GTP binding protein (G-protein) associated with membrane and involved in signal transduction was isolated from bovine brain, and molecular weight of G protein was observed. As the results, cell membranes were homogenized from bovine brain tissues and proteins of membrane were gained using 1% cholate, and progressed the chromatography. The purification process was performed by step, DEAE-Sephacel, Ulttrogel AcA 34 and heptylamine-Sepharose column chromatography. The chromatographic fractions were confirmed by GTP binding assay and SDS-polyacrylamide gel electrophoresis. Molecular weight of Goa was revealed 39,000 dalton and GR 36,000 dalton. One more step of heptylamine-Sepharose was enforced to purify the GTP binding protein. Finally I gained the GTP binding protein isolated subtype of Goalpha and Gbeta.
Brain* ; Cell Membrane ; Cholates ; Chromatography ; Electrophoresis ; GTP-Binding Proteins ; Guanosine Triphosphate* ; Membranes ; Molecular Weight ; Signal Transduction

No abstract available.
Drug Resistance, Multiple* ; Drug Therapy*

In rat kidney, the changes in concentrations of nucleotides and their derivatives during ischemia induced by renal artery ligation was measured quantitatively with high performance liquid chromatography (HPLC). After the ligation of renal artery for 60minutes, the concentrations of the nucleotides and derivatives to 80.7±18.39 µg (p<0.01); ATP, 307.2±56.68 µg to 47.6±5.95µg (p<0.01); ADP+AMP, 227.1±7.98 µg to 61.4±3.92 µg (P<0.01); NAD+, 217.9±4.49 µg to 126.6±10.44 µg (P<0.01); GTP, 202.5±23.76 µg to 117.7±14.24 µg (P<0.05); GMP, 54.5±9.03µg to 23.7±0.46 µg (p<0.05), and inosine, 16.6±3.45 µg to 7.8±0.87 µg (P<0.05). But hypoxanthine and xanthine were significantly increased from 113.0±15.58µg to 159.7±12.97µg (P<0.05) and from 87.7±6.77µg to 173.1±12.52µg (P<0.01). In ischemic kidney, concentration of ATP was decreased to 39.9% of control at 10 minutes, 19.8% at 30 minutes, and 15.5% at 60 minutes, and ADP+AMP were decreased to 70.3% of control at 10 minutes, 67.3% at 30 minutes, and to 27.0% at 60 minutes, but hypoxanthine and xanthine were increased to 121.5% and 127.1% at 10 minutes, 126.0% and 174.4% at 30 minutes, and 141.4% and 197.3% at 60 minutes. Total adenosine nucleotides were decreased to 20.3% of control during 60 minutes of ischemia, but hypoxanthine and xanthine were increased to 157.5% of control. These results suggest that the changes in the concentration of nucleotides and their metabolic derivatives are useful indices of the extents of tissue ischemia in rat kidney.
Adenosine ; Adenosine Triphosphate ; Animals ; Chromatography, High Pressure Liquid* ; Chromatography, Liquid ; Guanosine Triphosphate ; Hypoxanthine ; Inosine ; Ischemia* ; Kidney ; Ligation ; Nucleotides* ; Rats* ; Renal Artery ; Xanthine

The development of multidrug-resistant tumor cell population is a major problem in the chemotherapy of human cancer. These cells are often cross resistant to unrelated drugs and the precise mechanisms of multidrug resistant phenotype of tumor cells has not been fully elucidated. Cisplatin resistant tumor cell (SNU-1/Cis₅) was induced from human stomach cancer cell line (SNU-1) in vitro. Growth profiles of survival cells were observed during 5 days by thiazolyl blue (MTT) assay. To investigate the cross resistance of various anticancer drugs in SNU-1 and SNU-1/Cis5, We compared the value of IC₅₀-drug concentration at 50% survival of control and gained relative resistances (RR). The RR for SNC-1/Cis₅ were as follows; vinblastine, > 43.0; epirubicin, 22.9; dactinomycin, 16.0; etoposide, 15.0; vincristine, 9.2; adriamycin, 5.7; aclarubicin, 5.3. But 5-fluorouracil, methotrexate, daunorubicin have not cross resistance with cisplatin. Resistant inhibition values of 10µM verapamil for SNU-1/Cis₅ were as follows; vincristine, 13.1; epirubicin, 10.0; etoposide, 6.3; vinblastine, 4.4; dactinomycin, 3.6; daunorubicin, 2.4. Membrane proteins of 51,400 and 81,300 daltons were identified by radioiodination with SDS-PAGE, which might represented the drug resistance.
Aclarubicin ; Cell Line ; Cisplatin* ; Dactinomycin ; Daunorubicin ; Doxorubicin ; Drug Resistance ; Drug Resistance, Multiple* ; Drug Therapy ; Electrophoresis, Polyacrylamide Gel ; Epirubicin ; Etoposide ; Fluorouracil ; Humans* ; In Vitro Techniques ; Membrane Proteins ; Methotrexate ; P-Glycoprotein ; Phenotype ; Stomach Neoplasms* ; Stomach* ; Verapamil* ; Vinblastine ; Vincristine

The one event on signaling mechanism is the cleavage by adenyl cyclase of ATP into second messenger, cyclic AMP. The other transfer system of inositol metabolism, it is widely recognized that hydrolysis of the minor membrane lipid phosphoinositide bisphosphate (PIP₂) initiated by occupation of certain receptors and catalyzed by phospholipase C, lead to toe generation of the two intracellular messengers, inositol triphosphate (IP₃) and diacylglycerol (DG). IP₃ is converted to inositol tetrakisphosphate (IP₄) by IP₃ kinase. In the present study, it is that purification of calmodulin is used by phenyl-Sepharose CL-4B chromatography, it's molecular weigh, 17,000 in SDS-polyacrylamide gel electrophoresis. In order to observe the affinity between calmodulin (CaM)-Affigel 15 and IP₃ kinase, and isolated IP₃ kinase, was applied in CaM-Affigel with Ca²⁺ equilibrium buffer and EGTA equilibrium buffer. We compared with binding and elution effect of IP₃ kinase in several condition of buffer. In affinity of binding, Ca²⁺ equilibrium buffer was in the most proper condition, and elution, CaM/Ca²⁺buffer (CE 1 10.36, CE2 12.76pM/min/mg of protein) was effected much more than EGTA buffer (E2 1.48, E 2.43pM/min/mg of protein), but CaM/Ca²⁺stimulate the activity of IP₃ kinase. And then, several detergents such as sodium deoxycholate, tween 20, cholic acid, polyethylene glycol, chaps were applied. The 0.2% chaps buffer (E2 23.19, E3 8.05pnM/min/mg of protein) was the most effective in elution of IP3 kinase.
Adenosine Triphosphate ; Adenylyl Cyclases ; Brain* ; Calmodulin ; Cholic Acid ; Chromatography ; Cyclic AMP ; Deoxycholic Acid ; Detergents ; Egtazic Acid ; Electrophoresis ; Hydrolysis ; Inositol* ; Membranes ; Metabolism ; Occupations ; Phosphotransferases* ; Polyethylene Glycols ; Polysorbates ; Second Messenger Systems ; Toes ; Type C Phospholipases

PURPOSE: Growth hormone(GH) has not only growth promoting effect but also various metabolic effects. We evaluated GH effects by anthrometric data, biochmical data, electrolytes and simple CT in patients with Prader-Willi syndrome. METHODS: Nine children with Prader-Willi syndrome(PWS) were studied. The children were treated with GH(0.6U/kg/week) for 6 months. Before and after therapy we measured height, weight, waist, hip, and thigh. Blood sampling for eletrolytes, HgA1C, lipid profiles and other biochemistry were done in all patients before and after therapy. We also compared fat distribution with scan. RESULTS: Height standard deviation (SD) score increased from -0.7 to -0.5 and weight SD score decreased from 5.3 to 4.9. Body mass index(BMI) decreased from 28.2kg/m2 to 27.2kg/m2. But the changes in height, weight and BMI were not significant statistically. The waist/hip ratio decreased from 1.04 to 0.97(P<0.05), Thigh circumference had been decreased from 58.2+/-21.7cm to 49.9+/-6.9cm insignificantly. The visceral fat were decreased from 7,613+/-1,760 to 5,022+/-1,533 after GH therapy, and thigh muscle mass was increased from 6,358+/-1,616 to 7,175+/-2,155 (P<0.05). Total cholesterol and triglyceride decreased and HDL cholesterol increased after therapy although they were insignificant statistically. There were no differences in electrolytes, HgA1C, other biochemistry(Ca, P, protein, albumin, BUN, Cr) before and after therapy. CONCLUSION: In children with PWS, waist/hip ratio and fat mass were reduced and muscle mass was increased after GH therapy. There was tendency that total cholesterol and triglyceride decreased and HDL cholesterol increased after therapy. We confirmed that GH therapy had not only growth promoting effect but also metabolic effect on lipid and protein metabolism in children with PWS.
Biochemistry ; Child ; Cholesterol ; Cholesterol, HDL ; Electrolytes ; Growth Hormone* ; Hip ; Humans ; Intra-Abdominal Fat ; Metabolism ; Prader-Willi Syndrome* ; Thigh ; Triglycerides

Glutathione(GSH) has a very important role in detoxification of cells and is closely related to antitumor drug-resistance of cancer cells. In order to evaluate the importance of glutathione metabolism in the drug-resistant cancer cells, the concentration of celluar GSH and activities of y-glutamylcysteine synthetase(GCS), y-glutamyl transpeptidase (GGT) and glutathione S-transferases(GST) in the adriamycin, vincristine, or cisplatin resistant L1210 (L1210AdR; L1210VcR, or L12100s) sublines were measured. Expression and amplification of GCS, GGT, and GST-i7 genes were also observed in the parent L1210 and the drug-resistant L1210 sublines. The concentration of GSH was increased 5.34 fold in L12100s, 2.83 fold in L1210VcR, and 1.78 fo-d in L1210AdR, compared to L1210. The activities of GCS and GGT were -increased in drug-resistant L1210 sublines. The GST activity was increased in L1210VcR and L1210Cis but decreased in L1210AdR compared to L1210. Expression of GCS, GGT, and GST-rr genes were increased in the resistant L1210 sublines compare to the parent L1210 in northern blot analyses. Overexpression of GCS, GGT, and GST-77 were observed in the resistant sublines, and the increases of the concentration of glutathione and the activities of GCS and GGT in the resistant sublines may be involved in a part of the drug-resistance in the resistant sublines.
Blotting, Northern ; Cisplatin ; Doxorubicin ; Drug Resistance ; Glutathione* ; Humans ; Metabolism* ; Parents ; Vincristine

Leiomyomas are the most common benign tumors of the uterus, and although they can occur at any site in the genitourinary tract, they very rarely originate from the urinary bladder. We report a case in which a leiomyoma occurred at that site, deseribing the US, CT, and MRI findings.
Leiomyoma* ; Magnetic Resonance Imaging* ; Urinary Bladder* ; Uterus

Cytochrome P-450 (CP-450) is one of the three components of the liver microsomal enzyme system which hydroxylates fatty acids, hydrocarbons and a variety of drugs and other foreign compounds. Female Balb/c mice were immunized with purified polychlorinated biphenyl (PCB) – induced CP-450 LMII. The spleen cells derived from immunized mice were fused with SP2 myeloma cells using polyethylene glycol (PEG 3500). The hybrid cells were selected by hypoxanthine-aminopterin-thymidine (HAT) medium and the culture fluid were screened by enzyme-linked immunosorbent assay to CP450 LMII. The hybrid cells(×107) were inoculated into intraperitoneal cavity of Balb/c mice for the purpose of production of ascetic fluids. Monoclonal antibody (Mab) was purified from ascitic fluid by DEAE cellulose ion exchange chromatography and I¹²⁵ labeled Mab was also confirmed by autoradiography and SDS-polyacrylamide gel electrophoresis (MW:55,000 and 110,000)
Animals ; Ascitic Fluid ; Autoradiography ; Chromatography, Ion Exchange ; Cytochrome P-450 Enzyme System* ; Cytochromes* ; DEAE-Cellulose ; Electrophoresis ; Enzyme-Linked Immunosorbent Assay ; Fatty Acids ; Female ; Humans ; Hybrid Cells ; Hydrocarbons ; Liver* ; Mice ; Polyethylene Glycols ; Rats* ; Spleen

Band 3, the predominant 95,000 dalton anion transport protein, is the major intrinsic glycoprotein of the human erythrocyte membrane. This anion carrier exists as a dimer and binds the cytoskeletons such as spectrin, ankyrin and actin. And the liposomes are vesicular structures which form spontaneously upon hydration of phospholipids. These artificial lipid vesicles have been investigated as model of the biological membranes and as a mean of improving the delivery of nucleic acids, drugs, proteins and biological substances to specific target tissues and cells. In this study, we were purified Band 3 from the human erythrocyte membrane (ghost) was prepared by hemolysis of intact human erythrocyte with weak alkali-hypotonic solution. Band 6 was removed from ghost by extracting with solution of an ionic strength of 0.15. Band 3 and Band 4 were solubilized selectively by extracting Band 6-depleted ghosts with Triton X-100 under nondenaturing conditions. Band 3 was then purified from Triton X-100 extract treated with p-chloromercuribenzoate by sucrose density gradient ultracentrifugation. This purified Band 3 was incorporated into liposomes prepared by reverse-phase evaporation. Phosphatidyl L-serine and cholesterol (1:1 molar ratio) were dissolved in chloroform and the chloroform was removed by rotator evaporation under reduced pressure. Band 3 solution without Triton X-100 was introduced into a mixture of lipids and diethylether. Diethylether was subsequently removed by evaporation. This purified Band 3 and its incorporation into liposomes were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Actins ; Ankyrins ; Chloroform ; Cholesterol ; Cytoskeleton ; Electrophoresis ; Erythrocyte Membrane* ; Erythrocytes* ; Glycoproteins ; Hemolysis ; Humans* ; Liposomes* ; Membranes ; Molar ; Nucleic Acids ; Octoxynol ; Osmolar Concentration ; Phospholipids ; Serine ; Sodium ; Spectrin ; Sucrose ; Ultracentrifugation