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SUMMARY

Intact female BALB/cCrgl mice, 3–4 weeks old, were pretreated with oestrogen and progesterone for 9 days. Whole mammary glands from these mice were cultivated for 5 days in a synthetic medium supplemented with aldosterone (A), prolactin (MH) and insulin (I), with and without thyroxine (T₄) at concentrations ranging from 0·01 to 5 μg./ml.

A medium containing 1 μg. A +5 μg.MH +5 μg.I/ml. was generally optimal for lobulo-alveolar development. Addition of thyroxine to this combination resulted in a decrease in development which was highly significant at higher concentrations. However, when cultures were maintained in media containing suboptimal or low amounts of prolactin (1 μg. A + 3 μg. MH +5 μg. I/ml. and 1 μg. A + 1 μg. MH +5 μg. I/ml., respectively), the results indicate two possible effects of thyroxine: lower amounts of thyroxine had synergistic effects, whereas greater amounts had antagonistic effects on lobulo-alveolar development.

Endocrine Research Laboratory, Department of Obstetrics and Gynecology, Howard University Medical School, Washington D.C. 20059, U.S.A.

(Received 13 March 1978)

Cicero, Bell, Wiest, Allison, Polakoski & Robins (1975) and Mendelson, Mendelson & Patch (1975) reported that treatment of men with methadone decreases the concentration of testosterone in the blood, whereas Cushman (1973) found no significant decrease in his study. The direct effect of methadone on the capacity of the human testes to synthesize [³H]testosterone from [³H]cholesterol has therefore been investigated.

Human testicular tissue (0·5 g, obtained fresh at autopsy) was chopped with scissors and incubated with purified [1,2-³H]cholesterol (5·0 μCi, sp. act. 60 Ci/mmol; New England Nuclear Corporation, Boston, Massachusetts) in 5·0 ml Earle's medium (Difco, Detroit, Michigan) for 3 h with air as the gas phase. Co-factors were added as described by Ahluwalia, Williams & Verma (1974). Methadone hydrochloride (6-dimethylamino-4,4-diphenyl-3-heptanone hydrochloride; Eli-Lilly Co., Indianapolis, Indiana) or human chorionic gonadotrophin

In a previous study, it was shown that thyroid hormone secretion rate (TSR) of female rats aged 55 days was reduced 22·8% by the s.c. injection of 50 μg. melatonin/day, by 14·8% when 75 μg./day was injected at 85 days, and by 9·1% when 100 μg./day was administered at 115 days (Narang, Singh & Turner, 1967). Since the rats were increasing in body weight during the period, the decrease in TSR with age may have been due to the effects of a lower dose of melatonin/g. body weight (Narang & Turner, 1966; Kumaresan & Turner, 1967). The object of the present experiment was to determine the effect of graded levels of melatonin upon the TSR of groups of rats of the same age and body weight and to determine whether melatonin has an effect on TSR of mature animals.

The TSR of 62 Sprague—Dawley—Rolfsmeyer female rats were estimated at 33

SUMMARY

One hundred and twenty albino female rats (Sprague-Dawley-Rolfsmeyer) were divided into five equal groups. Rates of thyroxine secretion (TSR) and food consumption were determined during the control period, and 10 and 25 days after initiation of dietary treatment. Animals in each group served as their own controls for the following modifications of their diet: (1) protein-free diet, (2) 5% protein (casein) diet, (3) 10% protein diet, (4) 15% protein diet, and (5) 20% protein diet. Purina lab chow (23·4% protein) and the 20% casein diet served as control diets. The TSR, the body weight and amount of food consumed were depressed significantly in the group fed on a protein-free diet for 10 and 25 days. The group fed 5% protein diet had a non-significant decrease in TSR as compared with the controls. Similarly, TSR was not reduced by 10, 15 or 20% protein diets. Food consumption decreased significantly in the groups fed a 5, 10 and 15% protein diet, but not in the group on 20% casein. Body weight decreased significantly in the groups on a protein-free diet and on a 5% protein diet.

It would appear from these results that protein content of the diet does not become a limiting factor for TSR until it is lower than 5%. It is suggested that the calorie intake plays a more important role in regard to TSR than a low protein content of the food.

ABSTRACT

Dietary factors are known to modulate concentrations of sex hormone-binding globulin (SHBG). In the present study we have investigated the possibility that insulin like growth factor-type I (IGF-I) may be an additional regulator of SHBG using cultured human hepatoma cells which secrete SHBG. The inhibitory effect of insulin on SHBG secretion by these cells was confirmed but, in addition, IGF-I was shown to inhibit SHBG secretion by about 40% at a concentration of 100 nmol/l. A similar degree of inhibition was achieved using insulin at a concentration of 10 umol/l. Insulin, but not IGF-I, was also found to inhibit the secretion of a low molecular weight IGF-binding protein (IBP-I), which is also secreted by hepatoma cells. It is concluded that IGF-I is an additional regulator of SHBG secretion by these cells and that it may be involved in regulating SHBG secretion in vivo in response to dietary factors.