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To investigate the involvement of actin filaments in concanavalin A (Con A)-induced cap formation and cytochalasin B (CB)-induced zeiotic knob migration, the distribution of F-actin was studied in Con A-treated and CB-treated Ehrlich ascites tumor cells (EATC) by fluorescence microscopy using heavy meromyosin conjugated with a fluorescent dye, N-(7-dimethylamino-4-methylcoumarinyl) maleimide, (DACM-HMM). In non-treated cells, the diffuse fluorescence of DACM-HMM was observed in the cytoplasm, particularly intensely under the plasma membrane and around the nucleus. In Con A- and CB-treated cells, the fluorescence was seen at Con A-induced-capped and CB-induced-knob-accumulated regions. This fluorescence was more intense in CB-treated cells. To study the actin filaments in these fluorescent regions more clearly, the soluble components of the cells were eliminated by treatment with Triton X-100 or saponin solution containing a low concentration of glutaraldehyde, and the detergent-treated and saponin-treated cells were observed under a transmission electron microscope. Concentrated actin filaments were observed directly beneath the Con A-induced capping area and CB-induced zeiotic knob-accumulation area. The area of concentrated actin filaments appeared to correspond to the electron dense area observed in the identical region in the cells fixed without detergent treatment. More actin filaments were observed in CB-treated cells than in Con A-treated ones.

The effects of ethanol on rat Kupffer cells were studied functionally and morphologically. Eight g ethanol per kg body weight per day was intragastrically administered to rats for 7 days. An isocaloric glucose solution was administered to control rats. The phagocytic activity of the reticuloendothelial system was measured by the carbon clearance method (57 mg carbon particles per kg body weight) on the 7th day. Kupffer cells having phagocytized carbon particles were counted under the light microscope. Kupffer cells were also observed by scanning electron microscopy. Both the carbon clearance and Kupffer cell number were lower in ethanol-administered rats (32 +/- 8 X 10(-4) mg/ml; 0.6 +/- 0.3/0.01 mm2 liver lobule) as compared to control rats (63 +/- 15; 3.1 +/- 1.0). Microvilli and filopodia of Kupffer cells were fewer in ethanol-administered rats than in control rats. Carbon clearance correlated with Kupffer cell number per 0.01 mm2 liver lobule and liver weight. These results suggest that the decrease in carbon clearance induced by ethanol is due mainly to the decrease in Kupffer cell number and partly to the decrease in Kupffer cell activity as demonstrated by the disappearance of microvilli and filopodia.

Lymphocyte activation by streptolysin O (SLO) and factors in the plasma which inhibit the response to SLO were examined in 19 patients with mucocutaneous lymphnode syndrome (MCLS), 54 age-matched (6 months-6 years) normal children, 41 normal children older than 6 years and 10 normal adults. In normal children younger than 6 years, the response to SLO was weak and in many cases no response was seen. On the other hand, in the patients with MCLS, the response of lymphocytes to SLO was high and comparable to the response in adults and children older than 6 years. The DNA synthesis of lymphocytes stimulated by SLO was inhibited almost completely by autologous or allogeneic plasma of many of the normal children and adults. The plasma of patients with MCLS did not inhibit, but rather enhanced the response to SLO. These results suggest that the increased response of lymphocytes to SLO and the lack of plasma inhibitory factors in patients with MCLS may be due to the immune response to the pathogen of MCLS, as yet undiscovered.

The replicative responses of suckling and adult rat hepatocytes in primary culture to growth-stimulating factors were compared. By addition of L-proline alone, the [3H]-thymidine labeling of suckling rat hepatocytes was dramatically enhanced, but that of adult ones was not. Epidermal growth factor (EGF), insulin, triiodothyronine (T3) and glucagon also enhanced the labeling of suckling rat hepatocytes regardless of the presence or the absence of L-proline. On the other hand, in the absence of L-proline, only EGF enhanced the labeling of adult rat hepatocytes, and, in the presence of L-proline, insulin as well as EGF enhanced the labeling. In the presence of growth factors and L-proline, the number of suckling rat hepatocytes increased up to about 143%, whereas that of adult rat hepatocytes hardly increased. Thus, a remarkable difference in replicative responses to growth factors and L-proline was observed between suckling and adult rat hepatocytes in primary culture.

Primary cultures of surgically obtained noncancerous portions of human liver tissues were made. Liver tissues were poorly dissociated with collagenase, but well dissociated with dispase. The yield and viability of cells were improved somewhat when dissociated with collagenase followed by dispase. The mean cell yield was 1.1 X 10(6) cells/g liver. The epithelial-like morphology of the dissociated liver cells was maintained for about one week, but thereafter degenerative alteration of cells was observed. In liver explant culture, an active outgrowth of cells was observed for more than one month. Albumin production in culture fluids from dissociated livers was detectable for about 2 weeks, but later became undetectable, while that from explant culture was detectable for at least one month. These data demonstrate that adult human hepatocytes can be isolated from noncancerous portions of livers with relatively high yield, and that albumin production of the dissociated cells is detectable for several days.

To study chromatin structure at the sites of DNA replicated in permeable cells, deoxyribonuclease I (DNase I) sensitivity of newly replicated DNA in permeable mouse sarcoma cells was compared with that of newly replicated DNA in intact cells. About 35% of the DNA replicated in permeable cells was hypersensitive to DNase I, and the remaining DNA showed the same DNase I sensitivity as that of parental chromatin DNA. The sensitivity of DNA replicated in permeable cells was higher than that of DNA newly replicated in intact cells, and was close to that of DNA replicated in the presence of cycloheximide. The sensitivity of DNA pulse-labeled with [3H]deoxythymidine triphosphate by replication in permeable cells was reduced significantly by chasing with cold deoxythymidine triphosphate. The present results suggest that chromatin structure at the sites of DNA replicated in permeable cells is similar to that at the sites of DNA replicated in living cells in the absence of protein synthesis, and that some structural change (possibly toward the maturation) of newly replicated chromatin occurs after the DNA replication in permeable cells.

Development of the ear, especially the middle ear, was studied histologically in ddN and CF mice. Primordia of the 3 ossicles and the otic capsule appeared on day 12 of pregnancy. The stapedial primordium was observed as a mass of mesenchymal cells lateral to the primordium of the otic capsule, attaching to the medial part of the facial nerve. On day 13, the stapedial primordium continued to develop with the Reichert's cartilage. On day 14, the malleus and incus were differentiated. On day 15, the 3 ossicles were mostly completed in shape, and the stapedial footplate had a bilaminar structure at this stage. This structure appeared to correspond to the lamina stapedialis in the developing human stapes.

This study was designed to determine the in vitro release of tegafur from a suppository and the in vivo bioavailability of tegafur in rats. Two different suppository preparations (product A-1 and product A-2) containing 750 mg of tegafur were tested for in vitro release of tegafur by the Muranishi Method (membrane diffusion method) and the partially modified paddle method (permeability through dialysis tubing). When determined by either method, the amount of tegafur released from product A-2 during the whole experimental period was significantly greater than that released from product A-1. When tested by the Muranishi method, however, the difference in the amount released during the first 10-min period was not significant. A greater bioavailability of tegafur after rectal administration was obtained by product A-2 more than product A-1. A significant correlation was observed between the in vitro release and the in vivo bioavailability. The present results indicate that there are considerable differences in physiochemical characteristics between product A-1 and product A-2.

Liver biopsy specimens obtained from a 31-year-old female with delta-positive hepatitis were studied by routine electron microscopy. In several nuclei of hepatocytes, there were filamentous or microtubular structures 15 to 20 nm in diameter, in the vicinity of which, round particles, probably cross sections of tubular ones, were seen. In these nuclei, irregular granules approximately 20 to 30 nm in diameter were also found in clusters. However, cores of Dane particles were not found in such hepatocytes. These intranuclear microtubular structures may be associated with delta agent.

Maximal expiratory volume-time and flow-volume (MEVT and MEFV) curves were drawn for young male nonsmoking healthy adults and for young male nonsmoking asthmatic patients. Eleven parameters, two MEVT (%FVC and FEV1.0%), six MEFV (PFR, V75, V50, V25, V10 and V50/V25), and three MTC parameters (MTC75-50, MTC50-25 and MTC25-RV) were used for the multivariate analysis. The multivariate analysis in this study consisted of correlation coefficient matrix computation, the test for mean values in the multivariates, and the linear discriminant analysis using the all possible selection procedure (APSP). Correlation coefficients among flow rate parameters and flow rate related parameters in high lung volumes were different between the two groups. In the eleven-parameter discriminant analysis by APSP using single parameters, PFR, V75 (flow rate at 75% of forced vital capacity), and FEV1.0% were considered to be the effective parameters. In the seven-parameter discriminant analysis using the parameter groups, the group of all parameters and the %FVC and flow rate-related parameter group were considered to be the effective numerical alternatives to MEFV curves discriminating between healthy adults and asthmatic patients.

With the purpose of revealing the biological effects of the X-ray irradiation the authors extracted phospholipids from the liver of irradiated animals and proved that this substance has the action to inhibit the growth of the bone marrow cells, the motility of pseudo-eosinophilis and the erythropoiesis in tissue culture, suggesting that the injury will mainly be induced by the toxic substances produced by irradiation.

1. The DNA contents in mature lymphocytes of the mouse, rat and man are kept almost constant. 2. The variety in the DNA contents in tumor cells is attributed to the rapid DNA synthesis taking place at the interphase, though the degenerating cells and the cells in abnormal mitosis can not be discarded as the source of the variety in DNA content. 3. The RNA content in AH-130 (ascites hepatoma) is less than that in normal liver cells.

Leukemic cells were cytologically studied in the human bone marrow culture by the utilization of vital staining of Janus green B and neutral red. The minute cellular morphology of various types of leukemia was studied with special reference to their alterations in the course of the culture. The cytologic deviation of leukemic cells from the corresponding normal blood cells was clarified on monocytic leukemia, chronic myelogenous leukemia with the blastic crisis, chronic myelogenous leukemia, and acute and chronic lymphocytic leukemias.

Vital observation on the cellular morphology of the normal human blood cells was conducted by means of bone marrow culture successfully in conjunction with vital staining with Janus green B and neutral red. A special attention was paid for the alterations of the cellular structures in the course of the culture. The findings are summarized as follows : 1) Intracellular particles with affinity to Janus green B or neutral red were classified into minute granules, granules, vacuoles, and mitochondria. Morphologic features of each type of the particles were studied in detail. 2) Two types of granules are present in neutrophilic and eosinophilic blood cells, whereas one type of granules is present in basophilic blood cells. Eosinophilic and basophilic granules show characteristic pole formation in them at the terminal stage of the staining. 3) The rosette formation in the mature monocyte and the aggregations of neutral red vacuoles in the mature neutrophil and the mature lymphocyte were characterized. 4) The cluster of neutral red vacuoles is characteristic of the erythroblast. 5) The mitochondria of the mature neutrophil and the mature monocyte participate in producing neutral red vacuoles.

1) The influence of thyroid secretion upon the induction of leukemia in Dba/2 male mice by methylcholanthrene was investigated. Radiothyroidectomy significantly reduced the incidence of leukemia in these mice. This reduction in incidence did not occur if radiothyroidectomy was performed after the administration of the carcinogen. 2) Data indicated that hypothyroidism following radiothyroidectomy interfered with the initiation rather than the promotion of methylcholanthrene- induced-Ieukemogenesis. 3) No correlation between incidence of leukemia and body weights in the mice was noted.

1. The unsaturated fatty acid fraction (OX) from the liver of irradiated rabbits contains substance which has the same effects as X-ray irradiation on the testicular cells. 2. This substance introduced intravenously causes the degeneration of the germinal cells with the formation of giant cells or multi-nucleated cells and the mitotic abnormalities. 3. The DNA content of the cell also shows the changes exactly identical with that seen after X-ray irradiation. 4. From these results we conclude that the X-ray injury will be mainly due to the production of some toxic substance which is found in the unsaturated fatty acid fraction and severely affects the cells in mitosis and DNA metabolism.

The unsaturated fatty acid fraction extracted from the liver of rabbit irradiated with X-rays exerts a strong cytotoxic effect on human coelothelioma cells and Yoshida sarcoma cells both in vitro and in vivo. The cell damage seems to initiate at the nucleus, finally leading to the complete cytolysis. The inhibiting effect of this substance on the mitosis of Yoshida sarcoma cells can be observed, especially marked from prophase up to metaphase giving almost the same results obtained after X-ray irradiation. From these results and the observations reported by several authors on the cell damage by X-ray irradiation, weshould call special attention to the fact that the essential mechanism of X-ray irradiation can be attributed to the cell toxin produced after the irradiation.

Within the range of our investigations the most important biochemical characteristics in the brain of idiopathic epileptic patients seem to be defect in the production of and the attendant decrease in free amino acids of the brain. On account of these phenomena there seem to occur the acceleration of the ChE activity and a poor utilization of glucose. Of the free amino acids in the brain the combined amount of glutamic acid, glutamine and γ-aminobutyric acid (GABA) will occupy the major portion of the total free amino acids found in brain, and thus diminution in the contents of glutamic acid and GABA in the brain of idopathic epileptic patients has quite an important meaning. At the present stage it is not yet possible to give any definitive answer to the question why such decrease occurs but it is believed that the most urgent problem facing us today is the amino acid metabolism that is associated with glutamic acid and the comparative studies of the amino acid metabolism in the epileptic brain to that in the non-epileptic brain are required. The fact that γ-amino-β-hydroxybutyric acid (GABOB), the substance that suppresses the central excitation, is decreased seems to indicate biochemically the existence of a defect in the processes of excitation in the brain of idiopathic epleptic patients.

1. The results of 245 pleural biopsies perfomed in 108 patients including 219 pleural needle biopsies and 26 pleural open biopsies were reported. The method of pleural biopsy seems to be superior to any other currently available diagnostic procedures for the etiological diagnosis of pleurisy. 2. When the pleural needle biopsy is compared with the pleural open biopsy, the former method has definite advantages over the open biopsy. The pleural needle biopsy is simple, repeatable and has almost no complication. The method of pleural needle biopsy is the initial method of choice as Donohoe correctly stated and should be employed in every cases of the pleurisy to confirm the etiological diagnosis. The open biopsy should be reserved only for those cases in whom the needle biopsy had not proved satisfactory. 3. Utilizing the method of needle biopsy, the pathological diagnosis was made in 86 per cent of our cases at the initial biopsy. By repeated needle biopsies, the results have improved to 91-92 per cent. 4. Most of the failures of the pleural needle biopsy were noted at the early stage of the study due to the unfamiliarity of the biopsy technique and later due to the incooperation of the patients. 5. The presence of the free pleural fluid serves as a convenient guide for the performance of the needle biopsy but successful needle biopsy was easily done without presence of pleural fluid when there is adequate pleural thickening. 6. 63-75 per cent of our diagnosed cases were proved to have granulomatous pleuritis, 13-31 per cent non-specific pleuritis and 5.4-5.8 per cent eosinophilic pleuritis due to paragonomiasis. The distribution of this pathological diagnosis seems to reflect quite well the actual picture of incidences of pleurisy of various different etiology in young adults in Korea. 7. The relationship of the success in obtaining adequate tissue by needle biopsy and interval between onset of symptom and biopsy was discussed. It was found that the interval has no significant effect on the production of adequate tissue by needle biopsy if the time elapsed is 4 weeks or more from the onset of symptom. 8. The significance of the pathological findings of ranulomatous pleuritis at one biopsy and non-specific pleuritis at another biopsy in the same patient was discussed. It is concluded that the single finding of nonspecific pleuritis at one needle biopsy cannot rule out the presence of granulomatous pleuritis and it is recommended that pleural biopsy be repeated whenever necessary. 9. The diagnostic significance of the chemical analysis of the pleural fluid was discussed in correlation with the results of the pleural needle biopsies. It is concluded that the number of examinations are not quite sufficient to draw any definite conclusion at the present stage of our study. 10. The finding of sanguinous pleural fluid in the patient of granulomatous pleuritis is quite high (72.7 %) and it was found that the sanguinous pleural fluid was most frequently found in the patients with granulomatous pleuritis in non-cancerous age. 11. Two groups of pleurisy patients with or without parenchymal lung lesion on chest X-ray were discussed in correlation with the results of the needle biopsy. It was found that the incidence of the pathological evidence of granulomatous inflammation on the biopsy specimens in these two groups is almost the same regardless of the presence of the demonstrable parenchymal lung lesion. 12. Histopathological finding of granulomatous pleuritis was discussed in conjunction with the significance of two types of tubercles, the soft tubercles and hard tubercles. In all specimens diagnosed as granulomatous pleuritis granulomas were demonstrated ranging from large, conglomerate tubercles with central caseation or giant cells to small granulomas without central caseation or Langhans' giant cells. 13. Histopathological significance of the finding of non-specific pleuritis on the biopsy specimens was discussed and the existence of a specific entity of "non-specific pleuritis" which is equivalent to the non-specific inflammation of the pericardium. 14. Cases of pleurisy due to paragonomiasis were discussed and the need of specific attention for search of new cases was emphasized.