Natural Products from Leaves of the Ancient Iranian Medicinal Plant Echium amoenum Fisch. & C. A. Mey.

For several millennia, leaves of Echium amoenum Fisch. & C. A. Mey., an important Iranian medicinal plant with nutritional value as nutraceutical, have been used as tea for the treatment of several conditions, including inflammation. The nutritional value of intake of E. amoenum tea has mainly been correlated to its rich content of mainly water-soluble antioxidants. Although the entire plant is utilized, only natural products of the flowers have previously been thoroughly investigated. The rare natural products bis(3-(3,4-dihydroxyphenyl)-1-methoxy-1-oxopropan-2-yl)-1-(3,4-dihydroxyphenyl)-6,7-dihydroxy-1,2-dihydronaphthalene-2,3-dicarboxylate, 4-Oxy-(E)-caffeoyl-2,3-dihydroxybutanoic acid methyl ester and 4-Oxy-(Z)-caffeoyl-2,3-dihydroxybutanoic acid methyl ester, in addition to the widely distributed compounds rosmarinic acid methyl ester and (E)-caffeic acid, were purified and characterized from leaves of Echium amoenum. The structures were determined by a combination of several 2D NMR spectroscopic techniques, circular dichroism spectroscopy and high-resolution mass spectrometry. The fact that bis(3-(3,4-dihydroxyphenyl)-1-methoxy-1-oxopropan-2-yl)-1-(3,4-dihydroxyphenyl)-6,7-dihydroxy-1,2-dihydronaphthalene-2,3-dicarboxylate belongs to a rare group of natural products which have previously been patented for their significant anti-inflammatory activity may rationalize the traditional treatment of inflammations with E. amoenum.

Echiumin E, an Aryl Dihydronaphthalene Lignan from the Australian Invasive Plant Paterson's Curse (Echium plantagineum).

A new aryl dihydronaphthalene lignan, echiumin E (1), and four known compounds, echiumin A, globoidnan A, (-)-rabdosiin, and rosmarinic acid (2-5), were isolated from the Australian invasive plant Echium plantagineum (Paterson's curse) for the first time. Echiumin E (1) was characterized by 1D/2D NMR spectroscopy and MS spectrometry, with its absolute configuration assigned through comparison of experimental and TDDFT-calculated ECD data. Echiumin E (1) along with compounds 3-5 were screened in vitro against three cancer cell lines (SH-SY5Y, HeLa, and PC-3) and a prostate stromal (normal) cell line (WPMY-1) using a resazurin reduction assay. Echiumin E (1) was found to be active toward HeLa cells (IC50 0.21 µM).

Isolation of Echimidine and Its C-7 Isomers from Echium plantagineum L. and Their Hepatotoxic Effect on Rat Hepatocytes.

Echimidine is the main pyrrolizidine alkaloid of Echium plantagineum L., a plant domesticated in many countries. Because of echimidine's toxicity, this alkaloid has become a target of the European Food Safety Authority regulations, especially in regard to honey contamination. In this study, we determined by NMR spectroscopy that the main HPLC peak purified from zinc reduced plant extract with an MS [M + H]+ signal at m/z 398 corresponding to echimidine (1), and in fact also represents an isomeric echihumiline (2). A third isomer present in the smallest amount and barely resolved by HPLC from co-eluting (1) and (2) was identified as hydroxymyoscorpine (3). Before the zinc reduction, alkaloids (1) and (2) were present mostly (90%) in the form of an N-oxide, which formed a single peak in HPLC. This is the first report of finding echihumiline and hydroxymyoscorpine in E. plantagineum. Retroanalysis of our samples of E. plantagineum collected in New Zealand, Argentina and the USA confirmed similar co-occurrence of the three isomeric alkaloids. In rat hepatocyte primary culture cells, the alkaloids at 3 to 300 µg/mL caused concentration-dependent inhibition of hepatocyte viability with mean IC50 values ranging from 9.26 to 14.14 µg/mL. Our discovery revealed that under standard HPLC acidic conditions, echimidine co-elutes with its isomers, echihumiline and to a lesser degree with hydroxymyoscorpine, obscuring real alkaloidal composition, which may have implications for human toxicity.

Evaluation of multilayer perceptron neural networks and adaptive neuro-fuzzy inference systems for the mass transfer modeling of Echium amoenum Fisch. & C. A. Mey.

BACKGROUND: Multilayer perceptron (MLP) feed-forward artificial neural networks (ANN) and first-order Takagi-Sugeno-type adaptive neuro-fuzzy inference systems (ANFIS) are utilized to model the fluidized bed-drying process of Echium amoenum Fisch. & C. A. Mey. The moisture ratio evolution is calculated based on the drying temperature, airflow velocity and process time. Different ANN topologies are examined by evaluating the number of neurons (3 to 20), the activation functions and the addition of a second hidden layer. Different numbers (2 to 5) and shapes of membership functions are examined for the ANFIS, using the grid partitioning method. The models with the best performance in terms of prediction accuracy, as evaluated by the statistical indices, are compared with the best fit thin-layer model and the available data from the experimental cases of 40 °C, 50 °C and 60 °C temperatures at 0.5, 0.75 and 1 ms-1 airflow velocity. RESULTS: The best performed ANFIS model, comprised by 5-2-2 of π-shaped andtriangular membership functions for time, temperature and airflow velocityinputs respectively, was able to describe the moisture ratio evolution of E. amoenum more precisely than the best ANN topology, achieving higher values of coefficientof determination (R2 ), root mean square error (RMSE) and sum of squared errors(SSE). The best thin-layer model involving six adjustable parameters, managedto describe experimental data most accurately with R2 = 0.9996, RMSE = 0.0057and SSE = 7.3·10-4 . CONCLUSION: The results of the comparative study indicate that empirical regression models with increased numbers of adjustable parameters, constitute a simpler and more accurate modeling approach for estimating the moisture ratio of E. amoenum Fisch. & C. A. Mey under fluidized bed drying. © 2021 Society of Chemical Industry.

The role of hippocampal GABAA receptors on anxiolytic effects of Echium amoenum extract in a mice model of restraint stress.

Echium amoenum (EA), a popular medicinal plant in Persian medicine, has anxiolytic, antioxidant, sedative, and anti-inflammatory effects. This study examined whether GABA-ergic signaling is involved in the anxiolytic effects of EA in mice. Sixty BALB/c mice (25-30 g) were divided into six groups (n = 10) as follows: the (I) control group received 10 ml/kg normal saline (NS). In the stress groups, the animals underwent 14 consecutive days of restraint stress (RS), and received following treatments simultaneously; (II) RS + NS; (III) RS + Diaz (Diazepam); (IV) RS + EA; (V) RS + Flu (Flumazenil) + EA; (VI) RS + Flu + Diaz. Behavioral tests including the open field test (OFT) and elevated plus maze (EPM) were performed to evaluate anxiety-like behaviors and the effects of the regimens. The plasma level of corticosterone and the hippocampal protein expressions of IL-1ß, TNF-α, CREB, and BDNF, as well as p-GABAA/GABAA ratio, were also assessed. The findings revealed that chronic administration of EA alone produced anxiolytic effects in both behavioral tests, while diazepam alone or in combination with Flu failed to decrease the anxiety-like behaviors. Furthermore, the p-GABAA/GABAA and p-CREB/CREB ratios, and protein levels of BDNF were significantly increased in the EA-received group. On the other hand, plasma corticosterone levels and the hippocampal IL-1ß and TNF-α levels were significantly decreased by EA. However, pre-treatment with GABAA receptors (GABAA Rs) antagonist, Flu, reversed the anxiolytic and molecular effects of EA in the RS-subjected animals. Our findings confirmed that alternation of GABAAR is involved in the effects of EA against RS-induced anxiety-like behaviors, HPA axis activation, and neuroinflammation.

Partial replacement of pork fat by Echium oil in reduced sodium bologna sausages: technological, nutritional and stability implications.

BACKGROUND: The present research aimed to evaluate the nutritional, technological, microstructural and sensory characteristics of bologna sausages made with pork fat replaced with Echium oil. Three different treatments were processed, all of them with approximately 35% less sodium than a regular bologna-type sausage: Control (only pork backfat addition) and T25 and T50 (replacement of 25 and 50% of the added animal fat with Echium oil respectively). RESULTS: Proximate composition, texture profile analysis, fatty acid profile and microstructure were evaluated to characterize the products. In refrigerated storage, the following characteristics were assessed: thiobarbituric acid-reactive substances (TBARS) index, objective color, pH value, microbiological counts and sensory acceptance. Both samples with Echium oil (T25 and T50) showed a healthier lipid profile, with high omega-3 content, mainly from α-linolenic and stearidonic fatty acids, consequently presenting better (P ≤ 0.05) values of nutritional indices than the Control. However, the replacement of 50% animal fat with Echium oil negatively affected (P ≤ 0.05) most of the technological characteristics and the sensory acceptance of bolognas. In contrast, the treatment with 25% replacement did not show significant differences in most of the evaluated parameters compared with the Control sample. CONCLUSION: Thus the replacement of 25% of pork backfat with Echium oil in bologna sausages can be recommended, representing a good balance between nutritional gains and sensory impairment. © 2019 Society of Chemical Industry.

Development of a Selective Adsorbing Material for Binding of Pyrrolizidine Alkaloids in Herbal Extracts, Based on Molecular Group Imprinting.

Pyrrolizidine alkaloids are secondary plant constituents that became a subject of public concern because of their hepatotoxic, pneumotoxic, genotoxic, and cytotoxic effects. Due to disregardful harvesting and/or contamination with pyrrolizidine alkaloid-containing plants, there is a high risk of ingesting these substances with plant extracts or natural products. The limit for the daily intake was set to 0.007 µg/kg body weight. If contained in an extract, cleanup methods may help to minimize the pyrrolizidine alkaloid concentration. For this purpose, a material for depleting pyrrolizidine alkaloids in herbal preparations was developed based on the approach of molecular imprinting using monocrotaline. Molecular imprinted polymers are substances with specific binding characteristics, depending on the template used for imprinting. By means of group imprinting, only one molecule is used for creating selective cavities for many molecular pyrrolizidine alkaloid variations. Design of Experiment was used for the development using a 25 screening plan resulting in 64 polymers (32 MIPs/32 NIPs). Rebinding trials revealed that the developed material can compete with common cation exchangers and is more suitable for depleting pyrrolizidine alkaloids than C18- material. Matrix trials using an extract from Chelidonium majus show that there is sufficient binding capacity for pyrrolizidine alkaloids (80%), but the material is lacking in selectivity towards pyrrolizidine alkaloids in the presence of other alkaloids with similar functional groups such as berberine, chelidonine, and coptisine. Beyond this interaction, the selectivity could be proven for other structurally different compounds on the example of chelidonic acid.

Pyrrolizidine Alkaloids: The Botanical Origin of Pollen Collected during the Flowering Period of Echium vulgare and the Stability of Pyrrolizidine Alkaloids in Bee Bread.

Previous studies have shown that pollen products sold as nutritional supplements and used in apitherapy may contain toxic pyrrolizidine alkaloids (PAs) if bees collect pollen from PA-containing plants, such as Echium vulgare. In this study, the botanical origin of pollen from two observation sites was studied. Despite a high PA content in pollen samples that bees collected during E. vulgare's flowering period, bees were found to collect relatively few Echium pollen loads. Thus, the monitoring of pollen loads collected at the apiaries is unviable to estimate the risk of PA contamination in pollen or bee bread. In a second step, the stability of PAs in bee bread samples containing PAs at concentrations of 2538 ng/g and 98 ng/g was assessed over a period of five or six months, respectively. No significant PA reduction was observed in bee bread stored at 15 °C, but there were overall PA reductions of 39% and 33% in bee bread stored at 30 °C, reflecting hive conditions. While PA N-oxides decreased over time, other types of PAs remained relatively stable. Monitoring PAs in pollen products remains important to ensure consumer safety and should include echivulgarine (and its N-oxide), the major PA type found in pollen from E. vulgare.

Nursing protects honeybee larvae from secondary metabolites of pollen.

The pollen of many plants contains toxic secondary compounds, sometimes in concentrations higher than those found in the flowers or leaves. The ecological significance of these compounds remains unclear, and their impact on bees is largely unexplored. Here, we studied the impact of pyrrolizidine alkaloids (PAs) found in the pollen of Echium vulgare on honeybee adults and larvae. Echimidine, a PA present in E. vulgare pollen, was isolated and added to the honeybee diets in order to perform toxicity bioassays. While adult bees showed relatively high tolerance to PAs, larvae were much more sensitive. In contrast to other bees, the honeybee larval diet typically contains only traces of pollen and consists predominantly of hypopharyngeal and mandibular secretions produced by nurse bees, which feed on large quantities of pollen-containing bee bread. We quantified the transfer of PAs to nursing secretions produced by bees that had previously consumed bee bread supplemented with PAs. The PA concentration in these secretions was reduced by three orders of magnitude as compared to the PA content in the nurse diet and was well below the toxicity threshold for larvae. Our results suggest that larval nursing protects honeybee larvae from the toxic effect of secondary metabolites of pollen.

Phytochemical investigation and biological activities of Echium arenarium (Guss) extracts.

The present work was developed to evaluate the in vitro antioxidant, antibacterial, antileishmanial and cytotoxic activities of Echium arenarium (Guss) extracts, and to analyze their phytochemical composition. The highest content of total phenolic compounds was obtained in the ethyl acetate extract which showed the best DPPH scavenging activity and ß-carotene bleaching inhibition (IC50 = 1.1 and 9.94 µg/mL respectively). It also exhibited the highest antibacterial activity against Gram-positive bacteria (L. monocytogenes; S. aureus; MRSA, E. faecalis and B. cereus) and antileishmanial activity against L. major (IC50 = 13.91 ±â€¯0.43 µg/mL) and L. infantum (IC50 = 9.91 ±â€¯0.15 µg/mL). Moreover, the active extract exhibited potent antiamastigote activity (IC50 = 22.48 ±â€¯0.14 µg/mL and 18.59 ±â€¯0.09 µg/mL against L. major and L. infantum respectively). Cytotoxicity studies revealed low toxicity against Raw 264.7 macrophage cell line (IC50 = 145.80 ±â€¯0.84 µg/mL, SI < 10). Luteolin-7-O-glucoside was identified as the major flavonoid component by RP-HPLC analysis. In conclusion, Echium arenarium (Guss) extract was characterized by a wide range of biological activities and could be used as a potential natural anti-infectious drug.

The use of factorial design to evaluate the oxidation of oils containing different types of omega-3 fatty acids.

BACKGROUND: Omega-3 fatty acids (n3 FA) promote beneficial effects on cardiovascular diseases, but they are highly susceptible to oxidation, which leads to the formation of potentially toxic secondary products. In this study, the antioxidant capacity of natural compounds (sinapic acid and rutin) was evaluated using an accelerated model to oxidize the oils. Five inducers (temperature, Fe2+ , 2,2'-azobis(2-amidinopropane) dihydrochloride, ascorbyl palmitate and the hydrophobic 2,2'-azobis-2,4-dimethylvaleronitrile (AMVN)) were combined in a factorial design to accelerate the oxidation of three oils (flaxseed, echium and fish) containing different sources of n3 FA. RESULTS: Lipid hydroperoxide (LOOH) and thiobarbituric acid reactive substance (TBARS) concentrations estimated using the regression models did not differ from the observed values. AMVN + Fe2+ increased TBARS in all samples. CONCLUSION: The values for the oxidative markers obtained 48 h after induction were similar to those obtained when the oils were heated at 60 ° C for 15 days. Of a number of volatile compounds formed from the oxidation of different n3 FA sources, (E,E)-2,4-heptadienal, (E,E)-2,4-decadienal, decanal, undecanal and (E)-2-undecenal were identified in all samples and could be used as more specific oxidative markers. Using the accelerated model, rutin improved the oxidative stability of fish oil, probably due to the presence of a catechol group in its chemical structure. © 2017 Society of Chemical Industry.

Metabolic Profiling and Identification of Shikonins in Root Periderm of Two Invasive Echium spp. Weeds in Australia.

Metabolic profiling can be successfully implemented to analyse a living system's response to environmental conditions by providing critical information on an organism's physiological state at a particular point in time and allowing for both quantitative and qualitative assessment of a specific subset(s) of key metabolites. Shikonins are highly reactive chemicals that affect various cell signalling pathways and possess antifungal, antibacterial and allelopathic activity. Based on previous bioassay results, bioactive shikonins, are likely to play important roles in the regulation of rhizosphere interactions with neighbouring plants, microbes and herbivores. An effective platform allowing for rapid identification and accurate profiling of numerous structurally similar, difficult-to-separate bioactive isohexenylnaphthazarins (shikonins) was developed using UHPLC Q-TOF MS. Root periderm tissues of the invasive Australian weeds Echium plantagineum and its congener E. vulgare were extracted overnight in ethanol for shikonin profiling. Shikonin production was evaluated at seedling, rosette and flowering stages. Five populations of each species were compared for qualitative and quantitative differences in shikonin formation. Each species showed little populational variation in qualitative shikonin production; however, shikonin was considerably low in one population of E. plantagineum from Western New South Wales. Seedlings of all populations produced the bioactive metabolite acetylshikonin and production was upregulated over time. Mature plants of both species produced significantly higher total levels of shikonins and isovalerylshikonin > dimethylacrylshikonin > shikonin > acetylshikonin in mature E. plantagineum. Although qualitative metabolic profiles in both Echium spp. were nearly identical, shikonin abundance in mature plant periderm was approximately 2.5 times higher in perennial E. vulgare extracts in comparison to those of the annual E. plantagineum. These findings contribute to our understanding of the biosynthesis of shikonins in roots of two related invasive plants and their expression in relation to plant phenological stage.

A two-week, double-blind, placebo-controlled trial of Viola odorata, Echium amoenum and Physalis alkekengi mixture in symptomatic benign prostate hyperplasia (BPH) men.

CONTEXT: As an alternative approach, administration of phytotherapeutic agents in management of benign prostate hyperplasia (BPH), is rapidly growing each day. Different authors have indicated effectiveness of Viola odorata L. (Violaceae), Echium amoenum Fisch. & C.A.Mey. (Boraginaceae) and Physalis alkekengi L. (Solanaceae) in treatment of BPH. However, none have reported the beneficial outcomes of the mixture yet. OBJECTIVE: This study evaluates the therapeutical effects of V. odorata, E. amoenum and P. alkekengi mixture on symptomatic BPH patients. MATERIALS AND METHODS: Eighty six symptomatic BPH patients with International Prostate Symptom Score (IPSS) of more than 13 and prostate volume of more than 30 cm3 were randomly allocated to receive a two-week course of placebo (control group) or 1 mL of mixed hydro-alcoholic solution of P. alkekengi, E. amoenum and V. odorata extracts (1.5, 1 and 1.5% respectively) (treatment group). RESULTS: IPSS score of incomplete urination (42.3 ± 2.04%), frequency of urination (20.08 ± 1.02%), intermittency (40.78 ± 2.16%), urgency (60.91 ± 3.14%), weak stream (50.58 ± 2.14%), straining (55.67 ± 2.53%) and nocturia (40.14 ± 1.89%) in treatment group were significantly decreased after treatment compare to placebo receiving group. Furthermore, the prostate volume (16.92 ± 0.89%) and extant urine volume (28.12 ± 1.36%) also significantly decreased in treatment group compared to control group. No significant side effects or abnormalities in biochemical tests and urinalysis were observed throughout the study. DISCUSSION AND CONCLUSIONS: Based on results, mentioned mixture is safe and effective in improving life quality of patients suffering from BPH.

Consumption of echium oil increases EPA and DPA in blood fractions more efficiently compared to linseed oil in humans.

BACKGROUND: A plant-based strategy to improve long-chain (LC) omega (n)-3 PUFA supply in humans involves dietary supplementation with oils containing α-linolenic acid (ALA) alone or in combination with stearidonic acid (SDA). The study aimed to compare the effects of echium oil (EO) and linseed oil (LO) on LC n-3 PUFA accumulation in blood and on clinical markers. METHODS: In two double-blind, parallel-arm, randomized controlled studies, all volunteers started with 17 g/d run-in oil (2 weeks). Thereafter, subjects received diets enriched in study 1 with EO (5 g ALA + 2 g SDA; n = 59) or in study 2 with LO (5 g ALA; n = 9) daily for 8 weeks. The smaller control groups received fish oil (FO; n = 19) or olive oil (OO; n = 18). Participants were instructed to restrict their dietary n-3 PUFA intake throughout the studies (e.g., no fish). To investigate the influence of age and BMI on the conversion of ALA and SDA as well as clinical markers, the subjects recruited for EO and LO treatment were divided into three subgroups (two age groups 20-35 y; 49-69 y with BMI 18-25 kg/m(2) and one group with older, overweight subjects (age 49-69 y; BMI >25 kg/m(2)). RESULTS: In plasma, red blood cells (RBC), and peripheral blood mononuclear cells (PBMC), EPA and docosapentaenoic acid (DPA) were ~25 % higher following EO compared to LO. Comparing all treatments, the effectiveness of increasing EPA and DPA in plasma, RBC, and PBMC was on average 100:25:10:0 and 100:50:25:0 for FO:EO:LO:OO, respectively. EO led to a lower arachidonic acid/EPA-ratio compared to LO in plasma, RBC, and PBMC. Following EO, final DHA was not greater compared to LO. Higher BMI correlated negatively with increases in plasma EPA and DPA after EO supplementation, but not after LO supplementation. Decreasing effect on plasma LDL-C and serum insulin was greater with EO than with LO. CONCLUSIONS: Daily intake of SDA-containing EO is a better supplement than LO for increasing EPA and DPA in blood. However, neither EO nor LO maintained blood DHA status in the absence of fish/seafood consumption. TRIAL REGISTRATION: ClinicalTrials.gov Reg No. NCT01856179; ClinicalTrials.gov Reg No. NCT01317290.

Effect of ruminally unprotected Echium oil on milk yield, composition and fatty acid profile in mid-lactation goats.

This study investigated the effects on goat milk yield and composition of a diet supplemented with Echium plantagineum oil (EPO). Twenty-four mid-lactation multiparous Camosciata goats were divided into two balanced groups and fed for 44 d a diet based on hay and concentrate, supplemented (EPO group, Echium) or not (CON group, control) with 40 ml of ruminally unprotected EPO. Individual milk yield was recorded and individual milk samples were collected at 11, 22, 33, and 44 d after supplementation. Milk samples were analysed for milk components and fatty acids (FA). Data were statistically analysed by repeated-measures analysis of variance. Milk yield, protein and lactose contents were significantly higher in EPO than CON group. The inclusion of EPO significantly decreased total saturated FA and total branched-chain FA, and contemporarily sharply increased trans biohydrogenation intermediates (P ⩽ 0.001). Milk concentration of α-linolenic, stearidonic and γ-linolenic acids increased by 23, 1000 and 67%, respectively (P ⩽ 0.001). Due to extensive ruminal biohydrogenation, their apparent transfer rate was less than 3%. As a consequence, the milk concentrations of very long-chain (VLC) polyunsaturated fatty acids (PUFA), such as eicosapentaenoic (20:5 n-3) and dihomo-γ-linolenic (20:3 n-6) acids, significantly increased with EPO treatment, but values remained very low. Docosahexaenoic acid (22:6 n-3) was undetectable in all analysed milk samples. Results show that ruminally unprotected EPO can enhance milk yield and protein and improve the overall goat milk FA profile. However, this kind of supplementation cannot be considered a valuable strategy to develop goat functional dairy products enriched with VLC n-3 PUFA for human consumption.

Botanical oils enriched in n-6 and n-3 FADS2 products are equally effective in preventing atherosclerosis and fatty liver.

Echium oil (EO), which is enriched in 18:4 n-3, the immediate product of fatty acid desaturase 2 (FADS2) desaturation of 18:3 n-3, is as atheroprotective as fish oil (FO). The objective of this study was to determine whether botanical oils enriched in the FADS2 products 18:3 n-6 versus 18:4 n-3 are equally atheroprotective. LDL receptor KO mice were fed one of four atherogenic diets containing 0.2% cholesterol and 10% calories as palm oil (PO) plus 10% calories as: 1) PO; 2) borage oil (BO; 18:3 n-6 enriched); 3) EO (18:4 n-3 enriched); or 4) FO for 16 weeks. Mice fed BO, EO, and FO versus PO had significantly lower plasma total and VLDL cholesterol concentrations; hepatic neutral lipid content and inflammation, aortic CE content, aortic root intimal area and macrophage content; and peritoneal macrophage inflammation, CE content, and ex vivo chemotaxis. Atheromas lacked oxidized CEs despite abundant generation of macrophage 12/15 lipooxygenase-derived metabolites. We conclude that botanical oils enriched in 18:3 n-6 and 18:4 n-3 PUFAs beyond the rate-limiting FADS2 enzyme are equally effective in preventing atherosclerosis and hepatosteatosis compared with saturated/monounsaturated fat due to cellular enrichment of ≥20 PUFAs, reduced plasma VLDL, and attenuated macrophage inflammation.

The use of endemic Iranian plant, Echium amoenum, against the ethyl methanesulfonate and the recovery of mutagenic effects.

In this study, potential genotoxic effects of ethyl methanesulfonate (EMS) that caused mutagenicity in a variety of organisms were tried to resolve by the methanol and chloroform extract of Echium amoenum (EAmet and EAchl) Fisch. & C.A. Mey. from the family of Boraginaceae, which is an endemic plant, and is used as an alternative treatment among public in Iran. Somatic mutation and recombination test with Drosophila wing was used to determine the genotoxic and antigenotoxic effects in our investigations. For this purpose, 3-day-old transheterozygous larvae of mwh/flr(3) genotype of Drosophila melanogaster were used in all our experiments. The larvae were fed chronically on the Drosophila instant medium (DIM) including 1 ppm EMS. However, in another application group, different concentrations (1, 2 and 4 ppm) of EAmet and EAchl were added to DIM including 1 ppm EMS (EMS + EAmet and EMS + EAchl). Then, for the matured individuals, wing preparates were prepared within the mediums that include control group that has only DIM, negative control group that contains dimethyl sulfoxide and application groups in different concentrations that contain EMS, EMS + EAmet and EMS + EAchl. Clone induction frequency for the normal wing phenotype of EMS application group was observed to be 2.00. In the EMS + EAmet application group, the value of 1 ppm EAmet is 1.49, value of 2 ppm EAmet is 1.08 and value of 4 ppm EAmet is 0.72; in the EMS + EAchl application group, the value of 1 ppm is EAchl 1.33, value of 2 ppm EAchl is 0.67 and value of 4 ppm EAchl is 0.56 were determined. This decrease observed between EMS and all application groups in terms of total induction frequency is statistically significant (p < 0.05). These results concluded that chloroform extracts were more effective than the methanol extracts of E. amoenum.

Metabolic Profiling of Pyrrolizidine Alkaloids in Foliage of Two Echium spp. Invaders in Australia--A Case of Novel Weapons?

Metabolic profiling allows for simultaneous and rapid annotation of biochemically similar organismal metabolites. An effective platform for profiling of toxic pyrrolizidine alkaloids (PAs) and their N-oxides (PANOs) was developed using ultra high pressure liquid chromatography quadrupole time-of-flight (UHPLC-QTOF) mass spectrometry. Field-collected populations of invasive Australian weeds, Echium plantagineum and E. vulgare were raised under controlled glasshouse conditions and surveyed for the presence of related PAs and PANOs in leaf tissues at various growth stages. Echium plantagineum possessed numerous related and abundant PANOs (>17) by seven days following seed germination, and these were also observed in rosette and flowering growth stages. In contrast, the less invasive E. vulgare accumulated significantly lower levels of most PANOs under identical glasshouse conditions. Several previously unreported PAs were also found at trace levels. Field-grown populations of both species were also evaluated for PA production and highly toxic echimidine N-oxide was amongst the most abundant PANOs in foliage of both species. PAs in field and glasshouse plants were more abundant in the more widely invasive species, E. plantagineum, and may provide competitive advantage by increasing the plant's capacity to deter natural enemies in its invaded range through production of novel weapons.

Dietary echium oil increases long-chain n-3 PUFAs, including docosapentaenoic acid, in blood fractions and alters biochemical markers for cardiovascular disease independently of age, sex, and metabolic syndrome.

Dietary supplementation with echium oil (EO) containing stearidonic acid (SDA) is a plant-based strategy to improve long-chain (LC) n-3 (ω-3) polyunsaturated fatty acid (PUFA) status in humans. We investigated the effect of EO on LC n-3 PUFA accumulation in blood and biochemical markers with respect to age, sex, and metabolic syndrome. This double-blind, parallel-arm, randomized controlled study started with a 2-wk run-in period, during which participants (n = 80) were administered 17 g/d run-in oil. Normal-weight individuals from 2 age groups (20-35 and 49-69 y) were allotted to EO or fish oil (FO; control) groups. During the 8-wk intervention, participants were administered either 17 g/d EO (2 g SDA; n = 59) or FO [1.9 g eicosapentaenoic acid (EPA); n = 19]. Overweight individuals with metabolic syndrome (n = 19) were recruited for EO treatment only. During the 10-wk study, the participants followed a dietary n-3 PUFA restriction, e.g., no fish. After the 8-wk EO treatment, increases in the LC n-3 metabolites EPA (168% and 79%) and docosapentaenoic acid [DPA (68% and 39%)] were observed, whereas docosahexaenoic acid (DHA) decreased (-5% and -23%) in plasma and peripheral blood mononuclear cells, respectively. Compared with FO, the efficacy of EO to increase EPA and DPA in blood was significantly lower (∼25% and ∼50%, respectively). A higher body mass index (BMI) was associated with lower relative and net increases in EPA and DPA. Compared with baseline, EO significantly reduced serum cholesterol, LDL cholesterol, oxidized LDL, and triglyceride (TG), but also HDL cholesterol, regardless of age and BMI. In the FO group, only TG decreased. Overall, daily intake of 15-20 g EO increased EPA and DPA in blood but had no influence on DHA. EO lowered cardiovascular risk markers, e.g., serum TG, which is particularly relevant for individuals with metabolic syndrome. Natural EO could be a noteworthy source of n-3 PUFA in human nutrition.

Bioavailability and potential uses of vegetarian sources of omega-3 fatty acids: a review of the literature.

Presently alpha-linolenic acid (ALA) is the most widely used vegetarian LC3PUFA, but only marginal amounts are converted into eicosapentaenoic (EPA) and docosahexaenoic acid (DHA); both of which are strongly related to human health. Currently, fish oils represent the most prominent dietary sources of EPA and DHA; however, these are unsuitable for vegetarians. Alternative sources include flaxseed, echium, walnut, and algal oil but their conversion to EPA and DHA must be considered. The present systematic review sets out to collate information from intervention studies examining the bioavailability of alternative vegetarian long chain omega-3 (n-3) polyunsaturated fatty acids (LC3PUFA) sources. Ten key papers published over the last 10 years were identified with seven intervention studies reporting that ALA from nut and seed oils was not converted to DHA at all. Three studies showed that ingestion of micro-algae oil led to significant increases in blood erythrocyte and plasma DHA. Further work is now needed to identify optimal doses of alternative vegetarian LC3PUFAs and how these can be integrated within daily diets. The potential role of algal oils appears to be particularly promising and an area in which further research is warranted.