Glycoside Rutin Research Articles

QUANTITATIVE ESTIMATION OF RUTIN IN RUE (RUTA GRAVEOLENS L.) CULTIVATED IN IRAQ WITH THE EVALUATION OF ITS ANTIOXIDANT ACTIVITY

Objective: This study was designed to estimate the quantity of the flavonoid glycoside (rutin) as an important active compound in rue plant grown inIraq with the evaluation of its antioxidant activity.Methods: The cultivated rue plant at the garden of medicinal plant in the pharmacognosy and medicinal plants department was collected and dried at room temperature. Total flavonoids content was determined using aluminum chloride method, and a standard calibration curve is used for estimation of rutin glycoside. Antioxidant activity of the ethanolic extract was measured using the (1,1-diphenyl-2-picrylhydrazyl radical).Results: The results were showed that 100 g of the plant produced 7 g powdered extract, and each gram of the extract contains 133 mg rutin. The extract showed a positive antioxidant activity as compared with standard quercetin.Conclusion: Ruta graveolens plant contains the flavonoid glycoside rutin which showed antioxidant activity; therefore, further studies are required to identify other constituents in this plant.Keywords: Rue, Rutin, Flavonoids and antioxidant.

Growth, hydrolases and ultrastructure of Fusarium oxysporum as affected by phenolic rich extracts from several xerophytic plants

Fusarium oxysporum, the causal agent of rot and wilt diseases, is one of the most detrimental phytopathogens for the productivity of many economic crops. The present study was conducted to evaluate the potentiality of some xerophytic plants as eco-friendly approach for management of F. oxysporum. Phenolic rich extracts from five plants namely: Horwoodia dicksoniae, Citrullus colocynthis, Gypsophila capillaris, Pulicaria incisa and Rhanterium epapposum were examined in vitro. The different extracts showed high variability in their phenolic and flavonoid contents as well as total antioxidant capacity. A strong positive correlation existed between the antifungal activity of the tested extracts and their contents of both total phenolics and flavonoids (r values are 0.91 and 0.82, respectively). Extract of P. incisa was the most effective in reducing the mycelial growth (IC50=0.92mg/ml) and inhibiting the activities of CMCase, pectinase, amylase and protease by 36, 42, 58 and 55%, respectively. The high performance liquid chromatography analysis of P. incisa extract revealed the presence of eight phenolic acids along with five polyphenolic compounds. The flavonol, quercetin and its glycosides rutin and quercetrin were the most abundant followed by the phenolic acids, t-cinnamic, caffeic, ferulic and vanillic. P. incisa extract not only affects the growth and hydrolases of F. oxysporum but also induces ultrastructure changes in the mycelium, as revealed by transmission electron microscopy. To our knowledge, this is the first study to investigate the mechanisms underlying the antifungal activity of P. incisa.

Echinophora tenuifolia L. inflorescences: phytochemistry and in vitro antioxidant and anti-inflammatory properties in LPS-stimulated RAW 264.7 macrophages

During the last decades, different natural compounds have been demonstrated to modulate inflammatory pathways. In this study, methanolic extract of Echinophora tenuifolia L. inflorescences was investigated for its chemical composition and in vitro antioxidant and anti-inflammatory properties. Phytochemical profile was investigated by means of high-performance thin layer chromatography (HPTLC) and gas chromatography–mass spectrometry analyses. Myristic acid and palmitic acid were found to be the major constituents. Six terpenes were identified: α-phellandrene, o-cymene, dihydroactinidiolide, neophytadiene, phytol and β-amyrin, and two phenolic compounds: carvacrol and ferulic acid. HPTLC analysis of the ethyl acetate fraction highlighted the presence of the flavonoid glycosides rutin and quercitrin. This sample showed the best diphenylpicrylhydrazyl scavenging capacity, with an IC50 value equal to 40.39 μg/ml, and the strongest capacity to protect linoleic acid from peroxidation, as assessed by the β-carotene bleaching test (IC50 = 16.31 μg/ml). All samples inhibited nitric oxide production in cell supernatants in a dose-dependent manner, being the two apolar fractions (n-hexane and dichloromethane) even more active than the positive control indomethacin. A relevant biological activity was observed for dichloromethane fraction (IC50 value equal to 39.97 μg/ml). Obtained results indicate that this sample could be an excellent candidate for further investigations aimed at the development of new antioxidant and anti-inflammatory drugs.

Flow Cytometric Method for the Detection of Flavonoids in Cell Lines

Here, we describe an easy-to-use flow cytometric method using diphenylboric acid 2-amino ethyl ester (DPBA) stain for the detection of flavonoids in cells from human/animal origin. Flavonoid bioavailability and bioactivity depend on structure, conjugation and the cell type to which they are presented. We have studied cellular uptake of five flavonoids with different structures and conjugation forms. First, parameters including fixation method, technical and batch variability, and concentration were optimized. Second, uptake of two aglycones—quercetin and hesperetin—and their corresponding glycosides—rutin and hesperidin—in Caco-2 cells was compared. Third, the aglycone quercetin, glycoside rutin, and glucuronide baicalin were added to the Caco-2, HepG2, and CHO-K1 cell lines at 1, 10, and 20 µM concentrations and cellular uptake was measured after 1, 4, and 7 h. We conclude that quercetin was taken up by cells in a dose-dependent way, and that HepG2 cells had the highest uptake factors, followed by CHO-K1 and Caco-2 cells. Confocal microscopy showed cell type–dependent localization of quercetin in the cell membrane and cytoplasm. No uptake of flavonoid glycosides was detected. This flow cytometric method can be used for future research unravelling mechanisms behind flavonoid bioactivity in health and disease at the cellular level.

Photoinduced oxidation reactions of the pendent macrocyclic complexes: Al(III) phthalocyanine and Ni(II) annulene mechanistic considerations

The goal of the work was to expand the understanding of the photoinduced redox reactions of the Al(III) sulfonated phthalocyanine, HOAlIIItspc, and Ni(II) tertramethyldibenzotetraazaannnulene, [NiII(tmdbTAA)], as components of polymeric structures in which poly(ethyleneimino) and poly(isobutyl-alt-maleate) are the respective backbones of poly(HOAlIIItspc) and poly([NiII(tmdbTAA)]). Saccharide and flavonoid glycosides form adducts that quench the photo-generated oxidizing radicals in the case of poly(HOAlIIItspc) with rate constants≈1×104s−1, largely independent of reductant structure. The charge-separated intermediate, CS, in the case of poly([NiII(tmdbTAA)]) shows greater reactivity with the saccharides and flavonoid glycosides, particularly with the flavonoid glycoside rutin where the rate constant is 2.7×106s−1. While phthalocyanine-centered radicals in poly(HOAlIIItspc) were reduced by dAMP (adenosine-5-monophosphate) and CT-DNA, energy transfer with these reagents competed with the formation of the CS intermediate in poly([NiII(tmdbTAA)]). A study of the intensity of the upper conversion luminescence (λem<500nm), modulated by the concentration of dAMP suggests that this is attributed to the formation of adducts of poly([NiII(tmdbTAA)]) with dAMP that affect the deactivation of the electronically excited Ni(II) tetraazaannnulene pendants before the formation of the CS intermediate.

An effective identification and quantification method for Ginkgo biloba flavonol glycosides with targeted evaluation of adulterated products

BackgroundGinkgo biloba L. (Ginkgoaceae) leaf extract is one of the most popular herbal products on the market, as it contains flavone glycosides (≥ 24%) and terpene lactones (≥ 6%), which are proposed to have significant physiological effects. Unfortunately, the challenging financial climate has resulted in a natural health product market containing adulterated ginkgo products. Purpose42 ginkgo samples were analyzed to establish an HPLC profile for authentic ginkgo and common ginkgo adulterants, and to develop a method capable of easily detecting adulteration in ginkgo commercial products. MethodIn this study an efficient and targeted HPLC analysis method was established that is capable of distinguishing flavonol glycosides and aglycones simultaneously for the evaluation of ginkgo powdered extracts (PEs) and finished products in a single, 13min run. Thirteen ginkgo leaf samples, fifteen standardized powdered extracts, and fourteen commercially available ginkgo products have been analyzed using this new HPLC method. Chromatograms were compared to six standard reference materials: one flavonol glycoside (rutin), three aglycones (quercetin, kaempferol and isorhamnetin), and two isoflavones (genestin and genistein). The quantitative chromatographic data was interpreted by principal component analysis (PCA), which assisted in the detection of unexpected chromatographic features in various adulterated botanical products. ResultsOnly three of the commercially available ginkgo finished products tested in this study were determined to be authentic, with flavonol glycoside rutin, and aglycones quercetin, kaempferol, and isorhamnetin found to be common adulterants in the ginkgo powdered extract and finished product samples. ConclusionDespite evidence of adulteration in most of the samples, each of the samples discussed herein met most of the current pharmacopeial standards. It is therefore critical that a preliminary evaluation be utilized to detect adulteration in commercial ginkgo products, prior to the acid hydrolysis procedure utilized in the current testing methods.

Quercetin Stimulates Insulin Secretion and Reduces the Viability of Rat INS-1 Beta-Cells

Background/Aims: Previously we described insulinotropic effects of Leonurus sibiricus L. plant extracts used for diabetes mellitus treatment in Traditional Mongolian Medicine. The flavonoid quercetin and its glycoside rutin, which exert anti-diabetic properties in vivo by interfering with insulin signaling in peripheral target tissues, are constituents of these extracts. This study was performed to better understand short- and long-term effects of quercetin and rutin on beta-cells. Methods: Cell viability, apoptosis, phospho-protein abundance and insulin release were determined using resazurin, annexin-V binding assays, Western blot and ELISA, respectively. Membrane potentials (V_mem), whole-cell Ca²⁺ (ICa)- and ATP-sensitive K⁺ (IK_ATP) currents were measured by patch clamp. Intracellular Ca²⁺ (Ca_i) levels were measured by time-lapse imaging using the ratiometric Ca²⁺ indicator Fura-2. Results: Rutin, quercetin and the phosphoinositide-3-kinase (PI3K) inhibitor LY294002 caused a dose-dependent reduction in cell viability with IC₅₀ values of ∼75 µM, ∼25 µM and ∼3.5 µM, respectively. Quercetin (50 µM) significantly increased the percentage of Annexin-V+ cells within 48 hrs. The mean cell volume (MCV) of quercetin-treated cells was significantly lower. Within 2 hrs, quercetin significantly decreased basal- and insulin-stimulated Akt(T308) phosphorylation and increased Erk1/2 phosphorylation, without affecting P-Akt(S473) abundance. Basal- and glucose-stimulated insulin release were significantly stimulated by quercetin. Quercetin significantly depolarized V_mem by ∼25 mV which was prevented by the K_ATP-channel opener diazoxide, but not by the L-type ICa inhibitor nifedipine. Quercetin significantly stimulated ICa and caused a 50% inhibition of IK_ATP. The effects on V_mem, ICa and IK_ATP rapidly reached peak values and then gradually diminished to control values within ∼1 minute. With a similar time-response quercetin induced an elevation in Ca_i which was completely abolished in the absence of Ca²⁺ in the bath solution. Rutin (50 µM) did not significantly alter the percentage of Annexin-V+ cells, MCV, Akt or Erk1/2 phosphorylation, insulin secretion, or the electrophysiological behavior of INS-1 cells. Conclusion: We conclude that quercetin acutely stimulates insulin release, presumably by transient K_ATP channel inhibition and ICa stimulation. Long term application of quercetin inhibits cell proliferation and induces apoptosis, most likely by inhibition of PI3K/Akt signaling.

Polifenolen profil i antioksidantna aktivnost na ekstrakti ot maslinovi lista

The compounds, derivatives of olive leaves have a high antioxidant activity. The content of the total phenolic compounds (TPC), antioxidant activity (AOA) and HPLC polyphenol profile of methanol extracts from the leaves of the olive cultivars Chondrolia Halkidiki, Kalamon, Koroneiki grown in the nursery (in vivo) and in vitro plants of Chondrolia Halkidiki were compared. The results obtained for TPC varied between 9.2±0.5 mgGAE*gDW-1 and 16.4±0.5 mgGAE*gDW-1 . Antioxidant capacity was determined by four methods DPPH, ABTS, FRAP and CUPRAC. The highest results for TPC and AOA were achieved for the leaves of Chondrolia Halkidiki grown in vitro. A high correlation between the results gained from the TPC and AOA was established. Conducted HPLC analysis revealed the presence of 3,4- dihydroxybenzoic, caffeic, sinapic and ferulic acids and quercetin, hesperidin and luteolin and the quercetin glycosides rutin and hyperoside.

Protective effects of quercetin glycosides, rutin, and isoquercetrin against 6-hydroxydopamine (6-OHDA)-induced neurotoxicity in rat pheochromocytoma (PC-12) cells.

There is increasing evidence that free radicals induced oxidative stress is a major causative agent in the pathogenesis of neurodegenerative diseases, particularly Parkinson's disease. Quercetin glycosides, namely rutin and isoquercitrin, are flavonoid polyphenol compounds found ubiquitously in fruits and vegetables and have been known to possess antioxidant effects. This study was designed to compare the neuroprotective effects of quercetin glycosides rutin and isoquercitrin in 6-OHDA-induced rat pheochromocytoma (PC-12) cells. The results showed that both rutin and isoquercitrin significantly increased antioxidant enzymes, catalase, superoxide dismutase, glutathione peroxidase, and glutathione level that were attenuated by 6-OHDA in PC-12 cells. There was no significant difference in the activation of glutathione and glutathione peroxidase enzymes between rutin and isoquercitrin. These two glycosides were equally effective in suppressing lipid peroxidation in 6-OHDA-induced PC-12 cells as both compounds suppressed the malondialdehyde generation and prevented cell damage. In conclusion, quercetin glycosides rutin and isoquercetrin are having a significant neuroprotective effect against 6-OHDA toxicity in PC-12 cells.

Optimization of rutin isolation from Amaranthus paniculatus leaves by high pressure extraction and fractionation techniques

A procedure based on the application of pressurized liquid extraction (PLE) was employed to recover the major Amaranthus spp. flavonoid glycoside rutin from dried ground plant leaves. A central composite design (CCD) was used to optimize rutin extraction; three factors were considered: extraction solvent (water/ethanol ratio, from 0/100 to 100/0), extraction time (from 5 to 20min) and extraction temperature (from 50 to 200°C). Optimization of the three extraction parameters revealed that water/ethanol ratio in the solvent mixture was the most-significant factor influencing rutin yield, followed by extraction temperature. The total rutin recovery varied from 6.95 to 14.03g/kg dry matter (DM) depending on the experimental conditions tested. The optimum extraction conditions for the highest rutin content were 70/30 (v/v) water/ethanol ratio, at 188°C and 20min as static extraction time. In this case rutin recovery from the leaves (14.30g/kg DM) was 4-times higher compared to that attainable using conventional ultrasound assisted extraction (3.62g/kg DM) with methanol:ethanol mixture (90:10) as solvent. Rutin recovery from defatted and non-defatted seeds of amaranth under optimum PLE conditions was 35.3 and 41.1mg/kg DM, respectively, whereas using ultrasound assisted extraction 19.7 and 34.6mg/kg DM were obtained, respectively.Extracts obtained under optimum conditions were further fractionated by using supercritical antisolvent fractionation (SAF). Extracts of amaranth leaves containing 70% water were mixed with supercritical CO2 and separated into two fractions according to the molecular mass and solubility of the contained compounds. A central composite experimental design was applied for the determination of the best fractionation conditions to maximize rutin enrichment. Maximum rutin concentration in raffinate (22.57g/kg) was recovered at 15MPa using a feed mixture flow rate of 0.3mL/min.

A review on Ficus palmata (Wild Himalayan Fig)

Traditionally, various plants are used for treatment of many diseases. Ficus palmata is a herbaceous perennial plant belonging to the family Moraceae. It contains a very juicy fruit and is used for making various products such as squash, jam and jelly from this fruit. The fruits contain chiefly sugars and mucilage and are principally used as an item of diet in several cases of constipation and in the diseases of the lungs and the bladder. The ficus palmata plant is used in various diseases, e.g. gastrointestinal disorders, hypoglycemia, tumour, ulcer, diabetes, hyperlipidemia and fungal infections. Traditionally, stem latex is applied to extract spines deeply lodged in the flesh. The phytochemical screening of the Ficus palmata plant extracts showed the presence of alkaloids, tannins, flavonoids, terpenoids and cardiac glycosides and aerial parts of Ficus palmata utilizing liquid–liquid fractionation and different chromatographic techniques resulted in the isolation of a new isomer of psoralenoside namely, transpsoralenoside in addition to, one triterpene: germanicol acetate, two furanocoumarins: psoralene, bergapten, one aromatic acid vanillic acid and the flavone glycoside rutin. The ficus palmata fruit shows antioxidant activity using free radical scavenging and ferric reducing activities. The plant also shows in vitro antibacterial and antifungal activities of petroleum ether, chloroform, ethyl acetate, acetone, methanolic, ethanolic and water extracts. Fruit extract were analyzed against cervical cancer cell lines for antiproliferative activity while aqueous extract of Ficus palmata leaves showed dose dependant anticalcinogenic action. Ficus palmata total plant extract was found to show hepatoprotective, nephroprotective, antiulcer and anticoagulant activity.

Determination of Flavonoids in Stellaria by High-Performance Liquid Chromatography–Tandem Mass Spectrometry

The genus Stellaria (Caryophyllaceae) presents widely distributed plants often used in traditional medicine. Flavonoids are highly active plant secondary metabolites that may be involved in some of the effects of Stellaria plants. In this study, two new high-performance liquid chromatography–electrospray ionization tandem mass spectrometry (HPLC–ESI-MS-MS) methods were developed for the determination of flavonoids and isoflavonoids in plant material. The separations were performed on a reverse-phase C18 column with gradient elution using methanol and water with 0.5% acetic acid as the mobile phase. Multiple reaction monitoring was used for the tandem mass spectrometry detection and the two most intensive transitions were chosen for the identification of each analyte. The limits of detection and the limits of quantification were between 0.2 and 15.0 ng/mL and 0.6 and 50.0 ng/mL. The developed methods were successfully used for the analyses of four representatives of the genus Stellaria. All the studied herbs contained luteolin and its 7-O-glycosides, naringenin, kaempferol, quercetin, its glycoside rutin, apigenin, and its 7-O-glycoside. One coumarin, scopoletin, was also found. Isoflavones were primarily represented by genistein, genistin, and ononin. Some of the analytes were detected for the first time in Stellaria sp. The findings support that these methods are suitable for analyses of plant material.

Phytochemical and pharmacological study of Ficus palmata growing in Saudi Arabia

Phytochemical study of the aerial parts of Ficus palmata utilizing liquid–liquid fractionation and different chromatographic techniques resulted in the isolation of a new isomer of psoralenoside namely, trans-psoralenoside (5) in addition to, one triterpene: germanicol acetate (1), two furanocoumarins: psoralene (2), bergapten (3), one aromatic acid vanillic acid (4) and the flavone glycoside rutin (6). Structures of the isolated compounds were established through physical, 1D- and 2D-NMR and MS data. The total extract and fractions of the plant were examined in vivo for its possible effects as hepatoprotective, nephroprotective, antiulcer and anticoagulant activities in comparison with standard drugs. Hepatoprotective activity was assessed via serum biochemical parameters including aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma glutamyl transpeptidase (GGT), alkaline phosphatase (ALP) and total bilirubin. Tissue parameters such as non-protein sulfhydryl groups (NP-SH), malonaldehyde (MDA) and total protein (TP) were also measured. In addition to tissue parameters, nephroprotective effect was evaluated by measuring the serum levels of sodium, potassium, creatinine and urea. Histopathological study for both liver and kidney cells was also conducted. Antiulcer activity was explored by observing stomach lesions after treatment with ethanol. Whole blood clotting time (CT) was taken as a measure for the anticoagulant activity of the extract. Antioxidant activity of the total extract and fractions of the plant was measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH) method and ascorbic acid as standard.

Varietal Differences in the Flavonol Content of Mulberry (Morus spp.) Leaves and Genetic Analysis of Quercetin 3-(6-Malonylglucoside) for Component Breeding

The varietal differences in the flavonol glycosides rutin, isoquercitrin, kaempferol 3-(6-rhamnosylglucoside), quercetin 3-(6-malonylglucoside), astragalin, quercetin 3-(6-acetylglucoside), and kaempferol 3-(6-malonylglucoside) contained in mulberry leaves were elucidated. This information was used for breeding mulberry cultivars with a high concentration of functional components. The flavonol content, composition, and proportion in leaves varied widely. 'Kobuchizawa 1' had the highest level of total flavonols (1819 mg/100 g of dry weight), 5 times higher than that of 'Mikurasima 15' (393 mg/100 g of dry weight). Quercetin 3-(6-malonylglucoside) was the most abundant flavonol, although it was not found in all cultivars. Quercetin 3-(6-acetylglucoside) was only found in 'Keguwa'. From the quercetin 3-(6-malonylglucoside) content in crossbred offspring, malonyltransferase, an enzyme involved in quercetin 3-(6-malonylglucoside) synthesis, was acquired according to Mendelian inheritance. An offspring with a higher quercetin 3-(6-malonylglucoside) level than both parents was obtained from the crossing. This suggested that crossbreeding was effective for acquiring cultivars with a higher content of quercetin 3-(6-malonylglucoside).

Evaluation of reduction kinetics of 2,2-diphenyl-1-picrylhydrazyl radical by flavonoid glycoside Rutin in mixed solvent based micellar media

The work evaluates the reduction kinetics of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical by Rutin in different mixed solvent based micellar media comprising of various cationic, anionic, zwitterionic, nonionic and their binary mixtures using spectrophotometric, fluorimetric and tensiometric methods. The analysis of kinetic measurements in micellar systems was done using models proposed by Goupy et al. and Sendra et al. applicable to initial and overall DPPH reduction kinetics respectively. The rate constants observed in the methanol–water media containing micelles (except in cationic micellar medium) were significantly faster than those without micelles suggesting the use of such media as efficient systems for evaluation of DPPH activity in comparison with the pure organic solvents having environmental concern. Rate of DPPH reduction was dependent on nature as well as concentration of amphiphile used. The results were explained on the basis of compartmentalization and mode of interaction of Rutin with micellar nanostructures.

Preparation of quercetin and rutin-loaded ceramide liposomes and drug-releasing effect in liposome-in-hydrogel complex system

In this study, we developed a 2-step delivery system to enhance transdermal permeation of quercetin and its glycoside rutin, an antioxidant. Liposome-in-hydrogel complex systems were prepared by incorporating ceramide liposomes, which consist of biocompatible lipid membranes, into cellulose hydrogel. We evaluated the encapsulation efficiency, in vitro release behavior, and skin permeability of formulations that remained stable for over 3weeks. Rutin had greater encapsulation efficiency and better in vitro release properties than quercetin. However, quercetin demonstrated greater skin permeability than rutin. We also found that liposome-in-hydrogel complex systems (quercetin, 67.42%; rutin 59.82%) improved skin permeability of quercetin and rutin compared to control (phosphate buffer, pH 7.4) (quercetin, 2.48%; rutin, 1.89%) or single systems of hydrogel (quercetin, 31.77%; rutin, 26.35%) or liposome (quercetin, 48.35%; rutin, 37.41%). These results indicate that liposome-in-hydrogel systems can function as potential drug delivery systems to enhance transdermal permeation of the water-insoluble antioxidants quercetin and rutin.

An Open-Label Pilot Study of a Formulation Containing the Anti-Inflammatory Flavonoid Luteolin and Its Effects on Behavior in Children With Autism Spectrum Disorders

BackgroundAccumulating evidence suggests an association between autism spectrum disorders (ASD) and inflammation in brain regions related to cognitive function. The natural flavonoid luteolin has antioxidant, anti-inflammatory, mast cell–blocking, and neuroprotective effects. It was shown to improve cognitive performance in a mouse model of ASD, but its effect in humans has not been adequately studied. ObjectivesThe goal of this study was to assess the effectiveness and tolerability in white children with ASD of a dietary supplement containing 2 flavonoids (>95% pure), luteolin (100 mg/capsule, from chamomile) and quercetin (70 mg/capsule), and the quercetin glycoside rutin (30 mg/capsule) from the Sophora japonica leaf, formulated in olive kernel oil to increase oral absorption. MethodsFifty children (4–10 years old; 42 boys and 8 girls) with ASD were enrolled in a 26-week, prospective, open-label trial at the 2nd University Department of Psychiatry at “Attikon” General Hospital, Athens, Greece. Children were referred for the study by their respective physicians or came from the practice of the senior author. ASD diagnosis by clinical assessment was based on the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision, symptom list and corroborated by using the Autism Diagnostic Observation Schedule. The dose of the study formulation used was 1 capsule per 10 kg weight per day with food. The primary outcome measures were the age-equivalent scores in the Vineland Adaptive Behavior Scales domains. Secondary outcomes included the Aberrant Behavior Checklist, the Autism Treatment Evaluation Checklist, and the Clinical Global Impression–Improvement score. Data were measured at baseline, week 18, and week 26. Parents were interviewed for any possible improvements they noticed and instructed to report any unusual adverse events. ResultsA total of 40 children completed the protocol. There was a significant improvement in adaptive functioning as measured by using the VABS age-equivalent scores (8.43 months in the communication domain, 7.17 months in daily living skills, and 8 months in the social domain; P < 0.005), as well as in overall behavior as indicated by the reduction (26.6%–34.8%) in Aberrant Behavior Checklist subscale scores. Age, sex, and history of allergies had no effect on the results, whereas the initial level of functioning or difficulty did predict the final outcome in most of the measures used. There was a transient (1–8 weeks) increased irritability in 27 of the 50 participants. ConclusionsThese results are encouraging in that the combination of the flavonoids luteolin and quercetin seemed to be effective in reducing ASD symptoms, with no major adverse effects. ClinicalTrials.gov identifier: NCT01847521.

Antioxidants and Physical Integrity of Lipid Bilayers under Oxidative Stress

Giant unilamellar vesicles (GUVs of diameter 5-25 μm) of soy phosphatidylcholine (PC), resistant to intense light exposure (400-440 nm, ~15 mW·mm(-2)), underwent budding when containing chlorophyll a (Chla) in the lipid bilayer ([PC]:[Chla] = 1500:1). On the basis of image heterogeneity analysis using inverted microscopy, a dimensionless entropy parameter for the budding process was shown to increase linearly during an initial budding process. Lipophilic β-carotene (β-Car, [PC]:[β-Car] = 500:1) reduced the initial budding rate by a factor of 2.4, while the hydrophilic glycoside rutin ([PC]:[rutin] = 500:1) had no effect. Chla photosensitized oxidation of PC to form linoleoyl hydroperoxides, further leading to domains of higher polarity in the vesicles, is suggested to trigger budding. The average dipole moment (μ) of linoleic acid hydroperoxides was calculated using density functional theory (DFT) to have the value of 2.84 D, while unoxidized linoleic acid has μ = 1.86 D. β-Carotene as a lipophilic antioxidant and singlet-oxygen quencher seems to hamper oxidation in the lipid bilayers and delay budding in contrast to rutin located in the aqueous phase. The effect on budding of GUVs as a detrimental process for membranes is suggested for use in assays for evaluation of potential protectors of cellular integrity and functions under oxidative stress.

Hepatoprotective and Free Radical Scavenging Activities of Extracts and a Major Compound Isolated from the Leaves of Cineraria abyssinica Sch. Bip. exA. Rich.

In Ethiopian traditional medicine the aqueous decoction of the leaves of Cineraria abyssinica Sch. Bip. exA. Rich (Asteraceae) is used for the treatment of various ailments including liver diseases, however, to date, there appears to have been no scientific report on the phytochemistry and claimed hepatoprotective activity of the plant. The main purpose of this study was, therefore, to carry out hepatoprotective and antioxidant activities of the leaf extracts of C. abyssinica. Hepatoprotective activities of the aqueous and 80% methanolic extracts as well as the methanol fraction of the leaves of C. abyssinica were investigated against carbon tetrachloride-induced liver damage in rats. Intraperitoneal administration of 2 ml/kg of CCl4 (50% in liquid paraffin) significantly (p < 0.001) raised the plasma levels of alkaline phosphatase (ALP), alanine aminotransferase (ALT ) and aspartate aminotransferase (AST) in the toxin group compared with the values in the control group. Pretreatment of rats with 200 mg/kg of the aqueous, 80% methanol extracts and the methanol fraction reduced the toxin-induced rise in plasma ALP (65%, 75.4%, 85%), ALT (46.1%, 42.3%, 75%), and AST (58%, 98%, 79%), respectively. The standard drug, silymarin (100 mg/kg) reduced serum ALP (88%), ALT (92%), and AST (87.3%). Bioactivity-guided fractionation of the methanol fraction resulted in the isolation of the flavonol glycoside rutin, whose structure was assigned on the basis of spectroscopic methods. The results of biochemical analysis were further verified by histopatholgical examination of the liver, which showed improved architecture, absence of necrosis and a decrease in inflammation, compared with the findings in the toxin group of animals. Both the extracts and rutin showed potent 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activities. Acute toxicity studies showed that the total extracts of the plant are nontoxic up to a dose of 3 g/kg. The present study revealed for the first time the presence of a hepatoprotective and antioxidant phytochemical in the leaves of C. abyssinica that scientifically validates the traditional use of the plant and its potential for the treatment of liver disorders.

ChemInform Abstract: Regioselective Hydroxylation of Diverse Flavonoids by an Aromatic Peroxygenase.

Aromatic peroxygenases are extracellular fungal biocatalysts that selectively oxidize a variety of organic compounds. We found that the peroxygenase of the fungus Agrocybe aegerita (AaeAPO) catalyzes the H2O2-dependent hydroxylation of diverse flavonoids. The reactions proceeded rapidly and regioselectively yielding preferentially monohydroxylated products, e.g., from flavanone, apigenin, luteolin, flavone as well as daidzein, quercetin, kaempferol, and genistein. In addition to hydroxylation, O-demethylation of fully methoxylated tangeretin was catalyzed by AaeAPO. The enzyme was merely lacking activity on the quercetin glycoside rutin, maybe due to sterical hindrance by the bulky sugar substituents. Mechanistic studies indicated the presence of epoxide intermediates during hydroxylation and incorporation of H2O2-derived oxygen into the reaction products. Our results raise the possibility that fungal peroxygenases may be useful for versatile, cost-effective, and scalable syntheses of flavonoid metabolites.