[Living donor liver transplantation in Kyushu University].

We performed living donor liver transplantation (LDLT) for 40 patients at Kyushu University Hospital, Fukuoka Japan during the period from October 1996 to April 2000. The patients consisted of 32 adults and 8 children with a mean age of 35.8 years (range: 1 year and 10 months to 65 years old). The underlying liver diseases of the 40 patients included the fulminant hepatic failure (n = 14), biliary atresia (n = 7), liver cirrhosis (HCV) (n = 6), primary biliary cirrhosis (n = 5), primary sclerosing cholangitis (n = 2), familiar amyloidotic polyneuropathy (n = 2), Alagille syndrome (n = 1), glycogen storage disease (n = 1), huge hepatic hemangiomas (n = 1), and Wilson's disease (n = 1). All liver grafts were obtained from each patient's family members except for one domino transplant donor's case, comprised of 13 parents, 13 sons and daughters, 11 brothers and sisters, and 3 wives. The donors are presently all doing well. The patient survival rate is presently 92.5%.

Influence of blood proteins on biomedical analysis. XII. Effects of glycation on gliclazide (oral hypoglycemic drug)-binding with serum albumin in diabetics.

The sera of diabetic patients showed an inverse correlation (r = -0.67, n = 57) between free gliclazide (oral hypoglycemic drug) level and the fructosamine value. The binding capacity of the primary binding site for gliclazide in the albumin molecule was increased from 4.5 x 10(-4) to 8.0 x 10(-4) M-1 by glycation of albumin, but not that of the secondary binding site (1.2 x 10(-4) M-1). This suggests that the glycation of albumin increases its total binding capacity for gliclazide, resulting in a low free gliclazide level. Therefore, a low hypoglycemic activity of the drug is observed when it is administered to diabetic patients with hyperglycemia.

Correlation between glycated lipoproteins and fructosamine level in serum.

The correlation between the level of fructosamine and glycated proteins, including glycated lipoproteins, in serum from diabetic and nondiabetic subjects was studied. Assay of glycated proteins in serum was performed using an agarose gel film electrophoresis with nitroblue tetrazolium coloration. Glycated albumin correlated well with the fructosamine level in the diabetics (r = 0.83-0.92, p less than 0.01) but showed no correlation with the nondiabetics (r = 0.25-0.26). Also, a high correlation between the glycated beta-lipoprotein and fructosamine levels was observed in diabetic patients with hyperglycemia and in nondiabetic subjects with a high risk of atherogenesis (atherogenic index, low-density lipoprotein-cholesterol/high-density lipoprotein-cholesterol greater than 2.8) (r = 0.51-0.66, p less than 0.01). Nondiabetics with a high level of beta-lipoprotein, which is well known to cause high atherogenesity, showed a high level of glycated beta-lipoprotein similar to that in the diabetic groups with hyperglycemia; therefore, the high level of glycated beta-lipoprotein seems to be attributable not only to the hyperglycemia-accelerated glycation of beta-lipoprotein but also to an increase in the level of beta-lipoprotein in serum. Consequently, the present results show that the fructosamine level in serum reflects not only the glycation of albumin but also that of lipoproteins which are known to increase in diabetes mellitus.

Agarose gel electrophoresis with nitroblue tetrazolium coloration for determination of glycated lipoproteins in serum from diabetics.

We describe a simple method for determining glycated lipoproteins (glc LPs) in serum by agarose gel film electrophoresis, with color development with nitroblue tetrazolium. The resulting blue bands on the film were measured densitometrically at 545 nm to quantify alpha-, pre beta-, and beta-glc LPs. Each glc LP concentration (mmol/L) was calculated from the resulting percentage multiplied by the value for serum fructosamine. Only glc beta-LP was significantly correlated with the atherogenic index: low-density LP-cholesterol/high-density LP-cholesterol (r = 0.545, P less than 0.01). The concentration of glc beta-LP in sera from diabetics was 2.2-fold higher (0.84 mmol/L) than that (0.38 mmol/L) in normal individuals. Diabetic patients with complications had higher concentrations of glc beta-LP, with large individual variations, than did patients without complications, the greatest concentration (1.02 mmol/L) being found in patients with diabetic retinopathy and (or) nephropathy. The concentration of glc beta-LP (glc LDL) in serum seems to depend on the extent and duration of hyperglycemia; it may also be a useful diagnostic indicator of diabetic atherogenesis, microangiopathy (e.g., retinopathy or nephropathy), and other complications.

Influence of blood proteins on biomedical analysis. VIII. Attempts at purification of gliclazide-degrading factor in human serum.

Gliclazide(hypoglycemic drug having sulfonylurea structure)-degrading activity was found in fraction M(macroglobulin, Fr. M) obtained from pooled human serum by gel filtration using a Sephadex G-150 column. The main degrading activity was in the fraction eluted from the Fr. M-subjected DEAE-cellulose column with 0.4 M phosphate buffer (pH 5.2), and the gliclazide-degrading protein localized around alpha 2 to beta-globulin on an electrophoretic pattern using a cellulose acetate membrane. The degrading activity was enhanced about two-fold by lyophilizing Fr. M solution containing a higher sodium phosphate (Na2HPO4-NaH2PO4), over 0.27 M. This indicates that the appearance and enhancement of the degrading activity required the combination of the lyophilization of the sample solution and a certain initial concentration of sodium phosphate prior to lyophilization.

Pharmacokinetics of gliclazide in healthy and diabetic subjects.

The pharmacokinetics of total and free gliclazide, 1-(3-azabicyclo[3,3,0]oct-3-yl)-3-(p-tolylsulfonyl)urea, a potential hypoglycemic drug, was studied in healthy (n = 12) and diabetic (n = 12) subjects. The serum level of gliclazide was determined by a high-performance liquid chromatographic method (HPLC). The free fraction of gliclazide was obtained from serum by an ultrafiltration technique using a collodion membrane. The mean adsorption of gliclazide to the membrane was approximately 50% when the membrane was used more than twice. Therefore, the gliclazide level in the filtrate was corrected by doubling the apparent value. The ratio of gliclazide-protein binding remained constant at approximately 92% in serum after administration to healthy and diabetic subjects. The mean pharmacokinetic parameters of elimination rate (ke), time to reach the peak level (tmax), elimination half-life (t 1/2), and volume of distribution (Vd) were 0.07 h-1, 2.8 h, 12.3 h, and 17.4 L, respectively. The parameters did not differ significantly between healthy and diabetic subjects or between single and successive administrations; moreover, they did not differ between the free and total drug level. Although there were intersubject variations, the therapeutic effects of oral administration of gliclazide on serum glucose and insulin levels were found in four diabetic patients. The results of this study show that the pharmacokinetics of the total gliclazide level reflect those of the free gliclazide in serum.