Lectin histochemical studies on the olfactory gland and two types of gland in vomeronasal organ of the brown bear.[Pubmed: 30001800]

Olfaction is mediated by the vomeronasal and main olfactory systems, and the peripheral vomeronasal organ (VNO) processes species-specific chemicals that are associated with various behaviors in mammals. Sensory epithelial surfaces of the olfactory mucosa and VNO are covered by mucosal fluid that contains secretory products derived from associated glands, and glycoconjugates in the mucosal fluid are involved in odorant reception. The VNO of brown bears contains two types of glands; submucosal vomeronasal glands (VNG) and multicellular intraepithelial glands (MIG). The present study determined the labelling profiles of 21 lectins in the olfactory glands (OG), VNG and MIG of young male brown bears. The OG reacted with 12 lectins, and the VNG and MIG were positive for seven and eight lectins, respectively. Six lectins bound only to the OG, while four reacted with both or either of the VNG and MIG, but not the OG. The differences of lectin labelling pattern between the OG and glands in the VNO suggest that glycans in covering mucosal fluids differ between the olfactory mucosa and VNO. In addition, Bandeiraea simplicifolia lectin-I, Sophora japonica agglutinin and Jacalin reacted with the MIG but not the VNG, whereas Datura stramonium lectin and concanavalin A bound to the VNG, but not the MIG. These findings indicate that the properties of secretory substances differ between the two types of glands in the bear VNO, and that the various secretions from these two types of glands may function in the lumen of VNO together.

A Rapid, High-Throughput Validated Method for the Quantification of Atropine in Datura stramonium Seeds Using Direct Analysis in Real Time-High Resolution Mass Spectrometry (DART-HRMS).[Pubmed: 29974430]

The utility of direct analysis in real time-high resolution mass spectrometry (DART-HRMS) for quantification of a variety of compounds has been explored, but the number of reports of validated methods using this technique is limited. Furthermore, despite the increasing use in crime labs of DART-HRMS for the detection and identification of drugs of abuse, very few published reports have appeared describing how the method can be exploited for the analysis of small molecules of interest within complex matrices such as plant tissues. Herein we describe the steps to be taken to establish a validated quantification method for psychoactive compounds within complex plant matrices through its application to the detection and quantification of atropine in Datura stramonium seeds. Six calibration standard series are analyzed eight times over a period of several days to create a calibration curve. The resulting calibration curve is tested using six quality control samples and finally utilized to determine the concentration of atropine in a D. stramonium seed extract. The linear range for quantification of atropine in this study was found to be comparable to that reported previously using GC, LC, HPLC, and UHPLC-MS methods. Furthermore, the method can be applied to the quantification of other biomarkers in plant materials, despite the complexity of the plant matrix. The speed of the analysis (<10 min for duplicate analysis of 20 samples) and the ability to integrate peaks using accurate masses for specificity are advantages of the DART-HRMS quantification approach.

Simultaneous analysis of tropane alkaloids in teas and herbal teas by liquid chromatography coupled to high-resolution mass spectrometry (Orbitrap).[Pubmed: 29359500]

A new method has been developed for the simultaneous determination of 13 tropane alkaloids in tea and herbal teas using high-performance liquid chromatography coupled to an Exactive-Orbitrap analyzer. A mixture of methanol, water, and formic acid was used for the extraction of the target compounds followed by a solid-phase extraction step. The validated method provided recoveries from 75 to 128% with intra- and interday precision lower than or equal to 24% (except for apoatropine). Limits of quantification ranged from 5 to 20 μg/kg. Eleven tea and herbal tea samples and two contaminated samples with Datura stramonium seeds were analyzed. Tropane alkaloids were detected in six samples with concentrations from 5 (apoatropine) to 4340 μg/kg (sum of physoperuvine, pseudotropine, and tropine), whereas concentrations from 5 (apoatropine) to 1725 μg/kg (sum of physoperuvine, pseudotropine, and tropine) were found in the contaminated samples.

Toxic effect of common poisonous plants of district Bannu, Khyber Pakhtunkhwa, Pakistan.[Pubmed: 29348085]

The present paper was a part of Ph.D research work, conducted during the year 2014, in which 87 poisonous plants belonging to 54 genera, were collected, documented and preserved in the herbarium of Bannu, Department of Botany UST, Bannu Khyber Pakhtunkwa Pakistan. The plants were identified botanically, arranged alphabetically along with their Latin name, family name, common name, poisonous parts, toxicity, affects, toxin and their effects. Aim of the study was to induce awareness in the local people of district Bannu about the poisonous effects of the commonly used plants. Data about poisonous effect were collected from the local experienced and mostly old age people through questionnaire. Some information were collected from a number of veterinary texts and literature. The most important plants genera studied in the area were Brassica 6 species (11.11%), Lathyrus 5 spp (9.26%), Astragalus, Euphorbia and Prunus were with 4 spp (7.40%). Datura, Jatropha, Ranunculus, Solanum and Sorghum were with 3 spp (5.56%) while Allium, Amaranthus, Chenopodium, Melilotus and Taxus were with 2 spp (3.70%). These 15 genera contribute 48 species (55.17 %) while the remaining 39 genera have single species each and contribute 44.83% to the total poisonous flora of the research area. Other important poisonous plants were Anagallis arvensis L., Cannabis sativa, Datura stramonium L., D. metel L., Euphorbia species, Heliotropium europaeum, Ipomoea tricolor, Jatropha curcas, Lolium temulentum L., Malus domestica, Mangifera indica L., Medicago sativa L., Melilotus alba Desr., M. officinalis (L.) Lam., Mirabilis jalapa L., Narcissus tazetta, Nicotiana tabacum L., Sorghum halepense (L) Pers., and Xanthium strumarium. It was concluded that the local population had poor knowledge about the poisonous effect of the plants and the present research work was anticipated for use by health care professionals, veterinarians, farmers, homeowners, as well as botanically curious individuals.

Lectins identify distinct populations of coelomocytes in Strongylocentrotus purpuratus.[Pubmed: 29125863]

Coelomocytes represent the immune cells of echinoderms, but detailed knowledge about their roles during immune responses is very limited. One major challenge for studying coelomocyte biology is the lack of reagents to identify and purify distinct populations defined by objective molecular markers rather than by morphology-based classifications that are subjective at times. Glycosylation patterns are known to differ significantly between cell types in vertebrates, and furthermore they can vary depending on the developmental stage and activation states within a given lineage. Thus fluorescently labeled lectins that recognize distinct glycan structures on cell surface proteins are routinely used to identify discrete cell populations in the vertebrate immune system. Here we now employed a panel of fifteen fluorescently-labeled lectins to determine differences in the glycosylation features on the surface of Strongylocentrotus purpuratus coelomocytes by fluorescence microscopy and flow cytometry. Eight of the lectins (succinylated wheat germ agglutinin, Len culinaris lectin, Pisum sativum agglutinin, Saphora japonica agglutinin, Solanum tuberosum lectin, Lycopersicon esculentum lectin, Datura stramonium lectin, Vicia villosa lectin) showed distinct binding patterns to fixed and live cells of three major coelomocyte classes: phagocytic cells, red spherule cells, and vibratile cells. Importantly, almost all lectins bound only to a subgroup of cells within each cell type. Lastly, we established fluorescently-labeled lectin-based fluorescence activated cell sorting as a strategy to purify distinct S. purpuratus coelomocyte (sub-)populations based on molecular markers. We anticipate that this will become a routine approach in future studies focused on dissecting the roles of different coelomocytes in echinoderm immunity.