A fibrob'ast-inhibiting agent, chloropuine, was employed to be treated against experimental animal tumors, Brown-Pearce cancer and Yoshida sarcoma. The results are as follows 1) A higher survival rate of the rabbits bearing Brown-Pearce cancer and inhibition of the growth of the tumor were observed by daily administration of chloroquine diphosphate of 10 mg/kg. However no significant results were obtained in the animals with far advanced cancerous involvement. 2) No significant life prolongation was obseved in the rats bearing Yoshida sarcoma by chlcroquine administration of daily dosis of 25 mg/kg, but this ttreatment appeared to be inhibited accumulation of the ascites and increase of the ascites tumor cells. 3) As shown above, a fibroblast-inhibiting agent, chloroquine, used in the treatment of the animal tumors, led to reasonably good result.
Because catalase activity is thought an important indication in chloroquine efficiency against malignant tumors, changes of the catalase activity of the liver in the animals bearing malignant tumors was observed by chloroquine administration. 1) No significant changes were noted in the catalase activity of the liver of the normal healthy mice by administration of chloroqujne diphosphate of daily dosis of 25 mg/kg. 2) It was clearly demonstrated that catalase activyit of liver tissue of the normal mice was scarcely influenced to direct addition administration of 200γ/ml/hr of chloroquine diphosbhate. 3) In the mice beariog Bashford cancer given daily dosis of 25mg/kg of chloroquine diphosphate, lowering of liver catalase activity was observed to be considerably inhibitory in the first week and tended to show more remarkable with passage of the dates. 4) No inhibition of the decreasing liver catalase activity was observed by one week administration of chloroquine diphosphate in the mice bearing Ehrlich carcinoma. 5) Decreasing of the liver catalase activity was suppressed by one week administration of chloroquine in the rats bearing Yoshida sarcoma. 6) No inhibition of the decreasing Iiver ceatalere activity was observed by 2 weeks administration of 10 mg/kg of chloroqujne diphosphate in the rabbits bearing Brown-Pearce cancer which dimonstrated far advenced canceraus involvemeut and ascites accumulation.
In experimal animal bearing tumors such as Bashford cancer and Yosida sarcoma, abnormality of iron metablolism was investigated especially in their organ iron. The rerults are as follows 1) In normal mice, changes of the non-hemin iron of the organs were scarcely noted by chloroquine administration and it may be thought that the agent does not have any signigicant influence to the iron metabolism in the normal mice. 2) It tended to show that inhibition of the storage of the non-hemin iron in the tumor tissues or organs of Bashford cancer and Brown-Pearce cancer which were shown the inhibition of the tumor growth. Also a tendency of the decresing iron content of the ascties in mice bearing Ehrhich cancer which was failed to inhibit the growth of tumor and prolongaton of life. 3) It is presumed from these data that chloroquine shows anti-tumor activity in the iron metabolism of the animals bearing tumors.
Nearly all the investigations on abnormalities of the gastric and duodenal motility in acute hepatitis have depended on fluoroscopic findings. However, the fluoroscopic method has disadvantages in that it is not possible to observe the motility of the stomach and duodenum for any length of time nor can it afford quantitative estimation of the motility, resulting in the subjective decision of the examiner, In view of this, the author took “balloon kymographic recordings” of antral gastric and duodenal motility in 6 normal persons, 10 cases of acute hepatitis, 12 of chronic hepatitis, and 6 of recurrent chronic hepatitis. On the basis of these recordings, the general pattern of motility, description of waves and analysis of the records are given in the present communication. 1. In the normal control group the total activity (percent of time present) of antral gastric motility proved to be 38.2±2.6%. Of it 23.5±2.5% was consisted of the twenty-second rhythm described by Carlson, while the rest was composed of nonrhythmic waves. The total activity of duodenal motility was 60.6±2.7%, and the waves seen in the records were classified into two types, designated Types I and III. The percent of time taken by nonrhythmic Type I wave was 31.3±2.3% and that by Type III wave, 27.2±2.3%. 2. In the icteric stage of acute hepatitis, the total activity was decreased both in antrum pylori and duodenum and mean amplitude (cm water) tended to rise, while the standard deviation of mean amplitude was large. In the motility pattern of duodenum there was recognized a rising tendency in the basal pressure of Type III wave. At the recovery stage where jaundice had disappeared, both the general pattern of motility and the total activity in antrum pylori and duodenum returned normal. 3. In chronic hepatitis there could be observed no significant dlifference from normal control group. In recurrent chronic hepatitis there could be seen a decrease in the total activity just as in the case of acute hepatitis at icteric stage. However, there was no significant difference in mean amplitude. 4. In two of the ten cases with acute hepatitis at icteric stage and in two of the six cases with recurrent chronic hepatitis, the balloon did not pass through pylorus, and this is judged to be due to motility abnormality of pylorus. 5. Mean duration of the waves as observable in motility patterns of antrum pylori and duodenum proved to be the same both in acute and chronic hepatitis as well as in normal control. 6. The reduction of motility observed in stomach and duodenum may be pointed out as one of the causes for discomfort of the upper abdomen in acute hepatitis at icteric stage and in chronic hepatitis at recurent stage.
For the purpose to study histopathological conditions of the duodenum in acute hepatitis, biopsy examinations of the mucosa at the third part of duodenum were done under fluoroscopy by the intestinal biopsy capsule after Crosby. The subjects of the present study were 5 cases of normal controls and 12 patients with acute hepatitis admitted to our clinics, and on the latter group similar examinations were given at two stages; one at icteric stage and the other at convalescent stage. Biopsy specimens were stained with hematoxylin-eosin, Azan stain, Pap's method of a modified form of Bielschowsky's silver impregnation, Van Gieson's stain, and PAS-Schiff reaction. Each group of these specimens were prepared into 100 serial sections of 15μ in thickness, and theses were examined according to the following items: The length and width of villi, the arrangement and morphology of epithelial cells; the abnormality in the array of epithelial cell nuclei; the number of lymphocytes within vacuoles of epithelium; the number of goblet cells among epithelial cells; breakages of basement membrane: the lymphocytes in tunica propria; infiltration of plasmocytes, the thickness of mucosa, mitotic index in lacuna; and arrangement and morphology of epithelial cells in crypt; the number of lymphocytes in epithelial cells, the number of goblet cells among epithelial cells; exsdative pathological changes in the neck of crypt and in villi; and accumulation of mucus. As the result the following were obtained. For computing the mitotic index the method of Leblond was used. 1. Normal controls: The morphology of villi in normal controls may be classified into three; those of normal shape villi (88.5%). those of branching villi (8.4%), and those fused villi (3.0%). Average length of villi proved to be 487±57μ, average width of 140±41μ, and average thickness of mucosa 142±28μ. Histological findings were similar to those reported by other workers, but there could be observed some lymphocytes sparsely scattered in the edithelial cells. In no case, however, there could be detected eosinophils within epithelial cells. Mitotic index was fonnd to be 2.99±0.19. 2. Acute Hepatitis in icteric stage: As for the morphology of villi at the icteric stage in acute hepatitis, some were mixed with short villi, and there were club-shaped villi (13.5%) and mushroom-shaped ones (8.6%). The length of villi tended to be shortened (413±53μ) and in parallel with this shrinkage the mitotic iudex was markedly decreased (1.87±0.59). However, there were no significant differences in the width of villi and the mucosal thickness as compared with those in normal controls. Histological findings revealed disarrangement of epithelial cells, and increase in the number of lymphocytes among epithelial cells, infiltration of lymphocytes and plasmocytes in tunica propria, an increase in goblet cells of villi and crypt, exudative pathological changes in between villi, as well as atrophy and exudative inflammatory changes of duodenal mucosa at the icteric stage in acute hepatitis. 3. At the convalescent stage in acute hepatitis: Morphology of villi approaches to that of normal ones, but there could be obesvred long branching villi (16.7%) and average length of villi tended to increase (537±52μ). Paralleling with these changes, mitotic index increased (3.30±0.52). Histologically, no exudative and inflammatory changes were at all observed in villi and tunica propria.
Pseudomonas aeruginosa produces the two pigments, namely Pyocianine and Fluorescin. The Pyocianine is bluish green pigment and is derivative of phenazine pigment. In this paper, the physiological factors that give influences on the biosynthesis of Pyocianine by Pseudomouas aeruginosa in culture will be presented. 1. Pseudomonas aeruginosa required glycerine as a nitrogene source for the biosynthesis of Pyocianine. In a case that glucose was used as carbon source, the addition of magnesium ion was essential. 2. Within alanine, glutamate and aspartate on which the author examined, only alanine was of effective as nitrogen source for the Pyocianine synthesis. 3. The following results have been investigated by the studies on the atmospheric condition. Pseudomonas aeruginosa produced the pigment at a aerobic condition in the chemically satisfactory culture media, but could not produce at anaerobic condition in any culture media. 4. In the acidic media below pH 6.0, the growing of bacteria was very slight and the Pyocianine could not be produced. Moreover, the pigment did not be synthesized when the pH was descended rapidly, even if the startiag pH of the medium was neutral or slight basic. 5. The growing of the bacteria is indispensable for the synthesis of Pyocianine, then the resting cells could not synthesize the pigment.
92 patients, operated on radical surgery of the cancer of the uterine cervix after the irradiation of telecobalt 2, 000 R. at the Dept. of Obst. & Gynec. Okayama Univ. Med. School, were studied on atypical cells appeared in the peripheral blood (before and after radical operation), in the uterine venous blood and in the wound washings. The cell-collection technique utilizing gum arabic-glucose was followed by the principle of H. Munakata's method. Papanicolaou's staining were applied to all specimens examined. As the result, 13.0% of atypical cells resemble to cancer cells were found in the peripheral blood (before operation), 14.6% in the uterine venous blood, 8.7% in the peripheral blood (after operation) and 25.0% in the wound washings, respectively. The blood vessel invasion of cancer ccells was found to be 29.3% in primary focus. Although the close correlation between blood vessel invasion and lymph node metastasis was found, the correlation between blood vessel invasion and atypical cell appearance could not be observed. A clinical stage or a parametrial infiltration was not correlated with atypical cell appearance. It seemed to be a slight correlation between lymph node metastasis and atypical cells appeared in the circulating blood, however, it was statistically insignificant. A significant correlation between the grade of the lymph node infiltration and atypical cells appeared in the wound washings was fouud. Therfore, it is considerd that the possibility of the dissemination of cancer cells into the peritoneal cavity may occur duriug operative procedures, especially by lymph node resections. On the other hand, a vaginal wall infiltration, a parametrial infiltration or a blood vessel invasion were not correlated with atypical cells appeared in the wound washings. Atypical cells resemble to cancer cells were found in the peritoneal fluid even among benign gynecological diseases. A correlation between atypical cell appearance and the prognosis of the patient is now under investigation.
For the purpose to clarify the influence of respiratory disturbance of the lung in tuberculosis upon the liver, pathological conditions, principally abnormality in the respiratory function of the tuberculous lung, were compared with liver function, selecting 52 cases of lung tuberculosis as the subjects of the study, whose symptoms were relatively steady and without any complication. The degree of the lung function was determined by percent vital capacity. For the examination of the liver function, Gros reaction, Takata's reaction, CCF, TTT. icterus index of Meulengracht, and BSP were undertaken. In the examinations of liver function, the results of Gros reaction, Takata's reaction, CCF, and BSP tended to parallel with the severity of lung tuberculosis, but those of TTT did not reveal any definite relationship, and the icterus index showed only a slight rise in the cases with lung cavitation. As for the relation between the percent vital capacity and the liver function, with an exception of TTT, stoichiometrically there was a significant mutual relation among Gros reaction, Takata's reaction. CCF, icterus index, and BSP, suggesting that the respiratory abnormality of the lung has a great influence upon the liver function.
In order to find out what influence the respiratory functional abnormality would have on urinary lactic acid, pyruvic acid and acetilation rate in lung tnberculosis, the author selected 52 cases of lung tuberculosis whose symptoms were relatively steady and without any complication, and with these cases as the subjects, pathological conditions of the lung, especially respiratory insufficiency as determined by percent vital capacity, and liver functional disorders were compared with lactic acid and pyruvic acid values in urine and acetilation rate. Lactic acid was measured by the hydroquinone method while pyruvic acid by the dinitrophenyl hydrazine method. Acetilation rate was determined by using paraaminobenzoic acid, which was done by Bratton-Marschall's method. Urinary lactic acid was found to be relatively increased only in those cases with lung cavitation, and among the other examinations, especially with the percent vital capacity, there could be recognized no significant relationship. Urinary pyruvic acid was increased in those cases with lung cavitation as well as a mutual relationship with the length of disease. Furthermore, there was observed an aggravating tendency in the lowering of percent vital capacity. As for the rate of acetilation, although there was no definite relationship between the severity of tuberculosis and the percent vital cadacity, all the cases showed a marked decrease in the percent vital capacity, especially so in the groups using PAS and SF. It was thus assumed that chemotherapy has a great influence aside from the pathological conditions of the lung.
After X-ray examinations of a case complaining of upper abdominal pain it was suspected of twin gallbladder, and subsequently it was confirmed by peritoneoscopy. This paper described about this unusual case. It was demonstrated that this case had a twin gallbladder one lobe in front of another each of which measured about 3 cm of diamater. This corresponds to the bilobed gallbladder reported by S. Sherlock. The anterior lobe was hardly functioning and hence the bladder function was performed by the posterior lobe. This point was verified by roentgenographs of the gallbladder as well as by peritoneoscopy.
With the purpose to ascertain clinical significance of urinary mesobilirubinogen in various disease, the author studied by means of the mesobiliviolin reaction changes of mesobilirubinogen contents in the urine of those patients suffering from hepatitis, liver cirrhosis, lung tuberculosis, constipation, and others, who proved positive to the urobilinogen reaction. Simultaneously their liver functions were examined. The results of this study are summarized as follows. 1. It is best to determine the mesobiliviolin reaction six minutes after boiling, and the isolation of mesobiliviolin can be achieved most adequately by column chromatography. By this method there will be yielded some substances other than mesobiliviolin as the by-products. 2. The positive rate to the mesobiliviolin reaction is 50% in liver cirrhosis, 40% in serum hepatitis, and 11.8% in chronic hepatitis, indicating that the severer the injury of liver parenchyma, the higher is the positive rate. In the case of normal persons and those with constipation the reaction is negative. In addition, no positive case can be recognized in those cases with less than 0.40 mg/dl of urobilinogen. 3. There cannot be observed any proportional relation between the urobilinogen concentration and the mesobilirubinogen content. 4. In those cases proved to be positive to the mesobiliviolin reaction the positive rate of various examinations on liver function was 50-62.5%, whereas those positive to urobilinogen it was 15.5%-25.8%. 5. It is assumed that the positivity to urinary mesobilirubinogen reaction is of a great importance for determining the disorder in the liver function.
After inducing experimental liver injury in normal adult rabbits by the injection of chloroform, carbon tetrachloride, or allyl-formate the relationship between the mesobiliviolin reaction at the time when urinary urobilinogen is positive and the results of various examinations on liver function was studied simultaneously, and the following results were obtained. 1. There is found no proportional relation between the concentration of urinary urobilinogen and the positivity of mesobiliviolin reaction, and when urobilinogen is at normal level, the mesobiliviolin reaction proves to be always negative. 2. The histological pictures of the liver when it is mesobiliviolin positive reveal a strong stellate reaction, showing a marked correlation, but there is not necessarily their correlation with the degree of liver cell degeneration. 3. At the time when mesobiliviolin reaction is positive, Takata's reaction and CCF give the rate of 94.1% and 88.2% respectively, showing a marked correlation with one another. TTT gives the rate of 64.4%, being somewhat lower, but there can be observed no correlation between the serum bilirubin level and the positivity to the mesobiliviolin reaction. The degree of BSP injury is 82.3%, showing a good correlation. On the other hand, between the urinary urobilinogen content and these liver functions there is no appreciable correlation. 4. Therefore, even assuming that mesobilirubinogen is disposed of by the liver endothelial system, it represents a more marked injury of the liver than urobilinogen.
In an attempt to know the distribution of Clonorchis sinensis, investigations have been conducted in Chugoku District except for Okayama Prefecture. 1. Parafossalurus manchouricus, the first intermediate host of the fluke, were found in the southern part of Fukuyama City, Hiroshima Prefecture and also in Natajima, Yamaguchi City, Yamaguchi Prefecture. 2. Metacercariae of Clonorchis sinensis were recognized in 6 of 56 (61.5%) Pseudorasbora parva collected and the incidence was significantly high in the southern section of the area. 3. Centrifugal fecal examinations were perforn ed at Natajima and eggs of Clonorchis sinensis were recovered from 12 of 218 (5.5%) residents. Among cats dissected, 4 of 12 (30.0%) were found to be infected with liver flukes.
For different antitumor agents, Nitromin, thio-TEPA, Mitomycin C, and Chromomycin were added in various concentrations into roller tube culture of Bashford carcinoma cells, and thier influences upon the cellular morphology were studied by phase contrast microscopy and by stained preparations. The results obtained are as follows. 1. The minimil growth-inhibiting concentrations of Nitromin, thio-TEPA, Mitomycin C and Chromomycin were 1 r/ml, 0.5 r/ml, 0.01 r/ml and 0.01 r/ml, respectively. 2. The morphological alterations of Bashford carcinoma cells induced these antitumor agents at and around the minimal growth inhibiting concentrations included the following: (a) Nitromin caused dissociation of cells, nuclear alterations and appearance of “inclusion cells” in many cells, (b) thio-TEPA induced rupture of mitochondria and multinucleated giant cells, (c) Mitomycin C was mainly associated with cytoplamic changes such as vacuolization, mitochondrial rounding and cytoplasmic fusion, (d) Chromomycin was destructive to both nucleus and cytoplasm leading to the nuclear pycnosis and rupture of the cell membranes 3. The roller tube culture technique of Bashford carcinoma cells was considered an excellent system for the in vitro screening of antitumor agents.
A spontaneous mammary carcinoma in a high-leukemic AKR strain of mice was studied both in vivo and in vitro, and an attempt was made to elucidate the mechanism of sarcomatous transformation. The tumor, originally a type B adenocarcinoma, transformed into a sarcomatous tumor during serial transplantation in isologous mice. Cell strains were established in vitro from transplants before and after the sarcomatous change. The culture cells were composed of epithelial and flbroblastlike cells and their intermediate forms. It has ceen shown that 1) the carcinoma cells assumcd spindle shapes in loosely growing areas, 2) in the mixed carcinosarcomatous stage, the epithelial tumor cells often merged with the stromal cells, indicating a morphological transition from polygonal to elongated cells, and 3) the behavior and morphology of the culture cell strains were similar, although they were derived from histologically different transplants. These findings suggest that the sarcomatous transformation in a serially transplanted AKR mammary tumor resulted from a morphological alteration of the carcinoma cells.
The fluorocarbon deproteinization technique was demonstrated to be a useful method for the purification of Japanese B encephalitis (JE) virus from infected mouse brain. Electoron microscopic observation on purified virus particles by this method schowed numerous particles which were presumed to be JE virus. Electoron microscopic observation on the surface of the chick red cells which were precipitated with purified virus (hemagglutination) displayed similar particles. The JE particles were hexogonal in schape, and composed of an outer membranes and a centrally located nucleoid-like structure. The size of the particle measured 30 to 40 mμ in diameter. A substance which cause the hemagglutination was conservable to be the virus particle itself.
This report deseribes an investigation of the antigenic localization of Japanese B encephalitis virus in the brain of infected mice that were inoculated intracerebrally with seed virus. 24 hrs. after inoculation, diffuse specific fluoreacence were observed in large cells of the cereberal cortex. In those cells, viral antigen were predominantly present arround the nucleus. In some cells, they spread into the nucleus from the perinuclear region and in others, into the cytoplasm. In case of the brain of mice that manifested typical cereberal signs, fine fluorescent granules were noticed in nerve cells and some glia cell of the cereberum. The majority of the viral antigen appeared to be present in the nucleus. The cells having the viral antigen were most numerous in the region of the substantia nigra, thalamus and basal ganglions. Those cells were also numerous in the cortex and cerebellar cortex. There were no specific fluorescein in the chorioidea and meninges.
Smears of cells of cerebrospinal fluid obtained from patients with Japanese B encephalitis were examined by means of the fluorescent antibody method. Identification of cell types were maid in preparations stained with Giemsa solution after removal of fluorescein-conjugated antibodies. Specific fluorescence was observed in a part of phagocytes, which were the predominant cell type in cerebrospinal fluid in case of Japanese B encephalitis, but no fluorescence in neutrophils. A diffuse distribution of specific fluorescence was seen in the cytoplasm, and its glanular aggregation in a part of the perinuclear region of the phagocytes. Phagocytes schowing specific fluorescence were noticed in all of patients with Japanese B encephalitis and they ranged from 15% to 48% of whole cells seen in preparations. Demonstration of the viral antigen within phagocytes in cerebrospinal fluid by the fluorescent antibody method may contribute to the early diagnosis of Japanese B encephalitis.