Free radical-scavenging activity of sulfurous water investigated by electron paramagnetic resonance (EPR) spectroscopy.

The aim of the study was to explore the antiradical activity of sulfurous water, used for inhalatory therapy (characterized by the presence of sulfhydryl [HS]) by means of electron paramagnetic resonance (EPR) spectroscopy. The effects of sulfurous water corresponding to the concentrations from 16 down to 0.25 µg/mL of HS were tested by means of Fenton reaction (HO•), KO2-crown ether system (O2-•), and EPR of Tempol and of Fremy's salt radical. All of these assays were made using natural sulfurous water or degassed sulfurous water (no detectable HS) or reconstituted sulfurous water (degassed plus NaHS). The free radicals were significantly inhibited by natural water with HS concentrations ranging from 16 to 1 µg/mL for HO•, Tempol, and Fremy's salt, and O2-• was significantly inhibited from 16 and 2 µg/mL. The tests of degassed water did not reveal any significant differences from baseline values. The tests of reconstituted water led to significant results overlapping those obtained using natural water, thus confirming the importance of the presence of HS group (reductive activity). The positive effects of the activity of sulfurous thermal water is partially based on the patients' subjective sense of well-being and partially on symptomatic (or general) clinical improvements that are sometimes difficult to quantify. These findings indicate that, in addition to their known mucolytic activity and trophic effects on respiratory mucosa, the HS groups in sulfurous water also have antioxidant activity that contributes to the water's therapeutic effects on upper and lower airway inflammatory diseases.

How the atomic force microscope works?

This chapter aims at giving a quick but precise introduction of the atomic force microscope from the working principle point of view. It is intended to provide a useful starting point to those who first approach the instrument giving a general sketch of the working principles and technical implementations as well as last improvements. Subheading 1 is introductory: it gives an overview of what the instrument does and why it has been developed. Subheading 2 is focused on measurement ranges and on the comparison with scanning electron microscope (SEM) and transmission electron microscope (TEM) which have similar ranges and resolutions but different sample interactions and applications. Subheading 3 gives an overview of the working principles and the most diffused technical implementations on which most of the commercial microscopes rely, as we think it gives the useful base knowledge to understand possible applications, instrument capabilities, and results. In particular, technical improvements taking place over the past few years are highlighted. Despite of the simple and not very technical approach, it has a key importance in understanding concepts at the base of Chapter 3, which is, on the other side, useful for beginners and experienced users as well. Subheading 4 compares different instrument architectures and can, therefore, be useful for those who are going to choose an instrument having clear final applications. Latest solutions are once more highlighted. Subheading 5 gives an overview and some suggestions to start working, both in air and in liquid. Following the general philosophy of the book, it follows more an "how to do" concept than a general theoretical approach. Subheading 6 contains the future developments of the techniques.

Measurement methods in atomic force microscopy.

This chapter is introductory to the measurements: it explains different measurement techniques both for imaging and for force spectroscopy, on which most of the AFM experiments rely. It gives a general overview of the different techniques and of the output expected from the instrument; therefore it is, at a basic level, a good tool to properly start a new experiment. Concepts introduced in this chapter give the base for understanding the applications shown in the following chapters. Subheading 1 introduces the distinction between spectroscopy and imaging experiments and, within the last ones, between DC and AC mode. Subheading 2 is focused on DC mode (contact), explaining the topography and the lateral force channel. Subheading 3 introduces AC mode, both in noncontact and intermittent contact case. Phase imaging and force modulation are also discussed. Subheading 4 explains how the AFM can be used to measure local mechanical and adhesive properties of specimens by means of force spectroscopy technique. An overview on the state of the art and future trends in this field is also given.

Recognizing and avoiding artifacts in atomic force microscopy imaging.

Atomic force microscopy (AFM) measurements could be affected by different kinds of artifacts; some of them derive from the improper use of the instrument and can be avoided by setting the correct experimental parameters and conditions. In other cases, distortions of the images acquired by AFM are intrinsically related to the operating principle of the instrument itself and to the kind of interactions taken into account for the reconstruction of the sample topography. A perfect knowledge of all the artifacts that can perturb AFM measurements is fundamental to avoid misleading interpretations of the results. In this chapter, all the most common sources of artifact are presented, and strategies to avoid them are proposed.Subheading 1 is a brief introduction to the chapter. In Subheading 2, the artifacts due to the interactions between the sample and the AFM tip are presented. Subheading 3 is focused on the deformations due to the AFM scanner nonlinear movements. The interaction with the environment surrounding the instrument can affect the quality of the AFM results and the environmental instability are discussed in Subheading 4. Subheading 5 shows the effects of an incorrect setting of the feedback gains or other parameters. Subheading 6 aims on the artifacts that can be produced by the improper use of the image processing software. Subheading 7 is a short guide on the test that can be done to easily recognize some of the artifacts previously described.

Imaging bacterial shape, surface, and appendages before and after treatment with antibiotics.

Antibiotics are particular type of drugs that are able to interfere in different ways to the metabolic -pathways of bacteria. This causes also morphostructural alterations of cell wall and surface appendages (flagella, fimbriae or pili, and filaments).Atomic force microscopy (AFM) is extremely useful for analyzing the three-dimensional structure of the surface of biological specimens, particularly bacteria. A step-by-step AFM methodology to be applied to different type of bacteria is reported and visual examples of the action of antibiotics are shown. Although scanning electron microscopy is still frequently used, the introduction of the AFM technique offers substantial benefits in real quantitative data acquisition in three dimensions, minimal sample preparation times, flexibility in ambient operating conditions (i.e., no vacuum is necessary), and effective three-dimensional magnification at submicron level.

Thymol-induced alterations in Candida albicans imaged by atomic force microscopy.

Thymol, a constituent of thyme essential oil that has been credited with interesting antimicrobial and antifungal effects, acts by interfering with the envelope of Candida albicans and this activity has been investigated by means of atomic force microscopy (AFM). Candida culture samples incubated with 1, 1/2, and 1/4 MIC of thymol or vehicle were taken at time 0 and after 1, 2, and 4 h, the envelopes of 100 cells in each of five randomly chosen fields were analysed by means of AFM. Our AFM findings show that thymol affects the envelope of C. albicans cells. The cells showed major morphostructural deformities with envelope damage becoming greater at increasing thymol concentrations and longer times of incubation, including the number of flattened cells with surface folds, cells with holes, and collapsed cells and ghosts. Thymol is an amphipathic monoterpene, which suggests that it affects cell membrane structure by generating asymmetries and membrane tensions. This is confirmed by the fact that terpenes alter cell permeability by entering between the fatty acyl chains making up the membrane lipid bilayers, disrupting lipid packing, and changing membrane fluidity. All of these phenomena lead to major surface alterations and deformities that also reduce the ability of fungi to adhere to mucosal cells, and decrease their virulence and infectiousness.

Protective effect of erdosteine metabolite I against hydrogen peroxide-induced oxidative DNA-damage in lung epithelial cells.

It has been shown that the mucolytic agent erdosteine (N-carboxymethylthio-acetyl-homocysteine thiolactone, CAS 84611-23-4) has anti-inflammatory and anti-oxidant properties, and an active metabolite I (MET I) containing pharmacologically active sulphydryl group has been found to have a free radical scavenging activity. The aim of this study was to assess the ability of erdosteine metabolite I to protect A549 human lung adenocarcinoma cell against hydrogen peroxide (H2O2)-mediated oxidative stress and oxidative DNA damage. When A549 cells were pre-treated with the active metabolite I (2.5-5-10 microg/ml) for 10-30 min and then exposed to H2O2 (1-4 mM) for two additional hours at 37 degrees C, 5% at CO2, the intracellular peroxide production, reflected by dichlorofluorescein (DCF) fluorescence, decreased in a concentration-dependent manner. Furthermore, using a comet assay as an indicator for oxidative DNA damage, it was found that the metabolite I prevented damage to cells exposed to shortterm H2O2 treatment. The data suggest that this compound is effective in preventing H2O2-induced oxidative stress and DNA damage in A549 cells. The underlying mechanisms involve the scavenging of intracellular reactive oxygen species (ROS).

Inhibitory activity of thymol on native and mature Gardnerella vaginalis biofilms: in vitro study.

Bacterial vaginosis (BV) is the most frequent diagnosis made in women with lower genital tract symptoms. It has recently been observed that 90 % of subjects with BV show the growth of bacteria in the form of biofilms as against only 10% without BV, and that Gardnerella vaginalis was the predominant species. The propensity of G. vaginalis to form biofilm is clinically relevant because this form of growth allows it to tolerate higher concentrations of certain antibiotics, thus increasing the possibilty of recurrent BV even after apparently curative therapy. The aim of this study was to investigate whether thymol (CAS 89-83-8), a molecule present in thyme essential oil, that is credited with having a series of pharmacological properties including antimicrobial and antifungal effects, can interfere with newly formed and mature G. vaginalis biofilms. The ability of G. vaginalis ATCC 49145 and two G. vaginalis strains isolated from human BV to form biofilm in flat-bottomed 96-well microtitre plates was verified, and the effects of thymol concentrations ranging from 1 to 1/16 MIC (minimum inhibitory concentration) on preformed and mature biofilms was investigated by means of spectrophotometric analysis, Nomarski interference contrast microscopy, and fluorescence microscopy with live-dead cell visualisation (SYTO 9 and propidium iodide). Native biofilm was inhibited by concentrations ranging from 1 MIC to 1/8 MIC (32.77% +/- 2.37 to 11.39% +/- 1.46), and mature biofilm was inhibited by concentrations ranging from 1 MIC to 1/4 MIC (26.18% +/- 1.36 to 13.20% +/- 1.44). Nomarski interference contrast and fluorescence microscopy visually confirmed these findings. As biofilm is a multi-factorial phenomenon, the multiple mechanisms of thymol may act on different steps in the evolution of mature biofilm.

Effects of sulphurous water on human neutrophil elastase release.

BACKGROUND: Molecules bearing a sulphide (HS) group, such as glutathione, play a fundamental role in the defensive system of human airways, as shown by the fact that the lining fluid covering the epithelia of the respiratory tract contains very high concentrations of glutathione: the lungs and nose, respectively, contain about 140 and 40 times the concentrations found in plasma. Consequently, various low-weight soluble molecules bearing an HS group (including N-acetylcysteine, mesna and thiopronine, and prodrugs such as stepronine and erdosteine) have been used for therapeutic purposes. HS groups can also be therapeutically administered by means of sulphurous thermal water containing HS groups. The aim of this study was to investigate the direct activity of such water on the release of elastase by activated human neutrophils. METHOD: After the neutrophils were incubated with increasing amounts of sulphurous water or the HS/hydrogen sulphide donor sodium hydrosulphide (NaHS), elastase release was initiated by N-formyl-methionyl-leucyl-phenylalanine and measured by means of spectrofluorimetry using methylsuccinylalanylprolylvalyl-methylcoumarin amide as the fluorogenic substrate. To verify the presence of direct action on elastase we determined the diameter of the area of elastinolysis on elastine-agarose gel plates. RESULTS: The sulphurous water significantly inhibited elastase release at HS concentrations ranging from 4.5 to 18 µg/ml, as assayed using the iodometric method; in the case of NaHS, the inhibition was significant at HS concentrations ranging from 2.2 to 18 µg/ml. The concentration-effect regression lines of both were parallel and neither showed any direct elastolytic activity. CONCLUSIONS: Previous claims concerning the activity of sulphurous water have been based on the patients' subjective sense of wellbeing and on symptomatic (or general) clinical improvements that are not easy to define or quantify exactly. Our findings indicate that, in addition to its known mucolytic and antioxidant activity, sulphurous water also has an anti-elastase activity that may help to control the inflammatory processes of upper and lower airway diseases.

Free radical scavenging activity of erdosteine metabolite I investigated by electron paramagnetic resonance spectroscopy.

The aim of this study was to explore the antiradical activity of Met I (an active metabolite of erdosteine) containing a pharmacologically active sulphydryl group, by means of electron paramagnetic resonance (EPR) spectroscopy which has not previously been used to characterize the antiradical activity of Met I. The effects of concentrations of 20, 10, 5, 2.5, 1.25 and 0.625 microg/ml of Met I were tested against: (a) the Fenton reaction model system with EPR detection of HO.; (b) the KO2-crown ether system with EPR detection of O2-.; (c) the EPR assay based on the reduction of the Tempol radical, and (d) the EPR assay based on the reduction of Fremy's salt radical. Our findings show that the intensity of 4 different free radicals was significantly reduced in the presence of Met I, thus indicating the presence of a termination reaction between the free radicals and Met I.

Thermal treatment of eggplant (Solanum melongena L.) increases the antioxidant content and the inhibitory effect on human neutrophil burst.

The aim of this study was to compare the amount and activity of phytonutrients in raw, grilled, and boiled eggplant fruit using chemical measures and a biological assay of oxidative bursts in human neutrophils. The thermally treated samples showed various changes in their chemical composition (dry matter, soluble solids, acidity, and the amount of alcohol insoluble substances) due to the cooking processes and were much richer in the main phenolic compounds such as chlorogenic and caffeic acids, which are known to be antioxidants. Consequently, their free radical scavenging activity was significantly higher, especially that of superoxide anion. The biological assay of oxidative bursts from human neutrophils in the presence of N-formyl-methionyl-leucyl-phenylalanine confirmed the greater activity of extracts of the cooked eggplants with respect to raw eggplants. Successive extract dilutions showed a significant activity up to 1.25 microg/mL after cooking, while raw fruits resulted in an activity up to 10.00 microg/mL. These results showed that the thermal treatment commonly used before consumption can increase the content and biological activity of antioxidant compounds of eggplants.

Vaginal gel adsorption and retention by human vaginal cells: visual analysis by means of inorganic and organic markers.

To improve efficiency and prolong protection, modern gynecological preparations frequently incorporate polymeric molecules that add a certain degree of viscosity in order to increase adhesion with vaginal cells and prolong local delivery of active molecules. The aim of this study was to investigate the possibility of visualising the ability of a commercial medicated gynecological gel to bind to and be retained by human vaginal cells. The gel formulation included the essential oils of Thymus vulgaris and Eugenia cariophylla, which contain active molecules such as thymol and eugenol that are known to have useful antibacterial and antimycotic activities. The adherence of different dilutions of the gel to human vaginal cells was visualised by means of Nomarski interference contrast microscopy and scanning electron microscopy using ferric oxide particles and Escherichia coli as inorganic and organic markers, both of which made it possible to visualise the binding of the thin transparent layer of gel and the retaining effect, which was proportional to the degree of dilution.

Antioxidant activity of Calendula officinalis extract: inhibitory effects on chemiluminescence of human neutrophil bursts and electron paramagnetic resonance spectroscopy.

There is growing interest in natural chemical compounds from aromatic, spicy, medicinal and other plants with antioxidant properties in order to find new sources of compounds inactivating free radicals generated by metabolic pathways within body tissue and cells, mainly polymorphonuclear leukocytes (PMNs) whose overregulated recruitment and activation generate a large amount of reactive oxygen species (ROS) and reactive nitrogen species (RNS), leading to an imbalance of redox homeostasis and oxidative stress. The aim of this study was to examine whether a propylene glycol extract of Calendula officinalis interferes with ROS and RNS during the PMN respiratory bursts, and to establish the lowest concentration at which it still exerts antioxidant activity by means of luminol-amplified chemiluminescence. Electron paramagnetic resonance (EPR) spectroscopy was also used in order to confirm the activity of the C. officinalis extract. The C. officinalis extract exerted its anti-ROS and anti-RNS activity in a concentration-dependent manner, with significant effects being observed at even very low concentrations: 0.20 microg/ml without L-arginine, 0.10 microg/ml when L-arginine was added to the test with phorbol 12-myristate 13-acetate and 0.05 microg/ml when it was added to the test with N-formyl-methionyl-leucyl-phenylalanine. The EPR study confirmed these findings, 0.20 microg/ml being the lowest concentration of C. officinalis extract that significantly reduced 2,2-diphenyl-1-picrylhydrazyl. These findings are interesting for improving the antioxidant network and restoring the redox balance in human cells with plant-derived molecules as well as extending the possibility of antagonizing the oxidative stress generated in living organisms when the balance is in favor of free radicals as a result of the depletion of cell antioxidants.

Antioxidant activity of bisabolol: inhibitory effects on chemiluminescence of human neutrophil bursts and cell-free systems.

Human polymorphonuclear neutrophils (PMN), reactive oxygen species (ROS) and inflammatory reactions are closely interrelated, and increasing attention is being given to the search for new synthetic or natural antioxidant agents, capable of reducing ROS and consequent inflammation. It has been claimed that bisabolol (a monocyclic sesquiterpene alcohol) has an antioxidant/anti-inflammatory activity, but this has almost exclusively been investigated using chemical or biochemical tests. We studied the ability of bisabolol to interfere with ROS production (luminol-amplified chemiluminescence, LACL) during human PMN respiratory bursts induced by both corpusculate(Candida albicans)and soluble stimulants (N-formyl-methionyl-leucyl-phenylalanine, fMLP). LACL was also used to test cell-free systems (SIN-1 and H2O2/HOCl(-) systems) in order to investigate the presence of scavenging activity. After C. albicans stimulation, significant concentration-dependent LACL inhibition was observed at bisabolol concentrations ranging from 7.7 to 31 microg/ml; after the fMLP stimulus, significant LACL inhibition was observed at bisabolol concentrations ranging from 3.8 to 31 microg/ml. A similar effect was observed in the SIN-1 and H2O2/HOCl(-) systems. These findings draw the attention to the possible medical use of bisabolol as a means of improving the antioxidant network and restoring the redox balance by antagonising oxidative stress.

Visual evaluation of binding to mucosal cells of a medical device against the common cold.

The objective of this study was to investigate the possibility of visualizing the ability of hydroxypropylmethylcellulose (HPMC) and a nasal spray (First Defense), in which the bioadhesive is HPMC, to bind to human mucosal cells using inorganic (black carbon particles and Congo red dye) and organic markers (Escherichia coli). A significant reduction in the bacterial adhesiveness has been observed. Our findings indicate the possibility of counteracting the lock-and-key mechanism of microorganism adhesion using the bioadhesive properties of polymers, such as HPMC, in First Defense to prevent a possible contact between adhesins and complementary receptors.

Thymol inhibits Candida albicans biofilm formation and mature biofilm.

Candida albicans has a high propensity to develop biofilms that are resistant to traditional antifungal agents. Thymol is credited with a series of pharmacological properties including antimicrobial and antifungal effects. As C. albicans biofilms are known to be important factors underlying its virulence and pathogenicity, the aim of this study was to investigate whether thymol can interfere with biofilm formation as well as acting on mature biofilms. Tests of C. albicans strains ATCC 3153A and ATCC MYA 2876 showed that thymol interferes with the starting phases of biofilm production as well as with mature C. albicans biofilms. The metabolic activity of sessile cells was reduced by >90% at twice the minimum inhibitory concentration of planktonic cells. As biofilm is a multifactorial phenomenon, the multiple mechanisms of thymol (terpenes) could act on different steps in the evolution of mature biofilm.

Antioxidant effect of sulphurous thermal water on human neutrophil bursts: chemiluminescence evaluation.

BACKGROUND: The activities of the HS (sulfhydryl or thiolic) group in the cysteine of glutathione or various low-weight soluble molecules (thiolic drugs), such as N-acethylcysteine, mesna, thiopronine and dithiotreitol or stepronine and erdosteine (prodrugs), include its antioxidant activity in the airways during the release of reactive oxygen or nitrogen species (ROS, RNS) by polymorphonuclear neutrophils (PMNs) activated in response to exogenous or endogenous stimuli. OBJECTIVE: In addition to being administered by means of thiolic molecules, the HS group can also be given by means of the inhalation of sulphurous thermal water. The aim of this study was to investigate the effect of sulphurous thermal water on the release of ROS and RNS during the bursts of human PMNs. METHODS: The luminol-amplified chemiluminescence methodology was used to investigate the ROS and RNS released by PMNs stimulated with N-formyl-methionyl-leucyl-phenylalanine and phorbol-12-myristate-13-acetate, before and after incubation with sulphurous water. Effects on cell-free systems were also investigated. RESULTS: The water significantly reduced the luminol-amplified chemiluminescence of N-formyl-methionyl-leucyl-phenylalanine- andphorbol-12-myristate-13-acetate-activated PMNs on average from 0.94 to 15.5 mug/ml of HS, even after the addition of L-arginine, a nitric oxide (NO) donor. Similar findings have also been obtained in a cell-free system, thus confirming the importance of the presence of the HS group (reductive activity). CONCLUSIONS: The positive effects of the activity of sulphurous thermal waters has been partially based on the patients' subjective sense of wellbeing and partially on not always easy to quantify symptomatic (or general) clinical improvements. Our findings indicate that, in addition to their known mucolytic activity and trophic effects on respiratory mucosa, the HS groups present in the sulphurous thermal water of this spring also have antioxidant activity that contributes to the therapeutic effects of the water in upper and lower airway inflammatory diseases.

Effect of metabolite I of erdosteine on the release of human neutrophil elastase.

Erdosteine is a mucolytic drug whose metabolization gives rise to an active metabolite with an SH group (Met I), which reduces neutrophil release of reactive oxygen species and the peroxynitrite generated by the reaction of superoxide anion with nitric oxide, thus disrupting the phlogogenic loop sustained by activated neutrophils. Elastase, a serine proteinase released by activated human neutrophils, can degrade a wide variety of biomacromolecules including elastin and is considered to be pivotal in the pathogenesis of respiratory tract inflammation. The aim of this study was to examine whether, in addition to reducing the generation of free radicals, Met I can also interfere with the human neutrophil release of elastase. After the neutrophils were incubated with increasing amounts of Met I (1.25, 2.5, 5, 10, 20 microg/ml), elastase exocytosis was initiated by fMLP and measured using MeO-Suc-Ala-Ala-Pro-Val-MCA. The results showed that Met I significantly inhibits fMLP-induced elastase release by neutrophils in a concentration-dependent manner from 2.5 to 20 microg/ml. Our findings may be clinically relevant because the inhibitory effects were obtained at Met I concentrations that are achievable in the clinical setting. By reducing elastase and oxidant radical release (two major components of the inflammatory process), Met I have clinically useful anti-inflammatory effects.

Anti-inflammatory activity of thymol: inhibitory effect on the release of human neutrophil elastase.

Elastase, a serine proteinase released by activated human neutrophils, can degrade a wide variety of biomacromolecules including elastin, and is considered a marker of inflammatory diseases. As the logical strategy to protect tissue is to inhibit excessive elastase activity, experimental and clinical researches have concentrated on trying to find efficient elastase inhibitors. As thymol, one of the major components of thyme oil with a phenolic structure, has been credited with a series of pharmacological properties, that include antimicrobial and antioxidant effects, the aim of this study was to explore whether it can also interfere with the release of elastase by human neutrophils stimulated with the synthetic chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP). After the neutrophils were incubated with increasing amounts of thymol (2.5, 5, 10, 20 microg/ml), elastase release was initiated by fMLP and measured using MeO-Suc-Ala-Ala-Pro-Val-MCA. The results showed that thymol inhibited fMLP-induced elastase release in a concentration-dependent manner, with the effects of 10 and 20 microg/ml being statistically significant. The behavior of cytosolic calcium mobilization revealed by fura-2 closely resembled that of elastase, thus suggesting that they may be related. The hydrophobic nature of thymol means that it can approach ion channel proteins through the lipid phase of the membrane, alter the local environment of calcium channels and thus inhibit capacitative calcium entry. In brief, thymol inactivates calcium channels machinery, thus triggering a corresponding reduction in elastase. The antibacterial and antimycotic activity of thymol is already well known, but our findings that it inhibits elastase extend our knowledge of the anti-inflammatory activity of this interesting molecule that is already credited with antioxidant activity. These two latter characteristics make thymol a molecule that can have helpful effects in controlling the inflammatory processes present in many infections.

Antioxidant potential of thymol determined by chemiluminescence inhibition in human neutrophils and cell-free systems.

Thyme essential oil and thymol have antimicrobial, antifungal and antioxidant activities. Their antioxidant activity has been studied almost exclusively by means of chemical testing in order to be able to use it for food preservation purposes. The aim of this luminol amplified chemiluminescence (LACL) study was to investigate whether thymol can interfere with the production of reactive oxygen species, nitric oxide and the nitric oxide-derived peroxynitrite released by human neutrophils after activation by fMLP and PMA with and without the addition of the L-arginine (L-Arg) nitric oxide donor to the medium. The lowest thymol concentration that was still active in reducing LACL was 2.73 microg/ml, and there was a progressive linear inhibition of LACL from this concentration to 21.87 microg/ml, the highest thymol concentration investigated. This was also observed in the case of both fMLP and PMA stimulation with or without L-Arg. In cell-free systems using H(2)O(2)/HOCl(-) and SIN-1 as radical producers, a significant scavenging activity of thymol was present already at 0.08 and 0.68 microg/ml respectively, and these are very low concentrations. These findings can be related to the phenolic structure of thymol, because phenolic compounds have redox properties and play an important role in adsorbing and neutralizing free radicals and peroxynitrite, and in decomposing peroxides. Our findings in human neutrophils are pharmacologically relevant as they imply that thymol is a potential antioxidant and anti-inflammatory agent in human cells.