Potential of herbaceous plant species for copper (Cu) accumulation.

The removal of copper (Cu) in soils by green technology is less treated with urgency, as it is a plant micronutrient. We examined the efficiency of Cu shoot accumulation by herbaceous plants in Cu-contaminated and non-contaminated soils in Trhové Dusniky and Podles, respectively, in the Czech Republic. The total soil Cu content of 81 mg kg-1 in Trhové Dusniky indicated a slight contamination level compared to 50 mg kg-1, the permissible value by WHO, and < 35 in Podlesí, representing a clean environment. The Cu content was above the permissible value in plants (10 mg kg-1 by WHO) in herbaceous speciesat the control site without trees: Stachys palustris L. (10.8 mg kg-1), Cirsium arvense L. (11.3 mg kg-1), Achillea millefolium L. (12.1 mg kg-1), Anthemis arvense L. (13.2 mg kg-1), and Calamagrostis epigejos L. (13.7 mg kg-1). In addition, Hypericum maculatum Crantz (10.6 mg kg-1), Campanula patula L. (11.3 mg kg-1), C. arvense (15 mg kg-1), and the highest accumulation in shoot of Equisetum arvense L. (37.1 mg kg-1), all under the canopy of trees at the uncontaminated site, were above the WHO value. Leucanthemum Vulgare (Lam.) and Plantago lanceolata L. recorded 11.2 mg kg-1 and 11.5 mg kg-1, respectively, in the soil of the Cu-contaminated site. These herbaceous species can support the phyto-management of Cu-contaminated soils, especially E. arvense. Critical attention is well-required in the medicinal application of herbaceous plants in treating human ailments due to their Cu accumulation potentials above the threshold. Spontaneous surveys and analysis of Cu speciation in herbaceous species can reveal suitable plants to decontaminate soils and provide caution on consumable products, especially bioactive compounds.

Research progress on the development of pennycress (Thlaspi arvense L.) as a new seed oil crop: a review.

Compared with other crops, pennycress (Thlaspi arvense L.) is a niche emerging oil crop. In recent years, research on pennycress has been increasingly reflected in various directions. Pennycress belongs to the Brassicaceae family and was introduced from Eurasia to North America. It has been found worldwide as a cultivated plant and weed. In this paper, we review the advantages of pennycress as a supplementary model plant of Arabidopsis thaliana, oil and protein extraction technology, seed composition analysis based on metabolomics, germplasm resource development, growth, and ecological impact research, abiotic stress, fatty acid extraction optimization strategy, and other aspects of studies over recent years. The main research directions proposed for the future are as follows: (1) assemble the genome of pennycress to complete its entire genome data, (2) optimize the extraction process of pennycress as biodiesel, (3) analyze the molecular mechanism of the fatty acid synthesis pathway in pennycress, and (4) the functions of key genes corresponding to various adversity conditions of pennycress.

Phytotoxicity of Extracts of Argemone mexicana and Crotalaria longirostrata on Tomato Seedling Physiology.

Phytotoxicity caused by secondary metabolites of botanical extracts is a drawback in agriculture. The objective of this study was to evaluate the phytotoxic effects of methanolic extracts of Crotalaria longirostrata and Argemone mexicana on the germination and physiological variables of tomato seedlings. The results indicated that high doses of both extracts (Clong500 and Amex500) inhibited tomato seed germination, while their mixture (Cl50 + Am50) promoted germination by 100%. At 30 days after transplanting (dat), the plant height increased by 15.4% with a high dose of C. longirostrata (Clong500) compared to the control. At 30 dat, the vigor index displayed a notable increase with Cl50 + Am50, reaching 29.5%. The root length increased with the mean dose of A. mexicana (Amex95) at 10, 20, and 30 dat (59.7%, 15.1%, and 22.4%, respectively). The chlorophyll content increased with Amex95 by 66.1% in 10 dat, 22.6% at 20 dat, and 19.6% at 30 dat. On the other hand, Amex95 had a higher nitrogen content throughout the trial. Amex95 produced the greatest increase in root dry weight by 731.5% and 209.4% at 10 and 20 dat. The foliage dry weight increased by 85.7% at 10 dat with Amex95 and up to 209.7% with Amex50 at 30 dat. The present investigation reveals the ability of the extracts to stimulate tomato growth at low and medium doses, though at high doses they exhibit allelopathic effects.

Is pea our hidden allergen? An American pediatric case series.

Plant-based diets, consisting of legumes, are becoming increasingly a diet trend, thus a focus for food manufacturers as a source of protein. Allergy to legumes, specifically to green pea and dun pea, have been emerging. There is currently no data on pea allergy in the United States. As such, and with the progressive increase in pea/pea proteins inclusion into foods, we present in this case series children with allergic reactions to foods containing green peas or pea-protein.

Phytochemical Investigation of Equisetum arvense and Evaluation of Their Anti-Inflammatory Potential in TNFα/INFγ-Stimulated Keratinocytes.

Equisetum arvense L. (Equisetaceae), widely known as 'horsetail', is a perennial plant found extensively across Asia. Extracts of E. arvense have been used in traditional medicine, particularly for the treatment of inflammatory disorders. This study aimed to determine the phytochemical compounds in E. arvense ethanolic extract and their anti-inflammatory properties. Subsequently, we isolated and identified nine secondary metabolites, including kaempferol 3,7-di-O-ß-D-glucopyranoside (1), icariside B2 (2), (Z)-3-hexenyl ß-D-glucopyranoside (3), luteolin 5-O-ß-D-glucopyranoside (4), 4-O-ß-D-glucopyranosyl caffeic acid (5), clemastanin B (6), 4-O-caffeoylshikimic acid (7), (7S,8S)-threo-7,9,9'-trihydroxy-3,3'-dimethoxy-8-O-4'-neolignan-4-O-ß-D-glucopyranoside (8), and 3-O-caffeoylshikimic acid (9). The chemical structures of the isolated compounds (1-9) were elucidated using HR-ESI-MS data, NMR spectra, and ECD data. Next, the anti-inflammatory effects of the isolates were evaluated in tumor necrosis factor (TNF)α/interferon (IFN)γ-induced HaCaT, a human keratinocyte cell line. Among the isolates, compound 3 showed the highest inhibitory effect on the expression of pro-inflammatory chemokines, followed by compounds 6 and 8. Correspondingly, the preceding isolates inhibited TNFα/IFNγ-induced activation of pro-inflammatory transcription factors, signal transducer and activator of transcription 1, and nuclear factor-κB. Collectively, E. arvense could be employed for the development of prophylactic or therapeutic agents for improving dermatitis.

Equisetum arvense L aqueous extract: a novel chemotherapeutic supplement for treatment of human colon carcinoma.

Introduction: One of the plants that has long been considered by humans is Equisetum arvense L. Equisetum arvense L is now recommended for external use to heal wounds and for internal use to relieve urinary tract and prostate disorders. In the current study, the antioxidant, cytotoxicity, and anti-human lung cancer properties of Equisetum arvense were investigated in in vitro conditions. Material and methods: Total phenolic content, total flavonoid content, radical scavenging activity, and ferrous ion chelating were assessed to evaluate the antioxidant activity. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was chosen to investigate anticancer activity of the plant extract. Results: The plant extract scavenged 2,2-diphenyl-1-picrylhydrazyl (DPPH) as a free radical with an IC50 of 12.3 ±0.7 µg/ml better than positive controls. The plant was also rich in phenolic compounds with an amount of 396.2 ±3.2 mg GAE/g for total phenolic content. In the MTT assay, human colorectal carcinoma (HCT-8 [HRT-18], Ramos.2G6.4C10, HT-29, and HCT 116) and normal cell lines (HUVEC) were used to study the cytotoxicity and anticancer potential of Equisetum arvense L against human colorectal cancer. Conclusions: The cell viability of Equisetum arvense L was very low against human colorectal carcinoma cell lines without any cytotoxicity towards the normal (HUVEC) cell line. The best anti-human colorectal carcinoma properties of Equisetum arvense L against the above cell lines were observed in the case of the HT 29 cell line.

Metabolic and transcriptomic study of pennycress natural variation identifies targets for oil improvement.

Pennycress (Thlaspi arvense L.), a member of the Brassicaceae family, produces seed oil high in erucic acid, suitable for biodiesel and aviation fuel. Although pennycress, a winter annual, could be grown as a dedicated bioenergy crop, an increase in its seed oil content is required to improve its economic competitiveness. The success of crop improvement relies upon finding the right combination of biomarkers and targets, and the best genetic engineering and/or breeding strategies. In this work, we combined biomass composition with metabolomic and transcriptomic studies of developing embryos from 22 pennycress natural variants to identify targets for oil improvement. The selected accession collection presented diverse levels of fatty acids at maturity ranging from 29% to 41%. Pearson correlation analyses, weighted gene co-expression network analysis and biomarker identifications were used as complementary approaches to detect associations between metabolite level or gene expression and oil content at maturity. The results indicated that improving seed oil content can lead to a concomitant increase in the proportion of erucic acid without affecting the weight of embryos. Processes, such as carbon partitioning towards the chloroplast, lipid metabolism, photosynthesis, and a tight control of nitrogen availability, were found to be key for oil improvement in pennycress. Besides identifying specific targets, our results also provide guidance regarding the best timing for their modification, early or middle maturation. Thus, this work lays out promising strategies, specific for pennycress, to accelerate the successful development of lines with increased seed oil content for biofuel applications.

3D-Printed PLA Molds for Natural Composites: Mechanical Properties of Green Wax-Based Composites.

The first part of this paper is dedicated to obtaining 3D-printed molds using poly lactic acid (PLA) incorporating specific patterns, which have the potential to serve as the foundation for sound-absorbing panels for various industries and aviation. The molding production process was utilized to create all-natural environmentally friendly composites. These composites mainly comprise paper, beeswax, and fir resin, including automotive function as the matrices and binders. In addition, fillers, such as fir needles, rice flour, and Equisetum arvense (horsetail) powder, were added in varying amounts to achieve the desired properties. The mechanical properties of the resulting green composites, including impact and compressive strength, as well as maximum bending force value, were evaluated. The morphology and internal structure of the fractured samples were analyzed using scanning electron microscopy (SEM) and an optical microscopy. The highest impact strength was measured for the composites with beeswax, fir needles, recyclable paper, and beeswax fir resin and recyclable paper, 19.42 and 19.32 kJ/m2, respectively, while the highest compressive strength was 4 MPa for the beeswax and horsetail-based green composite. Natural-material-based composites exhibited 60% higher mechanical performance compared to similar commercial products used in the automotive industry.

Invasive Cirsium arvense displays different resource-use strategies along local habitat heterogeneity in the trans-Himalayan region of Ladakh.

Climate change and anthropogenic pressures have resulted in a significant shift in the invasion susceptibility and frequency of non-native species in mountain ecosystems. Cirsium arvense (L.) Scop. (Family: Asteraceae) is an invasive species that spreads quickly in mountains, especially in the trans-Himalayan region of Ladakh. The current study used a trait-based approach to evaluate the impact of local habitat heterogeneity (soil physico-chemical properties) on C. arvense. Thirteen plant functional traits (root, shoot, leaf, and reproductive traits) of C. arvense were studied in three different habitat types (agricultural, marshy, and roadside). Functional trait variability in C. arvense was higher between, than within habitats (between different populations). All the functional traits interacted with habitat change, except for leaf count and seed mass. Soil properties strongly affect C. arvense's resource-use strategies across habitats. The plant adapted to a resource-poor environment (roadside habitat) by conserving resources and to a resource-rich environment (agricultural and marshy land habitat) by acquiring them. The ability of C. arvense to use resources differently reflects its persistence in introduced habitats. In summary, our study shows that C. arvense invades different habitats in introduced regions through trait adaptations and resource-use strategies in the trans-Himalayan region.

Effects of Phytotoxic Nonenolides, Stagonolide A and Herbarumin I, on Physiological and Biochemical Processes in Leaves and Roots of Sensitive Plants.

Phytotoxic macrolides attract attention as prototypes of new herbicides. However, their mechanisms of action (MOA) on plants have not yet been elucidated. This study addresses the effects of two ten-membered lactones, stagonolide A (STA) and herbarumin I (HBI) produced by the fungus Stagonospora cirsii, on Cirsium arvense, Arabidopsis thaliana and Allium cepa. Bioassay of STA and HBI on punctured leaf discs of C. arvense and A. thaliana was conducted at a concentration of 2 mg/mL to evaluate phenotypic responses, the content of pigments, electrolyte leakage from leaf discs, the level of reactive oxygen species, Hill reaction rate, and the relative rise in chlorophyll a fluorescence. The toxin treatments resulted in necrotic and bleached leaf lesions in the dark and in the light, respectively. In the light, HBI treatment caused the drop of carotenoids content in leaves on both plants. The electrolyte leakage caused by HBI was light-dependent, in contrast with that caused by STA. Both compounds induced light-independent peroxide generation in leaf cells but did not affect photosynthesis 6 h after treatment. STA (10 µg/mL) caused strong disorders in root cells of A. thaliana leading to the complete dissipation of the mitochondrial membrane potential one hour post treatment, as well as DNA fragmentation and disappearance of acidic vesicles in the division zone after 8 h; the effects of HBI (50 µg/mL) were much milder. Furthermore, STA was found to inhibit mitosis but did not affect the cytoskeleton in cells of root tips of A. cepa and C. arvense, respectively. Finally, STA was supposed to inhibit the intracellular vesicular traffic from the endoplasmic reticulum to the Golgi apparatus, thus interfering with mitosis. HBI is likely to have another main MOA, probably inhibiting the biosynthesis of carotenoids.

Novel Approach to the Treatment of Neuropathic Pain Using a Combination with Palmitoylethanolamide and Equisetum arvense L. in an In Vitro Study.

Neuropathic pain is a typical patient disorder resulting from damage and dysfunction of the peripheral neuraxis. Injury to peripheral nerves in the upper extremities can result in a lifelong reduction in quality of life and a devastating loss of sensory and motor function. Since some standard pharmaceutical therapies can cause dependence or intolerance, nonpharmacological treatments have gained great interest in recent years. In this context, the beneficial effects of a new combination of palmitoylethanolamide and Equisetum arvense L. are evaluated in the present study. The bioavailability of the combination was initially analyzed in a 3D intestinal barrier simulating oral intake to analyze its absorption/biodistribution and exclude cytotoxicity. In a further step, a 3D nerve tissue model was performed to study the biological effects of the combination during the key mechanisms leading to peripheral neuropathy. Our results demonstrate that the combination successfully crossed the intestinal barrier and reached the target site, modulating the nerve recovery mechanism after Schwann cell injury and offering the initial response of relieving pain. This work supported the efficacy of palmitoylethanolamide and Equisetum arvense L. in reducing neuropathy and modifying the major pain mechanisms, outlining a possible alternative nutraceutical approach.

Morphologically and genetically diverse forage pea (Pisum sativum var. arvense L.) genotypes under single and combined salt and drought stresses.

Salinity and drought stresses limit agricultural productivity of many crops including forage pea which is an important forage legume. Due to increasing importance of legumes in forage production, there is a clear need to investigate the underlying affects of salinity and drought stresses on forage pea. This study was designed to understand how single or combined salinity and drought stresses impact on physio-biochemical and molecular status of morphologically and genetically diverse forage pea genotypes. Firstly, yield-related parameters were determined under three-year field experiment. The results revealed that the agro-morphological features of the genotypes are significantly different. Afterwards, the sensitivities of the 48 forage pea genotypes were determined against single and combined salinity and drought stresses by performing growth parameters, biochemical status, antioxidative enzymes, and endogenous hormones. Also, the salt and drought-related gene expressions were evaluated under normal and stressed conditions. The results collectively showed that the genotypes of O14, and T8 were more tolerant against combined stress compared to others, via activating antioxidative enzymes (CAT, GR, and SOD), endogenous hormones (IAA, ABA, and JA), stress-related genes (DREB3, DREB5, bZIP11, bZIP37, MYB48, ERD, RD22) and leaf senescence genes (SAG102, SAG102). These genotypes could be used to develop pea plants that tolerate salinity or drought stress conditions. To the best of our knowledge, the present study is the first detailed study in pea against combined salt and drought stresses.

Functional Properties and Acceptability of Potentially Medicinal Tea Infusions Based on Equisetum arvense, Desmodium molliculum, and Mentha piperita.

Natural herbal teas are one of the three most consumed beverages in the world, and despite their frequent use in the cosmetic, food, and pharmaceutical industries, there is still much to about them. This study aimed to determine the functional properties of tea infusions made from dried Equisetum arvense (EA), Desmodium molliculum (DM), and Mentha piperita (M) grown in the Peruvian Andes. Next, using a simplex design with unrestricted centroid amplified centroid, 12 combinations were obtained for the combination of dried leaves with EA: 0∼100%, DM: 0∼100%, and M: 0∼100% optimal combination of EA: 6.59%, DM: 84.62%, and M: 8.79% maximizes functional components for total polyphenols (2,831.18 mg EAG/100 g), flavonoids (37.73 mg CAT/g), and antioxidant capacity (145.99 µmol Trolox/g). It can be confirmed that dried mixtures of these plants made into tea are a significant source of bioactive molecules, have a tolerable flavor, and can be used for therapeutic purposes when consumed.

A lipidomics platform to analyze the fatty acid compositions of non-polar and polar lipid molecular species from plant tissues: Examples from developing seeds and seedlings of pennycress (Thlaspi arvense).

The lipidome comprises the total content of molecular species of each lipid class, and is measured using the analytical techniques of lipidomics. Many liquid chromatography-mass spectrometry (LC-MS) methods have previously been described to characterize the lipidome. However, many lipidomic approaches may not fully uncover the subtleties of lipid molecular species, such as the full fatty acid (FA) composition of certain lipid classes. Here, we describe a stepwise targeted lipidomics approach to characterize the polar and non-polar lipid classes using complementary LC-MS methods. Our "polar" method measures 260 molecular species across 12 polar lipid classes, and is performed using hydrophilic interaction chromatography (HILIC) on a NH2 column to separate lipid classes by their headgroup. Our "non-polar" method measures 254 molecular species across three non-polar lipid classes, separating molecular species on their FA characteristics by reverse phase (RP) chromatography on a C30 column. Five different extraction methods were compared, with an MTBE-based extraction chosen for the final lipidomics workflow. A state-of-the-art strategy to determine and relatively quantify the FA composition of triacylglycerols is also described. This lipidomics workflow was applied to developing, mature, and germinated pennycress seeds/seedlings and found unexpected changes among several lipid molecular species. During development, diacylglycerols predominantly contained long chain length FAs, which contrasted with the very long chain FAs of triacylglycerols in mature seeds. Potential metabolic explanations are discussed. The lack of very long chain fatty acids in diacylglycerols of germinating seeds may indicate very long chain FAs, such as erucic acid, are preferentially channeled into beta-oxidation for energy production.

A Multi-Pronged Technique for Identifying Equisetum palustre and Equisetum arvense-Combining HPTLC, HPLC-ESI-MS/MS and Optimized DNA Barcoding Techniques.

The most prominent horsetail species, Equisetum arvense, has an array of different medicinal properties, thus the proper authentication and differentiation of the plant from the more toxic Equisetum palustre is important. This study sought to identify different samples of E. arvense and E. palustre using three analytical methods. The first method involved the use of HPTLC analysis, as proposed by the European Pharmacopoeia. The second, HPLC-ESI-MS/MS, is capable of both identification and quantification and was used to determine the Equisetum alkaloid content in each sample. A third method was DNA barcoding, which identifies the samples based on their genetic make-up. Both HPTLC and HPLC-ESI-MS/MS proved to be suitable methods of identification, with HPLC-ESI-MS/MS proving the more sophisticated method for the quantification of alkaloids in the Equisetum samples and for determining the adulteration of E. arvense. For DNA barcoding, optimal primer pairs were elucidated to allow for the combined use of the rbcL and ITS markers to accurately identify each species. As new DNA marker sequences were added to GenBank, the reference library has been enriched for future work with these horsetail species.

Effective Mechanisms for Improving Seed Oil Production in Pennycress (Thlaspi arvense L.) Highlighted by Integration of Comparative Metabolomics and Transcriptomics.

Pennycress is a potentially lucrative biofuel crop due to its high content of long-chain unsaturated fatty acids, and because it uses non-conventional pathways to achieve efficient oil production. However, metabolic engineering is required to improve pennycress oilseed content and make it an economically viable source of aviation fuel. Research is warranted to determine if further upregulation of these non-conventional pathways could improve oil production within the species even more, which would indicate these processes serve as promising metabolic engineering targets and could provide the improvement necessary for economic feasibility of this crop. To test this hypothesis, we performed a comparative biomass, metabolomic, and transcriptomic analyses between a high oil accession (HO) and low oil accession (LO) of pennycress to assess potential factors required to optimize oil content. An evident reduction in glycolysis intermediates, improved oxidative pentose phosphate pathway activity, malate accumulation in the tricarboxylic acid cycle, and an anaplerotic pathway upregulation were noted in the HO genotype. Additionally, higher levels of threonine aldolase transcripts imply a pyruvate bypass mechanism for acetyl-CoA production. Nucleotide sugar and ascorbate accumulation also were evident in HO, suggesting differential fate of associated carbon between the two genotypes. An altered transcriptome related to lipid droplet (LD) biosynthesis and stability suggests a contribution to a more tightly-packed LD arrangement in HO cotyledons. In addition to the importance of central carbon metabolism augmentation, alternative routes of carbon entry into fatty acid synthesis and modification, as well as transcriptionally modified changes in LD regulation, are key aspects of metabolism and storage associated with economically favorable phenotypes of the species.

Soybean Cyst Nematode Population Development and its Effect on Pennycress in a Greenhouse Study.

Midwest crop production is dominated by two summer annual crops grown in rotation, viz., corn (Zea mays L.) and soybean (Glycine max L.). Winter oilseed crops, such as pennycress (Thlaspi arvense L.), can provide ecosystem and economic benefits when added to the corn-soybean rotation. However, adding a new crop adds risks, such as increased pest pressure. The objectives of this study were to (i) evaluate population development of three soybean cyst nematode (SCN; Heterodera glycines) biotypes on three pennycress genotypes and susceptible soybean and (ii) determine whether SCN inoculation level influenced plant biomass. SCN population density and biomass were determined after 60 d in the greenhouse. At the inoculation level of 2,000 eggs/100 cm3 soil, the average egg density for the three pennycress genotypes was 1,959 eggs/100 cm3 soil, lower than that for the susceptible soybean 'Sturdy' (9,601 eggs/100 cm3 soil). At the inoculation level of 20,000 eggs/100 cm3 soil, the average egg density for the three pennycress genotypes was 6,668 eggs/100 cm3 soil, lower than that for 'Sturdy' (40,740 eggs/100 cm3 soil). The inoculation level did not affect plant biomass. Pennycress is an alternative host to SCN under greenhouse conditions but is a less suitable host than soybean.

Aqueous extract from Equisetum arvense stimulates the secretion of Tamm-Horsfall protein in human urine after oral intake.

BACKGROUND: Within European traditional phytotherapy, extracts from different herbal plants are used for prevention and therapy of uncomplicated urinary tract infections and for flushing out of kidney grits. Besides increased urine flow by slight diuretic effects, also stimulation of Tamm-Horsfall protein (syn. THP, uromodulin) in the distal part of the kidney could explain reduced kidney gravel and anti-virulent activity against uropathogenic E. coli. PURPOSES: Evaluation of THP-inducing activity of extracts from Equisetum arvense, Levisticum officinalis, Ilex paraguariensis, Juniperus communis, Urtica dioica, and Taraxacum officinale by quantification of THP in urine samples after oral application to humans. STUDY DESIGN: 7 days p.o. application of the test intervention to healthy volunteers (n = 10 per intervention group) and analysis of urine samples at day 1 (untreated control values), and days 3, 6 and 8 on THP content by validated ELISA. Antiadhesive activity of urine samples was monitored by flow cytometry using UPEC strain NU14 against human T24 bladder cells. RESULTS: An aqueous extract from E. arvense, fully characterized by a specific LC-MS method, induced THP concentration in urine samples significantly during a 7-day p.o. application up to 300%, related to the untreated controls. Ex vivo investigation of the individual and pooled urine samples with elevated THP concentrations showed good correlation to antiadhesive effects against UPEC NU14 to T24 cells. Urine samples of the Equisetum treated volunteers had no effect on the proliferation and on biofilm formation of UPEC NU14. Silica excretion in the urine samples had no correlation to the respective THP levels. Monitoring of electrolyte content in the urine samples indicat ed diuretic effects of the intervention with Equisetum extract. Detailed phytochemical analysis of the Equisetum extract by LC-MS and LC-UV revealed an analytical protocol, which identified > 80 compounds from the extract by MS evaluations and 18 compounds by UV detection. This protocol will provide a valuable tool for future quality control of Equisetum extract. CONCLUSION: Aqueous extract from E. arvense significantly stimulates THP secretion in urine samples after 7 days of oral intake and inhibits the interplay between UPEC and bladder host cells. This could explain the therapeutic use of this herbal material for urinary tract infections and kidney gravel. Detailed phytochemical analysis of the Equisetum extract by LC-MS and LC-UV revealed an analytical protocol, which identified > 82% of all eluted compounds. This protocol will provide a valuable tool for future quality control of Equisetum extract.

Investigation of the Genotoxic, Antigenotoxic and Antioxidant Profile of Different Extracts from Equisetum arvense L.

The present study investigated the cyto-genotoxic and antigenotoxic effects of four different extracts of Equisetum arvense L. (common name: field horsetail) on human lymphocytes. Specifically, Soxhlet's prepared extracts from E. arvense L., using different solvents (S1: methanol (MeOH)-, S2: ethanol (EtOH)-, S3: water-, and S4: ethanol/water (EtOH-W)-) were analyzed for (a) their total phenolic and flavonoid content (TPC and TFC, respectively), (b) their antioxidant activity (AA), via the DPPH, FRAP and ABTS assays, and (c) their cyto-genotoxic and/or protective efficiency against the mutagenic agent mitomycin C, via the Cytokinesis Block MicroNucleus assay. All extracts showed increased TPC, TFC, and AA values in almost all cases. S1, S3 and S4 demonstrated no cytotoxic potential, whereas S2 was cytotoxic only at the highest concentrations. Genotoxicity was not observed in the tested extracts. The highest antigenotoxic activity was observed for EtOH-W (S4) extract, which was found to be rich in flavonoids, flavonoid-O-glycosides, phytosterols, phenolic and fatty acids as well as in minerals and mainly in K, Ca, Mg, Si and P, as assessed by using various mass spectrometry techniques. Those findings confirm that E. arvense L. extracts could be valuable candidates for medicinal applications and pharmaceutical products, thus alleviating the effects of more conventional drugs.

Structure, mechanism, and phylogeny of LysM-chitinase conjugates specifically found in fern plants.

A unique GH18 chitinase containing two N-terminal lysin motifs (PrLysM1 and PrLysM2) was first found in fern, Pteris ryukyuensis (Onaga and Taira, Glycobiology, 18, 414-423, 2008). This type of LysM-chitinase conjugates is not usually found in plants but in fungi. Here, we produced a similar GH18 chitinase with one N-terminal LysM module (EaLysM) from the fern, Equisetum arvense (EaChiA, Inamine et al., Biosci. Biotechnol. Biochem., 79, 1296-1304, 2015), using an Escherichia coli expression system and characterized for its structure and mechanism of action. The crystal structure of EaLysM exhibited an almost identical fold (ßααß) to that of PrLysM2. From isothermal titration calorimetry and nuclear magnetic resonance, the binding mode and affinities of EaLysM for chitooligosaccharides (GlcNAc)n (3, 4, 5, and 6) were found to be comparable to those of PrLysM2. The LysM module in EaChiA is likely to bind (GlcNAc)n almost independently through CH-π stacking of a Tyr residue with the pyranose ring. The (GlcNAc)n-binding mode of LysMs in the LysM-chitinase conjugates from fern plants appears to differ from that of plant LysMs acting in chitin- or Nod-signal perception, in which multiple LysMs cooperatively act on (GlcNAc)n. Phylogenetic analysis suggested that LysM-GH18 conjugates of fern plants formed a monophyletic group and had been separated earlier than forming the clade of fungal chitinases with LysMs.