Antioxidant activities of edible lichen Ramalina conduplicans and its free radical-scavenging constituents

The aim of this study was to evaluate the antioxidant properties of an edible lichen Ramalina conduplicans. The extract exhibited potent anti-linoleic acid peroxidation activity, free radical-scavenging activity, and reducing power. The total phenolic contents were found to be high in the extract. Activity-guided bioautographic thin layer chromatography (TLC) and HPLC identified sekikaic acid and homosekikaic acid as the main free radical-scavenging compounds in R. conduplicans extract (IC₅₀ [50% inhibition concentration] = 0.082 and 0.276 mg/ml, respectively). The results suggested that this edible lichen species have the potential to be utilized as food additives or as protective drugs.     

Antioxidant properties of minocycline: neuroprotection in an oxidative stress assay and direct radical-scavenging activity

Minocycline is neuroprotective in animal models of a number of acute CNS injuries and neurodegenerative diseases. While anti-inflammatory and anti-apoptotic effects of minocycline have been characterized, the molecular basis for the neuroprotective effects of minocycline remains unclear. We report here that minocycline and a number of antioxidant compounds protect mixed neuronal cultures in an oxidative stress assay. To evaluate the role of minocycline's direct antioxidant properties in neuroprotection, we determined potencies for minocycline, other tetracycline antibiotics, and reference antioxidant compounds using a panel of in vitro radical scavenging assays. Data from in vitro rat brain homogenate lipid peroxidation and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays show that minocycline, in contrast to tetracycline, is an effective antioxidant with radical scavenging potency similar to vitamin E. Our findings suggest that the direct antioxidant activity of minocycline may contribute to its neuroprotective effects in some cell-based assays and animal models of neuronal injury.     

Effects of copper ions on the free radical-scavenging properties of reduced gluthathione: implications of a complex formation

The interaction between glutathione (GSH) and copper ions was investigated in vitro to determine whether such interaction could affect the free-radical scavenging properties of the tripeptide. To this end, the bleaching (decrease in OD734 nm) of a coloured solution containing the stable free-radical cation ABTS+, (which results from the addition of thiols to such a solution) was employed as an in vitro indication of the ability of the tripeptide to scavenge free radicals. While GSH bleached concentration-dependently (1.0-7.5 gM) the ABTS+-containing solution, its prior incubation (5 microM) in the presence of Cu+1 or Cu+2 ions (1-7.5 M) led to a metal concentration-dependent decrease of the bleaching capacity. At a ratio equal to one (5 microM each), the bleaching capacity of the copper plus GSH mixture was 50% of that seen for GSH alone. Further additions of copper (reaching ratios up to 2) did not result in greater decreases in the GSH-bleaching capacity. Noteworthy at the ratio of onewas that the copper plus GSH solutions maintained their bleaching capacity despite the lack of any DTNB-reactivity, i. e., the complete absence of thiols in the mixture. Mixtures of increasing concentrations of a fixed ratio (equal to 2) of copper plus GSH, which were found not to exhibit any DTNB-reactivity, showed a linear and concentration-dependent increase in bleaching capacity. The bleaching capacity remained unaltered when TRIEN, EDTA or histidine were added to pre-incubated (1:1) mixtures of copper plus GSH. However, the incubation of copper with TRIEN or EDTA (but not histidine) prior to GSH addition, totally prevented the loss of the original GSH-bleaching capacity. The present data supports the formation of a copper-glutathione complex which is stable to the presence of some copper-chelators, lacks all thiol reactivity, but fully conserves the free-radical scavenging properties of GSH.     

Polyphenols from peanut skins and their free radical-scavenging effects

Separation of the water-soluble fraction of peanut skins led to the isolation of five proanthocyanidins. Based on the spectroscopic investigation and partial acid catalyzed degradation, their structures were determined to be epicatechin-(2beta-->O -->7, 4beta -->6)-[epicatechin-(4beta-->8)]-catechin (1), epicatechin-(2beta-->O -->7, 4beta-->8) epicatechin-(4beta-->8)-catechin-(4alpha-->8)-epicatechin (2), and procyanidins B2 (3), B3 (4) and B4 (5). The absolute configuration of the new compounds was determined from their circular dichroism curves and the (1)H NMR spectra of analysis of flavan-3-ols formed by thiolytic degradation of 1 and 2 in the presence of a chiral dirhodium complex (dirhodium tetra-(R)-(trifluoromethyl) phenyl acetate).     

Intracellular ROS protection efficiency and free radical-scavenging activity of curcumin

Curcumin has many pharmaceutical applications, many of which arise from its potent antioxidant properties. The present research examined the antioxidant activities of curcumin in polar solvents by a comparative study using ESR, reduction of ferric iron in aqueous medium and intracellular ROS/toxicity assays. ESR data indicated that the steric hindrance among adjacent big size groups within a galvinoxyl molecule limited the curcumin to scavenge galvinoxyl radicals effectively, while curcumin showed a powerful capacity for scavenging intracellular smaller oxidative molecules such as H₂O₂, HO^•, ROO^•. Cell viability and ROS assays demonstrated that curcumin was able to penetrate into the polar medium inside the cells and to protect them against the highly toxic and lethal effects of cumene hydroperoxide. Curcumin also showed good electron-transfer capability, with greater activity than trolox in aqueous solution. Curcumin can readily transfer electron or easily donate H-atom from two phenolic sites to scavenge free radicals. The excellent electron transfer capability of curcumin is because of its unique structure and different functional groups, including a β-diketone and several π electrons that have the capacity to conjugate between two phenyl rings. Therfore, since curcumin is inherently a lipophilic compound, because of its superb intracellular ROS scavenging activity, it can be used as an effective antioxidant for ROS protection within the polar cytoplasm.     

Spectroscopic properties and reactivity of free radical forms of A2E

A pyridinium bisretinoid (A2E) is the only identified blue-absorbing chromophore of retinal lipofuscin that has been linked to its aerobic photoreactivity and phototoxicity. Pulse radiolysis has been used to study both the one-electron oxidation and the one-electron reduction of A2E in aqueous micellar solutions. The reduction to the semireduced A2E (lambda(max) broad and between 500 and 540 nm) was achieved with formate radicals and the subsequent decay of A2E* was slow (over hundreds of milliseconds) via complex kinetics. The long lifetime of the A2E* should facilitate its reactions with other biomolecules. For example, with oxygen, the A2E* produced the superoxide radical anion with a rate constant of 3 x 10(8) M(-1) s(-1). The A2E was also reduced by the NAD radical, the corresponding rate constant being 2.3 x 10(8) M(-1) s(-1). Other experiments showed that the one-electron reduction potential of A2E lies in the range -640 to -940 mV. The semioxidized form of A2E (lambda(max) 590 nm) was formed via oxidation with the Br2*- radical and had a much shorter lifetime than the semireduced form. With strongly oxidizing peroxyl radicals (CCl3O2*) our kinetic data suggest the formation of a radical adduct followed by dissociation to the semioxidized A2E. With milder oxidizing peroxyl radicals such as that from methanol, our results were inconclusive. In benzene we observed an efficient oxidation of zeaxanthin to its radical cation by the A2E radical cation; this may be relevant to a detrimental effect of A2E in vision.     

Synthesis of new N-isobutyryl-L-cysteine/MEA conjugates: evaluation of their free radical-scavenging activities and anti-HIV properties in human macrophages

Four novel N-isobutyryl-l-cysteine/2-mercaptoethylamine (MEA, cysteamine) conjugates have been designed and synthesized. The antioxidant activities of these new series were evaluated by three different free radical scavenging methods (DPPH test, ABTS test, and deoxyribose assay) and their metal binding capacity was evaluated by the ethidium bromide fluorescence binding assay. These results were compared with those obtained with their pro-GSH acetyl analogues recently developed in our laboratory. We observed that most of these compounds exhibit free radical-scavenging activities similar to those of Trolox, but always superior than NAC. While none of these new derivatives had pro-GSH activities, they displayed anti-HIV properties in human monocyte-derived macrophages infected in vitro. The present study demonstrates that these new N-isobutyryl derivatives, which are expected to have a greater bioavailability than their acetyl analogues, may have useful applications in HIV infection in respect to their antioxidant and anti-HIV activities.     

Antioxidant properties of a human neuropeptide and its protective effect on free radical‐induced DNA damage

Human catestatin CgA_352–372 (SL21) is an endogenous neuropeptide with multiple biological functions. The present study aimed to evaluate the antioxidant, antibacterial, cytotoxic, and DNA damage protective effects of SL21 neuropeptide. SL21 neuropeptide generated from the C‐terminus of chromogranin A (CgA) was synthesized by solid‐phase method. Synthetic peptide was subjected to various in vitro antioxidant assays including the scavenging of 1,1‐diphenyl‐2‐pycryl‐hydrazyl (DPPH), 2,2‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS^·+), and hydroxyl free radicals, metal ion chelation, inhibition of lipid peroxidation, and reducing power. Moreover, protective effect of SL21 on H₂O₂‐induced DNA damage was analyzed using pTZ57/RT plasmid. Methylthiazoltetrazolium assay was also performed to study the cytotoxic effect of SL21 neuropeptide on human peripheral blood mononuclear cells. Furthermore, antibacterial and hemolysis assays were conducted. The results demonstrated high activities of SL21 in scavenging free radicals (DPPH, ABTS^·+, and hydroxyl), chelating of Cu²⁺/Fe²⁺ metal ions, reducing power, and inhibition of lipid peroxidation in a concentration‐dependent manner. SL21 neuropeptide revealed a protective effect on DNA damage caused by hydroxyl radicals. Interestingly, the peptide exhibited no significant cytotoxicity towards peripheral blood mononuclear cells. Furthermore, SL21 peptide displayed antimicrobial activity against Staphylococcus aureus and Pseudomonas aeruginosa without any hemolytic activity on human red blood cells. Conclusively, the present study established SL21 (catestatin) as a novel antioxidative peptide that could further be investigated for its potential use as a pharmaceutical agent. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd.     

Separation and free radical-scavenging activity of major curcuminoids of Curcuma longa using HPTLC-DPPH method

A direct HPTLC assay was developed for the determination of total curcuminoids and three individual curcuminoids, curcumin, demethoxycurcumin and bisdemethoxycurcumin. In addition, a new procedure was developed to separate and quantitative the free radical-scavenging activity of individual compounds from the rhizome of Curcuma longa L. (Zingiberaceae) based on the combination of HPTLC with a diode array detector (DAD) and post chromatographic DPPH(*) radical derivatisation. It was established that both individual curcuminoids and the extract of C. longa were capable of scavenging DPPH(*) radicals. From the estimated ID(50) values, it can be seen that the order of activity was curcumin > demethoxycurcumin > bisdemethoxycurcumin > ascorbic acid. However, the ID(50) values of curcuminoids were not significantly different. The data indicates the presence of a synergistic mechanism of antiradical activity of curcuminoids.     

Structure-activity relationship of C6-C3 phenylpropanoids on xanthine oxidase-inhibiting and free radical-scavenging activities

We employed the techniques of DNA relaxation, DPPH (1,1-diphenyl-2-picrylhydrazyl hydrate), and DMPO (5,5-dimethyl-1-pyrroline-N-oxide)-electron spin resonance (ESR), to study the effects of reactive oxygen species (ROS) suppression by 11 selected C6-C3 phenylpropanoid derivatives under oxidative conditions. We also investigated the effects of the derivatives on the inhibition of xanthine oxidase (XO) activity, and the structure-activity relationships (SARs) of these derivatives against XO activity were further examined using computer-aided molecular modeling. Caffeic acid was the most potent radical scavenger among the 11 test compounds. Our results suggest that the chemical structure and number of hydroxyl groups on the benzene ring of phenylpropanoids are correlated with the effects of ROS suppression. All test derivatives were competitive inhibitors of XO. The results of the structure-based molecular modeling exhibited interactions between phenylpropanoid derivatives and the molybdopterin region of XO. The para-hydroxyl of phenylpropanoid derivatives was pointed toward the guanidinium group of Arg 880. The phenylpropanoid derivatives containing the meta-or ortho-hydroxyl formed hydrogen bonds with Thr 1010. In addition, meta-hydroxyl formed hydrogen bonds with the peptide bond between the residues of Thr1010 and Phe1009. CAPE, the phenylenethyl ester of phenylpropanoids, had the highest affinity toward the binding site of XO, and we speculated that this was due to hydrophobic interactions of the phenylethyl ester with several hydrophobic residues surrounding the active site. The hypoxanthine/XO reaction in the DMPO-ESR technique was used to correlate the effects of these phenylpropanoid derivatives on enzyme inhibition and ROS suppression, and the results showed that caffeic acid and CAPE were the two most potent agents among the tested compounds. We further assessed the effects of the test compounds on living cells, and CAPE was the most potent agent for protecting cells against ROS-mediated damage among the tested phenylpropanoids.     

Antioxidant activity of flavonoids and reactivity with peroxy radical

The autoxidation of linoleic acid and methyl linolenate is inhibited by flavonoids. The antioxidant efficiency of these flavonoids increases with their concentration and in the order fustin < catechin < quercetin < rutin = luteolin < kaempferol < morin for linoleic acid and rutin < catechin < morin = kaempferol for methyl linolenate. Flavonoids are more effective on linoleic acid than on methyl linolenate. The antioxidant activity offlavonoids is related to an inhibition of the formation of trans,trans hydroperoxide isomers of linoleic acid. This inhibition exhibited the great H-atom donating ability of flavonoids to peroxy radical, thus terminating the chain radical reaction.     

Antioxidant, radical-scavenging, anti-inflammatory, cytotoxic and antibacterial activities of methanolic extracts of some Hedyotis species

The antioxidant, radical-scavenging, anti-inflammatory, cytotoxic and antibacterial activities of methanolic extracts of seven Hedyotisspecies were investigated. The antioxidant activity was evaluated by the ferric thiocyanate (FTC) and thiobarbituric acid (TBA) methods while the radical scavenging activity was measured by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. The anti-inflammatory activity related to NO inhibition of the plant extracts was measured by the Griess assay while cytotoxicity were measured by the MTT assay against CEM-SS cell line. The antibacterial bioassay (against 4 bacteria, i.e. Bacillus subtilis B28 (mutant), Bacillus subtilis B29 (wild-type), Pseudomonas aeruginosa UI 60690 and methicillin resistant Staphylococcus aureus, (MRSA) was also carried out using the disc-diffusion method. All tested extracts exhibited very strong antioxidant properties when compared to Vitamin E (alpha-tocopherol) with percent inhibition of 89-98% in the FTC and 60-95% in the TBA assays. In the DPPH method, H. herbacea exhibited the strongest radical scavenging activity with an IC50 value of 32 microg/ml. The results from the Griess assay showed that the tested extracts are weak inhibitors of NO synthase. However, all tested extracts exhibited moderate cytotoxic properties against CEM-SS cell line giving CD50 values in the range of 21-41 microg/ml. In the antibacterial bioassay, the stems and the roots of H. capitellata showed moderate activity against the 4 tested bacteria while the leaves showed moderate activity towards B. subtilis B28, MRSA and P. aeruginosa only. The roots of H. dichotoma showed strong antibacterial activity against all 4 bacteria. All other extracts did not exhibit any antibacterial activity.     

Pitfalls in limiting studies of reactivity to a family of compounds

Case studies of the oxidation potentials for MeCpMn(CO)₂B where B represents nitrogen and phosphorus donors, as well as the CO stretching frequencies of η-CpFe(CO)(COCH₃)PX₃ adducts are used to illustrate potential errors in data interpretation than can arise when studies are restricted to one family of compounds. These systems show that the uncertainty in the parameters from the data fit exceeds that indicated by R² or the standard deviations in the fit when the C_B/E_B ratios of the donors employed do not vary significantly. When an improper scale is used in data analyses, the R² for data fits is improved by limiting the correlation to a single family of compounds at the expense of meaningful interpretations. By definition, substituent constant correlations are limited to a single family, so care must be exercised in interpreting the parameters from these data fits.     

Antioxidant properties of a North American ginseng extract

A North American ginseng extract (NAGE) containing known principle ginsenosides for Panax quinquefolius was assayed for metal chelation, affinity to scavenge DPPH-stable free radical, and peroxyl (LOO·) and hydroxyl (·OH) free radicals for the purpose of characterizing mechanisms of antioxidant activity. Dissociation constants (Kd) for NAGE to bind transition metals were in the order of Fe²⁺ > Cu²⁺ > Fe³⁺ and corresponded to the affinity to inhibit metal induced lipid peroxidation. In a metal-free linoleic acid emulsion, NAGE exhibited a significant (p 0.05) concentration (0.01-10 mg/mL) dependent mitigation of lipid oxidation as assessed by the ammonium thiocyanate method. Similar results were obtained when NAGE was incubated in a methyl linoleate emulsion containing haemoglobin catalyst and assessed by an oxygen electrode. NAGE also showed strong DPPH radical scavenging activity up to a concentration of 1.6 mg/mL (r² = 0.996). Similar results were obtained for scavenging of both site-specific and non site-specific ·OH, using the deoxyribose assay method. Moreover, NAGE effectively inhibited the non site-specific DNA strand breakage caused by Fenton agents, and suppressed the Fenton induced oxidation of a 66 Kd soluble protein obtained from mouse brain over a concentration range of 2-40 mg/mL. These results indicate that NAGE exhibits effective antioxidant activity in both lipid and aqueous mediums by both chelation of metal ions and scavenging of free radicals.     

Synthesis and radical-scavenging activity of a dimethyl catechin analogue

Catechin analogue 1 with methyl substituents ortho to the catechol hydroxyl groups was synthesized to improve the antioxidant ability of (+)-catechin. The synthetic scheme involved a solid acid catalyzed Friedel–Crafts coupling of a cinnamyl alcohol derivative to 3,5-dibenzyloxyphenol followed by hydroxylation and then cyclization through an intermediate orthoester. The antioxidative radical scavenging activity of 1 against galvinoxyl radical, an oxyl radical, was found to be 28-fold more potent than (+)-catechin.     

Chemical composition and free radical-scavenging,anticancer and anti-inflammatory activities of the essential oil from Ocimum kilimandscharicum

ObjectiveThe essential oil from the leaves of Ocimum kilimandscharicum (EOOK), collected in Dourados-MS, was investigated for anticancer, anti-inflammatory and antioxidant activity and chemical composition.Materials and methodsThe essential oil was extracted by hydrodistillation, and the chemical composition was performed by gas chromatography–mass spectrometry. The essential oil was evaluated for free radical-scavenging activity using the DPPH assay and was tested in an anticancer assay against ten human cancer cell lines. The response parameter (GI₅₀) was calculated for the cell lines tested. The anti-inflammatory activity was evaluated using carrageenan-induced pleurisy in mice.ResultsThe chemical composition showed 45 components with a predominance of monoterpenes, such as camphor (51.81%), 1,8 cineole (20.13%) and limonene (11.23%). The EOOK exhibited potent free radical-scavenging activity by the DPPH assay with a GI₅₀ of 8.31 μg/ml. The major constituents, pure camphor (IC₅₀ = 12.56 μg/ml) and mixture of the limonene: 1, 8 cineole (IC₅₀ = 23.25 μg/ml) displayed a potent activity. The oral administration of EOOK (at 30 and 100 mg kg⁻¹), as well as the pure camphor or a mixture of 1,8 cineole with limonene, significantly inhibited the carrageenan (Cg) induced pleurisy, reducing the migration of total leukocytes in mice by 82 ± 4% (30 mg kg⁻¹ of EOOK), 95 ± 4% (100 mg kg⁻¹ of EOOK), 83 ± 9% (camphor) and 80 ± 5% (mixture of 1,8 cineole:limonene 1:1). In vitro cytotoxicity screening against a human ovarian cancer cell line displayed high selectivity and potent anticancer activity with GI₅₀ = 31.90 mg ml⁻¹. This work describes the anti-inflammatory, anticancer and antioxidant effects of EOOK for the first time.ConclusionsThe essential oil exhibited marked anti-inflammatory, antioxidant and anticancer effects, an effect that can be attributed the presence of majorital compounds, and the response profiles from chemical composition differed from other oils collected in different locales.     

Antioxidant properties of steroids

To determine the relative ranking of antioxidative potential of various steroids the effect of 14 steroid compounds on the fluorescence of phycoerythrin was monitored over time following the addition of a peroxy radical generator 2,2′-azobis (2-amidino-propane) dihydrochloride. The rate of decay of fluorescence in the presence of a 200 nM of 17β-estradiol, 17-estradiol and estriol expressed as percentages of the rate of decay in the absence of these compounds (control curve), were 74.1±6.3, 84.0±5.42 and 64.2±2.53%, respectively (P<0.005). Cortisone and corticosterone appeared to have very mild proxidant properties. Other steroids tested such as esterone, testosterone, progesterone, androstenedione, dehydroepiandrosterone, cortisol, tetrahydrocortisone, deoxycorticosterone and aldosterone had no significant antioxident properties. It is concluded that estrogens especially estriol and 17β-estradiol are naturally occurring antioxidants.     

Antioxidant properties of chromium and zinc

The effects of chromium (chromium picolinate, CrPic) and zinc (ZnSO₄H₂O) supplementation on serum concentrations of malondialdehyde (MDA) (an indicator of lipid peroxidation) and serum status of some antioxidant vitamins and minerals of laying hens (Hy-Line) reared at a low ambient temperature (6.8°C) were evaluated. One hundred twenty laying hens (Hy-Line; 32 wk old) were divided into 4 groups, 30 hens per group. The hens were fed either a basal diet or the basal diet supplemented with either 0.4 mg Cr/kg of diet, 30 mg Zn/kg of diet, or 0.4 mg Cr plus 30 mg Zn/kg of diet. Digestibility of nutrients (dry matter [DM], organic matter [OM], crude protein [CP], and ether extract [EE]) increased by supplementation of chromium and zinc (p<0.05). Supplemental chromium and zinc increased serum vitamins C and E but decreased MDA concentrations (p<0.05). Additionally, supplemental chromium and zinc caused an increase in the serum concentrations of Fe, Zn, Mn, and Cr (p < 0.05). The present study showed that low ambient temperature causes detrimental effects on the digestibility of nutrients and antioxidant status and that such detrimental effects caused by low ambient temperature can be alleviated by chromium and zinc supplementation, particularly when Cr and Zn were simultaneously included into the diet. Data obtained in the present study suggest that such supplementation can be considered as a protective management practice in a diet of laying hens for alleviating negative effects of cold stress.     

Antioxidant and free radical scavenging activities of an exopolysaccharide from a probiotic bacterium

Probiotic bacteria synthesize extracellular polysaccharides (EPSs) with commercially significant physiological and therapeutic activities. This important class of biomolecules is also characterized by their ability to remove reactive oxygen species (ROS) that are formed in the intestine by various metabolic reactions; hence, they exhibit antioxidant activities. Our probiotic bacterium, Bacillus coagulans RK-02, produces an EPS during the exponential and stationary growth phases when grown in a glucose mineral salts medium. The time course of EPS synthesis was studied with respect to biomass growth. The antioxidant and free radical scavenging potential of isolated EPS were studied by various methods, including the beta-carotene-linoleic acid model system, a superoxide radical scavenging assay using the PMS-NADH-nitroblue tetrazolium system, the 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, a hydroxyl radical scavenging assay using the ascorbic acid-Cu(2+)-cytochrome c system and an in vitro microsome peroxidation inhibition study using a thiobarbituric acid assay. The antioxidant activities were compared to known antioxidants vitamin C and E, which were used as reference standards. The results showed that the EPS, which is a heteropolymer composed of four monosaccharides, produced by B. coagulans RK-02 had significant antioxidant and free radical scavenging activities.     

Antioxidant properties of alpha-crystallin

-Crystallin is a major chaperone lens protein to which has been ascribed antioxidant functions. In the present work we have evaluated the antioxidant and free radical scavenging properties of bovine -crystallin in a series of in vitro models: zimosan-induced, luminol-enhanced chemiluminescence response of polymorphonuclear leukocytes, the autoxidation of brain homogenate, bleaching of 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)-derived radical cations, trapping of peroxyl radicals, and reactivity toward hypochloric acid. In all these systems, the reactivity of -crystallin is higher than or similar to that of bovine serum albumin. It is concluded that, given the high concentrations of -crystallin in the lenses, its capacity to interact with free radicals and to remove hypochlorous acid could contribute to the maintenance of the lens functionality.