Effects of some diethylstilbestrol metabolites and analogs on cytotoxicity and aneuploidy induction in Chinese hamster V79 cells.

We have previously reported the inhibitory effects of diethylstilbestrol (1) and optically active indenestrol derivatives on microtubule polymerization in vitro and their disruptive effect on cytoplasmic microtubules and cytotoxicity in cultured Chinese hamster V79 cells. In the present study, the cytotoxicities of (+-)-diethylstilbestrol oxide (2), (+)-, (-)- and (+-)-monomethyl ethers (4) of 2, (+-)-dimethyl ether (5) of 2, diethylstilbestrol pinacolone (3), E,E-dienestrol (6), Z,Z-dienestrol (7), meso-hexestrol (8), a mixture of (1R,1'S)4-hydroxyhexestrol and (1R,1'S)4'-hydroxyhexestrol (9), and the 4-hydroxy derivative (10) of diethylstilbestrol dimethyl ether were investigated in Chinese hamster V79 cells. The results indicated that the cytotoxic activity of 10 was the strongest of the compounds tested, although its activity was the almost same as that of 1. Moreover, as the activity of (-)-4 was greater than those of 2 and 1 monomethyl ethers, the effect of 4 on cytotoxic activities was elucidated. In conclusion, the present results indicate that the cytotoxic activities of hydroxylated metabolites are greater than those of each mother compound, although epoxidation of 1 leads to a product which can be broken down more readily than the parent compound.

Characterization of semiquinone free radicals formed from stilbene catechol estrogens. An ESR spin stabilization and spin trapping study.

Electron spin resonance spectroscopy has been used to detect, characterize, and to infer structures of o-semiquinones derived from stilbene catechol estrogens. Radicals were generated enzymatically using tyrosinase and were detected as their Mg2+ complexes. It is suggested that initial hydroxylation of stilbene estrogen gives a catechol estrogen in situ; subsequent two-electron oxidation of the catechol to the quinone, followed by reverse disproportionation, leads to the formation of radicals. Consistent with this mechanism, o-phenylenediamine, a quinone trapping agent, inhibits formation of o-semiquinones. A competing mechanism of radical production involves autoxidation of the catechol. Hydroxyl radicals are shown to be produced in this system via a mechanism involving reduction of iron and copper complexes by stilbene catechols. Possible differences in the reactivity of stilbene ortho- and para-semiquinones are discussed.

Performance of Leghorn type hens fed two levels of energy and a synthetic estrogen during the growing period.

Feeding of an estrogen, dienestrol diacetate, at 352 mg/kg of diet to December-hatched White Leghorn type pullets, from 16 to 20 wk of age, caused onset of production to be delayed approximately 3 wk. The dietary dienestrol diacetate also resulted in increased body weights at 30 and 46 wk in one experiment. Hens receiving the estrogen laid significantly (P less than .05) heavier but fewer eggs during most of the production year than did those fed diets without the estrogenic compound. The addition of 2.2% fat to diets of pullets from 0 to 20 wk of age failed to influence their performance in the layer house.

Reduction in hepatic lipid and plasma estradiol in estrogenized chicks injected with ascorbic acid.

Injecting White Leghorn chicks every other day with 20 mg ascorbic acid significantly reduced the increase in liver weight and lipids caused by feeding a diet with 0.1% dienestrol diacetate. In chicks fed two different basal diets containing 0.1% dienestrol diacetate, injecting chicks every other day with 20 mg alpha-tocopherol did not significantly reduce liver weight or lipids while the ascorbic acid injections did. Injecting meat-type chicks implanted with estradiol with 10 mg ascorbic acid daily significantly reduced liver weight, liver lipids, and plasma estradiol, but injecting with 8 mg alpha-tocopherol daily had no significant effect.

Effect of diisopropyl 1,3-dithiolan-2-ylidenemalonate on experimental fatty livers in chicks.

The effect of diisopropyl 1,3-dithiolan-2-ylidenemalonate (NKK-100) on experimental fatty livers was investigated in chicks administered an antithyroid agent plus synthetic estrogen or in estrogenized, starved-refed chicks. NKK-100 was added at levels of 250, 500, 1000, and 1500 mg/kg diet. Liver weight was significantly decreased by administration of NKK-100 at 1500 mg/kg diet in the estrogen-administered chicks. Liver lipid content and liver lipid deposition were significantly and inversely decreased with increasing NKK-100 concentration in the diet in the estrogen administered chicks. Plasma transaminase activity, which was elevated by the estrogen administration, was reduced by the administration of NKK-100. These results suggest that NKK-100 may be of value in preventing fatty livers in poultry.

The influence of environmental temperature, thyroid status and a synthetic oestrogen on the induction of fatty livers in chicks.

1. Hepatic lipid content, lipogenic enzyme activity and plasma lipid concentration were measured in chicks reared at 21 degrees or 34 degrees C and after thyroxine (T4), thiouracil (TU), propylthiouracil (PTU), dienestrol diacetate (DD) or PTU with DD had been given for 14 d. 2. At 34 degrees C there was a significant increase in the total liver lipid and triglyceride content. 3. Injections of T4 decreased liver lipid content whereas it was increased by feeding PTU or DD. The effects of PTU were more pronounced at 21 degrees C while those of DD were more pronounced at 34 degrees C. 4. There were significant interactions between temperature, thyroid status and synthetic oestrogen treatments on total lipid and triglyceride content of the liver. Fatty liver with marked steatosis could be produced through synergic actions of PTU and DD in chicks maintained at 21 degrees C.

Hormonal regulation of a new female-specific serum protein (FP) of the laboratory rat.

This paper reports evidence for a hormonal regulation of a new rat serum protein (female specific protein, FP) which is only demonstrable in female rats. Male and female rats were treated with testosterone and estrogen. The FP was assayed with immunological methods. The following results were obtained: 1. In testosterone-treated females the serum level of FP is reduced significantly 2. In estrogenized males the FP was distinctly demonstrable but not in the control males.

Synthesis of metabolic intermediates of diethylstilbestrol.

This report details the synthesis of 1) 3,4,4'-trihydroxy-alpha, alpha'-diethyl-trans-stilbene; 2) 3,4-bis-(p-hydroxyphenyl)-trans-3-hexenol; 3) 3,4-bis-(p-hydroxyphenyl)-2,4-cis,cis-hexadienol; 4) 3,4-bis-(3'-methoxy-4'-hydrophenyl)-trans-3-hexene; 5) 3,4-bis-(3',4'-dimethoxyphenyl)-trans-3-hexene. These compounds are suspected metabolites of diethylstilbestrol.