Model of singlet oxygen scavenging by α-tocopherol in biomembranes

Quenching of singlet molecular oxygen (${}^1\text{Δ}{}_{\mathrm{g}}\text{O}{}_2$) by α-tocopherol (I) involves the hydroxy function of the chromanol ring of I. In phosphatidylcholine (PC) uni- and multilamellar vesicles this structural element of I is localized at the interface polar headgroup/hydrophobic core. A dielectric constant of ? ～ 25 was determined for this special region of the PC bilayer. The ratio k_Q/k_R of rate constants of quenching processes (k_Q) and irreversible reactions (k_R) of I with ${}^1\text{Δ}{}_{\mathrm{g}}\text{O}{}_2$ increases with decreasing polarity of the solvent. In ethanolic solutions where ? = 25.5, k_Q/k_R is about 40. Extrapolation of these results to phospholipid bilayers suggests that at the nearness of the ester carbonyl oxygen of the PC fatty acid moieties, α-tocopherol can deactivate approximately 40 ${}^1\text{Δ}{}_{\mathrm{g}}\text{O}{}_2$ molecules before being destroyed. It is concluded that in vivo, one may expect to find a higher k_Q/k_R ratio if the chromanol ring of I hides within the more hydrophobic interiors of the membrane surface peptides.     

Synthesis and radical scavenging activity of phenol–imidazole conjugates

Novel hydroxylated benzylideneamino imidazole derivatives were synthesized and their radical scavenging activity was assessed against DPPH and hydroxyl radicals. In the DPPH assay, most of the synthesized compounds showed an IC₅₀ in the range 3.2 μM ⩽ IC₅₀ ⩽ 8.4 μM, lower than the reference compound trolox (IC₅₀ = 9.5 μM) or the parent aldehydes (5.4 μM ⩽ IC₅₀ ⩽ 11.6 μM). The activity depends mainly on the phenolic subunit (number and position of the hydroxyl groups) and the extent of conjugation with the imidazole ring. In the deoxyribose assay, all the compounds, including parent imidazoles and aldehydes, showed high activity against the hydroxyl radical and the ability to chelate iron ions. At 5 μM concentration, the compounds protected the deoxyribose from degradation by hydroxyl radical between 62% and 38%.     

How do roads affect the habitat use of an assemblage of scavenging raptors?

Scavengers may benefit from the availability of dead animals along roads that result from collisions with vehicles. However, roads are also considered risky places for many species. Animal habitat selection patterns usually balance energy intake with mortality risk. In this work we analyzed the foraging space use of an assemblage of diurnal scavenging raptors in relation to distance from roads in northwest Patagonia. We selected patches at different distances from roads, and placed a sheep carcass in each patch during the night (n = 18 carcasses in total). In general, carcasses near roads were detected by diurnal scavenging raptors much faster than those far from roads. Smaller raptors such as southern caracaras (Caracara plancus), chimango caracaras (Milvago chimango), and black vultures (Coragyps atratus), were commonly associated with roads both in terms of overall detections and scavenging activities. Southern and chimango caracaras proved to be very good at detecting carcasses, were faster to land in order to feed from them, and were found in greater numbers near roads than far from them. Even though Andean condors (Vultur gryphus) and black-chested buzzard-eagles (Geranoaetus melanoleucus) flew all over the area, they chose to feed far from roads. Our work emphasizes that some scavengers have taken advantage of the novel food resources provided by roads whereas others are reluctant to feed near them. Within a scenario of an increasing number of roads, some species can extend their distributions favoring competition and biotic homogenization processes within original communities. We highlight the importance of taking into account large flying scavengers in land-use planning. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Efficacy of HOCl scavenging by sulfur-containing compounds: antioxidant activity of glutathione disulfide?

Hypochlorous acid (HOCl) is a bactericidal compound formed by activated neutrophils during inflammation. Overproduction of HOCl causes damage to tissues at the site of neutrophil accumulation. The deleterious effects of excessive HOCl formation can be attenuated using antioxidants. Thiols and thioethers are known to be very effective HOCl scavengers. In the present study, the potency of several sulfur-containing compounds to protect acetylcholinesterase, glutathione S-transferase P1-1 (GST P1-1) and alpha1-antiproteinease against inactivation by HOCl was determined. Surprisingly, glutathione disulfide was an effective protector of acetylcholinesterase against hypochlorous acid. Glutathione disulfide did not provide protection for GST P1-1 and alpha1-antiproteinease against oxidative inactivation by HOCl. The implications of this finding are discussed.     

Mechanistic study of the structure–activity relationship for the free radical scavenging activity of baicalein

Density functional theory calculations were performed to evaluate the antioxidant activity of baicalein. The conformational behaviors of both the isolated and the aqueous-solvated species (simulated with the conductor-like polarizable continuum solvation model) were analyzed at the M052X/6-311 + G(d,p) level. The most stable tautomers of various forms of baicalein displayed three IHBs between O4 and OH5, O5 and OH6, and O6 and OH7. The most stable tautomer of the baicalein radical was obtained by dehydrogenating the hydroxyl at C6, while the most stable anion tautomer was obtained by deprotonating the C7 hydroxyl in gaseous and aqueous phases. The expected antioxidant activity of baicalein was explained by its ionization potentials (IPs) and homolytic O–H bond dissociation enthalpies (BDEs), which were obtained via the UM052X optimization level of the corresponding radical species. Heterolytic O–H bond cleavages (proton dissociation enthalpies, PDEs) were also computed. The calculated IP, BDE, and PDE values suggested that one-step H-atom transfer, rather than sequential proton loss–electron transfer or electron transfer–proton transfer, would be the most favorable mechanism for explaining the antioxidant activity of baicalein in the gas phase and in nonpolar solvents. In aqueous solution, the SPLET mechanism was more important.     

Reactivity of carbon nanotubes: Free radical generation or scavenging activity?

Carbon nanotubes (CNTs) currently attract intense research efforts because of their unique properties which make them suitable for many industrial applications. When inhaled, CNTs constitute a possible hazard to human health. Several studies have shown that when instilled in the lung of experimental animals, CNTs induced an inflammatory and fibrotic response similar to that caused by other toxic particles which might be the result of oxidative stress caused by particle- and/or cell-derived free radicals. There is, however, no direct experimental evidence of a capacity of carbon nanotubes to generate directly free radicals. Here we report that multiwall carbon nanotubes (MWCNT) in aqueous suspension do not generate oxygen or carbon-centered free radicals in the presence of H2O2 or formate, respectively, as detected with the spin-trapping technique. Conversely, we observed that, when in contact with an external source of hydroxyl or superoxide radicals, MWCNT exhibit a remarkable radical scavenging capacity. It is therefore possible that the inflammatory reaction reported in vivo must be ascribed to MWCNT features other than particle-derived free radical generation.     

Radical scavenging and anti-inflammatory properties of STW 5 (Iberogast®) and its components

A combination of ethanolic extracts from nine medicinal plants is successfully used in STW 5 (Iberogast^®) for treatment of gastrointestinal disorders. To elucidate possible modes of action, the focus of this study is on antioxidant properties of the phytomedicine STW 5. In fact, functional gastrointestinal diseases, such as non-ulcer dyspepsia (NUD) and irritable bowel syndrome, are often initiated by or correlated to inflammatory processes, where oxidants such as reactive oxygen species (ROS) play a crucial role. Prominent in vivo sources of ROS generation are represented by the enzymes xanthine oxidase (XOD) or myeloperoxidase (MPO). Applying these enzymes in models in vitro, we show that STW 5 and its components possess strong antioxidant activities. Depending on the model investigated, even pro-oxidant activites of single components of STW 5 could be observed. Interestingly, these effects were absent in STW 5, indicating cooperation between the components. Moreover, if one of the component extracts of STW 5 is omitted, the antioxidant activity is reduced. Thus we conclude that all the single extracts combined in STW 5 are of importance for the therapeutic effect, working in concert. The component of STW 5 performing best in vitro differed with the model investigated, respectively, with ROS and ROS generators. In the XOD system, the extracts of lemon balm leaf and peppermint leaf showed the best antioxidant result, whereas concerning MPO driven chlorination reactions, bitter candy tuft extract was the most efficient antioxidant. Best protection against peroxynitrite induced oxidation of methionine like sulfur-compounds exhibited the STW 5 components lemon balm leaf, Matricaria flower and peppermint leaf.     

Does scavenging extend the host range of entomopathogenic nematodes (Nematoda: Steinernematidae)?

Living and freeze-killed natural and laboratory hosts, with different susceptibility to entomopathogenic nematodes, were exposed to the larvae of Steinernema affine and Steinernema kraussei in two different experimental arenas (Eppendorf tubes, Petri dishes), and the success of the colonisation and eventual progeny production were observed. Both nematodes were able to colonise both living and dead larvae of Galleria mellonella (Lepidoptera) and adult Blatella germanica (Blattodea) even though the progeny production in dead hosts was lower on average. Living carabid beetles, Poecilus cupreus, and elaterid larvae (Coleoptera) were resistant to the infection, however, both nematodes were able to colonise and multiply in several dead P. cupreus and in a majority of dead elaterid larvae. By scavenging, EPNs can utilise cadavers of insects that are naturally resistant to EPN infection, and so broaden their host range.     

Effect of •OH scavenging action by non-SH-containing angiotensin converting enzyme inhibitor imidaprilat using microdialysis

We examined the effect of non-SH-containing angiotensin converting enzyme (ACE) inhibitor imidaprilat on hydroxyl radical (•OH) generation using microdialysis. Salicylic acid in Ringer's solution containing sodium salicylate (0.5 n mol μL⁻¹ min⁻¹) was infused directly through a microdialysis probe to detect the generation of •OH as reflected by the formation of 2,3-dihydroxybenzoic acid (DHBA) in the myocardium of anesthetized rats. We compared the ability of two non-SH-containing ACE inhibitors (imidaprilat and enalaprilat) with an -SH-containing ACE inhibitor (captopril) to scavenge the •OH. When iron (II) was administered to animals pretreated with these three ACE inhibitors, a decrease in 2,3-DHBA of all three compounds was observed, as compared with the iron (II) only-treated group. All three ACE inhibitors were able to scavenge •OH generated by the action of iron (II). However, imidaprilat is a free radical scavenger more potent than enalaprilat. These results suggested that ACE inhibitors are probably not only related to the presence of the SH radical.     

Is the endogenous peroxyl-radical scavenging capacity of plasma protective in systemic inflammatory disorders in humans?

Systemic inflammatory response syndrome (SIRS) in humans is associated with heightened intravascular oxidative stress. The clinical significance of plasma endogenous antioxidative capability in SIRS remains undetermined. Time-sequence changes of plasma total radical-trapping antioxidant parameter (TRAP) and its components were measured in 135 patients with various clinical conditions leading to SIRS. The results were correlated with clinical parameters. Plasma TRAP significantly depressed upon diagnosis of SIRS (SIRS vs. healthy subjects (n = 50), 605.7 +/- 20.4 vs. 803.4 +/- 30.8 microM Trolox equivalent, p <.001). In survivors (n = 86), TRAP declined further during the course of SIRS, followed by a mild recovery at the end of follow-up. General linear mixed model analysis revealed that uric acid, vitamin C, vitamin E and unidentified antioxidants contributed to most of the changes in TRAP (each factor p <.001). In nonsurvivors (n = 49), TRAP increased steadily until death, and the increase was predominantly the result of the increased contribution of bilirubin (p <.01). Higher TRAP levels were not correlated with diminished blood oxidants formation (r = -0.13, p.05), lower intensity of lipid peroxidation (r = 0.261, p <.05) or lesser disease severity of SIRS. The results do not support the hypothesis that the endogenous peroxyl radical scavenging ability of plasma plays a protective role in the course of SIRS.     

Antioxidant activity of β-blockers: An effect mediated by scavenging reactive oxygen and nitrogen species?

The therapeutic effects of beta-blockers are normally explained by their capacity to block the beta-adrenoceptors, however, some of the beneficial cardiovascular effects shown by this group of compounds have already been associated with the antioxidant properties that some of them seem to possess. The beta-blockers atenolol, labetalol, metoprolol, pindolol, propranolol, sotalol, timolol, and carvedilol were tested for their putative scavenging activity for ROS (O(2)(-), H(2)O(2), HO(.), HOCl, and ROO(.)) and RNS ((.)NO and ONOO(-)). Some of the studied compounds are effective ROS and/or RNS scavengers, these effects being possibly useful in preventing oxidative damage verified in hypertension as well as in other cardiovascular diseases that frequently emerge in association with oxidative stress.     

Vitamins, phytoplankton and bacteria: symbiosis or scavenging?

The conclusion that over 25% of global primary production dependson direct algal/bacterial symbiosis involving vitamin B₁₂ [Croftet al., (2005) Algae acquire vitamin B₁₂ through a symbioticrelationship with bacteria. Nature, 438, 90–93] is patentlyfalse, for it is based on a misconception of the probable levelof the vitamin B₁₂ requirement in marine pelagic algae. A reviewof the various published attempts at measuring this requirementsuggests that it is likely to be so low that oceanic and coastalconcentrations of the vitamin would usually be sufficient tosustain the populations that occur without the assistance ofdirect algal/bacterial symbiosis. The levels measured are discussedin relation to method (batch or continuous culture) and protocolsused. Requirement values considered by the author to be acceptablerange from 0.1 to 0.3 pM for the vitamin growth saturation constant(K_S) and from 30 to 100 µL algal biomass pmol^–1vitamin for the yield.     

Plastoquinol is more active than α-tocopherol in singlet oxygen scavenging during high light stress of Chlamydomonas reinhardtii

In the present study, we have performed comparative analysis of different prenyllipids in Chlamydomonas reinhardtii cultures during high light stress under variety of conditions (presence of inhibitors, an uncoupler, heavy water). The obtained results indicate that plastoquinol is more active than α-tocopherol in scavenging of singlet oxygen generated in photosystem II. Besides plastoquinol, also its oxidized form, plastoquinone shows antioxidant action during the stress conditions, resulting in formation of plastoquinone-C, whose level can be regarded as an indicator of singlet oxygen oxidative stress in vivo. The pronounced stimulation of α-tocopherol consumption and α-tocopherolquinone formation by an uncoupler, FCCP, together with the results of additional model system studies, led to the suggestion that α-tocopherol can be recycled in thylakoid membranes under high light conditions from 8a-hydroperoxy-α-tocopherone, the primary oxidation product of α-tocopherol by singlet oxygen.     

Hypertension: does impaired endothelium-dependent relaxation affect superoxide scavenging?

The purpose of this study was to investigate the effects of acute nitric oxide synthase inhibition on mean arterial blood pressure, oxidative stress markers such as plasma malondialdehyde (MDA) concentration, intracellular antioxidant enzyme activities such as copper-zinc superoxide dismutase (Cu/Zn SOD) and catalase and on trace elements important for activity and stability of Cu/Zn-SOD. Wistar-Kyoto rats (approx 150 g) (n=11) were treated with N ^ω-nitro-l-arginine methyl esther (l-NAME) (0.5 mg/mL) for 2 d. Age- and bodyweight-matched rats (n=10) were used for control group. Their systolic blood pressures and heart rates were recorded daily during the experimental period and also before their blood samples were drawn. Plasma MDA, plasma and red cell zinc and copper concentrations, and red cell Cu/Zn-SOD and catalase activities were determined. A progressive rise in systolic arterial blood pressure was observed compared to the control group (p<0.001). The heart rate of the experimental group was reduced on the third day (p<0.05). Plasma MDA concentration and red cell catalase activity were increased in the experimental group (p<0.001 and p<0.001, respectively). Plasma copper and red cell zinc concentrations were also increased significantly in the experimental group (p<0.001 and p<0.01, respectively). In conclusion, impairment in endothelium-derived relaxation altered mean arterial blood pressure, oxidant status, and trace element concentrations. Presented at the Advanced Course (sponsored by NATO-ASI, SFRR, FEBS, UNESCO-MCBN, IUBMB) “Free Radicals, Nitric Oxide, and Inflamation: Molecular, Biochemical, and Clinical Aspects,” Lara, Antalya, Turkey, September 23–October 3, 2001.     

Superoxide scavenging activity of plastoquinone derivative 10-(6′-plastoquinonyl)decyltriphenylphosphonium (SkQ1)

The plastoquinone derivative 10-(6′-plastoquinonyl)decyltriphenylphosphonium (SkQ1) has the ability to scavenge superoxide anion radical. This ability is manifested both in vitro and in vivo in experiments with the bacterium Escherichia coli. The protective effect of SkQ1 in vivo significantly exceeds that of ascorbic acid.     

Decreased plasma superoxide scavenging activity in immunological disorders — carboxyethylgermanium sesquioxide (Ge-132) as a promoter of prednisolone

Summary We investigated so-called superoxide scavenging activity (SSA) of plasma in patients with several immunological disorders, such as rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), polymyo-dermatomyositis (PM), progressive systemic sclerosis (PSS), myasthenia gravis (MG) and autoimmune thyroid disease (AT), using the electron paramagnetic resonance/spin trapping technique. Since carboxyethylgermanium sesquioxide, Ge-132, has been reported to modulate both the immune response and leukocyte functions, we have studiedin vivo effect of Ge-132 on plasma SSA and other laboratory parameters in these disorders. The plasma SSA was significantly lower in RA, SLE, PM and PSS, but not in MG and AT, as compared with that in healthy controls. An inverse correlation was observed between plasma SSA and parameters such as erythrocytes sedimentation rates, absolute number of leukocytes, C-reactive protein and serum globulin levels. Furthermore, plasma SSA was significantly decreased in rheumatoid factor-positive patients as compared to negative patients. No correlation was observed between plasma SSA and factors such as ages, sex of patients or the other laboratory parameters, such as serum albumin, triglyceride, cholesterol, hemoglobin and serum iron levels. Patients treated with prednisolone, especially ones with RA, showed an increase of plasma SSA. It appears that Ge-132 promotes prednisolone effects. Our results indicate that a decrease in plasma SSA is not disease specific, but inversely correlates with the severity and activity of inflammation. The methodology to measure plasma SSA presented in this work provides a helpful tool for determining the actual activity of the diseases as well asin vivo studies of antiinflammatory agents.     

3-methoxy aroylhydrazones – free radicals scavenging,anticancer and cytoprotective potency

Objective: This study aimed to determine the capability of newly designed 3-methoxy derivatives of salicylaldehyde benzoylhydrazone to influence the oxidative stress processes and to test their in vitro cytotoxicity.

Methods: We have used chemiluminescent and spectrophotometric model systems containing different types of reactive oxygen species (OH^●, OCl^─ and O₂^─●). The hydrazones effect on the viability of Hep-G2, HEK-293 and SH-SY5Y cell lines was determined via MTT assay.

Results: The comparative analysis of the C₅₀ values of the chemiluminescent investigation demonstrated moderate activity against the hydroxyl radicals (C₅₀?>?50?μmol/L) and remarkable reactivity in the systems containing a superoxide radical and a hypochlorous anion (C₅₀?● generation and consequent 2′-deoxyribose oxidative damage excluded the possibility of quenching effect and proved the direct interaction of the studied compounds with that generated in the system reactive oxygen species (ROS). The encapsulation of the studied derivatives into chitosan-alginate particles led to the protection and stabilization of their antioxidant activity as revealed by a one-month study using the ABTS ^●+ method. The cytotoxic study revealed less pronounced effects against the non-malignant cell line (HEK-293) compared to Hep-G2 and SH-SY-5Y cells.

Discussion: The incorporation of a hydroxyl group in the hydrazide part of a parent molecule which relates to better antioxidant effect in most of the studied systems is associated with higher IC₅₀ values in all cytotoxicity experiments and relates to the cytoprotective effect against N-methyl-D-aspartate-induced excitotoxicity in SH-SY5Y human neuronal cells.