Fourteen suspected cases of drug-induced hepatitis were diagnostically confirmed by determinating serum enzyme activities following provocative readministration of causative agents. After recovery of their clinical course, the 14 cases were readministrated about half a daily dose of their respective causative agent. Then, the activities of several seru m enzymes were chronologically determined. In 24-48 hours, all cases showed a marked increase in glucose-6-phosphatase, isocitric dehydrogenase and aldolase. Rabbits with experimental liver injury induced by 2-hydroxy-methylene-17 α-methyldihy-drotestosterone showed almost identical results in serum enzyme activities as with those of human cases. Comments are made on the sorts of inducing drugs, incubation periods, symptoms and signs, clinical course, laboratory data, histological findings in liver biopsies, diagnosis, treatment and pathogenesis of drug-induced hepatitis.
It has been regarded that all the histochemical methods so far reported to detect adrenal ketosteroids are of little practical value. The main reason for this invalidity seems to be that the ketosteroid distribution is obscured by the similar positive reaction presented by aliphatic aldehydes that always co-exist with ketosteroids. It is noticeable that formalin appears to increase the atmospheric oxidization effect which will cause or participate in the production of aliphatic aldehydes in formalin fixed tissues as well as frozen sections. Moreover, no suitable carbonyl reaction to distinguish the ketone group of ketosteroids from aliphatic aldehydes in frozen sections, has yet been discovered. In respect to the reducing reaction methods, even when fresh frozen s e ctions are used, the presence of various reducing substances in those sections to seems obscure the relatively faint reducing reaction of ketosteroids. When carbonyl reaction methods are applied to fresh frozen sections, positive results can be hardly obtained, even with the Ashbel-Seligman method indicating the most clear positive color among several carbonyl reaction methods, because it is supposed that these methods are not sensitive enough to detect so small amount of ketosteroid contained in sections. Therefore, the author and others have contrived the iron hematoxylin-sudan III staining method which is regarded to detect ketosteroids indirectly by showing the localization of lipoids or proteins combind with ketosteroids in tissues fixed in dichromate solution containing no formalin. Thus in order to confirm the histochemical validity of this method, we investigated the nature of sudanophilic granules and iron hematoxylinophilic granules found in paraffin sections to which this method is applied. The result was that the sudanophilic granules were found to be a rsudanophilic polymer in paraffin sections where a type of ketosteroid-combining phospholipid was rendered insoluble in fat-solvents through the peculiar oxydizing action of potassium dichromate solution, and the "iron hematoxylinophilic" granules were found to be composed of a nonsudanophilic protein-combining phospholipid detectable in paraffin sections due to the strong "iron hematoxylinophilic"nature of its protein portion fixed through the protein-precipitating action of the potassium dichromate acetic acid solution. The distributional peculiarities of these two sorts of granules in the glomerular, fasticular, and reticular zones of the adrenal cortex suggested that these granules would be closely related to adrenocortical endocrine function. In addition, the author could obtain extr act containing sudanophilic substances from the sudanophilic granules by means of a peracetic acid treatment followed by methylation, so that the author could messure a certain amount of ketosteroid content in the extract containing sudanophilic substances, using the colorimetric method based on the reducing reaction such as the technique of Nakao-Aizawa. These results as well as the above morphological findings, will support the conclusion that the iron hematoxylin-sudan III staining method will play a great part in the histochemical investigation of ketosteroids, as various histochemical test methods employed up to this time have been found valueless for the ketosteroids-detection.
The following results have been rendered reliable by means of the iron hematoxylinsudan III staining method, several histochemical dehydrogenase detection methods, and trypan blue as well as neutral red vital staining methods. On the 3rd day after enucleation of the adrenal gland, hyperemia in the capsule of the adrenal cortex and proliferation of fibroblast-like cells in the innermost side of the capsule were recognized, but no cortical cell regeneration could be found until the 5th day after enucleation when 3 β-ol dehydrogenase and G-6-P dehydrogenase activities become particularly noticeable in the regenerating cells. The new adrenocortical cells regene rate forming a fasticular structure divided off by the connective tissue cord towards the central part of the adrenal gland up until about the 30th day after enucleation. Moreover, the fact that among these cells at this stage of regeneration, cell groups with relatively few neutral red pigment assuming a glomerular structure, appear in the subcapsular region, may be regarded as constituting a new finding to support the opinion that the glomerular zone would be found during this stage. The various enzyme activities at this stage have been found to display considerable differences between cell groups in the subcapsular region corresponding to the glomerular zone and those assuming a fasticular structure, on the other hand, the difference in the form of arrangement between each of the two groups of cells corresponds approximately to that between these two zones in nontreated group, though to a less remarkable degree. These facts may be sufficient to suggest that the newly regenerated adrenocortical cells during the initial several days after enucleation, remain functionally and morphologically undifferentiated, but those from the 20th to the 30th day after enucleation, assume differentiation in their morphology. Moreover, judging from occurrence of cell groups undoubtedly not only morphologically but also histochemically corresponding to the glomerular zone, from the 40th to the 100th day after enucleation, it may be presumed that the functional and morphological differentiation of regenerated adrenocortical cells can proceed so far as to perfect a recreation of the enucleated adrenal cortex. The problem does remain, however, that even at this stage neith er the transitional nor the reticular zone has yet appeared, and that the medullary part of the adrenal gland remains wholly lacking. The newly re generating adrenocortical cells are considered capable of producing corticosteroids as early as the 5th day after enucleation, because of the remarkable activity of such enzymes as steroid 3 β-ol dehydrogenase and G-6-P dehydrogenase at that time. Accordingly, it is considered that the regenerated cells compensate the adrenoco rticosteroid secretion which is supposed to occur soon after enucleation of the adrenal gland.
The author studied macroscopically and histologically the process of reformation of the synovial membrane of the knee joints of rabbits in which synovectomy and capsulectomy were performed using as nearly as possible the same as Mori's technique. Specimen taken at various intervals following surgery was embedded in paraffin and sectioned and stained in a manner similar to that of Key (1925) and Wollcott) 1927). The technical difference of Mori's Synovectomy from that of Swett or Jones is th at in the former both joint capsule and synovial membrane is removed, whereas in the latter the synovial layer only is removed. In the author's experiments, therefore, contrary to the technique of Swett, the surgical intervention to the knee is more extensive through the capsular layer from the extra- or intra-muscular to sub-periosteal connective tissues. Our findings in this experiment fall into two groups: one is the reconfirmation of fac ts already established by the old pioneers which includes the following; 1. The reformation of the synovial membrane con be bu ilt in roughly 3 weeks. 2. The reparative process in the capsulectomized and synovectomized knee joint is essentially different from that of the surgical defect in skin or mucous membrane where the wound closure has to be accomplished by the extension of the e pithelial cells surrounding the original defect. In the synovectomized defect, h e lining cells are newly formed by the secondary differentiation of the proliferating co n nective tissue cells which are derived from the bottom of the defect. The second group is of new findings from this experiment which include the following; 1. The connective tissue participating in the reconstruction of the synovial lining is often derived form the peri-orintra-muscular tissue or sub-periosteal connective tissue located more deeply than the connective tissue in the outer layer ( Stratum fibrosum) of the joint capsule previously pointed out by Key as the single so u r se for synovial reformation. 2. The newly formed membrane is more fibrous in nature than the normal one. We suggest a new concept, however, that the change in the synovial membrane into a more fibrous and less vascular tissue can be regarded as evidence of convertion from an active to inactive (or from sensitive to resistant) tissue. The new synovial membrane therefore would be far less responsive to allergic inflammation. This agrees with our clinical experience that the long term follow up of synovectomized patients rarely if ever show subsequent involvement of the regenerated synovium in new inflammatory process.
The action of phenol on the central nervous system which is known to increase in the blood and the spinal fluid at the time of hepatic disturbances was studied on the rabbits. 1. The arousal reaction and muscular discharge due to a stimulation given to the thalamus and reticular formation were stimulated by administration of a small dose of phenol, while they were inhibited by a large dose of phenol. The same results were obtained as to the recruiting response and the spike and wave complex. 2. As the dose of phenol was increased, the frequency of the spontaneous brain wave tended to slow down, and also 4-5 c/s waves and triphasic wave became conspicuous. The spontaneous muscular discharge in the fore and hindlimbs tended to increase gradually as the dose of phenol was increased. These stimulating effects were observed even when the triphasic wave appeared. 3. The histologic al changes in the liver by phenol administration were those remniscent of an acute hepatic insufficiency. From these results, it will be concluded that phenol inhibits the consciousness by the inhibitory action on the ascending activating system and stimulates the motion by the action on the extrapyramidal system. Furthermore the comparativ e study was done between the administration of phenol and the ligation of the common bile duct. Phenol seems to be one of the factors which influences the symptoms occurring in the ligation of the common bile duct.
The case was 51-years-old male who had deformity of the spinal colum and marked ascites fluid. At au topsy, main pathological changes were found in the liver. In its cut surface, many yellowish nodules were seen, which were histologically identified as anemic infarct like necrosis of the hepatic cells coexisting with the picture of cirrhosis, although any morphological changes, such as thrombus, embolus or tumor, which may disturb the blood circulation, were found neither in hepatic artery, hepatic vein nor portal vein. It is generally believed that anemic necrosis of the liver are caused by the occlusion of hepatic artery. In this case, no morphological vidence of the occulsion in hepatic artery could be detected at autopsy. Therefore, the author assumed a functional occlusion of the hepatic artery in this case; that is, the deformity of the spinal colum, which narrowed the peritoneal cavity and displaced the hepatic artery, might have enhanced the compression and occlusion of the latter by ascites due to liver cirrhosis, followed by the anemic infarct like necrosis of hepatic cells.