Ultracytochemistry of pancreatic damage induced by excess lysine

The ultracytochemical changes induced in the pancreas by a single large dose of lysine (400 mg/100 g body weight) were studied in male Wistar rats of 7 weeks old. The first changes in the acinar cells were marked swelling of mitochondria with increase in their calcium content and decrease in their ATP content. Early calcium deposits seemed to occur in the matrices of swollen mitochondria and later various patterns occurred. These findings suggested that damage of the acinar cells by excess lysine resulted in breakdown of the mitochondrial membrane barrier to calcium as a very early abnormality, and that extracellular calcium then entered the mitochondrial matrices and inhibited mitochondrial function. Subsequently focal areas of the cytoplasm were degraded. Autophagic vacuoles appeared in these areas, and then acid phosphatase activity in their periphery as a result of fusion with lysosomes. The reaction of acid phosphatase was demonstrated in the locally degraded rough endoplasmic reticulum within or around autophagic vacuoles, suggesting that the endoplasmic reticulum as well as lysosomes participated in the intracellular degradation of cytoplasmic organelles in damaged acinar cells.     

Rapid and simple method for the determination of α1-acid glycoprotein in serum by column liquid chromatography

A rapid and simple method for the determination of α₁-acid glycoprotein (AAG) in serum was developed by using an anion-exchange column for clean-up of serum and a hydroxyapatite column for high-performance liquid chromatography (HPLC). A good correlation was observed between this HPLC method and the conventional radial immunodiffusion method. The method may also be used to determine the AAG concentration in the serum of experimental animals.     

Purification method for α-1-acid glycoprotein with subsequent high-performance liquid chromatographic determination of monosaccharides in plasma of healthy subjects and patients with renal insufficiency

A simple purification method for human plasma α-1-acid glycoprotein (AAG) using an ion-exchange and hydroxyapatite column was developed. The recovery of the method was found to be high. We also improved a determination method for N-acetylneuraminic acid and monosaccharides in the carbohydrate moiety of AAG by using an ion-exchange column and pulse-amperometric detection. By this method, a composition analysis of the carbohydrate moiety of AAG (N-acetylneuraminic acid, fucose, N-acetyl glucosamine, galactose and mannose) was possible with 1.0 ml of plasma. We compared these carbohydrate concentrations in the AAG of patients with renal insufficiency with those of healthy subjects. In the AAG of the patients, the concentrations of N-acetylglucosamine, galactose and mannose were significantly higher than those in the AAG of the healthy subjects.