Bioprospecting of Artemisia genus: from artemisinin to other potentially bioactive compounds.

Species from genus Artemisia are widely distributed throughout temperate regions of the northern hemisphere and many cultures have a long-standing traditional use of these plants as herbal remedies, liquors, cosmetics, spices, etc. Nowadays, the discovery of new plant-derived products to be used as food supplements or drugs has been pushed by the exploitation of bioprospection approaches. Often driven by the knowledge derived from the ethnobotanical use of plants, bioprospection explores the existing biodiversity through integration of modern omics techniques with targeted bioactivity assays. In this work we set up a bioprospection plan to investigate the phytochemical diversity and the potential bioactivity of five Artemisia species with recognized ethnobotanical tradition (A. absinthium, A. alba, A. annua, A. verlotiorum and A. vulgaris), growing wild in the natural areas of the Verona province. We characterized the specialized metabolomes of the species (including sesquiterpenoids from the artemisinin biosynthesis pathway) through an LC-MS based untargeted approach and, in order to identify potential bioactive metabolites, we correlated their composition with the in vitro antioxidant activity. We propose as potential bioactive compounds several isomers of caffeoyl and feruloyl quinic acid esters (e.g. dicaffeoylquinic acids, feruloylquinic acids and caffeoylferuloylquinic acids), which strongly characterize the most antioxidant species A. verlotiorum and A. annua. Morevoer, in this study we report for the first time the occurrence of sesquiterpenoids from the artemisinin biosynthesis pathway in the species A. alba.

Phytochemical management of root knot nematode (Meloidogyne incognita) kofoid and white chitwood by Artemisia spp. in tomato (Lycopersicon esculentum L.) / Manejo fitoquímico do nematoide do nódulo de raiz (Meloidogyne incognita) kofoid e do chitwood branco por Artemisia spp. em tomate (Lycopersicon esculentum L.)

Abstract In vitro and screen house experiments were conducted to investigate the effectiveness of thirteen phytochemicals from Artemisia elegantissimia and A. incisa on root knot nematode, Meloidogyne incognita in tomato (Lycopersicon esculentum L.) cv. Rio Grande. A positive control (Carbofuran) and negative control (H2O) were also used for comparison. Effectiveness of phytochemicals against juveniles (J2s) mortality and egg hatch inhibition were evaluated after 24, 48 and 72 hours of incubation at three concentrations viz; 0.1, 0.2 and 0.3 mg/mL in vitro conditions. Amongst thirteen phytochemicals, Isoscopletin (Coumarin), Carbofuran and Apigenin (Flavonoid) showed the highest mortality and egg hatch inhibition of M. incognita at all intervals. Inhibition of eggs and J2s mortality were the greatest (90.0%) and (96.0%) at 0.3 mg/mL concentration. Application of phytochemicals caused reduction in number of galls, galling index, and egg masses on tomato plant and enhanced plant growth parameters under screen house conditions. Gall numbers (1.50), galling index (1.00), number of juveniles (4.83) and egg masses (4.00) were greatly reduced and plant growth parameters such as; plant height (28.48 cm), fresh (72.13 g) and dry shoot weights (35.99 g), and root fresh (6.58 g) and dry weights (1.43 g) were increased significantly by using Isoscopletin. In structure activity relationship, juveniles of M. incognita, exhibited variations in their shape and postures upon death when exposed to different concentrations of phytochemicals of Artemisia spp. The present study suggests that Artemisia based phytochemicals possess strong nematicidal effects and can be used effectively in an integrated disease management program against root knot nematodes.

Resumo Experimentos in vitro e de triagem foram conduzidos para investigar a eficácia de treze constituintes fitoquímicos de Artemisia elegantissimia e A. incisa no nematóide de galhas, Meloidogyne incognita em tomateiro (Lycopersicon esculentum L.) cv. Rio Grande. Um controle positivo (carbofuran) e controle negativo (H2O) também foram utilizados para comparação. A eficácia dos fitoquímicos contra a mortalidade juvenil (J2s) e a inibição da eclosão de ovos foram avaliadas após 24, 48 e 72 horas de incubação em três concentrações, tais como: 0,1; 0,2 e 0,3 mg/mL em condições in vitro. Dentre os treze fitoquímicos, isoscopletina (cumarina), carbofurano e apigenina (flavonoide) apresentaram a maior mortalidade e a inibição da eclosão de ovos de M. incognita em todos os intervalos. A inibição da mortalidade dos ovos e J2s foi a maior (90,0%) e (96,0%) na concentração de 0,3 mg/mL. A aplicação de fitoquímicos causou redução no número de galhas, índice de fricção e massa de ovos no tomateiro e melhorou os parâmetros de crescimento das plantas em condições de triagem. Números de galhas (1,50), índice de insetos galhadores (1,00), número de juvenis (4,83) e massas de ovos (4,00) foram bastante reduzidos e os parâmetros de crescimento das plantas, como altura da planta (28,48 cm), peso fresco (72,13 g) e seco (35,99 g), raiz fresca (6,58 g) e peso seco (1,43 g) foram significativamente aumentados usando isoscopletina. Na relação atividade estrutura, juvenis de M. incognita, exibiram variações em sua forma e posturas após a morte quando expostos a diferentes concentrações de fitoquímicos de Artemisia spp. O presente estudo sugere que os fitoquímicos à base de artemísia possuem fortes efeitos nematicidas e podem ser usados ​​eficazmente em um programa integrado de controle de doenças contra nematóides de galhas.

Identification and Quantification of Thujone in a Case of Poisoning Due to Repeated Ingestion of an Infusion of Artemisia Vulgaris L.

Plants of the Artemisia genus are used worldwide as ingredients of botanical preparations. This paper describes the case of a 49-year-old man admitted to the emergency room at a Zurich hospital in a manic state after the ingestion of 1 L of an infusion of Artemisia vulgaris. Two monoterpenic ketones, α- and ß-thujone, are present in various concentrations in Artemisia spp., but adverse effects have previously been associated only with essential oil from Artemisia absinthium and attributed to the inhibition of gamma-aminobutyric acid receptors, with consequent excitation and convulsions. The aim of this work was to examine and quantify the possible presence of thujone in the patient's serum and urine. A High Performance Liquid Chromatography (HPLC) method with isocratic separation and fluorescence detection (FLD) was set up and validated. Serum thujone concentrations were found to be 27.7 ± 3.48 µg/mL at day 0 and 24.1 ± 0.15 µg/mL on day 1. Results were confirmed by a gas chromatography with flame ionization detection (FID). Poisoning due to thujone was thus confirmed, suggesting four possible scenarios: (1) an unusually high concentration of thujone in the A. vulgaris ingested; (2) chronic exposure as the cause of the poisoning; (3) low metabolic efficiency of the patient; (4) contamination or adulteration of the plant material with other Artemisia spp., for example, A. absinthium. PRACTICAL APPLICATION: These results could aid research in the field of adverse effects of botanicals, lead to better understanding and management of similar cases of poisoning, and promote more informed use of natural products.

Bronchial challenge responses in asthmatic patients sensitized to Artemisia spp. pollen

To characterize the patients whose asthma may be caused by Artemisia pollen extracts, we studied the bronchoprovocation test with Korean Artemisia pollen extracts (1:20 w/v), methacholine bronchial challenge test and wormwood-RAST in 32 asthmatic patients sensitized to Artemisia pollen. Twenty-six(81%) developed a 15% or greater decrease in FEVI after the inhalation of Artemisia pollen extracts and 13 patients showed early responses, 8 dual, and 5 late only. Thirteen(50%) out of 26 positive responders complained of seasonal aggravation of their asthmatic symptoms. Seven(53.8%) of the 13 seasonal type patients, 10(76.9%) of the 13 perennial type and 5(100%) of the 5 negative responders showed concurrent positive responses in the house dust bronchoprovocation test. The bronchial responsiveness to allergen(PD15) was more dependent upon the specific IgE level(bound radioactivity on wormwood-RAST) and multiple regression analysis revealed that the specific IgE level and methacholine PC20 may be contributory to allergen PD15. These results suggested that specific IgE to Artemisia pollen appears to be the major contributor to susceptibility to Artemisia bronchial challenges and this pollen may be considered as one of the important allergenic etiologies of atopic asthma in this country.

Progressive Development of Bronchial Asthma from Allergic Rhinitis in a Patient Sensitized to Artemisia SPP. Pollen: A Case Report

The association between allergic rhinitis and bronchial asthma, that is, whether it is a seqential process or independent diseases, has not been established. We confirmed that a 25 year-old woman with allergic rhinitis sensitized to Artemisia spp. pollens and house dust mites developed bronchial asthma during a 2 year follow-up period. Severe broncho-constriction was noted after inhalation of methacholine, and the bronchoprovocation test with extract of Korean Artemisia spp. pollen showed early bronchoconstiction. Total IgE and specific IgE antibodies to Artemisia absinthium increased, compared to the initial levels. It is suggested that some patients with allergic rhinitis progress to bronchial ashtma.