Okayama Igakkai Zasshi (Journal of Okayama Medical Association)

Electron Microscopic Studies on Oncogenic Virus

II. An electron microscopic Study on the virus particles of spontaneous mammary carcinoma in the strain C3H mice

Kazuhiko IKEDA

1966 Volume 78 Issue 6 Pages 655-664
Published: June 30, 1966
Released on J-STAGE: February 13, 2009

DOIhttps://doi.org/10.4044/joma1947.78.6_655

JOURNAL FREE ACCESS

Show abstractHide abstract

Spontaneous mammary carcinoma of C3H mice was studied by electron microscopy.
Cancer cells showed prominent nucleoli and increased nucleocytoplasmic ratio, containing a few mitochondria, vacuoles, and endoplasmic reticulum in the cytoplam. There were inclusion bodies in some of cancer cells.
Mature virus particles were seen in the lumina of the milk ducts, intercellular space and intracytoplasmic vacuoles. They were round in shape and composed of an eccentrically-located, electron-dense nucleoid containd in a triple-layered shell ranging 100 to 120 mμ in diameter. It was demonstrated that the outermost layer of the shell consists of numerons 100 Å spines. This finding is closely consistent with the negative contrast figure of the mature virus particles.
A new interpretation of type A particles was also presented.

View full abstract

Download PDF (2358K)
Studies on the role of virus in the development of leukemia by X-ray-irradiation

Report 1. Serial cell-free passages of X-ray-induced RF mouse leukemia

Naoko OKADA

1966 Volume 78 Issue 6 Pages 665-675
Published: June 30, 1966
Released on J-STAGE: February 13, 2009

DOIhttps://doi.org/10.4044/joma1947.78.6_665

JOURNAL FREE ACCESS

Show abstractHide abstract

In an attempt to study the role played by the virus in leukemia induced by X-ray irradiation, the author described the successful serial passage of the cell-free filtrate from RF mice with leukemia by X-ray irradiaion.
Cell-free filtrate from RF mice with leukemia induced by exposure of the entire body to a single dose of X-ray was bioassayed by the inoculation to new-born RF mice within 16 hours of birth. Four of sixteen test mice developed leukemia in 357 days, 374 days, 616 days and 663 days respectively. Serial implantation of the cell-free filtrate was successful in 60 per cent, 6 of 10 test mice, on the second passage, and in 17 per cent, one of 6 test mice, on the third passage. No leukemia was induced by the inoculation of cell-free filtrate from normal RF mice.

View full abstract

Download PDF (2212K)
On structure of flagella of El Tor vibroio

Jutaro TAWARA

1966 Volume 78 Issue 6 Pages 677-682
Published: June 30, 1966
Released on J-STAGE: February 13, 2009

DOIhttps://doi.org/10.4044/joma1947.78.6_677

JOURNAL FREE ACCESS

Show abstractHide abstract

In this experiment it was possible to observe structure of the flagella of El Tor vibrio. Namely, in the observation of the flagella of El Tor vibrio being cultured on alkaline nutrient agar there can be observed a sheath surrounding the core, but morphologically it is quite unstable, revealing swollen sheathed parts due to autolysis and nonsheathed parts exposing the core. It has also be clarified that two kinds of the flagella, big and slender, sometimes observable are the big ones with intact sheath and the smaller ones that have lost their sheath and the core being exposed. The core in sometime is found enlarged in places showing bumps in several portions but it has been demonstrated to be morphologically very stable.

View full abstract

Download PDF (1298K)
Immunofluorescent Studies of Viral Antigens in Mouse Leukemias

I Purification of Viral Antigens in C₅₈ Mouse Leukemias by Fluorocarbon Extraction

Yoshiaki TAKAHASHI

1966 Volume 78 Issue 6 Pages 683-694
Published: June 30, 1966
Released on J-STAGE: February 13, 2009

DOIhttps://doi.org/10.4044/joma1947.78.6_683

JOURNAL FREE ACCESS

Show abstractHide abstract

Purification of viruses of C₅₈ mouse leukemias was carried out by the following procedures; filtration, differential ultracentrifugation and Gessler's fluorocarbon extraction. The degree of virus purification was examined by fluorescent antibody method, electron microscopy, nucleic acid analysis and bioassay.
The following results were obtained.
1. Large amount of host cell antigens was noted in virus suspension separated by Berkefeld-N and Chamberland L₃ filtration and by differential ultracentrifugation. Therefore it was improper to perform immunofluorescent study because this procedure was concerned with examination of viral antigens.
2. Immunofluorescent procedure by Coons' direct method using anti-viral serum against fluarocarbon extracts showed partially granular or partially diffuse specific fluorescence in leukemic cells and intercellular spaces of the leukemic organs.
3. Electron microscopic observation revealed many virus-like particles in the fluorocarbon extracts. And 50% of animals inoculated with developed leukemias in these extracts.
4. Ultraviolet spectrophotometry of the fluorocarbon extracts demonstrated a maximal absorption at 260 mμ and a minimal absorption at 230 mμ with a Bechman spectrophotometer. It was clear from these data that large amount of nucleic acid was contained in these extracts. And also it was indicated by Bial's test that the fluorocarbon extracts of the leukemia organs contained large amount of RNA.
5. From above findings, fluorocarbon extraction appeared a superior procedure for fluorescent antibody technic in comparison with the other procedurs used. It was thought that the specific fluorescence in the immunofluorescent study represented viral antigen itself.

View full abstract

Download PDF (957K)
Immunofluorescent Studies of Viral Antigens in Mouse Leukemias

II Organ Distribution and Intracellular localization of Viral Antigens in C₅₈ Mouse Leukemias as studied with Immunofluorescent complement fixation method

Yoshiaki TAKAHASHI

1966 Volume 78 Issue 6 Pages 695-705
Published: June 30, 1966
Released on J-STAGE: February 13, 2009

DOIhttps://doi.org/10.4044/joma1947.78.6_695

JOURNAL FREE ACCESS

Show abstractHide abstract

Organ distribution and intracellular localization of viral antigens in C₅₈ mouse leukemias were investigated in detail by the immunofluorescent complement fixation method. During successive transmission of C₅₈ mouse lymphocytic leukemia by cell-free filtrates myelocytic leukemia developed. Whether this myelocytic leukemia was caused by the same or different virus was investigated by the above immunofluorescent procedure.
1. More distinct and increased particle fluorescence which indicated the presence of leukemia viral antigens were detected by the immunofluorescent complement fixation method than by the direct method.
2. The viral antigens in the leukemic cells were localized in 4 different types; perinuclear arrangement, intracytoplasmic scattering, arrangement near the cytoplasmic membrane and extracellular localization. There was no observable difference in localization of viral antigens between the lymphocytic and myelocytic leukemias.
3. Of organ distribution, more viral antigens were observed in the bone marrow and lymph nodes and a few in the spleen and liver in the lymphocytic leukemia. Less viral antigens were observed in the bone marrow and lymph nodes in the myelocytic leukemia.
There were no viral antigens in the organs of the non-leukemic C₅₈ mouse. Cross-immune reactions using antiviral antisera of the lymphocytic and myelocytic leukemias showed that the two viruses closely resembled but were not identical. These findings were indicative of the presence of the two types of viruses in the C₅₈ mouse leukemias.

View full abstract

Download PDF (957K)
The cytochemical studies on the tissue culture cells infected with measles virus and distemper virus

Part I. Study of the inclusion bodies in the tissue culture cells of measles virus infection

Jun SHIBATA

1966 Volume 78 Issue 6 Pages 707-719
Published: June 30, 1966
Released on J-STAGE: February 13, 2009

DOIhttps://doi.org/10.4044/joma1947.78.6_707

JOURNAL FREE ACCESS

Show abstractHide abstract

The tissue culture cells, dog kidney, HeLa-S₃ and PS cells infected with measles virus (Edmonston strain) were studied with the staining methods of Feulgen reaction, methylgreen pyronine and acridine orange, and with RNase digetion test. The results were as follows.
1) The CPE of the cultured cells inoculating measles virus was observed as many authors reported to be consisted of the multinucleated giants cells, cytoplasmic inclusion bodies, intranuclear inclusion bodies and spindel like cells,
2) The cytoplasmic inclusion bodies showed Feulgen negative, red staining with methylgreen pyronine, pale blue to pale green with acridine orange and RNase resistent. As a result presumably they had RNA, but did not always neglect the presence of acid mucopolysacchrides.
3) The intranuclear inclusion bodies had no nucleic acids because no fluorescence was detected corresponding to most of them.
4) With acridine orange staining, the nucleoli of the infected cells became enlarged and showed brilliant red fluorescence, and there was an increase in the size and yellow fluorescence in the nuclei. Cytoplasmic red fluorescence also increased. It seemed to be an accerated metabolism of nucleic acids in the infected cells.
5) The small brilliant red fluorescent particles with halo in the cytoplasm were basophylic when restained with hematoxylin eosin. Therefore they were not identified with the cytoplasmic inclusion bodies.
6) It was not determined whether the cytoplasmic inclusion bodies and intranuclear inclusion bodies were the sites of the synthesis of viral component or not with the methods of nucleic acid staining. But they presumably related to the synthesis of viral components.

View full abstract

Download PDF (1981K)
The cytochemical studies on the tissue culture cells infected with measles virus and distemper virus

Part II. A comparative study on dog kidney cultured cells of measles virus and distemper virus infection

Jun SHIBATA

1966 Volume 78 Issue 6 Pages 721-731
Published: June 30, 1966
Released on J-STAGE: February 13, 2009

DOIhttps://doi.org/10.4044/joma1947.78.6_721

JOURNAL FREE ACCESS

Show abstractHide abstract

The morphological changes observed in dog kidney cultured cells infected with distemper virus (Hokkaido strain) were compared cytochemically with those of measlea virus (Edmonston strain). The results were as follows.
1) Twelve to fourteen after inoculation the CPE was first observed in dog kidney cells infected with distemper virus. The appearanoe of the CPE was seven to ten days later than that of measles virus.
2) The CPE of the distemper virus infected cells was the same multinucleated giant cells, cytoplasmic inclusion bodies, and intranuclear inolusion bodies as those observed in the measles virus infected cell.
3) Both the inclusion bodies in the distemper virus infected cells also showed Feuglen negative, red staining with methylgreen pyronine, RNase resistent, and the cytoplasmic inclusion bodies blue to bluish green, otherwise the intranuclear inclusion bodies unstainable with acridine orange. This indicates that as with measles virus infection the cytoplasmic inclusion bodies have RNA, acid mucopolysacchrides and basic proteins, the intranuclear inclusion bodies no nucleic acids.
4) In the tissue culture cells inoculated with distemper virus the intranuclear inclusion bodies appeared at the same time or a few days earlier than the cytoplasmic inclusion bodies when stained with Giemsa or acridine orange.
5) The red particles in the cytoplasma of the measles virus infected cells were not detected in the cells inoculated with distemper virus when stained with acridine orange.
6) With acridine orange staining there were no differences of largeness and brightness of the nuclei and nucleoli between the distemper virus infected cells and the control cells. Then it seemed seldom to be an increased metabolism of nucleic acids.
7) Summing up, the morphorogical changes in the tissue culture cells infected with measles virus and distemper virus were quite the same except for the appearance of the CPE after virus inoculation.

View full abstract

Download PDF (1918K)
Effects of Radioisotopes on Normal Rabbits

1. Effects of internal irradiation normal rabbits after the injection of ⁵⁹Fe

Michio YAMAMOTO, Masatada TANABE, Naomi KATSUMATA, Yoshihiro HADA

1966 Volume 78 Issue 6 Pages 733-739
Published: June 30, 1966
Released on J-STAGE: February 13, 2009

DOIhttps://doi.org/10.4044/joma1947.78.6_733

JOURNAL FREE ACCESS

Show abstractHide abstract

As a step for the measurement of the reticuloendothelial systems (RES) functions, there is a method of labeling serum iron colloid with ⁵⁹Fe. Employing the same method, this study was conducted in an attempt to see whether there would be any differences in the radiation effects when ⁵⁹Fe is injected at varying doses.
Experimentals: The animals used were healthy normal rabbits and they were divided into different groups of those receiveng the doses of 10μCi, 50μCi, 250μCi, and 700μCi as well as those injectcd with non-radioactive serum iron colloid solution, and simultaneously congored was injected to every group and then the congored indices were obtained. One week later identically the same doses for the respective groups were again injected. and also congored, and the congored indices of these two trials were compared. Fluctuations of the ⁵⁹Fe level in whole blood were observed. The animals were sacrificed by exsanguination at 12 hours and 96 hours after the injection. On the tenth day after the injection hemograms were studied to determine the effects of the injections.
Results: 1. It has been found that the higher the dose of ⁵⁹Fe, the greater is the inhibitory effect on the ⁵⁹Fe incorporation into hemoglobin, but the dose of ⁵⁹Fe itself seems to have little effect on the function of phagocytosis.
2. There can be observed no appreciable differences in the hemograms on the tenth day after the injection among these groups of animals.
3. The quantity of iron deposited in various organs at 12 hours and 96 hours after the injection decreased in the order of bone marrow, liver and spleen in all the groups receiving ⁵⁹Fe but there could be seen no differences among these various groups.

View full abstract

Download PDF (820K)
Histochemical Studies on Carbonic Anhydrase

1. Observation of carbonic anhydrase activity in endometrium

Takashi KUSUMOTO

1966 Volume 78 Issue 6 Pages 741-750
Published: June 30, 1966
Released on J-STAGE: March 31, 2009

DOIhttps://doi.org/10.4044/joma1947.78.6_741

JOURNAL FREE ACCESS

Show abstractHide abstract

The present communication describes the results of histochemical observations made on the localization of carbonic anhydrase in the mucosa of uterus and its tubes. The effect of hormones on this enzyme is also reported. The human, rabit and rat endometrium are used as materials in this experiments.
1. It has confirmed that a modification of Hausler's method is an excellent one for the histochemical demonstration of carbonic anhydrase.
2. In the human endometrium the activity of carbonic anhydrase is found to fluctuate with the menstrual cycle; namely, it increases during the secretory phase, while it decreases markedly or disappear at menopause.
3. Hisochemical demonstration of carbonic anhydrase in the human endometrium is obtained in the cytoplasm of the cells of endometrial gland and surface epithelium if the endometrium.
4. In the mucosa of human uterine oviduct, the activity of carbonic anhydrase is found, but it is rather stronger in the subserosal portions.
5. In the endometrium of young rabbit, the activity of carbonic anhydrase cannot be observed, but on priming with estrogen followed by progesterone administration, the enzyme activity appears. In the case of adult rabbits, while the activity cannot be detected without treatment, it can be recognized after the administration of progesterone alone without the priming the with estrogen.
6. In the adult rat endometrium, the carbonic anhydrase activity cannot be detected in any stage of the sexual cycle. In the group after mating the enzyme activity is demonstrated in every case and it's localization is found to be highest in the glandular epithelium of endometrium. In the subserosal tissues of the uterus and decidual changed portions, the enzyme activity slightly can be seen.
7. From these findings, carbonic anhybrase activity seems to be associated with the nidation and the growth of fertilized ovum.

View full abstract

Download PDF (1228K)
Histochemical Studies on Carbonic Anhydrase

2. Enzyme activity in cultured cells

Takashi KUSUMOTO

1966 Volume 78 Issue 6 Pages 751-756
Published: June 30, 1966
Released on J-STAGE: February 13, 2009

DOIhttps://doi.org/10.4044/joma1947.78.6_751

JOURNAL FREE ACCESS

Show abstractHide abstract

In the present experiment, histochemical study of carbonic anhydrase was made with the use of various strain cells and following results were obtained.
1. By a slight change in the components of Haüsler's substrate a technique has been established, with which it is possible to demonstrate the localization of carbonic anhydrase in the cells.
2. The localization of carbonic anhydrase has been demonstrated in various strains cell such as HeLa cells, JTC-3 cells, and L-cells. Further, it has been found that this enzyme is localized in the nucleolus and cytoplasm in evey strain cells and the activity of the enzyme is highest in the nucleolus.
3. In the observation conducted with HeLa cells to see the effect of hormones on the enzyme activity, it has been observed that the addition of estrogen inhibits the activity carbonic anhydrase but by the further addition of progesterone the activity can be restored. However, there can be recognized no change in the enzyme activity on the addition of progesterone alone.

View full abstract

Download PDF (1248K)
[title in Japanese]

1966 Volume 78 Issue 6 Pages 757-763
Published: June 30, 1966
Released on J-STAGE: February 13, 2009

DOIhttps://doi.org/10.4044/joma1947.78.6_757

JOURNAL FREE ACCESS

Download PDF (419K)

Register with J-STAGE for free!