The effects of estrogen and progestin on clomiphene blocked ovulation were studied in PMS-clomiphene treated immature rats.
The animals were injected with 3 IU PMS subcutaneously at 9:3010:00 A. M. during 2426 days of age. Clomiphene (clomiphene citrate: 2.5 mg per 100 g. b. w. in 30% ethanol-saline solution) was injected subcutaneously 6 hr after PMS treatment.
Animals were killed 74 hr after PMS injection to examine the oviducts for ova, and ovarian and uterine weights were measured.
Results obtaind were as follows:
1) Ovulation was inhibited on the 3rd day morning after PMS-clomiphene treatment; ovulation delayed 24 hr by the treatment. No effect on ovulation was observed by hysterectomy (Table 1).
2) Estradiol 17-β (0.05200 μg per 100 g, b. w. in sesame oil) was singly injected at various times after PMS in PMS-clomiphene treated rats. High rate of ovulation was obtained by the treatment with 0.5 μg (4/6) and 5 μg (5/6) at 3 hrs, 5 μg (3/6) at 6 hrs, 5 μg (6/6) at 9 hrs, or 5 μg (5/6) at 51 hrs. It is suggested that estradiol can compete with clomiphene on the the receptor (s) during 3 to 9 hrs and it can stimulate release of ovulating hormone at 51 hrs after PMS injection (Table 2).
3) Progesterone (0.022 mg per 100 g b. w. in sesame oil) was injected in the same way as estradiol.
Higher ovulation rate was found in the treatment of 0.2 mg (3/4), 2 mg (3/5) at 45 hrs, 0.2 mg (2/2), 2 mg (6/6) at 48 hrs, 0.2 mg (5/6), 2 mg (5/5) at 51 hrs, and 0.2 mg (3/5), 2 mg (4/4) at 54 hrs. It is suggested that progesterone can stimulate the release of ovulating hormone during 45 to 54 hrs after PMS on the day prior to ovulation (Table 3).
4) When other steroids by single treatment with cortisone acetate, corticosterone, or 17 α-OH-progesterone was injected at 51 hrs after PMS injection in PMS-clomiphene treated rats, no ovulation occurred (Table 4).

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The effect of male proximity on the estrogen secretion was studied using the 4-day cyclic mice. The animals were housed in artificially illuminated animal rooms (light: from 5 a.m. to 7 p.m.). The female mice were divided into 2 groups. The animals of the first group were placed in cages each of which had a wire-net separated rooms for male mice. The mice of both sexes were putseparat ely in these cages from 2 p. m. on the day of vaginal estrous stage. The other groups served as a control without male mice.
1) The mice of both groups were examined for appearance of vaginal estrous stage after being ovariectomized at the several stages during diestrus and metestrus.
Appearance of the vaginal estrous stage indicated earlier secretion of ertsogen in the females placed adjacent to the male mice as compared with the control mice (Fig. 4).
2) Onset of the vaginal estrous stage was earlier in the females placed adjacent to the male mice as compared with the control mice (Fig. 2).
3) The wet weight of the uterus in the treated groups gained weight more earlier than in controls (Fig. 3).
Thus, our findings may indicate that estrogen secretion in female mice is induced by male proximity.

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The experiments were attempted to survey a possibility of fixation from carbon dioxide in the rat egg in vitro.
The eggs in 1 or 2-cell stage were collected from the animals superovulated according to the methods reported elsewhere.
For each replicate 50 to 200 eggs, depending on the stage of development, were cultured for 4 hour at 37°C in Krebs-Ringer phosphate buffer (KRP) which contained 1 μci of NaHCO₃ as basic medium with a gas phase of air. This fixation of carbon dioxide was compared to that in a various medium such as KRP+0.1% glucose+2.5 × 10^-2M of pyruvic acid (KHP+G+P), KRP+ 2.5 × 10^-1M of lactic acid (KRP+L) and KRP+10^-2M of lactic acid+ 2.5 × 10^-3M of succinic acid (KRP+L+ S) with a gas phase of air and 100% of oxygen.
After the incubation, the eggs were separated by centrifugation from the medium and recovered for further fractionation.
The eggs were recovered from the tip of the centrifuge tube and both the eggs and the incuba-tion medium were acidfied to remove unfixed carbon dioxide.
The incorporated isotopes were fractionated by the schedule in Fig. 1 and the fixed radioactivity in each fraction was estimated by liquid scintillation counter with PPO-POPOP system.
The rat eggs from 1-cell to blastocysts have fixed from carbon dioxide to organic materials in vitro.
In 2-cell stage of rat egg, the incorporation of fixed carbon from carbon dioxide was two or more times greater than that of 1-cell eggs cultured for a similar period. The labelled products in the eggs and the culture medium was greatest in the blastocysts stage.
TCA-insoluble compounds were the major part of the incorporation in the embryos. The incorporation of the eggs cultured in the medium which contained glucose and pyruvate was two times greater than that of eggs cultured in KRP and KRP+L medium. The rate of the fixed carbon had affected by the gas phase.
The metabolic pathways of the fixation of carbon dioxide in rat eggs were disscussed.

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Effects of estradiol and progesterone injection on the fructose and sorbitol formation in the uterus were investigated using seven Holstein cows (27 years of age) after ovariectomy.
Hormonal treatment was initiated on day 30 after the operation.
Injection of estradiol and progesterone, single or combined, was given intramuscularly (Exp. A) or into the uterine stroma (Exp. B), five times at 8 hr-intervals. A single dose contained 2 mg estradiol and 100200 mg progesterone in Exp. A and 2 μg estradiol and 5 mg progesterone in Exp. B.
Uterine fluid was collected for 2 hr by cannulation through the cervix, and endometrial tissue was obtained with biopsy on day 3 and on day 7 after the initial injection. Antibiotics was deposited intrauterine cavity after the sampling.
Fructose and sorbitol concentrations in the samples were estimated by gas-liquid chromatography using adonitol as an internal standard.
On day 30 after ovariectomy, fructose and sorbitol were not detectable in the uterine fluid and the endometrial tissue of the cow. Estradiol injection did not stimulate fructose and sorbitol formation. On the contrary, progeste-rone showed a stimulating effect which was further enhanced in collaboration with estrogen.
Increasing dose of progesterone to estradiol, appeared to enhance the synergistic actions of tLe two hormons.
Histologically active reactions following the ovarian hormone treatments were found in the uterine surface epithelium of ovariectomized cows.

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[in Japanese]

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The effects of eight sorts of diluent were studied on the metabolic activity of dog spermatozoa using ten adult male Spitzdogs. Spermatozoa in the second fraction of ejaculate were washed once with the diluents and resuspended in them. The results are summarized as follows:
1. In general, the metabolic activity of dog spermatozoa suspended in the diluents was higher than that in undiluted semen. The accumulation of lactic acid varied markedly according to the diluents.
2. The glycine and phosphate buffer were effective in stimulating oxygen uptake and glucose or fructose utilization of dog spermatozoa. However, the lactate was accumulated very little in these buffers.
3. Cow's milk diluent stimulated metabolic activity of dog spermatozoa markedly. Sodium glutamate diluent was shown to have a similar effect.
4. PETIC solution was effective keeping viability and glycolytic activity of dog spermatozoa.

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The effects of seven water soluble antibacterials on the viabilty and the metabolism of dog spermatozoa were compared. The antibacterials were penicillin (G cryst), kanamycin sulfate, dihydro-streptomycin sulfate, spiramycin, terramycin, sulfamine and homosulfamine. The results are summa-rized as follows:
1. The viability and the motility of spermatozoa just after the addition of antibacterials were almost the same as those in the intact semen. However, significant differences between the antibacterials were found on the survival after the incubation for two hours at 37°C. The sulfa durgs showed the best results, and spiramycia and terramycin were rather toxic on the motility of sper-matozoa.
2. In general, the metabolic activity of spermatozoa was stimulated in the addition of sulfa drugs or terramycin. Other antibacterials showed the different effects on the dog sperm metabolism, accoriding to the substrates emproyed.
3. The antibiotics except terramycin greatly reduced in lactic acid accumulation following the glycolysis or fructolysis.

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