Specificity and polymorphism of diaminopeptidase IV in normal and neoplastic Tμ lymphocytes

Summary In this paper evidence is given for the selective occurrence of the enzyme Diaminopeptidase IV (DAP IV) in human T lymphocytes. This cell population was isolated from human blood and subjected to a cytochemical enzyme reaction. Extending this finding to T-cell leukemias shows that these leukemias can be classified as T cell-derived on the basis of cytochemical tests as well as comparing enzyme polymorphism.Abbreviations DAP IV Diaminopeptidase IV - AcE -naphthyl-acetate-esterase - IEF isoelectric focusing - T-CLL chronic lymphocytic leukemia of T type - B-CLL chronic lymphocytic leukemia of B type     

Lactate dehydrogenase isoenzymes in tumours of the nervous system

Summary The variations in the isoenzymes of lactic acid dehydrogenase have been studied in a series of tumours of the nervous system. From these the ratio of heart muscle component to skeletal muscle component (the H/M ratio) has been calculated and compared with the H/M ratio of a variety of regions of normal brain. Oligodendrogliomata were found to have a very high H/M ratio. There was a decrease in the ratio with increasing degrees of de-differentiation of all tumours studied. However the H/M ratio does not appear directly related to the rate of growth irrespective of the tissue of origin of the tumour. Tumours derived from different tissues appear to have their own range of ratios. Tumour cyst fluids showed similar ratios to their parent tumours.     

Enzyme polymorphism in the classification of human malignant lymphoma

Summary Recent studies on the polymorphism of lysosomal hydrolases have shown that all individual blood cell types in the human being possess their own isoenzyme pattern. In the present study acid phosphatase activity of normal human B-lymphocytes and of four different types of low-grade malignant non-Hodking's lymphomas according to the Kiel classification was estimated. In addition, the isoenzyme pattern of AcP was investigated by isoelectric focusing. Chronic lymphocytic leukemia of B-cell type (N=9) and centroblastic centrocytic follicular lymphoma (N=10) demonstrated significantly lower values than lymphoplasmacytic/lymphoplasmacytoid lymphomas (N=28) and plasmacytomas (N=8). The isoenzyme pattern of normal human B-lymphocytes comprised 12 bands between pH 6.3 and 3.85. This basical pattern was shared by all four lymphoma entities. Only lymphoplasmatic/lymphoplasmacytoid lymphoma and plasmacytoma revealed additional bands, which probably account for the higher net enzyme activity in these cases.Abbreviations AcP acid phosphatase - B-CLL chronic lymphocytic leukemia of B-cell type - CB/CC centroblastic/centrocytic follicular lymphoma - IC lymphoplasmacytic/lymphoplasmacytoid lymphoma - PC plasmacytoma - IEF isoelectric focusing Supported by the Deutsche Forschungsgemeinschaft, programs CL 3 and CN 2     

阻断肾素-血管紧张素系统对糖尿病大鼠肾脏蛋白激酶CβⅡ亚型的影响

目的　研究链脲佐菌素诱导的糖尿病大鼠肾组织蛋白激酶C(PKC) βⅡ亚型的表达和转位 ,以及阻断肾素 血管紧张素系统 (RAS)对其的影响。方法　糖尿病模型大鼠随机分为伊贝沙坦组 (4 0mg·kg-1·d-1)、福辛普利组 (4 0mg·kg-1·d-1)、两药合用组 (各 2 0mg·kg-1·d-1)、糖尿病对照组。另设正常对照组。给药 4周末检测各组血糖、胰岛素水平 ,采用免疫组化法检测肾组织中PKCβⅡ的表达 ,Western印迹法检测肾皮质、髓质PKCβⅡ的表达和胞膜转位情况。结果　糖尿病对照组肾皮质总的和胞膜部分的PKCβⅡ表达量均明显降低 ,分别为正常对照组的 66.0 %和 5 0 .0 % (均P <0 .0 5 ) ,而各治疗组均可部分地纠正这种异常。肾髓质各组间PKCβⅡ的总量及胞膜、胞浆部分的量差异无显著性。结论　早期糖尿病大鼠肾脏中PKCβⅡ的表达存在着量和转位的异常 ,而阻断RAS可部分纠正这种变化 ,提示RAS可通过影响PKC转导途径的异常而参与糖尿病肾病的发生发展机制。     

血管内皮生长因子、端粒酶、腺苷脱氨同工酶联合检测在胸腔积液鉴别诊断中的价值

目的探讨血管内皮生长因子(VEGF)、端粒酶、腺苷脱氨同工酶(ADA)联合检测在良恶性胸腔积液中的诊断价值。方法应用ELISA法检测VEGF的浓度、采用聚合酶联反应-酶联免疫吸附分析法(PCR-ELISA)检测胸腔积液端粒酶活性、用比色分析法检测胸腔积液ADA含量。结果恶性及结核性胸腔积液组VEGF值分别为(327±152)pg/L和(35±15)pg/L,结核组显著低于恶性组(P<0.01)。恶性胸腔积液中端粒酶活性显著高于结核性胸腔积液(P<0.01)。结核性胸液组ADA含量为(45.78±12.78)u/L,高于恶性胸液组(13.56±4.91)u/L,两者间差异有统计学意义。结论检测胸腔积液端粒酶、VEGF和ADA同工酶对癌性胸腔积液的诊断均有一定的价值,联合检测综合诊断能提高诊断准确率。     

血清首氨酰脯氨酸二肽氨基肽酶同工酶对原发性肝癌的诊断价值

目的研究血清甘氨酰脯氨酸二肽氨基肽酶（ｇｌｙｃｙｌｐｒｏｌｉｎｅ　ｄｉｐｅｐｔｉｄｙｌ　ａｍｉｎｏｐｅｐｔｉｄａｓｅ,ＧＰＤＡ）同工酶对ＰＨＣ的诊断价值,提高早期 ＰＨＣ和 ＡＦＰ阴性ＰＨＣ诊断水平。方法采用阶段梯度垂直平板聚丙烯酰胺凝胶电泳法进行血清ＧＰＤＡ同工酶分离检测,同步测定比较ＰＨＣ患者血清ＧＰＤＡ同工酶与ＧＰＤＡ,Ｑ．活性（ＴＧＰＤ）、ＡＦＰ、肿瘤大小及ＡＬＴ的关系。结果血清ＧＰＤＡ按其泳动速度可分为快带（ＧＰＤＡ－Ｆ）与慢带（ＧＰＤＡ－Ｓ）两种同工酶。ＰＨＣ患者血清ＧＰＤＡ－Ｆ的阳性率为８５．３％,而肝外肿瘤、转移性肝癌、肝炎后肝硬化、慢性肝炎中阳性率均较低,正常对照组以及良性肝占位全部阴性。ＧＰＤＡ－Ｆ阳性率在ＴＧＰＤＡ活性增高组明显高于正常组（９４．４％对７５０％）,ＧＰＤＡ－Ｆ与ＡＦＰ及肿瘤大小无关系。良性肝病ＡＬＴ升高组ＧＰＤＡ－Ｆ阳性率高于ＡＬＴ正常组,而ＰＨＣ患者ＧＰＤＡ－Ｆ与ＡＬＴ无关。ＰＨＣ患者ＧＰＤＡ－Ｆ呈持续阳性,良性肝病患者ＧＰＤＡ－Ｆ常为一过性阳性。结论ＧＰＤＡ－Ｆ为一新的ＰＨＣ血清标志物,有助于ＰＨＣ的定性诊断,尤其对早期ＰＨＣ及ＡＦＰ阴性ＰＨＣ的诊断具有重要价值。     

The possibility of determining N-acetyl-beta-D-glucosaminidase isoenzymes under alkaline conditions

OBJECTIVES: To have a reliable diagnostic test, the influence of urine pH on the determination of the total activity of N-acetyl-beta-D-glucosaminidase (NAG) and NAG isoenzyme activities was studied. DESIGN AND METHODS: After ultrafiltration and dialysis of the acidic and alkaline urines, the B, A, and A2 forms of NAG were separated by ion-exchange chromatography on DEAE cellulose. RESULTS: A significant decrease in the total activity of NAG in alkaline urines (pH around 8 or higher) was found, which makes this determination unreliable. Analysis of the isoenzymic profiles obtained for weakly acidic and alkaline urines (in the pH range from 5.5. to 10.0) showed that the percent fractions of the individual isoenzyme activities in the total NAG activity and their ratios changed only at pH values above 9.5. CONCLUSIONS: The determination of the denoted isoenzymes of urinary NAG after ultrafiltration, dialysis, and chromatographic separation on DEAE cellulose is reliable in a wide range of alkaline pH values of urine.     

Isoenzyme studies in human leukemia-lymphoma cell lines--1. Carboxylic esterase

The isoenzyme patterns of carboxylic esterase (E.C. 3.1.1.1) were studied in 74 proven human leukemia-lymphoma and 12 normal B-lymphoblastoid cell lines. These cell lines have been extensively phenotyped using poly- and monoclonal antibodies. Esterase isoenzymes were separated by isoelectric focusing and visualized by histo-cytochemical techniques. No leukemia-specific or (except for monocytes) blood cell type-specific isoenzyme or isoenzyme pattern could be detected. The monocytic element in some cell lines was characterized by a strong isoenzyme band which could be selectively and completely inhibited by sodium fluoride. The enzyme phenotypes were stably expressed in all subcultures of a given cell line and did not appear to have any cell cycle dependency. The leukemia-lymphoma cell lines have been subclassified into four major groups according to immunological parameters: T-cell, B-cell, myelomonocytic and non-T, non-B-cell. On the basis of immunological data the T-cell lines were assigned to five stages of differentiation. The number and staining intensity of the isoenzymes increased with differentiation of the T-cells paralleling the expression of immunological markers. The B-cell leukemia-lymphoma cell lines were divided into pre B-, B-, Burkitt lymphoma, multiple myeloma and hairy cell leukemia cell lines. Substantial variability among the isoenzyme patterns was detected ranging from immature profiles of pre B-cell lines to complete isoenzyme repertoires of multiple myeloma cell lines. No significant difference was seen between the isoenzymes of mature B-cell lines and normal B-lymphoblastoid cell lines. The most prominent feature seen in myelomonocytic cell lines was the monocytic band indicating a monocytic origin and separating the 'monocytoid' from the 'pure myeloid' cell lines. Considerable heterogeneity in the isoenzyme patterns was observed in the non-T, non-B cell groups which comprised erythroleukemia cell lines and cell lines arrested at a very early stage of lymphoid differentiation. These latter cell lines together with some T- and B-cell lines shared the common characteristics of positivity for cALLA, TdT and Ia antigens and an immature, incomplete isoenzyme profile. The results support the notions of maturation arrest and normal gene expression in leukemic cell populations. Furthermore, the importance of biochemical studies as part of the multiple marker analysis could be demonstrated.     

Isoenzyme studies in human leukemia-lymphoma cell lines--IV. Lactate dehydrogenase

Isoelectric focusing (IEF) in horizontal polyacrylamide gels has been used to separate lactate dehydrogenase (LDH) isoenzymes in 97 human permanent hematopoietic cell lines (85 leukemia-lymphoma cell lines and 12 'normal' B-lymphoblastoid cell lines). Maximally 8 LDH bands were seen; the electrophoretically detectable bands 4 and 5 could be separated by IEF into 2 and 3 isoenzymes, respectively. The LDH patterns have been found to vary both in number of isoenzymes and in relative intensity in different cell lines depending upon the stage at which arrest of differentiation occurred. These differences can be used to analyse and distinguish different cell lines. The method should provide a valuable supplement to the enzymatic phenotyping and complete characterization of fresh and cultured leukemias and for the monitoring of phenotypic changes occurring during induction of differentiation.     

Isoenzyme der Enolase in Erythrocyten Neugeborener und Erwachsener

Zusammenfassung Durch Elektrophorese auf Cellulose-Acetat-Folie lassen sich in menschlichen erythrocyten 3 Isoenzyme der Enolase (E.C. 4.2.1.11) trenen. Die in Erythrocyten Neugeborener gegenüber Erwachsenen stark erhöhte Gesamtaktivität der Enolase beruht überwiegend auf einer Vermehrung des Isoenzyms I. Die Isoenzymverteilung der Erythrocyten-Enolase ist vom Zellalter unabhängig.

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Enolase isoenzymes in erythrocytes of neonates and adults

A method is described for the separation of human erythrocyte enolase (E.C. 4.2.1.11) isoenzymes by electrophoresis on cellulose acetate foils (Cellogel). 3 isoenzymes were found in erythrocytes of neonates and of adults. The higher total activity of enolase in the red cells of neonates compared with adults is mainly due to an increase of the isoenzyme type I. The isoenzyme pattern of erythrocyte enolase is not influenced by the age of the cells.

Mit Untertützung durch die Deutsche Forschungsgemeinschaft (Ba 246/8-9).