Glycation levels in serum apo B proteins were measured by colorimetry of fructosamine using nitroblue tetrazolium in immunoprecipitates generated using anti-apo B serum. The serum levels of apo B glycation determined by the proposed method correlated well with the fructosamine concentrations in low-density lipoprotein (LDL) fractions separated by ultracentrifugation. Close correlations were also observed with serum concentrations of fasting blood glucose, total cholesterol and apo B protein, but those with serum triglyceride, fructosamine and HbA1 c were weak. The normal reference value was 18.1±3.65μmol/l and inter-and intra-assay coefficients of variation were 4.6-6.1% and 5.9-6.6%, respectively. Diabetes mellitus patients with vascular complications had higher serum glycated apo B levels than those without complications, matched for risk factors such as blood pressure, obesity, smoking history, age, sex, duration of diabetes, serum total cholesterol, triglyceride, high-density lipoprotein (HDL) cholesterol and lipoprotein (a). High levels of glycated apo B proteins in the blood might be an independent risk factor in addition to the established risk factors for diabetic vascular complications.

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The sugar chains of serum immunoglobulin (Ig) G, A, and M in rheumatoid arthritis (RA) patients (n=48) with elevated and normal C-reactive protein (CRP) were compared with those in age-matched healthy control subjects (n=17). The levels of agalactosylated sugar chains (GO) of serum IgG were increased significantly (p<0.01) in RA patients with elevated serum CRP, while those in RA patients with normal serum CRP remained similar to those in healthy subjects. The digalactosylated sugar chain levels were also decreased significantly (p<0.01) in serum IgA and IgM of active RA patients. Increases in levels of the bisected sugar chains were observed in serum IgG, IgA, and IgM from active RA patients. Furthermore, the GO contents of serum IgG in RA patients were correlated with their serum CRP levels (r=0.5165). These results indicated that disturbed glycosylation of Ig molecules occurs commonly in patients with active RA.

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UV検出器を用いた高速液体クロマトグラフィー (HPLC) による, 人体液中メバロン酸 (MVA) の測定法を開発した。本法は, メバロン酸を抽出乾固する際に少量の強酸を加え, メバロン酸のラクトン体であるメバロノラクトン (MVL) をデヒドロメバロノラクトン (D-MVL) へと変化させ, そのUV吸収を測定することを原理とする。添加回収率は, 血漿で95.6～104.0%, 尿で99回4～103.8%を示した。検出限界は, S/N=4で 0.8ng/mlであった。再現性, 直線性も良好であり臨床検査として有用であると考えられた。

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メバロン酸類縁体を合成し, ハプテンとしてキャリア蛋白に結合し, これを抗原としてマウスに感作しメバロン酸に対して特異的なモノクローナル抗体を得た。この抗体と, ペルオキシダーゼでの標識抗原を用いてメバロン酸のエンザイムイムノアッセイ (EIA) の系を確立した。測定範囲は1.5～170pmol/test (IC₅₀=15pmol/test) であった。また, 生体中に含まれるメバロン酸類似物との交差反応は0.001%以下となり極めて高い特異性が得られた。正常男子 (n=9) にヒドロキシメチルグルタリルーCoA (HMG-CoA) リダクターゼ阻害剤であるプラバスタチンを投与し, その尿中メバロン酸をこの系を用いて定量すると, 全例でメバロン酸値は低下した。このEIAで得られた人尿中メバロン酸の測定値は, より基本的なメバロン酸キナーゼを利用したラジオエンザイム法での測定値との相関において良好 (r=0.969) な結果が得られた。

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乳び胸の早期診断に胸水および血清の脂質 (トリグリセリド, エステル型コレステロール, 遊離型コレステロール, リン脂質) 濃度を測定し, それぞれの比を利用することが有用であった。すなわち, トリグリセリドの濃度比だけが他の脂質の濃度比より大きい場合を乳び胸水と判定することができる。この方法によって, 胸水中トリグリセリドが280 mg/lで, リボ蛋白電気泳動でカイロミクロンが検出されなかった症例において, 胸水/ 血清のトリグリセリド比, エステル型コレステロール比, 遊離型コレステロール比, リン脂質比は, それぞれ0.64, 0回46, 0.47, 0.46と乳び胸を疑わせる結果が得られた。2日後に同症例の胸水のリボ蛋白電気泳動においてカイロミクロンが証明され, 脂質濃度比もそれぞれ1.02, 0.46, 0.45, 0.48と胸水中トリグリセリドの割合だけが増大し, しかも血清トリグリセリド値を上回っており, 明らかに乳び胸と診断された。また胸水中トリグリセリド値が390mg/1で乳び胸でない症例や, それとほぼ同等の380mg/1で乳び胸であった症例もあり, トリグリセリドの絶対値による乳び胸の診断が必ずしも可能でないことから, 本法が乳び胸の早期診断に有効であることが示唆された。

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The recommended method for measuring the activity of serum γ-GT was established by Japan Society of Clinical Chemistry (JSCC). This method was based on the IFCC method, and the final concentration of GluCANA as a donor substrate and that of glycylglycine as an acceptor substrate were set at 6.0 mmol/ 1 and 150 mmol/ 1 respectively. The initial velocity under these conditions was about 77% of V_max. However, it is inappropriate to increase further the concentrations of the substrates, because of unacceptable increasing absorbance of reagent blank.
This method has the following two characteristics. One is that the amount of the sample was set small so as to avoid the turbidity to becaused by the reaction between glycylglycine and the serum, and the other is that the procedure is two-step system in which the reaction is made to start by GluCANA as the donor substrate.
It was confirmed that there was no difference in the measurement value obtained by this recommended method and the IFCC method. Therefore, this method will be used for traceability to the IFCC method. The consensus method, which is derived from this recommended method by setting the reaction temperature at 37°, will be used for traceability of the measured value by the field method. Furthermore, the consensus method is also used for valuating the enzyme reference materials and for checking of commutability.

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