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

Mitochondrial Swelling and Uncoupling Activity of Long-Chain Fatty Acids

The effect of various fatty acids on the swelling-contraction and oxidative phosphorylation of mitochondria from rat liver and Ehrlich ascites tumor cell have been studied and the results are as follows: 1. The swelling of rat liver mitochondria is induced by fatty acid. The extent of this uncoupling action is in the descending order of myristate, laurate, parlmitate, stearate and behenate in saturated fatty acid and linoleate, linoleneate, richinoleate and oleate in the unsaturated fatty acid. This swelling action is stronger with unsaturated fatty acids than that of saturated ones and cis form is stronger than trans form. 2. The uncoupling oxidative phosphorylation of rat liver mitochondria is also observed with these fatty acids and the activities are proportional to the degree of the swelling action. 3. The degree of swelling of rat liver mitochondria is proportional to the concentration of oleate and is inhibited by anaerobiosis and respiratory inhibitor except amytal. 4. The mitochondria swollen by fatty acid can be recontracted reversibly by ATP, Mg++ and bovine serum albumin. 5. The swelling action of sodium oleate is the strongest on mitochondria from rat liver, followed by those from the liver of Ehrlich ascites tumor bearing mouse, Ehrlich ascites tumor cells and solid Ehrlich tumor cells. 6. Sodium oleate inhibits the incorporation of 32p into ATP, ADP, GTP and UDPG in mitochondria.</p

An apparatus for simultaneous measurement of 90° light-scattering, fluorescence intensity of reduced pyridine nucleotides and oxygen consumption of mitochondria

1. An apparatus for the simultaneous measurements of volume change, fluorescence intensity of pyridine nucleotides and oxygen consumption of mitochondria has been constructed. 2. Oxygen consumption is measured by the rotating platinum electrode with a modification of Hagihara's system, attached in a cuvette of the apparatus. 3. Volume changes of mitochondria (swelling-shrinkage) are measured by the 90° light-scattering at 650 m&#956;. 4. Relative fluorescence intensity of pyridine nucleotides is measured by the fluorometer: for the excitation, a bright light at 365 m&#956;. line of mercury lamp is isolated through the filter and exposed to the mitochondria suspended in a cuvette of the apparatus, and fluorescent emission is analyzed by a grating mirror monochromator. 5. The scattered light at 650 m&#956;. is not affected by the excitation light and the fluorescent emission, and fluorescence intensity is not affected by the scattered light at 650 m&#956;. 6. The simultaneous measurements of the oxidation-reduction of pyridine nucleotides, the respiration states and the changes in the intensity of 90° lightscattering of mitochondria are given as an example of the performance of the present apparatus.</p

The effects of high fatty acid on the glucose metabolism of Ehrlich ascites tumor cells

The effects of high fatty acids such as oleic, richinoleic, linoleic, linolenic, palmitic and stearic acids, on the respiration, glycolysis, organic phosphate synthesis of Ehrlich ascites tumor cells, were studied. The unsaturated fatty acids added to the media enhanced the respiration of the tumor cells at the concentration lower than 0.2 mM, after a short incubation period and inhibited the respiration in a high concentration 0.4 mM. The saturated fatty acids did not show such effect. All the fatty acids, both of saturated and unsaturated, effected the increase in lactate formation in tumor cells, especially markedly at higher concentration being accompanied by the WQ increase and RQ around 1. The respiration lowered by the fatty acids was ameliorated by the addition of glucose. The lactate formation from glucose was greatly enhanced by the addition of fatty acids but hardly from pyruvate. The unsaturated high fatty acids proved to have a strong uncoupling action for oxidative phosphorylation. This effect could be recognized slightly in the saturated fatty acids. The addition of high fatty acid resulted in the striking decrease in ATP and ADP with the increase in AMP. With these results the discussion was conducted concerning the specificity of tumor cell related to the glucose and fatty acid metabolism.</p

Studies on swelling-shrinkage and oxidative phosphorylation of liver mitochondria of rat fed on 3'-methyl-4-dimethylaminoazobenzene

The swelling-shrinkage and oxidative phosphorylation of rat liver mitochondria affected by 3'-methy1-DAB feeding were observed in correlation with function by the method mentioned, and the following results were obtained: 1. By feeding 3'-methy1-DAB the swelling-shrinkage ability of rat liver mitochondria showed a remarkable alteration reducing in the amplitude. It reduced gradually during the days of feeding, reached the minimum value on 30th day and restored gradually thereafter (in Case 1). 2. ADP/O ratio also decreased by feeding the carcinogen reached the minimum point on 30th day and increased on 38th day showing the similar tendency in the decrease of the swelling-shrinkage amplitude (in Case 1). 3. The mitochondria from the hepatoma, which was induced by 3'-methy1DAB feeding, showed a lower amplitude in swelling-shrinkage with the dropped ADP/O ratio compared with those of mitochondria from liver tissue neighbouring the tumor. 4. The mechanism in the reduction of swelling-shrinkage ability has been discussed in the relation with fatty acid composition of mitochondria which is reported elsewhere. 5. From the above results it is deduced that lowered ability for swellingshrinkage with the reduced oxidative phosphorylation will be somehow related to the mechanism of cancer induction.</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