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

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

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

Uptake of nicotinamide adenine dinucleotide and excretion of its degradation pro­ducts by tissue culture cells

For the purpose to reveal the mechanism of uptake and degradation of NAD by cells, the authors conducted the observation on the L cells cultured in the medium containing NAD and the following results have been obtained. 1. NAD in the medium is taken up by the cells in its intact form, reaching about twice the value of the control. 2. The spontaneously degraded products of NAD, nicotinamide and adenine dinucleotide ribose, in the same molar concentration as NAD used in the present experiment, have no effect on the NAD content of L cells. 3. The NAD taken up by the cells is degraded into nicotinamide mononucleotide (NMN) and adenine mononucleotide (AMP) by pyrophosphatase including NADpptase and excreted in the medium. Unexpectedly the ingested NAD is not degraded by NADase in the L cell. 4. L cells metabolize the same amount of NAD as that contained originally in the cell for about ten minutes, as calculated from the amount of NMN excreted in the medium.</p

A new method for counting the reticulocyte number

The counting of reticulocyte number by the routine method on the dye fillms often leads to a poor result. This can be avoided by counting them on the collodion dye film on which the almost equal distribution of reticulocytes can be attained.</p

Cellular response to the ribonuclease injection; a morphologic and cytochemical study

By the repeated injection of RNase into mice the histological, cytochemical and electronmicroscope observations of several tissues and the quantitative estimation of DNA contents per cell in liver have been conducted. The observations proved that the most marked changes occur in basophilia, ER (endoplasmic reticulum) and Palade's granules; the dissociation of the granules from ER and their agglomeration, and the final disappearance of the granules and ER. The increase of the granules in number surrounding the nucleus seen in liver cells and the appearance of the ring form ER in the pancreatic exocrine cells and its development from the nuclear membrane have been traced morphologically and these are comprehended as the regenerating picture of ER and granules from the nuclear outer membrane. DNA contents in liver cell increase in the early stage and decrease to the normal level in the later stage. The former is attributed to the cessation of mitosis by the damage of cell center without interference on DNA synthesis and the latter to the disappearance of the cells of tetraproidy by degeneration.</p