Effects of fungal endophytes and arbuscular mycorrhizal fungi on growth of Echium vulgare and alkannin/shikonin and their derivatives production in roots.

Endophytic fungi as well as arbuscular mycorrhizal fungi (AMF) are known to stimulate plant growth and production of secondary metabolites in medicinal plants. Here, 10 endophytic fungi isolated from roots of wild Alkanna tinctoria plants and 5 AMF purchased from the Glomeromycota in vitro collection were evaluated, during two successive three-month greenhouse experiments, on the growth of Echium vulgare and alkannin/shikonin and their derivatives (A/Sd) production in the roots. Some of the endophytic fungi tested significantly increased plant growth parameters as compared to the control: Cladosporium allicinum, Cadophora sp., Clonostachys sp., Trichoderma hispanicum and Leptosphaeria ladina increased root volume, Plectosphaerella sp. And T. hispanicum root fresh weight and root water retention and T. hispanicum plant water retention. However, none of these fungi impacted A/Sd production. Conversely, none of the AMF strains tested impacted plant growth parameters, but those inoculated with Rhizophagus intraradices MUCL 49410 had a significantly higher concentration of alkannin/shikonin (A/S), acetyl-A/S, ß,ß- dimethylacryl-A/S, isovaleryl-A/S and total A/Sd, compared to the control plants. Further studies are needed to investigate the mechanisms involved in the production of A/Sd in plants associated to specific endophytic fungi/AMF and on the cultivation conditions required for optimal production of these compounds.

Effects of sowing date on photosynthetic characteristics, chlorophyll fluorescence and yield of different Echium plantagineum L. cultivars.

The seed oil of Echium plantagineum L. is rich in unsaturated fatty acids. With the gradual development of the value of echium oil in food, medical care and cosmetics, the corresponding market demand has also increased. The selection of suitable cultivars and the increase of yield per unit area has also become one of the main objectives of current breeding and cultivation of E. plantagineum. To effectively use the local photothermal resources, to improve the use of light energy by E. plantagineum, and to enhance the growth and yield of E. plantagineum. E. plantagineum cultivars Blue Bedder and Mixed Bedding were used as research subjects to study the effects of different sowing dates (1 May, 8 May, 15 May, 22 May and 29 May) on the photosynthetic characteristics and yield of E. plantagineum. Under the same cultivar conditions, with the delay in sowing date, the leaf chlorophyll content (SPAD), photosynthetic rate (Pn), transpiration rate (Tr), stomatal limitation value (Ls), photochemical quenching (qP), electron transfer rate (ETR), actual photochemical efficiency (ΦpsII) and yield of Blue Bedder decreased and reached a maximum at T1, while the SPAD, Pn, Tr, water use efficiency (WUE), Ls, initial fluorescence (Fo), maximum fluorescence (Fm), qP, ETR, ΦpsII and yield of Mixed Bedding reached the maximum at T4. Blue Bedder should be sown early at T1 and Mixed Bedding late at T4 during planting, which will help to improve the photosynthetic characteristics and grain yield of E. plantagineum.

The effect of infusion time on Echium amoenum extract -induced hepatotoxicity in vitro.

Echium amoenum is an annual herb native to the northern mountains of Iran which has medicinal application. Petals of Echium amoenum (Gole-Gavzaban) is one of the most valuable medicinal plants in Iranian folk medicine. The dry petals of E. amoenum have long been used as a sedative, tonic, anxiolytic and as a treatment for sore throat, cough and inflammation. Previous studies have shown that petals of E. amoenum contain four toxic pyrrolizidine alkaloids but conflicting results have been acquired in experimental studies investigating the hepatotoxicy of E. amoenum. However, the direct effect of E. amoenum on liver cells and the complete mechanisms of its possible cytotoxic effects toward these cells remain to be defined. The main aim of this study was to assay the mechanisms underlying the toxic effects of E. amoenum toward hepG2 cells. E. amoenum extract was obtained by infusion of dried petals in hot water (90 centigrade) for 15 or 30 min. Cell viability and mechanistic parameters were determined following 12 h incubation of hepG2 with E. amoenum extract that was obtained after 15 or 30 min infusion. The results indicated that E. amoenum extract exerts cytotoxic effects on hepG2 cells, probably through mitochondrial and lysosomal damage induced by glutathione depletion and oxidative stress.

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.

Design and in vitro effectiveness evaluation of Echium amoenum extract loaded in bioadhesive phospholipid vesicles tailored for mucosal delivery.

The Echium amoenum Fisch. and C.A. Mey. (E. amoenum) is an herb native from Iranian shrub, and its blue-violet flowers are traditionally used as medical plants. In the present study, an antioxidant phytocomplex was extracted from the flowers of E. amoenum by ultrasounds-assisted hydroalcoholic maceration. The main components, contained in the extract, have been detected using HPLC-DAD, and rosmarinic acid was found to be the most abundant. The antioxidant power of the extract along with the phenolic content were measured using colorimetric assays. The extract was loaded in liposomes, which were enriched adding different bioadhesive polymers (i.e., mucin, xanthan gum and carboxymethyl cellulose sodium salt) individually or in combination. The main physico-chemical properties (i.e. size, size distribution, surface charge) of the prepared vesicles were measured as well as their stability on storage. The viscosity of dispersion and the ability of vesicles to interact with mucus were evaluated measuring their stability in a mucin dispersion and mobility in a mucin film. The biocompatibility and the ability of the formulations to protect keratinocytes from damages caused by hydrogen peroxide and to promote the cell migration were measured in vitro.

Olive and echium oil gelled emulsions: Simulated effect of processing temperature, gelling agent and in vitro gastrointestinal digestion on oxidation and bioactive compounds.

The impact and relative relevance of in vitro gastrointestinal digestion, processing temperature (room temperature or 180 °C), and gelling agent (GA) (carrageenan and alginate) on the bioactive compounds and oxidation status of olive and echium oils gelled formulations with 40% lipid incorporation was assessed. In vitro digestion was not affected by the GA, with >90% lipolysis in all formulations, but was the most relevant variable, promoting oxidation (MDA) regardless of the oil type, GA or temperature applied. Tocopherols and phenolic decreased with digestion, which could be interpreted as a protective response to pro-oxidative conditions during digestion. Temperature decreased olive oil phenolics. Gelification of echium oil using alginate reduced secondary oxidation products formation in comparison with carrageenan, with oxidation degrees after digestion equivalent to those shown with olive oil. The use of alginate with olive oil resulted in the most stable formulations, although not protecting its minor bioactive compounds from thermal degradation.

Secondary metabolite contents and safety assessment study of the aqueous extract from the Algerian Echium trygorrhizum Pomel roots.

ETHNOPHARMACOLOGICAL RELEVANCE: Species of Echium trygorrhizum Pomel, belonging to the Boraginaceae family, grow wild in North Africa. This plant is used in traditional Algerian medicine for the treatment of Diabetes, Jaundice and Tonsillitis. AIM OF THE STUDY: To our knowledge, no work has been done on the phytochemistry and biological activities of this plant. Moreover, no study has yet corroborated that the use of this plant is safe. Therefore, the present study was carried out to gather information on the various medicinal uses of this plant and to evaluate the total phenolic content and assess its safety after acute and sub-acute toxicity tests with rodents (mice and rats). MATERIAL AND METHODS: An ethnopharmacological survey was carried out using a direct questionnaire. It included some questions relating to the plant. The content of polyphénols, flavonoids and tannins present in the aqueous extract was determined by colorimetric methods. In the acute toxicity tests, three groups each containing five males and five females Albino mice were formed. The control group received water, while the test groups received different doses of aqueous extract (2000 and 5000) mg/kg body weight. In the subacute toxicity study, different doses (250, 500 and 1000 mg/kg) of the aqueous extract were administered to male and female rats for 28 consecutive days. Hematological analysis, biochemical parameters estimation and histopathological examination have been determined at the end of the treatment. RESULTS: Data from the ethnopharmacological survey showed that 25, 31% of people used this plant in Algeria as a traditional medicine for the treatment of Jaundice (100%), Diabetes (28, 33%) and Tonsillitis (10%). Results also revealed that aqueous extract contains high amounts of polyphenols, flavonoids and tannins. The extract did not cause any toxicity during the 14-days observation period after a single dose of 5000 mg/kg was administered to mice, with LD50 values greater than 5000 mg/kg. In a further step, the administration of the aqueous extract at all dose levels (250, 500, and 1000 mg/kg) to male and female rats during the 28-days study did not result in any deaths or behavioral changes, and there were no changes in body weight, relative organ weights, or food consumption as a result of this experiment. However, male rats showed a significant difference in relative liver weight in the high dose group (1000 mg/kg) (p < 0.05) and the satellite group (p < 0.01). In female rats, liver weight significantly increased only in the satellite group compared with the control group (p < 0.01). The results showed an increase in red blood cells (RBC), hemoglobin (Hb), and hematocrit (Hct) in the female groups. However, a clear decrease was observed in plasma ALP enzyme activities in females treated with the doses of 250 and 1000 mg/kg (p < 0.05) after the treatment periods. CONCLUSION: The study indicated that the single dose of 5 g/kg AQE can be considered relatively safe as it did not cause death or any signs of toxicity in mice. Repeated oral administration of AQE at doses below 250 g/kg/day for 28 consecutive days can be considered relatively safe.

Echium amoenum L. Ethanol Extract Protects Retinal Ganglion Cell after Glutamate and Optic Nerve Crush Injury.

The development of low-cost and effective natural products for treating neuron degenerative diseases have proven to be safe and potentially effective. Echium amoenum L. (Boraginaceae) is an annual herb that grows wildly in Europe and western Asia. The aim of this study was to evaluate the neuroprotective properties of an ethanol extract of E. amoenum L. The effects of E. amoenum L. extract on oxidative stress were measured in the rat R28 retinal precursor cell line. Furthermore, the protective role of the extract on the glutamate-induced and optic nerve crush (ONC) injury-induced cell death were evaluated in vitro and in vivo, respectively. Our results showed that the ethanol extract of E. amoenum L. prevented the glutamate-induced decrease in cell viability and increase in cell death in R28 cells and suppressed the overproduction of ROS induced by glutamate. Moreover, the extract significantly inhibited microglial activation and optic nerve damage induced by ONC injury in mice. In addition, the mechanism was attributed to the ability of the extract to decrease NF-κB pathway activation and its downstream inflammatory cytokine production. In conclusion, E. amoenum L. ethanol extract had a potent neuroprotective effect against glutamate-induced and ONC-induced cell death. This is likely due to its antioxidant and anti-inflammatory properties.

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).

Comparing the Effects of a Herbal Drug based on Echium Amoenum With Fluvoxamine in the Treatment of Adolescents with Obsessive-compulsive Disorder.

BACKGROUND: Obsessive-compulsive disorder (OCD) is a severe and debilitating neuropsychiatric condition. Although selective serotonin reuptake inhibitors, tricyclic antidepressants, and cognitive- behavioral therapy are the first-line medication and treatment for OCD, an estimated 30% of patients are treatment-resistant, and complete functional recovery is rare. Natural products as adjuvant or alternative therapies should be examined to find safer and more effective ways to manage OCD. OBJECTIVES: To investigate the potential benefits of a combined herbal drug based on Echium amoenum in treating OCD. METHODS: Design and Setting: In the psychiatric clinics of Mashhad University of Medical Sciences, 40 patients who met the criteria for the obsessive-compulsive disorder based on DSM-5 were studied in a parallel, double-blind, randomized clinical trial. INTERVENTION: Subjects were randomly assigned to receive Echium amoenum-Melissa officinalis syrup and fluvoxamine or placebo syrup and fluvoxamine for 8 weeks. OUTCOME MEASURES: The efficacy of treatment and recurrence of disease were surveyed and compared according to the Yale-Brown Obsessive Compulsive Scale at weeks 0, 4, and 8. RESULTS: Evaluation at the 4th and 8th week showed no significant differences between the two groups (p-value = 0.11, p-value = 0.445, respectively). At the 8th week of treatment, patients in the intervention group showed a remarkable reduction in scores on the Yale-Brown Obsessive-Compulsive Scale questionnaire (p- value= 0.003), and patients in the control group didn't ((p- value= 0.180). This study showed that the E.amoneum-M.officinalis syrup was not significantly more efficacious than the fluvoxamine tablet, but the intervention group showed a significant improving trend (p-value= 0.001). CONCLUSION: While monotherapy is usually the gold standard methodology, combination or augmentation therapy may also be of merit. Consequently, studies with larger sample sizes and the inclusion of para-clinical assessments such as serologic tests can further shed light on the mechanism of action of the E. amoneum- M. officinalis syrup and deepen our understanding of its effects.

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.

Fatty acid profile, sterols, and squalene content comparison between two conventional (olive oil and linseed oil) and three non-conventional vegetable oils (echium oil, hempseed oil, and moringa oil).

New sources of bioactive compounds are constantly explored for reformulating healthier foods. This work aimed to explore and characterize the fatty acid profile and sterol content of three non-conventional oils used in functional food products (hempseed oil, moringa oil, and echium oil) and to compare them with two conventional ones (extra virgin olive oil [EVOO] and linseed oil). Oxidative stability was assessed by determining their acidity value and peroxide content. All oils showed adequate values for acidity and oxidation status. Echium and hempseed oils showed a high content of polyunsaturated fatty acids (>70%), especially omega-3 fatty acids, while moringa oil was rich in oleic acid. Echium oil, hempseed oil, and moringa oil presented higher sterol content than EVOO, but lower than that of linseed oil. Sitosterol was the most abundant sterol in all samples (97.88-275.36 mg/100 g oil), except in echium oil, where campesterol (170.62 mg/100 g oil) was the major sterol. Squalene was only found in significant amounts in EVOO. In conclusion, non-conventional oils seem to be interesting sources of bioactive compounds and have great potential for the food industry. PRACTICAL APPLICATION: Non-conventional vegetable oils can be used as alternative sources of lipids in a variety of food products. Additionally, these oils have great potential to be included in the formulation of functional ingredients for the delivery of omega-3 fatty acids, antioxidants, fiber, among others.

An exploratory method to determine the plant characteristics affecting the final yield of Echium amoenum Fisch. & C.A. Mey. under fertilizers application and plant densities.

Employing of advanced statistical methods to quantify agricultural information has helped to carry out targeted planning to alleviate the problems of farmers, researchers and policy section. One of these exploratory methods, is multivariate statistical analysis that examines and models the relationship between variables. Considering the importance of Echium amoenum and its use growing trend in traditional medicine and the pharmaceutical industry, also the lack of information on the correlations between its yield and morpho physiological traits, the objective of this study was to determine the causality path in which the Echium amoenum characteristics affects the yield of Echium amoenum as regards of application of organic and chemical fertilizers under different plant densities. The employed method revealed that organic fertilizers increased flower yield compared with the control. The flower yield as a result of application of compost, vermicompost and cattle manure were increased by 25, 28, and 27% compared with the control, respectively. The results of multiple regression showed that variables of plant height, shoot dry weight, flower number per plant were the main factors affected the flower yield. The relative contribution of shoot dry weight was 16 and 25% more than plant height and flower number per plant, respectively. Causality analysis identified that shoot dry weight per plant had indirect effect on flower yield in different paths, as mainly was imposed through plant height considering the path coefficients. This study suggests that optimum production of Echium amoenum with application of ecological inputs along with effective agronomical managements of the causal paths of flower yield forming, including increase in shoot yield and plant height could be achieved through an ecological cropping system with reduced costs and no health concerning due to agrochemicals residual.

Comparative analysis of the main medicinal substances and applications of Echium vulgare L. and Echium plantagineum L.: A review.

ETHNOPHARMACOLOGICAL RELEVANCE: Echium vulgare L. and Echium plantagineum L. originated in the Mediterranean, and were later domesticated in Africa, America, Asia, Europe and Oceania, where they were widely used to treat many diseases including cough, urinary tract infection, fever, inflammation and muscle strain. AIM OF THE STUDY: The purpose of this review is to provide scientific literature on the traditional uses, bioactive chemical components and pharmacological activities of two species of Echium, and to critically analyze the information provided, so as to understand the current work on these two species and explore the possible prospect of this plant in pharmaceutical research. METHODS: Systematic review and meta-analysis were conducted according to Prisma guidelines, and the related literatures searched on Google Academic, Science Direct, Baidu Scholars and China National Knowledge Infrastructure (CNKI) up to June 2021 were reviewed. The key words used are: Echium, E.vulgare, E.plantagineum, plant components, chemical components, pharmacological activities, pharmaceutical products and applications. Thereafter all eligible studies are analyzed and summarized in this review. The selection of manuscripts is based on the following inclusion criteria: the article has years of research or publication, is published in English, Portuguese or Spanish and Chinese, and there are keywords in the title, abstract, keywords or full text of the article. For the selection of manuscripts, first, select articles according to titles, then summarize them, and finally, analyze the full text of the publication. Elimination criteria: 1. Duplicate reports; 2. There are research design defects and poor quality; 3. Incomplete data and unclear ending effect; 4. The statistical method is wrong and cannot be corrected. RESULTS: The pharmacological characteristics of E.vulgare and E.plantagineum can basically support their traditional use, but the medicinal substances contained in them are quite different in composition and content, and the development and application of corresponding products are also different. CONCLUSIONS: At present, there is little clinical data about drugs related to the two species, and more research is needed in the future, especially human experiments and clinical trials, to evaluate the cellular and molecular mechanisms based on pharmacological, biological activity and safety studies, and to provide more powerful scientific basis for their traditional medicinal properties. In addition, the further application and development of the medicinal products of E.vulgare and E.plantagineum still need to be precise and identified, so as to give full play to their medicinal potential.

Chemical characterization of Echium plantagineum seed oil obtained by three methods of extraction.

Echium seed oil has been considered an important alternative source of omega 3 fatty acids (n-3 FA) for human consumption. Considering the oxidative instability of n-3 FA richer oils, the objective of this study was to determine the chemical and sensory parameters of the oil obtained from Echium plantagineum seeds obtained by three extraction methods (hydraulic press: HYD; continuous screw press: PRESS; and solvent technique: SOLV). Stearidonic acid (C18:4, n3), the most important n-3 FA present in the oil, changed from 12.5% to 12.7%. Regarding the minor compounds, PRESS sample showed the highest concentration of gamma-tocopherol (782.24 mg/kg oil), while SOLV samples presented the highest amount of ß-sitosterol (73.46 mg/100 g) with no difference of campesterol concentration (159.56 mg/100 g) among the samples. Higher values of total phenolics (19.65 mg GAE/kg oil) and ß-carotene (34.83 mg/kg oil) were also found in the SOLV samples, suggesting the influence of hexane in the extraction of these bioactive compounds. High resolution mass spectrometry identified caffeic acid and its derivatives as the main phenolic compounds present in the echium oil. PRESS sample showed the best oxidative stability as measured by PV (0.61 mmol/kg oil) and malondialdehyde (173.13 µmol), probably due to faster time of processing compared to HYD and SOLV samples. Our data showed that the extraction method changed the chemical composition of the minor compounds in the echium oil, but these alterations did not reduce its nutritional quality or sensory acceptability. PRACTICAL APPLICATION: Echium oil represents a great potential source of omega 3 fatty acids, but there is not enough information about its oxidative stability and chemical composition, especially toward minor compounds. Our study characterizes echium oil composition obtained from three extraction methods, contributing to amplify the technical information about this important alternative oil for human consumption.

Light quality and dormancy overcoming in seed germination of Echium plantagineum L. (Boraginaceae) / Qualidade da luz e superação de dormência na germinação de sementes de Echium plantagineum L. (Boraginaceae)

Abstract Light is considered a factor that influences the seed germination of many weed species, and it can signal whether the environmental conditions are favorable or are not favorable for germination. We aimed to study if there is an influence of light quality and dormancy overcoming in seed germination of Echium plantagineum L. We carried out a 2 x 6 factorial experiment, with and without dormancy overcoming with potassium nitrate followed by immersion in gibberellic acid; six light qualities, obtained through the light filters: blue, green, red, far-red, white light and absence of light. The evaluations performed were germination speed index (GSI), average germination time (AGT), germination at the four and 14 days after seeding (DAS), accumulated germination and relative frequency of germination. We observed significant interaction among the light qualities and seed dormancy overcoming or not for the studied variables. There was no significant effect of light qualities, in the evaluated variables, when performing dormancy overcoming, presenting germination above 90% in all the light qualities. However, without dormancy overcoming, we observed greater GSI, germination at four and 14 DAS for the red light filter with 5, 4, 29 and 45%, respectively. When the seeds were submitted to the absence of light, and without dormancy overcoming, there was only 7% of germination at 14 DAS. The seeds of E. plantagineum presented greater germination under incidence of red light, without dormancy overcoming, being classified as preferably positively photoblastics, provided that the dormancy is not overcome.

Resumo A luz é considerada um fator que influencia a germinação das sementes de muitas espécies de plantas daninhas, podendo sinalizar se as condições ambientais são favoráveis ou não para a germinação. Objetivou-se estudar se há influência da qualidade da luz e superação de dormência na germinação de sementes de Echium plantagineum L. Realizou-se um experimento fatorial 2 x 6, com e sem superação de dormência com nitrato de potássio seguido pela imersão em ácido giberélico; seis qualidades de luz, obtidas através de filtros de luz: azul, verde, vermelho, vermelho-distante, luz branca e ausência de luz. As avaliações realizadas foram índice de velocidade de germinação (IVG), tempo médio de germinação (TMG), germinação aos quatro e 14 dias após a semeadura (DAS), germinação acumulada e frequência relativa de germinação. Observou-se interação significativa entre as qualidades de luz e a superação ou não de dormência das sementes para as variáveis estudadas. Não houve efeito significativo das qualidades de luz, nas variáveis avaliadas, ao realizar superação de dormência, apresentando germinação acima de 90% em todas as qualidades de luz. Todavia, sem superação de dormência, observou-se maior IVG, germinação aos quatro e 14 DAS para o filtro de luz vermelha com 5,4, 29 e 45%, respectivamente. Quando as sementes foram submetidas à ausência de luz, e sem superação de dormência, houve apenas 7% de germinação aos 14 DAS. As sementes de E. plantagineum apresentam maior germinação sob incidência de luz vermelha, sem superação de dormência, sendo classificadas como fotoblásticas positivas preferenciais, desde que não seja superada a dormência.

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.

Pseudotargeted Lipidomics Strategy Enabling Comprehensive Profiling and Precise Lipid Structural Elucidation of Polyunsaturated Lipid-Rich Echium Oil.

Echium oil has great nutritional value as a result of its high content of α-linolenic acid (ALA, 18:3ω-3) and stearidonic acid (SDA, 18:4ω-3). However, the comprehensive lipid profiling and exact structural characterization of bioactive polyunsaturated lipids in echium oil have not yet been obtained. In this study, we developed a novel pseudotargeted lipidomics strategy for comprehensive profiling and lipid structural elucidation of polyunsaturated lipid-rich echium oil. Our approach integrated untargeted lipidomics analysis with a targeted lipidomics strategy based on Paternò-Büchi (PB)-tandem mass spectrometry (MS/MS) using 2-acetylpyridine (2-AP) as the reaction reagent, allowing for high-coverage lipid profiling and simultaneous determination of C═C locations in triacylglycerols (TGs), diacylglycerols (DGs), free fatty acids (FFAs), and sterol esters (SEs) in echium oil. A total of 209 lipid species were profiled, among which 162 unsaturated lipids were identified with C═C location assignment and 42 groups of ω-3 and ω-6 C═C location isomers were discovered. In addition, relative isomer ratios of certain groups of lipid C═C location isomers were revealed. This pseudotargeted lipidomics strategy described in this study is expected to provide new insight into structural characterization of distinctive bioactive lipids in food.

Combination of natural strategies to improve the oxidative stability of echium seed oil.

Echium seed oil is an alternative source of omega 3 fatty acids but it is highly susceptible to oxidation. A combination of three natural strategies was proposed in this study aiming to improve the oxidative stability of echium oil obtained by pressing (PO) or solvent extraction (PSO), kept in the storage condition for 180 days or during the consumption for 30 days. Our results showed that the reduction of temperature was sufficient to keep the oil stable during storage for both samples. During the consumption time, the best stability was achieved by adding a mixture of antioxidants, composed of sinapic (500 ppm), ascorbic (250 ppm), and citric (150 ppm) acids, and/or 20% of high oleic sunflower oil. The combined strategies promoted a 34 to 80% reduction of peroxide value and 0 to 85% reduction of malondialdehyde concentrations in the samples, showing to be a feasible and natural alternative to improve the oxidative stability of echium oil. PRACTICAL APPLICATION: Our study successfully applied an optimized combination of simple and low-cost strategies to enhance the chemical stability of echium seed oil. As the use of echium oil expands around the world, the oil industry and final consumers may benefit from our results to increase the oil shelf-life.

Hydromethanolic root and aerial part extracts from Echium arenarium Guss suppress proliferation and induce apoptosis of multiple myeloma cells through mitochondrial pathway.

Echium arenarium Guss is a Mediterranean plant traditionally used in healing skin wound and it was reported exhibiting potent antioxidant, antibacterial, and antiparasitic activities. However, antitumoral activities of this plant have not yet been explored. Here we investigated for the first time, root (EARE) and aerial part (EAAPE) extracts of E. arenarium Guss to examine cytotoxicity and apoptosis activation pathway on U266 human multiple myeloma (MM) cell line. We demonstrated that EARE and EAAPE decreased U266 cell viability in a dose dependent manner. Based on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, EARE was significantly two times more efficient (IC50 value 41 µg/ml) than EAAPE (IC50 value 82 µg/ml) considering 48 h of treatment. Furthermore, after 24 h of exposure to 100 µg/ml of EARE or EAAPE, cell cycle showed remarkable increase in sub-G1 population and a decrease of U266 cells proportion in G1 phase. In addition, EARE increased cell percentage in S phase. Moreover, analysis revealed that EAAPE or EARE induced apoptosis of U266 cells after 24 h of treatment. Interestingly, depolarization of mitochondrial membrane potential and activation of caspase 3/7 were demonstrated in treated U266 cells. Phytochemical analysis of E. arenarium extracts showed that EARE exhibited the highest content of total phenolic content. Interestingly, six phenolic compounds were identified. Myricitrin was the major compound in EARE, followed by luteolin 7-O-glucoside, resorcinol, polydatin, Trans-hydroxycinnamic acid, and hyperoside. These findings proved that an intrinsic mitochondria-mediated apoptosis pathway probably mediated the apoptotic effects of E. arenarium Guss extracts on U266 cells, and this will suggest several action plans to treat MM.