An electron microscope study of liver cell in carbon tetrachloride intoxication, significance of "opaque area"

Electron microscope study on the rat liver cells of carbon tetrachloride poisoning has been reported. Observations have been made on the osmic fixed tissue sections obtained from the liver at early stages of poisoning, 5 to 22 hours after carbon tetrachloride oral administration, 0.25ml. per 100g. body weight. Special attention is paid on the appearance of electron dense area, opaque area, in cytoplasm, which is compased of fibrous components, probably originated from endoplasmic reticulum. This will be an important sign of cell degeneration. Toluidine blue, PAS and methyl green-pyronin stainings of the thicker sections from the same samples as used for electron microscopy revealed that the opaque area is stained by toluidine blue and pyronin but not by PAS. The opaque areas appear already five hours after the carbon tetrachloride administration and show some continuity with elongated filaments of endoplasmic reticulum. At an advanced stage of paisoning the opaque area increases in its number and size, but some of them are shrunk as a mass, being separated from the surrounding cytoplasm with scanty area. Often they form denser masses in the center and loak like the lipid deposition. The picture suggests formation of lipid droplets in the case of fatty degeneration of the liver cell.</p

Growth inhibition and morphologic changes of HeLa cells exposed to unsaturated fatty acid fraction from the liver of x-ray irradiated rabbits (OX)

Biological effect of the unsaturated fatty acid fraction from the X-ray irradiated rabbit liver (OX) on HeLa cells has been observed in vitro comparing with the effect displayed on the same strain cells by the unsaturated fatty acid fraction from the non-irradiated rabbits, which is extracted by the same method as in OX, The observations have proven that OX is a powerful cytotoxin in a concentration of 0.05-0.025 per cent and induces a severe cell degeneration and cell death, resulting in a marked arrest in the growth of the cells. The similar effect has been observed by unsaturated fatty acid fraction from the nonirradiated rabbits, but the effect was much less comparing to that of OX. Possible mechanism of the cell damage by OX has been discussed.</p

Chemical analysis and biological activities of fatty acids from the liver of x-ray irradiated rabbit, the antitumor agent so-called OX

Chemical and biological characteristics of the unsaturated fatty acids from the liver of irradiated and non-irradiated animals and some unsaturated fatty acids in sale have been described. The unsaturated fatty acid fractions from the rabbit liver taken after irradiating animal with x-ray show hardly any difference from those of non-irradiated animal in each component. But the former were distinguished from the latter in the increased rate of velocity of autoxidation. Similar characteristics were observed on the unsaturated fatty acids irradiated in vitro. They developed less labile free radicals with the shift of the double bonds to the carboxylic group and the conjugated double bonds, dienoic and trienoic acids. Biologically, the fatty acids from the irradiated animal suppressed the growth of bacteria requiring unsatturated fatty acid. And they are slightly stronger in the activity of uncoupling effect for the oxidative phosphorylation and the swelling of mitochondria comparing to those of general fatty acids, oleic and linoleic acids. They showed a strong lytic activity on the cell membrane as in the case of general fatty acids, linoleic, oleic, and some long chain unsaturated fatty acids. Tumor cells surviving through the treatment with unsaturated fatty acids changed the cell characteristics temporarily, with a slow-down of the ascites development and the cell growth.</p

Conformational studies of mitochondrial DNA

Ring DNA from rat liver mitochondria has been examined by circular dichroism (CD) in the region of the 225 to 320 m/~ and the followings have been clarified. The ring DNA gives a CD spectral curve somewhat different from linear DNA from nuclei, showing a big positive peak at 266 m/~ and a small negative band at 243 m!~. That is, the positive CD band of ring DNA shifted by about 7 m/~ to the shorter wavelength side from the band of the ordinary nuclear DNA, 273 m!~. Negative band appeared at the same region as that of linear DNA but reduced in depth. Heat denaturation of the ring DNA induced a red shift of the positive band, by about 4 mp., but no change in negative band. From these experimental results it has been concluded that the ring DNA has highly twisted conformation and high in G.C contents, both of which are responsible for the blue shift of the CD spectrum.</p

Observations on Nerve Cells and Fibres by Polarization Microscope

To reveal the relationship between the morphologic structure and the functioning of cells it is of great importance to know the molecular arrangement in cytoplasm. For this purpose the observation by polarization microscope is one of the most exellent methods. There are several excellent works on nerve fibre done using polarized lightl,2,3,4 but there is no report concerning the nerve cell itself. It is the purpose of this paper to show our data obtained on ganglion cells and nerve fibres with the deduced speculation on the structural arrangement of lipoprotein in the protoplasm of nerve cells and fibres.</p

Antigenicity of chlorpromazine and clozapine to rabbits

Antigenicity of clozapine was studied in rabbits, comparing with that of chlorpromazine as control. The results indicate that chlorpromazine produces antibody in rabbit as revealed by passive hemagglutination test, giving the titer of 1 : 2, 000 or higher in all the five cases observed, though specific precipitin lines has not been obtained and PCA test proves to be negative. Clozapine failed to produce anti.clozapine antibody giving negative passive hemagglutination test, passive cutaneous anaphylaxis and precipitin reaction, in all forms tested. Some remarks were made on the possible close relation between the antigenicity of the drug and its affinity to protein.</p

Detection of antibodies against histones of rat liver cells

Immunological analysis of histones extracted from the calf thymus has not been so successful because of their weak antigenicity against rabbits. Our investigations, however, have demonstrated that the histones purified from normal rat livers have the weak antigenicity against rabbits.</p

The effect of oxygen tension on tetrazolium reduction by respiratory enzyme systems of tissue culture cells

1. Attempts have been made to confirm how the formazan formation is affected in the presence of oxygen gas when the cells are incubated with neotetrazolium salt and the subsrtates for the enzymes to be tested. 2. In the cases of succinoxidase formazan formation is minimized under pure O2tension, it increases with decrease in O2 tension, and reaches its maximum value under N2gas. 3. This relationship between the oxygen tension and the diformazan formmation can likewise be observed even after pretreatment of the system with KCN. 4. In measuring enzyme activity of the DPN-diaphorase system with L-glutamate and DPN as substrate and NT as hydrogen acceptor, the same relationships between the oxygen tension and the NT-reduction can be seen as in succinoxidase system. 5. In the determination of enzyme activity of the cytochrome-c-cytochrome oxidase system with p-phenylene-diamine as substrate and NT as hydrogen acceptor, likewise the diformazan formation is markedly affected by oxygen tension and increased with the reduced oxygen tension but under pure Ns gas the value is reduced. When the systen is pretreated with KCN, however, the diformazan formation reveals its maximum value under pure nitrogen gas, the values of which correspond to those values of endogenous reaction without substrate. 6. The above results show that the neotetrazolium salt can compete with O2 as hydrogen acceptor, and less values of formazan formation may be obtained under higher oxygen tension and the higher values under lower oxygen tension independently from the true activity of the enzyme.</p

Histologic observation on the tumor tissue affected by Fatty acids from the liver of X-ray irradiated rabbits (OX)

The growth inhibitory effect of the fatty acids (OX) from the liver of X-irradiated rabbits on the solid type of Ehrlich ascites tumors has been observed both in gross and histologic observations. OX substance, a fraction of fatty acids extracted from the liver of X-irradiated rabbits has actually been found to inhibit the tumor growth by the local injection, resulting in the disappearance of the tumor after 12 injections for onemonth period, 2.4 ml of 2.5% emulsion in total dosage. Histologic observations reveal degeneration and necrosis of tumor cells, whereas in the control animals always active proliferation of tumor cells can be observed.</p