Exploring anti-inflammatory and antioxidant-related quality markers of Artemisia absinthium L. based on spectrum-effect relationship.

INTRODUCTION: Artemisia absinthium L. is a well-known medicinal, aromatic, and edible plant with important medicinal and economic properties and a long history of use in treating liver inflammation and other diseases; however, there has been insufficient progress in quality control. OBJECTIVE: This study aimed to investigate the quality markers for the anti-inflammatory and antioxidant activities of A. absinthium based on spectrum-effect relationship analysis. MATERIALS AND METHODS: Eighteen batches of A. absinthium from different origins were used. Chemical fingerprints were obtained by ultra-performance liquid chromatography (UPLC). The chemical compositions were identified by quadrupole-Orbitrap high-resolution mass spectrometry. Anti-inflammatory activity was assessed by inhibition of cyclooxygenase-2 and 15-lipoxygenase in vitro and inhibition of nitric oxide release in lipopolysaccharide-induced BV-2 cells. Antioxidant activity was assessed by DPPH and ABTS radical scavenging assays. The relationship between bioactivity and chemical fingerprints was then analyzed using chemometrics including gray relational analysis, bivariate correlation analysis, and orthogonal partial least squares analysis. RESULTS: Different batches of A. absinthium extracts possessed significant anti-inflammatory and antioxidant activities to varying degrees. Eighty compounds were identified from A. absinthium, and 12 main common peaks were obtained from the UPLC fingerprints. P3 (chlorogenic acid), P5 (isochlorogenic acid A), and P6 (isochlorogenic acid C) were screened as the most promising active compounds by correlation analysis and further validated for their remarkable anti-inflammatory effects. CONCLUSION: This is the first study to screen the quality markers of A. absinthium by establishing the spectrum-effect relationship, which can provide a reference for the development of quality standards and further research on A. absinthium.

The impact of microcrystalline and nanocrystalline cellulose on the antioxidant phenolic compounds level of the cultured Artemisia absinthium.

Artemisia absinthium has long been used traditionally as an anti-microbial and antioxidant agent. Various biologically active secondary metabolites, including phenolic compounds such as gallic acid and p-coumaric acid, have been reported from the species. In addition, growing the plants under in vitro conditions enriched with elicitors is a cost-effective approach to enhance secondary metabolite production. This paper examined microcrystalline cellulose (MCC) and nanocrystalline cellulose (NCC) effects on morphological characteristics, phenolic compounds, antioxidant activity, and volatile oil content of A. absinthium. The treated shoots with various concentrations of MCC and NCC were subjected to spectrophotometric, GC-MS, and LC-MS analysis. FESEM-EDX, TEM, XRD, and DLS methods were applied to characterize MCC and NCC properties. Morphological findings revealed that the stem length, dry, and fresh weights were improved significantly (P ≤ 0.05) under several MCC and NCC concentrations. Some treatments enhanced gallic and p-coumaric acid levels in the plant. Although 1.5 g/L of MCC treatment showed the highest antioxidant activity, all NCC treatments reduced the antioxidant effect. The findings suggest that both MCC and NCC, at optimized concentrations, could be exploited as elicitors to improve the secondary metabolite production and morphological properties.

Predictive toxicological effects of Artemisia absinthium essential oil on hepatic stellate cells.

Medicinal plants are important worldwide, considering their properties for treating diseases; however, few studies have evaluated their toxicological potential. Among them, Artemisia absinthium is frequently used to treat liver diseases, because its essential oil has several popular therapeutic properties. Based on this information, in the present study, we investigated molecular connectors of physiological effects of the Artemisia absinthium essential oil on human hepatic stellate cell line, LX-2, to explore the potential toxicity of the plant on liver cells. LX-2 is a cellular model to investigate mechanisms of liver fibrosis; then, to analyze the essential oil effects LX-2 was cultured under different conditions, treated or not with the essential oil at 0.4 µg/µL for 24 h. Next, fluorescence microscopy analyses, gene expression measurements, and biochemical approaches revealed that the essential oil reduced pro-fibrogenic markers; however, disrupt lipid metabolism, and cause cellular stress, by the activation of cellular detoxification and pro-inflammatory processes. In conclusion, the hepatic stellate cells incubated with the essential oil present an antifibrotic potential, supporting its popular use; however, the combined results suggest that the essential oil of Artemisia absinthium should be used with caution.

New Evidence for Artemisia absinthium as an Alternative to Classical Antibiotics: Chemical Analysis of Phenolic Compounds, Screening for Antimicrobial Activity.

Artemisia absinthium, an important herb of the Artemisia genus, was evaluated in this study for its potential as an alternative to classical antibiotics. The antimicrobial activity of methanol extracts of A. absinthium (MEAA) was evaluated using the broth microdilution method, revealing that A. absinthium exhibited broad-spectrum antibacterial and antifungal activity. Ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS) was used to analyze the chemical profile of the MEAA, with a focus on flavonoids, quinic acids, and glucaric acids. A total of 90 compounds were identified, 69 of which were described for the first time in A. absinthium. Additionally, a new class of caffeoyl methyl glucaric acids was identified. The main active compounds were quantified and screened for antimicrobial activity. A. absinthium was found to be rich in quinic acids and flavonoids. The screening for antimicrobial activity also revealed that salicylic acid, caffeic acid, casticin, and 3,4-dicaffeoylquinic acid had varying degrees of antimicrobial activity. The acute toxicity of MEAA was examined following OECD guidelines. The administration of 5000 mg/kg bw of MEAA did not result in mortality in male and female mice. Furthermore, there were no observed effects on the visceral organs or general behavior of the mice, demonstrating the good safety of MEAA. This study provides new evidence for the use of A. absinthium as an alternative to classical antibiotics in addressing the problem of bacterial resistance.

Methanolic extract of Artemisia absinthium prompts apoptosis, enhancing expression of Bax/Bcl-2 ratio, cell cycle arrest, caspase-3 activation and mitochondrial membrane potential destruction in human colorectal cancer HCT-116 cells.

The Artemisia absinthium (AA), belongs to the Asteraceae family, is used as a therapeutic agent in traditional medicine in Iran. It is a rich source of biology-active compounds. However, the molecular mechanism of AA contributing to cell proliferation and apoptosis is still unknown. This study aims to assess the anticancer activity of the methanolic extract of A. absinthium (MEAA) against human colorectal cancer HCT-116 cell line. The cytotoxic effects of MEAA on HCT-116 cells was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay. The expression levels of BAX and BCL-2 in HCT-116 cell line were examined by qRT-PCR. Annexin V/PI-flow cytometry technique was used to detect the cell cycle and apoptosis. MMP was predicted by Rhodamine 123 staining, and caspase 3 activity was analyzed by ELISA. Western blot method was performed to detect the expression level of BAX, Bcl-2 and Caspase-3 proteins. The MTT test revealed MEAA reduced the viability of HCT-116 cells. The mRNA and protein levels of BAX increased, but those of BCL-2 decreased in MEAA-treated cells. MEAA also prompted cell cycle arrest and induced apoptosis. After adding MEAA, the protein level and activity of caspase 3 and MMP destruction significantly increased. MEAA predominantly prompted apoptosis in HCT-116 cells by activating the mitochondrial pathway.

Effect of Artemisia absinthium ethanolic extract on oxidative stress markers and the TLR4, S100A4, Bax and Bcl-2 genes expression in the kidney of STZ-induced diabetic rats.

OBJECTIVES: The present study was conducted to examine antidiabetic effects of Artemisia absinthium ethanolic extract [A. absinthium] and to investigate its effects on oxidative stress markers and the expression of TLR4, S100A4, Bax and Bcl-2 genes in the kidney of STZ-induced diabetic rats. METHODS: Thirty six rats (weight 200-250 g) were randomly divided into diabetes and control groups. Induction of diabetes was performed using STZ (55 mg/kg.bw). Biochemical parameters and oxidative stress markers (SOD and MDA) were measured using spectrophotometry after 60 days of treatment. The expression of TLR4, S100A4, Bax and Bcl-2 were analyzed by real-time PCR. One-way analysis of variance (ANOVA) and Bonferroni post hoc test were used to compare the data. RESULTS: Diabetes significantly impairs the serum fasting blood glucose (FBG), lipid profile, urea, creatinine and albumin. At the end of treatment with A. absinthium extract, these parameters were close to the normal range. The results showed that the A. absinthium extract significantly decreased the kidney expression of TLR4, S100A4, Bax and increased the expression of Bcl-2 and improved oxidative stress markers (SOD and MDA) in the kidney tissues of treated rats. Also, all of these beneficial effects of the A. absinthium were dose-dependent. CONCLUSIONS: The extract of A. absinthium possesses antidiabetic effects. A. absinthium decreased the expression of TLR4, S100A4, Bax and increased the expression of Bcl-2 and improved oxidative stress. Therefore, this herbal extract can be used as an adjuvant treatment for diabetic complications.

Bioactive Principles and Potentiality of Hot Methanolic Extract of the Leaves from Artemisia absinthium L "in vitro Cytotoxicity Against Human MCF-7 Breast Cancer Cells, Antibacterial Study and Wound Healing Activity".

BACKGROUND: Artemisia absinthium L is an ornamental plant widespread in Saudi Arabia. Traditionally, the plant has been used in the Arabic medicine. But the scientific evidence of the bioactive compounds and their medicinal value was not yet explored widely. OBJECTIVE: The study was designed to analyse the bioactive principles and medicinal properties of Artemisia absinthium L, a traditional herb grown in southern part of Saudi Arabia. METHODS: The bioactive compounds present in Hot Methanolic Extract of the Leaves (HMEL) of Artemisia absinthium L. was explored by GC-MS analysis. The cytotoxicity effect of HMEL was determined against MCF-7 breast cancer cells ATCC and human colon cancer cells HCT 116 ATCC by performing MTT assay. Morphological changes of HMEL treated MCF-7 were observed under a phasecontrast microscope by staining the cells with neutral red. A Reaction Mixture (RM) of HMEL was prepared in Milli-Q water and antibacterial susceptibility was performed against both Gram-positive and Gram-negative bacteria. Furthermore, in vivo wound healing properties of the RM was screened in male rats and their efficacy was compared with standard povidone iodine cream. Biomarkers such as IL-1ß, IL- 6, TNF- α, caspase-9 and caspase-3 levels were determined to qualify the wound healing property. RESULTS: Epiyangambin, flavone, octadecanoic acid, 2,3-dihydroxypropyl ester, palmitic acid ß - monoglyceride, á-D-mannofuranoside, camphor, and terpineol were identified as possible compounds through GC-MS analysis. The HMEL of Artemisia absinthium L was actively inhibiting the proliferation of breast cancer cells MCF-7 ATCC at the concentration of 80.96 ± 3.94 µg/ml as IC50 value but failed to inhibit the proliferation against the treated human colon cancer cells HCT 116 cells ATCC. HMEL of Artemisia absinthium L was showing a moderate spectrum of antibacterial effect against the screened bacteria. RM showed better wound healing property than standard povidone iodine cream that modulates cytokine networks and apoptosis markers levels indicated the healing of wound. CONCLUSION: The study suggested that novel anticancer, antibacterial and immune modulatory molecules can be developed from the leaves of Artemisia absinthium L.

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.

Multifunctional Hydroxyapatite Coated with Arthemisia absinthium Composites.

There is significant research showing that essential oils extracted from the plants have antibacterial effects. The purpose of this study was to develop a biocomposite based on hydroxyapatite coated with Artemisia absinthium essential oil and to highlight its antibacterial activity. Therefore, present studies are aimed at developing new materials combining hydroxyapatite with Artemisia absinthium essential oil, in order to avoid postoperative infections. The purpose of this work is to highlight the antimicrobial properties of the Artemisia absinthium essential oil-hydroxyapatite composites obtained by a simple method and at low costs. The structural properties and antimicrobial efficiency of the Artemisia absinthium essential oil-hydroxyapatite composite have been studied. The samples based on Artemisia absinthium essential oil analyzed in this study showed that wormwood essential oil presented the highest efficacy against the fungal strain of C. parapsilosis. It has been shown that wormwood essential oil has a strong antimicrobial effect against the microbial strains tested in this study. Furthermore, the antimicrobial properties of the biocomposites based on hydroxyapatite and essential oil are due to the presence of the essential oil in the samples.

Romanian Wormwood (Artemisia absinthium L.): Physicochemical and Nutraceutical Screening.

Artemisia species are used worldwide for their antioxidant, antimicrobial and anti-inflammatory properties. This research was designed to investigate the phytochemical profile of two ethanolic extracts obtained from leaves and stems of A. absinthium L. as well as the biological potential (antioxidant activity, cytotoxic, anti-migratory and anti-inflammatory properties). Both plant materials showed quite similar thermogravimetric, FT-IR phenolic profile (high chlorogenic acid) with mild antioxidant capacity [ascorbic acid (0.02-0.1) > leaves (0.1-2.0) > stem (0.1-2.0)]. Alcoholic extracts from these plant materials showed a cytotoxic effect against A375 (melanoma) and MCF7 (breast adenocarcinoma) and affected less the non-malignant HaCaT cells (human keratinocytes) at 72 h post-stimulation and this same trend was observed in the anti-migratory (A375, MCF7 > HaCat) assay. Lastly, extracts ameliorated the pro-inflammatory effect of TPA (12-o-tetradecanoylphorbol-13-acetate) in mice ears, characterized by a diffuse neutrophil distribution with no exocytosis or micro-abscesses.

The Extracts of Artemisia absinthium L. Suppress the Growth of Hepatocellular Carcinoma Cells through Induction of Apoptosis via Endoplasmic Reticulum Stress and Mitochondrial-Dependent Pathway.

Artemisia absinthium L. has pharmaceutical and medicinal effects such as antimicrobial, antiparasitic, hepatoprotective, and antioxidant activities. Here, we prepared A. absinthium ethanol extract (AAEE) and its subfractions including petroleum ether (AAEE-Pe) and ethyl acetate (AAEE-Ea) and investigated their antitumor effect on human hepatoma BEL-7404 cells and mouse hepatoma H22 cells. The cell viability of hepatoma cells was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The apoptosis, cell cycle, mitochondrial membrane potential (Δψm), and reactive oxygen species (ROS) were analyzed by flow cytometry. The levels of proteins in the cell cycle and apoptotic pathways were detected by Western blot. AAEE, AAEE-Pe, and AAEE-Ea exhibited potent cytotoxicity for both BEL-7404 cells and H22 cells through the induction of cell apoptosis and cell cycle arrest. Moreover, AAEE, AAEE-Pe, and AAEE-Ea significantly reduced Δψm, increased the release of cytochrome c, and promoted the cleavage of caspase-3, caspase-9, and poly(ADP-ribose) polymerase (PARP) in BEL-7404 and H22 cells. AAEE, AAEE-Pe, and AAEE-Ea significantly upregulated the levels of ROS and C/EBP-homologous protein (CHOP). Further, AAEE, AAEE-Pe, and AAEE-Ea significantly inhibited tumor growth in the H22 tumor mouse model and improved the survival of tumor mice without side effects. These results suggest that AAEE, AAEE-Pe, and AAEE-Ea inhibited the growth of hepatoma cells through induction of apoptosis, which might be mediated by the endoplasmic reticulum stress and mitochondrial-dependent pathway.

Shikimic acid from Artemisia absinthium inhibits protein glycation in diabetic rats.

This study investigated the impact of Shikimic Acid (SA) obtained from leaves of Artemisia absinthium on protein glycation in the retina of diabetic rats. The GC/MS analysis of A. absinthium showed that the most abundant bioactive compound was SA (C7H10O5) with a measured retention Index (RI) of 1960 compared to that of the reference sample (1712). Male albino rats were divided into two main groups, Group I (control) and Group II (diabetic); Group II was further divided into four subgroups: Group IIa (diabetic control), Group IIb (diabetic rats were given SA orally [50 mg/kg, body weight (bw)/day], Group IIc diabetic rats were given SA orally [100 mg/kg, bw/day], and Group IId (diabetic rats were given metformin orally [100 mg/kg, bw/day] as positive control). The data obtained suggested that SA reduced glucose and glycated hemoglobin levels. In addition, SA also decreased the formation of glucose-derived advanced glycation end products. Interestingly, SA showed interference with the release of inflammatory mediators in retina and possess antioxidant potential. In conclusion, SA protected the tissues from detrimental effects of hyperglycemia and enhanced antioxidant activity. SA could be a potential lead in the process of drug development in the future to prevent retinopathy in diabetic subjects.

Ecotoxicity of a novel biopesticide from Artemisia absinthium on non-target aquatic organisms.

Biopesticides are increasingly being used to replace synthetic pesticides for pest control. This change raises concern for its environmental impacts, especially on non-target organisms. In this study, the ecotoxicological effects of a potential nematicide from Spanish populations of Artemisia absinthium (var. Candial) were evaluated on freshwater and aquatic non-target organisms. The study focused on the aqueous extract (hydrolate), the principal component of which ((-) -(Z) -2,6-dimethylocta-5,7-diene-2,3-diol) is responsible for its nematicidal effect. Until now, the hydrolate has been considered a byproduct of the process used to obtain essential oils, and there are no studies on its ecotoxicity from any plant with biopesticide properties. Our results indicated that A. absinthium hydrolate caused acute toxicity for non-target organisms at dilutions as low as 0.2%. The sensitivity of the organisms, from the most to the least sensitive, was: Daphnia magna (LC50 = 0,236%) > Vibrio fisheri (LC50 = 1,85%) > Chlamydomonas reinhardtii (LC50 = 16,49). Moreover, the A. absinthium organic extract was highly toxic to D. magna (LC50 = 0,093 mg/L). A. absinthium hydrolate toxicity was also tested on a natural river microbial community. Bacterial growth was not affected; the physiology of the community was only slightly modified, namely through an increased ability to degrade different substrates, mainly carbohydrates. This study provides for the first time an exhaustive assessment of the environmental exposure of a plant-derived biopesticide and shows that these products may cause a broad range of toxicity on non-target aquatic organisms.

Ovicidal and larvicidal activity of extracts from medicinal-plants against Haemonchus contortus.

The use of medicinal plants (MP) containing bioactive compounds is an alternative strategy to control of parasitic nematode of small ruminants Haemonchus contortus at various stages of their life cycle. The aims of this study were to determine the in vitro anthelmintic activity of both aqueous and methanolic extracts from 13 medicinal plants typical for Central Europe, and to determine quantity of selected plant secondary metabolites (PSMs) in the methanolic extracts. In vitro egg hatch test and larval development tests were conducted to determine the possible anthelmintic effects of methanolic and aqueous extracts of the roots of Althaea officinalis L., Petasites hybridus L. and Inula helenium L.; flowers of Malva sylvestris L. and Chamomilla recutita L.; leaves of Plantago lanceolata L. and Rosmarinus officinalis L.; seeds of Foeniculum vulgare Mill. and stems of Solidago virgaurea L., Fumaria officinalis L., Hyssopus officinalis L., Melisa officinalis L. and Artemisia absinthium L. on eggs and larvae of H. contortus. Ultra-performance liquid chromatography and tandem mass spectroscopy was used for quantifying six PSMs: gallic acid (GA), rutin (RU), diosmin (DI), hesperidin (HE), quercetin (QU) and kaempferol (KA). RU content of the most effective methanolic extracts was in the order: M. sylvestris (9.33 mg/g DM) > A. absinthium (6.10 mg/g DM) > C. recutita (0.42 mg/g DM). The highest concentration of QU (44.8 mg/g DM) and KA (6.59 mg/g DM) were detected in stems of F. officinalis comparing to the other evaluated plants. The most significant (p < 0.05) anthelmintic effects exhibited methanolic extracts of A. absinthium in both in vitro tests (i.e., egg hatch test and larval development test). Additionally, only two methanolic extracts of C. recutita and M. sylvestris were comparable to activity of A. absinthium using the larval development test. Wider spectrum of aqueous extracts exhibited stronger ovicidal activity in comparison to methanolic extracts. The similar trend was observed in evaluating of larvicidal activity of aqueous and methanolic plant extracts.

Chicory (Cichorium intybus) and wormwood (Artemisia absinthium) extracts exhibit strong larvicidal activity against mosquito vectors of malaria, dengue fever, and filariasis.

Vector-borne diseases transmitted by mosquitoes cause globally important diseases such as malaria, dengue fever, and filariasis. The incidence of these diseases can be reduced through mosquito control programs but these control programs currently rely on synthetic insecticides that can impact the environment, and has selected widespread mosquito resistance. Environment friendly and biodegradable natural insecticides discovered in plants offer an alternative approach to mosquito control. Here, we investigated extracts from root or aerial parts of Chicory (Cichorium intybus) and wormwood (Artemisia absinthium) against the early 4th instar larvae of Anopheles stephensi (malaria vector), Aedes aegypti (dengue fever vector), and Culex quinquefasciatus (filariasis vector). The root and aerial parts extracts of A. absinthium and C. intybus at 200, 100, 50, 25 and 12.5 ppm caused significant mortality of the tested mosquito species. Root extracts exhibited higher larvicidal activity that aerial part extracts. The highest larvicidal activity was recorded in methanol extract of roots of C. intybus with LC50 = 66.16, 18.88 and LC¬90 = 197.56, 107.16 ppm for An. stephensi; LC50 = 78.51, 40.15 and LC90 = 277.31, 231.28 ppm for Ae. aegypti and LC50 = 103.99, 64.56 and LC¬90 = 314.04, 247.54 ppm for Cx. quinquefasciatus. These results reveal potent mosquito larvicidal activity against vectors of malaria, dengue fever, and filariasis is present in extracts of chicory and wormwood.

Phytochemical screening and antibacterial potential of Artemisia absinthium L., Swertia chirayita and Sphaeranthus indicus.

Utilization of herbs for medicinal purpose started in the early history of mankind several thousand years ago. In this study, some plants that are used for lowering cholesterol level in local areas of Pakistan, such as Artemisia absinthium L., Swertia chirayita and Sphaeranthus indicus were screened for their phytochemical and antibacterial properties. For this purpose, these plants were extracted in different solvents i.e. ethanol, hexane and ethyl acetate. Phytochemcial analysis unveiled the existence of different bioactive compounds in these extracts. Presence of sugars was further confirmed by performing TLC. Antibacterial activity was determined against indicated bacterial strains, among all extracts Gul-e-mundi had maximum inhibition zone (23mm). DPPH free radical assay revealed the significant antioxidative potential of all the extracts where Gul-e-mundi showed maximum potential i.e., 83%. Plant extracts were also showing anti-proliferative activity on root tips of Allium cepa and Gul-e-mundi was observed to have maximum antimitotic activity i.e. 5%. GC-MS analysis revealed that oleic acid and linoleic acid were the compounds responsible for imparting antibacterial potential to Gul-e-mundi. In conclusion, among all the tested extracts Gul-e-mundi had maximum antibacterial, antioxidative and antimitotic potential. For future studies, phytochemcials responsible for these activities can be isolated and modified for pharmacological purpose.

Evaluation of anticancer, antioxidant activity and phenolic compounds of Artemisia absinthium L. Extract.

In the treatment of cancer, which remains a fatal disease, increasingly successful treatment rates of alternative therapies using the power of plants have directed the scientific world towards natural plant resources. This study aimed to examine the anti-cancer and antioxidant properties and identify the phenolic content of the methanolic extract obtained from Artemisia absinthium L. (AR) species, which is used as folk-medicine. The antioxidant activity of the extract was investigated using total phenolics, flavonoids, ABTS and CUPRAC methods. Phenolic component analysis of the plant extract was performed by LC-MS/MS. The anti-cancer property of AR extract was investigated on human colon (DLD-1), endometrium (ECC-1) cancer cells and embryonic kidney (HEK-293) cells. Cytotoxic effects were defined with MTT, apoptotic activity with DNA fragmentation ELISA and AO/EB fluorescent staining, the genotoxic effect with the comet assay and the intracellular oxidative status with TAS and TOS methods. As a result of the study, it was determined that AR extract showed an antioxidant effect, and as a result of the content analysis made with LC-MS/MS, phenolic components were determined, the most abundant being chlorogenic acid, followed by quinic acid, cinnamic acid, rhoifolin and malic acide. AR extract showed cytotoxic activity on DLD-1 and ECC-1 cancer cells, while the cytotoxic effect on HEK-293 cells was determined to be low.  It was determined that by increasing the intracellular amount of free radicals on cancer cells, this led to DNA damage, which consequently led to apoptosis of the cancer cells.

Therapeutic efficacy of Artemisia absinthium against Hymenolepis nana: in vitro and in vivo studies in comparison with the anthelmintic praziquantel.

Hymenolepis nana is a common intestinal tapeworm that affects humans. Drugs are available for the treatment of this infection, including praziquantel (PZQ), nitazoxanide and niclosamide. Although the drug of choice is praziquantel, due to its high cure rates, indicators of the development of PZQ resistance by different parasites have begun to appear over recent decades. Therefore, this study was a trial to find an alternative to PZQ by assessing the activity of the crude aqueous extract of the medicinal herb Artemisia absinthium against H. nana. In vitro, the extract was used against adult worms at concentrations of 1 and 5 mg/ml, in comparison with 1 mg/ml of PZQ. The times of worm paralysis and death were determined. Ultrastructural morphological changes were studied using transmission electron microscopy (TEM). For the in vivo study, infected mice were divided into untreated, PZQ-treated and A. absinthium-treated groups (400 mg/kg and 800 mg/kg). Pre- and post-treatment egg counts per gram of faeces (EPG) were performed; then, the reduction percentages of the EPG and worm burden were calculated. The best results were obtained with praziquantel. Artemisia absinthium induced worm paralysis, death and ultrastructural alterations, such as tegumental damage, lipid accumulation, and destruction of the nephridial canal and the intrauterine eggs, in a dose-dependent manner. Additionally, significant reductions in the EPG and worm burden were recorded in A. absinthium-treated mice. Although the results obtained with A. absinthium were promising and comparable to PZQ, further studies using different extracts, active ingredients and concentrations against different parasites should be conducted.

Absolute configuration of the ocimene monoterpenoids from Artemisia absinthium.

The absolute configuration (AC) of the naturally occurring ocimenes (-)-(3S,5Z)-2,6-dimethyl-2,3-epoxyocta-5,7-diene (1) and (-)-(3S,5Z)-2,6-dimethylocta-5,7-dien-2,3-diol (2), isolated from the essential oils of domesticated specimens of Artemisia absinthium, followed by vibrational circular dichroism (VCD) studies of 1, as well as from the acetonide 3 and the monoacetate 4, both derived from 2, since secondary alcohols are not the best functional groups to be present during VCD studies in solution due to intermolecular associations. The AC follows from comparison of experimental and calculated VCD spectra that were obtained by Density Functional Theory computation at the B3LYP/DGDZVP level of theory. Careful nuclear magnetic resonance (NMR) measurements were compared with literature values, providing for the first time systematic 1 H and 13 C chemical shift data. Regarding homonuclear 1 H coupling constants, after performing a few irradiation experiments that showed the presence of several small long-range interactions, the complete set of coupling constants for 3, which is representative of the four studied molecules, was determined by iterations using the PERCH software. This procedure even allowed assigning the pro-R and pro-S methyl group signals of the two gem-dimethyl groups present in 3.

Toxic essential oils. Part V: Behaviour modulating and toxic properties of thujones and thujone-containing essential oils of Salvia officinalis L., Artemisia absinthium L., Thuja occidentalis L. and Tanacetum vulgare L.

Neurotoxic thujones (α- and ß-diastereoisomers) are common constituents of plant essential oils. In this study, we employed a statistical approach to determine the contribution of thujones to the overall observed behaviour-modulating and toxic effects of essential oils (Salvia officinalis L., Artemisia absinthium L., Thuja occidentalis L. and Tanacetum vulgare L.) containing these monoterpene ketones. The data from three in vivo neuropharmacological tests on rats (open field, light-dark, and diazepam-induced sleep), and toxicity assays (brine shrimp, and antimicrobial activity against a panel of microorganisms), together with the data from detailed chemical analyses, were subjected to a multivariate statistical treatment to reveal the possible correlation(s) between the content of essential-oil constituents and the observed effects. The results strongly imply that the toxic and behaviour-modulating activity of the oils (hundreds of constituents) should not be associated exclusively with thujones. The statistical analyses pinpointed to a number of essential-oil constituents other than thujones that demonstrated a clear correlation with either the toxicity, antimicrobial effect or the activity on CNS. Thus, in addition to the thujone content, the amount and toxicity of other constituents should be taken into consideration when making risk assessment and determining the regulatory status of plants in food and medicines.