Inhibition of tyrosinase by green tea components.

The pigment melanin in human skin is a major defense mechanism against ultraviolet light of the sun, but darkened skin color, which is the result of increased and redistributed epidermal melanin, could be a serious aesthetic problem. Epidemiologically, it is well known that the consumption of green tea may help prevent cancers in humans and also reduce several free radicals including peroxynitrite. In the present study, to assess the efficacy of the inhibition of mushroom tyrosinase (monophenol monooxygenase EC 1.14.18.1), ten kinds of Korean traditional teas were screened for their tyrosinase inhibitory activity. Green tea was the strongest inhibitor, and the major active constituents in the tea are (-)-epicatechin 3-O-gallate (ECG), (-)-gallocatechin 3-O-gallate (GCG), and (-)-epigallocatechin 3-O-gallate (EGCG). All are catechins with gallic acid group as an active site. The kinetic analysis for inhibition of tyrosinase revealed a competitive nature of GCG with this enzyme for the L-tyrosine binding at the active site of tyrosinase.

Dietary modulation of prostanoid synthesis in the aging process: role of cyclooxygenase-2.

To investigate the role of cyclooxygenase (COX) as a mediator for prostaglandin synthesis in relation to generation of reactive oxidant species (ROS), we quantitated the COX-derived ROS generation and the gene expression of COX-2, an inducible form of COX in aged kidney. In addition, the modulation by dietary restriction was investigated. COX-derived ROS generation increased with age in ad libitum (AL) rats, but dietary restriction (DR) suppressed the level. The amounts of COX-2 protein and mRNA increased with age in AL rats but maintained at low levels in DR group. It was found that the binding characteristics of a nuclear transcription factor, NF-kappaB were altered by aging. The binding activity of NF-kappaB in aged kidney was significantly enhanced with the corresponding increase in mRNA and protein levels. These increases were closely in parallel to the increased ROS generation and gene expression of COX-2. The COX activity shown by NF-kappaB activation and the ROS generation by COX-mediated process were all modulated by DR. Our results suggest that the upregulation of COX-2 during aging may play an important role in many age-related diseases associated with aging process. And this upregulation was attenuated by DR. We propose that the modulation of the redox-sensitive transcription factor may well be a part of the mechanisms underpinning the anti-oxidative action of DR.

Xanthine dehydrogenase/xanthine oxidase and oxidative stress.

Xanthine dehydrogenase (XDH) and xanthine oxidase (XOD) are single-gene products that exist in separate but interconvertible forms. XOD utilizes hypoxanthine or xanthine as a substrate and O2 as a cofactor to produce superoxide (·O2 (-)) and uric acid. XDH acts on these same substrates but utilizes NAD as a cofactor to produce NADH instead of ·O2 (-) and uric acid. XOD has been proposed as a source of oxygen radicals in polymorphonuclear, endothelial, epithelial, and connective tissue cells. However, several questions remain about the physiological significance and functions of XOD on aging and oxidative stress. XOD is reported to play an important role in cellular oxidative status, detoxification of aldehydes, oxidative injury in ischemia-reperfusion, and neutrophil mediation. For example, XOD may serve as a messenger or mediator in the activation of neutrophil, T cell, cytokines, or transcription in defense mechanisms rather than as a free radical generator of tissue damage. Emerging evidence on the synergistic interactions of ·O2 (-), a toxic product of XOD and nitric oxide, may be another illustration of XOD involvement in tissue injury and cytotoxicity in an emergent condition such as ischemia or inflammation.

An ankyrin-related gene (unc-44) is necessary for proper axonal guidance in Caenorhabditis elegans.

Caenorhabditis elegans unc-44 mutations result in aberrant axon guidance and fasciculation with inappropriate partners. The unc-44 gene was cloned by transposon tagging, and verified by genetic and molecular analyses of six transposon-induced alleles and their revertants. Nucleotide sequence analyses demonstrated that unc-44 encodes a series of putative ankyrin-related proteins, including AO49 ankyrin (1815 aa, 198.8 kD), AO66 ankyrin (1867 aa, 204 kD), and AO13 ankyrin (< or = 4700 aa, < or = 517 kD). In addition to the major set of approximately 6 kb alternatively spliced transcripts, minor transcripts were observed at approximately 3, 5, 7, and 14 kb. Evidence is provided that mutations in the approximately 14-kb AO13 ankyrin transcript are responsible for the neuronal defects. These molecular studies provide the first evidence that ankyrin-related molecules are required for axonal guidance.