The Oxidation of Equol by Tyrosinase Produces a Unique Di-ortho-Quinone: Possible Implications for Melanocyte Toxicity

Equol (7-hydroxy-3-(4′-hydroxyphenyl)-chroman, EQ), one of the major intestinally derived metabolites of daidzein, the principal isoflavane found in soybeans and most soy foods, has recently attracted increased interest as a health-beneficial compound for estrogen-dependent diseases. However, based on its structure with two p-substituted phenols, this study aimed to examine whether EQ is a substrate for tyrosinase and whether it produces o-quinone metabolites that are highly cytotoxic to melanocyte. First, the tyrosinase-catalyzed oxidation of EQ was performed, which yielded three EQ-quinones. They were identified after being reduced to their corresponding catechols with NaBH₄ or L-ascorbic acid. The binding of the EQ-quinones to N-acetyl-L-cysteine (NAC), glutathione (GSH), and bovine serum albumin via their cysteine residues was then examined. NAC and GSH afforded two mono-adducts and one di-adduct, which were identified by NMR and MS analysis. It was also found that EQ was oxidized to EQ-di-quinone in cells expressing human tyrosinase. Finally, it was confirmed that the EQ-oligomer, the EQ oxidation product, exerted potent pro-oxidant activity by oxidizing GSH to the oxidized GSSG and concomitantly producing H₂O₂. These results suggest that EQ-quinones could be cytotoxic to melanocytes due to their binding to cellular proteins.     

Immunomodulatory effect of halophilic lactic acid bacterium Tetragenococcus halophilus Th221 from soy sauce moromi grown in high-salt medium

A halophilic lactic acid bacterium, Tetragenococcus halophilus, was found to possess an immunomodulatory activity that promotes T helper type 1 (Th1) immunity in addition to its important roles in soy sauce brewing. Strain Th221 was selected from 151 strains isolated from soy sauce (shoyu) moromi, since it induced strong interleukin (IL)-12 production by mouse peritoneal macrophages in vitro. The relationship between the salt concentration in the medium and the IL-12 production-inducing activity of this strain was investigated, and the activity was found to be strong when the bacteria were grown in medium containing > or =10% (w/v) salt. The Th1-promoting activity was also manifested in an in vivo mouse study, since Th1-dependant contact sensitivity was augmented and Th2 immunity, as evaluated by specific immunoglobulin E production, was suppressed following oral ingestion of Th221. Based on these findings, Th221 administration may be useful for improving allergic symptoms.     

Analgesic Mechanisms of Steroid Ointment against Oral Ulcerative Mucositis in a Rat Model

Despite the long history of use of steroid ointments for oral mucositis, the analgesic mechanism has not been fully elucidated. In this study, we examined the effects of triamcinolone acetonide (Tmc) on oral ulcerative mucositis-induced pain in conscious rats by our proprietary assay system. Based on evaluations of the physical properties and retention periods in the oral mucosa of human volunteers and rats, we selected TRAFUL^® ointment as a long-lasting base. In oral ulcerative mucositis model rats, TRAFUL^® with Tmc suppressed cyclooxygenase-dependent inflammatory responses with upregulations of glucocorticoid receptor-induced anti-inflammatory genes and inhibited spontaneous nociceptive behavior. When an ointment with a shorter residual period was used, the effects of Tmc were not elicited or were induced to a lesser extent. Importantly, TRAFUL^® with Tmc also improved oral ulcerative mucositis-induced mechanical allodynia, which has been reported to be independent of cyclooxygenase. Ca²⁺ imaging in dissociated trigeminal ganglion neurons showed that long-term preincubation with Tmc inhibited the hypertonic stimulation-induced Ca²⁺ response. These results suggest that the representative steroid Tmc suppresses oral ulcerative mucositis-induced pain by general anti-inflammatory actions and inhibits mechanical sensitivity in peripheral nerves. For drug delivery, long-lasting ointments such as TRAFUL^® are needed to sufficiently induce the therapeutic effects.     

Visual selection from multielement displays: Measuring and modeling effects of exposure duration.

In a partial-report experiment, subjects reported the digits from a circular array of digits and letters terminated by a pattern mask. Individual frequency distributions of the number of correctly reported digits were analyzed as functions of number of digits (2, 4, or 6) and number of letters (0, 2, 4, 6, or 8) at nine exposure durations ranging from 10 to 200 ms. The distributions (hundreds of data points per subject) were accurately predicted by a four-parameter fixed capacity independent race model that assumes exponentially distributed processing times, limitations in both processing capacity and storage capacity, and time-invariant selectivity. Estimated from the data, processing capacity C was 45 items/s, selectivity α (ratio between the amount of processing capacity devoted to a distractor and the amount devoted to a target) was 0.48, short-term storage capacity K was 3.5 items, and the longest ineffective exposure duration t? was 18 ms. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Binding and catalytic properties of Xenopus (6-4) photolyase

Xenopus (6-4) photolyase binds with high affinity to DNA bearing a (6-4) photoproduct and repairs it in a light-dependent reaction. To clarify its repair mechanism of (6-4) photolyase, we determined its binding and catalytic properties using synthetic DNA substrate which carries a photoproduct at a single location. The (6-4) photolyase binds to T[6-4]T in double-stranded DNA with high affinity (KD = 10(-9)) and to T[6-4]T in single-stranded DNA and T[Dewar]T in double- and single-stranded DNA although with slightly lower affinity (KD = approximately 2 x 10(-8)). Majority of the T[6-4]T-(6-4) photolyase complex dissociates very slowly (koff = 2.9 x 10(-5) s-1). Its absolute action spectrum without a second chromophore in the 350-600 nm region closely matches the absorption spectrum of the enzyme. The quantum yield (phi) of repair is approximately 0.11. The fully reduced form (E-FADH-) of (6-4) photolyase is catalytically active. Direct analysis of the photoreactivated product showed that (6-4) photolyase restores the original pyrimidines. These findings demonstrate that cis, syn-cyclobutane pyrimidine dimer photolyase and (6-4) photolyase are quite similar, but they are different with regard to the binding properties.     

Techniques for Supporting Dynamic and Adaptive Workflow

The unpredictability of business processes requires that workflowsystems support exception handling with the ability to dynamicallyadapt to the changing environment. Traditional approaches to handlingthis problem have fallen short, providing little support for change,particularly once the process has begun execution. Further,exceptions vary widely in their character and significance,challenging the application of any single approach to handling them.We briefly discuss the classification of exceptions, highlightingdiffering impacts on the workflow model. Based on this discussion, wesuggest principal goals to address in the development of adaptiveworkflow support, including strategies for avoiding exceptions,detecting them when they occur, and handling them at various levels ofimpact. We then identify a number of specific approaches to supportingthese goals within the design of a workflow system infrastructure.Finally, we describe the implementation of many of these approaches inthe Endeavors workflow support system.     

Parts-mix analysis in recycling-oriented manufacturing systems for multiple-item production

One solution to environmental problems is to construct a recycling-oriented manufacturing system, in which waste products after consumption are collected, and reusable parts in the collected products are employed to produce new products. A parts-mix in the manufacturing system for multiple-item production is analysed by taking into consideration the merits and demerits of the usage of common parts over multiple products and special parts utilized for individual products. The parts-mix problem is formulated to minimize the life-cycle cost, which consists of production costs and recycling costs. The property of the optimal parts-mix is clarified, and a decision procedure to solve the parts-mix is developed. The procedure decides whether the common parts are effective or not.     

Effect of temperature on the stability of various peptidases during peptide-enriched soy sauce fermentation

We previously developed a peptide-enriched soy sauce-like seasoning called Fermented Soybean Seasoning (FSS) with high-temperature fermentation, and we have reported the antihypertensive effects of FSS. Seryl-tyrosine (Ser-Tyr) and glycyl-tyrosine (Gly-Tyr) were identified from FSS as active constituents in the antihypertensive effects. They were found to be particularly enriched in FSS; more so than in regular soy sauce. In the present study, we clarified one of the mechanisms underlying the accumulation of these bioactive peptides during high temperature soy sauce brewing. Crude enzyme extracts were prepared from model soy sauce mash (moromi) fermented at various temperatures. Leucine aminopeptidase-I, II, and seryl-tyrosine hydrolytic activity were found to decrease in the moromi incubated at the fermentation temperature of FSS whereas almost no decrease was observed in that of regular soy sauce. The concentrations of ACE inhibitory peptides, Ser-Tyr and Gly-Tyr, in the moromi incubated at high temperature were revealed to be higher than those at low temperature through quantitative LC-MS/MS analysis. These results suggested that the peptidases responsible for degrading low molecular weight bioactive peptides were inactivated during the high temperature fermentation, thus, these peptides would be likely to remain in the high temperature fermentation.