Preconditioning with subneurotoxic allyl nitrile: Protection against allyl nitrile neurotoxicity

金沢大学医薬保健研究域医学系High-dose cruciferous allyl nitrile can induce behavioral abnormalities in rodents, while repeated exposure to allyl nitrile at subneurotoxic levels can increase phase 2 detoxification enzymes in many tissues, although the brain has not been investigated yet. In the present study, we examined the effect of 5 days repeated exposure to subneurotoxic allyl nitrile (0-400 μmol/kg/day) on the brain. Elevated glutathione S-transferase activity was recorded in the striatum, hippocampus, medulla oblongata plus pons, and cortex. Enhancement of quinone reductase activity was observed in the medulla oblongata plus pons, hippocampus, and cortex. In the medulla oblongata plus pons, elevated glutathione levels were recorded. Following repeated subneurotoxic allyl nitrile exposure (0-400 μmol/kg/day), mice were administered a high-dose allyl nitrile (1.2 mmol/kg) which alone led to appearance of behavioral abnormalities. Compared with the 0 μmol/kg/day group, animals in the 200 and 400 μmol/kg/day pre-treatment groups exhibited decreased behavioral abnormalities and elevated GABA-positive cell counts in the substantia nigra pars reticulata and the interpeduncular nucleus. These data suggest that repeated exposure to subneurotoxic levels of allyl nitrile can induce phase 2 enzymes in the brain, which together with induction in other tissues, may contribute to protection against allyl nitrile neurotoxicity. © 2009 Elsevier Ltd. All rights reserved

転写因子・作用蛋白の同定による表皮の分化・脱核機構の解明

金沢大学医薬保健研究域医学系分化誘導を行う/行ないkeratinocyte培養細胞間で差の認められるmRNAを同定し、 分化誘導の違いのあるeratinocyte培養細胞間で差の認められるmRNAを同定する。同遺伝子発現蛋白は骨芽細胞分化の責任遺伝子と考えられ、培養骨芽細胞に転写因子・作業蛋白のmRNAsを導入、knock downすることで生じる機能変化を同定する。皮膚3次元モデルから、表皮本来各層; 基底層、有棘層、顆粒層、角質層から分離・摘出したmRNAs解析し、遺伝子の意義を特定する。細胞での機能、遺伝子導入による転写因子・作業蛋白の活性化・不活性化による表皮分化促進・抑制で細胞間での認識の違いも明らかにできる。長さの異なるKLKs・SPINK5のプロモーターおよびGATA3のプロモーターをPCR増幅し、ルシフェレースレポーターシステムに組み込んだクローンを作成した。最長 2000baseから最短100baseまで、各10クローンを得ている。GATA3のプロモーターは2箇所想定されているので2種をL、Sと命名した。また、活性部位をTFSERCH検索し、活性部位と想定された範囲を欠くクローンを2000baseクローンにあわせて作成した。各プロモーターにより想定される活性部位が異なり、例えばKLK11は GATA1、GATA2、GATA3のいずれもが検索されるが複数箇所あり残念ながら欠落させられなかった。SP1などの主要転写因子は複数箇所存在することが多く、欠落させられないのが残念であった。HSF-2のような1箇所しかない場合やMZF-1の様に近接する2箇所の時はまとめて欠落させた。検索結果の分析に相当の時間を要した。欠落クローンは多岐にわたり、個数も異なる。これらのクローンを皮膚培養細胞NKT1と対照用に骨芽細胞MCH3T3-E1に導入し、発現量の変化の測定を開始した。多くのクローンの作成に時間がかかり、ルシフェレース活性を測定できだしたところで、有効な長さや活性部位の特定には至っていない。また、GATA3を発現させる真核細胞発現ベクターpcDNA3に組み込み、上記2種の細胞に導入した。観察を1週以上行ったが、形態的変化は認められなかった。対象としている酵素群値にも値の変化は認められなかった。研究課題/領域番号:19K10597, 研究期間(年度):2019-04-01 - 2021-03-31出典：研究課題「転写因子・作用蛋白の同定による表皮の分化・脱核機構の解明」課題番号19K10597（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/grant/KAKENHI-PROJECT-19K10597/）を加工して作

Exacerbation of daily cough and allergic symptoms in adult patients with chronic cough by Asian dust: A hospital-based study in Kanazawa

The health effects associated with Asian dust have attracted attention due to the rapid increase in the number of Asian dust events in East Asia in recent years. The aim of this study was to investigate the associations between Asian dust and daily cough, as well as allergic symptoms, in adult patients who suffer from chronic cough. We enrolled 86 adult patients from Kanazawa University Hospital, Japan, who were diagnosed with asthma, cough variant asthma, atopic cough or a combination of these conditions. From January to June 2011, subjects recorded their symptoms in a diary every day. Asian dust and non-Asian dust periods were defined according to the dust extinction coefficient, measured using the light detection and ranging (LIDAR). The daily levels of total suspended particulates, polycyclic aromatic hydrocarbons (PAHs) and coexisting factors related to allergies, such as the Japanese cedar pollen count, were measured. McNemar\u27s test showed that there were significantly more cough-positive patients during Asian dust periods than during the non-Asian dust period (p = 0.022). In addition, during Asian dust periods when the daily levels of Japanese cedar pollen, Japanese cypress pollen and PAHs were elevated, there were significantly more patients who experienced itchy eyes than during the non-Asian dust period (p < 0.05). On the other hand, there were no significant differences in the allergic symptoms, including sneezing or a runny nose and nasal congestion. This is the first report to show that Asian dust triggers cough and allergic symptoms in adult patients with chronic cough. © 2014 Elsevier Ltd. All rights reserved

Effects of Asian dust on daily cough occurrence in patients with chronic cough: A panel study

Asian dust, known as kosa in Japanese, is a major public health concern. In this panel study, we evaluated the effects of exposure to kosa on daily cough occurrence. The study subjects were 86 patients being treated for asthma, cough variant asthma, or atopic cough in Kanazawa University Hospital from January 2011 to June 2011. Daily mean concentrations of kosa and spherical particles were obtained from light detection and ranging (LIDAR) measurements, and were categorized from Grade 1 (0μg/m3) to 5 (over 100μg/m3). The association between kosa and cough was analyzed by logistic regression with a generalized estimating equation. Kosa effects on cough were seen for all Grades with potential time lag effect. Particularly at Lag 0 (the day of exposure), a dose-response relationship was observed: the odds ratios for Grades 2, 3, 4, and 5 above the referent (Grade 1) were 1.111 (95% confidence interval (CI): 0.995-1.239), 1.171 (95% CI: 1.006-1.363), 1.357 (95% CI: 1.029-1.788), and 1.414 (95% CI: 0.983-2.036), respectively. Among the patients without asthma, the association was higher: the odds ratios for Grades 2, 3, 4 and 5 were 1.223 (95% CI: 0.999-1.497), 1.309 (95% CI: 0.987-1.737), 1.738 (95% CI: 1.029-2.935) and 2.403 (95% CI: 1.158-4.985), respectively. These associations remained after adjusting for the concentration of spherical particles or particulate matter with an aerodynamic diameter of less than 2.5μm (PM2.5). Our findings demonstrate that kosa is an environmental factor which induces cough in a dose-response relationship. © 2014 Elsevier Ltd

Parathyroid hormone 1 (1-34) acts on the scales and involves calcium metabolism in goldfish

金沢大学環日本海域環境研究センターThe effect of fugu parathyroid hormone 1 (fugu PTH1) on osteoblasts and osteoclasts in teleosts was examined with an assay system using teleost scale and the following markers: alkaline phosphatase (ALP) for osteoblasts and tartrate-resistant acid phosphatase (TRAP) for osteoclasts. Synthetic fugu PTH1 (1-34) (100 pg/ml-10 ng/ml) significantly increased ALP activity at 6 h of incubation. High-dose (10 ng/ml) fugu PTH1 significantly increased ALP activity even after 18 h of incubation. In the case of TRAP activity, fugu PTH1 did not change at 6 h of incubation, but fugu PTH1 (100 pg/ml-10 ng/ml) significantly increased TRAP activity at 18 h. Similar results were obtained for human PTH (1-34), but there was an even greater response with fugu PTH1 than with human PTH. In vitro, we demonstrated that both the receptor activator of the NF-κB ligand in osteoblasts and the receptor activator NF-κB mRNA expression in osteoclasts increased significantly by fugu PTH1 treatment. In an in vivo experiment, fugu PTH1 induced hypercalcemia resulted from the increase of both osteoblastic and osteoclastic activities in the scale as well as the decrease of scale calcium contents after fugu PTH1 injection. In addition, an in vitro experiment with intramuscular autotransplanted scale indicated that the ratio of multinucleated osteoclasts/mononucleated osteoclasts in PTH-treated scales was significantly higher than that in the control scales. Thus, we concluded that PTH acts on osteoblasts and osteoclasts in the scales and regulates calcium metabolism in goldfish. © 2011 Elsevier Inc. All rights reserved