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Plant-derived natural compounds are desired for their anti-inflammatory properties. We investigated the chemical composition and the anti-inflammatory effects of the essential oil obtained from leaves of the Chinese medicinal plant Artemisia annua L. Artemisia annua L. essential oil (AO) was obtained by hydrodistillation; the extraction yield was approximately 1.2%. Gas chromatography–mass spectrometry (GC–MS) revealed the presence of 13 compounds in AO; the major compounds were caryophyllene (32.16%), eucalyptol (28.62%), α-pinene (19.08%), and β-pinene (14.72%). Cell counting kit-8 (CCK8) assay revealed that AO did not show in vitro cytotoxicity toward RAW264.7 cells and suppressed the expression of nitric oxide (NO), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6) in lipopolysaccharide (LPS)-induced RAW264.7 cells. Pretreatment with AO significantly reduced xylene-induced inflammation in the ears of Kunming mice. AO inhibited inflammatory cell infiltration, decreased epidermis thickness in the ear, and suppressed expression of TNF-α in the serum. The in vivo anti-inflammatory activity of AO was similar to that of the positive control compound Di Song Zhang Bo Cream. Overall, AO was observed to be an effective anti-inflammatory agent. These results provide an experimental basis for commercializing AO-derived external preparations for pharmacological and cosmetic industries.

For the development of genetic programs with novel characteristics induced mutagenesis has been used extensively. Chlorophyll mutations are considered as the most dependable indices for assessing the efficiency of different mutagens in inducing the genetic variability in crop plants and are also used as genetic markers in basic and applied research. In the present scenario of high health susceptibility, the global demand for natural medicine derived from plant species has increased enormously. Sweet wormwood (Artemisia annuaL.) – an important medicinal plant species with immense remedial values, was selected for the present study and exposed to gamma rays at 100 Gy, 200 Gy, 300 Gy and combination treatments with 100 Gy + 0.1%EMS, 200 Gy + 0.1% EMS, 300 Gy + 0.1%EMS. Meiotic study was also done and various cytological aberrations were observed in M2 generation like stickiness, precocious, scattering, laggard and bridge etc. The frequency of induced chl mutation varied in different mutagen treatments. Eight different types of chl mutants namely albina, chlorina, xantha, aurea, viridis, yellow viridis and tigrina etc. were recorded in M2 generation on plant population basis. The frequency of xantha mutants was quite high in both the treatments but in gamma exposed set it was followed by albina whereas in combination treatments viridis was second highest mutant. In different mutants quantitative analysis of chl pigments was also done and content was highest in viridis i.e. 3.86µg/ml FW and lowest in albina i.e. 0 µg/ml FW . Although chl mutations thought to be lethal in nature, but present study has proven to be a milestone in identifying the threshold dose of a mutagen that would increase the genetic variability and induces new trait.

The highly diverse nature of plant microbiota is indicative of many ecological roles of this group of microorganisms. For important medicinal plants such as Artemisia annua L (Family Asteraceae), there can be numerous functional attributes of the associated bacterial endophytes, but the diversity and functions of such endophytes remain widely unknown. To understand their potential functional diversity, we isolated endophytic bacteria associated with the plant from different plant parts including leaves, stems, and roots. Bacterial community of the leaves showed more diversity than those of the stems and the roots. Bacterial isolates were screened in vitro for various plant growth promoting attributes including Indole acetic acid (IAA), 1-aminocyclopropane-1-carboxylate (ACC) deaminase production, and nitrogen fixation. Ninety five percent of the isolates were able to produce IAA, while 7.5% produced ACC deaminase, and 15% were able to fix nitrogen. Based on their potential plant growth promoting attributes, 25 bacterial isolates of 16 genera belonging to four phyla were identified. Proteobacteria (36%) and Firmicutes (36%) were the most predominant isolated bacteria, followed by Actinobacteria (24%) and Bacteroidetes (4%). The most promising isolates were selected for the investigation of their ability to enhance seed germination. We observed that the treatment with isolates AAR80 Bacillus selenatarsenatis and AAL56 Achromobacter xylosoxidans increased germination percentage of inoculated seeds. Moreover, (48%) of the isolates showed antimicrobial activity upon testing against indicator microorganisms. This study revealed that endophytic bacteria isolated from A. annua L have dual potential to be used as growth promoters and antimicrobial agents. [Display omitted] • Isolated 25 endophytic bacteria from Artemisia annua enhanced plant growth via IAA, ACC, production and nitrogen fixation. • The 25 isolates were characterized to be from 16 different genera belonging to 4 phyla. • Proteobacteria (36%) and Firmicutes (36%) were the most predominant isolated bacteria. • Two indigenous bacterial endophytes enhanced A. annua seed germination in pot trials. • Eleven strains showed promising antimicrobial activities, with phyla actinobacteria showing the highest activity. [ABSTRACT FROM AUTHOR]

ABSTRACT Artemisia annua L. is a widely distributed medicinal plant and well-known for treating malaria due to the artemisinin content. We previously found enhanced antioxidant and antibacterial activities of Lactobacillus plantarum-fermented A. annua dried leaves in vitro. The present study compared the effects of the dietary supplementation of L. plantarum, fermented (FA) or non-fermented (NFA) A. annua on laying performance, egg quality, serum cholesterol, and egg yolk oxidative stability during storage in 40-weeks-old Hy-Line Brown layers. In total, 180 layers were randomly allocated into four treatments for 6 weeks: basal diet (CON), basal diet + 0.5% L. plantarum only (LO), basal diet + 0.5% NFA, and basal diet + 0.5% FA. Each treatment comprised five replicates with nine birds each. Egg weight of NFA and FA groups were significantly higher as compared with the CON and LO groups (p

Plants are a potential source of active molecules and are environmentally safer and cheaper than synthetic antibiotics. Bioactive compounds of Artemisia annua have shown pharmacological activities and are used globally as a supplement. The present study tested whether dietary supplementation with alcohol extract of the plant A. annua (ae-Aa; patent BR10201902707) improves the health status of juvenile Nile tilapia and increases resistance to diseases when fish are challenged with the bacteria Aeromonas hydrophila. The experimental design was completely randomized with four experimental groups (0.0, 0.1, 0.25, and 0.5% ae-Aa in the diets) with five repetitions (12 fish per repetition/experimental unit). We assessed serum glucose and cortisol levels in plasma, leukocyte respiratory activity, total plasma protein, serum lysozyme levels, as well as the number of circulating red blood cells and fish leukocytes at the end of the 30 days of feeding (phase I) and 24h after exposure to bacteria (phase II). The levels of lipid peroxidation, catalase activity and glutathione S-transferase of fish were also analyzed. The supplementation of 0.5% of ae-Aa was sufficient to increase the respiratory burst of leukocyte and lysozyme activity, total plasma protein, blood thrombocytes, neutrophils and monocytes after bacterial challenge (P < 0.05), and minimized stress response with decreases in plasmatic glucose and cortisol, and reduction in lipid peroxidation levels (P < 0.05). Results of the present study suggest that ae-Aa as a dietary supplement has benefits, including supplementation with 0.5% A. annua extract for 30 days to minimize the stress response and modulate innate immunity in Nile tilapia, providing fish with greater resistance and disease protection. • Previous dietary supplementation of Artemisia annua alcohol extract enhanced innate immunity of the Nile tilapia. • Feed supplemented with Artemisia annua alcohol extract minimized stress response of the Nile tilapia. • The A. annua alcoholic oral suplementation increased the resistance of the fish challenged with A. hydrophila. [ABSTRACT FROM AUTHOR]

• Polar fraction of A. annua demonstrated remarkable hepatoprotective, antihyperglycemic and antioxidant activities in animal models. • Five quinic acid derivatives; were isolated for the first time from the polar fraction. • The content of quinic acid derivatives in A. annua leaves was determined by HPLC using 3-feruloylquinic acid as marker substance. • Seasonal variation of quinic acid derivatives using HPLC showed that the early summer leaves have the highest content. Genus Artemisia is widely known to have various therapeutic applications and chemical constituents. A. annua L. (commonly known as sweet wormwood or Qinghao) is an annual herb native to China, traditionally used for treating fever and malaria, and as a source of artemisinin. In the present study, a bio-guided fractionation of the 70% ethanolic extract of the leaves of A. annua cultivated in Egypt produced a bioactive polar fraction. Daily doses of this fraction (100 mg/kg b.wt) for 4 weeks substantially reduced the level of the CCl 4 -mediated increase in the liver enzymes; AST, ALT and ALP. Similarly, daily doses of the polar fraction (100 mg/kg) significantly reduced the blood glucose level in alloxan-induced diabetic by 32.1% in the second week and 46.9% in the fourth week, relative to that demonstrated by metformin (66.2%), and significantly (p < 0.01) restored to normal the blood glutathione level (35.2 ± 1.3 mg/dL), almost identical to that shown by vitamin E. Further purification of the bioactive fraction led to the isolation of 5 quinic acid derivatives; 3-feruloylquinic acid, 3,5-dicaffoeylquinic acid, 4,5-dicaffoeylquinic acid, 3,4-dicaffoeylquinic acid, and 3,4-dicaffoeylquinic acid methyl ester. The isolated compounds are reported here for the first time in A. annua , leaves cultivated in Egypt and suggested to be responsible, at least in part, to the biological activities of the polar fraction. Seasonal variation in the content of quinic acid derivatives in the leaves of A. annua L. was investigated during four harvest seasons (March, May, July "pre-flowering stage" and August-September "flowering stage") using RP-HPLC. The content of 3-feruloylquinic acid varied greatly throughout the year. It was found to be the lowest (0.036% w/w) in March "leaf stage" but increased through the warmer months from May to June (0.61% w/w), and reached the highest (1.34% w/w) in leaves harvested in July "early summer". Then-after, gradual decline in the content of quinic acid derivatives was evident in leaves collected during flowering stage in late summer "August-September" (0.84% w/w). [ABSTRACT FROM AUTHOR]

The study was designed to investigate the antibacterial effect of Helicobacter pylori against 32 medicinal plants commonly used as health foods. The medicinal plants used in this study were 32 kinds of medicinal plant extracts using the disk diffusion method for H. pylori activity, which can be eaten every day by everyone. Amoxicillin sodium (150 mg / ml, Ildong Pharmaceutical) and Metronidazole 50 mg / ml) was used as a control group. We measured the area of the transparent area and evaluated that the larger the area, the more effective it is for H. pylori. As a result of this study, the clear zone of inhibition was highest at 372.90mm2, second was 358.30mm2, and Chungho was 348.32mm2. The positive control group, Metronidazole (50 mg / ml CJ), was 503,29 mm2. In the future, the development of antimicrobial materials of various medicinal plants is expected to be effective for the inhibition of H. pylori. [ABSTRACT FROM AUTHOR]

Artemisinin, which is extracted from the plant Artemisia annua L., is a crucial drug for curing malaria and has potential applications for treating cancer, diabetes, pulmonary tuberculosis, and other conditions. Demand for artemisinin is therefore high, and enhancing its yield is important. Artemisinin dynamics change during the growth cycle of A. annua ; however, the regulatory networks underlying these changes are poorly understood. Here, we collected A. annua leaves at different growth stages and identified target genes from transcriptome data. We determined that WRKY6 binds to the promoters of the artemisinin biosynthesis gene artemisinic aldehyde Δ 11(13) reductase (DBR2). In agreement, overexpression of WRKY6 in A. annua resulted in higher expression levels of genes in the artemisinin biosynthesis pathway and greater artemisinin contents than in the wild type. When expression of WRKY 6 was down-regulated, artemisinin biosynthesis pathway genes were also down-regulated and the content of artemisinin was lower. WRKY6 mediates the transcriptional activation of artemisinin biosynthesis by binding to the promoter of DBR2 , making it a key regulator for modulating the dynamics of artemisinin changes during the A. annua growth cycle. • AaWRKY6 reveals artemisinin accumulation during growth in Artemisia annua. • AaWRKY6 enhances the artemisinin content by binding to the promoter of DBR2. • Expression patterns of AaWRKY6 are similar to DBR2. [ABSTRACT FROM AUTHOR]

The anti-malarial drug artemisinin is extracted from the leaves of Artemisia annua L. Due to toxicity to some microorganisms, the release of artemisinin from this medicinal plant in commercial cultivation might produce a potential risk for phosphorus (P) solubilizing bacteria (PSB). Therefore, the growth, P mobilization, and proton and organic acid efflux by two PSB isolates, Bacillus subtilis and Pseudomonas fluorescens , obtained from the soil without growing A. annua L. in history in the region for growing A. annua L., Chongqing, China, were studied through soil and solution incubations with different nominal concentrations of artemisinin (0, 2.5, 5.0, and 10.0 mg/kg or mg/L). Addition of artemisinin into soil and culture solutions decreased significantly the number of PSB except P. fluorescens at a low artemisinin concentration (2.5 mg/L) in culture solution which remained unchanged in comparison with the control (without artemisinin). This suggests high artemisinin inhibited the cell division or led to the death of PSB, and the different species responded differently to artemisinin. Compared with original soil, PSB inoculation significantly increased Olsen P, whilst the addition of artemisinin decreased this P form in soil. There was a positive correlation between the number of PSB and Olsen P content in soils ( r 2 = 0.824, n = 8), indicating the involvement of PSB in P mobilization of insoluble minerals. Oxalate and acetate were commonly found in the bacterial culture solutions, which accounted for 73.6–84.4% of all organic acids in the culture medium without artemisinin. Malate was detected in the culture solution of B. subtilis , and citrate and succinate in P. fluorescens . The percentage of tricalcium phosphate solubilization (PTPS) positively correlated to the concentrations of protons and all organic acids ( r 2 proton ＝0.901, n＝8, P ＜0.01; r 2 organic acids ＝0.923, n＝8, P ＜0.01). The concentrations of protons, organic acids and soluble inorganic P in culture solutions, and PTPS were decreased simultaneously as nominal artemisinin concentrations increased. For these decreases it implies the metabolic inhibition and the death of PSB caused by artemisinin could be the main reasons for the less efflux of protons and organic acids, presumably resulting in the decreased ability of PSB to mobilize inorganic P. Therefore, artemisinin released from A. annua L. in commercial and continual cultivation could adversely affect the community structure and inorganic P mobilization of PSB in soils. [ABSTRACT FROM AUTHOR]

Currently, the most effective antimalarial is artemisinin, which is extracted from the leaves of medicinal herb Artemisia annua L. (A. annua). Previous studies showed that the complex chemical matrix of A. annua could enhance both the bioavailability and efficacy of artemisinin. The present study aims to evaluate the efficacy and pharmacokinetic properties of a combination therapy based on artemisinin and 3 components from A. annua with high content (arteannuin B, arteannuic acid, and scopoletin). In vivo antimalarial activity was assessed following a 4-day treatment in murine malaria models (Plasmodium yoelii and Plasmodium berghei). Results showed that a much sharper reduction in parasitemia (~93%) was found in combination therapy compared with pure artemisinin (~31%), indicating pharmacodynamic synergism occurring between artemisinin and arteannuin B, arteannuic acid, and scopoletin. Multiple-dose pharmacokinetics further demonstrated that combination therapy results in increased area under the curve (AUC0→∞ ), Cmax , and t1/2 by 3.78-, 3.47-, and 1.13-fold in healthy mice, respectively, and by 2.62-, 1.82-, and 1.22-fold in P. yoelii-infected mice, respectively. The calculated oral clearance of combination therapy in healthy and P. yoelii-infected mice was also reduced. These findings imply that specific components in A. annua might offer a possibility to develop new artemisinin-based natural combination therapy for malaria treatment. [ABSTRACT FROM AUTHOR]

The anti-malarial drug artemisinin is extracted from the leaves of Artemisia annua L. The potential risk of artemisinin released from leaf debris and root exudation of this medicinal plant is unknown for microbes in soils where commercial cultivation of A. annua L. is practiced. Thus, a soil incubation experiment was conducted to compare microbial biomass, enzyme activities, and bacterial community compositions by different concentrations of artemisinin (without artemisinin, CK; 10 mg kg － 1 soil, LA; and 20 mg kg － 1 soil, HA). The results showed that this compound decreased significantly soil microbial biomass carbon and nitrogen, and activities of dehydrogenases and urease, implying the inhibition on the growth, reproduction, and metabolism of some bacteria, taking into account of the facts that the microbial biomass includes those of bacteria and dehydrogenases cannot act on their own without viable bacterial hosts. There were 13064, 11738, and 10107 16S rDNA sequences picked up from soils of CK, LA, and HA, respectively, by MiSeq Illumina sequencing, which were attributed into 541 (CK), 453 (LA), 414 bacterial phylotypes (HA). Soils received artemisinin showed lower bacterial richness and diversity indexes than CK. Less group and number of bacteria present in soils could encourage over reproduction of any single bacterial taxon and thus increase the risk of plant disease occurrence. All soils studied were dominated by three bacterial phyla (Acidobacteria, Proteobacteria, and Actinobacteria), with these three phyla accounting for 60.32-70.43% of the total bacteria. However, most of bacterial phyla and predominant phylotypes varied significantly in their abundances between CK and artemisinin treatment. Among the top 20 predominant bacteria, five to seven were unique in each of soils studied. Therefore, artemisinin released from A. annua L. in cultivation might exhibit selective antibacterial action towards soil bacteria and cause great changes in bacterial activities and community structure, which could influence, at least in part, soil processes and functions. [ABSTRACT FROM AUTHOR]

In this paper, the extraction of essential oils fromArtemisia annuaL. with supercritical carbon dioxide method has been studied by means of the parameter optimization using response surface methodology. The extracted oil was then purified with molecular distillation. The results showed that the response surface methodology was an effective tool to predict optimal values of extraction parameters, with which the crude oil yield of 11.17% could be achieved at 25 MPa pressure, 50°C, 25 g/minute carbon dioxide flow rate and 2 hours extraction time. The purification below 120°C could produce high-quality essential oil with the yield of 56.1%. The essential oil was mainly composed of six bioactive terpenes, namely limonene, (1s)-α-pinene, β-pinene, β-farnesene, α-caryophyllene, γ-elemene. It was suggested thatA. annuaL. was a potential source of antimicrobial/ antioxidant compounds and the combination of supercritical carbon dioxide and molecular distillation was a good process for the isolation of essential oil. [ABSTRACT FROM PUBLISHER]

Dihydroartemisinic acid (DHAA) is the direct precursor to artemisinin, an effective anti-malaria compound from Artemisia annua L. ( A. annua), and it can be transformed to artemisinin without the catalysis of enzyme. A rapid and sensitive analysis of DHAA in A. annua is needed to screen excellent plant resources aimed to improve artemisinin production. In order to develop a rapid and sensitive determination method for DHAA in plant, the extraction and analysis conditions were extensively investigated in the present work. As a result, extraction of powdered A. annua leaves at 55°C for 50 min with chloroform resulted in the highest yield of DHAA, with a recovery of >98%. The precision of this gas chromatographic procedure ranged from 1.22 to 2.94% for intra-day and from 1.69 to 4.31% for inter-day, respectively. The accuracy was 99.55-103.02% for intra-day and 98.86-99.98% for inter-day, respectively. The measured LOQ and LOD values of the proposed method reached 5.00 and 2.00 μg/mL, respectively. Validation indicated the method was robust, quick, sensitive and adequate for DHAA analysis. [ABSTRACT FROM AUTHOR]

Artemisinin, a sesquiterpene lactone isolated from Artemisia annua L. (sweet wormwood), is extensively used in the treatment of malaria. In order to better understand the metabolism of terpenes in A. annua and the influence of terpene synthases on artemisinin yield, the expression pattern of a monoterpene alcohol dehydrogenase (ADH2) has been studied using transgenic plants expressing promoter-β-glucuronidase ( GUS) fusion. ADH2 played a major role in monoterpenoid biosynthesis including carveol, borneol, and artemisia ketone through in vitro biochemical analysis. In this study, the ADH2 promoter was cloned by the genome walking method. A number of putative cis-acting elements were predicted in promoter region, suggesting that the ADH2 is driven by a complex regulation mechanism. ADH2 gene was highly expressed in old leaves, whereas the artemisinin biosynthetic genes were mainly expressed in bud and young leaves. The expression of ADH2 gene increased quickly during leaf development, revealed by qRT-PCR. GUS expression analysis in different tissues of transgenic A. annua demonstrates that ADH2 expression is exclusively located to T-shaped trichome, not glandular secretory trichome. [ABSTRACT FROM AUTHOR]

In this study, we evaluate the different biological activities of Artemisia annua L., locally known as "Moureh", in various altitudes in North of Iran, which has been used as sedative, fever few, anti inflammation, insecticide and anti infection to treat many current diseases. Parts of plants were collected from two different localities (23-1000 m) in Mazandaran province, North of Iran. The most important of secondary metabolites of total phenolics (TP), total flavonoids (TF) and total anthocyanin (TA) content of extracts were investigated by spectrophotometry method and their antioxidant activity were obtained by Total Antioxidant Capacity (TAC), Reducing Power (RP) and 1,1-diphenyl-2-picryl hydrazyl radical scavenging (DPPH). The essential oils were obtained by hydro distillated in a Clevenger-type apparatus for 5h and analyzed by GC/MS. Results indicate that the main components in essential oils were identified as Artemisia ketone (25.54- 13.6%), followed by 1,8-cineole (11.98-13.26%), camphor (11.89-13.68%), α-pinene (10.11-9.29%) in AFRACHAL (1000m) and DOLAT ABAD (23m) regions , respectively. TP content had significant variation in different plant parts and regions, ranging from (11.22 to 16.94) mgGAEg-1, TF content (11.62 to 63.74) mgQUE g-1and quantity of TA (0.03 to 3.59) mgCGEg-1.The highest contents of secondary metabolites were found in aerial parts when compared with the other parts. Amount of antioxidant activity(IC50) in various parts of A. annua L. was measured (1.98 to 4.2) in DPPH, (7.07 to 7.46) in TAC and (5.26 to 8.04) in RP methods. In general, the highest contents of activities were found in aerial parts when compared with the other parts, whereas this part with the highest amount of IC50 had the weakest antioxidant activity.

Hydrodistilled volatile oils fromAchillea millefoliumL.,Arnica montanaL.,Artemisia absinthiumL., andArtemisia annuaL. were analyzed by GC/MS. One hundred sixty-seven compounds were identified in all samples, ranging from 74.3% to 96.4% of the total oil. A high percentage of oxygenated monoterpenes is the main characteristic ofA. millefoliumandA. absinthiumoils with camphor (19.2%) and isoascaridol (21.9%) as the major constituents, respectively. In contrast, the main component ofA. annuaoil was oxygenated sesquiterpene selina-3,11-dien-6-α-ol (9.6%), while the chemical composition ofA. montanaoil was characterized by a high content of fatty acids withn-hexadecanoic acid (16.1%) as the main constituent. Antioxidant activity was tested using four different methods, DPPH, ABTS, Reducing power, and Phosphomolybdenum assay. The highest antioxidant activity had essential oil ofA. montanausing DPPH and Phosphomolibdenum assay,A. millefoliumin ABTS method, whileA. absinthiumoil showed the best ability to reduce Fe ions. [ABSTRACT FROM AUTHOR]

Phenylalanine ammonia-lyase (PAL) is the key enzyme in the biosynthetic pathway of salicylic acid (SA). In this study, a full-length cDNA of PAL gene (named as AaPAL1) was cloned from Artemisia annua. The gene contains an open reading frame of 2,151 bps encoding 716 amino acids. Comparative and bioinformatics analysis revealed that the polypeptide protein of AaPAL1 was highly homologous to PALs from other plant species. Southern blot analysis revealed that it belonged to a gene family with three members. Quantitative RT-PCR analysis of various tissues of A. annua showed that AaPAL1 transcript levels were highest in the young leaves. A 1160-bp promoter region was also isolated resulting in identification of distinct cis-regulatory elements including W-box, TGACG-motif, and TC-rich repeats. Quantitative RT-PCR indicated that AaPAL1 was upregulated by salinity, drought, wounding, and SA stresses, which were corroborated positively with the identified cis-elements within the promoter region. AaPAL1 was successfully expressed in Escherichia. coli and the enzyme activity of the purified AaPAL1 was approximately 287.2 U/mg. These results substantiated the involvement of AaPAL1 in the phenylalanine pathway. [ABSTRACT FROM AUTHOR]

Recent evidence suggests that polyphenolic compounds from plants have anti-invasion and anti-metastasis capabilities. The Korean annual weed, Artemisia annua L., has been used as a folk medicine for treatment of various diseases. Here, we isolated and characterized polyphenols from Korean A. annua L (pKAL). We investigated anti-metastatic effects of pKAL on the highly metastatic MDA-MB-231 breast cancer cells especially focusing on cancer cell adhesion to the endothelial cell and epithelial-mesenchymal transition (EMT). Firstly, pKAL inhibited cell viability of MDA-MB-231 cells in a dose-dependent manner, but not that of human umbilical vein endothelial cells (ECs). Polyphenols from Korean A. annua L inhibited the adhesion of MDA-MB-231 cells to ECs through reducing vascular cell adhesion molecule-1 expression of MDA-MB-231 and ECs, but not intracellular adhesion molecule-1 at the concentrations where pKAL did not influence the cell viability of either MDA-MB-231 cells nor EC. Further, pKAL inhibited tumor necrosis factor-activated MDA-MB-231 breast cancer cell invasion through inhibition of matrix metalloproteinase-2 and matrix metalloproteinase-9 and EMT. Moreover, pKAL inhibited phosphorylation of Akt, but not that of protein kinase C. These results suggest that pKAL may serve as a therapeutic agent against cancer metastasis at least in part by inhibiting the cancer cell adhesion to ECs through suppression of vascular cell adhesion molecule-1 and invasion through suppression of EMT. Copyright © 2016 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

The treatment of acanthamoebiasis is a great problem. Most cerebral invasions end with death, and the treatment of ocular invasions is usually long-lasting and not very effective. Numerous plant extracts and substances isolated from plants, which are effective against trophozoites or cysts, have been studied in the treatment of acanthamoebiasis. However, no agents that are simultaneously effective against both developing forms of amoebae have been discovered yet. It seems that such a plant which fulfils both tasks is Artemisia annua L. Our studies showed that water, alcohol and chloroform extracts from the herb A. annua L. can be applied in general and local treatment or in combined therapy with antibiotics in the treatment of acanthamoebiasis. Extracts from this plant show not only in vitro but also in vivo effects. Studies carried out on experimental animals infected with amoebae show that the application of these extracts significantly prolongs the survival of the animals. [ABSTRACT FROM AUTHOR]

An annual Korean weed, Artemisia annua L., has been used as a folk medicine for the treatment of a number of diseases. Remarkably, among the 32 polyphenols characterized in various parts of plant tissue, including flowers, leafs, stems and roots, 10 compounds were detected for the first time using liquid chromatography-tandem mass spectrometry (LC/MS/MS). The quantification method was validated using structurally related external standards with determination coefficients ( R2) ≥0.9995. The limits of detection and quantitation were 0.068-3.932 and 0.226-13.108 mg/L, respectively. The recoveries estimated at 50 and 100 mg/L ranged between 60.6-92.2 and 61.3-111%, respectively, with relative standard deviations <12%. The roots contained the largest concentration of identified components, while the flowers contained the least. The antioxidant capacity evaluated in terms of 1,1-diphenyl-2-picrylhydrazyl and 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation-scavenging activities and reducing power was highest in the roots and lowest in the flowers. The findings are well correlated and suggest that the antioxidant capacities principally depend upon the polyphenol concentrations in each part of the plant. Copyright © 2015 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

There are many biosynthetic pathways competing for the metabolic flux with the artemisinin biosynthetic pathway in Artemisia annua L. To study the relationship between genes encoding enzymes at branching points and the artemisinin biosynthetic pathway, β-caryophyllene, β-farnesene and squalene were sprayed on young seedlings of A. annua. Transient expression assays indicated that the transcription levels of β-caryophyllene synthase (CPS), β-farnesene synthase (BFS) and squalene synthase (SQS) were inhibited by β-caryophyllene, β-farnesene and squalene, respectively, while expression of some artemisinin biosynthetic pathway genes increased. Thus, inhibition of these genes encoding enzymes at branching points may be helpful to improve the artemisinin content. For further study, the expression levels of four branch pathway genes CPS, BFS, germacrene A synthase (GAS) and SQS were down-regulated by the antisense method in A. annua. In anti-CPS transgenic plants, mRNA levels of BFS and ADS were increased, and the contents of β-farnesene, artemisinin and dihydroartemisinic acid (DHAA) were increased by 212, 77 and 132%, respectively. The expression levels of CPS, SQS, GAS, amorpha-4,11- diene synthase (ADS), amorphadiene 12-hydroxylase (CYP71AV1) and aldehyde dehydrogenase 1 (ALDH1) were increased in anti-BFS transgenic plants and, at the same time, the contents of artemisinin and DHAA were increased by 77% and 54%, respectively, and the content of squalene was increased by 235%. In anti-GAS transgenic plants, mRNA levels of CPS, BFS, ADS and ALDH1 were increased. The contents of artemisinin and DHAA were enhanced by 103% and 130%, respectively. In anti-SQS transgenic plants, the transcription levels of BFS, GAS, CPS, ADS, CYP71AV1 and ALDH1 were all increased. Contents of artemisinin and DHAA were enhanced by 71% and 223%, respectively, while β-farnesene was raised to 123%. The mRNA level of artemisinic aldehyde 11(13) reductase (DBR2) had changed little in almost all transgenic plants. [ABSTRACT FROM AUTHOR]

Application of traditional Chinese drug, artemisinin, originally derived from Artemisia annua L., in malaria therapy has now been globally accepted. Artemisinin and its derivatives, with their established safety records, form the first line of malaria treatment via artemisinin combination therapies (ACTs). In addition to its antimalarial effects, artemisinin has recently been evaluated in terms of its antitumour, antibacterial, antiviral, antileishmanial, antischistosomiatic, herbicidal and other properties. However, low levels of artemisinin in plants have emerged various conventional, transgenic and nontransgenic approaches for enhanced production of the drug. According to WHO (), approximately 3.2 billion people are at risk of this disease. However, unfortunately, artemisinin availability is still facing its short supply. To fulfil artemisinin's global demand, no single method alone is reliable, and there is a need to collectively use conventional and advanced approaches for its higher production. Further, it is the unique structure of artemisinin that makes it a potential drug not only against malaria but to other diseases as well. Execution of its action through multiple mechanisms is probably the reason behind its wide spectrum of action. Unfortunately, due to clues for developing artemisinin resistance in malaria parasites, it has become desirable to explore all possible modes of action of artemisinin so that new generation antimalarial drugs can be developed in future. The present review provides a comprehensive updates on artemisinin modes of action and strategies for enhanced artemisinin production at global level. [ABSTRACT FROM AUTHOR]

Malaria is still one of the major diseases in the world, causing physical and economic problems in tropical regions. Artemisinin (Qinghaosu), a natural compound identified in Artemisia annua L. , is an effective drug mainly against cerebral malaria. The action of this drug is immediate and parasitaemia in the treatment of drug-resistant malaria is rapidily reduced, justifying the industrial production of artemisinin. This article focuses on the industrial production of this potent antimalarial drug, including strategies for enhancing yield using inexpensive and easy steps.

Artemisinin (AN) and artemisinic acid (AA), valuable phyto-pharmaceutical molecules, are well known anti-malarials, but their activities against diseases like cancer, schistosomiasis, HIV, hepatitis-B and leishmaniasis are also being reported. For the simultaneous estimation of AN and AA in the callus and leaf extracts of A. annua L. plants, we embarked upon a simple, rapid, selective, reliable and fairly economical high performance thin layer chromatography (HPTLC) method. Experimental conditions such as band size, chamber saturation time, migration of solvent front and slit width were critically studied and the optimum conditions were selected. The separations were achieved using toluene-ethyl acetate, 9:1 (v/v) as mobile phase on pre-coated silica gel plates, G 60F254. Good resolution was achieved with Rf values of 0.35 ± 0.02 and 0.26 ± 0.02 at 536 nm for AN and 626 nm for AA, respectively, in absorption-reflectance mode. The method displayed a linear relationship with r2 value 0.992 and 0.994 for AN and AA, respectively, in the concentration range of 300-1500 ng for AN and 200-1000 ng for AA. The method was validated for specificity by obtaining in-situ UV overlay spectra and sensitivity by estimating limit of detection (30 ng for AN and 15 ng for AA) and limit of quantitation (80 ng for AN and 45 ng for AA) values. The accuracy was checked by the recovery studies conducted at three different levels with the known concentrations and the average percentage recovery was 101.99% for AN and 103.84% for AA. The precision was analyzed by interday and intraday precision and was 1.09 and 1.00% RSD for AN and 1.22 and 6.05% RSD for AA. The analysis of statistical data substantiates that this HPTLC method can be used for the simultaneous estimation of AN and AA in biological samples. Copyright © 2015 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

We isolated a 2-alkenal reductase (AaAER) gene from Artemisia annua L. cDNA library through homologous cloning strategy. Enzymatic properties of AaAER, including substrate selectivity, enzyme kinetics, and key factors affecting enzyme activity, were characterized in vitro. We found that AaAER mainly functions in catalyzing the reduction of adjacent straight chain 'C = C' double bond in the carbonyl group. This suggests the possibility of its participation in downstream events of the HPL (hydroperoxide lyase) pathway and contribution to cell detoxification in plants. Although, AaAER shares some substrates with AaDbr 1 and AaDbr 2, which are involved in artemisinin biosynthesis, AaAER was unable to reduce the adjacent double bond of carbonyl group in artemisinic aldehyde. We also analyzed the possible causes for highly homologous reductases with different types of substrates and non-homologous enzymes sharing the same substrate. The identification of a novel double bond reductase in A. annua provides an opportunity to explore the mechanism underlying the detoxification of aldehyde ketone molecules, which are generated by HPL downstream pathway and the redox process in the metabolic pathway of artemisinin synthesis. [ABSTRACT FROM AUTHOR]

Artemisinic aldehyde Δ11 (13) reductase ( DBR2) is the checkpoint enzyme catalyzing artemisinic aldehyde to form dihydroartemisinic aldehyde directly involved in artemisinin biosynthetic pathway. In the present study, DBR2 was employed to engineer the biosynthetic pathway of artemisinin in transgenic plants of Artemisia annua L. Seven independent transgenic plants of A. annua with DBR2 overexpression driven by the cauliflower mosaic virus 35S promoter were obtained by Agrobacterium-mediated genetic transformation and confirmed by genomic PCR. The results of real-time q PCR analysis showed that the expression levels of DBR2 gene in all the seven transgenic lines were significantly higher than in nontransgenic control. The high-performance liquid chromatography analysis of artemisinin and its relative metabolites demonstrated that the contents of artemisinin and its direct precursor dihydroartemisinic acid were remarkably increased in the transgenic plants of A. annua with DBR2 overexpression. Interestingly, it was also found that the contents of arteannuin B and its direct precursor artemisinic acid in the branch pathway competing against artemisinin biosynthesis were also improved in DBR2-overexpressed A. annua plants. The transgenic results in the present study indicated that DBR2 is a useful structural gene in engineering the artemisinin biosynthetic pathway to develop genetically modified A. annua with the higher yield of artemisinin. [ABSTRACT FROM AUTHOR]

Amorpha-4,11-diene synthase (ADS) and Cyt P450 monooxygenase (CYP71AV1) in Artemisia annua L. are two key enzymes involved in the biosynthesis of artemisinin. The promoters of ADS and CYP71AV1 contain E-box elements, which are putative binding sites for basic helix–loop–helix (bHLH) transcription factors. This study successfully isolated a bHLH transcription factor gene from A. annua, designated as AabHLH1, from a cDNA library of the glandular secretory trichomes (GSTs) in which artemisinin is synthesized and sequestered. AabHLH1 encodes a protein of 650 amino acids containing one putative bHLH domain. AabHLH1 and ADS genes were strongly induced by ABA and the fungal elicitor, chitosan. The transient expression analysis of the AabHLH1–green fluorescent protein (GFP) reporter gene revealed that AabHLH1 was targeted to nuclei. Biochemical analysis demonstrated that the AabHLH1 protein was capable of binding to the E-box cis-elements, present in both ADS and CYP71AV1 promoters, and possessed transactivation activity in yeast. In addition, transient co-transformation of AabHLH1 and CYP71AV1Pro::GUS in A. annua leaves showed a significant activation of the expression of the GUS (β-glucuronidase) gene in transformed A. annua, but mutation of the E-boxes resulted in abolition of activation, suggesting that the E-box is important for the CYP71AV1 promoter activity. Furthermore, transient expression of AabHLH1 in A. annua leaves increased transcript levels of the genes involved in artemisinin biosynthesis, such as ADS, CYP71AV1 and HMGR. These results suggest that AabHLH1 can positively regulate the biosynthesis of artemisinin. [ABSTRACT FROM PUBLISHER]

Artemisinin is a compound extracted fromArtemisia annuaL. with a remarkable curative effect against malaria. It can be extracted using ultrasound-assisted extraction (UAE) and then detected via HPLC. In this study, response surface methodology (RSM) was used to optimize UAE conditions for obtaining the maximum yield of artemisinin. Three independent variables (ratio of solvent to material, extraction temperature, and ultrasonic power) were evaluated using the Box-Behnken experimental design, with the yield of artemisinin as a response variable. Experimental data were highly fitted to a mathematical-regression model using multiple linear regression (MLR). Based on response surface plots, the three independent variables exhibited interactive effects on the yield of artemisinin. The optimal extraction conditions were as follows: 42.71 mL/g ratio of solvent to material, 41.86°C extraction temperature and 120 W ultrasonic power. The predicted yield of artemisinin by model was 0.7848%, whereas the actual yield in the extracts was 0.7826% ± 0.0790% in adjusted optimal conditions, with a relative error of 0.28%. The results undoubtedly demonstrated that RSM could be used to explore the optimum conditions of artemisinin extraction. [ABSTRACT FROM AUTHOR]