In this study, we measured the activity of the erythrocyte pyrimidine 5'-nucleotidase(P5N) for 83 workers exposed to lead in their workplace, and analyzed the correlation of the activity with other biological exposure indices of lead. The measurement was performed by using a high performance liquid chromatography (HPLC) with a reverse phased ODS column. The results are as follows; 1. The correlation of the erythrocyte P5N activity with the concentration of blood lead(PbB) was shown to be statistically significant (r=-0.71, p=0.0001). 2. For a group of subjects whose PbB was less than 10microgram/dl, the erythrocyte P5N activity was 14.9+/-1.5 micromole uridine/h/g Hb. 3. For medical surveillance, this study suggests the erythrocyte P5N activity of 12 micromole uridine/h/g Hb is equivalent to 40 microgram/dl of PbB. 4. The correlation coefficients of the erythrocyte P5N activity with other biological exposure indices of lead such as PbB, ZPP, ALA-U, PBU, CP-U, ALAD, and log ALAD were -0.71, -0.64, -0.57, -0.51, -0.50, 0.46 and 0.64, respectively. 5. The correlation coefficients of the PbB with other biological exposure indices of lead for ALAD, P5N, ZPP, PBU, CP-U, and ALA-U were -0.76, -0.71, 0.68, 0.59, 0.42, and 0.41, respectively. The erythrocyte P5N activity can be used as a reliable biological exposure index of lead.
5'-Nucleotidase* ; Chromatography, Liquid ; Erythrocytes*

In this study, we measured the activity of the erythrocyte pyrimidine 5'-nucleotidase (P5N) and urinary N-acetyl-beta-D-glucosaminidase (NAG) from 154 workers exposed to lead and 43 workers not exposed. We analyzed the correlation of the P5N activity and NAG activity with other biological exposure indices of lead such as blood lead (PbB) and zinc protoporphyrin (ZPP). The measurement was performed by using high performance liquid chromatography (HPLC), spectrophotometer and atomic absorption spectrophotometer. The results are as follows: 1. The mean value of P5N activity for workers exposed to lead was 9.50+/-.13 micromol uridine/hr/g Hb and 11.60+/-.2 micromol uridine/hr/g Hb for workers not exported. The P5N activity showed a normal distribution, but the other indices of lead showed logarithmic normal distributions. 2. The P5N activity and ZPP were decreased as PbB wag increased. But the NAG activity had no correlation with changes of PbB. 3. The correlation coefficients of the P5N activity with other biological exposure indices of lead such as PbB, ZPP, NAG activity were -0.72, -0.55, and 0.05, respectively. We speculated that the P5N activity can be used as a reliable biological exposure index of lead but NAG activity can be used as a biological management index of lead.
5'-Nucleotidase* ; Absorption ; Acetylglucosaminidase* ; Chromatography, Liquid ; Erythrocytes* ; Zinc

This experiment was designed to investigate the histochemical and ultrastructural changes of the proximal tubules and glomeruli of rabbits treated with cyclosporin A (CsA) or cyclosporin A and concomitant administration of prednisolone (PD). Rabbits were given with CsA for 1 week (CsAlW) and for 2 weeks (CsA2W) and CsA plus PD for 1 week (CsA +PDIW) and for 2 weeks (CsA+PD2W). The dose of CsA was 5mg/kg/day i.v. and PD was 1 mg/kg/day i.m. At 1 week and 2 weeks after the treatments, the following items were studied of alkaline phosphatase (ALPase). peroxidase, 5'-nucleotidase (SNU) with the ultrastructural enzymatic changes and electron microscopic changes. CsAlW, CsA2W and CsA +PD2W groups showed a decrease of ALPase activity and CsA+PD2W and CsA2W groups showed weakly increased peroxidase activity CsA2W and CsA+PD2W showed a decrease of 5NU activity. Ultrastructural changes of CsAlW CsA2W and CsA+PD2W groups revealed swollen and disrupted mitochondria, loss of brush border in the proximal tubules and thickening of basement membranes, edematous podocytes and fusion of root processes of podocytes in the glomeruli showing the early stage of nephrotoxicity. The concomitant treatment of PD with CsA eased the nephrotoxicity of CsA within 1 week after the treatment.
5'-Nucleotidase ; Alkaline Phosphatase ; Basement Membrane ; Cyclosporine* ; Kidney* ; Microvilli ; Mitochondria ; Peroxidase ; Podocytes ; Prednisolone* ; Rabbits

For the purpose of determining the reference value of erythrocyte pyrimidine 5'-nucleotidase (P5N) activity as a biological indicator to lead exposure, this study was conducted on the total of 225 healthy men who had not been exposed to lead occupationally, in July, 1994. The parameters selected in this study were age, hemoglobin, hematocrit, number of red blood cell, mean corpuscular hemoglobin (MCH), mean corpuscular herloglobin concentration (MCHC), blood lead, and erythrocyte P5N activity. The blood lead concentrations were measured using a flameless atomic absorption spectrophotometer, and erythrocyte P5N activities by Sakai's simple method using a HPLC(1986). The results were obtaina as follows; 1. The distribution of blood lead concentrations revealed log-normal distribution, and geometric mean and standard deviation of blood lead were 4.09 microgram/dl and 1.55 microgram/dl, respectively. 2. The erythrocyte P5N activity showed normal distribution, and the mean and standard deviation of the erythroeyte P5N activity were 12.34 umole uridine/h/g Hb and 2.21 umole uridine/h/g Hb, respectively. 3. All of the selected variables including blood lead concentration did not affect the erythrocyte P5N activity. Although the erythrocyte P5N activities were negatively associated with blood lead level, the correlation coefficient was not statistically significant. 4. From the result of this study, 8.7 micromole uridine/h/g Hb was obtained as a reference value of erythrocyte pyrimidine 5'-nucleotidase activity for the healthy adult male who had not been exposed to lead occupationally.
5'-Nucleotidase ; Absorption ; Adult ; Erythrocyte Indices ; Erythrocytes* ; Hematocrit ; Humans ; Male ; Occupations ; Reference Values*

Sakai's method has been known as the simplest one for determination of erythrocyte pyrimidine 5'-nucleotidase (P5N) activity using high performance liquid chromatography(HPLC). However the drawback of the method is that it is difficult to wash the erythrocyte for isolation. To search for the simpler method, we compared Sakai's method with other methods using whole blood treated with heparin and concanavalin A or whole blood treated with EDTA-2K instead of washing the erythrocyte. The mean concentrations of lead in blood samples collected from 44 male and 16 female workers who are healthy without any exposure to lead in their workplace were 4.30 +/- 1.31 microgram /dl (mean +/-standard deviation), which were measured by frameless atomic absorption spectrophotometer. Erythrocyte P5N activities were measured by 3 methods; Sakai's method(Method I), using whole blood treated with heparin and concanavalin A (Method II), and using whole blood treated with EDTA-2K (Method III). The results were obtained as follows ; 1. The mean of erythrocyte P5N activity by Sakai's method(Method I) were 12.7 +/-2.47 amole uridine/hr/gm of Hb. 2. The mean of erythrocyte P5N activity by the method using heparinized whole blood treated with concanavalin A(Method II) were 13.1 +/-2.41 micromole uridine/hr/gm of Hb. 3. The difference of mean erythrocyte P5N activity between Method I and Method was not significant. 4. The erythrocyte P5N activity by the method using whole blood treated with EDTA-2K (Method III) was significantly different from Method I. We thought that omission of incubation period which was required on Method III using EDTA-2K caused the difference between Method I and Method III. 5. Simple linear regression equation for erythrocyte P5N activity between Method I (Y) and Method II(X) was significant: Y = -0.012 + 0.9724 X. These results suggest that the method using whole blood treated with heparin and concanavalin A is simpler to examine the erythrocyte P5N activity as a biological indicator of lead intoxication than Sakai's method.
5'-Nucleotidase* ; Absorption ; Concanavalin A ; Erythrocytes* ; Female ; Heparin ; Humans ; Linear Models ; Male

Sakai's method has been known as the simplest one for determination of erythrocyte pyrimidine 5'-nucleotidase (P5N) activity using high performance liquid chromatography(HPLC). However the drawback of the method is that it is difficult to wash the erythrocyte for isolation. To search for the simpler method, we compared Sakai's method with other methods using whole blood treated with heparin and concanavalin A or whole blood treated with EDTA-2K instead of washing the erythrocyte. The mean concentrations of lead in blood samples collected from 44 male and 16 female workers who are healthy without any exposure to lead in their workplace were 4.30 +/- 1.31 microgram /dl (mean +/-standard deviation), which were measured by frameless atomic absorption spectrophotometer. Erythrocyte P5N activities were measured by 3 methods; Sakai's method(Method I), using whole blood treated with heparin and concanavalin A (Method II), and using whole blood treated with EDTA-2K (Method III). The results were obtained as follows ; 1. The mean of erythrocyte P5N activity by Sakai's method(Method I) were 12.7 +/-2.47 amole uridine/hr/gm of Hb. 2. The mean of erythrocyte P5N activity by the method using heparinized whole blood treated with concanavalin A(Method II) were 13.1 +/-2.41 micromole uridine/hr/gm of Hb. 3. The difference of mean erythrocyte P5N activity between Method I and Method was not significant. 4. The erythrocyte P5N activity by the method using whole blood treated with EDTA-2K (Method III) was significantly different from Method I. We thought that omission of incubation period which was required on Method III using EDTA-2K caused the difference between Method I and Method III. 5. Simple linear regression equation for erythrocyte P5N activity between Method I (Y) and Method II(X) was significant: Y = -0.012 + 0.9724 X. These results suggest that the method using whole blood treated with heparin and concanavalin A is simpler to examine the erythrocyte P5N activity as a biological indicator of lead intoxication than Sakai's method.
5'-Nucleotidase* ; Absorption ; Concanavalin A ; Erythrocytes* ; Female ; Heparin ; Humans ; Linear Models ; Male

OBJECTIVE: Lithium inhibits the action of inositol monophosphatase(IMPase) in phosphoinositide(PI) signal transduction system at therapeutically relevant concentration. The depletion of inositol by lithium itself cannt explain the lithium's therapeutic effect. However, attention has focused on the abnormality of PI signal transduction system as the pathophysiology of bipolar affective disorder(BPD). We investigated whether IMPase mRNA levels of lymphocytes would be different between BPD patients(n=16) and age, sex-matched normal controls(n=16). We also investigated the change of IMPase mRNA level by lithium during 4 weeks to probe the possibility that IMPase mRNA levels could predict the therapeutic response to lithium and clinical course. METHOD: Relative IMPase mRNA levels in lymphocyte were quantified by reverse transcriptase(RT)-PCR in sixteen drug-free BPD patients and sex, age-matched normal controls. The psychopathology of patients were measured using YMRS(Young Mania Rating Scale) and CGI(Clinical Global Impression). RESULTS: There was no significant difference in IMPase mRNA levels between BPD patients and normal controls. And the IMPase mRNA levels were not significantly changed by 4 week treatment with lithium. However, the basal IMPase mRNA levels were negatively correlated with the changes of CGI after 4 weeks. Furthermore, the patients with relatively high basal IMPase mRNA levels showed much more improvement during 4 weeks. CONCLUSIONS: BPD patients and normal controls were not distinguished by lymphocyte IMPase mRNA level. Although we do not support the hypothesis that lymphocyte IMPase activity would be related with the pathogenesis of BPD and the action of lithium, these data raise the possibility that lymphocyte IMPase mRNA levels could function as a predictor of therapeutic response and clinical course of BPD.
5'-Nucleotidase ; Bipolar Disorder ; Humans ; Inositol* ; Lithium* ; Lymphocytes* ; Mood Disorders* ; Psychopathology ; RNA, Messenger* ; Signal Transduction

The incubation of adenosine 5-monophosphate(AMP) with the homogenates of the epidermis and dermis, which were obtained from the axilIary skin of osmidrosis (bromidrosis) patients, resulted in the formation of adenosine and inorganic phosphate (Pi) without further degradation, as demonstrated by paper chromatography. The conversion of AMP to adenosine in the skin was catalyzed by 5-nucleotidase and alkaline phosphatase. It was found that 5-nucleotidase was present both in the epidermis and dermis, being more active in the latter, and that the enzyme was responsible for more than 80% of the total AMP-hydrolyzing activity present in the skin homogenates. Alkaline phosphatase was shown to be present mainIy in the dermis. And its contribution to AMP hydrolysis was insignificant at pH 7.4. From these results, it is evident that AMP is converted to adenosine chiefly by 5-nucleotidase, which is present in the epidermis and dermis.
5'-Nucleotidase* ; Adenosine ; Adenosine Monophosphate* ; Alkaline Phosphatase ; Chromatography, Paper ; Dermis* ; Epidermis* ; Humans* ; Hydrogen-Ion Concentration ; Hydrolysis* ; Skin*

OBJECTIVETo explore the clinical significance of genetic detection and changes of red cell enzyme activities of pyrimidine 5' nucleotidase (P5'N), pyruvate kinase (PK) and glucose-6-phosphate dehydrogenase (G-6-PD) in patients with α-thalassaemia (α-thal).

METHODSThree α-thal patients were further processed to gene detection by PCR-trans-dot blot and gap-PCR, and red cell enzymes activities by absorbance at 260 and 280 nm (A) for P5'N and fluorescence spot test for PK and G-6-PD.

RESULTSRed cells in 3 α-thal cases were microcytic hypochromic with obvious augmented target cells and basophilic stippling erythrocytes. Two patients had anemia, splenomegaly, hyperbilirubinemia and augmented LDH. HbH was positively identified by hemoglobin electrophoresis and hemoglobin cellulose acetate membrane electrophoresis; the other patient had no such abnormalities. Genotypes of 3 patients were of (-α(3.7)/--(SEA)), (αα(QS)/--(SEA))and (--(SEA)), respectively. The activity of P5'N (but not for PK and G-6-PD) in red cell reduced.

CONCLUSIONSThis is the first documented α-thal with P5'N deficiency. Genetic detection might be clinical significant for the diagnosis and pedigree screening of α-thal.

5'-Nucleotidase ; deficiency ; Adolescent ; Adult ; Erythrocytes ; enzymology ; metabolism ; Female ; Humans ; Male ; Middle Aged ; alpha-Thalassemia ; enzymology ; genetics

This study was purposed to investigate the immunoregulatory effect of endothelial cells derived from mesenchymal stem cells (MSC). The human MSC was induced to differentiate into endothelial cells for one week. The phenotypes were evaluated by flow cytometry, the cell morphologic feature was observed by invert phase-contrast microscope and analysis of capillary formation was performed by using the in vitro angiogenesis kit. The immunoregulatory effect was detected by lymphocyte transformation test. The result indicated that during the differentiation cells grew fast and there was no significant change in the phenotypes, i.e. CD73, CD105, HLA-ABC were positive and CD34, CD80, CD86, HLA-DR, CD31 were negative. Immunofluorescence analysis showed typical expression of the von Willebrand factor. Differentiated MSCs formed capillary-like structure. Endothelial cells derived from MSC also revealed immunosuppressive effect on T cell proliferation in a dose-dependent manner. It is concluded that endothelial cells derived from MSC also harbor immunoregulatory effect on T lymphocytes.
5'-Nucleotidase ; metabolism ; Cell Differentiation ; physiology ; Cells, Cultured ; Child ; Endothelial Cells ; cytology ; immunology ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; T-Lymphocytes ; immunology ; von Willebrand Factor ; metabolism