Antioxidant, free radical scavenging and anti-inflammatory effects of aloesin derivatives in Aloe vera.

Antioxidant components in Aloe vera were examined for lipid peroxidation using rat liver microsomal and mitochondrial enzymes. Among the aloesin derivatives examined, isorabaichromone showed a potent antioxidative activity. The DPPH radical and superoxide anion scavenging activities were determined. As one of the most potent components, isorabaichromone together with feruloylaloesin and p-coumaroylaloesin showed potent DPPH radical and superoxide anion scavenging activities. Electron spin resonance (ESR) using the spin trapping method suggested that the potent superoxide anion scavenging activity of isorabaichromone may have been due to its caffeoyl group. As A. vera has long been used to promote wound healing, the inhibitory effects of aloesin derivatives for cyclooxygenase (Cox)-2 and thromboxane (Tx) A 2 synthase were examined and the participation of p-coumaroyl and feruloyl ester groups in the aloesin skeleton was demonstrated. These findings may explain, at least in part, the wound healing effects of A.vera. Abbreviations. ADP:adenosine diphosphate ASA:ascorbic acid BHT:butylated hydroxytoluene BSA:bovine serum albumin DMPO:5,5-dimethyl-1-pyrroline N-oxide DPPH:1,1-diphenyl-2-picrylhydrazyl EDTA:edetic acid HEPES: N-(2-hydroxyethyl)-piperazine- N-2'-ethane-sulfonic acid NADH:reduced nicotinamide adenine dinucleotide NADPH:reduced nicotinamide adenine dinucleotide phosphate NBT:nitroblue tetrazolium Pg:prostaglandin SOD:superoxide dismutase TBA:thiobarbituric acid TCA:trichloroacetic acid XOD:xanthine oxidase

Multielement determination and speciation of major-to-trace elements in black tea leaves by ICP-AES and ICP-MS with the aid of size exclusion chromatography.

A multielement determination of major-to-trace elements in black tea leaves and their tea infusions was carried out by ICP-AES (inductively coupled plasma atomic emission spectrometry) and ICP-MS (inductively coupled plasma mass spectrometry). Tea infusions were prepared as usual tea beverage by brewing black tea leaves in boiling water for 5 min. About 40 elements in tea leaves and tea infusions could be determined over the wide concentration range in 8 orders of magnitude. The extraction efficiency of each element was estimated as the ratio of its concentration in tea infusions to that in tea leaves. From the experimental results for the extraction efficiencies, the elements in black tea leaves were classified into three characteristic groups: (i) highly-extractable elements (>55%): Na, K, Co, Ni, Rb, Cs and Tl, (ii) moderately-extractable elements (20-55%): Mg, Al, P, Mn and Zn, and (iii) poorly-extractable elements (<20%): Ca, Fe, Cu, Sr, Y, Zr, Mo, Sn, Ba and lanthanoid elements. Furthermore, speciation of major-to-trace elements in tea infusions was performed by using a combined system of size exclusion chromatography (SEC) and ICP-MS (or ICP-AES). As a result, many diverse elements were found to be present as complexes associated with large organic molecules in tea infusions.

Inhibition of mitochondrial lipid peroxidation by Bakuchiol, a meroterpene from Psoralea corylifolia.

Bakuchiol, a meroterpene isolated from Psoralea corylifolia, prevented mitochondrial lipid peroxidation. Inhibition of oxygen consumption originating in lipid peroxidation was time-dependent. Bakuchiol protected mitochondrial respiratory enzyme activities against both NADPH-dependent and dihydroxyfumarate-induced peroxidation injury. Bakuchiol was shown to be effective to protect mitochondrial functions against oxidative stress.

Protection of mitochondrial functions against oxidative stresses by isoflavans from Glycyrrhiza glabra.

Isoflavan derivatives, glabridin (1), hispaglabridin A (2), hispaglabridin B (3), 4'-Omethylglabridin (4) and 3'-hydroxy-4'-O-methylglabridin (5), isolated from Glycyrrhiza glabra, were investigated for their ability to protect liver mitochondria against oxidative stresses. Mitochondrial lipid peroxidation linked to respiratory electron transport and that induced non-enzymatically were inhibited by these isoflavans. Hispaglabridin A (2) strongly inhibited both peroxidations and 3'-hydroxy-4'-O-methylglabridin (5) was the most effective at preventing NADH-dependent peroxidation. 3'-Hydroxy-4'-O-methylglabridin (5) protected mitochondrial respiratory enzyme activities against NADPH-dependent peroxidation injury. Dihydroxyfumarate-induced mitochondrial peroxidation was also prevented by this isoflavan. Isoflavans from G. glabra were shown to be effective in protecting mitochondrial function against oxidative stresses.

Antimicrobial triterpenes from Ilex integra and the mechanism of antifungal action.

Antimicrobial triterpenes were isolated from the fruits of Ilex integra. Their structures were elucidated by spectral data and identified as rotundic acid (1), ulsolic acid (2) and peduncloside (3). Triterpene 1 showed significantly broad antimicrobial activity against bacteria, yeast and filamentous fungi. The antifungal activity of 1 was reversed by fatty acids. Cellular constituents leaked from Candida albicans cells incubated with triterpene 1. These results suggest that the antimicrobial activity of triterpenes in I. integra is due to a change of membrane permeability arising from membrane lipid alteration.

Mode of antibacterial action of retrochalcones from Glycyrrhiza inflata.

Licochalcone A-D and echinatin, retrochalcones isolated from the roots of Glycyrrhiza inflata, showed antimicrobial activity. Among them, licochalcone A and C had potent activity against some Gram-positive bacteria. These retrochalcones inhibited oxygen consumption in susceptible bacterial cells. The oxidation of NADH in bacterial membrane preparations was also inhibited by them. NADH-cytochrome c reductase was inhibited by licochalcones, while cytochrome c oxidase was not. NADH-CoQ reductase and NADH-FMN oxidoreductase were not inhibited. The site of respiratory inhibition of licochalcones was thought to be between CoQ and cytochrome c in the bacterial respiratory electron transport chain.

Antioxidative and superoxide scavenging activities of retrochalcones in Glycyrrhiza inflata.

Licochalcone A, B, C, D and echinatin, retrochalcones isolated from the roots of Glycyrrhiza inflata (licorice), along with an ordinary chalcone isoliquiritigenin, were assessed for their inhibitory activities on lipid peroxidation in various systems and radical scavenging activity. Among those tested, licochalcones B and D strongly inhibited superoxide anion production in the xanthine/xanthine oxidase system. These two compounds also showed potent scavenging activity on DPPH radical. Microsomal lipid peroxidation induced by Fe(III)-ADP/NAPDH was inhibited almost completely by 3 micrograms/ml of licochalcones B and D. Mitochondrial lipid peroxidation induced by Fe(III)-ADP/NADH was more sensitive to these retrochalcones; almost complete inhibition was observed at 10 micrograms/ml of all retrochalcones tested. Licochalcones B and D scavenged superoxide anion in microsome. Furthermore, these retrochalcones protected red cells against oxidative hemolysis. These phenolic compounds were shown to be effective to protect biological systems against various oxidative stresses.

An inhibitor of aldose reductase and sorbitol accumulation from Anthocepharus chinensis.

A flavonoid glycoside was isolated from Anthocepharus chinensis. Its structure was elucidated by spectral data and determined to be myricetin 3-O-(4"-acetyl)-alpha-fucoside. This flavonoid glycoside and its aglycone showed potent inhibition against rat and porcine lens aldose reductase. The flavonoid aglycone also inhibited sorbitol accumulation in human red blood cells.

Single-trial analysis of P3 in patients with generalized epilepsy.

The latencies and amplitudes of averaged P3, and the latencies, amplitudes and frequency components of single EEG responses to target tones were analyzed in 9 control subjects (CS group), 6 epileptics whose mean IQ was 100 (EP group) and 6 epileptics whose mean IQ was 52 (RE group), using an auditory oddball task. All of the subjects responded to the target tones correctly and there were no differences in the incidence of error in response to the target tones, or in the latencies and amplitudes of the averaged P3 among the three groups. However, the reaction times (RTs) in the RE group were significantly longer than those in the other groups (P < 0.05). Single EEG responses to target tone (single-trial ERPs) were classified into 2 types, those with and those without the P3 component. Type 1 had the P3 component and was observed in 42% of all of the responses in the RE group, significantly less than those in the CS (64%) and EP (61%) groups. The peak latencies of P3 in type 1 were similar among the three groups, but the amplitudes of P3 in type 1 in the RE group were significantly greater than those in the CS and EP groups. RTs in the RE group were significantly longer than those in the other groups, and had no correlation with the P3 latencies of type 1. There was little difference in the results of the frequency analysis among the three groups. These results suggest that all subjects in three groups recognized the target tones correctly, but they did not evaluate every target tone, since the incidence of P3 was almost 60% in the CS and EP groups, and 40% in the RE group. The characteristics of cognition and evaluation in three groups were the same, but the decrease in incidence of evaluation and the dissociation between the cognition and the response execution might be caused by impairment of the subject-environment contact mechanism, which resulted in the decrement of IQ in the RE group.

Antioxidative action of diterpenoids from Podocarpus nagi.

Diterpenoids, totarol (1), totaradiol (2), 19-hydroxytotarol (3), totaral (4), 4 beta-carboxy-19-nortotarol (5), sugiol (6), isolated from Podocarpus nagi, were evaluated as antioxidants. Microsomal lipid peroxidation induced by Fe(III)-ADP/NADPH and mitochondrial lipid peroxidation induced by Fe(III)-ADP/ NADH were inhibited by these terpenoids. They inhibited linoleic acid autoxidation but not generation of superoxide anion. Totarol (1) protected mitochondrial respiratory enzyme activities against NADPH induced oxidative injury. Totarane diterpenes from P. nagi were shown to be effective to protect biological systems and function against various oxidative stresses.

Antioxidative constituents in Heterotheca inuloides.

Sesquiterpenoids, 7-hydroxy-3,4-dihydrocadalin and 7-hydroxycadalin, and flavonoids, quercetin, kaempferol and their glycosides, isolated from Heterotheca inuloides (Asteraceae), a Mexican medicinal plant known as "arnica", were evaluated as antioxidants. These compounds showed potent scavenging activity on diphenyl-p-picrylhydrazyl (DPPH) radical. Microsomal lipid peroxidation induced by Fe(III)-ADP/NADPH was inhibited by both terpenoids and flavonoids, though only flavonoids possessed superoxide anion scavenging activity in microsome. Flavonoids also scavenged enzymatically and non-enzymatically generated superoxide anion. On the other hand, mitochondrial lipid peroxidation induced by Fe(III)-ADP/NADH was inhibited only by sesquiterpenoids. Furthermore, these terpenes protected mitochondrial enzyme activity against oxidative stress. These results showed that two types of antioxidants existed in the dried flower of H. inuloides and would contribute to protection of tissues against various oxidative stresses.

Antiperoxidative activity of neolignans from Magnolia obovata.

Honokiol and magnolol, neolignans in Magnolia obovata, have been evaluated as antioxidants. Microsomal lipid peroxidation induced by Fe(III)-ADP/NADPH and mitochondrial lipid peroxidation induced by Fe(III)-ADP/NADH were inhibited by these compounds. These neolignans protected mitochondrial respiratory chain enzyme activity against NADPH-induced peroxidative stress and protected red cells against oxidative haemolysis. The anti-oxidative activity of honokiol was more potent than that of magnolol. Neolignans in M. obovata were shown to be effective in protecting biological systems and functions against oxidative stress.

Cytotoxic and antioxidative sesquiterpenoids from Heterotheca inuloides.

Four sesquiterpenoids, beta-caryophyllene, beta-caryophyllene 4,5 alpha-oxide, 7-hydroxy-3,4-dihydrocadalin, and 7-hydroxycadalin, isolated from the dried flower of Heterotheca inuloides Cass. (Asteraceae), have been found to exhibit cytotoxic activity against several solid tumor cell lines. Among them, 7-hydroxy-3,4-dihydrocadalin and 7-hydroxycadalin have also been found to inhibit autoxidative and microsomal lipid peroxidation.

Antifungal activity from Alpinia galanga and the competition for incorporation of unsaturated fatty acids in cell growth.

An antimicrobial diterpene was isolated from Alpinia galanga in the screening for potentiators of phytochemical antibiotic action. The structure was elucidated by spectral data and identified as (E)-8 beta, 17-epoxylabd-12-ene-15, 16-dial (1). Diterpene 1 synergistically enhanced the antifungal activity of quercetin and chalcone against Candida albicans. Antifungal activity of 1 was reversed by unsaturated fatty acids. Protoplasts of C. albicans were lysed by 1. These results suggest that antifungal activity of 1 is due to a change of membrane permeability arising from membrane lipid alternation.

Antiperoxidative components in Thymus vulgaris.

A biphenyl compound, 3,4,3',4'-tetrahydroxy-5,5'-diisopropyl-2,2'-dimethylbiphenyl (1), and a flavonoid, eriodicytol (2), were isolated as antioxidative components from the leaves of Thymus vulgaris by bioassay-directed fractionation. These compounds inhibited superoxide anion production in the xanthine/xanthine oxidase system. Mitochondrial and microsomal lipid peroxidation induced by Fe(III)-ADP/NADH or Fe(III)-ADP/NADPH were also inhibited by these compounds. Compound 1 is an extremely potent antioxidant; complete inhibition was observed at 1 microM against both microsomal and mitochondrial peroxidation. Furthermore, compound 1 protected red cells against oxidative hemolysis. These phenolic compounds were shown to be effective to protect biological systems against various oxidative stresses.

Inhibition of aldose reductase by dihydroflavonols in Engelhardtia chrysolepis and effects on other enzymes.

Astilbin and neoastilbin, dihydroflavonol rhamnosides from Engelhardtia chrysolepis, showed potent inhibition of lens aldose reductase. Kinetic analysis showed astilbin exhibited uncompetitive inhibition against both dl-glyceraldehyde and NADPH. These taxifolin glycosides were selective inhibitors of aldose reductase with no inhibition of NADH oxidase.

Inhibition of lipid peroxidation by sesquiterpenoid in Heterotheca inuloides.

A sesquiterpenoid, 7-hydroxy-3,4-dihydrocadalin, isolated from a Mexican medicinal plant Heterotheca inuloides was evaluated as an antioxidant. This sesquiterpenoid inhibited mitochondrial and microsomal lipid peroxidation induced by Fe(III)-ADP/NADH or Fe(III)-ADP/NADPH. Furthermore, 7-hydroxy-3,4-dihydrocadalin protected red cells against oxidative haemolysis. This sesquiterpene was thus shown to be effective in protecting biological systems against oxidative stresses.

Mode of antibacterial action of totarol, a diterpene from Podocarpus nagi.

The antimicrobial mechanism of totarol was studied using Pseudomonas aeruginosa IFO 3080. This diterpene inhibited oxygen consumption and respiratory-driven proton translocation in whole cells, and oxidation of NADH in membrane preparation. NADH-cytochrome c reductase was inhibited by totarol while cytochrome c oxidase was not. NADH-DPIP reductase and NADH-CoQ reductase were also inhibited. The site of respiratory inhibition of totarol was thought to be near CoQ in the bacterial electron transport chain.

Effect of Polygonum hydropiper sulfated flavonoids on lens aldose reductase and related enzymes.

The sulfated flavonoids in Polygonum hydropiper showed potent inhibiton against lens aldose reductase. Among these flavonoids isorhamnetin 3,7-disulfate (5) was most potent. Kinetic analysis showed that 5 exhibited noncompetitive inhibition against both dl-glyceraldehyde and NADPH.

Inhibition of lipid peroxidation and superoxide generation by diterpenoids from Rosmarinus officinalis.

Four diterpenoids, carnosic acid (1), carnosol (2), rosmanol (3), and epirosmanol (4), were isolated as antioxidative agents from the leaves of Rosmarinus officinalis by bioassay-directed fractionation. These diterpenoids inhibited superoxide anion production in the xanthine/xanthine oxidase system. Mitochondrial and microsomal lipid peroxidation induced by NADH or NADPH oxidation were also completely inhibited by these diterpenes at the concentration of 3-30 microM. Furthermore, carnosic acid protected red cells against oxidative hemolysis. These phenolic diterpenes were shown to be effective to protect biological systems against oxidative stresses.