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Recently, azole-resistant Aspergillus fumigatus containing a 34-bp or 46-bp tandem repeat in the promoter region of cyp51A combined with amino acid substitution(s) has appeared in the environment worldwide, including several Asian countries. In this study, we isolated the 34-bp tandem repeat-containing azole-resistant A. fumigatus strain OKH50 from a patient in Japan in May 2016. The patient had not been treated with medical azoles before the strain isolation, suggesting that the resistant property was acquired before infection. In addition, the patient had not traveled overseas. Our analysis of short tandem repeats of the strain indicates that the strain is strongly related to the 34-bp tandem repeat-containing isolates from European countries and Asia-Oceania countries but not to susceptible isolates from Japan, suggesting that the strain was introduced from overseas and might spread in Japan.

Azole antifungals are used not only clinically for fungal infections but also used as agricultural fungicides. Recently, azole-resistant Aspergillus fumigatus containing a tandem repeat in the promoter region of cyp51A combined with amino acid substitution(s) appears in the environment in Eurasia, especially in several European countries. Although azole fungicides have been used in Japan, especially in Hokkaido, surveillance and characterization of A. fumigatus in Hokkaido have not been reported. In this study, we collected soil samples from farms that used an azole fungicide in the Tokachi area of eastern Hokkaido, isolated 91 A. fumigatus strains, and determined the minimal inhibitory concentrations of medical azoles required for these strains. Moreover, because causative agent A. fumigatus is ubiquitous in the air and acquired from the environment, we collected 22 clinical isolates of A. fumigatus to measure their susceptibility to medical azoles in a hospital in the Tokachi area. Our data show that almost all A. fumigatus isolates retained susceptibility to medical azoles. Clinical isolates OKH34 and OKH6 showed 8 and 2 μg/mL of voriconazole, respectively, as the minimal inhibitory concentration. Both isolates did not contain tandem repeat in cyp51A promoter region. An isolate contained G448S mutation in cyp51A conferring voriconazole resistance, which is the first report from Japan. Our data shows the existence of azole-resistant and low azole-susceptible clinical isolates and highlight the necessity for continuous surveillance in Japan because resistant A. fumigatus strains can arise through clinical or environmental selection or could be introduced from overseas.

Recently, azole-resistant Aspergillus fumigatus containing a 34-bp or 46-bp tandem repeat in the promoter region of cyp51A combined with amino acid substitution(s) has appeared in the environment worldwide, including several Asian countries. In this study, we isolated the 34-bp tandem repeat-containing azole-resistant A. fumigatus strain OKH50 from a patient in Japan in May 2016. The patient had not been treated with medical azoles before the strain isolation, suggesting that the resistant property was acquired before infection. In addition, the patient had not traveled overseas. Our analysis of short tandem repeats of the strain indicates that the strain is strongly related to the 34-bp tandem repeat-containing isolates from European countries and Asia-Oceania countries but not to susceptible isolates from Japan, suggesting that the strain was introduced from overseas and might spread in Japan.

Chemical contamination in river, ground water and sea was investigated from 1988 to 1992. Water and sediment samples collected in Tokyo area were analyzed by GC/MS, GC and HPLC. Over 40 chemicals were detected in water and/or sediment. Several distribution patterns of chemicals were observed in water and sediment media. In ground water, high levels of volatile chlorinated hydrocarbons were detected. Water and sediment samples from industrial area contained higher concentrations of chemicals than those from urban area. Pesticides and combustion products were observed in wide area. Contamination levels of each chemical in water and sediment samples indicated to be dependent on the physico-chemical property as well as the emission amount.

The ten human cell lines derived from glioma tissues were characterized for the aspects of the phenotypic expression of GFAP and S-100 protein, DNA-ploidy, doublhg time, and response to DBc-AMP and LAK cells. The expression of the TfR was evaluated on these established cell lines with correlation to their phenotypic markers and characteristics. As a result of immunohistochemical studies on frozen materials using a monoclonal anti-Tfit antibody, the high expression of the TfR was related to the doubling time (36-48 hr), but not to the phenotypic expression of glial markers (GFAP and S-100 protein) and other characteristics. And thus, rapidly proliferating astrocytic cell lines including C6 and U251 showed the high level of the TfR in the cells.For the flow cytometry (FCM), the cell harvest with trypsin abolished the extracellular part of TfR molecules from the cell membrane, and thus, single cell preparation with trypsin was not appropriate for the study. On the other hand, EDTA (0.02%) with mechanical shaking provided a preservation of the TfR, molecules for FCM study. The series of FCM analysis demonstrated that newly synthesized (or recycled) TfR molecules appear in the extracellular matrix of the cell membrane by 12 hr and then increase in quantity up to 28 hr after trypsin digestion.The results suggest that the TfR molecules are functioning in rapidly proliferating cells derived from the central nervous system as described previously in other types of tissue.

緑および赤ピーマン中の脂質成分の同定を行なうために, 細切したピーマンは, アセトン中で粉砕した。これをエーテルで抽出し, 全脂質成分を得た。全脂質成分はケン化後, エーテル層と水層とにわけ, 不ケン化物が含まれるエーテル層は, GC分析を行なった。一方, 水層は, 常法に従って脂肪酸エチルエステルを調製し, 脂肪酸組成を調べた。さらに赤ピーマンの全脂質成分は, 薄層クロマトグラフィーによって, キサントフィル類を取り除き, セライト545-酸化マグネシウムで分画した。溶出した各フラクションはケン化後, n-ヘキサンで抽出してGC分析を行なった。各フラクションの主成分は, 再クロマトグラフィーによって単離精製し, MS, IRおよび1H-NMRを測定した。結果は次のとおりである。1) 赤および緑ピーマンの不ケン化物から, α-トコフェロール, カンペステロールおよびβ-シトステロールが, GCによって検出された。α-トコフェロールについては, 緑ピーマンに比べ, 赤ピーマンに多くみられた。2) 総脂肪酸のうち不飽和脂肪酸は, 緑ピーマンが80.8%, 赤ピーマンが66.2%であった。不飽和脂肪酸の主成分は, C18: 2およびC18: 3で, その他にC18: 1がわずかに含まれていた。飽和脂肪酸としては, C12: 0, C14: 0, C16: 0, C17: 0およびC18: 0がみられた。3) 赤ピーマン不ケン化物成分として, イソホロン, 6-ヒドロキシ-4, 6-ジメチル-3-ヘプテン-2-オン, フィトールおよびゲラニルゲラニオールが単離同定された。これらの成分は, 緑ピーマンの不ケン化物中にも検出されたが, ゲラニルゲラニオールの量はわずかであった。