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Fucoidans, a diverse group of sulphated polysaccharides from brown algae, are biopolymers with many documented activities in vitro. While several factors are known to influence these activities, the degree of sulphation of the polymer is often regarded as a key parameter. The degree of sulphation on a biopolymer may be seasonally dependent, which, in turn, may affect the bioactivities of fucoidans. In this study, the effect of season and geographical location on the fucoidan sulphur content (as a proxy for degree of sulphation), was investigated. Crude fucoidan extracts were obtained using microwave-assisted extraction on eight different brown algae species, which were then analysed using elemental analysis. A significant seasonal dependence on the sulphur content in fucoidan extracts from Fucus serratus was observed. The difference in sulphur content between the investigated Fucus species is likely due to their different reproduction strategies. Of the eight species used in this study, Ascophyllum nodosum and Fucus evanescens contained the most sulphated fucoidans, while the species Dictyosiphon foeniculaceus contained lightly sulphated fucoidans. This suggests that species which thrive in saline waters may contain more sulphated fucoidans as a means of mitigating osmotic stress. No significant effects on the sulphur content in fucoidans from Fucus vesiculosus was observed between the three investigated countries. From these findings, it is apparent that seasonality is an important parameter, which must be considered for large scale production of highly sulphated fucoidans.

The need for a substitute for allograft and autograft is rising as bone graft surgeries exceed available supplies. We investigated the efficacy of the low-molecular weight marine bioactive compound fucoidan (FUC) on bone regeneration and implant fixation in seven female sheep, as FUC has shown great promise as a bone substitute. Titanium implants were inserted bilaterally in the distal femurs to test three hydroxyapatite/fucoidan (HA/FUC) groups and compared to allograft. The HA was coated with either 500 or 1500 μg of FUC, obtained by microwave-assisted chemical extraction, or 500 μg of FUC obtained by an enzyme-assisted extraction method. The concentric 2-mm gap around the implant was filled with either one of the HA/FUCs or allograft from the donor sheep. After 12 weeks, implant–bone blocks were harvested and divided into three parts for mechanical push-out testing, immunohistochemistry, and micro-CT and histomorphometry. Pronounced bone formations were observed by micro-CT and histomorphometry in all groups, but higher bone volume fractions were seen in the allograft group compared to the three HA/FUC groups. The trabecular thickness, trabecular separation, and architectural anisotropy were all significantly higher in the allograft group compared to the three HA/FUC groups. In conclusion, adequate bone formation was observed in all groups, although the bone formation was significantly greater in the allograft group. Also, no significant differences existed in the shear mechanical properties between groups, suggesting that the combination of HA and FUC can achieve a similar fixation strength to allograft in this model.

In drug development, estimating fraction absorbed (F a) in man for permeability-limited compounds is important but challenging. To model F a of such compounds from apparent permeabilities (P app) across filter-grown Caco-2 cell monolayers, it is central to elucidate the intestinal permeation mechanism(s) of the compound. The present study aims to refine a computational permeability model to investigate the relative contribution of paracellular and transcellular routes to the P app across Caco-2 monolayers of the permeability-limited compound acamprosate having a bioavailability of ∼11%. The P app values of acamprosate and of several paracellular marker molecules were measured. These P app values were used to refine system-specific parameters of the Caco-2 monolayers, that is, paracellular pore radius, pore capacity, and potential drop. The refined parameters were subsequently used as an input in modeling the permeability (P modeled) of the tested compounds using mathematical models collected from two published permeability models. The experimental data show that acamprosate P app across Caco-2 monolayers is low and similar in both transport directions. The obtained acamprosate P app, 1.56 ± 0.28 × 10 -7 cm·s -1, is similar to the P app of molecular markers for paracellular permeability, namely, mannitol (2.72 ± 0.24 × 10 -7 cm·s -1), lucifer yellow (1.80 ± 0.35 × 10 -7 cm·s -1), and fluorescein (2.10 ± 0.28 × 10 -7 cm·s -1), and lower than that of atenolol (7.32 ± 0.60 × 10 -7 cm·s -1 mean ± SEM, n = 3-6), while the end-point amount of acamprosate internalized by the cell monolayer, Q monolayer, was lower than that of mannitol. Acamprosate did not influence the barrier function of the monolayers since it altered neither the P app of the three paracellular markers nor the transepithelial electrical resistance (TEER) of the cell monolayer. The P modeled for all the paracellular markers and acamprosate was dominated by the P para component and matched the experimentally obtained P app. Furthermore, acamprosate did not inhibit the uptake of probe substrates for solute carriers PEPT1, TAUT, PAT1, EAAT1, B 0,+AT/rBAT, OATP2B1, and ASBT expressed in Caco-2 cells. Thus, the P modeled estimated well P para, and the paracellular route appears to be the predominant mechanism for acamprosate P app across Caco-2 monolayers, while the alternative transcellular routes, mediated by passive diffusion or carriers, are suggested to only play insignificant roles.

Vegetables rich in bitter-tasting phytochemicals may exert enhanced beneficial effects against key factors associated with type two diabetes (T2D). This study investigates whether selected cultivars of bitter and strong-tasting (BST) Brassica and root vegetables exert greater health benefits on T2D patients compared to equivalent modern mild and sweet tasting (MST) vegetables. A 12-week randomized, controlled, parallel intervention study involved 92 T2D patients, who were allocated three different diets: (1) 500 g daily of bitter and strong-tasting (BST) vegetables, (2) 500 g daily of mild and sweet-tasting (MST) vegetables, (3) 120 g daily MST normal diet (control). Both vegetable diets contained root vegetables and cabbages selected based on sensory differences and content of phytochemicals. Prior to and after the study, all participants underwent an oral glucose tolerance test (OGTT), 24 h blood pressure measurements, DEXA scans, and fasted blood samples. Both diets high in vegetables significantly reduced the participants’ BMI, total body fat mass, and HbA1c levels compared to control, but in the BST group, significant differences were also found regarding incremental area under the curve glucose 240 min (OGTT) and fasting glucose levels. A high daily intake of root vegetables and cabbages showed significant health improvements in both vegetable groups. BST vegetables had the greatest impact on insulin sensitivity, body fat mass, and blood pressure compared to control, moreover, they further improved glycemic control compared to MST vegetables.

Fucoidans, sulfated polysaccharides extracted from brown algae, are marine products with the potential to modulate bone formation and vascularization processes. The bioactivity and safety of fucoidans are highly associated with their chemical structure, which may vary with algae species and extraction method. Thus, in depth evaluation of fucoidan extracts in terms of endotoxin content, cytotoxicity, and their detailed molecular biological impact on the individual cell types in bone is needed. In this study, we characterized fucoidan extracts from three different Fucus species including Fucus vesiculosus (Fv), Fucus serratus (Fs), and Fucus distichus subsp. evanescens (Fe) for their chemical features, endotoxin content, cytotoxicity, and bioactive effects on human outgrowth endothelial cells (OEC) and human mesenchymal stem cells (MSC) as in vitro models for bone function and vascularization. Extracts contained mainly high molecular weight (HMW) fucoidans and were free of endotoxins that may cause inflammation or influence vascularization. OEC tolerated fucoidan concentrations up to 200 µg/mL, and no indication of cytotoxicity was observed. The inflammatory response, however, investigated by real-time PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) and endothelial barrier assessed by impedance measurement differed for the individual extracts. MSC in comparison with endothelial cells were more sensitive to fucoidans and showed partly reduced metabolic activity and proliferation at higher doses of fucoidans. Further results for MSC indicated impaired osteogenic functions in alkaline phosphatase and calcification assays. All tested extracts consistently lowered important molecular mediators involved in angiogenesis, such a VEGF (vascular endothelial growth factor), ANG-1 (angiopoietin 1), and ANG-2 (angiopoietin 2), as indicated by RT-PCR and ELISA. This was associated with antiangiogenic effects at the functional level using selected extracts in co-culture models to mimic bone vascularization processes during bone regeneration or osteosarcoma.

Fucoidans are algal polysaccharides that exhibit protective properties against oxidative stress. The aim of this study was to investigate different fucoidans from brown seaweeds for their ability to protect against iron-dependent oxidative stress (ferroptosis), a main hallmark of retinal and brain diseases, including hemorrhage. We investigated five new high-molecular weight fucoidan extracts from Fucus vesiculosus, F. serratus, and F. distichus subsp. evanescens, a previously published Laminaria hyperborean extract, and commercially available extracts from F. vesiculosus and Undaria pinnatifida. We induced oxidative stress by glutathione depletion (erastin) and H2O2 in four retinal and neuronal cell lines as well as primary cortical neurons. Only extracts from F. serratus, F. distichus subsp. evanescens, and Laminaria hyperborea were partially protective against erastin-induced cell death in ARPE-19 and OMM-1 cells, while none of the extracts showed beneficial effects in neuronal cells. Protective fucoidans also attenuated the decrease in protein levels of the antioxidant enzyme GPX4, a key regulator of ferroptosis. This comprehensive analysis demonstrates that the antioxidant abilities of fucoidans may be cell type-specific, besides depending on the algal species and extraction method. Future studies are needed to further characterize the health-benefiting effects of fucoidans and to determine the exact mechanism underlying their antioxidative abilities.

Phycocyanins from cyanobacteria are possible sources for new natural blue colourants. Their chromophore, phycocyanobilin (PCB), was cleaved from the apoprotein by solvolysis in alcohols and alcoholic aqueous solutions. In all cases two PCB isomers were obtained, while different solvent adducts were formed upon the use of different reagents. The reaction is believed to take place via two competing pathways, a concerted E2 elimination and a S N2 nucleophilic substitution. Three cleavage methods were compared in terms of yield and purity: conventional reflux, sealed vessel heated in an oil bath, and microwave assisted reaction. The sealed vessel method is a new approach for fast cleavage of PCB from phycocyanin and gave at 120 °C the same yield within 30 min compared to 16 h by the conventional reflux method (P

In this work, sunflower oil (SF) and methyl ester of sunflower oil (ME-SF) were introduced as two green solvents for extracting astaxanthin (ASX) from shrimp processing waste. The effects of temperature (25, 45, 70 °C), solvent to waste ratio (3, 6, 9), waste particle size (0.6 and 2.5 mm) and moisture content (0% and 86.8%) as well as stirrer speed (120, 200, 400 rpm) were studied during the process. In all the experiments, ASX yields from extraction with ME-SF were higher than with SF. For both solvents, the highest ASX content using a short extraction time was achieved at a temperature of 70 °C, a solvent to waste ratio of 9 and a stirrer speed of 400 rpm while the waste particle size and moisture content were 0.6 mm and 86.8%, respectively. ME-SF and SF seem promising solvents extracting respectively about 80% and 60% of the total ASX extracted by organic solvents.

Falcarinol (FaOH) and falcarindiol (FaDOH) are found in many food plants of the Apiaceae family. Carrots are a major dietary source of these polyacetylenes. Feeding azoxymethane (AOM)-induced rats with carrots and purified FaOH have previously been shown to inhibit neoplastic transformations in the colon. FaOH and FaDOH have also shown to have a synergistic effect in vitro, resulting in a significant increased cytotoxic activity. Based on these findings the antineoplastic effect of FaOH and FaDOH (purity > 99%) was investigated in the AOM-induced rat model. Twenty rats received rat diet containing 7 μg FaOH per g feed and 7 μg FaDOH per g feed and 20 rats were controls receiving only rat diet. Then carcinogenesis was induced in all 40 rats with the carcinogen AOM. All animals received the designated diet for 2 weeks before AOM induction and continued on the designated diet throughout the experiment. Rats were euthanized 18 weeks after the first AOM injection and macroscopic polyp/cancers were measured, harvested and stained for histology. The difference in sizes of aberrant crypt foci (ACF) were analysed in a Wilcoxon rank sum test, in which the median number of small ACF was 218 in controls and 145 in polyacetylene treated rats (P < 0.001). Fifteen control rats and 8 treated rats had macroscopic tumors (P = 0.027). The number of tumors larger than 3 mm were 6 and 1 in control and treated rats, respectively (P = 0.032). In conclusion dietary supplements with FaOH and FaDOH reduced the number of neoplastic lesions as well as the growth rate of the polyps suggesting a preventive effect of FaOH and FaDOH on the development of colorectal cancer.

Sunflower oil (SF) and its methyl ester as well as supercritical fluid (SC-CO2+ 5 wt% EtOH) have recently been shown as potential green solvents which could substitute traditional organic solvents. This study investigates the economic feasibility of using these green solvents to extract astaxanthin (ASX) from shrimp processing waste. The feasibility of commercial use of the green solvents under plausible process conditions is compared to extraction with a mixture of hexane: isopropanol (Hex:IPA). The process flowsheets describing these processes were modelled by means of SuperPro Designer and Aspen Plus. The highest ASX extraction yield was achieved using Hex:IPA (26.4 kg/year), the lowest yield using SC-CO2+ 5 wt% EtOH (12.8 kg/year). The cost of producing pure ASX for these processes was about 0.6 and 0.82 $/mg of ASX, respectively. The concentration of ASX obtained by the extraction processes with SF or the methyl ester of SF (ME-SF) was 2.5 and 153 ppm with a production cost of 0.06 and 0.16 $/mg of ASX, respectively. In addition, shrimp feed production was considered as a feasible application of the low concentration ASX obtained by SF extraction. A combination of ASX extracted with SF and synthetic ASX yielded a shrimp feed production cost comparable to the current market price. The calculated feed price based on the ASX production cost of the other green processes, ME-SF and SCFE, resulted in a significantly higher production cost.

Background: Female age-related estrogen deficiency increases the risk of osteoporosis, which can be effectively treated with the use of hormone replacement therapy. However, hormone replacement therapy is demonstrated to increase cancer risk. Bioavailable isoflavones with selective estrogen receptor affinity show potential to prevent and treat osteoporosis while minimizing or eliminating carcinogenic side effects.Objective: In this study, we sought to determine the beneficial effects of a bioavailable isoflavone and probiotic treatment against postmenopausal osteopenia. Design: We used a novel red clover extract (RCE) rich in isoflavone aglycones and probiotics to concomitantly promote uptake and a favorable intestinal bacterial profile to enhance isoflavone bioavailability. This was a 12-mo, double-blind, parallel design, placebo-controlled, randomized controlled trial of 78 postmenopausal osteopenic women supplemented with calcium (1200 mg/d), magnesium (550 mg/d), and calcitriol (25 mg/d) given either RCE (60 mg isoflavone aglycones/d and probiotics) or a masked placebo [control (CON)].Results: RCE significantly attenuated bone mineral density (BMD) loss at the L2–L4 lumbar spine vertebra (P , 0.05), femoral neck (P , 0.01), and trochanter (P , 0.01) compared with CON (20.99% and 22.2%; 21.04% and 23.05%; and 20.67% and 22.79, respectively). Plasma concentrations of collagen type 1 cross-linked C-telopeptide was significantly decreased in the RCE group (P , 0.05) compared with CON (29.40% and 26.76%, respectively). RCE significantly elevated the plasma isoflavone concentration (P , 0.05), the urinary 2-hydroxyestrone (2-OH) to 16a-hydroxyestrone (16a-OH) ratio (P , 0.05), and equol-producer status (P , 0.05) compared with CON. RCE had no significant effect on other bone turnover biomarkers. Self-reported diet and physical activity were consistent and differences were nonsignificant between groups throughout the study. RCE was well tolerated with no adverse events.Conclusions: Twice daily RCE intake over 1 y potently attenuated BMD loss caused by estrogen deficiency, improved bone turnover, promoted a favorable estrogen metabolite profile (2-OH:16a-OH), and stimulated equol production in postmenopausal women with osteopenia. RCE intake combined with upplementation (calcium, magnesium, and calcitriol) was more effective than supplementation alone. This trial was registered at clinicaltrials.gov as NCT02174666.

Summary Background. Immediate hypersensitivity reactions to root vegetables of the Umbelliferae plant family (Apiaceae) is well known. Delayed-type hypersensitivity is rarely reported. Objective. To report the first case of systemic contact dermatitis caused by root vegetables and some chemical implications. Materials and methods. Prick and patch testing were performed with fresh vegetables and selected allergens, and this was followed by high-performance liquid chromatography‐mass spectrometry (MS)/MS analysis of the falcarinol syringe. Results. The patient was contact-sensitive to celeriac, parsnip, and carrot, but tested negative to falcarinol. Subsequent analysis showed that the syringe contained falcarinol. Conclusion. The non-occupational sensitization resulting from both direct and systemic contact with Apiaceae root vegetables was apparently not caused by falcarinol.

In modern society, novel marine resources are scrutinized pursuing compounds of use in the medical, pharmaceutical, biotech, food or feed industry. Few of the numerous marine macroalgae are currently exploited. In this study, the contents of nutritional compounds from nine common North Atlantic red macroalgae were compared: the lipid content was low and constant among the species, whereas the fatty acid profiles indicated that these species constitute interesting sources of polyunsaturated fatty acids (PUFA). The dominating essential and non-essential amino acids were lysine and leucine, aspartic acid, glutamic acid, and arginine, respectively. The amino acid score of the nine algae varied from 44% to 92%, the most commonly first limiting amino acid being histidine. Lutein, β-carotene, and zeaxanthin were the identified carotenoids. Contents of all macro and trace minerals, with the exception of phosphorus, were higher than those described for conventional food. Low sodium/potassium ratios (0.08 - 2.54) suggested a potential for using the ash fraction for sodium salt replacement. The algae constituted rich sources of carbohydrates (40% to 71% of DM) which show their potential for a broader commercial exploitation. In some species, the concentrations of arsenic, cadmium, and lead exceeded limit values for application in food or feed. In conclusion, the nine algae represent promising potential sources of health promoting additives for human and animal diets, in whole or in a biorefinery concept.

Sunflower oil and its methyl ester have recently been shown as potential green solvents which could substitute traditional organic solvents. This study investigates the economic feasibility of using these green solvents to extract astaxanthin from shrimp processing waste. The feasibility of commercial use of the green solvents under plausible process conditions is compared to extraction with a mixture of hexane: isopropanol. Process flowsheets describing these processes were created using SuperPro Designer. Although the astaxanthin extraction yield was highest when using hexane: isopropanol, according to the economic analysis, the green solvents showed lower capital and operating costs. Extraction with methyl ester of sunflower oil was found to be the more efficient green solvent process investigated with respect to production rate and unit cost of concentrated astaxanthin (155 ppm).

Roots of Echinacea purpurea and Echinacea pallida cultivated for 4 years in a North European climate were analyzed for seasonal variations in the concentrations of lipophilic constituents (alkamides, ketoalkenes, and ketoalkynes) and phenolic acids by harvesting five times during 1 year to establish the optimal time for harvest. A total of 16 alkamides, three ketoalkenes, two ketoalkynes, and four phenolic acids (echinacoside, cichoric acid, caftaric acid, and chlorogenic acid) were identified in aqueous ethanolic (70%) extracts by liquid chromatography-mass spectrometry and quantified by reverse-phase high-performance liquid chromatography. The major alkamides in the roots of E. purpurea were at their lowest concentration in the middle of autumn and early winter, and the total concentration of lipophilic compounds in E. pallida showed the same pattern. Moreover, all of the major phenolic acids in E. purpurea were at their highest concentrations in spring. The optimal harvest time in spring is in contrast to normal growing guidelines; hence, this specific information of seasonal variations in the concentrations of lipophilic and phenolic compounds in E. purpurea and E. pallida is valuable for research, farmers, and producers of medicinal preparations.

The concentration of the lifesaving antimalarial compound artemisinin (AN) in cultivated Artemisia annua (A. annua) plants is relatively low, and thus research in improving the content is important. In the present study, external stress was applied to adult plants of A. annua and the effect was examined on the concentrations of AN and its immediate precursors in leaves, and these concentrations were related to densities and sizes of the glandular trichomes (GT). Plants were stress treated weekly five times by sandblasting or spraying with salicylic acid, chitosan oligosaccharide, H 2O 2, and NaCl solutions. Contents of AN-related compounds (AN-c) were analysed in leaf samples from an upper and a lower position of the plants, and GT were quantified and measured. In lower leaves, several stress treatments had significant negative effects on concentrations of AN-c, whereas the ratios between compounds showed an increased conversion to AN. In the upper leaves, no changes were observed compared to controls. Linear relations were found between the concentrations of metabolites and the density of GT in both upper and lower leaves, and size of GT in lower leaves. Results suggested that older and younger leaves may respond differently to applied stress. A part of the plants were infected by powdery mildew, and this caused significantly different compositions of the AN-c, compared to uninfected plants. In conclusion, changes in concentrations of AN-c seemed largely to be related to changes in GT densities and sizes.

Greek oregano is commonly used as a spice and in traditional medicine in Eurasia. The plant is rich in secondary metabolites, such as volatile organic compounds (VOC) and polyphenols. Chitosan oligosaccharides (COS) are used as a plant elicitor. The objectives of this study were to determine the effects of COS on the growth and content of secondary metabolites in Greek oregano. Four COS treatments (50, 200, 500, and 1000 ppm) were used in a field experiment. The 200 and 500 ppm COS treatments promoted plant height growth, whereas 50 and 200 ppm COS upregulated the content of polyphenols significantly (38 and 29%, respectively). The COS treatments induced H(2)O(2) generation in Greek oregano leaves; thus, the effect of H(2)O(2) treatment was studied to investigate the possible role of H(2)O(2) in growth and polyphenol production. A low concentration of H(2)O(2) also promoted plant height growth, but only tendencies to higher polyphenol content were seen.

In a search for more effective and safe anti-diabetic compounds, we developed a pharmacophore model based on partial agonists of PPARγ. The model was used for the virtual screening of the Chinese Natural Product Database (CNPD), a library of plant-derived natural products primarily used in folk medicine. From the resulting hits, we selected methyl oleanonate, a compound found, among others, in Pistacia lentiscus var. Chia oleoresin (Chios mastic gum). The acid of methyl oleanonate, oleanonic acid, was identified as a PPARγ agonist through bioassay-guided chromatographic fractionations of Chios mastic gum fractions, whereas some other sub-fractions exhibited also biological activity towards PPARγ. The results from the present work are two-fold: on the one hand we demonstrate that the pharmacophore model we developed is able to select novel ligand scaffolds that act as PPARγ agonists; while at the same time it manifests that natural products are highly relevant for use in virtual screening-based drug discovery.

The feasibility of using a chromatography-crystallization hybrid separation process for isolation and purification of artemisinin from the herbal plant Artemisia annua was studied. After separating the crude extracts of Artemisia annua by flash column chromatography, a two-step antisolvent crystallization process was used for further isolation and purification. The two-step antisolvent crystallization process was designed based on the solubility of artemisinin in different solvents and solvent mixtures. The first step crystallization process was to remove impurities from the solution and the second step to isolate artemisinin. The crystallization behavior of the two polymorphs of artemisinin during the antisolvent crystallization process was also studied by performing crystallization in solutions of artemisinin in acetonitrile and acetone, respectively. It was observed that the formation of the two polymorphs of artemisinin in acetone solution follows the Ostwald's rule of stages. Fast feeding of the antisolvent (water) resulted in crystallization of the metastable triclinic form which then transformed to the stable orthorhombic form through a solvent-mediated transformation mechanism. The stable orthorhombic form crystallized out when water was fed slowly. Crystallization from acetonitrile always yielded the stable orthorhombic form regardless of the feeding rate of the antisolvent. Finally, a chromatography-crystallization hybrid separation process was proposed and tested. The results of the present work demonstrated the feasibility of combining the advantages of column chromatography and crystallization to improve efficiency of the isolation of artemisinin from Artemisia annua.

The effect of toasting field beans and of grass-clover: maize silage ratio on milk production, milk composition and the sensory quality of the milk was investigated in a 2 2 factorial experiment. Toasting of field beans resulted in lower milk contents of both fat (44.2 versus 46.1 g/kg, P = 0.02) and protein (33.5 versus 34.2 g/kg, P = 0.008), whereas milk production, urea and somatic cell contents were unaffected compared with the untreated field beans. Increasing the proportion of maize silage (from 9 to 21% of DM) in the ration decreased the content of urea in milk (P = 0.002), whereas milk production and milk content of fat and protein were unaffected. Milk from cows fed the high proportion of maize silage had a lower (P = 0.04) long-chain fatty acid content (≥ C18). Furthermore, milk from cows fed the high proportion of maize silage had a lower (13-26%) content of luteine (P = 0.03), 13-cis-β-carotene (P = 0.04) and β-carotene (P = 0.05). Toasting of field beans compared with untreated field beans did not affect the milk content of carotenoids and had only small effects on fatty acid composition. Regarding the sensory quality, the four treatments resulted in milk being characterized by a distinctly fatty mouthfeel and creamy flavour and a pronounced sugar-sweet taste and creamy odour. The higher proportions of maize in the feed resulted, in general, in milk characterized by a significantly more intense maize odour (P P

A detailed phytochemical investigation of a dichloromethane extract of the resinous exudates of the cushion bush plant (Leucophyta brownii) resulted in the isolation of the new 8,12-guaianolides leucophytalins A (5) and B (6), the new 1,10-seco-eudesmane leucophytalin C (10), six rare 8,12-guaianolides (1–4, 7, and 8), and the xanthanolide tomentosin (9). The structures of all isolated compounds were elucidated on the basis of spectroscopic and spectrometric analyses. The structures of compounds isolated in crystalline form, including leucophytalins A and C, were further confirmed by X-ray crystallography. The crude extract exhibited moderate cytostatic activity against a breast cancer (MCF-7) and human colon cancer (HT-29) cell line with IC50 values of 9.3 and 18 μg/mL, respectively, and anti-inflammatory activity against the macrophage-like cell line RAW 264.7 with IC50 values of 3.9 and 6.1 μg/mL for thromboxane B2 and prostaglandin E2 production, respectively. The isolated compounds were evaluated for their cytostatic activity against MCF-7 and HT-29 cells (1, 3–10) and their anti-inflammatory activity against RAW 264.7 cells (1–10). All isolated compounds are most likely derived from (+)-germacrene A, and a biosynthetic pathway is proposed for these sesquiterpenoids.

We examined the effect of light emitting diode (LED) lighting in greenhouse facilities on growth, chlorophyll fluorescence and pigmentation in Phalaenopsis 'Vivien' and 'Purple Star' under purpose-built LED arrays yielding c. 200 µmol m(-2) s(-1) at plant height for 14 h per day and 24/18°C day/night temperature, respectively, from January to April 2013. The light treatments were (1) 40% blue in 60% red (40% B/R), (2) 0% blue in 100% red (0% B/R) and (3) white LEDs with 32% blue in white (32% B/W, control), with background daylight under shade screens. The plants were harvested twice for leaf growth and pigmentation. There was no clear pattern in the spectral effect on growth since the order of leaf size differed between harvests in March and April. Fv /Fm was in the range of 0.52-0.72, but overall slightly higher in the control, which indicated a permanent downregulation of PSII in the colored treatments. The fluorescence quenching showed no acclimation to color in 'Purple Star', while 'Vivien' had lower ETR and higher NPQ in the 40% B/R, resembling low light acclimation. The pigmentation showed corresponding spectral response with increasing concentration of lutein while increasing the fraction of blue light, which increased the light absorption in the green/yellow part of the spectrum. The permanent downregulation of PSII moved a substantial part of the thermal dissipation from the light regulated NPQ to non-regulated energy losses estimated by ΦNPQ and ΦNO and the difference found in the balance between ΦPSII and ΦNPQ in 'Vivien' disappeared when ΦNO was included in the thermal dissipation.

Scope: Omega-6 (n-6) PUFA-rich diets are generally considered obesogenic in rodents. Here, we examined how long-term intake of a high-fat/high-sucrose (HF/HS) diet based on safflower oil affected metabolism, inflammation, and gut microbiota composition. Methods and results: We fed male C57BL/6J mice a HF/HS diet based on safflower oil—rich in n-6 PUFAs—or a low-fat/low-sucrose diet for 40 wk. Compared to the low-fat/low-sucrose diet, intake of the safflower-based HF/HS diet only led to moderate weight gain, while glucose intolerance developed at week 5 prior to signs of inflammation, but concurrent with increased levels of linoleic acid and arachidonic acid in hepatic phospholipids. Intake of the HF/HS diet resulted in early changes in the gut microbiota, including an increased abundance of Blautia, while late changes coincided with altered inflammatory profiles and increased fasting plasma insulin. Analysis of immune cells in visceral fat and liver revealed no differences between diets before week 40, where the number of immune cells decreased in the liver of HF/HS-fed mice. Conclusion: We suggest that a diet-dependent increase in the n-6 to omega-3 (n-3) PUFA ratio in hepatic phospholipids together with gut microbiota changes contributed to early development of glucose intolerance without signs of inflammation.

To investigate the effect of the light spectrum on photosynthesis, growth, and secondary metabolites Rosa hybrida 'Scarlet', Chrysanthemum morifolium 'Coral Charm', and Campanula portenschlagiana 'BluOne' were grown at 24/18°C day/night temperature under purpose-built LED arrays yielding approximately 200 μmol m(-2)s(-1) at plant height for 16 h per day. The four light treatments were (1) 40% Blue/60% Red, (2) 20% Blue/80% Red, (3) 100% Red, and (4) 100% White (Control). The plant height was smallest in 40% Blue/60% Red in roses and chrysanthemums, while the biomass was smallest in the white control in roses and in 100% Red in chrysanthemums. The total biomass was unaffected by the spectrum in campanulas, while the leaf area was smallest in the 40% Blue/60% Red treatment. In 100% Red curled leaves and other morphological abnormalities were observed. Increasing the blue to red ratio increased the stomatal conductance though net photosynthesis was unaffected, indicating excess stomatal conductance in some treatments. With higher blue light ratio all phenolic acids and flavonoids increased. In view of the roles of these secondary metabolites as antioxidants, anti-pathogens, and light protectants, we hypothesize that blue light may predispose plants to better cope with stress.

Clostridium perfringens is the causative agent of necrotic enteritis leading to significant losses in the poultry industry. Dichloromethane and n-hexane extracts of aerial parts of Artemisia annua (Asteraceae) exhibited activity against C. perfringens with minimum inhibitory concentrations (MIC) of 185 and 270 μg/mL, respectively. Bioassay-guided fractionation of the extracts gave several active fractions (MIC between 75 and 600 μg/mL). Investigations of the most active fractions resulted in the iso ponticaepoxide (MIC between 100 and 200 μg/mL) and (+)-threo-(5E)-trideca-1,5-dien-7,9,11-triyne-3,4-diol (MIC between 400 and 800 μg/mL), the flavonols chrysosplenol D (MIC between 200 and 400 μg/mL) and casticin (slight inhibition at 800 μg/mL), and 2,4-dihydroxy-6-methoxyacetophenone (slight inhibition at 800 μg/mL). Also, the coumarin scopoletin and the sesquiterpene lactone artemisinin were isolated from active fractions but showed no inhibition of C. perfringens growth at 800 and 2000 μg/mL, respectively. Fractions containing essential oil components with camphor constituting >60% did not show inhibition of C. perfringens up to 1600 μg/mL. Extracts and some active fractions showed higher antibacterial effect than individual bioactive compounds, suggesting that synergistic effects may underlie the observed antibacterial effect. The present study adds new valuable information on the antibacterial effect of A. annua against C. perfringens.

Echinacea purpurea has been used in traditional medicine as a remedy for the treatment and pre- vention of upper respiratory tract infections and the common cold. Recent investigations have in- dicated that E. purpurea also has an effect on insu- lin resistance. A dichloromethane extract of E. purpurea roots was found to enhance glucose up- take in adipocytes and to activate peroxisome proliferator-activated receptor γ. The purpose of the present study was to identify the bioactive compounds responsible for the potential antidia- betic effect of the dichloromethane extract using a bioassay-guided fractionation approach. Basal and insulin-dependent glucose uptake in 3T3-L1 adipocytes were used to assess the bioactivity of extract, fractions and isolated metabolites. A per- oxisome proliferator-activated receptor γ transac- tivation assay was used to determine the peroxi- some proliferator-activated receptor γ activating properties of the extract, active fractions and iso- lated metabolites. Two novel isomeric dodeca- 2E,4E,8Z,10E/Z-tetraenoic acid 2-methylbutyl- amides together with two known C12-alkamides and α-linolenic acid were isolated from the active fractions. The isomeric C12-alkamides were found to activate peroxisome proliferator-activated re- ceptor γ, to increase basal and insulin-dependent glucose uptake in adipocytes in a dose-dependent manner, and to exhibit characteristics of a perox- isome proliferator-activated receptor γ partial ag- " Echinacea purpurea l " Asteraceae l " alkamides

Full-grown Artemisia annua plants were subjected to chemical and physical stress conditions, and the effect of these on the concentration and chemical composition of essential oil components (EOC) in the leaves was studied. The chemical stress treatments were performed by foliar application of NaCl, H 2O 2, salicylic acid and chitosan oligosaccharide (COS). The EOC of the leaves were extracted with n-hexane and identified and quantified by GC-MS and GC-FID, respectively. Approximately 96 % of EOC in the extracts were identified and quantified of which β-pinene, camphene, germacrene D, camphor, coumarin and dihydro-epi-deoxyarteannuin B were the major EOC accounting for about 75 % of the total content of EOC in the extracts. The physical stress treatment, sandblasting of the plants resulted in a significant enhancement in the content of α-pinene, camphene, coumarin and dihydro-epi-deoxyarteannuin B. The total yield of identified EOC in non-treated plants (control) was 86.2 ± 13.8 μg g -1 fresh weight (FW) compared with 104.0 ± 9.1 μg g -1 FW in sandblasted plants. The chemical stress treatments did not affect the composition of EOC significantly. The results indicate that chemical stress treatments do not affect the concentration and composition of EOC in full-grown A. annua plants to the same extent as physical stress treatment by sandblasting.

I. Abstract : Based on a bio-assay guided fractio~tio~, a new sterol has been isolated as an equilibrium of isomers 1 with interesting cancer cell antiproliferative properties from a yet undcscribcd species of the. genus Ctemcellu sp. The structure of this sterol has been inferred f" the acetyl derivative 2, on the basis of spectral shldy. Copyright Q 1996 Elsevier Science Ltd Gorgonians are a rich source of biologically active products among which are prostaglandins3.4, diterpenes3.5-7 (mainly briaranes and cembrenes), polymethylene butenolides8 and sterolsg. Several compounds have shown a broad range of potent bioactivities, mainly antimicrobiallo, anticancer11.12. and antiinfla"atory413. Ctenocella belongs to the little investigated Ellisellidae family (Cnidaria) which yielded usual sterols (Ellisella paraplexauroia'esg), juncins A-E (Junceella juncea14), junceellolides A-D (Junceella fragilisl3), junceellin and praelolide (Junceella squamata13), gemacolides A-E (Junceella gemmacea15) while

The aerial parts of the plant Artemisia annua contain essential oils having antimicrobial properties against Clostridium perfringens Type A, the causal agent for necrotic enteritis in broilers. In two experiments, the influence of increasing dietary concentrations of dried A. annua leaves (0, 5, 10 and 20 g/kg) and n-hexane extract from fresh A. annua leaves (0, 125, 250 and 500 mg/kg) on broiler performance was investigated. Dried plant material decreased feed intake and body weight in a dose-dependent manner, and 10 and 20 g/kg diet tended to improve the feed conversion ratio. The n-hexane extract also reduced feed intake, but broiler weight tended to decrease only at the highest dietary concentration. The feed conversion ratio tended to improve when birds received 250 and 500 mg/kg n-hexane extract. In a third experiment, a necrotic enteritis disease model was applied to investigate the effect of the dietary addition of dried A. annua leaves (10 g/kg on top) or n-hexane extract of A. annua (250 mg/kg) on the severity of the disease in broilers. The addition of n-hexane extract reduced the intestinal C. perfringens numbers and the severity of the disease-related small intestinal lesions. Over the infection period from day 17 to day 27, birds supplemented with the n-hexane extract gained more weight than both the challenged control birds and birds receiving dried plant material. The results indicate that n-hexane extracts derived from A. annua can modulate the course of necrotic enteritis and compensate to a certain extent for the disease-associated weight losses.

Five different Artemisia annua-derived materials (i.e. dry leaves, pure artemisinin, and hexane, dichloromethane or methanol extracts of leaves) were screened for their in vitro activities against six clonal cultures of Histomonas meleagridis. Except for the methanol extract, all tested materials displayed in vitro activity against all tested protozoal clones. Neither the dry plant material, extracts nor artemisinin showed any antibacterial activity against the xenic bacteria accompanying the six H. meleagridis clones at concentration levels identical to the antihistomonal setting. The dichloromethane extract of dry leaves (Ext-DCM) (minimal lethal concentration=1.0 mg/ml) and artemisinin (half-maximal inhibitory concentration=1.295 mg/ml) had the most promising antihistomonal properties and were therefore subsequently tested in a standardized experimental infection model in both turkeys and chickens infected with clonal H. meleagridis. There were no differences between treatment groups, where all infected turkeys showed severe clinical histomonosis and demonstrated severe typhlohepatitis typical for histomonosis. Consistent with the infection model used, the infected chickens did not show any adverse clinical signs but contracted severe lesions in their caeca 7 and 10 days post infection (d.p.i.), liver lesions were absent to mild after 7 d.p.i. and progressed to severe lesions at 10 d.p.i.; thus no differences between treatment groups were observed. In conclusion, neither artemisinin nor Ext-DCM was able to prevent experimental histomonosis in turkeys and chickens at the given concentrations, which is contrary to the antihistomonal effect noticed in vitro even though the same clonal culture was used. The results of this study therefore clearly demonstrate the importance of defined in vivo experimentation in order to assess and verify in vitro results.

BACKGROUND: The aim of the present work was to study to what extent grazing large amounts of white clover (WCL), red clover (RCL), lucerne (LUC) or chicory (CHI) was suitable for production of bovine milk with a high milk fat content of tocopherols, carotenoids, α-linolenic acid and conjugated linoleic acid. RESULTS: WCL, RCL, LUC and CHI swards were established to investigate the influence of herbage composition on the milk quality of grazing dairy cows at three periods. Average milk fat composition showed high concentrations of linolenic acid (12 mg g−1), conjugated linoleic acid (13 mg g−1), carotenoids (6 µg g−1) and α-tocopherol (21 µg g−1 milk fat). There were minor differences between herbage types and periods, but multivariate analysis of these data showed no clear grouping. Chemical composition of herbage varied with species as well as period, but it was not possible to relate milk and feed contents of specific fatty acids, carotenoids or tocopherols. CONCLUSION: All four herbages tested were suitable for production of milk with a high content of beneficial compounds. Thus any of these herbages could be used in production of such differentiated milk based on a large proportion of grazing in the ration. Copyright © 2012 Society of Chemical Industry

Application of 3.6 mm silicon (Si+) to the rose (Rosa hybrida) cultivar Smart increased the concentration of antimicrobial phenolic acids and flavonoids in response to infection by rose powdery mildew (Podosphaera pannosa). Simultaneously, the expression of genes coding for key enzymes in the phenylpropanoid pathway (phenylalanine ammonia lyase, cinnamyl alcohol dehydrogenase, and chalcone synthase) was up-regulated. The increase in phenolic compounds correlated with a 46% reduction in disease severity compared with inoculated leaves without Si application (Si−). Furthermore, Si application without pathogen inoculation induced gene expression and primed the accumulation of several phenolics compared with the uninoculated Si− control. Chlorogenic acid was the phenolic acid detected in the highest concentration, with an increase of more than 80% in Si+ inoculated compared with Si− uninoculated plants. Among the quantified flavonoids, rutin and quercitrin were detected in the highest concentrations, and the rutin concentration increased more than 20-fold in Si+ inoculated compared with Si− uninoculated plants. Both rutin and chlorogenic acid had antimicrobial effects on P. pannosa, evidenced by reduced conidial germination and appressorium formation of the pathogen, both after spray application and infiltration into leaves. The application of rutin and chlorogenic acid reduced powdery mildew severity by 40% to 50%, and observation of an effect after leaf infiltration indicated that these two phenolics can be transported to the epidermal surface. In conclusion, we provide evidence that Si plays an active role in disease reduction in rose by inducing the production of antifungal phenolic metabolites as a response to powdery mildew infection.

Udgivelsesdato: 2010-Sep Background: The Compositae plant feverfew (Tanacetum parthenium) has long been recognized as an important sensitizer in European Compositae-allergic patients, mainly because of its content of the sesquiterpene lactone parthenolide. Recently, a parthenolide-depleted feverfew extract with claimed anti-inflammatory properties has been developed for use in cosmetics. Objective: The aim of the study was to test, on the basis of patch test reactions, whether persons with contact allergy to feverfew could tolerate creams containing this feverfew extract. Patients/Materials/Methods: Seven patients with feverfew contact allergy were patch tested with two creams containing the feverfew extract. Subsequently, the creams were analysed by liquid chromatography with tandem mass spectrometry to detect parthenolide. Results: Four of the patients tested positive to one of the creams; reactivity was associated with simultaneous positive reactions to parthenolide. This cream was analysed about 2 years later, and no parthenolide was detected, probably because of degradation of the compound. Conclusions: Topical products containing parthenolide-depleted feverfew extracts may elicit positive patch test reactions in feverfew-sensitive patients. The reactivity may be enhanced by simultaneous testing with parthenolide, but the reactivity is lost over time, probably because of degradation of parthenolide.

In this work, a hybrid chromatography−crystallization separation process is suggested and is used to isolate and purify the antimalarial drug artemisinin from the Chinese medicinal herb Artemisia annua. Extraction of the aerial parts of A. annua by an organic solvent yielded a complex mixture of natural products. A hybrid separation process was therefore proposed to pursue higher recovery and purity of artemisinin. The crude extract was first separated by flash column chromatography, and the fractions containing artemisinin were then separated with a two-step antisolvent crystallization process to isolate artemisinin of high purity. The antisolvent crystallization process was designed on the basis of the solubility of artemisinin in different solvents and the crystallization behaviour of the two polymorphs of artemisinin. The first step in the crystallization process was used to remove impurities from the solution, and the second step was used to isolate the stable orthorhombic form of artemisinin. The re...