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Presently we, Keio Endocrine and Metabolite Survey (KEMS) study group conducted a questionnaire sur- vey with respect to panic values in the laboratories belonging to Keio University-associated hospitals. As to the initial setting, most of the laboratories answered to play a leading role in preparing the necessary matters to implementation of panic values and revise them corresponding to physician's request on each occasion. In almost all laboratories, they did not verify whether the notification procedure does work to exert appropriate clinical action. The numbers of critical values answered by the 18 laboratories distributed widely in the test items (8-39) and their critical limits (10-68). As to the critical limits, the lower limits of serum K and blood glucose converged among the laboratories, however, the limits of other test items diverged. The results of the present survey regarding to critical values, although being in small scale, may submit the im- portant issue to be solved in near future. [Short Communication].

We examined the correlations between serum dolichol levels and laboratory test parameters in patients affected by disease, as well as the distribution of dolichol in sera from patients with hyperbetalipoproteinemia and hyperalphalipoproteinemia. Serum dolichol was evaluated by a reverse-phase HPLC method. After centrifugation, the serum dolichol found in healthy controls was mainly associated with medium-sized particles of the high-density lipoprotein (HDL) fraction. For patients with hyperbetalipoproteinemia, serum dolichol was also associated with the medium HDL fractions. However, for hyperalphalipoproteinemia patients the levels of large HDL and serum dolichol were increased, and serum dolichol was mainly associated with the large HDL fraction. On laboratory tests of components, the dolichol level was not correlated with the values for markers of the liver and biliary system, with the values of renal function markers, with creatine kinase activity, amylase activity or uric acid concentration, but was correlated with total cholesterol, HDL-cholesterol and apoA-I concentrations, and with lactate dehydrogenase (LDH) activity. These results suggest that serum dolichol exclusively localized in HDL, and in subpopulation, that in normocholesterolemia or hyperbeta-cholesterolemia is associated with HDL(3), which is small sized and high density HDL, however, that in hyperalphacholesterolemia is associated with HDL(2), which is large sized and lower density HDL.

ADH/GPT比が高値を示した成人発症Still病(AOSD)の2例を報告する.症例1は30歳女性.39℃台の弛張熱,発疹および関節痛を主訴に当院を受診.GOT 133IU/l, GPT 417IU/l, LDH 514IU/lと肝機能障害が認められ入院した.ADH/GPT比が高値を示し,さらに急性期の肝生検像で,中心静脈周囲に限局した肝細胞壊死が確認された.症例2は重症肝障害および播種性血管内血液凝固症を合併した44歳女性.ADH/GPT比が高値を示した.いずれの症例も副腎皮質ステロイド薬が著効し軽快した.これらAOSD 2例における肝障害の主座は中心静脈領域であることが示され,さらに血清ADH活性測定は肝障害の鑑別にも有用と考えられた.

膜の荷電がゼロで電気浸透度のまったくない新しいセ・ア膜『SEPARAX-SP (nSP)』での健常者の血清蛋白分画の基準値を設定した. 支持体の電気浸透度, 塗布位置や膜構造の均質性などの物性が変わったことにより, その基準値は, 塗布点がγ分画の中央にある Sartorius (SR) 膜での値と類似の結果であったが, α2とβ分画には相違があった. 一方, SEPARAX (SP)膜とはすべての分画で異なった値を示した. この原因として, βリポ蛋白質がSP膜よりブロードな分布をとることと, nSP膜の血清塗布位置がγ分画をはずれた陰極側になったため, albなどの先行蛋白の吸着による影響の可能性が波形解析データから示唆された. 新しいnSP膜による血清蛋白分画値は, 自動機法や用手法などの測定方法による差はほとんどなく, SP膜で指摘されていた問題点も改善され, 日常検査への適用は有用と思われた.

自動電気泳動装置で得られる蛋白分画像の情報を中心とした, エキスパートシステムの構築のための基礎検討を行った. 装置の改良と進歩により再現性のよい分画像が得られるようになり, 従来, 経験により判読されている電気泳動波形データの解析をマイクロコンピュータを用いて定量的, 視覚的に行えるシステムを目標とした. 562検体を用いて, 健常人の基準泳動波形を求めた. この健常人の波形の上に被検体波形を重ね書きする方法により, 量的な変動を一目で捉えることができた. 定量化法は, 変動を各項目共通の単位として取り扱えるSDインデックス (測定値-平均値)/標準偏差法を考案し, データの標準化を行った. 電気泳動関連パラメータとして17項目 (5分画%, A/G比, TP値, Alb分画半値幅, 各分画のピークおよび谷の位置のモード的易動度) を選択し, それらの変動の分布をSDインデックスで検討したところ, 異なる健常人集団の95%以上が±3SDに含まれ, ±3SDが異常値と正常値の目安となることが判明した. 本手法を定型的な病的血清蛋白症例に応用したところ, 泳動波形とSDインデックスのグラフィック表示により各病型の特徴を容易に捉えることができた.

A 28 year old woman with Hashimoto's disease was treated with desiccated thyroid and triiodothyronine (T3). She improved steadily during the first 2 to 3 months and thyroidal function tests turned to normal. Then, in spite of continuing treatment, her serum T4 level decreased gradually and she became fatigued. A serum T3 radioimmunoassay manifested an interference pattern suggested anti-T3 antibody in her serum. Ethanol-extracted serum T3 and T4 levels were low in spite of ingestion of desiccated thyroid or synthetic T3 and T4, suggesting intestinal malabsorption of T3 and T4. Antibodies against T3 and T4 were identified in her serum; affinity constants were 1.16 X 10(10) and 8.73 X 10(8) l/mol respectively. After treatment with synthetic T3 and/or T4 for 20 months, the titer of anti-T3 and anti-T4 antibodies decreased, and impaired intestinal absorption of thyroid hormone improved. Then, after desiccated thyroid treatment was reinstituted, the anti-T3 antibody titer again increased and intestinal absorption of thyroid hormone decreased. These results suggest the oral immunization against thyroid hormones. There was associated impairment in intestinal absorption of thyroid hormone presumably secondary to the anti-T3 and anti-T4 antibodies.

Many cases of macromolecular amylases were reported. Various modes of complex formation with normal amylase were considered to be categorized as follows: 1. a polymer complex of normal amylases, 2. a complex of normal amylase and plasma proteins other than immunoglobulins (e. g., some glycoproteins), 3. a complex of normal amylase and some other large non-protein molecules (e. g., glycogen or other mucopolysaccharides) and 4. a complex of amylase and immunoglobulins.In this study, four cases of amylase-linked immunoglobulins were reported and the mode of binding form between amylase and immunoglobulins were discussed.Three of the cases were tumor-bearing patients of lung, gastric and esophageal cancer, and therapeutically, pancreatic enzyme preparations were not administered in the past three years. The last one was a patient with liver cirrhosis and chronic pancreatitis who have not been administered with pancreatic drugs.The class of heavy chain of amylase linked immunoglobulin was proved to be γ in three and α in one of the cases by immunoelectrophoresis followed by amylase activity staining. The type of light chain was determined to be exclusively λ irrespective of heavy chain class.In three of the cases, the immunoglobulin complexes were partially dissociated into normal amylase and immunoglobulin G at pH8.6, and completely dissociated at pH9.0. These phenomena of dissociation might give a clue to explain the heterogeneity of macromolecular amylases which have been discussed previously.Papain digestion of amylase linked immunoglobulin G was carried out to elucidate the specific antigen-antibody bindings of these macromolecular complexes. The molecular size of the digest was determined to be smaller by thin layer gel-filtration. The precipitin line formed against anti-Fab antiserum was proved to have amylase activity, but that against anti-Fc was not. This fact suggests that the binding site of amylase linked immunoglobulin G was located in Fab portions and that the complexes are specific antigen-antibody complexs.Thus, it was elucidated that the complex formation of amylases and immunoglobulins in blood plasma is one of the circulating autoantibodies and must be clearly distinguished from the other unknown macromolecular amylase complexes.

The combination of both the electrophoresis and the immunofixation and of the thin layer gel filtration and the immunofixation followed by enzyme activity staining on antigen-antibody complexes were applied to detect and identify the enzyme-linked immunoglobulins which were observed as macromolecular enzyme complexes in human sera. For any given antigen-antibody system, the optimum concentration of antigen in the serum to be analysed must be given experimentally. Moreover, removing of unreacted proteins must be carefully performed with the solution which will protect the activity of the enzyme to be detected.In the cases of LDH- and amylase-linked immunoglobulins the detection of minimum enzyme activities on Cellogel membrane were limited by staining sensitivity. Thus, two to ten times concentration of commercially available antibodies was required to give the optimum concentration of antigen-antibody complexes. In the cases of alkaline-phosphatase-linked immunoglobulins, the detection of the enzyme activity was easily amplified by the elongation of the incubation period. Thus, the optimum concentration of antigen was given without concentration of commercial antibodies.In the studies of macromolecular enzyme complexes in human sera, the immunoprecipitation technique and enzymo-immunoelectrophoresis were conventionally carried out to identify the enzyme-linked immunoglobulins. Some cases, however, still ramain ambiguous because of unskilled and unstable techniques used for analysing these macromolecular enzyme complexes. Thus, these immunofixation techniques followed by enzyme activity staining (enzyme immunofixation), described in this paper can facilitate the detection and identification of such macromolecular complexes as enzyme-linked immunoglobulins.

The production of early and term placental alkaline phosphatase (ALP), human chorionic gonadotropin beta-subunit (beta-hCG) and pregnancy-specific beta 1-glycoprotein (SP1) was confirmed in the newly established uterine cervical cancer call line SKG-IIIa. Treatment of these cells in culture with sodium butyrate caused an increase of all of these oncodevelopmental proteins. On the other hand, prednisolone treatment enhanced only term placental ALP, while it reduced early placental ALP, beta-hCG and SP1. These data suggested that such oncotrophoblastic proteins as early placental ALP, beta-hCG and SP1 were concordantly modulated by butyrate and prednisolone. These findings may support the possibility of the reexpression of sets of development phase-specific genes in cancer cells.

The dexamethasone suppression test (DST) was administered to 59 patients with major affective disorders and 19 with other psychiatric disorders. The combinations of two dexamethasone doses (1mg vs 0.5mg) and two cortisol criterion levels (5μg/dl vs 4μg/dl) were adopted. The highest sensitivity (43%) was found with 0.5mg DST at the 4μg/dl criterion level for Major Depression. Our study failed to distinguish Melancholia in DSM III.

The dexamethasone suppression test (DST) was performed in 38 patients satisfying the Research Diagnostic Criteria (RDC) for cortisol, human growth hormone (hGH), prolactin, triiodothyronine (T3), thyroxine (T4) and thyroid-stimulating hormone (TSH) were determined at 4:00 p.m. before the administration of dexamethasone, whereas cortisol, hGH and TSH were determined at 4:00 p.m. the day after. Among hormonal variables, only cortisol index (the ratio of the pre-DST cortisol to the post-DST cortisol) was barely useful for discriminating depression. The cortisol index was also correlated with RDC incapacitating subcategory and with symptoms of affect such as depressed mood and psychic anxiety.

We describe an improved method for determination of alcohol dehydrogenase (EC 1.1.1.1) activity in 60 microL of human serum, based on conversion of ethanol to acetaldehyde with simultaneous reduction of NAD+ in glycine NaOH buffer (pH 9.0) at 37 degrees C in a centrifugal analyzer. The final concentration of NAD+ was 10 mmol/L and ethanol was 20 mmol/L. The dilution curve was linear with enzyme activity up to 200 U/L, and results by this method correlated well with those by a manual method (N Engl J Med 279: 241-248, 1968). Within-run precision (CV) was 0.9 to 8.2% over the range of 4.5 to 88.1 U/L, and day-to-day precision was 5.4 to 5.6%. In sera from 198 healthy individuals, mean alcohol dehydrogenase activity was 1.6 (SD 1.2, range 0-5) U/L. To evaluate the clinical utility of determining alcohol dehydrogenase, we measured the activity of alanine aminotransferase and alcohol dehydrogenase in sera from 470 patients with various diseases in our hospital, and found that results for the two enzymes did not correlate well.

Urinary calculi found in 4 patients on chronic hemodialysis or continuous ambulatory peritoneal dialysis (CAPD) were identified as protein calculi by infrared spectroscopic analysis. Positive Congo red staining and immunological assessment revealed that the calculi were composed of amyloid protein derived from beta 2-microglobulin. A comparison of the patients who excreted calculi with 10 patients on chronic dialysis without urinary calculi showed no significant differences in the urinary and serum levels of beta 2-microglobulin. The mechanism of amyloid calculus formation may involve factors independent of the concentration of beta 2-microglobulin in urine or serum. Urinary calculi found in patients on chronic hemodialysis or CAPD were composed of amyloid protein derived from beta 2-microglobulin.

To determine whether serum alcohol dehydrogenase (ADH) activity reflects hepatic damage of centrilobular region (zone 3), the rats were given either bromobenzene (BB) or allyl alcohol (AA) IP to produce the pericen tral or periportal necrosis respectively. After AA or BB serum alanine aminotransferase (ALT) activity showed no significant difference between the two groups. By contrast, serum ADH and glutamate dehydrogenase (GLDH) activities were elevated preferentially in the BB treated rats. However, AA administration to rats also resulted in a significant increase in GLDH activity, whereas ADH activity was only slightly elevated when compared to controls. Moreover, acute ethanol administration to rats resulted in a significant elevation of the serum ADH activity, whereas serum GLDH and ALT activities remained normal. These data suggest that serum ADH activity appears to be a sensitive and specific marker of hepatic centrilobular damage.

Among 37 patients with DSM-III major depression, the nonsuppression rate of the dexamethasone suppression test was significantly higher in patients with recurrent and melancholic major depression (83%) than in those with single episode and/or nonmelancholic major depression (31%).