Biomarkers of oxidative/nitrosative stress: an approach to disease prevention.

Oxidative/nitrosative stress is responsible for a variety of degenerative processes in some human diseases. Measurement of oxidatively/nitrosatively modified DNA, proteins, lipids, and sugars in biological samples has been expected to detect appropriate biomarkers for diseases in which reactive oxygen/nitrogen species are involved. Recently, the application of these biomarkers to epidemiological studies has resulted in a new discipline, molecular epidemiology, which provides the opportunity for better understanding of their causal relation with disease outcomes in a population level. In this brief review, we cover some specific biomarkers of oxidative/nitrosative stress with regard to the commonly used analytical methods for these biomarkers, their integration with epidemiology, and their application in antioxidant intervention trials, with an emphasis on those applicable to human studies and their potentialities for disease prevention

Mammalian Acatalasemia: The Perspectives of Bioinformatics and Genetic Toxicology

The molecular defects in the catalase gene, levels of m-RNA and properties of the residual catalase studied by scientists are reviewed in human (Japanese, Swiss and Hungarian) and non-human (mouse and beagle dog) acatalasemia with reference to the bioinformatics. Japanese acatalasemia-I, the G to A transition at the fifth position of intron 4 of the catalase gene, limited the correct splicing of the mRNA and synthesized trace catalase with normal properties. Hungarian acatalasemia type C showed a splicing mutation. In the Japanese acatalasemia II and the type A and B of Hungarian acatalasemia, the deletion or insertion of nucleotides was observed in the coding regions, and the frame shift altered downstream amino acid sequences and formed truncated proteins. In the Hungarian acatalasemia D, the substitution of a nucleotide in the exon was found. In mouse and beagle dog acatalasemia, the substitution of nucleotides in the coding regions was also observed. Studies of residual catalase in Swiss, mouse and beagle dog acatalasemia showed that aberrant catalase protein degrades more quickly than normal catalase in cells. The experimental research in genetic toxicology concerning the effect of oxidative stressors (nitrogen monoxide, nitrogen dioxide and so on) on Japanese acatalasemic blood and acatalasemic mice is described. The clinical features of Japanese and Hungarian acatalasemic subjects are also described.</p

Relation of Prenatal and Postnatal Status to Calcaneus Quantitative Ultrasound in Adolescents

This study aimed to elucidate the relationship of prenatal and/or postnatal factors, including acquired factors, with the calcaneus stiffness index as measured by quantitative ultrasound (QUS-SI) in adolescents. We recruited 1,143 adolescents with a mean age of 14.8±1.8 years (501 boys and 642 girls). The subjectsʼ calcaneus QUS-SI was measured using an ultrasound bone densitometer. We also measured the subjectsʼ height, weight, and grip strength. Data on prenatal and postnatal factors were obtained from maternal and child health handbooks. A self-reporting questionnaire was used to obtain information on subjectsʼ secondary sexual characteristics and lifestyle factors. We found that maternal weight gain during pregnancy was independently associated with calcaneus QUS-SI in girls, and that grip strength was also significantly associated with calcaneus QUS-SI in both sexes. The present findings suggest that excessive restriction of maternal weight gain would have a negative effect on the calcaneus QUS-SI of girls, and that exercise and strength-building activities are likely to result in a higher calcaneus QUS-SI in both sexes of adolescents

Involvement of Xanthine Oxidoreductase-related Oxidative Stress in a Dermatophagoides farinae-induced Asthma Model of NC/Nga Mice

Oxidative stress is widely known to play a role in asthma. However, the contribution of xanthine oxidoreductase (XOR) as a source of the superoxide anion radical (O2－) in oxidative stress associated with asthma has not yet been examined in detail. Here we investigated pathophysiological changes in XOR in an experimental model of asthma induced by the house dust mite Dermatophagoides farinae (Df). In the lungs of Df-treated mice, the production of O2－ from XOR increased and the nitrite concentrations decreased, whereas the protein expression of XOR remained unchanged. Moreover, the protein expression levels of XOR and the hydrogen peroxide (H2O2) concentrations in bronchoalveolar lavage fluid (BALF) were higher in the Df-treated mice than in saline-treated mice. Immunohistochemically, although XOR was highly localized in the bronchial epithelial cells of the saline-treated mice, immunostaining for XOR was absent in the bronchial epithelium of Df-treated mice. These results suggest that oxidative stress is up-regulated by increases in the conversion of the dehydrogenase form (xanthine dehydrogenase; XDH) of XOR to the oxidase form (xanthine oxidase; XOD)

重金属による食細胞フリーラジカル産生刺激と細胞内情報伝達機構

金沢大学医学部酸化亜鉛(ZnO)フュームによる金属熱の発症機序を解明する目的で、ラット腹腔内浸出性好中球やマウスマクロファージ様細胞株Raw264.7などの貪食細胞を用いて、ZnOの活性化作用とその機序を検討した.ZnOは、グルタチオン(GSH)やチオール化合物の添加により活性化作用が増強されることがわかった。ZnO単独では、好中球やマクロファージの細胞膜のCR3受容体に認識され、GSHによりFcγRやその他の受容体を介して、GTP結合蛋白、カルモジュリン、プロテインキナーゼC、チロシンキナーゼ、細胞内カルシウムなどの細胞内情報伝達機構の関与のもとにNADPHオキシダーゼの活性化によりO_2さらにH_2O_2が増加することがわっかた。ZnOとZnO+GSH刺激による好中球活性化作用の違いを検討すると、ZnOは細胞膜のNADPHオキシダーゼ以外に細胞質の一酸化窒素合成酵素(NOS)からO_2産生を刺激しパーオキシナイトライト(ONOO)を発生させるのに対し、ZnO十GSHは、ミトコンドリアに局在するといわれるモノアミンオキシダーゼ(MAO)を活性化しH_2O_2の産生を増強している可能性が示唆された。このことは、核転写因子の活性化やアポトーシスの発生機序に関与する新しいH_2O_2の発生源としての可能性を有しているだけではなく、MAOと細胞内情報伝達機構の関係を繋ぐ新しい発見の可能性がある。さらに、Raw246.7を用いた実験から、GSHはZnO粒子の貪食作用を増強させることが認められた。本研究より、亜鉛熱は、ZnO粒子に内因性のGSHが作用することによって発症する可能性が示唆された。The activation mechanisms of intraperitoneal-eluted rat neutrophils and mouse macrophage -like cell line Raw 264.7 by zinc oxide (ZnO) was investigated to explore the etiological mechanisms of metal fume fever induced by ZnO.The activation of phagocytes by ZnO was enhanced by the addition of glutathione (GSH) and thiol compounds. Superoxide (O2^-) and hydrogen peroxide (H2O2) may be produced by NADPH oxidase via the intracellular signal transduction pathway such as GTP-binding protein, protein kinase C, tyrosine kinase, and intracellar calcium and membraneous CR3 if ZnO was added as a stimulant to neutrophils. If GSH was added to ZnO, the production of acitive oxygen species was dependent on FcgammaR in addition to CR3. The differnce in the activation of neutrophils by ZnO and ZnO plus GSH was investigated. The activation of neutrophils by ZnO may be dependent on the production of O2^- and peoxynitrite (ONOO^-) via NADPH oxidase and nitric oxide synthase (NOS). The activation of neutrophils by ZnO plus GSH may participated in the elevated H2O2 via the activation of monoamine oxidase (MAO) mainly localized in mitochodrium. These results suggest pot only the new evidence of the origin of H2O2 associated with the activation of nuclear transcription factor and the etiology of apoptosis but also the discovery of the relationship between MAO and intracellar signal transduction mechanisms. Moreover, from the results of Raw 246.7, GSH promoted the cleaning of ZnO particles by phagocytosis.From this research project, it is speculated that zinc fume fever may be caused by the intrinsic factor GSH in addition to ZnO particles.研究課題/領域番号:08457114, 研究期間(年度):1996 – 1998出典：研究課題「重金属による食細胞フリーラジカル産生刺激と細胞内情報伝達機構」課題番号08457114（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-08457114/084571141998kenkyu_seika_hokoku_gaiyo/）を加工して作

ナノテクノロジー粒子状化学物質とアレルギー発症に関する研究

金沢大学医薬保健研究域医学系ナノテクノロジー仕様の酸化チタン及び酸化亜鉛をそれぞれ、8μg/50μlをNC/Ngaマウスに5日間連続点鼻し、さらに11日目にチャレンジ投与した喘息発症実験で、12日目に肺の組織学的検討と供に肺胞洗浄液の細胞分画を観察した。また、我々がこのマウスを用いて、ダニタンパク質抽出液Df抗原を点鼻し作成した、喘息モデルに対して、酸化チタン40μg/50μlを11日目のDfチャレンジ投与の直前に点鼻投与し、肺胞洗浄液中の細胞分画への影響を検討した。その結果、酸化チタンは肺胞マクロファージに取り込まれ一部気管支周囲に炎症細胞の浸潤があり、NO合成酵素の一つであるiNOSが一部誘導されていたが、酸化亜鉛にはそのような作用は認められなかった。また、酸化亜鉛、酸化チタンによる肺胞洗浄液細胞分画は、ほとんどがマクロファージであり好酸球の増加は認められなかった。Df点鼻投与により発症した喘息モデルでは、肺胞洗浄液中には、多数のマクロファージ、好中球、好酸球の浸出が認められたのに対して、酸化チタンをチャレンジ前投与した場合、マクロファージ、好酸球の浸出を抑制した。以上より、酸化チタンには、吸入毒性がある可能性があるが、易アレルギー性は少ないと思われた。しかしながら、酸化チタンにはiNOS誘導のようなTh1刺激作用があり、好酸球の浸潤を抑制する作用があることから、喘息を抑制する可能性がある。治療面への応用を考慮し、この後、さらに検討する必要があると思われる。研究課題/領域番号:17659174, 研究期間(年度):2005 – 2006出典：「ナノテクノロジー粒子状化学物質とアレルギー発症に関する研究」研究成果報告書　課題番号17659174（KAKEN：科学研究費助成事業データベース（国立情報学研究所））（https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-17659174/)を加工して作

フリーラジカル産生からみた重金属による白血球活性化の分子機構

金沢大学医学部金属熱の発症機序を解明する目的で亜鉛を中心とした重金属、さらに金属キレート剤による肺胞マクロファージ及び好中球の活性化機序をスーパーオキシド産生を指標として細胞-内情報伝達機構の面から解析した。これらの貧食細胞からのスーパーオキシド産生刺激能は硫酸亜鉛(ZnSO4)<酸化亜鉛(ZnO)<水酸化亜鉛(Zn(OH2))の順に増強した。これら亜鉛及び亜鉛酸化物を認識する貧食細胞の細胞内情報伝達機構の違いとして次のことがわかった。亜鉛イオンに対しては、カルシュウム非依存性でプロテインキナーゼCの関与が少なく、スーパーオキシド由来ではない過酸化水素の発生が認められ、未知の細胞内情報伝達経路の関与が考えられた。酸化亜鉛に対しては、細胞外カルシウム非依存性で還元型グルタチオン(GSH)にプライミング減少による刺激作用の増強がみられ、チロシンキナーゼ型の受容体を介している可能性が示唆された。水酸化亜鉛に対しては、細胞外カルシウム依存性で細胞内への流入がみられプロテインキナーゼCの活性化が関与しており、粒子の貧食が主な機構であると思われた。これらのことより、亜鉛化合物の微妙な分子構造の違いを貧食細胞が認識できることがわかった。水酸化亜鉛による好中球の活性化とNO(一酸化窒素)の関係では、産生されるNOを著名に減少させる作用を有しているが、これはスーパーオキシドとの反応によるものか又は直接吸着されことが考えられた。さらに白血球接着因子であるLAF-1は水酸化亜鉛による活性化には何ら関与を示さなかった。その他、カドミウムと金属キレート剤エチルジチオカ-バメートに好中球活性化作用が認められ、ガドリニウムに活性化貧食細胞の抑制作用等も認められた。The mechanisms of activation of alveolar macrophages and neutrophils induced by heavy metals, mainly zinc, and then metal chelator were investigated by monitoring the release of superoxide in order to resolve the pathogenesis of metal fume fever from a view point of signal transduction. The effciency of superoxide radical production by zinc compounds was ranked in order as zinc sulfate (ZnSO4) <<zinc oxide (ZnO) <zinc hydroxide (Zn(OH)2). The mechanisms of the signal transduction of three zinc compounds varied with each other. Zinc ions from zinc sulfate stimulated hydrogen peroxide. The hydrogen peroxide production was not superoxide, independent on calcium and less dependent on protein kinase C.It was suggested that the activation of neutrophils by zinc ions might be dependent on unknown signal transduction pathway. Zinc oxide stimulated neutrophils in the absence of extracellular calcium and the stimulation was enhanced by addition of GSH (a priming effect). Therefore, it was suggested that zinc oxide might stimulate superoxide production of neutrophils via receptors closely related with tyrosine protein kinase. On the other hand, zinc hydroxide stimulated neutrophils by the process of the influx of calcium from extracellular fluids and the activation of protein kinase C.The major mechanism of neutrophils activation by zinc hydroxide may be dependent on the phagocytosis. These results suggested that a subtle molecular change of zinc compounds might be contributed to the recognition of the phagocytes.Stimulation of neutrophils by zinc hydroxide reduced NO release from the same cells. This phenomenon may result from the interaction with NO and superoxide or NO and zunc hydroxide. There was no expression of LAF-1 on the surface of cell membrane with the stimulation of zinc hydroxide. A monolonal antibody to LAF-1 did not suppress the activation of neutrophils by zinc hydroxide.The activation of neutrophils by cadmium and a metal chelator of diethydithiocarbamate, and the suppression of activated phagocytes by gadolinium were also observed.研究課題/領域番号:05454228, 研究期間(年度):1993 – 1995出典：研究課題「フリーラジカル産生からみた重金属による白血球活性化の分子機構」課題番号05454228（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （https://kaken.nii.ac.jp/ja/report/KAKENHI-PROJECT-05454228/054542281995kenkyu_seika_hokoku_gaiyo/）を加工して作

The Relationship between Joint Pain and Climate Conditions in Japan

This study aimed to determine whether there was any association between the regional climate and the proportion of people with joint pain. Regional climate data between 1971 and 2000 were obtained from the Japan Meteorological Agency. The variables used in the cluster analysis included sunlight hours, amount of precipitation, number of days with precipitation, and temperature. The regional proportion of people with joint pain was obtained from the National Survey for Health in 2001. After performing a cluster analysis, one-way ANOVA and Welch's test were used to determine whether the climate characteristics of the clusters were significantly different. Within each cluster, stepwise multiple linear regression analyses were performed. We found that sunlight hours showed a direct, negative association with the proportion of people with joint pain (adjusted R2＝0.532, p＝0.016) in cluster 1, which was characterized as the region with the fewest total hours of sunlight, less precipitation, a modest number of rainy days, and low temperature. In the other clusters, the regional female population rate (cluster 2) and the senior population rate (cluster 3, 4) were the primary predictors. We concluded that the degree of exposure to sunlight may play a crucial role in prevention of joint pain. This finding should encourage people to set aside some time for staying outdoors in their daily lives

Relationship of psychological factors with physical activity stage of change in prime-and middle-aged Japanese

The present study aimed to investigate the status of physical activity and the differences in psychological factors associated with physical activity from the perspective of transtheoretical model stages between prime- and middle-aged Japanese. The study involved 375 prime-aged volunteers (175 men, 200 women) and 557 middle-aged volunteers (247 men, 310 women) living in Kuse, a town in Okayama Prefecture, Japan. We found that the prime-aged men at the preparation stage had significantly higher self-efficacy scores than at the contemplation stage (p&#60;0.01). Middle-aged men had significantly higher self-efficacy scores at the contemplation stage than at the precontemplation stage (p&#60;0.001). Middle-aged women, meanwhile, had significantly higher self-efficacy scores at the maintenance stage than at the action stage (p&#60;0.01), and at the contemplation stage than at the precontemplation stage (p&#60;0.001). The present findings provide valuable information about the differences in psychological factors affecting physical activity between prime-aged and middle-aged community-dwelling Japanese. This information may be useful to health professionals as they develop effective community-based intervention programs for target populations.</p

Effects of mask fitness and worker education on the prevention of occupational dust exposure

To decrease the incidence of pneumoconiosis, we examined dust protective mask performance and its relation to pulmonary function as well as the effects of worker education on the proper wearing of masks. One hundred and seventy-eight workers from 15 factories subject to dust exposure participated in this study. All participants were interviewed to obtain relevant personal information and underwent both a mask leakage and a pulmonary function test. The mask leakage was expressed as a percentage, with under 10% leakage indicating that the dust protective mask worked efficiently. In addition, 23 workers from 2 factories were educated on how to wear masks properly. The average mask leakage was 24.3%, and 58% of workers wore ineffective masks. Though pulmonary function was almost normal, the percent vital capacity (%VC) tended to be lower depending on the mask leakage. Mask education, which was very easy and took only a short time, dramatically decreased average mask leakage from 32.1% to 10.5% (p0.001). Educating workers to wear masks properly might prevent the worsening of pulmonary function in response to dust exposure. Appropriate mask fitness by education could be useful in preventing the development of pneumoconiosis.</p