Hydroxyl Radical Scavenging Effect Research Articles

Chemical Constituents, Antioxidative and Antimicrobial Activities of Essential Oil and Oleoresin of Tailed Pepper (Piper Cubeba L)

The antimicrobial and antioxidant potentials of hydrodistilled essential oil and oleoresin (obtained using acetone as a solvent) of tailed pepper were carried out by different techniques. The results obtained from antioxidant activity measurements of essential oil and oleoresin against mustard oil were measured for duration of 28 days in terms of peroxide, thiobarbituric acid, total carbonyl and p-anisidine values. The results obtained from butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) was similar to both the oleoresin and essential oil. In addition, the inhibitory action in linoleic acid system was studied by monitoring accumulation of peroxide concentration. The radical scavenging capacity of both essential oil and oleoresin on 2, 2'-diphenyl-1-picrylhydrazyl (DPPH) radical were (71.2%) and (69.77%) respectively at 25 µL/mL. It was relatively lower in comparison with synthetic antioxidants (BHA-96.41%; BHT- 95.91%). The results obtained from reducing power, chelating effect and hydroxyl radical scavenging effect was also supported the antioxidant of essential oil and oleoresin. The tailed pepper essential oil and oleoresin showed 100% mycelial zone inhibition against Penicillium viridicatum at 3000 and 2000 ppm respectively in the poison food method. It is interesting to note that the essential oil revealed 100% clear zone inhibition against Aspergillus flavus at all tested concentrations. The chemical characterization of tailed pepper essential oil by GC and GC-MS resulted in the identification of 44 components accounting for 97.8% of the oil.

Expression of the phycoerythrin gene of Gracilaria lemaneiformis (Rhodophyta) in E. coli and evaluation of the bioactivity of recombinant PE

Phycoerythrin (PE) is one of the most important proteins involved in light capturing during photosynthesis in red algae. Its potential biological activities had gained wide concerns. In the present study, tumor cytotoxic and hydroxyl radical assay were preformed to detect the bioactivity of recombinant PE. Recombinant plasmids pGEX-PE and pBGL were transformed into E. coli BL21 to make two recombinant strains BEX (pGEX-PE) and BGL (pBGL). PE expressing in BEX (pGEX-PE) was validated by SDS-PAGE and Western blotting analysis. SDS-PAGE analysis indicated that the PE-GST fusion protein was mostly inclusion bodies. Specific expression of PE was confirmed by Western blotting analysis. The recombinant E. coli BEX (pGEX-PE) cells were collected and sonicated. The supernatants were reserved for the tumor cytotoxic experiments. The result of tumor cytotoxic assay indicated that the supernatants containing PE had the activity of inhibiting the growth of Hela cells and with the increase of protein concentration, the inhibiting rate increased from 37.31% to 63.26%, which showed significant difference from the control. Hydroxyl radical scavenging effect was tested with supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates treated with sonication and heating. For the sonication samples, the scavenging rates of the supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates were significantly higher than the negative control BL21(pGEX-4T) (P<0.02), and the scavenging rates increased slowly following the increase of the protein content. For the heating samples, except for the 0.2 mg mL−1 BGL (pBGL) products, the scavenging effects of the supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates were stronger than that of negative control BL21(pGEX-4T). However, the effect intensity was not positively correlated with the increase of the protein concentration. Though a partially decreased hydroxyl radical scavenging activity was led by heating, the biological activity was still retained and conspicuous. This research showed that phycoerythrin protein expressing in E. coli has the potential medical and sanitarian value.

Sonophotocatalytic degradation of methyl orange by nano-sized Ag/TiO2 particles in aqueous solutions

Sonophotocatalytic behaviour of methyl orange (MeO) in aqueous solution illuminated by light generated by a xenon lamp was investigated. For all three kinds of photocatalysts: Degussa P25 (75% anatase, 25% rutile, with a surface area of 55.07m2/g), Yili TiO2 (mainly anatase, with a surface area of 10.45m2/g) and Ag/TiO2 (silver loaded on Yili TiO2), the degradation followed pseudo-first order kinetics. The results showed a synergistic effect between sonolysis and photocatalysis. Some parameters affecting the sonophotocatalytic degradation of MeO with nanoparticles Ag/TiO2 were determined. The results indicated that the degradation ratio of MeO increased with the increase of ultrasonic power. An optimum 60mg/L of Ag/TiO2 added to relatively low concentrations of MeO was proved to have the most effective degradation efficiency. The study on the effects of hydroxyl radical (OH) scavengers (i.e. mannitol and dimethyl sulfoxide) on the MeO degradation indicated that OH radicals played an important role during MeO degradation, which enhanced MeO to be completely decomposed.

Sclerotial metamorphosis in filamentous fungi is induced by oxidative stress

Sclerotium-forming filamentous fungi are of great agricultural and biological interest because they can be viewed as models of simple metamorphosis. They differentiate by asexually producing sclerotia but the processes involved in sclerotial metamorphosis were poorly understood. In 1997, it was shown for the first time that the sclerotial differentiation state in Sclerotium rolfsii concurred with increasing levels of lipid peroxides. This finding prompted the development of a theory supporting that sclerotial metamorphosis is induced by oxidative stress. Growth factors that reduce or increase oxidative stress are expected to inhibit or promote sclerotium metamorphosis, respectively. This theory has been verified by a series of published data on the effect of certain hydroxyl radical scavengers on sclerotial metamorphosis, on the identification and quantification of certain endogenous antioxidants (such as ascorbic acid, β-carotene) in relation to the fungal undifferentiated and differentiated states, and on their inhibiting effect on sclerotial metamorphosis as growth nutrients. In 2004-2005, we developed assays for the measurement of certain redox markers of oxidative stress, such as the thiol redox state, the small-sized fragmented DNA, and the superoxide radical. These new advances allowed us to initiate studies on the exact role of glutathione, hydrogen peroxide, and superoxide radical on sclerotial metamorphosis. The emerging data, combined with similar data from other better-studied fungi, allowed us to make some preliminary postulations on the ROS-dependent biochemical signal transduction pathways in sclerotiogenic filamentous fungi.

The ability of thiourea to scavenge hydrogen peroxide and hydroxyl radicals during the intra‐coronal bleaching of bloodstained root‐filled teeth

Hydrogen peroxide, an agent used in the intra-coronal bleaching of root-filled teeth for over a century, has been shown to diffuse from the pulp chamber to the outer root surface. Furthermore, it has been demonstrated that destructive hydroxyl radicals, the by-products of the bleaching process, have been detected on the external root surface. The control of such diffusion may be of importance in minimizing the risk of invasive cervical resorption (ICR) which has been linked to intra-coronal bleaching of discoloured root-filled teeth using hydrogen peroxide. The aims of the present in vitro study are to quantify the diffusion of hydrogen peroxide and hydroxyl radicals to the outer root surface following intra-coronal bleaching, and to evaluate the ability of thiourea incorporated into the bleaching protocol to scavenge residual hydrogen peroxide and hydroxyl radicals. Thirty-five single rooted premolar teeth with intact cementum at the cemento-enamel junction were used in this project. Thirty teeth were stained with red blood cells and root-filled with gutta-percha and AH26. The five unstained teeth were root-filled and constituted a negative control (Group 1). The stained teeth were divided equally into the following experimental groups and subjected to various intra-coronal bleaching regimes: Group 2--'walking bleach' with 20 microl 30 per cent w/w hydrogen peroxide; Group 3--20 microl 30 per cent w/w hydrogen peroxide and thermocatalytically activated; Group 4--20 microl acidified thiourea; Group 5--20 microl acidified thiourea and 20 microl 30 per cent w/w hydrogen peroxide; Group 6--20 microl acidified thiourea and 20 microl one per cent sodium hypochlorite; Group 7--20 microl acidified thiourea, 20 microl one per cent sodium hypochlorite and 20 microl 30 per cent w/w hydrogen peroxide. The reaction products of the bleaching process were quantified at the outer root surface using high performance liquid chromatography and electrochemical detection (HPLC-ECD). Results showed that hydrogen peroxide used alone in Groups 2 and 3 was able to be detected at the outer root surface in 100 per cent of the samples, and that the presence of the hydroxyl radical generated in both groups was detected in equal amounts (P < 0.05). When thiourea was incorporated into the bleaching protocols in Groups 5-7, it was shown to scavenge both hydrogen peroxide and hydroxyl radicals to a significant degree (P < 0.05). Acidulated thiourea is an effective scavenger of residual hydrogen peroxide and hydroxyl radicals generated during the intra-coronal bleaching of bloodstained root-filled teeth.

Extracts and Essential Oil of Ledum palustre L. Leaves and Their Antioxidant and Antimicrobial Activities

The in vitro antimicrobial and antioxidant activities of the essential oil and methanol extracts of Ledum palustre L. were investigated. Using GC-MS analysis, we identified 37 compounds in the essential oil, constituting 87.58% of the total oil. There are several monoterpenes, of which sabinene is the major compound (16～17%). There are several oxygenated monoterpenes of which terpinen-4-ol (7.6%) and myrtenal (3.5%) are the main constituents. β-Selinene, a-selinene, y-elemene, a-caryophyllene are the main sesquiterpenes (2～6% range). The oil strongly reduced the diphenylpicrylhydrazyl radical (IC??=1.56 ㎍/mL) formation and exhibited a hydroxyl radical scavenging effect in the Fe³?-EDTA-H₂O₂ deoxyribose system (IC??=2.7 ㎍/mL), and also inhibited the nonenzymatic lipid peroxidation of rat liver homogenate (IC??=13.5 ㎍/mL). The polar phase of the extract showed antioxidant activity. The oil showed antimicrobial activity against Streptococcus pneumoniae, Clostridium perfringens, Candida albicans, Mycobacterium smegmatis, Acinetobacter lwoffii and Candida krusei while the water-insoluble parts of the methanolic extracts exhibited slight or no activity. This study confirms that the essential oil of Ledum palustre L. possesses antioxidant and low antimicrobial properties in vitro.

Growth Characteristics, Bio-chemical Composition and Antioxidant Activities of Benthic Diatom Grammatophora marina from Jeju Coast, Korea

Benthic diatoms are known as a good food for shellfish in nature and in commercial hatchery of Jeju Island, Korea. Grammatophora marina is commonly found as dominant benthic micro-algae in Jeju coastal waters throughout the year. To know the best growth conditions of this species, culture was done in terms of three parameters; water temperature, salinity and nutrients. Each parameter was controlled by temperature of 15, 20 and 25°C; salinity of 25, 30 and 35 psu; and nutrient concentrations of 50, 100 and 200%. F/2 media was used with artificial seawater for the culture, which was continued for two weeks with L:D cycle 12:12 by using fluorescent light. Maximum specific growth rate was recorded 1.68 d ‐1 at temperature of 25°C with salinity of 35 psu and nutrient concentration of 200% on 6 th day during the culture period. Maximum biomass was also observed 4.9 × 10 5 cells mL ‐1 in the same condition. This species may belong to the euryhaline and eutrophic habitat with warm condition. For nutritional aspects of this species, protein, lipid and carbohydrate were measured. The value of protein, lipid and carbohydrate was 4.96%, 15.82% and 5.65%, respectively. The antioxidant activities of 80% methanolic extract were 46.7%, 23.7% and 23.8% on DPPH (1,1-Diphenyl-2-picrylydrazy) radical, superoxide anion radical and hydrogen peroxide scavenging, respectively. Percentage metal chelating activity was 81.2%. Enzymatic extracts of Alcalase and Ultraflow showed remarkable scavenging activities on DPPH radical (86.5% and 57.2%, respectively), and superoxide anion scavenging activities were 45.3% and 41.4% from Kojizyme and Viscozyme extracts, respectively. Extract of Protomex revealed 24.8% activity on hydrogen peroxide and Neutase showed 30.8% on hydroxyl radical scavenging effects. Celluclast and Viscozyme extracts showed 33.2% and 32.1% activities on nitric oxide scavenging, respectively, while Alcalase showed 61.5% on metal chelating. This species contains higher lipids among the biochemical compounds and higher metal chelating activities from both 80% methanolic and enzymatic extracts.

Effects of Nitrite and Nitrate on DNA Damage Induced by Ultraviolet Light

UV light is a major cause of human skin cancers. Nitrite and nitrate are well-known potential risk factors for gastric cancer. Little attention has been paid to the relationship between UV light and nitrite or nitrate on cancer. We examined the effects of nitrite and nitrate on the damage to nucleosides and DNA induced by UV light from a mercury lamp and by sunlight at neutral pH. A biologically relevant dose of nitrite and nitrate increased the generation of nucleobases and malondialdehyde on the reaction of nucleosides and DNA with UV light. The efficiency of nitrite enhancing the reaction was higher than that of nitrate at low doses. The contribution of the hydroxyl radical as the reactive species was suggested from the results of the inhibitory effects of hydroxyl radical scavengers. Nitrite and nitrate also enhanced the formation of nucleobases and malondialdehyde from DNA induced by sunlight. In the presence of 5 muM nitrite, the concentration in human skin cells, the product yields by sunlight were 5-10-fold greater than those in the absence of nitrite. The addition of 80 muM NO(3)(-), a concentration in human skin cells, also increased the yields significantly. Nitrite and nitrate may play a role in enhancing the genotoxic effects of UV light in humans.

Antioxidant Properties of Myrtilli Folium , Phaseoli Fructus Sine Seminibus and Drug Mixture Extracts

In vitro investigations for the antioxidant and free radical scavenging activity of Myrtilli folium-, Phaseoli fructus sine seminibus- and a drug mixture (Equiseti herba, Myrtilli folium, Phaseoli fructus sine seminibus, Urticae folium) extracts showed antioxidant (LPO inhibitory and chain-breaking antioxidant) activity and free radical (superoxide anion and hydroxyl radical) scavenging effect. The extracts inhibited lipid peroxidation induced enzymatically by adding NADPH and non-enzymatically by adding Fe2+ in brain microsomes and in brain homogenates, respectively. The extracts reduced the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH), which showed chain-breaking antioxidant activity. The extracts scavenged superoxide radicals (O2-·) by inhibiting the reduction of nitro blue tetrazolium evoked by phenazine methosulphate. In addition, the extracts inhibited Fenton-reagent (Fe2+ and H2O2) induced deoxyribose degradation, therefore, it was concluded that the extracts have hydroxyl radical (OH·) scavenging property.

Mechanism of site-specific DNA damage induced by ozone

Ozone has been shown to induce lung tumors in mice. The reactivity of ozone with DNA in an aqueous solution was investigated by a DNA sequencing technique using 32P-labeled DNA fragments. Ozone induced cleavages in the deoxyribose-phosphate backbone of double-stranded DNA, which were reduced by hydroxyl radical scavengers, suggesting the participation of hydroxyl radicals in the cleavages. The ozone-induced DNA cleavages were enhanced with piperidine treatment, which induces cleavages at sites of base modification, but the inhibitory effect of hydroxyl radical scavengers on the piperidine-induced cleavages was limited. Main piperidine-labile sites were guanine and thymine residues. Cleavages at some guanine and thymine residues after piperidine treatment became more predominant with denatured single-stranded DNA. Exposure of calf thymus DNA to ozone resulted in a dose-dependent increase of the 8-oxo-7,8-dihydro-2′-deoxyguanosine formation, which was partially inhibited by hydroxyl radical scavengers. ESR studies using 5,5-dimethylpyrroline- N-oxide (DMPO) showed that aqueous ozone produced the hydroxyl radical adduct of DMPO. In addition, the fluorescein-dependent chemiluminescence was detected during the decomposition of ozone in a buffer solution and the enhancing effect of D 2O was observed, suggesting the formation of singlet oxygen. However, no or little enhancing effect of D 2O on the ozone-induced DNA damage was observed. These results suggest that DNA backbone cleavages were caused by ozone via the production of hydroxyl radicals, while DNA base modifications were mainly caused by ozone itself and the participation of hydroxyl radicals and/or singlet oxygen in base modifications is small, if any. A possible link of ozone-induced DNA damage to inflammation-associated carcinogenesis as well as air pollution-related carcinogenesis is discussed.

Hydroxyl radical scavenging effects of guaiacol used in traditional dental pulp sedation: Reaction kinetic study

Guaiacol, which is a phenolic compound with a methoxy group and used in traditional dental pulp sedation, has the property of inducing cell proliferation. To clarify these mechanisms of guaiacol, this study examined the hydroxyl radical (*OH) scavenging effects of guaiacol in vitro. Generation of *OH was carried out by the Fenton reaction using mixture of ascorbic acid, H2O2, and Fe(III)-EDTA, and *OH was detected by measuring the *OH-mediated production of degradation products of deoxyribose, which reacts with 2-thiobarbituric acid (TBA) and is relatively stable for a long time. At concentrations of 10(-10) M to 10(-3) M, guaiacol inhibited the TBA reactive substance (TBA-RS) formation in a dose-dependent manner. Phenol and formaldehyde were also found to inhibit the TBA-RS formation, but their inhibitory activities were lower than that of guaiacol. The concentrations of guaiacol, phenol, and formaldehyde needed to cause 50% inhibition of TBA-RS formation were approximately 5 x 10(-6), 5 x 10(-5), and 2 x 10(-3) M, respectively. In this reaction system, guaiacol showed no chelating reaction with ferrous ion and did not directly react with H2O2. Guaiacol also exhibited radical scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) stable free radical, but its scavenging activity was lower than that toward *OH. These results suggest that guaiacol is a potent scavenger of reactive oxygen radicals and that its radical scavenging activity may be associated with its effect on cell proliferation.

Degradation of Reactive Azo Dyes by UV/H2O2: Impact of Radical Scavengers

Hydroxyl radical (•OH) scavenging effects of carbonate , bicarbonate and chloride (Cl−) ions on the performance of an ultraviolet light-hydrogen peroxide (UV/H2O2) advanced oxidation process were investigated using a reactive azo dye, C.I. Reactive Red 141 as the model compound. It was found that in the absence of scavengers, complete color removal was possible in 15 min under the experimental conditions employed. The rate of color decay was inhibited remarkably with increasing concentrations of bicarbonate and carbonate species, especially when the distribution of alkalinity favored the dominance of carbonate ions. It was further observed that chloride inhibited the rate only when it was between 100–1250 mM as Cl−, and was ineffective at higher concentrations.

5‐Hydroxytryptophan is a more potent in vitro hydroxyl radical scavenger than melatonin or vitamin C

Hydroxyl radicals are involved in direct damage of important biomolecules. Potent radical scavengers such as vitamin C and indoles of the tryptophan family can avert the potential damage. Melatonin and its precursor 5-hydroxytryptophan (5-HTP) were compared with water-soluble vitamin C. Different scavenger concentrations were measured in a steady-state luminol chemiluminescence system (SLCL-system) with combined Fe(II) chloride (0.1 mm) and hydrogen peroxide (1.0 mm) as hydroxyl radical generators. 5-HTP showed highest hydroxyl radical scavenging effects with a 50% inhibition concentration (IC50) of 1.8 microm. For vitamin C an IC50 of 12.7 microm was measured, whereas melatonin in pure demineralized water was much less efficient (IC50=724 microm). A comparison between melatonin in aqueous solution and melatonin in ethanol solution revealed that melatonin was significantly more effective in pure demineralized water.

Distinguishing the 5,5-dimethyl-1-pyrroline N-oxide (DMPO)-OH radical quenching effect from the hydroxyl radical scavenging effect in the ESR spin-trapping method

The spin-trapping method has been widely accepted to measure the hydroxyl radical scavenging activity in electron spin resonance (ESR) spectroscopy, however, the disappearance of the trapped signal of the 5,5-dimethyl-1-pyrroline N-oxide (DMPO)-OH radical may lead to an erroneous conclusion, as the decreased signal must result from the rapid quenching of the DMPO-OH formed. The DMPO-OH quenching activity of a compound can be measured by tracing the signal decay resulting from its addition after the cessation of DMPO-OH formation. It was found that Fe(II) ions could quench the DMPO-OH radical quite rapidly, and phosphate ions could enhance the DMPO-OH radical quenching activity of the Fe(II) ions.

Antioxidant Activities of the Chloroform Extract of Solanum trilobatum

A chloroform extract from the anticancer herb Solanum trilobatum (CST) (Nadkarni, 1979) was analyzed and compared with reference antioxidants for its in vitro antioxidative properties such as scavenging of α,α-diphenyl-β-picryl hydrazyl (DPPH) and superoxide radicals, protection to deoxyribose degradation, reducing power, as well as inhibition of lipid peroxidation. Preliminary phytochemical analysis of CST by silica gel thin-layer chromatography showed the presence of simple phenols, phenolic acids, isoflavones, xanthones, and lignans. The antioxidative effect of CST was found to be concentration dependent to a certain extent and then leveled off with further increase in concentration. The IC50 for each antioxidative reaction studied was calculated. When compared to the reference antioxidant butylated hydroxytoluene (BHT), CST exhibited less scavenging effect on DPPH radicals and reducing power but a better superoxide radical scavenging effect. From a comparison of the hydroxyl radical scavenging effect of CST with catechin, it seemed that CST was four-times more effective than catechin. CST was also able to prevent the formation of ·OH-induced malondialdehyde (MDA) in rat liver homogenate.

Resveratrol scavenges reactive oxygen species and effects radical-induced cellular responses

Scavenging or quenching of the reactive oxygen species (ROS) involved in oxidative stress has been the subject of many recent studies. Resveratrol, found in various natural food products, has been linked to decreased coronary artery disease and preventing cancer development. The present study measured the effect of resveratrol on several different systems involving the hydroxyl, superoxide, metal/enzymatic-induced, and cellular generated radicals. The rate constant for reaction of resveratrol with the hydroxyl radical was determined, and resveratrol was found to be an effective scavenger of hydroxyl, superoxide, and metal-induced radicals as well as showing antioxidant abilities in cells producing ROS. Resveratrol exhibits a protective effect against lipid peroxidation in cell membranes and DNA damage caused by ROS. Resveratrol was also found to have a significant inhibitory effect on the NF-κB signaling pathway after cellular exposure to metal-induced radicals. It was concluded that resveratrol in foods plays an important antioxidant role.

Differential roles of hydrogen peroxide and hydroxyl radical in cisplatin-induced cell death in renal proximal tubular epithelial cells

Reactive oxygen species (ROS) have been suggested as important mediators of cisplatin-induced acute renal failure in vivo. However, our previous studies have shown that cisplatin-induced cell death in vitro could not be prevented by scavengers of hydrogen peroxide and hydroxyl radical in rabbit renal cortical slices. This discrepancy may be attributed to differential roles of ROS in necrotic and apoptotic cell death. We therefore examined, in this study, the roles of ROS in necrosis and apoptosis induced by cisplatin in primary cultured rabbit proximal tubule. Cisplatin induced necrosis at high concentrations over a few hours and apoptosis at much lower concentrations over longer periods. Necrosis induced by high concentration of cisplatin was prevented by a cell-permeable superoxide scavenger (tiron), hydrogen peroxide scavengers (catalase and pyruvate), and antioxidants (Trolox and deferoxamine), whereas hydroxyl radical scavengers (dimethythiourea and thiourea) did not affect the cisplatin-induced necrosis. However, apoptosis induced by lower concentration of cisplatin was partially prevented by tiron and hydroxyl radical scavengers but not by hydrogen peroxide scavengers and antioxidants. Cisplatin-induced apoptosis was mediated by the signaling pathway that is associated with cytochrome c release from mitochondria and caspase-3 activation. These effects were prevented by tiron and dimethylthiourea but not by catalase. Dimethylthiourea produced a significant protection against cisplatin-induced acute renal failure, and the effect was associated with an inhibition of apoptosis. These results suggest that hydrogen peroxide is involved in the cisplatin-induced necrosis, whereas hydroxyl radical is responsible for the cisplatin-induced apoptosis. The protective effects of hydroxyl radical scavengers are associated with an inhibition of cytochrome c release and caspase activation.

Cytoprotective effects of rebamipide and carteolol hydrochloride against ultraviolet B-induced corneal damage in mice.

To analyze whether rebamipide (REB) and carteolol hydrochloride (CH) protect against UVB-induced corneal damage in mice. BALB/c mice topically pretreated with REB (1 and 10 mM) or CH (1, 10, and 100 mM) were exposed to ultraviolet (UV) B light at 416 micro W/cm(2). To evaluate corneal damage, mire irregularity was graded, and the haze index was estimated by using digitized corneal images. The formation of oxidized DNA in the corneal epithelium resulting from UVB exposure was estimated by using quantitative immunohistochemistry for 8-hydroxy-2-deoxyguanosine (8OHdG index). To analyze the mechanism of cytoprotection by REB and CH against UVB-induced cell damage, the UV absorption spectrum in these agents was evaluated by spectrophotometry, and their hydroxyl radical scavenging effect was evaluated by the electron spin resonance (ESR) spin trapping technique with Fenton system hydroxy radical generation. Seventy-two hours after UVB exposure, the severity of mire irregularity, haze index, and 8OHdG index were significantly lower in mice pretreated with 10 mM (P < 0.05, P < 0.05, and P < 0.01, respectively) of REB and in mice pretreated with 10 mM (P < 0.05, P < 0.01, and P < 0.01, respectively) and 100 mM (P < 0.01, P < 0.01, and P < 0.01, respectively) of CH compared with mice treated with vehicle. The absorption spectrum of REB overlapped with the UVB wavelength, and that of CH overlapped partially. The ESR spin signal corresponding to the hydroxyl radical was reduced by the addition of REB or CH. REB and CH attenuate UVB-induced corneal damage, which may be partly responsible for their sunscreening and hydroxyl radical scavenging effects.

Possible role of nicaraven in neuroprotective effect on hippocampal slice culture.

Nicaraven is an agent that is especially beneficial in vasospasm or brain damage caused by subarachnoid hemorrhage. It ameliorates neurological deficits of patients and protects the central nervous system from ischemia. We investigated the neuroprotective effect of nicaraven against oxygen-glucose deprivation (OGD) induced or N-methyl-D-aspartic acid (NMDA) induced hippocampal neuronal cell death in organotypic brain slice cultures. The effect of nicaraven on hippocampal neuronal injury was evaluated by inhibition of uptake of propidium iodide (PI) into dead cells. The results demonstrated that nicaraven protected neuronal cells from both OGD- and NMDA-induced cell death. While nicaraven has a strong hydroxyl radical scavenging effect, another radical scavenger, N-acetyl-L-cysteine (NAC), inhibited cell death only caused by OGD. In contrast, the poly(ADP-ribose) synthetase (PARS) inhibitors 3-aminobenzamide (3-AB) and theophylline protected cells from both OGD- and NMDA-induced cell death. Since nicaraven has an inhibitory effect in PARS, as well as a radical scavenging effect, these results suggest that inhibition of hippocampal cell death caused by NMDA may be attributable to PARS inhibition by nicaraven.

Antioxidant and antimicrobial activity of the essential oil and methanol extracts of Achillea millefolium subsp. millefolium Afan. (Asteraceae)

The in vitro antimicrobial and antioxidant activities of the essential oil and methanol extracts of Achillea millefolium subsp. millefolium Afan. (Asteraceae) were investigated. GC-MS analysis of the essential oil resulted in the identification of 36 compounds constituting 90.8% of the total oil. Eucalyptol, camphor, α-terpineol, β-pinene, and borneol were the principal components comprising 60.7% of the oil. The oil strongly reduced the diphenylpicrylhydrazyl radical (IC 50=1.56 μg/ml) and exhibited hydroxyl radical scavenging effect in the Fe 3+–EDTA–H 2O 2 deoxyribose system (IC 50=2.7 μg/ml). It also inhibited the nonenzymatic lipid peroxidation of rat liver homogenate (IC 50=13.5 μg/ml). The polar phase of the extract showed antioxidant activity. The oil showed antimicrobial activity against Streptococcus pneumoniae, Clostridium perfringens, Candida albicans, Mycobacterium smegmatis, Acinetobacter lwoffii and Candida krusei while water-insoluble parts of the methanolic extracts exhibited slight or no activity. This study confirms that the essential oil of Achillea millefolium possesses antioxidant and antimicrobial properties in vitro.