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Effect of staurosporine on N-glycosylation and cell adhesion to fibronectin of SW480 human colorectal adenocarcinoma cells.

As N-glycosylation of tumor cell surface proteins affects metastasis of the cells, it was considered that the suppression of metastasis by staurosporine, a protein kinase C inhibitor, is partly caused by changes in N-glycosylation. To examine this possibility, we studied the glycosylation of membrane proteins of SW480 human colorectal adenocarcinoma cells before and after treatment with staurosporine by lectin blot analysis. The results showed that the reactivity of leuko-agglutinating phytohemagglutinin and Datura stramonium agglutinin, both of which bind to highly branched N-linked oligosaccharides characteristic of cancer cells, decreases significantly in the staurosporine-treated cells. In accordance with this, the gene expression of the N-acetylglucosaminyltransferase V, which synthesizes the GlcNAcbeta1-->6 branch of highly branched N-linked oligosaccharides decreased by 30-40% in the drug-treated cells. Since a decrease in the lectin binding was found in several glycoproteins including fibronectin (FN)-receptor, effect of the changes in N-glycosylation of the cells on cell adhesion to FN-matrix was examined. The results showed that the number of cells attached to FN-matrix increases upon treatment of the cells with staurosporine, indicating that the change of N-glycosylation of the FN-receptor promotes cell adhesion to the extracellular matrix, which may lead to the suppression of metastasis of cancer cells.

Adenocarcinoma↗

Xanthine oxidase inhibitory activity of some Indian medical plants.

Xanthine oxidase inhibitory activity was assayed from six species belonging to different families traditionally used for the treatment of gout and related symptoms by indigenous people of India. The aqueous, methanol-water mixture and methanolic extract of these plants were used for the experiment. Of the 18 extracts assayed, 14 extracts demonstrated xanthine oxidase inhibitory activity at 100 microg/ml, among which 10 extracts showed an inhibition greater than 50% and IC(50) values below 100 microg/ml. The methanolic extracts of Coccinia grandis, Datura metel, Strychnos nux-vomica and Vitex negundo showed more than 50% inhibition, hence, they were screened for their in vivo hypouricaemic activity against potassium oxonate-induced hyperuricaemia in mice. Methanolic extracts of Coccinia grandis and Vitex negundo showed a significant decrease in the serum urate level (3.90+/-0.07 mg/dl, P<0.001) and (6.26+/-0.06 mg/dl, P<0.01), respectively, when compared to hyperuricaemic control (11.42+/-0.14 mg/dl). This effect is almost similar to the serum urate level of allopurinol (3.89+/-0.07 mg/dl).

Animals↗

Ethnopharmacological survey of plants used in the traditional treatment of hypertension and diabetes in south-eastern Morocco (Errachidia province).

This survey was undertaken in the Errachidia province in south-eastern Morocco in order to inventory the main medicinal plants used in folk medicine to treat arterial hypertension and diabetes mellitus. Four hundred individuals who knew about and/or had used the medicinal plants for the indicated diseases, including some herbal healers, were interviewed throughout different regions of the province. The inventory of medicinal plants is summarized in a synoptic table, which contains the scientific, vernacular and common name of the plant, its ecological distribution, the part of the plant and the preparation used and the therapeutic indication. Extensive investigations have brought to light 64 medicinal plants belonging to 33 families; of these, 45 are used for diabetes, 36 for hypertension, and 18 for both diseases. Of these plants, 34% grow in the wild, 44% are cultivated, and 22% are not indigenous to the area and are brought from other parts of Morocco or from outside the country. The survey shows that 78% of the patients regularly use these medicinal plants. In this region, the most frequently used plants to treat diabetes include Ajuga iva, Allium cepa, Artemisia herba-alba, Carum carvi, Lepidium sativum, Nigella sativa, Olea europaea, Peganum harmala, Phoenix dactylifera, Rosmarinus officinalis, and Zygophyllum gaetulum, and those to treat hypertension include Ajuga iva, Allium cepa, Allium sativum, Artemisia herba-alba Asso, Carum carvi, Nigella sativa, Olea europea, Rosmarinus officinalis, Origanum majorana, Peganum harmala, and Phoenix dactylifera. The local people recognize the toxic plants and are very careful in using such plants, which are Citrullus colocynthis, Datura stramonium, Nerium oleander, Nigella sativa, Peganum harmala and Zygophyllum gaetulum. Our survey shows that traditional medicine in the south-eastern Moroccan population has not only survived but has thrived in the transcultural environment and intermixture of many ethnic traditions and beliefs.

Anti-Infective Agents↗

Inhibition of yeast binding to mouse peritoneal macrophages by wheat germ agglutinin: a novel effect of the lectin on phagocytic cells.

The ability of wheat germ agglutinin (WGA) to enhance the binding of bacteria and tumor cells to phagocytic cells, and to induce the killing of tumor cells by macrophages and monocytes, is well established. We observed, however, that WGA inhibits the binding to and phagocytosis of yeast cells by thioglycolate-elicited murine peritoneal macrophages. In order to follow these processes rapidly, the yeasts were labeled with Congo-red and their binding to the macrophages was measured spectrophotometrically after treatment with sodium dodecylsulfate. Phagocytosis was also followed by light microscopy. Binding of the yeasts was inhibited by about 80% after pretreating the macrophages with 150 micrograms/ml of WGA. This effect was reversed by subsequent incubation with N-acetyl-D-glucosamine, chitobiose or chitotriose, but was unaffected by methyl alpha-D-mannoside, N-acetyl-D-mannosamine, D-mannose or D-galactose. Pretreatment of the Congo-red yeasts with WGA did not inhibit their binding by the macrophages. Of a variety of lectins tested, only WGA and Datura stramonium lectin had this effect. Pretreating the macrophages with sialidase prevented the inhibition induced by WGA. Our findings suggest the presence on the macrophages of a class of WGA receptors not previously reported.

Animals↗

Regulation of gelatinase B (MMP-9) in leukocytes by plant lectins.

The stimulatory or inhibitory effects of plant lectins on the production of gelatinase A (MMP-2) and gelatinase B (MMP-9) by mononuclear white blood cells was investigated by substrate zymography. Leukocyte cultures from 24-h old buffy coats were spontaneously activated and produced high levels of gelatinase B. Using such cultures the suppressing activity of the Datura stramonium, Viscum album, Bauhinia purpurea, Triticum aestivum and Maackia amurensis lectins on gelatinase B induction were demonstrated. When fresh leukocyte preparations from single blood donors were used, low levels of gelatinase B were produced. The induction of gelatinase B was confirmed for concanavalin A and phytohaemagglutinin (PHA-L4). In addition, the Urtica dioica, Calystegia sepium, Convolvulus arvensis and Colchicum autumnale lectins were documented as novel and potent inducers of gelatinase B. Since high circulating gelatinase B levels are associated with specific pathologies, including shock syndromes, the acute toxicity of many lectins might be partially mediated or influenced by gelatinase induction.

Cells, Cultured↗

Versatile two-phase solvent system for alkaloid separation by high-speed counter-current chromatography.

In order to find a versatile high speed counter-current chromatography solvent system that can be used as a general prefractionation system for most alkaloids, the crude extracts of five Chinese traditional medicinal herbs, Cortex phellodendri, Semen strychni, green tea, Sophora flavescens ait, and Datura mete L. were resolved. All separations were performed only with a two-phase system composed of CHCl3-CH3OH-water (4:3:2). The water had different acidities controlled by adding NaH2PO4 or HCl to each sample. The fractionated components were identified by thin-layer chromatography, which confirmed this solvent system was versatile and very useful for the separation of alkaloids.

Alkaloids↗

Absorption and storage of phosphorus by larval Manduca sexta.

The role of phosphorus (P) in numerous important biological structures, coupled with the observation that P-content of many insect foods is disproportionately low, suggests that P may be a critical nutrient for growing insects - however, the few studies examining the effects of dietary P on insect performance have generally found only weak relationships. This mismatch may be reconciled by understanding the physiological mechanisms by which insects handle P. Here we describe P processing by larvae of Manduca sexta. When given un-manipulated leaves of a common host plant, Datura wrightii, fifth-instar larvae retained about 85% of P consumed; when given P-enriched leaves larvae retained only 25% of P consumed. Analysis of gut concentrations of P at four sites along the digestive tract, and in leaves and feces, indicates that the rectum is the primary site of P transport between the gut and body and that differences in P retention may be accounted for by differential rates of rectal P transport. Larvae given P-enriched leaves also showed an eightfold increase in the concentration of P in the hemolymph, primarily as alpha-glycerophosphate - but only a 12% increase in the concentration of P in body tissues, suggesting that hemolymph plays a central role in storage and buffering of P.

Journal Article↗

Foraging in nature by larvae of Manduca sexta-influenced by an endogenous oscillation.

Foraging by larvae of the moth Manduca sexta was studied in the field on the native host plant Datura wrightii. Continuous observation of individuals revealed that larvae moved very little, but fed for approximately one third of the time, independent of temperature or photoperiod, although the larger ones fed for the longest times. They were indiscriminate with respect to the part of plant fed upon, and analysis of feeding bouts and interbouts indicated that the pattern of feeding was not strongly influenced by direct nutritional factors. Attacks by parasitoids had a significant impact on feeding behavior. All individuals showed strong regularity in the times when feeding was initiated, and analyses demonstrated that feeding was initiated at times corresponding to fitted oscillation periods of approximately 4 min. Feeds were not initiated on every peak, but at times corresponding to the primary period, or harmonics of it. The period was not influenced by temperature, was different for all individuals, and was asynchronous among individuals. We conclude that an endogenous neural oscillation underlies the rhythm, and strongly influences the overall pattern of foraging. We discuss the oscillation in terms of its possible ecological significance, particularly the need for vigilance.

Journal Article↗

Calystegine distribution in some solanaceous species.

The distribution of eight calystegines (A(3), A(5), B(1), B(2), B(3), B(4), C(1) and N(1)) and their content was investigated by gas chromatography coupled to mass spectrometry (GC-MS) in Datura metel, Atropa belladonna, Hyoscyamus albus, Mandragora autumnalis, Solanum sodomaeum, Withania somnifera, Withania frutescens and Brunfelsia nitida. The most frequently encountered calystegines were A(3), B(1), B(2) and B(3), while distribution of N(1) and C(1) was more limited. In all the investigated samples, calystegines A(5) and B(4) were never detected. This report focuses for the first time on calystegines in Withania and Brunfelsia genera and in Mandragora autumnalis and Solanum sodomaeum species.

Alkaloids↗

Amino acid sequence of ferredoxin from Physalis alkekengi var. francheti.

The complete amino acid sequence of [2Fe-2S] ferredoxin from Physalis alkekengi var. francheti has been determined by automated Edman degradation of the entire Cm-protein and of the peptides obtained by trypsin and endoproteinase Asp-N digestions. This ferredoxin exhibited ten, ten, and nine differences respectively in the amino acid sequence, when compared with the ferredoxins of Datura stramonium, D. metel, and D. arborea, but 21-28 differences for other angiosperms, and 34-37 differences for fern and horsetails. These results are in harmony with the taxonomic position for these plants.

Amino Acid Sequence↗

Lectins in the unicellular Tetrahymena. II. Impact of nutrition and sugar treatment on anti-lectin binding.

Tetrahymena pyriformis GL cells showed partly intracytoplasmic, partly intramembraneous accumulation of lectin-like proteins in conditions of starvation, as demonstrated cytochemically by reaction with antibodies to pea, lens, bean, Datura, and snail lectin. The lectins binding to simple sugars tended to accumulate in the membrane, whereas those capable of interacting with hexosamines in the cytoplasm. While the fluorescence pattern of lectin localization was generally homogeneous in normally nourished cells, it assumed a variegated appearance in starved cells, owing to patching of the membrane, and spot-like accumulation of lectins in the cytoplasm, presumably within membranes of endocytosed vesicles. Snail lectin was an exception, since its distribution remained homogeneous also in conditions of starvation.

Animals↗

Lectins in the unicellular Tetrahymena. I. Lectin detection with FITC-labeled anti-lectins.

FITC-labeled antibodies prepared in rabbits to bean, pea, lens, datura, and snail lectin indicated by immunological reaction presence in the Tetrahymena of corresponding lectin-like components. The latter localized partly on the surface, partly inside the body of the unicellular. The specificity of the anti-lectins was ascertained by immunological and hemagglutination tests, and by examining antibody binding after pretreatment of the Tetrahymena with lectin or sugar.

Animals↗

Retention of endothelium on ovine collagen biomatrix vascular conduits under physiological shear stress.

This study evaluates the adhesion of endothelial cells to 4 mm internal diameter, ovine collagen biomatrix vascular conduits. The biomatrix conduit is formed in a living animal and the wall consists of a complete, naturally produced matrix reinforced with polyester mesh. We propose that the microarchitecture of the matrix lining the lumen may promote endothelial cell attachment without pretreatment with adhesive proteins or extra cellular matrix components. Endothelial cell adhesion to the biomatrix surface was assessed by subjecting conduits seeded with ovine aortic endothelial cells (OAEC) to physiological range shear stresses of 16 and 32 dyn/cm2 in vitro. OAECs were isolated, cultured and seeded (1 x 10(6) cells/ml) by rotation onto the luminal surface of 20 cm lengths of biomatrix vascular conduits (n = 36). The seeded conduits were divided into three groups and cultured either for 24 h (n = 12), 48 h (n = 12) or 72 h (n = 12). Following culture, the conduits from each group were subjected to flow rates of either 240 ml/min (n = 6) or 480 ml/min (n = 6) with heparinized sheep blood for 1 h. Luminal surface cell cover was determined pre- and post-flow from Datura stramonium lectin labeled en face preparations. Histological analysis demonstrated that OAECs attach to the luminal surface of biomatrix conduits and form confluent monolayers within 24-48 h. Flow testing revealed that, for both flow rates and independent of the time in culture, there was no significant decrease in cell cover after flow (p = 0.13). The results support the hypothesis that a vascular conduit, engineered from a naturally formed biomatrix, provides a suitable substrate for the formation of flow resistant endothelium.

Animals↗

Occurrence of poly-N-acetyllactosamine synthesis in Sf-9 cells upon transfection of individual human beta-1,4-galactosyltransferase I, II, III, IV, V and VI cDNAs.

Lectin blot analysis of membrane glycoprotein samples from Sf-9 cells upon transfection of individual human beta-1,4-galactosyltransferase (beta-1,4-GalT) I, II, III, IV, V et VI cDNAs showed that the endogenous N-linked oligosaccharides are galactosylated (Guo et al., Glycobiology (2001), in press). Further analysis revealed that membrane glycoprotein samples from all the gene-transfected cells are also reactive to Lycopersicon esculentum agglutinin (LEA) et Datura stramonium agglutinin (DSA), both of which bind to oligosaccharides with poly-N-acetyllactosamine chains while no lectin reactive protein bands are detected when blots are pretreated with a mixture of diplococcal beta-1,4-galactosidase et jack bean beta-N-acetylhexosaminidase or N-glycanase. Analysis of endo-beta-galactosidase-digestion products revealed the presence of the Gal1-->GlcNAc1-->Gal and/or GlcNAc1-->Gal structures in the gene-transfected cells. When the homogenates of the gene-transfected cells were used as enzyme sources towards oligosaccharides with the GlcNAc beta 1-->(3Gal beta 1-->4GlcNAc)(1-3) structures, human recombinant beta-1,4-GalTs I et II galactosylated these oligosaccharides more effectively than other beta-1,4-GalTs. These results indicate that beta-1,4-GalTs I-VI can synthesize poly-N-acetyllactosamine chains with beta-1,3-N-acetylglucosaminyltransferase.

Animals↗

A rapid method of Sertoli cell isolation by DSA lectin, allowing mitotic analyses.

We have developed a rapid and convenient method of Sertoli cell preparation for studying the growth kinetics of these cells in in vitro culture. Datura Stramonium agglutinin (DSA)-coated dishes were used to rapidly purify single Sertoli cells from immature rat testis. We have monitored by immunohistochemical markers the degree of contamination of our Sertoli cell preparation by other cell types. The cell preparation is essentially free of germ cells and interstitial cells and contains a minimal percentage of myoid cells. Sertoli cells isolated with this method retain functional activities such as the FSH responsiveness in terms of cAMP production. In addition, we have studied the proliferative activity of Sertoli cells isolated by lectin binding from rats of different ages. Sertoli cells exhibited a characteristic pattern of proliferation which was a function of the donor animal age. The proliferative activity of isolated Sertoli cells decreased with age, being much higher in 3 day-old rats than in older animals. A similar pattern was observed when the mitotic activity of Sertoli cells in response to mitogens present in the testicular extracts from 5 day-old rats was evaluated. The method described here reduces or eliminates many of the drawbacks of the conventional procedures used to isolate Sertoli cells, thus providing a useful tool in studies of growth kinetics and regulation of cell proliferation in vitro.

Aging↗

Anticholinergic poisoning from Jimson weed.

Jimson weed (Datura stramonium), is a wild growing herb that contains belladonna alkaloids. Recently there have been reports of intentional ingestion of Jimson weed by adolescents for psychedelic purposes. When seen in emergency department, these patients appear with physical signs of atropine-like poisoning, disturbances of thought and hallucinations. Diagnosis depends on a positive history, if available, and recognition of anticholinergic effects. Differentiation from lysergic acid diethylamide (LSD) ingestion and schizophrenia is important. Physostigmine, an anticholinergic agent, can reverse both central and peripheral manifestations of Jimson weed intoxication.

Belladonna Alkaloids↗

Plant hallucinogens, shamanism and Nazca ceramics.

The ceramics of the ancient Nazca, an extinct people that lived on the south coast of Peru from 100 to 800 AD, are examined. It is suggested that plant hallucinogens and stimulants including Trichocereus pachanoi, Erythroxylon coca, Datura spp., and Anadenanthera peregrina were utilized in religious ritualism connected with shamanism, stressing personal ecstasy as a means of contact with the supernatural on the part of regional religious-political leaders. Shamanic themes linked to world-wide plant hallucinogenic ingestion are identified and summarized, and their representation in Nazca ceramic art delineated.

Ceramics↗

Traditional antihelmintic, antiparasitic and repellent uses of plants in Central Italy.

The uses of 51 plants of Marche, Abruzzo and Latium, distributed in 28 families, are listed here. Memories and news of continued use of the plants in these sectors were collected from farmers and shepherds in person (mostly old people). The plants most frequently used as antiparasitics and repellents are Juglans regia, Lupinus albus, Ruta graveolens, Fraxinus ornus, Datura stramonium, Artemisia absinthium, Allium cepa, while R. graveolens, Cucurbita maxima, A. absinthium, Allium sativum are the most commonly used as antihelmintics.

Animals↗