyroid glands have been shown to be carried by poly-N-acetyllactosamines containing a linear domain susceptible to endo-beta-galactosidase digestion. To make clear more precisely the backbone poly-N-acetyllactosamine structures, labelled lectins specific to different types of these structures and specific to core structures with beta 1-6GlcNAc branching of N- and O-linked glycoproteins were employed in conjunction with prior endo-beta-galactosidase digestion on formalin-fixed, paraffin-embedded neoplasms of the human thyroid glands. In papillary carcinomas, Datura stramonium agglutinin (DSA) and succinyl wheat germ agglutinin (Suc-WGA) reacted most consistently and frequently with papillary carcinomas from all the individuals examined. Pokeweed mitogen (PWM) likewise stained the cells of papillary carcinomas from all the individuals examined, but in some individuals the number of lectin-reactive cells were very small. Lycoperscion esculentum aggultinin (LEA), Solanum tuberosum agglutinin (STA), Phaseolus vulgaris agglutinin L (PHA-L) and Artocarpus integrifolia agglutinin (jacalin) similarly bound to the cancer cells from most of the individuals, and in these cases the number of reactive cells was usually much more restricted than was the case with DSA or PWM. In adenoma and other types of carcinoma, such as follicular carcinomas, these lectins specific to poly-N-acetyllactosamine exhibited slight or no reactivity with the cells, whereas PHA-L and jacalin similarly bound to the cells of adenomas and carcinomas from most of the individuals examined. Prior digestion with endo-beta-galactosidase completely eliminated or markedly reduced the reactivity with PWM and LEA in papillary carcinomas. Reactivity with DSA, Suc-WGA, STA, PHA-L and jacalin was slightly reduced or not at all affected by enzyme digestion. These results confirmed that poly-N-acetyllactosamine species found in papillary carcinomas are quite different from those in other types of thyroid neoplasm, suggesting that at least three different types of poly-N-acetyllactosamine, that is, linear unbranched short and long sequences and highly branched ones are produced in these cells.

Adult↗