[Effects of night shift on plasma concentrations of melatonin, LH, FSH and prolactin, and menstrual irregularity].

To examine the effect of night shift on the ovarian function, 122 teachers, 67 office workers, 377 nurses, 133 factory workers and 67 barmaids were surveyed. The incidence of irregular menstrual cycle was 13.1% in teachers, 14.9% in office workers, 24.9% in nurses, 36.8% in factory workers and 40.3% in barmaids. The incidence was significantly higher in women working at night than women working during the day. Plasma concentrations of melatonin, LH, FSH and prolactin were determined at 2200 h and 0200 h in 5 nurses working at night and in 6 nurses resting in their quarters. Plasma concentrations of melatonin and prolactin at 0200 h were significantly lower in nurses of the working group than others of the resting group, but plasma concentrations of LH and FSH did not differ between the two groups. These results indicate that night shift suppresses the ovarian function by affecting the circadian rhythm of melatonin and prolactin.

[Effects of continuous lighting on secretion of melatonin and pituitary hormones in women].

Effects of light exposure on the serum concentrations of melatonin, prolactin, LH and FSH were studied in 53 women during their follicular phases. Twenty seven women were exposed to light (500-800 lux at eye level) from 17:30h to 22:00h, and 17 women received the same light intensity from 17:30h to 2:00h, while 9 women stayed in a dark room from 21:00h to 2:00h as control subjects. The light exposure suppressed a nocturnal increase in the serum concentrations of melatonin both at 22:00h and 2:00h. The light exposure and awake condition suppressed the serum prolactin concentrations and increased the serum FSH concentrations at 2:00h, while the influence on the serum LH concentrations was not clear because of LH pulsatility. These results show that the circadian rhythm of melatonin, prolactin and FSH is affected by light exposure and wakefulness, and the effects of light exposure would become more evident if light exposure was continued over a longer period.

Morphologically and functionally distinct subpopulations of steroidogenic cells in corpora lutea during pregnancy in rats.

On days 5, 10, 15 and 20 of pregnancy, rat corpora lutea (CL) were dissected and dissociated into single cell suspensions by enzyme treatments. The suspended luteal cells were allowed to sediment in a BSA gradient at 4 degrees C for 3.5 hours. Five fractions were collected from the top (Fraction (Fr.) 1) to the bottom (Fr. 5) of the gradient. Cells were incubated in serum-free DME-F12 for 20 hours with or without hCG (100 ng/ml) to test them functionally, and the accumulation of progesterone and testosterone was determined by radioimmunoassay. To assess 3,3-hydroxysteroid dehydrogenase (HSD) activity, a histochemical suspension-staining procedure was used. Cells were examined by light microscopy, and the percentage of cells containing dark blue formazan deposits and their diameters were determined in at least 40 microscopic fields. The number of cells staining for 3 beta-HSD did not vary by day 15 but decreased from 141.6 +/- 16.5 X 10(3) cells/CL on day 15 to 113.8 +/- 13.2 X 10(3) cells/CL on day 20 of pregnancy. However, 3 beta-HSD-positive cells maintained the same levels of progesterone secretion until the advent of luteolysis, then they increased in size progressively throughout pregnancy. In BSA gradients, the relatively larger 3 beta-HSD-positive cells migrated faster than the smaller 3 beta-HSD-positive cells on each day of pregnancy. The diameters of 3 beta-HSD-positive cells differed significantly in Frs. 2, 3, 4 and 5 on days 15 and 20 of pregnancy. On day 15 of pregnancy, less progesterone accumulated in wells containing 3 beta-HSD-positive cells from Fr. 2 (mean diameter; 24.96 microns) than from Fr. 3, Fr. 4 and Fr. 5 (mean diameters; 27.20, 30.79 and 31.28 microns, respectively) but the Fr. 2 cells responded more to hCG stimulation. Fr. 2 also showed a higher ratio of testosterone accumulation to progesterone accumulation than the other fractions. The response to hCG stimulation of cells in Fr. 2 tended to be higher than that in Fr. 3 on day 20 of pregnancy. These data suggest that the steroidogenic rat luteal cells are comprised of morphologically and functionally different cell types after day 15 of pregnancy. No stimulating nor inhibiting effects were observed in co-incubation of cells from Fr. 2 with cells from Fr. 3 or Fr. 4 on days 15 and 20 of pregnancy.

[Clinical significance of measurement of plasma melatonin concentration in women with irregular menstrual cycles].

To study the effects of melatonin on the human hypophyseal ovarian axis, we measured the concentrations of melatonin in the plasma of women with a regular menstrual cycle and of women with an abnormal menstrual cycle, at days 5 through 10 after the onset of menstruation or withdrawal bleeding. In seven women with a regular menstrual cycle, there was a tendency for the plasma melatonin concentration to gradually decrease chronologically from 49.0 +/- 9.8pg/ml (M +/- S.E.) at 9:30 h to 19.0 +/- 4.0pg/ml at 12:00 h. On the other hand, the plasma melatonin concentration at 21:30 h was 50.3 +/- 7.4pg/ml which was almost the same as the level at 9:30 h, while the concentration during the night significantly increased chronologically to 106.3 +/- 28.6pg/ml at 24:00 h. By comparing the plasma melatonin concentration measured during the day with that measured during the night, a significant difference was discovered after 21:40 h (p less than 0.05). The plasma melatonin concentration at 10:00 h and 22:00 h in 20 women with a regular menstrual cycle was 28.1 +/- 3.4pg/ml and 72.2 +/- 9.9pg/ml, respectively, showing a diurnal rhythm with a significantly higher level during the night than during the day. On the other hand, the plasma melatonin concentration in most of the women with an anovulatory cycle or amenorrhea was within the range of the mean level of +/- 1.5 x the standard deviation obtained from the 20 women with a regular menstrual cycle. However, an increase in plasma melatonin during the night was not found in three of nine women with an anovulatory cycle and one of two women with second grade amenorrhea.(ABSTRACT TRUNCATED AT 250 WORDS)

[The effects of light exposure on plasma concentrations of melatonin, LH, FSH and prolactin in women].

The effects of light exposure on plasma concentrations of melatonin, LH, FSH and prolactin were studied in 11 normal cycling women during their follicular phases. Blood samples were obtained via an indwelling venous catheter every 10 min. for 2.5 hours starting at 9:30 and 21:30h. For the blood samplings taken at night, six women were kept in a dark room and were permitted to sleep. Their blood samples were obtained using a flashlight (5-10 lux) without their rest being disturbed. However, the other five women were exposed to light (3,000 lux at eye level) and awakened from 22:40 to 24:00h. Plasma melatonin concentrations in the morning decreased from 48.7 +/- 11.6 pg/ml at 9:30h to 24.7 +/- 4.0 pg/ml at 12:00h. On the other hand, plasma melatonin concentrations at night increased from 65.4 +/- 9.6 pg/ml at 21:30h to 138.2 +/- 28.6 pg/ml at 24:00h. The pulsatile LH secretion was changed from the type of "high frequency, low amplitude" in the morning to the type of "low frequency, high amplitude" at night. Nocturnal FSH concentrations were lower than diurnal ones, but nocturnal prolactin concentrations were higher than diurnal ones. Nocturnal concentrations of melatonin were suppressed 40 min. after the light exposure (from 117.4 +/- 11.4 pg/ml at 22:40h to 74.6 +/- 13.9 pg/ml at 23:20h). On the the other hand, the light exposure increased plasma prolactin concentrations from 10.9 +/- 4.1 ng/ml at 22:40h to 17.0 +/- 4.4 ng/ml at 22:50h, maintained those higher levels for 20 min. and decreased them gradually after 23:20h. With the light exposure, mean values of nocturnal LH concentrations were increased from 11.9 +/- 1.5 mIU/ml before exposure to 14.2 +/- 1.8 mIU/ml after exposure, and those of FSH were also increased from 5.9 +/- 0.4 mIU/ml to 6.3 +/- 0.4 mIU/ml. These results showed that the secretion of melatonin, as well as LH, FSH and prolactin had daily rhythms and that melatonin and prolactin showed different responses to light exposure, suggesting different control mechanisms for the secretion of those two hormones.

[Morphological and functional changes in 3 beta-HSD positive cells in corpora lutea during pregnancy in rats].

On days 5, 10, 15 and 20 of pregnancy, rat corpora lutea (CLs) were dissected and dissociated into single cell suspensions by enzyme treatments. To assess 3 beta-hydroxysteroid dehydrogenase (HSD) activity, a histochemical suspension-staining procedure was used. The number of cells positive for 3 beta-HSD in CL were 148.0 +/- 13.7, 130.1 +/- 25.4, 134.0 +/- 23.5 and 116.8 +/- 13.5 X 10(3) cells on days 5, 10, 15 and 20 of pregnancy, respectively. The 3 beta-HSD positive cells increased in size from 18.5 +/- 0.28 microns on day 5 to 35.7 +/- 0.50 microns on day 20 of pregnancy. The suspended luteal cells were incubated in serum-free DME-F12 for 20 hours with or without human chorionic gonadotropin (hCG, 100 ng/ml) or dibutyryl cyclic AMP (dbcAMP, 10mM) to test their functionality, and progesterone accumulation was determined by radioimmunoassay. Progesterone secretion from the 3 beta-HSD positive cells was maintained at the same levels until day 15 and decreased significantly on day 20 of pregnancy. However the response to hCG stimulation of the 3 beta-HSD positive cells decreased significantly on day 20, the 3 beta-HSD positive cells maintained the same responsiveness to dbcAMP stimulation on progesterone secretion throughout pregnancy. These data suggest that the steroidogenic rat luteal cells may be regulated by morphologically and functionally different mechanisms, and that progesterone may be secreted by at least two different pathways.

[Regulation of progesterone accumulation in dissociated luteal cells on day 15 of pregnancy in rats].

Luteal cells from the corpora lutea of rats on day 15 of pregnancy were suspended in DMEM-F12 and progesterone accumulation was measured by radioimmunoassay. Progesterone accumulation increased steadily over eight hours and then gradually decreased. Progesterone accumulation was seen to increase with the addition of BSA and 25-hydroxycholesterol. At doses of 12 ng/ml and 600 ng/ml hCG, progesterone accumulation increased, but was seen to decrease at a dose of 30 micrograms/ml hCG. However, the addition of cyclohexamide alone had no effect on progesterone accumulation, but the simultaneous addition of cyclohexamide and hCG blocked the effects of hCG in a dose related manner. These data indicate that progesterone production by rat luteal cells depends on cholesterol availability. The results reveal that hCG stimulates progesterone production by a process requiring new protein synthesis.

[Changes in ovarian 3 beta-hydroxysteroid dehydrogenase and 20 alpha-hydroxysteroid dehydrogenase activities in pregnant rats].

Changes in ovarian 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) and 20 alpha-HSD activities were studied in pregnant rats. The activity of 3 beta-HSD was determined by measuring the rate of conversion of pregnenolone to progesterone. The activity of 20 alpha-HSD was determined by measuring the rate of conversion of 20 alpha-hydroxy-pregn-4-en-3-one to progesterone. 3 beta-HSD activity in corpora lutea (CL) was low between days 8 and 12, increased rapidly on day 15, and then gradually decreased until day 23. 20 alpha-HSD activity in CL markedly decreased from days 8 to 18, and then increased until day 23. The activities of 3 beta-HSD and 20 alpha-HSD in non-luteal ovarian compartment (NLO) showed no significant changes between days 8 and 23. These results indicated the important role of these enzymes in the secretion of progesterone from CL. It was also suggested that these enzyme activities in NLO might be controlled by a different mechanism.

A quantitative relationship between conceptus number and ovarian steroid dehydrogenase activity in pregnant rats.

The effects of a conceptus-derived substance on the activity of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) and 20 alpha-HSD in the ovary were studied in the rat. On day 7 of pregnancy (day 1 = insemination), rats were laparotomized and the desired number of conceptuses was aspirated from the uterus; thus, rats carrying one, two, three, four, five to seven or eight to ten conceptuses were prepared. They were autopsied on day 15 and 3 beta-HSD and 20 alpha-HSD activity in the corpus luteum (CL) or non-luteal ovarian tissue ( NLO ) was determined. Conceptus number was directly related to 3 beta-HSD and inversely related to 20 alpha-HSD activity in the CL. The serum progesterone level and CL weight were also directly related to conceptus number. Neither 3 beta-HSD nor 20 alpha-HSD activity in the NLO was affected by conceptus number. These results indicated that 3 beta-HSD and 20 alpha-HSD in the CL are probably regulated by placental hormone secreted in proportion to the number of conceptuses; in the NLO these enzymes may be controlled by a different mechanism.

Role of the nongravid part of the uterus in the regulation of corpus luteum function in pregnant rats.

The effect of the nongravid part of the uterus on corpus luteum (CL) activity was studied in rats bearing a single conceptus during midpregnancy. On day 7 (day 1 = insemination), the number of conceptuses was reduced to one by aspirating all but one conceptus (the aspiration group), by removing all of of the uterus except that containing a single conceptus (the hysterectomy group), or by aspirating conceptuses, as in the aspiration group, plus ligating the uterus, as in the hysterectomy group, but without removing the uterus (the sham hysterectomy group). Rats with a full complement of conceptuses (the intact control group) served as additional controls. Blood samples were obtained on days, 10, 12, and 15, to determine the effects of these procedures on the increase in serum progesterone that normally occurs between days 12 and 15 and to observe any related changes in serum LH between days 10 and 15. On day 15, the mean serum prgesterone concentrations and weights of the CL in the aspiration and sham hysterectomy groups were significantly lower than those in the intact control group. In the hysterectomy group, however, serum progesterone was even higher than in the intact control group, although the CL were significantly smaller than in the intact control group and were not different from those in the aspiration or sham hysterectomy groups. Serum LH on day 10 was significantly lower in the hysterectomy group than in any of the other groups; the latter were not significantly different from each other. By day 15, serum LH in the intact group had fallen to less than 50% of the value on day 10, while in the aspiration group, it remained as high as on day 10. In the hysterectomy group, there was no significant difference in serum LH concentrations between days 10 and 15. These results suggest that the nongravid part of the uterus inhibits progesterone secretion, and that in its absence, even a single conceptus can increase progesterone secretion after day 12 as much as can a full complement of conceptuses, but without also increasing the size of the CL. The uterus may exert this effect either through the pituitary or in cooperation with a pituitary factor, possibly LH.

[The double contrast hysterography using CO2 insufflation (author's transl)].

The double contrast hysterography, which was performed by the continuous CO2 insufflation with the ordinary HSG, was applied to 162 infertile patients. Forty-nine of 162 patients showed the abnormal findings by this method. Hysteroscopic or histological examinations revealed that 26 of these patients had hyperplasia, 15 had polyp, 2 had myoma and 1 had intrauterine foreign body. When these results were compared with those obtained by ordinary HSG, it was found that the double contrast hysterography would be more useful to investigate the detailed intrauterine changes such as hyperplasia or polyp than the ordinary HSG, and, therefore, would be a useful aid in the clinical care.

Effects of domperidone on serum prolactin levels in human beings.

Effects of domperidone, a dopaminergic antagonist, on serum prolactin levels were studied in 6 normal men and 6 normal cyclic women at the different phases of their menstrual cycles (i.e., the follicular, the preovulatory and the luteal phases). Domperidone (10 mg, i.v.) caused significant increases in serum prolactin in all cases within 15 min after the injection. The prolactin response was significantly (p < 0.01) higher in women than in men, and there was no significant difference in the prolactin responses among the three phases of the menstrual cycles. These results indicate that domperidone may be an effective stimulator of serum prolactin secretion in human beings.

Tumor antigen of human cervical squamous cell carcinoma: correlation of circulating levels with disease progress.

A double-antibody radioimmunoassay method was used for serial determinations of a tumor-antigen (TA-4) of cervical squamous cell carcinoma, and the correlation of serum antigen levels with the disease progess was investigated in 23 patients with cervical squamous cell carcinoma. Ten cases with widespread metastases received radiotherapy and/or chemotherapy. Nine of these cases who showed progression of the disease had a corresponding increase in serum antigen levels, while one case who had regression of the disease showed a corresponding decrease in serum antigen levels. Thirteen patients received radical surgery, and in all of these, high pretreatment antigen levels declined to undetectable levels 1 or 2 weeks after surgery. A panel of coded sera from the NCI-Mayo Clinic Serum Bank was also studied for evaluating the specificity of the assay. Thirteen of 25 patients (52%) with cervical squamous cell carcinoma showed positive serum antigen levels, while only one of 58 control cases (1.7%) showed false-positive result. These results suggest that serial TA-4 determinations may provide a useful method for evaluating regression or progression of the disease.