Okayama University Medical School

The amygdalofugal fibers were studied III the cat with the silver method of NAUTA-GYGAX. 1. The amygdalofugal fibers are distributed by way of the stria terminalis, the longitudinal association bundle, the inferior thalamic peduncle, and the medial forebrain bundle. 2. The amygdalofugal fibers running through the longitudinal association bundle arise in the lateral principal, intermediate principal nuclei and the lateral and possibly intermediate parts of the periamygdaloid cortex, and terminate in the lateral preoptic nucleus, the bed nucleus of the anterior commissure, the olfactory tubercle, the nucleus of the diagonal band of Broca, the nucleus accumbens, the medial and posterior septal nuclei and the basal part of the head of the caudate nucleus. In addition, there are scattered fibers coursing along the longitudinal association bundle proper. These fibers may have a widespread origin from the amygdaloid complex. The longitudinal association bundle contributes no fibers to the medial forebrain bundle. 3. The fibers, originating from the lateral principal, intermediate principal and medial principal nuclei, join the medial forebrain bundle to distribute widely in the lateral hypothalamic nucleus. A few fibers are seen to reach the ventromedial hypothalamic nucleus, and are considered to arise in the medial principal nucleus. 4. By way of the inferior thalamic peduncle some fibers from the amygdaloid complex course dorsally into the medial part of the dorsomedial thalamic nucleus at its caudal levels. They may arise widely from the amygdaloid complex. A few of them extend farther dorsally to reach the lateral habenular nucleus and the parataenial nucleus. They probably originate from the lateral principal nucleus. 5. The fibers forming the stria terminalis originate from the medial principal nucleus, the medial nucleus, the periamygdaloid cortex and the cortical nucleus, and are distributed in the bed nucleus of the stria terminalis and the lateral preoptic nucleus (preoptic component), as well as the medial preoptic nucleus, the anterior hypothalamic nucleus and the ventromedial hypothalamic nucleus (supracommissural component). The cortical nucleus, particularly its caudal part, and possibly the medial part of the periamygdaloid cortex are regarded as the main sources of the stria terminalis fibers ending in the hypothalamic region. The intermediate principal and lateral principal nuclei do not appear to contribute fibers to the stria terminalis. 6. The ventromedial hypothalamic nucleus receives amygdalofugal fibers both from the medial principal nucleus by way of the medial forebrain bundle, and from the cortical nucleus via the stria terminalis. 7. In addition to intrinsic internuclear fibers within the amygdaloid complex, some of the fibers from the complex are distributed to the ventralmost part of the putamen, the medial part of the claustrum, the periamygdaloid cortex, the prepiriform area and the anterior amygdaloid area, but do not reach the hippocampus.

This study was carried out to determine whether an extracorporeal membrane oxygenation (ECMO) support could be sufficiently conducted by the right ventricle alone from the viewpoint of the hemodynamics and blood gas state. Six infant dogs underwent a bypass between the left pulmonary artery and left atrium with an in-line oxygenator after a left pneumonectomy. Partial ECMO support was conducted simply by opening the circuit, and total ECMO support was conducted by ligating the right pulmonary artery. After the establishment of partial ECMO, approximately one-third of the right ventricular output was passively shunted through the bypass circuit, and the cardiac index and central venous pressure did not change. The mean pulmonary arterial pressures increased significantly. After a complete ligation of the right pulmonary artery, all 6 dogs survived for 12 h, but the cardiac output and blood pressure decreased significantly. The blood gas state was sufficiently maintained throughout the experiment. The results suggest the possibility of using the pumpless ECMO support. However, the flow resistance of the membrane oxygenator proved to still be too high for use in a total pumpless ECMO. Further studies on long-term ECMO and the development of a membrane oxygenator with a considerably low flow-resistance are needed.

With the purpose to elucidate morphologically the site where fat synthesis takes place in the cell, electron-microscopic observation has been conducted on the interscapular brown fat tissue of mice at various periods of carbohydrate introduction after starvation. By starving mice, the depot lipids in the brown fat have been discharged almost completely, and the carbohydrate introduction has caused the biosynthesis of lipids from carbohydrtates in the same tissue. Observations on the tissues proved that the lipogenesis in the brown fat tissue cells takes place in the ground substance keeping the intimate correlation with the endoplasmic reticulum but not in the mitochondria.

An electron microscopic study on the fine structural differences of motor endplates among the red, white and intermediate muscle fibers of the rat intercostal muscles was made and the following results were obtained. 1. In the motor endplate of the red fiber, the junctional folds were poorly developed and their number was small. 2. In the motor endplate of the white fiber, the junctional folds were well developed and their number was far more numerous than those in the red fiber. 3. The fine structure of the motor endplate of the intermediate fiber was of an intermediate character between the red and white fiber.

An electon microscopic study on the structural differences among the red, white and intermediate muscle fibers of mice was made and the following results were obtained. 1. The red fiber contained very numerous mitochondria, the white fiber a few and the intermediate fiber a moderate number. The distribution of mitochondria was different in each type of muscle fiber. The cristae of mitochondria of the red fiber was quite well developed, that of the white fiber poorly and that of the intermedtate fiber moderately. 2. Sarcoplasmic reticulum of the white fiber was considerably well developed but that of the red and intermediate fibers poorly developed. 3. Glycogen particles were abundant in the white fiber, less in the intermediate fiber and least in the red fiber.

Electron microscope study on the rat liver cells of carbon tetrachloride poisoning has been reported. Observations have been made on the osmic fixed tissue sections obtained from the liver at early stages of poisoning, 5 to 22 hours after carbon tetrachloride oral administration, 0.25ml. per 100g. body weight. Special attention is paid on the appearance of electron dense area, opaque area, in cytoplasm, which is compased of fibrous components, probably originated from endoplasmic reticulum. This will be an important sign of cell degeneration. Toluidine blue, PAS and methyl green-pyronin stainings of the thicker sections from the same samples as used for electron microscopy revealed that the opaque area is stained by toluidine blue and pyronin but not by PAS. The opaque areas appear already five hours after the carbon tetrachloride administration and show some continuity with elongated filaments of endoplasmic reticulum. At an advanced stage of paisoning the opaque area increases in its number and size, but some of them are shrunk as a mass, being separated from the surrounding cytoplasm with scanty area. Often they form denser masses in the center and loak like the lipid deposition. The picture suggests formation of lipid droplets in the case of fatty degeneration of the liver cell.

1. The studies of structure and function of the plasma membranes of cancer cells is extremely important for the elucidation of specificity of phenotypes of cancer cells. In order to bring this subject to light, plasma membranes, mitochondria, microsomes and nuclei have been isolated from the AH 130 ascites carcinoma cells and rat liver cells. The electron cytochemical observations and biochemical assays of M g²+-Na+-K+-ATPase, ADPase, AMPase, and β-glycerophosphatase activities have been carried out before and after the fixation with glutaraldehyde. 2. M g²+-ATPase and Mg²+-N a +-K +-ATPase are present in the isolated plasma membranes, mitochondria and microsomes in both AH 130 cells and rat liver cells. ADPase and AMPase of the mitochondria and microsomes show far lower activities than those of the corresponding enzymes found in rat liver plasma membrane. ADPase and AMPase of AH 130 cell fraction exhibit activity much lower or zero. Generally, enzymatic activity of the AH 130 cell fraction is much lower than that of rat liver cell fraction. 3. Mg²+-Na+-K+-ATPase is completely abolished by 5% glutaraldehyde fixation while it shows less effect on Mg²+-ATPase in the plasma membrane. ADPase and AMPase activities of the mitochondria and microsomes are completely inhibited by glutaraldehyde fixation. AMPase of the plasma membrane of rat liver is completely abolished while ADPase activity is not affected in any way. 4. Only Mg²+-ATPase can be demonstrated electron cytochemically. Cytochemical reaction products of Mg²+-ATPase are located at the outer layer of the plasma membrane of the AH 130 cells and rat liver tissue. In the isolated membrane fractions it is located at the inner layer. 5. ρ-Chloromercuribenzoate has only a slight effect on Mg²+-ATPase and Mg²+-Na+-K+-ATPase activities of the rat liver membrane, while it inhibits these enzyme activities in the AH 130 cell membrane. NaF (1 mM) and NaN3 (1 mM) inactivate ADPase of the rat liver plasma memo brane. 6. In these experimental conditions, nonenzymatic hydrolysis of ATP by lead ions is not recognized. 7. It seems most reasonable to conclude that cytochemical electron microscopic demonstration of Mg²+-ATPase after fixation with glutaraldehyde may serve as the absolute marker for the plasma membrane of ascites hepatoma and liver cells.

An autopsy case of spinal arteriovenous malformation (AVM) was reported. The patient was a 75-year-old male and his initial neurologic symptoms were paraplegia, paresthesia below the umbilical level and urination difficulty. Subsequently night delirium and parkinsonism also appeared. The clinical and pathological findings in this case are identical with those in the spinal AVM except for Parkinson's disease. In addition, the lateral funiculus of the spinal cord in the middle thoracic segment showed pallor: Under light microscopy, the funiculus was spongiform, with a thinner wall of the myelin sheath, enlargement of the axon and the perivascular infiltration of phagocytes without plasma exudation. The changes in the lateral funiculus seemed to indicate early congestive changes.

An autopsy case of hypertrophic cardiomyopathy showing clinical features of dilated cardiomyopathy was reported. The patient was a 60-year-old female complaining of chest discomfort from the age of 40. At autopsy, both ventricles were dilated. Microscopically myocardial loss, fibrosis and disarray of hypertrophic myocardial fibers were observed. The areas showing myocardial disarray were distributed close to the scar-like fibrotic areas. Coronary arteries and intramyocardial arterioles showed minimal stenotic changes.

The elimination of fibroblast-like cells from primary cultures of fetal human livers was studied. A fibroblast-like cell line (HuF), which was obtained by subculturing fetal human liver cells 4 or more times, was briefly treated with hydrocortisone (HC) or putrescine (PUT). The growth of HuF cells was inhibited by HC at a concentration of 10(-2) M and by PUT at a concentration higher than 10(-3) M. Long-term treatment of HuF cells with 10(-3) M HC inhibited the growth of the cells. Primary cultures of fetal human livers were made in medium containing HC or PUT, and morphological and functional examinations were made. The cultures were predominantly composed of epithelial-like cells, with few fibroblast-like cells, when the HC concentration was 10(-5)M to 10(-3) M. A high amount of albumin was secreted at these concentrations of HC. On the other hand, at 10(-3) M PUT, many epithelial-like cells were seen, but albumin was undetectable. The present results indicate that albumin-producing epithelial-like cells can be selectively maintained in medium containing HC, in primary cultures of fetal human livers.

A mixture with essential and nonessential amino acids high in branched chain amino acids and low in aromatic amino acids (Fischer solution), and another synthetic mixture of branched chain amino acids containing 3 amino acids associated with the urea cycle (Hep-OU) were infused to control subjects and patients with severe hepatic disease. Alterations in serum aminograms, blood ammonia levels and electroencephalograms following the infusion were studied and compared with those obtained by a commercially available amino acid mixture. Short-term or continuous infusion of a commercially available amino acid solution to cirrhotic patients caused an increase in methionine, phenylalanine and tyrosine and a decrease in branched chain amino acids. These post-infusion results were similar to the patterns seen in hepatic encephalopathy. In cirrhotic patients, infusion of Fischer solution which contains small quantities of methionine and phenylalanine produced an increase in the concentrations of these 2 amino acids, probably because of impaired utilization by the injured liver. No marked alterations in serum aminograms, however, were observed in cirrhotic patients either immediately after, or 3 h after, the end of the Hep-OU infusion. Reduction of methionine, tyrosine and phenylalanine levels and elevation of the molar ratio of (valine+leucine+isoleucine) / (phenylalanine+tyrosine) were significant. The infusion of Hep-OU to patients with liver cirrhosis or subacute hepatitis resulted in clinical and neurological improvements and the restoration of the molar ratio of branched chain amino acids/aromatic amino acids.

1. An apparatus for the simultaneous measurements of volume change, fluorescence intensity of pyridine nucleotides and oxygen consumption of mitochondria has been constructed. 2. Oxygen consumption is measured by the rotating platinum electrode with a modification of Hagihara's system, attached in a cuvette of the apparatus. 3. Volume changes of mitochondria (swelling-shrinkage) are measured by the 90° light-scattering at 650 mμ. 4. Relative fluorescence intensity of pyridine nucleotides is measured by the fluorometer: for the excitation, a bright light at 365 mμ. line of mercury lamp is isolated through the filter and exposed to the mitochondria suspended in a cuvette of the apparatus, and fluorescent emission is analyzed by a grating mirror monochromator. 5. The scattered light at 650 mμ. is not affected by the excitation light and the fluorescent emission, and fluorescence intensity is not affected by the scattered light at 650 mμ. 6. The simultaneous measurements of the oxidation-reduction of pyridine nucleotides, the respiration states and the changes in the intensity of 90° lightscattering of mitochondria are given as an example of the performance of the present apparatus.

A reproducible animal model of fulminant hepatic failure was developed by intraperitoneal administration of D-galactosamine hydrochloride to Sprague-Dawley rats. Biochemical and morphological hepatic injury and brain edema resembled human fulminant hepatic failure. This model would facilitate further studies of the pathogenesis of brain dysfunction and evaluation of treatment in fulminant hepatic failure.

In order to determine the steps with which the reaction of neotetrazolium chloride reduction conjugates in the terminal electron transport system, an analytical study on the neotetrazolium reduction by tissue homogenates was carried out using various substrates such as sodium succinate, p-phenylenediamine, sodium malate, sodium α-glutamate and DPN, and inhibitors such as sodium malonate, potassium cyanide and antimycin A, as the results the following conclusions were drawn. 1. The reaction of neotetrazolium reduction by tissue homogenate using sodium succinate as substrate is mainly the succinoxidase system reaction; and the reaction takes place conjugating about 50 per cent in the step of the succinic dehydrogenase system (succinic dehydrogease, cytochrome b and cytochrome C1), of these about 15 per cent conjugates in the step prior to the antimycin A sensitive step and 35 per cent in the step itself; and about 50 per cent in the step of cytochrome c oxidase. 2. In the case using p-phenylenediamine as substrate the reaction of neotetrazolium reduction is the reaction due to the activity of cytochrome c-cytochrome oxidase system; and when p-phenylenediamine is used with the sufficient amount of cytochrome c, the reaction appears to be dependent on cytochrome c oxidase activity. Neotetrazolium reduction in all these reactions takes place conjugating in the step of cytochrome c oxidase. 3. In the case where DPN and substrates taking DPN as a coenzyme are used, the reaction of neotetrazolium reduction is mainly the reaction conjugating at the step below antimycin A sensitive step in the DPNHcytochrome c reductase system (flavoprotein, cytochrome b and cytochrome c;), probably with the flavoprotein of DPNH-dehydrogenase. 4. Endogenous dehydrogenase reactions are the sum total reactions conjugating at the steps prior to the antimycin A sensitive step in the terminal electron transport system and with other various reduction systems which are not inhibited by antimycin A.

To determine whether a relationship exists between DNA ploidy and the prognosis of hepatocellular carcinoma (HCC), flow cytometric DNA analysis was performed in paraffin-embedded specimens obtained from 44 patients with HCC who underwent hepatectomy. There were 26 diploid (59%) and 18 aneuploid (41%) tumors. No correlation was shown between DNA ploidy pattern and patient age, sex, liver cirrhosis, hepatitis B virus antigen and serum alpha-fetoprotein level. The ploidy pattern had no significant correlation with the presence of vascular invasion or intrahepatic metastasis. Only Edmondson's grade was well correlated with the ploidy pattern. We noted a significant correlation between survival rates and the presence of vascular invasion or intrahepatic metastasis (p < 0.05). In contrast, no significant correlation was found between DNA ploidy pattern and the prognosis of HCC. The results of this study indicate that DNA ploidy pattern may not be a useful indicator for the prognosis of HCCs after hepatic resection, unlike the results of gastric and colon cancers.

A 31-year-old female with chronic myelogenous leukemia, who developed myeloblastic involvement of the central nervous system during acute myeloblastic transformation of the disease, was treated with methotrexate intrathecally. The therapy produced prompt clinical response and complete reversal of abnormal cerebrospinal fluid findings. However, the patient expired 10 months following the acute blastic crisis.