The present study revealed that the intravital reduction of Nitro-BT can occur in the absence of any exogenous substrate in the living hair cells of the cochlea in nembutal anaesthetized guinea pigs. Electron transfer inhibitors, such as amytal and malonate considerably impeded the intravital reduction of Nitro-BT. The reduction of Nitro-BT was completely blocked after heating at 100°C for 15 minutes and cooling at-20°C for 30 minutes. The reduction of Nitro-BT continued to some extent in the hair cells left at room temperature at least for 60 minutes after sacrification. Acute complete starvation caused considerable decrease in the intravital reduction of Nitro-BT and vitamin B₁ deficiency did rather slightly. The intravital reduction of Nitro-BT seemed to be due to the intravital multi-enzymatic tetrazolium reducing reaction. The present study suggested that the intravital enzymatic tetrazolium reducing reaction may depend on the amount of electrons which are transferred by CoQ in the respiratory chain to Nitro-BT. The amount of electrons transferred by CoQ to Nitro-BT was considered the intravital reduction potency of CoQ. The intravital reduction potency of CoQ seemed to depend on cellular metabolism and its rate in the hair cells.

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Chinoform (7-iodo-5-chloro-8-hydroxyquinoline) which is a derivative of oxin (8-hydroxyquinoline) can chelate with some metals as well as oxin and other derivatives. Applying this principle, the techniques of histochemical demonstration for chinoform, which is probably the causal agent of subacute myelo-optico-neuropathy (SMON), were devised.
Copper, iron and silver were used as substances to be chelated with chinoform and they were demonstrated histochemically. In using these techniques, it was necessary to perform these chemical reactions in Tween 80 solution.
The possibilities of these techniques were examined using various kinds of materials.
Insoluble chinoform hydrolyzed from chinoform glucuronide as a substrate by β-glucuronidase activity deposited ill situ positively and was demonstrated histochemically. In addition, the devised techniques were tested and comparisons were made with the autoradiographic findings in the tissue sections of rabbits to which ¹³¹I-chinoform was administered.

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Azan-Mallory stain of thyroidal colloid and its relation to the functional state of the thyroid were studied by animal experiments and on clinical materials. Thyroidal colloid shows basophilia and acidophilia in elevated and resting state of thyroid function, respectively. The acidophilic colloid itself represents basophilia at its margin adjacent to the follicular cells. In case of adenoma, well differentiated follicles contain acidophilic colloid, and less differentiated, basophilic colloid. In case of adenocarcinoma, both acidophilic and basophilic colloids are irregulary admixed in a single follicle.

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Terminal degeneration of the rat spinal cord after the transection was investigated using the ZIO staining method. Degenerating processes were classified into three types according to ZIO reactivity of mitochondria, synaptic vesicles, synaptic membrane and matrices of degenerating terminals. It was established that ZIO staining was useful not only for the detection of degenerating terminals but also for the determination of degenerating stages.

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In fed yellow KK mice, the pancreatic B cells showed severe degranulation and diminished content of zinc. In these islets, there were observed small number of secretory granules which could be classified into pale granules and dense granules with their electron densities. Both granules underwent emiocytosis. Zinc was detected in cores of dense granules as fine precipitates of silver sulfide. The pancreatic islets from 14 hours fasted mice were heavily granulated but still low in zinc reaction. By electron microscopy, B cells were filled with pale granules with very low zinc contents. Fortyeight hours fasted mice showed granulated and zinc-rich islets. In their B cells, many dense granules could be observed.
These findings suggest conversion of pale granules into dense ones, on which zinc plays some role. Furthermore, it is likely that premature granules undergo secretion without being matured into dense granules in animals with hyperinsulinemia.

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For the purpose of increasing our knowledge concerning the morphologic aspects of the effects of insulin on a chondrogenesis and a micromelia, tibiotarsi of chick embryos injected with a single dose of 4 units of insulin into yolk sac on the 5th day of incubation were studied by the light microscope using histochemical methods. Two days after insulin treatment, the anlage of the tibiotarsus showed the imperfect differentiation into cartilage. The prechondroblasts failed to accumulate glycogen in the cytoplasm. Although the normal-like chondrogenic activity occured in the perichondrium after 5 days of the injection, the cells in the central areas of the epiphysial plate showed imperfect differentiation into chondroblasts and some of them were degenerated. The degenerative alterations in the central areas of the epiphysial plate 7 days after the treatment became more severe than the previous stage. After 13 days of the injection the degenerated tissues in the central areas were completely replaced by myeloid tissues. It could be deduced from the above that the failure to form epiphysial plate in insulin-treated chick embryos is due to the reduced interstitial growth, thus causing micromelia.

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Submicroscopic effects of insulin on the chondrogenesis and a micromelia were investigated with electron microscopic and histochemical methods, such as glycogen, acid phosphatase, acid mucopolysaccharide. The chondrogenic cells of the tibiotarsal anlage of 7 day chick embryos treated with 4 units of insulin on the 5th day of incubation had poorly differenciated cytoplasmic organelles. Glycogen granules conspicuously decreased in number than the control. After 5 days of the treatment the cells showed somewhat degenerative changes characterized by the degenerated cell organelles and the increase of acid phosphatase activity. Lead phosphate deposits were observed in the cisterna, vesicles of the Colgi areas and the lipid droplets. The glycogen granules totally disappeared from the cytoplasm. Seven days after the treatment the cells were decreased in number. The absence of Golgi vacuoles correlate with suppression of matrix formation. Matrix granules which showed ruthenium red positive reaction, remarkably decreased in number than the previous stages and only traces of them could be seen associated with fibrills.

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