Nagoya Journal of Medical Science

Effect of fixatives on the unstable functional states of mitochondria was studied. The energized configuration of the cristal membrane of beef heart mitochondria can be maintained and visualized by the electron microscope if fixatives are introduced before the oxygen supply is exhausted. Since the exhaustion of the available oxygen supply is completed in 5-20 seconds, it is impossible to apply the method of sedimenting the mitochondria prior to fixation for studying the energized configurational states of mitochondria. The direct addition of glutaraldehyde followed by osmium tetroxide to the mitochondrial suspension is most effective for freezing the configurational state of cristal membrane. Fixation with glutaraldehyde appears to be complete within 1-2 seconds even at 0°C. Osmium tetroxide alone can also "freeze" the energized configuration by fixation but the concentration of the fixative is critical. The problem of capturing the configurational state applies not only to energized transitions (nonenergized to energized) but also to nonenergized transitions (orthodox to aggregated). When the level of glutaraldehyde or osmium tetroxide is respectively too low or too high, the mitochondria will undergo a transition from the aggregated to the orthodox configuration before fixation is complete. Light-scattering measurements which provide an independent method for monitoring configurational changes in mitochondria confirm that the conditions for fixation which lead to stabilization of the energized state as judged by electron microscopy, also show maintenance of configuration.

Oral galactose tolerance tests were performed on 56 patients with liver disease, 31 with diabetes mellitus and 17 normal subjects. Intravenous galactose tolerance tests were done on 25 liver disease, 6 diabetes mellitus and 6 normal subjects. Each liver disease was histologically comfirmed by needle biopsy under laparoscopy. Serum galactose and glucose were measured gaschromatographically. Oral galactose tolerance test is useful for the diagnosis of liver cirrhosis and chronic hepatitis, but metabolic disorder of galactose can be found in diabetics. Therefore abnormalities in oral galactose tolerance test do not necessarily mean the existence of hepatic damage. There exists a relatively good correlation between the degree of abnormal oral galactose tolerance test and the level of serum r-globulin, cholinesterase and ICG disappearance rate. As for intravenous galactose tolerance test, half life is a useful index as well as oral glaactose tolerance test. But it can be concluded that oral galactose tolerance test is superior to intravenous galactose tolerance test from the point of view of technical simplicity and the burden to patients.

Light, electron microscopic, enzyme histochemical and ultracytochemical studies were performed on specimens of oncocytoma, obtained from an asympto- matic mass in the right parotid gland of a 61 year old Japanese female.
With the light microscope, the diagnosis was benign oncocytoma. Electron microscopically, the cytoplasm was filled with a vast number of mitochondria, with numerous cristae, some of which contained glycogen particles.
The tumor cells showed a strong activity of histochemically demonstrable cytochrome oxidase.
At ultrastructural level for the demonstration of cytochrome oxidase, using DAB, many of the mitochondria had reaction product, which was restricted only to the cristae and the intracristal space. Also, the glycogen-containing-mitochondria had the enzyme activity on the cristae and in the intracristal space.
The mitochondria of oncocytes could have considerably active function.
These results suggest that oncocytoma which is characterized by numerous mitochondria could be in hyperactive function rather than degeneration.