臨床化学
Online ISSN : 2187-4077
Print ISSN : 0370-5633
ISSN-L : 0370-5633
12 巻, 1 号
選択された号の論文の12件中1~12を表示しています
  • ,
    1983 年 12 巻 1 号 p. 1-9
    発行日: 1983/03/25
    公開日: 2012/11/27
    ジャーナル フリー
    Proteins can undergo a variety of post-translational modifications.One of the most widespread is non-enzymatic glycosylation. This phenomenon has gained considerable attention because of its association with diabetes mellitus and the possibility that it may contribute to the long-term complications of the disease. This concise review will focus on non-enzymatic glycosylation of hemoglobin and other proteins and the relationship between this process and diabetes.
  • 第二報, Heparin-Releasabie LPLについて
    楠見 博明
    1983 年 12 巻 1 号 p. 10-23
    発行日: 1983/03/25
    公開日: 2012/11/27
    ジャーナル フリー
    As described in a preceding paper, two lipoprotein lipase bands (b-LPL and f-LPL) were separated from extracts of adipose and other tissues from mice by electrophoresis. In the present paper, some studies with respects to these two types of lipoprotein lipase were made.
    It is well known that LPL activity in serum increases after heparin administration in vivo. This is considered to be originated from cell surface of fat or mugcle through a competitive binding of heparin to the cell surface.
    It was demonstrated the increase in BAT (brown adipose tissue) LPL activity by the coldexposure was canceled by heparin administration in vivo, and LPL activity in PHP (Postheparin plasma) was increased more in coid-exposed mice than in control ones.On the other hand, it was Shown in zymography that the f-LPL and b-LPL bands from BAT-extracts under cold exposed-mice were strongly stained, furthermore, their bands were decreased by injection of heparin, and the decrease of b-LPL band was especially apparent.The b-LPL band in PHP was increased more in cold exposed-mice than in control ones.Here, the b-LPL band in PHP from Cold exposed-mice was stained more strongly than that from control ones.
    Since the increase of LPL could not be observed in other tissues except BAT of animals stressed by the cold, most parts of the increased activity in plasma was considered to be originated from BAT.From results of studies on relationship between b-LPL band and f-LPL band, it seemed most reasonable to conclude as follows.Namely, the b-LPL band in PHP was one type of LPL which was bound to be administrated heparin, and was originated from LPL located on the cell surface.On the other hand, the b-LPL band in tissue-extracts was an extracellular LPL bound to heparin-like glycosaminoglycan on the capillary endothelial surface, so called“heparin-releasable LPL”.lt was believed to be a functional LPL.
    Furthermore, the f-LPL which was an another type of LPL seemed to be a glycosaminoglycan-free, an intracellular LPL.
  • 1983 年 12 巻 1 号 p. 23-
    発行日: 1983年
    公開日: 2012/12/14
    ジャーナル フリー
  • 第三報, LPL活性に及ぼすヘパリン, プロタミンおよびイオン濃度による影響
    楠見 博明
    1983 年 12 巻 1 号 p. 24-29
    発行日: 1983/03/25
    公開日: 2012/11/27
    ジャーナル フリー
    A new method of lipase-electrophoresis was developed and two types of LPL (lipoprotein lipase) were separated from tissue-extracts by zymography as previously reported 7-8). The one was namedb-LPL. lt was binding LPL with heparh-like glycosaminoglycan whlch might be existing on the cell surface. Another type was named f-LPL. It was free-LPL, which was not bound with glycosaminoglycan. ln the present paper, a method of fractionated-determination of these two types of LPL have been established utilizing their different properties.
    LPL was purified from postheparin plasma by heparin affinity column chromatographic technique. The fractionated b-LPL and f-LPL were prepared respectively by addition of heparin (25u/ml) or protamine sulfate (100μg/ml).Each LPL were shown different enzymatic activity one another at different ionic strength in assay mixture. Namely, the b-LPL had not enzymatic activity at low ionic strength, but was shown a maximal activity at higher ionic strength.On the contrary, f-LPL type was shown a maximal activity at lower ionic strenght.
    On the other hand, several lipases other than two types of LPL were contained in clude extract from tissue, however, two types of LPL are distinguishedl from these lipases by: dependence for activity on a serum cofactor and inhibitiion by 1 mol/l NaCl concentrations. Furthermore, the serum activated-lipase which was determined at low ionic strength in assay mixture was not b-LPL, but f-LPL type. Since the b-LPL type was converted to f-LPL type by addition of protamine sulfate to enzyme mixture, the serum activated-lipase (total LPL activity) was equal to sum of f-LPL and b-LPL activity. The b-LPL activity, consequently, enable to be estimated by subtracting the f-LPL activity determined at low ionic strength from total LPL activity.From these results, it was emphasized to benecessary to Pay much attention to ionic strength in assay mixture for determination of LPL activity.
  • 太子 馨, 戸沢 辰雄
    1983 年 12 巻 1 号 p. 30-38
    発行日: 1983/03/25
    公開日: 2012/11/27
    ジャーナル フリー
    Enzyme-immunoelectrophoresis and enzyme-immmofixation on agar-agarose gel or cellogel membrane are usually used for the identification of alkaline phosphatase (ALP)-linked immunoglobulin (lg).However, there are some cases which could not be detected by these methods.
    We applied the immunoprecipitin reaction in free liquid media for the sensitive method of the identification of ALP-linked lg. By this method, it is easy to determine existence of ALP-linked lg from ALP activity in immunoprecipitate formed by each monospecific antiserum. ln the presernt study, ALP-linked lgG (λ) was identlfied in all of 6 cas es by this method. The result corresponded with that from immunoelectrophoresis.Besides, we observed the change of ALP-linked lgG in clinical course and could confirm the sensitive method, by which ALP-lhked lgG in every case could be detected even that could not be detected by enzyme-immunoe-ectrophoresis.
    High sensitivity of this method for the detection of ALP-linked lgG comes from that many precipitates were obtained and from that procedure time was short enough to avoid loss of ALP activity.
    As ALP bound to lgG in almost cases, we also tested the method using Protein A-Sepha rose CL-4B.However its detective sensitivity was most, ALP-linked lgG3, lgA and lgM could not be identified.Therefore, the immunoprecipitin reaction was demonstrated as the simple, quick and reliable method to identify both class and type of ALP-lhked lg.
  • 谷島 清郎, 本間 啓子, 長原 三輝雄
    1983 年 12 巻 1 号 p. 39-44
    発行日: 1983/03/25
    公開日: 2012/11/27
    ジャーナル フリー
    The addittion of ethylene glycol or glycerol to pooled human serum protects lactate dehydrogenase (LDH) activity during storage at-10°C or 4°C.At-20°Cethylene glycol caused a significant decreasein LDH activity during storage, the decrease depending on the proportion of added ethylene glycol.Reduced activity was accompanied by loss of the LDH3, LDH4 and LDH5 isoenzymes.ln contrast, glycerol addition to serum completely stabilizes LDH activity even at-20°C, and concomitant measurement of alkaline phosphatase, glucose and cholesterol showed no alteration in activity or concentrations.
    Glycerol treated pooled serum would thus seem valuable for quality control for clinical use.
  • 奥田 彰洋, 橋本 聡一, 實川 佐太郎, 川島 康生
    1983 年 12 巻 1 号 p. 45-53
    発行日: 1983/03/25
    公開日: 2012/11/27
    ジャーナル フリー
    A gas chromatographic method for the quantitive determination of oxygen and carbon dioxide contents in the blood was investigated.A Shimazu gas chromatograph (GC -4CIT) and a Shimazu blood gas sampler (BGS-1A) were used for the analysis.
    Oxygen and carbon dioxide were freed from blood with Van Slyke-Neil's reagent in BGS-1A and were directly transfered into gas chromatograph and digital integrator (Chromatopack-E1A) was used for calculating gas chromatographic peak area.
    The reproducibility of the method was 0.8% (coefficient of variance) wlth the analysis of oxygen content and was 0.5% (C.V.) with the analysis of carbon dioxide content respe ctively.Agreement between gas chromatographic and Lex-02-Con values for oxygen content was excellent (r=0.980) and also excellent agreement was recognized for the analysis of carbon dloxlde content between gas chromatographic values and those obtalned from Van Slyke-Sendroy's nomogram (r=0.944).
    ln this study, it wsa proved that this gas chrdmatographic method for the analysiis of oxygen and carbon dioxide contents in the blood was accurate enough to obtain satisfactory measurements and imposed relatively little skill and training in contrast to the conventional Van Slyke-Neil's manometric method.
  • 三浦 順吉, 宮城 宏行, 高田 芳矩, 山形 陽
    1983 年 12 巻 1 号 p. 54-62
    発行日: 1983/03/25
    公開日: 2012/11/27
    ジャーナル フリー
    A clinical liquid chromatograph, which consists of a completely automated liquid chromatograph combined with a microcomputer for diagnosis, and it's application to rerlal function analyses are described.The analytical rate for urinary ultraviolet-absorbing constituents using anion-exchange chromatography was twelve samples per day.The reproducibilities of retention time, peak area and absorbance ratio (A23nm/A250nm) were less than 1%, 5%, and 6%, respectively for continuous five days running.
    Approximately 90 morning fasting urhes were obtained from adult“healthy”subjects, inpatients and outpatients.The urine samples were stored at-20°C until used, and defrosted at room temperature in a water bath at 37°C. They were centrifuged and then the supernatant was filtered through a 0.1μm pore size membrane filter for the chromatography.
    Each chromatographic peak was numbered in accordance with peak identification parameters such as retention time and absorbance ratio.These peaks were rearranged in the order of peak number instead of retention time.
    The reconstructed chromatograms were used to discuss kidney functions in three ways. Those are (1) comparison of chromatograqhic profiles between normal and abnormal subjects,(2) correlation between creatinine clearance and a chromatographic peak,(3) correlation analysis of peak area between two specific peaks.
    These methods as mentioned above seem to be usefull for the estimation of pathological phases and metabolic change induced by drugs and diseases.
  • 荒尾 雅代
    1983 年 12 巻 1 号 p. 63-78
    発行日: 1983/03/25
    公開日: 2012/11/27
    ジャーナル フリー
    Serum high density lipoprotein (HDL) fractions of various hyperlipidemic patients were separated by polyanion methodand ultracentrifugation, then their lipid compositions and apolipoprotein A (apo A) were analyzed.
    HDL cholesterol of hypertriglyceridemic patients were significantly lower than those of normolipidemic subjects.On the other hand, apo A in HDL of Type IIb patients were not different from those of normotriglyceridemias.Also in other types of hypertriglyceridemias, apo A in HD were not significantly lower.
    The ratios of apo A/total cholesterol and apo A/esterified cholesterol in HDL from hypertriglyceridemias were higher than those from normotriglyceridemias and the ratios of apo A/triglyceride in HDL from hypertriglyceridemias were lower.Decrease of cholesterol and increase of triglyceride were observed in HDL fractions from hypertriglyceridemias and hypo α-ipoproteinemias.
    Correlation coefficient between HDL concentration and HDI apo A was better than that between HDL and HDL cholesterol.
    VLDL fraction was increased significantlay nd HDL fraction was decreased in Triton induced hypertriglyceridemic rats and similar compositional change of HDL (the relative increase of triglyceride and decrease of esterified cholesterol) Was observed.
    From the results of analysis of apoprotein in HDL and VLDL from Triton injected rats by isoelectric focuslng, it was shown that apo C in VLDL was significantly increased and apo C in HDL was decreased.
  • 宇佐美 英治, 野村 喜重郎, 内田 寛, 瀬山 義幸, 山下 三郎
    1983 年 12 巻 1 号 p. 79-85
    発行日: 1983/03/25
    公開日: 2012/11/27
    ジャーナル フリー
    An oral elastase preparation (Elaszyme, Eisai Co., porcine pancreatic) was administered to 42 patients with hyperlipemia having more than 220mg/dl of total cholesterol and more than 120 mg/dl of triglycerides.
    Although oral enzyme preparation may be poorly absorbed into the blood stream, serum elastase activity, on suc (Ala) 3pNA, 14C-elastin and autoclave treated natural elastin, showed an increasing tendency by the administration of the preparation.Significantly higher serum levels of total cholesterol, triglycerides, phospholipids, ζ-lipoproteins and lipid peroxides were lowered to the extent of the normal levels, and such normalizing tendency was more remarkable when these levels were considerbly higher before the administration of the preparation.
  • 岩井 弘行, 奥出 政義, 秋浜 澄行
    1983 年 12 巻 1 号 p. 86-88
    発行日: 1983/03/25
    公開日: 2012/11/27
    ジャーナル フリー
    A sensitive fluorometric determination of peroxidase activity using o-phenylenediamine as substrate is described.
    Upon oxidation with peroxidase and hydrogen peroxide, o-phenylenediamine is converted to the2, 3-diaminophenazine which has an excitation maximum at 410nm and an emission maximum at 550 nm in neutral solution.Under the optimum conditbns, peroxidase was possible to determine in the range of 0.62 to 31.23μU/ml.
  • 笠原 靖, 藤井 信之, 水越 幹雄, 永津 俊治
    1983 年 12 巻 1 号 p. 89-93
    発行日: 1983/03/25
    公開日: 2012/11/27
    ジャーナル フリー
    Multiple forms of glycylprolyl dipeptidyl-amhopeptidase (GP-DAP) were analyzed in human serum, liver, kidney, spleen, and lymphocytes by our methods based on electrophoresis with fluorometry using 7-glycyl-proline-4-methylcoumarinamide tosylate as substrate.Four peaks of the activity of GP-DAP were separated by the electrophoretic analysis.The main peak (peaklll) in normal human serum was found to coincide with that in the human spleen and lymphocytes.Peak ll and peak IV enzymes, which were predominantly observed in human sera from patients with hepatitis, coincided with the main peaks of crude and pure enzymes in the liver and kidney, respectively.
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