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For the purpose to clarify the relationship between production of humoral antibodies and cellular reactions of the lymphoid system to allogeneic-transplanted cells in mice, a study on cross sensitization was carried out between inbred A(H-2a) and C3H(H-2k) strain mice. The median survival time of skin of C3H transplanted to A (C3H-to-A) was 14.1 ± 1. 4 days, and of A transplanted to C3H(A-to-C3H) was 11.8± 1. 6 days. Repeated A cell injections to C3H induced the formation of humoral antibodies, whereas the C3H cell injections into A did not. In A-to-C3H and C3H-to-A combinations, the immunization induced an increase in white blood cell number in circulating blood successively with the repetition of the antigen injection, and organ weights increased in thymus, spleen, and liver but not in kidney. Weight increases in the organs of A treated with C3H cell injection were less in extent, comparing to those of C3H treated with A cells. Histologic observations revealed that the cellular proliferation in the lymphoid system including plasmocytic responses were obviously predominant in the C3H treated with A cells comparing to those in the A treated with C3H cells. Hemocytoblasts also increased during the immunization in both cases showing no significant differences between the two series of experiments. These cellular reactions were observed not only in the draining lymph nodes but also in the generalized lymphoid tissues. The results of the present study suggest that the definitive factor for producing humoral antibodies is in the differences of the homologous antigenicity between the donor and the recipient but not in the degree of sensitization, and the Dk in H-2 loci is not so strong in antigenicity as to elicit sufficient plasmocytic responses for the formation of humoral antibodies in C3H strain mouse.

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.

As a link in the series of studies on tumor-specific immunity, in vitro inhibitory effect of sensitized isologous lymph-node cells on the proliferation of C3H mammary cancer was studied. For this purpose tissue culture was conducted with regional lymph-node cells obtained from truly isologous C3H mouse inoculated with A strain cells derived from C3H mouse mammary cancer along with A cells, and the following results were obtained. In the case of tissue culture with those lymph-node cells obtained from the groups of mice 10 days after the inoculation of 5 X 106 A cells, the inhibitory effect on the proliferation of A cells was most marked, followed by that of those taken on day 14, 7, and 5 decreasing in the order mentioned. In the case with those regional lymph-node cells obtained from mice which did not have recurrence of tumor 1 week after extirpation of 2-week old tumor, the inhibitory effect on proliferation of A cells was marked, with the regional lymph-node cells obtained two weeks after transplantation of 1 × 108 A cells there could be observed no inhibitory effect at all. This suggests that at a certain stage after implantation of such regional lymph- node cells there develops a specific anti-tumor activity in the host.

In the experiments with cultured liver cells it is very important to know whether or not the cells in vitro have the same properties and functions as in vivo. The purposes of this work were to investigate the functions of the cultured liver cells and to identify functionally the liver cells cultured by our present method with the parenchymal liver cells. At first, the albumin production of the cultured liver cells, one of the well known functions of the liver cells, was examined by the immunological methods, especially, the fluorescent antibody technique and the complement fixation test. Culture methods which could display the functions of the liver cells as much as possible were explored simultaneously. The results were as follows: 1. Albumin production was detected in the strain RLN·10 liver cells established from the liver tissues of a Donryu rat with immunofluorescent method and complement fixation test. This confirms that the cultured liver cells maintain the function to produce albumin and these cells have originated from the parenchymal liver cells. 2. Hepatoma strains (AH 66-TC-l, AH 7974-TC-l) also showed the albumin production but the extent of its production was less than that of the strain RLN-10. 3. In the short-term cultured liver cells, the albumin production was testified only slight in one month and was exhibited in a small amount in three months. 4. Every culture method examined exhibited no appreciable difference in the albumin production in the cultured liver cells.

With the purpose to prevent the dissemination and consequent metastasis of cancer cells at the time of operation we gave 10 mg of Mitomycin C per day for four consecutive days prior to surgical operation of gastric cancer (total of 322 patients), and this so-called adjuvant chemotherapy proved to be effective on the cases with serosal involvement and infiltrating type of cancer, irrespective of histological types. It also gave five-year survival rate of 35 per cent. However, to lymph nodes already metastasized, the adjuvant chemotherapy proved to be not effective.

Large doses of adenovirus type 12 were injected intraperitoneally into adult hamsters, and development of tumors and other pathological findings were studied in comparison with those in hamsters injected when newborn. Doses of 38~47 TCID60 per gram body weight produced tumors in 3 of 12 hamsters injected at 37~57 days of age. A dose of 170 TCID60 per gram body weight produced tumors in one of 18 hamsters injected at 61~71 days of age, but in none of 18 hamsters injected at 147~174 days of age, while the same dose per gram body weight produced tumors in 24 of 26 hamsters injected when newborn. In hamsters injected at adult ages, the number of tumors per animal decreased and the latent period for tumor development became very long as compared with those in hamsters injected when newborn. Regardless of the age at the time of injection, acute inflammatory change was observed in the peritoneum which later developed into various degrees of peritoneal adhesion. Adenovirus type 3 also induced the peritoneal adhesion. Histology of tumors was studied and discussed.

DNA synthesis and cell renewal of mouse intestinal epithelium were studied with radioautography after injection of thymidine-H³ to know the difference of the mode of epithelial cell generation relating to the different frequency of cancer developement in several parts of small and large intestines. Succinic dehydrogensase activity was also observed by histochemical method. Renewal time of the intestinal epithelium of mouse is about three days throughout the intestine with somewhat longer time in rectum and anus, and relatively shorter one in ileum compared to the other parts of the intestine. Daily regenerating rate was low in large intestine, especially in rectum and anus. Strong activity of succinic dehydrogenase appeared in the bottom of crypt and seems to be correlated to the active cell division. Epithelial cells in large intestine move very slowly upward and few of them seem to move to the opposite side or stay long time at one place. Intermitotic time is about 27 hours in small intestine and about 40 hours in large intestine. These suggest some relations between the mode of the epthelial cell renewal and cancer development. Because in human the frequency of cancer development is very high in large intestine, rectum and anus, and the epithelial renewal of these areas is supposed to be delayed similarly as in mice.

Histochemical evaluations of human sarcomas such as reticulum cell sarcoma, fibrosarcoma, lymphosarcoma and neurofibrosarcoma, were carried out with five hydrolytic enzymes and eight oxidative enzymes. The activities of acid phosphatase and beta-glucuronidase were slightly positive in the neoplastic cells observed. Beta-esterase activity was also positive but varied according to the kind of sarcomas. Alkaline phosphatase activity was faint or negative in sarcoma cells, though positive in capillary walls. Leucine aminopeptidase activity was negative giving not any appreciable coloration of the cell as far as the method employed is concerned. Among the activities of dehydrogenases, the most intense activity was observed in lactic dehydrogenase. The activities of succinic and beta-hydroxybutyric dehydrogenases were slight. The activities of alpha-glycerophosphate, glutamic and betahydroxybutyric dehydrogenases were faint or slight. The activities of NADPlinked dehydrogenases, glucose-6-phosphate and isocitric dehydrogenase were all faint or slight in these sarcoma cells.

Detailed morphologic characteristics of type C virus particles observed in an X-ray-induced C58 mammary tumor and its transplants have been described. The particles are round and 75 to 100 mμ in diameter, containing an electrondense nucleoid 60 to 70 mμ in diameter. By the negative staining, they do not show obvious spines. Two abnormal types of particles, i. e. cylindrical and aberrant forms have been observed.

Oncogenesis of human adenovirus type 12 in the brain of rats was examined. Newborn rats of Sprague-Dawley and Donryu　strains were injected intracranially with human adenovirus type 12.　The incidence of intracranial tumors was 91% (30/33) in SpragueDawley　and 56% (14/25) in Donryu rats. Except for one tumor nodule　located in the parietal cortex of a Sprague.Dawley rat, all tumors　developed in the paraventricular areas or in the meninges. Tumors were quite similar histologically to those induced in hamsters and　mice resembling the undifferentiated human brain tumors such as medulloblastoma,　ependymoblastoma and embryonic gliomas. From　the histological features and primary sites of tumor development, it is　suggested that the tumors in the brain of rats induced by adenovirus type 12 originate from the embryonic cells in the　paraventricular area　and also from the undifferentiated supporting cells of the peripheral　nerves in the　leptomeninges.

A factor, cornin, inhibiting the growth of L cells cultured in monolayer was extracted from bovine liver with boiling water and was partially purified by gel filtration with Sephadex G-200. The factor was (1) precipitable with ethanol at the concentration between 70% and 90%, (2) impermeable through dializing memo brane, (3) eluted as the last peak at the gel filtration and (4) containing protein and RNA but no DNA.

To analyze the appearance of three forms of hepatitis B antigen-associated particles (HB Ag particles) and antigen-antibody (Ag-Ab) complexes in the sera of patients with various liver diseases, electron microscopic observations with the combinations of a variety of immunological assays were made at first on the HB Ag and Ab mixed in vitro in various ratios, and then on the samples from the sera of each patient. The number of patients observed were 64 in total, which consisted of various types of hepatitis, Hodgkin's disease, Down's syndrome and an asymptomatic carrier. For the detection of HB Ag-Ab complexes a modified method of ALMEIDA was used, and for the isolation of large HB Ag particles (Dane particles) DANE'S method was employed. Electron microscopy proved to be a useful method for detecting HB Ag and the Ag.Ab complexes when the ratio of HB Ag to Ab was in the equivalence. Large aggregates of Ag-Ab complexes were frequently observed in the attacks of acute hepatitis and the recrudescences-of chronic aggressive hepatitis. The aggregates were also observed in fulminant hepatitis but the ratio of HB Ag to Ab was different from each other among 3 cases examined. The large HB Ag particles were not observed in more than half of the cases in the attacks of acute hepatitis, but appeared in the major. ity of cases in chronic aggressive hepatitis, even massively during the period with transiently elevated levels of serum glutamic pyruvic transaminase. A few large particles were also found in sera of an asymptomatic carrier, Hodgkin's disease, and Down's syndrome.

The RNA extracted from Rous sarcoma virus (RSY)induced mouse ascites sarooma cells (SR·C3H, N. P.) by means of the cold SDS-phenol was examined by the electron microscopy on the specimens spread wi th or wi thout urea according to the protein mono· layer technique. The majority of RNA molecules was found in a collapsed agglomerated form, derived from matured ribosomal RNA. Using sucrose gradient, linear molecules of RNA were observed in the interspace of the agglomerated form of RNA at the region of high molecular weight of the band sedimentation. The histogram of the distribution in length of the linear molecules involved up to 6 /1 in length wi th a modal length of 2. 28 f1 and 2.0 to 2. 2 f1 in a pro. minent peak; longer molecules up to 18 f1 in length were scarcely observed. Species of the linear RNA molecules is not exactly known, although this is not mature ribosomal RNA and likely to be messenger RNA or nascent RNA molecules, some of which might associate with RSY·RNA.

Gornin was extracted from bovine liver. The effects of cornin on DNA synthesis were compared with its effects on cell growth using L cells growing in suspension. As the first step of this experiment, a simple method of suspension culture was established with a new modification of YLE medium. Both effects of cornin paralleled with dosage. And the properties of the inhibitory factor of DNA synthesis are the same as those of growth inhibitor in respect to the heat stability and impermeability against dialyzing membrane. The inhibitor of DNA synthesis could not be separated from that of growth by gel filtration with Sephadex G-75.

By injecting 131I-Lipiodol into lymphatics of the dorsum of dog feet, the distribution of 13JI in the lymph nodes and other principal organs as well as its histological effect were studied periodically after the injection for the period of two months. The characteristic feature of J3JI distribution was the fact that J31I was accumulated into lymph nodes markedly higher than in any other organs and it was retained there over a long period of time. Histological examinations of the lymph nodes revealed a marked lymphocytopenia, the loss of germinal center, practically complete loss of lymphoid elements already 5 days after injection, and marked fibrosis. In the lung a considerable J3JI·distribution could be seen in early stage:, but with lapse of time it decreased rapidly. The distribution in other organs such as liver, spleen, bone marrow, kidney, ureter, bladder, thyroid gland, pancreas, testicles and small and large intestines was negligible in amount, and any specific histologic effect of irradiation could not be recognized in these organs including the lung. From these results, the authors concluded that 131I-Lipiodol has a selective activity on lymph nodes by injecting it via lymphatics and it is a safe method in clinical application to treat the patients bearing malignant lymphoma or metastatic lymph nodes.

Firstly, comparisons have been made of the secretion of human growth hormone (HGH) that was induced by insulin, lysine vasopressin and pyrogen injections in order to study whether these substances can be utilized as a rapid test of HGH secretion. In insulin test, a fall of the fasting blood glucose level by 28.6% or more seemed to be sufficient to provoke adequate HGH elevation, and 9.4 ng/ml or higher HGH increment was recognized as being normal, because lysine vasopressin and pyrogen produce varying degrees of side-effects and are less specific and unpredictable in the release of HGH. Secondly, the pharmacologic effects and mechanism of action of exogenous glucagon upon the HGH secretion were studied. In normal subjects after one mg sc glucagon, there was a mean peak blood glucose level of 142. 4±3.l mg/lOO ml at 30 min, HGH levels reached a mean peak level of 22. 6±4. 8 ng/ml at 150 min, and no false negative response was noted. In patients with hypopituitarism, there was no positive response in plasma HGH levels after the sc glucagon. The present study revealed that the rise and subsequent fall of blood glucose are not the sole mechanism responsible for the effct of glucagon on HGH secretion, and that the HGH secretion in response to the sc glucagon was not triggered by cathecholamine via the stimulation of the adrenal medulla.

Ultrastructure of microfilaria Brugia malayi was investigated with electron microscope. Microfilariae are covered by a sheath membrane with dense materials on its outer surfaces. The cuticle consists of 3 layers; namely, external cortical, internal cortical and fibrous layer. Beneath these cuticular layers, thin hypodermis is present and the muscle cells are arranged of 4 groups in a crosssection except for the head and tail. A pair of cephalic channel containing several cilial rods opens at the anterior end of the worm. A hook is situated on the anterior edge of one channel orifice, and several spines grow on the opposite side to the hook. Caudal channels paired laterally opening into the both sides of the posterior region differ from cephalic channels by the presence of a single cilial rod. A central canal runs from the buccal cavity to the inner body, and opens into the inner body cell through the filamentous apparatus. The inner body appears to consist of several cells having storage substances and a flat nucleus located on the periphery of the cell. An excretory apparatus, i. e., a cell, is composed of a nucleus and a large vesicle which has many microprojections on the luminal surfaces. The GI cell which occupies the whole width in a cross-section is larger than the R cell. R2-R4 cells appear to be in a close contact with the anal apparatus having many microprojections on the luminal surfaces. These microprojections differ from those of the excretory vesicle in their thickness and length. The characteristic patterns of these organs are compared with other microfilariae.

Electron microscopic observation was made on the length distibution of messenger RNA molecules in polyribosome pre· paration isolated from mouse ascites sarcoma cells, which was de· stroyed by ethylenediamine tetraacetate treatment in hypotonic solu. tion. The ribosomes appeared first to be a hollowed structure by swelling and then were destroyed to a rod·like structure consisting of ribonucleoprotein strand, which was clearly distinguishable from the linear structure of messenger RNA released from the polyribosomes. The length of messenger RNA was poly.dispersed measuring from 0.02 up to 6 μ, the majority (92%) of which was in the length less than 3 μ with a prominent peak between 0.6 to 0.8 μ.

The appearance of sideroblasts in hypoplastic anemia (HAl and acute myelocytic leukemia (AML), together with their sideroblastograms, was studied. Hematological studies on cases with type III sideroblast dominance by sideroblastograms produced the following results. Type III sideroblast dominant HA was observed in three of 63 cases. Two of the above three cases had what we call "atypical factor", while the remaining one became AML in its clinical course and could be considered to be leukemia in a hypoplastic preleukemic stage. Type III sideroblast dominant AML was noted in five of 32 cases. Three of these five cases are compatible with low percentage leukemia, and one of the above three cases showed ringed sideroblasts exhibiting erythroleukemia in the terminal stage. In HA and AML, type III sideroblast dominant cases have to be examined in relation to atypical HA and atypical leukemia. Changes of iron meta. bolism in erythroblasts with preleukemic stage will be attributable to disturbance of erythropoiesis such as erythroid hyperplasia in bone marrow and also, in part, to disturbance of hemoglobin synthesis.

For the purpose to reveal the changes in stimulatory effect of dibutyryl-cyclic- AMP on erythropoiesis during ontogenetic development, the author studied syntheses of DNA, RNA and protein of erythroid cells in fetal liver, neonatal and adult bone marrows of rats. In the bone marrow of neonatal animals erythropoiesis was stimulated by the intraperitoneal injection of cyclic nucleotide with enhanced DNA, RNA and protein syntheses of erythroid cells. Enhancing effect of dibutyryl-c-AMP on the erythropoiesis decreased gradually with advance of neonatal days. Autoradiographic observations revealed that in erythoid cells isolated from fetal liver and neonatal bone marrows, DNA-, RNA- and protein-snythesis was markedly stimulated by incubating with cyclic nucleotide, but not in those from adult bone marrow. Discussion was made on the changes in the regulatory mechanism of erythropoiesis according to the transition of hematopoietic organs during development.