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MC-induced sarcomas produced under the skin on the back between　scapulas of C3H mice were transplanted　successively to the mice of the　same strain. Using the first and the second generation tumors, viable　tumor cells were prepared and with these tumor cells C3H mice were　inoculated. From these sensitized mice regional lymph　nodes were taken　out at certain intervals and lymph-node cells were prepared. These tumor　cells were mixed with regional lymph-node cells in the ratio of 1 : 10, and the mixed cells were transplanted subcutaneously on the back of C3H　mice, and the development and growth of tumors were observed at intervals.　As a result it was found that the inhibitory effect of these regional　lymph-node cells on the tumor growth was strong one to two weeks after　the transplantation, but beyond 3, or 4 weeks no inhibition　was observable.　In connection with the present in vivo experiments, some comments　were made on the available literature, and it has been demonstrated that　even in the cancer-bearing animal destined to die of tumors, at certain stage of cancer there is seen an inhibitory effect of the host on the tumor　growth by way of the lymphoid system and that such a response of the　host in vivo seems to be　correlated well with in vitro reaction.

For purpose to study specificity in the growth inhibition effect of sensitized lymph-node cells on target cells, the regional lymph-node cells obtained from the truly isologous mouse previously inoculated with A strain cells (derived from C3H mouse mammary cancer) were cultured with A cells in various ways, and obtained the following results. 1. Those regional lymph-node cells from the isologous mice transplanted with the skin of C3H mouse or MC-induced sarcoma do not inhibit the growth of A cells in tissue culture. 2. The regional lymph-node cells from the mice positive to tuberculin test also do not inhibit the growth of A cells in tissue culture. 3. The regional axillary lymph-node cells (C3H anti-A strain cells) inhibit the proliferation of M cells from Cb mouse mammasy cancer and JTC-ll cells from Ehrlich ascites tumor as well as A cells. However, these axillary lymphnode cells do not inhibit the growth of AH-66F cells from rat DAB hepatoma, Hela-S3 cells from human uterine cancer and L cells from subcutaneous connective tissue of C3H mouse. From these results it is assumed that the sensitized regional lymph- node cells act specifically on cancer antigen.

Indocyanine green (ICG) was injected into rat liver nodules induced by 2-acetylaminofluorene (2-AAF) via portal vein. The relationship between ICG staining and cell atypism of liver nodules was examined by means of histology and DNA flow cytometry. After 2-AAF administration, many small nodules appeared on the liver surface. All hyperplastic nodules were ICG stained until 10 weeks after the administration, but some nodules were not stained after 14 weeks. ICG-stained nodules histologically consisted of benign tissues and borderline lesions, and many of them showed "diploidy" in DNA cytometry. ICG-unstained nodules consisted of hepatocellular carcinoma (HCCs) and borderline lesions, and many of them showed "aneuploidy". In this way, it has been suggested that HCC could derive from hyperplastic nodules and that they might lose an ability to take up ICG in the process of hepatocarcinogenesis. Immunohistochemical staining for glutathione-S-transferase alpha (GST-alpha), a carrier protein of ICG in hepatocytes, was well correlated with ICG staining in the nodules, suggesting that the loss of ICG uptake in HCC was partly due to the decrease of GST-alpha. Moreover, the appearance of ICG unstained and aneuploid nodules was significantly inhibited in rats which were fed on diet containing Syosaiko-to after the administration of 2-AAF. Chemopreventive effect of Syo-saiko-to on hepatocarcinogenesis was identified.

1. Addition of nicotinamide (l0-2M) into the culture medium brings about an increase of the NAD content and the inhibition of the growth of L cells in culture. This rise of NAD brought about by nicotinamide lasts for 2 to 3 days, and thereafter gradually subsiding, it returns to normal level. 2. When L cells are cultured for several days in the same medium without addition of nicotinamide, there occurs a slow-down of mitosis with lapse of cultivation time but it has been found that this is in no way connected with the intracellular content of NAD. 3. By the addition of isonicotinic acid hydrazide (l0-2M) into the culture medium, there can be recognized a decrease of NAD content in L cell and the inhibition of cell growth. 4. In the case when 3-acetylpyridine (l0-2M) is added, a decrease of intracellular content of NAD in L cells and a marked inhibition of the cell growth can be observed. In the groups cultured in the media, containing 3-AP at the concentration of l0-3M or l0-4M can be seen neither inhibition nor acceleration of the cell growth. The oxygen uptake of the cells cultured in the medium containing 3-AP (l0-2M) hardly differs from that of the control group cultured in the medium not containing 3-AP. 5. On the basis of these results discussion has been made on the relation ship between mitosis and NAD content in the 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.

In the mixed tissue culture of mouse lymphocytes with addition of PHA the rate of the appearance of large and intermediate cells increases markedly, but which side of the two cell groups have reacted stronger remains obscure. In order to solve this problem, mixed cultures were conducted in such a way that only one cell group of the two would react. Namely, one cell group was exposed to C0 6).irradiation (Table 2) prior to the culture and cultured with another viable cell group (FJ test group, Table 2) to see the percentage of the appearance of large and intermediate cells. Simultaneously, the skin homograft from respective donor mouse was transplanted to each other and the survival days of each skin graft were compared. As a result it has been shown that the percentage of blastformation and the survival time of the skin transplant in each group prove to be in an inverse relation. The results of these mixed cultures indicate that the extent of blast. formation reflects significantly the difference in B-2 histocompatibility antigens.

1. It has been found that mouse lymph.node cells, even destroyed by sonication with 20 KC supersonicator, maintain sufficient antigenicity both in vitro and in vivo. 2. When such sonicated cell homogenate is cultured with live lymph-node cells, there can be observed blastformation and the peak of the rate of the blastformation is seen at culture hour 48. 3. When PHA (phytohemagglutinin)-M is added to such mixed cultures, the blastformation is enhanced. 4. When mixed cultures of mouse lymph-node cells are conducted by using such one-way stimulation method in various combinations, the rate of blastformation can tell quite accurately the differences in H-2 antigens of mice. 5. In the experiment using F1 hybrid mice and the parents, it has been demonstrated that the rate of blastformation in mixed cultures of the present experiments shows a direct correlation to the rate of blast formation in mixed cultures of live lymph node cells, whlie it is an inverse proportion to the survival time of the skin transplant. 6. Differences in the transplanation antigens said to be located on sex chromosomes cannot be distinguished by this one.way stimulation method.

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.

1. An oligomycin -sensitive ATPase was isolated and partially purified from beef heart mitochondria. The specific activity of ATPase sensitive to oligomycin of the fraction was five to eight times that of aged mitochondrial or of DNP-induced mitochondrial ATPase assayed under the same condition. 2. Electron micrographs of the partially purified oligomycin- sensitive ATPase reveal a structure in which headpieces are regularly attached by way of stalks to a thread-like structure derived from a superficial portion of base pieces. 3. A high concentration of the structured material coincided with a high activity of oligomycin-sensitive ATPase. When the headpieces were detached from the structure, the ATPase became insensitive to oligomycin. 4. The fraction of oligomycin -sensitive ATPase was essentially free of membrane structure and was contaminated with a small amount of cytochromes b and Cl but no cyt. a. Cytochrome concentrations of the preparations were indifferent to the activity of oligomycin sensitive ATPase. It follows that ATPase does not require cytochromes or membrane structure for its oligomycin sensitivity. 5. From these results it seems that the factor rendering ATPase sensitive to oligomycin should be contained in the stalks and/or the thread-like portion of basepieces of the structure. The structure is the simplest unit of oligomycinsensitive ATPase as yet obtained. 6. The structure was called "oligomycin-sensitive ATPase particles" (abbreviated as OSA particles). A unit of OSA particles consists of a headpiece attached by a stalk to a portion of base piece.

We developed an indirect capillary tube method to improve reproducibility of macrophage migration inhibition (MI) tests using a one-way mixed lymphocyte culture. MI response could be induced to cell-surface antigens coded by either H-2 or non-H-2 (background) genes. The sensitivity was more readily induced across H-2 + background differences. The presence of only background difference did not induce the MI response to much extent. High MI activities were obtained to antigens coded by either K end or D end of the H-2 complex + background difference. Moderate activities were induced across the H-2D difference + background. These results suggest that the D region of the H-2 complex may direct a MI response when an H-2I difference is present during sensitization.

Epstein-Barr virus (EBV)-related herpesvirus (Si-IIA-EBV) was serially transmitted for 3 passages from rabbit to rabbit of the opposite sex by blood transfusion, which subsequently induced virus-associated rabbit lymphomas. The virus could be transmitted by transfusion with 15-20 ml of whole blood (7/7) or irradiated blood (1/6) from the EBV-related virus-infected rabbits, but there was no transmission with transfusion of cell-free plasma (0/6) from the infected rabbits. Passive anti-EBV-VCA IgG (x 20 approximately x 10) titers decreased during the first 1-2 weeks in the transfused rabbits. The virus-transmitted rabbits showed a gradual increase in antibody titers ranging from peak titers of x 640 to x 2560 after 3 weeks of transfusion. The recipient origin of malignant lymphoma that developed in the first rabbit transfused by infected blood was confirmed by chromosomal analysis. This rabbit model thus shows that EBV-related herpesvirus is serially transmissible by blood transfusion and that transmission can not be completely prevented by irradiation of blood, but removal of blood cells is the best way to prevent transmission of EBV-related virus. Therefore, this animal model provides a convenient in vivo system for studies of the prevention and therapy of transfusion-related transmission of EBV and EBV-associated lymphoproliferative diseases in immunocompromised human beings.

1. Adrenalin, when acting directly on eosinophils, brings about a diminution in the wandering velocity of eosinophils but it has no influence on the number of the cells. Judging from the movement patterns of eosinophils this drug acts as to impede the motive function. 2. Acting directly on eosinophils, cortisone markedly decreases the wandering velocity of these cells and also brings about the diminution in the number of the cells. Likewise from the movement patterns of eosinophils, this drug markedly impedes the motive function of the cells. 3. ACTH (adrenocorticotropic hormone), when acting directly on eosinophils, enhances the wandering velocity of eosinophils but it in no way affects the number of eosinophils. Judging from the eosinophil movement patterns, this drug markedly promotes the motive function. 4. Although adrenalin brings about a decrease in the number of peripheral eosinophils in hypophysectomized dogs, the rate of the decrease is less than that observable in the case of normal dogs. 5. Cortisone brings about no significant change in the number of peripheral eosinophils in hypophysectomized dogs, but is induces a decrease in peripheral eosinophils of normal dogs. 6. ACTH acts as to decrease the number of peripheral eosinophils to an equal. degree in both hypophysectomized and normal dogs. 7. When cortisone is administered simultaneously with Adrex, a marked decrease in peripheral eosinophils is brought about in hypophysectomized dogs. 8. By means of the bone-marrow tissue culture of hypophysectomized dogs it has been confirmed that the blood plasma of hypophysectomized dogs lacks an essential factor for cortisone to induce eosinopenia in perpheral blood. 9. The decrease in eosinophils of peripheral blood induced by cortisone has been proved to be dependent upon the presence or absence of the pituitary body. 10. Taking the decrease in peripheral eosinophils by cortisone administration as the criterion, the author has carried out clinical observations with this method and obtained anticipated results.

Sporadic intestinal cryptosporidiosis is not easily diagnosed and might be overlooked. We present here a case of this disease in a 23-year-old Japanese military man with 3 days of abdominal pain, watery diarrhea, and nausea. The frequency of his diarrhea was more than 10 times per day. After his diarrheal bowel symptoms subsided, a colonoscopy was performed because inflammatory bowel disease was suspected. Although the endoscopic findings indicated non-specific ileitis, intestinal cryptosporidiosis was suspected from the histology of ileal biopsy specimens, and this was confirmed ultrastructurally. At that time, however, the patient was on active duty, and thus it was not possible to confirm this as a definitive diagnosis by an adequate stool examination for cryptosporidium. Routine practitioners should be encouraged to carefully inspect patients for this disease, supported by detailed knowledge of it and its diagnosis. If stool-examination results are negative or are not obtained at first, histological diagnosis by endoscopic biopsy could be a useful way to screen for intestinal cryptosporidiosis. Furthermore, stool or histological examination should be performed in recovered patients because the oocysts may continue to be shed for 1 to 4 weeks after the symptoms disappear. Therefore, endoscopic and histological examinations may be useful tools for the early diagnosis of intestinal cryptosporidiosis, although admittedly they are invasive procedures.

The existence of autonomic adjustment functions of eye pressure was demonstrated in various ways, both clinically and experimentally. It is possible to consider that in normal condition, I.O.P. is controlled autonomically like cardiac or respiratory rate irrespective of the internal or external influences of the body. The auther calls such a phenomenon "autonomic eye pressure adjustment function". The mechanism of this physiological function will be reported on in articles to follow.

From the results of various experiments in an attempt to investigate the relationship between the intraocular pressure and the ophthalmic nerve, the author has come to the conclusion that the ophathalmic nerve is one of the afferent pathways transmitting the various impulses caused by the changes in the intraocular pressure to the autonomic eye pressure center, and the impulses created by these stimuli in the eye pressure center are in turn transmitted to the eyeball by way of the autonomic nerves and thus the eye pressure is autonomically regulated by these reflexes.

We evaluated the long-term outcome of 148 patients with small cell lung cancer (SCLC) who had been entered into clinical trials of chemotherapy with or without thoracic and prophylactic cranial irradiation (PCI) between 1981 and 1987. Eighteen patients (12%) survived for 2 or more years. With a minimum follow-up of 4.5 years, 10 of the 18 patients who remained disease-free at 2 years are currently alive and free of SCLC. Seven of these 10 patients currently function as they did before diagnosis. However, three suffer from central nervous system changes of varying degrees in severity which appeared 2-3 years after PCI. Eight of the 18 patients who were disease-free at 2 years have died. Two died of isolated relapse in the brain at 3.6 and 4.2 years after initiation of chemotherapy. Five died of other malignancies while continuing their complete response to SCLC; two of non-small cell lung cancer, two of acute myelogenous leukemia, and one of hepatocellular carcinoma. Another patient died of an unrelated disease without any evidence of SCLC. A small but substantial proportion of patients who underwent intensive treatment will achieve long-term survival; however, these patients remain at higher risk for second cancers and late toxicities. Therefore, attention must be directed to defining the safest way to employ such treatment in the management of SCLC.

The in vivo effects of human placental extract (1-4 ml/kg) on hepatic lipid peroxidation, blood and liver glutathione (GSH) levels and several enzymes associated with the antioxidant defence mechanism; i.e., catalase, glutathione peroxidase, glutathione reductase and glutathione S-transferase, together with some blood biochemical responses were investigated in rats. At an optimal dose level (4 ml/kg), a single acute intraperitoneal administration of the extract caused a significant enhancement (49.9%; p < 0.001) of lipid peroxidation with a decline in GSH level both in blood (45.1%; p < 0.001) and liver (61.0%; p < 0.001) in comparison to control animals. Activities of catalase, glutathione peroxidase and glutathione reductase were inhibited in a dose-responsive way by the treatment with the extract which also increased the activity of glutathione S-transferase in a dose-dependent manner. The extract was found to be hepatotoxic in terms of elevation of serum glutamate oxaloacetate transaminase, serum glutamate pyruvate transaminase, serum lactate dehydrogenase and blood methemoglobin concentration. Results of this study suggest the adverse consequences of the administration of the extract due to its substantial ability to alter normal cellular processes.

A high performance liquid chromatographic (HPLC) method with both electrochemical detection (ECD) and ultraviolet spectrometric detection (UVD) was developed for the rapid and simultaneous measurement of estradiol (E2), estrone (E1), testosterone (T), 17 alpha-hydroxyprogesterone (17-OHP) and progesterone (P) in serum. These hormones were extracted with diethylether, and chromatographed on an octadecyl silane-silica (ODS) column with an eluent of a phosphate buffer solution - acetonitrile mixture (volume ratio 49:51). Estrogens were detected by ECD at +1.0 V vs. Ag/AgCl, and other hormones by UVD at 242 nm. With this method, the simultaneous determination of sex steroid hormones could be performed within approximately two hours with high precision. The hormones of 34 patients (39 menstrual cycles) undergoing human menopausal gonadotropin (HMG)-human chorionic gonadotropin (HCG) therapy were measured. It was concluded that the switch from HMG to HCG should be performed when the E2 level reaches 400 pg/ml for ovulation and 800 pg/ml for pregnancy. The occurrence of ovarian hyperstimulation syndrome (OHSS) can be predicted when the P level rises above 30 ng/ml on the 7th day after the switch. Moreover, conception may be indicated when the P level does not increase from the 7th to 14th day after the switch. In this way, this method proved to be useful for the monitoring of HMG-HCG therapy.

With the purpose to see if GABOB is in any way concerned with the mechanism of the epileptic attack observations were carried on the oxygen consumption of the brain homogenates of rabbits, normal and CLA, and of human, epileptic and non-epileptic. The experiment proved that the oxygen consumption is increased in the epileptic brain and in the brain of CLA rabbit. It was raised by adding ATP-Na salt or DPN, but GABOB itself showed only a slight effect. The results suggested that the oxygen consumption of brain is not so closely correlated with GABOB, but there is a possibility that the decrease in GABOB contents in epileptic brain by the accelerated decomposition with its elevated oxygen consumption may be correlated to the epileptic attack, though the final conclusion requires further observations.