Evaluation of radical scavenging properties of shikonin.

With the aim of developing effective anti-inflammatory drugs, we have been investigating the biochemical effects of shikonin of "Shikon" roots, which is a naphthoquinone with anti-inflammatory and antioxidative properties. Shikonin scavenged reactive oxygen species like hydroxyl radical, superoxide anion (O2 (•-)) and singlet oxygen in previous studies, but its reactivity with reactive oxygen species is not completely understood, and comparison with standard antioxidants is lacking. This study aimed elucidation of the reactivity of shikonin with nitric oxide radical and reactive oxygen species such as alkyl-oxy radical and O2 (•-). By using electron paramagnetic resonance spectrometry, shikonin was found unable of reacting with nitric oxide radical in a competition assay with oxyhemoglobin. However, shikonin scavenged alkyl-oxy radical from 2,2'-azobis(2-aminopropane) dihydrochloride with oxygen radical absorbance capacity, ORAC of 0.25 relative to Trolox, and showed a strong O2 (•-)-scavenging ability (42-fold of Trolox; estimated reaction rate constant: 1.7 × 10(5) M(-1)s(-1)) in electron paramagnetic resonance assays with CYPMPO as spin trap. Concerning another source of O2 (•-), the phagocyte NADPH oxidase (Nox2), shikonin inhibited the Nox2 activity by impairing catalysis when added before enzyme activation (IC50: 1.1 µM; NADPH oxidation assay). However, shikonin did not affect the preactivated Nox2 activity, although having potential to scavenge produced O2 (•-). In conclusion, shikonin scavenged O2 (•-) and alkyl-oxy radical, but not nitric oxide radical.

2,2'-Azobis(isobutyronitrile)-derived alkylperoxyl radical scavenging activity assay of hydrophilic antioxidants by employing EPR spin trap method.

As interest in the study of antioxidant intake from foods and other agricultural products increases, methods for performing radical scavenging activity assays based on the electron paramagnetic resonance spectroscopic method, in which there is no interference from the sample color and turbidity, are required. In this study, we have developed a rapid and simple electron paramagnetic resonance based assay to evaluate the alkylperoxyl radical scavenging activity of several antioxidants. The alkylperoxyl radical species was generated by the photolysis of azo-radical initiator 2,2'-azobis(isobutyronitrile), in which the radical generation rate and period were controlled by the illumination light. The relative alkylperoxyl radical scavenging activity was obtained by a simple formula of competing reaction of antioxidant and spin trap toward the oxygen radical. The scavenging activities toward alkylperoxyl radical and alkoxy radical species were evaluated in six antioxidants. Although quercetin showed the highest activity toward both radicals, the order of the relative activities in the other antioxidants was different mutually between the alkylperoxyl radical and the alkoxyl radical. This alkylperoxyl radical scavenging activity assay based on electron paramagnetic resonance spectroscopy is useful for evaluation of colored and turbid food samples.

Recognition of human emotions from cerebral blood flow changes in the frontal region: a study with event-related near-infrared spectroscopy.

BACKGROUNDS AND PURPOSE: The aim of this study is to develop a near-infrared spectroscopy (NIRS)-based system that recognizes pleasant and unpleasant human emotions based on cerebral blood flow (CBF) in order to understand the minds of patients whose brain function is severely impaired. The forehead region is easily accessible to NIRS measurements, whereas the role of the anterior prefrontal cortex (PFC) in the processing of emotion remains to be elucidated. METHODS: Initially, using event-related NIRS we examined changes in oxygenated hemoglobin (oxy-Hb) as an indicator of regional CBF changes, which reflect brain activity directly related to emotions, but not to cognitive operations in the anterior frontal regions, during viewing affective pictures. The event-related potentials (ERPs), systemic blood pressure, and pulse rate were also measured simultaneously. RESULTS: The event-related analysis of changes in oxy-Hb for a 6 s-picture presentation period showed that very unpleasant emotion was accompanied by an increase in oxy-Hb in the bilateral ventrolateral PFCs, while very pleasant emotion was accompanied by a decrease in oxy-Hb in the left dorsolateral PFC. There were no significant differences in either ERPs or autonomic nervous system activities between the two emotional states. CONCLUSION: These findings suggest the possibility of recognizing patients' emotions from CBF changes.

An oxygen radical absorbance capacity-like assay that directly quantifies the antioxidant's scavenging capacity against AAPH-derived free radicals.

A new method is proposed for the evaluation of oxygen radical absorbance capacity (ORAC). The current fluorescence-based ORAC assay (ORAC-FL) is an indirect method that monitors the antioxidant's ability to protect the fluorescent probe from free radical-mediated damage, and an azo-radical initiator, AAPH (2,2-azobis(2-amidinopropane) dihydrochloride), has been used as a thermal free radical source. The new ORAC assay employs a short in situ photolysis of AAPH to generate free radicals. The electron paramagnetic resonance (EPR) spin trapping method was employed to identify and quantify AAPH radicals. In the presence of antioxidant, the level of AAPH radicals was decreased, and ORAC-EPR values were calculated following a simple kinetic formulation. Alkyl-oxy radical was identified as the sole decomposition product from AAPH; therefore, we concluded that ORAC-FL is the assay equivalent to alkyl-oxy radical scavenging capacity measurement. ORAC-EPR results for several antioxidants and human serum indicated that the overall tendency is in agreement with ORAC-FL, but absolute values showed significant discrepancies. ORAC-EPR is a rapid and simple method that is especially suitable for thermally labile biological specimens because the sample heating is not required for free radical production.

Quantitative evaluation of the relative contribution ratio of cerebral tissue to near-infrared signals in the adult human head: a preliminary study.

Combining spatially- and time-resolved spectroscopies. we attempted to quantitatively evaluate the contribution ratio of the partial mean pathlength of cerebral tissue to the observed overall mean pathlength, in which haemoglobin concentrations were selectively changed by administration of acetazolamide. When acetazolamide was administered, the observed increases in oxygenated haemoglobin depended on the probe distance, which became progressively larger at distances of 2, 3 and 4 cm. Increases in oxygen saturation were detected at 3 and 4 cm spacing, but not at 2 cm. Assuming that the modified Lambert-Beer's law can exist in the inhomogeneous structure of the head, then, we could estimate the contribution ratio of the cerebral tissue to optical signals at the probe distances of 2, 3 and 4 cm as 33%, 55% and 69%, respectively. Using these values, we recalculated acetazolamide-induced concentration changes in oxygenated-haemoglobin in the cerebral tissue, which resulted in the same values at distances of 2, 3 and 4 cm as expected. Thus, our present method opened the door to the possibility of selectively obtaining optical signals attributed to cerebral tissue.