The modulatory effects of alkaloid extracts of Cannabis sativa, Datura stramonium, Nicotiana tabacum and male Carica papaya on neurotransmitter, neurotrophic and neuroinflammatory systems linked to anxiety and depression.

This study investigated the modulatory effects of alkaloid extracts of Cannabis sativa (CSAE), Datura stramonium (DSAE), Nicotiana tabacum (NTAE) and male Carica papaya (CMAE) on neurotransmitter, neurotrophic and neuro-inflammatory systems linked to anxiety and depression. Male Wistar rats were orally administered the alkaloid extracts in doses of 5, 50, 500, and 2000 mg/kg for 90 days. On day 91, neurobehavioural studies were evaluated, rats were sacrificed, brain hippocampus removed and tissue homogenate prepared. Biochemical, cytokine and neurotransmitter metabolisms were estimated in the hippocampus. Expressions of genes linked to anxiety and depression were evaluated by RT-qPCR. Results showed CSAE, NTAE and CMAE act as anxiolytic and antidepressant agents by depleting TNF-α, IL-1ß and reactive oxygen species concentrations, and monoamine oxidase, angiotensin 1-converting enzyme and acetylcholinesterase activities while elevating IL-10 and dopamine concentrations and glutamate dehydrogenase activity at doses of 5, 50 and 500. Same doses of CSAE, NTAE and CMAE also depleted the gene expressions of GSK3ß, JNK, NF-ĸB, and Nesfatin-1 while increasing expressions of CREB, BDNF, serotonin and Nrf2. However, administration of DSAE and 2000 mg/kg CSAE, NTAE and CMAE had adverse modulatory effects on the neurochemical concentrations and activities as well as the gene expressions of the evaluated neurotransmitter, neurotrophic and inflammatory systems. In conclusion, the study established the sub-chronic instrumentalization potential of CSAE, CMAE, and NTAE for anxiolytic and anti-depressive moods, though their use may be associated with dependence and addiction, which may result in more detrimental effects than any therapeutic potential they may proffer.

Gravimetric quantitative validation of botanic impurities in feed.

BACKGROUND: Harmful botanical impurities may contaminate feed and feed materials and be a potential danger to animal or human health, or to the environment. The aim of this study was to establish rapid and sensitive methods that can be used in routine official controls to determine botanical impurities such as Datura stramonium, Ricinus communis, Crotaliaria spp., and Ambrosia spp. in animal feed and raw materials. Claviceps sclerotia were also detected in cereals, due to the similarities of the targets and the analytical procedure. Regulation (EU) 625/2017, which replaces Reg. 2004/882/EC, states that EU member states should conduct official controls in assessed and accredited laboratories and that the analytical methods must be validated before use by considering parameters such as specificity, precision, recovery, and measurement uncertainly. RESULTS AND CONCLUSION: The results demonstrate that all of the methods tested are suitable for the official quantitative analyses required by EU official legislation. © 2020 Society of Chemical Industry.

Kinetic and thermodynamic studies on biosorption of Cr(VI) on raw and chemically modified Datura stramonium fruit.

Biosorption of Cr(VI) on sulfuric and phosphoric acid-treated Datura stramonium fruit was investigated in batch mode. The various parameters that influence the biosorption process such as Cr(VI) initial concentration, biosorbent dosage, contact time, temperature, and pH value were optimized. Both linear and non-linear regression analysis of isotherm data suggest that Langmuir isotherm model mimics the behavior of Cr(VI) ion biosorption onto Datura stramonium fruit biosorbent. The maximum Cr(VI) ions adsorption capacity of 138.074 mg/g at pH 2 is achieved with phosphoric acid treated Datura stramonium (PDSF). The kinetics of adsorption process is well described by pseudo-second-order model with high R2 and low χ2 value. The estimated activation energy of < 8 kJ/mol obtained for both raw and chemically modified adsorbents suggests that the adsorption occurs mainly via physisorption. Besides, thermodynamic results reveal that biosorption of Cr(VI) on both treated and untreated Datura stramonium was endothermic, spontaneous, and randomness in nature.

Pharmacognostic standardization and DNA fingerprinting of leaves of Datura stramonium, growing naturally in Asir region of Saudi Arabia.

Evaluation of the various pharmacognostic quality parameters and DNA fingerprint of Saudi Arabian medicinal plant namely Datura stramonium growing in Asir region was the objective of the study. The pharmacognostical parameters were done in terms of macroscopic characters, microscopic details, physico-chemical evaluations, phytochemical analysis, fluorescence analysis and DNA fingerprint by using standard techniques and random polymorphic DNA primer. The detailed microscopy of the leaf revealed the presence of pre-medullary phloem, xylem, endodermis, parenchymatous pericycle, lower epidermis and calcium oxalate crystals. There are various amounts of foreign material, ash values, moisture content and extractive values, found after estimations. Preliminary phytochemical screening exhibited the presence of alkaloids, flavonoids, glycosides, tannins and sterols in variable amounts. Random amplified polymorphic DNA (RAPD) showed there are a prominent scorbale DNA bands. These evaluations provided referential information for correct authentication and quality standardization of the important medicinal plant material. These information will also be supportive to differentiate Datura stramonium from the closely related other species.

Elucidating the Distribution of Plant Metabolites from Native Tissues with Laser Desorption Low-Temperature Plasma Mass Spectrometry Imaging.

Secondary metabolites of plants have important biological functions, which often depend on their localization in tissues. Ideally, a fresh untreated material should be directly analyzed to obtain a realistic view of the true sample chemistry. Therefore, there is a large interest for ambient mass-spectrometry-based imaging (MSI) methods. Our aim was to simplify this technology and to find an optimal combination of desorption/ionization principles for a fast ambient MSI of macroscopic plant samples. We coupled a 405 nm continuous wave (CW) ultraviolet (UV) diode laser to a three-dimensionally (3D) printed low-temperature plasma (LTP) probe. By moving the sample with a RepRap-based sampling stage, we could perform imaging of samples up to 16 × 16 cm2. We demonstrate the system performance by mapping mescaline in a San Pedro cactus ( Echinopsis pachanoi) cross section, tropane alkaloids in jimsonweed ( Datura stramonium) fruits and seeds, and nicotine in tobacco ( Nicotiana tabacum) seedlings. In all cases, the anatomical regions of enriched compound concentrations were correctly depicted. The modular design of the laser desorption (LD)-LTP MSI platform, which is mainly assembled from commercial and 3D-printed components, facilitates its adoption by other research groups. The use of the CW-UV laser for desorption enables fast imaging measurements. A complete tobacco seedling with an image size of 9.2 × 15.0 mm2 was analyzed at a pixel size of 100 × 100 µm2 (14 043 mass scans), in less than 2 h. Natural products can be measured directly from native tissues, which inspires a broad use of LD-LTP MSI in plant chemistry studies.

Effect of different solvent extracted samples from the leaves and fruits of Datura stramonium on the growth of bacteria and fungi.

Different solvent extracted samples from the leaves and fruits tissues of D. stramonium were tested against five pathogenic microorganisms by disc diffusion susceptibility method using 1, 2 and 3mg disc-1 concentrations. Methanol and chloroform extracted fractions from both leaves and fruits measured good growth inhibition of all the tested microorganisms at all concentrations. Bacillus subtilis was very resistant to n-butanol and aqueous extracted fractions of fruits tissues at all the tested three concentrations. The growths of Pseudomonas aeruginosa and Klebsiella pnuemonia were efficiently reduced by all the solvent extracted fractions from the fruits while aqueous fraction was unable to restrain the growth of Bacillus subtilis. The growth of Candida albicans was effectively reduced by aqueous extracted fraction from the leaves tissue at the highest concentration. Maximum growth reduction of (77%) was shown by chloroform extracted fractions from the leaves against Klebsiella pneumonia at 3mg disc-1 concentration. Minimum zone of inhibition (35.4%) was measured by n-butanol extracted fractions from the leaves against Pseudomonas aeruginosa at the lowest concentrations of 1mg disc-1. In case of leaves the most vulnerable bacteria was Bacillus subtilis while in case of fruits Klebsiella pneumonia was the most susceptible while Bacillus subtilis was the most resistant one.

Cytotoxicity sesquiterpenoids from the leaves of Datura stramonium L.

Four new sesquiterpenoids, dstramonins A-D (1-4), and one new natural product (5), together with three known compounds (6-8), were isolated from the leaves of Datura stramonium L. The structures of new compounds were elucidated by extensive spectroscopic analysis and comparison with the literature. The cytotoxicity of isolates against LN229 cells was assessed and compounds 2-4, and 7 displayed cytotoxic activity with IC50 values ranging from 8.03 to 13.83 µM.

Screening of immunomodulatory activity of total and protein extracts of some Moroccan medicinal plants.

Herbal and traditional medicines are being widely used in practice in many countries for their benefits of treating different ailments. A large number of plants in Morocco were used in folk medicine to treat immune-related disorders. The objective of this study is to evaluate the immunomodulatory activity of protein extracts (PEs) of 14 Moroccan medicinal plants. This activity was tested on the proliferation of immune cells. The prepared total and PEs of the plant samples were tested using MTT (3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide) assay on the splenocytes with or without stimulation by concanavalin-A (Con-A), a mitogenic agent used as positive control. The results of this study indicated different activity spectra. Three groups of activities were observed. The first group represented by Citrullus colocynthis, Urtica dioica, Elettaria cardamomum, Capparis spinosa and Piper cubeba showed a significant immunosuppressive activity. The second group that showed a significant immunostimulatory activity was represented by Aristolochia longa, Datura stramonium, Marrubium vulgare, Sinapis nigra, Delphynium staphysagria, Lepidium sativum, Ammi visnaga and Tetraclinis articulata. The rest of the plant extracts did not alter the proliferation induced by Con-A. This result was more important for the PE than for the total extract. In conclusion, this study revealed an interesting immunomodulating action of certain PEs, which could explain their traditional use. The results of this study may also have implications in therapeutic treatment of infections, such as prophylactic and adjuvant with cancer chemotherapy.

[Effects of methyl jasmonate on accumulation and release of main tropane alkaloids in liquid cultures of Datura stramonium hairy root].

OBJECTIVE: To study the effects of methyl jasmonate (MJ) on the accumulation and release of main secondary metabolites i. e. scopolamine and hyoscyamine in liquid cultures of Datura stramonium hairy roots. METHOD: After 18 days liquid culture of D. stramonium hairy roots induced by agrobacterium rhizogenes C58C1, the chemical elicitor methyl jasmonate was added into 1/2 MS liquid cultures and scopolamine and hyoscyamine on the day 0, 3, 6, 9 and 12, after dealing with MJ, was determined by HPLC. RESULT: After dealing with MJ on the day 3, 6, 9 and 12,the concentration of scopolamine reached to 0.419, 0.439, 0.431, 0.374 mg x g(-1), respectively, the increase of scopolamine were 1.36, 1.42, 1.17 and 1.12 fold higher than that of the control, respectively. And hyoscyamine reached 1.493, 0.817, 0.723 and 0.698 mg x g(-1), respectively, the increase of hyoscyamine were 2.28, 1.11, 0.63 and 0.70 fold higher than that of the control, respectively. CONCLUSION: MJ could stimulate the accumulation of scopolamine and hyoscyamine (3,6 d) in D. stramonium hairy root and have released them into the culture medium.

Three new α-glucosidase inhibitor benzo-isochromenes from Datura stramonium.

Three new α-glucosidase inhibitory benzo-isochromenes were isolated from the chloroform fraction of Datura stramonium. Their structures were established with the help of modern spectroscopic techniques and were assigned the names as 1,6,8-triimethoxy-2-methyl-3,4-dihydrobenzoisochromene-5,10-diol, 3,6-dimethoxy-5-hydroxy-4-methylbenzoisochromene-9,11-dione and demethylflavasperon for compounds 1-3 respectively. The α-glucosidase inhibiting activity of compound 2 showed strong inhibition with an IC50 value of 27.5 µM, while compound 1 exhibited moderate activity with IC50 value of 60.2 µM compared to positive control (acarbose).

Short-term in-vitro culture of goat enriched spermatogonial stem cells using different serum concentrations.

PURPOSE: To investigate the effect of serum supplementing on short-term culture, fate determination and gene expression of goat spermatogonial stem cells (SSCs). METHODS: Crude testicular cells were plated over Datura-Stramonium Agglutinin (DSA) for 1 h, and non-adhering cells were cultured in the presence of different serum concentrations (1, 5, 10, and 15%) for 7 days in a highly enriched medium initially developed in mice. Colonies developed in each group were used for the assessment of morphology, immunocytochemistry, and gene expression. RESULTS: Brief incubation of testicular cells with DSA resulted in a significant increase in the number of cells that expressed the germ cell marker (VASA). The expression of THY1, a specific marker of undifferentiated spermatogonia, was significantly higher in colonies developed in the presence of 1% rather than 5, 10 and 15% serum. CONCLUSION: Goat SSCs could proliferate and maintain in SSC culture media for 1 week at serum concentrations as low as 1%, while higher concentrations had detrimental effects on SSC culture/expansion.

[Chemical constituents of Datura stramonium seeds].

OBJECTIVE: To study chemical constituents in the seeds of Datura Stramonium (Solanaceae family). METHOD: Compounds were isolated and purified by silica gel, MCI and Sephadex LH-20 column chromatography, and their structures were determined based on physicochemical constants and spectroscopic analysis including NMR and MS. RESULT: Twelve compounds were isolated and identified from Datura stramonium, they were N-trans-feruloyl tryptamine (1), hyoscyamilactol (2), scopoletin (3), umckalin (4), daturaolone (5), daturadiol (6), N-trans-ferulicacyl- tyramine (7), cleomiscosin A (8), fraxetin (9), scopolamine (10), 1-Acetyl-7-hydrox-beta-carbol-ine (11), 7-hydroxy-beta-carbolinel-propionic acid (12). CONCLUSION: Compound 2, 7, 9 and 12 were obtained from Datura genus for the first time, whereas compound 1, 4, 8 and 11 were obtained from the Solanaceae family for the first time.

Mantuoluosides A-H, new steroids isolated from the leaves of Datura stramonium L.

Eight new steroids, designated mantuoluosides A-H (1-8), were obtained from the enrichment of steroids of the Datura stramonium L. using HPD-BJQH macroporous resin. Their respective structures were elucidated based on spectroscopic methods and comparison data with literature. The anti-inflammatory activities of these compounds were evaluated on (LPS)-induced RAW264.7 cells with inhibition ratio. It was found that most isolates showed potential anti-inflammatory activity with varying degrees of inhibition rate in a concentration-dependent manner at non-cytotoxic concentrations.

Evaluation of analgesic and anti-inflammatory activities and molecular docking analysis of steroidal lactones from Datura stramonium L.

BACKGROUND: Datura stramonium L. is widely used across the world for its therapeutic potential to treat inflammatory disorders. The current work was designed to isolate and identify steroidal lactones from D. stramonium leaves and evaluate their anti-inflammatory and analgesic properties. METHODS: Several compounds were isolated from D. stramonium leaves and characterized by nuclear magnetic resonance and high-resonance electron spray ionization mass spectrometry techniques. Further, anti-inflammatory properties of these compounds were evaluated by in vitro assays, such as release of NO and pro-inflammatory cytokines by lipopolysaccharide (LPS)-activated J774A.1 macrophages. Using in vivo models, anti-inflammatory and analgesic effects were examined by mouse tail-flick, carrageenan-induced inflammation in rat paw model, vascular permeability in rats, and acetic acid-induced writhing in mice. The docking studies were performed for assessing the binding efficiency of the test compounds with cyclooxygenase-1 (COX-1) and COX-2, lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), inducible nitric oxide synthases (iNOS) and nuclear factor-κB (NF-κB). RESULTS: Three lactones were isolated and confirmed as daturalactone (D1), 12-deoxywithastramonolide (D23), and daturilin (D27). Further, the isolated compounds showed nitric oxide inhibition and pro-inflammatory cytokines released by LPS-activated J774A.1 macrophages. The in vivo results suggest that D1, D23 and D27 (20 mg/kg) were able to reduce the pain and inflammation in various animal models. The docking analysis showed that these three compounds actively bind with COX-1, COX-2, LOX-1, NF-κB, and iNOS, validating the anti-inflammatory effects of the lactones. CONCLUSION: These findings demonstrate substantial anti-inflammatory and analgesic properties of D. stramonium-derived lactones and their potential as anti-inflammatory agents to treat chronic inflammatory ailments.

Enantiomeric differentiation of atropine/hyoscyamine by (13) C NMR spectroscopy and its application to Datura stramonium extract.

INTRODUCTION: The two enantiomers of hyoscyamine, an alkaloid found in many plant species, have distinct pharmacological and biological properties. Methods for the discrimination of both enantiomers are almost exclusively based on chiral HPLC/UV. Determination of the enantiomeric ratio (e.r.) of hyoscyamine is a challenging problem since this compound tends to racaemise, forming atropine during acid-base extraction. OBJECTIVE: To develop a protocol for the calculation of enantiomeric ratio of hyoscyamine in a plant extract using a (13) C NMR method. METHODOLOGY: Samples were prepared by extraction of dried Datura stramonium seeds. Observation of C12 and C15 NMR signals of hyoscyamine in the presence of one equivalent of TFA and sub-stoichiometric amount of Yb(hfc)(3) allowed the calculation of the e.r. of S-(-) and R-(+)-hyoscyamine.The method was optimised with various mixtures of (+) and (-)-hyoscyamine ranging from 50:50 (racaemic mixture, i.e. atropine) to 98.5:1.5. The e.r. measured by NMR on the signals of aromatic C12 and C15 were in agreement with the gravimetrically prepared samples. The method was then applied to an extract of Datura stramonium and S-(-)-hyoscyamine was the unique enantiomer. CONCLUSION: The study showed that the e.r. determination of atropine/hyoscyamine was achieved with a routine NMR spectrometer, using CLSR/TFA on pure compounds as well as on the crude extract of Datura stramonium.

Datura stramonium intake: a report on three cases.

This article describes three cases of Datura stramonium intake on two nonconsecutive days. In the first case, the patient took a small amount of D. stramonium seeds without showing any symptoms of intoxication. The other two patients had taken a considerable amount of seeds and reported a sudden surge in strength and energy, with some aggressive compulsion towards their peers. They showed delirium as well as confusion and disorientation. The absence of any specific legislation makes D. stramonium a tempting alternative to other psychoactive substances. Thus, it is extremely important to be able to recognize its symptoms so as to be able to diagnose any signs of intoxication properly.

Optimization of the culture medium composition to improve the production of hyoscyamine in elicited Datura stramonium L. hairy roots using the Response Surface Methodology (RSM).

Traditionally, optimization in biological analyses has been carried out by monitoring the influence of one factor at a time; this technique is called one-variable-at-a-time. The disadvantage of this technique is that it does not include any interactive effects among the variables studied and requires a large number of experiments. Therefore, in recent years, the Response Surface Methodology (RSM) has become the most popular optimization method. It is an effective mathematical and statistical technique which has been widely used in optimization studies with minimal experimental trials where interactive factors may be involved. This present study follows on from our previous work, where RSM was used to optimize the B5 medium composition in [NO(3-)], [Ca(2+)] and sucrose to attain the best production of hyoscyamine (HS) from the hairy roots (HRs) of Datura stramonium elicited by Jasmonic Acid (JA). The present paper focuses on the use of the RSM in biological studies, such as plant material, to establish a predictive model with the planning of experiments, analysis of the model, diagnostics and adjustment for the accuracy of the model. With the RSM, only 20 experiments were necessary to determine optimal concentrations. The model could be employed to carry out interpolations and predict the response to elicitation. Applying this model, the optimization of the HS level was 212.7% for the elicited HRs of Datura stramonium, cultured in B5-OP medium (optimized), in comparison with elicited HRs cultured in B5 medium (control). The optimal concentrations, under experimental conditions, were determined to be: 79.1 mM [NO(3-)], 11.4 mM [Ca(2+)] and 42.9 mg/L of sucrose.

Genomic and chemical evidence for local adaptation in resistance to different herbivores in Datura stramonium.

Because most species are collections of genetically variable populations distributed to habitats differing in their abiotic/biotic environmental factors and community composition, the pattern and strength of natural selection imposed by species on each other's traits are also expected to be highly spatially variable. Here, we used genomic and quantitative genetic approaches to understand how spatially variable selection operates on the genetic basis of plant defenses to herbivores. To this end, an F2 progeny was generated by crossing Datura stramonium (Solanaceae) parents from two populations differing in their level of chemical defense. This F2 progeny was reciprocally transplanted into the parental plants' habitats and by measuring the identity by descent (IBD) relationship of each F2 plant to each parent, we were able to elucidate how spatially variable selection imposed by herbivores operated on the genetic background (IBD) of resistance to herbivory, promoting local adaptation. The results highlight that plants possessing the highest total alkaloid concentrations (sum of all alkaloid classes) were not the most well-defended or fit. Instead, specific alkaloids and their linked loci/alleles were favored by selection imposed by different herbivores. This has led to population differentiation in plant defenses and thus, to local adaptation driven by plant-herbivore interactions.

Evaluation of Two New Methods for DNA Extraction of "Legal High" Plant Species.

A quick, simple, and high-yield nucleic acid isolation process is crucial for high-quality DNA analysis. The ability of the MicroGEM PDQeX phytoGEM system and Omega Bio-tek E.Z.N.A.® Plant DS Mini kit to extract PCR-ready DNA was evaluated by extracting the forensically relevant "legal high" plant species: Ipomoea purpurea, Artemisia absinthium, Mitragyna speciosa, Datura stramonium, and Papaver somniferum. The plant material was pulverized, processed using the manufacturer's plant protocol for the PDQeX Nucleic Acid Extraction or the manufacturer's protocol for the Omega extraction, quantified using the Invitrogen Qubit 2.0 Fluorometer, and analyzed for amplifiability by PCR using a Qiagen Rotor-Gene Q instrument and published assays. The DNA amplicons for the legal high species produced high-resolution melt curves concordant with the melts observed when DNA was isolated using the Qiagen DNeasy Plant Mini Kit in previous studies.

Combined lectin- and immuno-histochemistry (CLIH) for applications in cell biology and cancer diagnosis: Analysis of human urothelial carcinomas.

Lectin histochemistry (LHC) and immunohistochemistry (IHC), which demonstrate the composition and localisation of sugar residues and proteins in cell membranes, respectively, are generally used separately. Using these two methods, we previously demonstrated that malignant transformation of urothelial cells results in the alterations of protein glycosylation and reduced expression of urothelium-specific integral membrane proteins uroplakins (UPs). However, the correlation between these changes was not studied yet. To evaluate this correlation, we developed innovative method, which we named combined lectin- and immuno- histochemistry (CLIH). We used human biopsies of 6 normal urothelia and 9 papillary urothelial carcinomas, i.e. 3 papillary urothelial neoplasms of low malignant potential (PUNLMP), 3 non-invasive papillary urothelial carcinomas of low grade (pTa, l.g.), and 3 invasive papillary urothelial carcinomas of high grade (pT1, h.g.). We tested five different protocols (numbered 1-5) of CLIH on paraffin and cryo-semithin sections and compared them with LHC and IHC performed separately. Additionally, we carried out western and lectin blotting with antibodies against UPs and lectins Amaranthus caudatus agglutinin (ACA), Datura stramonium agglutinin (DSA), and jacalin, respectively. We showed that incubation with primary antibodies first, followed by the mixture of secondary antibodies and lectins is the most efficient CLIH method (protocol number 5). Additionally, 300 nm thick cryo-semithin sections enabled better resolution of co-localisation between sugar residues and proteins than 5 µm thick paraffin sections. In the normal urothelium, CLIH showed co-localisation of lectins ACA and jacalin with UPs in the apical plasma membrane (PM) of superficial umbrella cells. In papillary urothelial carcinomas, all three lectins (ACA, DSA and jacalin) labelled regions of apical PM, where they occasionally co-localised with UPs. Western and lectin blotting confirmed the differences between normal urothelium and papillary urothelial carcinomas. Our results show that CLIH, when used with various sets of lectins and antigens, is a useful, quick, and reliable method that could be applied for basic cell biology research as well as detailed subtyping of human urothelial carcinomas.