Radioiodination and Purification of [131I]β-CIT and [131I]FP-CIT with an Automated Radiosynthesizer

Dopaminergic transporter (DAT) imaging with single photon emission computed tomography (SPECT) is used to diagnose Parkinson’s disease and to differentiate it from other neurodegenerative disorders without presynaptic dopaminergic dysfunction. The radioiodinated tropane alkaloids [123I]FP-CIT and [123I]β-CIT enable the evaluation of the integrity of DATs. Commonly, the labeling of these compounds is performed by electrophilic substitution of the alkylstannylated precursors with radioactive iodine and following purification by HPLC or solid phase extraction (SPE). This work presents the first radioiodination of β-CIT and FP-CIT with no carrier added [131I]NaI on a Scintomics GRP synthesis module. Free iodine-131 and impurities were removed by SPE over a C-18 Sep-Pak cartridge. We achieved a radiochemical yield of >75% and a radiochemical purity of >98% with both compounds. Our development of an automated synthesis on a commercially available synthesizer ensures robust and efficient labeling of [131I]FP-CIT and [131I]β-CIT starting with low concentrated radioiodine.

Next-generation biomonitoring of the early-life chemical exposome in neonatal and infant development

Exposure to man-made and natural chemicals is a major, yet not sufficiently considered, environmental risk factor in the etiology of chronic diseases. Current human biomonitoring approaches typically measure a limited number of exposures rather than investigating complex mixtures. The latter would be fundamental and necessary for a holistic assessment of chemical exposure in exposome-wide association studies. In this work, an highly-sensitive liquid chromatography-tandem mass spectrometry approach was developed and thoroughly-validated. The assay enables the simultaneous and targeted assessment of more than 80 highly-diverse xenobiotics in the investigated body fluids of urine, serum/plasma, and breast milk; the detection limit for most toxicants are in the pg-ng/mL range. In the plasma of extremely-premature infants (gestational age <28 weeks, birth weight <1 kg) a total of 27 different xenobiotics are identified; including severe contamination with synthetic plasticizers, perfluorinated alkylated substances and parabens. In an independent sample set of breast milk that was longitudinally collected over the first 211 days post-partum, a total of 29 analytes is detected, including the first-ever identification of pyrrolizidine- and tropane alkaloids in this matrix. Based on the generated data, a preliminary estimation of daily toxicant intake via breast milk is conducted. In conclusion, our proof-of-principle experiments show significant early-life co-exposure to multiple toxicants, and demonstrate the method’s applicability in future large-scale exposomics-type cohort studies in vulnerable populations.

Pipes and Potions: Testing the Efficacy of European Folk Preparation Methods for Anticholinergic Solanaceae Plants

The present article sought to evaluate the efficiency of various folk preparation methods commonly used in Europe for employing anticholinergic Solanaceae plants. The study aimed to uncover which folk methods were effective for the extraction of the anticholinergic tropane alkaloids of these plants, atropine and scopolamine. The folk extractions that were tested sought to simulate the preparation of teas, cold-water infusions, unguents, tinctures, fortified wines, and smoking. All preparation types and a control were then put through an extraction process to see what amount of the alkaloids had been maintained. These extractions were then analysed using high-performance liquid chromatography (HPLC). Cold- and hot-water preparations, tinctures, and fortified wines all proved to be effective means of extracting atropine and scopolamine from plant material under conditions seen in folk usage. Smoking and the oil-based unguent, however, yielded no alkaloids, suggesting a lack of efficiency for these preparations, a problem with our methodology, or possible chemical changes and losses associated with the preparation procedure.

The history, current state and perspectives of aerosol therapy

Nebulization is a very effective method of drug administration. This technique has been popular since ancient times when inhalation of plants rich in tropane alkaloids with spasmolytic and analgesic effects was widely used. Undoubtedly, the invention of anasthesia in the 19th century had an influence on the development of this technique. It resulted in the search for devices that facilitated anasthesia such as pulveriser or hydronium. From the second half of the 21st century, when the first DPI and MDI inhalers were launched, the constant development of aerosol therapy has been noticed. This is due to the fact that nebulization, compared with other means of medicinal substance application (such as oral and intravenous routes of administration), is safer and it exhibits a positive dose/efficacy ratio connected to the reduction of the dose. It enables drugs administration through the lung and possesses very fast onset action. Therefore, various drugs prescribed in respiratory diseases (such as corticosteroids, β-agonists, anticholinergics) are present on the market in a form of an aerosol.

The Effect of Simulated Microgravity On The Antioxidant Capacity And Alkaloid Formation In The Hyoscyamus Niger

Microgravity is one of the most important abiotic stresses in space. In the case of plant exposure to short term microgravity, plants establish strategies to response to these stresses and promote growth and survival. We hypothesized that the simulated microgravity can promote the antioxidant capacity and the formation of secondary metabolites such as tropane alkaloids in the Hyoscyamus niger. Callus induction was conducted by putting hypocotyl segments of H. niger seedlings in solid MS medium supplemented with 1 mg L−1 2,4-D and 1 mg L−1 BAP growth regulators. Then, the sub-cultured calli were placed on a clinostat for 3, 7 and 10 days. We performed Atropine and Scopolamine determination through HPLC. PAL (Phenyle alanine amonalyase) and antioxidant activity were also determined. Gene expression analysis of jasmonic acid (JA), Hyoscyamine 6-beta Hydroxylase (H6H), Putrescine N-methyltransferase (PMT), mitogen-activated protein kinase (MAPK) and ethylene responsive element binding (EREB) was performed using quantitative real time PCR. Findings showed that microgravity had a positive effect on the antioxidant capacity, Atropine and Scopolamine production in the H. niger calli. However, microgravity had a negative effect on the PAL activity. Furthermore, gene expression analysis indicated that microgravity significantly induced gene expression of the H6H, PMT and JA. It was also revealed that callus growth, carbohydrate and protein content increased in response to microgravity treatment. We conclude that microgravity can be considered as a potent factor to induce plant antioxidant activity and tropane alkaloids formation to be applicable in the pharmaceutical and medicinal industries.

Datura stramonium Leaf Extract Toxic Effects on Testis in Swiss Albino Mice Mus muscullu

The notorious weed, jimson weed (Datura stramonium L.) is a hallucinating plant with both poisonous and medicinal properties. The neurotoxicity of the plant is attributed to the presence of tropane alkaloids which contain a methylated nitrogen atom (N-CH3) that block neurotransmitters in the brain. Ethno-medicinally, the frequent recreational abuse of D. stramonium has resulted in toxic syndromes. This investigation has been designed to examine the toxicity and describe the possible changes in the structural function of vital organs, following the oral intubation of non-lethal doses of Datura stramonium leaves crude aqueous extract. Through preliminary trials, crude aqueous extract. Of 200mg leaves per kilogram body weight was established as a tolerable non-lethal dose. Three doses 0.36, 0.7, and 4 mg/kg were orally weekly, administered to the male mice in a 0.1 ml volume. Acute toxicity studies were accomplished through oral intubation of three dosages in each case. Observation and mortality were reported for 24 .48, 72 hours Prolonged toxicity was performed through the administration of weekly, single doses oral for 40 days. The observation was made on the mice's body weight, , and histological abnormality of a testis organ.

CRISPR/Cas9-mediated disruption of the PYRROLIDINE KETIDE SYNTHASE gene reduces the accumulation of tropane alkaloids in Atropa belladonna hairy roots

Tropane alkaloids, including clinically important hyoscyamine and scopolamine, are produced in the roots of medicinal plant species, such as Atropa belladonna, from the Solanaceae family. Recent molecular and genomic approaches have advanced our understanding of the metabolic enzymes involved in tropane alkaloid biosynthesis. A non-canonical type III polyketide synthase (PKS), pyrrolidine ketide synthase (PYKS), catalyzes a two-step decarboxylative reaction, which involves imine-ketide condensation indispensable to tropane skeleton construction. In this study, we generated pyks mutant A. belladonna hairy roots via CRISPR/Cas9-mediated genome editing and analyzed the metabolic consequences of the loss of PYKS activity on tropane alkaloids, providing insights into a crucial role of the scaffold-forming reaction in the biosynthetic pathway.

Multivariate statistical analysis of tropane alkaloids in Anisodus tanguticus (Maxim.) Pascher from different regions to trace geographical origins

Anisodus tanguticus (Maxim.) Pascher is an important Tibetan folk medicine and the source of tropane alkaloids (TAs) grown in Qinghai-Tibet Plateau. There are marked differences in quality of A. tanguticus from geographic areas. The aim of present research was to establish a method for the quantitative analysis of TAs coupled with chemometrics analysis to trace geographical origins. Qualitative analysis of TAs in A. tanguticus was carried out using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry and quantitative analysis of TAs in different plant organs from different geographical origin was achieved. Contents of TAs were subjected to the principal component analysis, and orthogonal partial least-squares discriminant analysis. The contents of the three marker compounds (anisodamine, anisodine and atropine) in the roots and acrial parts of A. tanguticus were positive correlated and varied significantly from different geographical origins. Principal component analysis, and orthogonal partial least-squares discriminant analysis results showed excellent discrimination between different geographical origin of A. tanguticus. This study could provide comprehensive evaluation and further utilization of A. tanguticus resources.

Development of CE-C4D Method for Determination Tropane Alkaloids

A fast method for the determination of tropane alkaloids, using a portable CE instrument with a capacitively coupled contactless conductivity detector (CE-C4D) was developed and validated for determination of atropine and scopolamine in seeds from Solanaceae family plants. Separation was obtained within 5 min, using an optimized background electrolyte consisting of 0.5 M acetic acid with 0.25% (w/v) β-CD. The limit of detection and quantification was 0.5 µg/mL and 1.5 µg/mL, respectively, for both atropine and scopolamine. The developed method was validated with the following parameters—precision (CV): 1.07–2.08%, accuracy of the assay (recovery, RE): 101.0–102.7% and matrix effect (ME): 92.99–94.23%. Moreover, the optimized CE-C4D method was applied to the analysis of plant extracts and pharmaceuticals, proving its applicability and accuracy.