• 한국가축번식학회지
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• 제14권4호
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• pp.281-287
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• 1990

This study was conducted to find out the changes of ovarian, placental and fetal weights and periods of pregnancy in rats implanted with progesterone-tube during the reproductive stages. One hundred and thirty-four mature rats, 10~13 weeks old, were offered for this experiment. The animals, which were implanted with silicon tubes filled with progesterone on day 15 of pregnancy, were sacrified at 18, 20, 21 and 22 days of pregnancy. The changes of ovarian, placental and fetal weights and the number of fetuses during late pregnancy were recorded. The results obtained were summarized as follows : 1. After progesterone-tube implantation, ovarian weight reached to a peak value of 92.0$\pm$0.9mg at 20 days of pregnancy, there after decreased significantly to 79.5$\pm$7.6 and 68.26$\pm$4.2mg at 20 and 22 days of pregnancy(P<0.01). 2. The placental weight increased rapidly during 15~18 days of pregnancy in control and progesterone treated rats. A peak value of 447.78$\pm$20.9mg was shown at 20 days of pregnancy after progesterone-tube implantation, and in control rats the value decreased significantly to 419.42$\pm$11.6 and 404.1$\pm$29.3mg at 20 and 21 days of pregnancy(P<0.01). 3. The fetal weights was not shown any significant differences between control and progesterone-tube implanted rats. 4. The number of fetuses in control rats were 14.75$\pm$0.4 at 8~10 days of pregnancy and 13.5$\pm$0.3 and 13.25$\pm$0.4 at 12 and 20 days of pregnancy. 5. The significant difference in periods of pregnancy was appeared between progesterone-tube implanted(27.3$\pm$0.3 days) and control(22.1$\pm$0.3 days)rats(P<0.01).

• 한국가축번식학회지
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• 제14권4호
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• pp.273-280
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• 1990

This study was conducted to find out the changes of progesterone and 20$\alpha$-dihydroprogesterone(OHP) levels in the serum of female rats by progesterone-tube implantation during late pregnancy. One hundred and twenty rats, 10-13 weeks old, were offered for this experiment. Blood samples were taken from the rats on 18, 20, 21 and 22days of pregnancy. The rats were implanted with silicon tubes filled with pregesterone on day 15 of pregnancy. The progesterone and 20$\alpha$-OHP in serum were assayed by radioimmunoassay. The results obtained are summarized as follows : 1. The progesterone levels in rats of control group showed 126.36$\pm$20.19ng/ml on 18days and gradually decreased to 69.3$\pm$11.9, 29.2$\pm$1.8ng/ml 20 and 21 days. 2. The progesterone levels after progesterone-tube implantation showed 120.1$\pm$8.5, 59.01$\pm$3.1, 85.33$\pm$17.9 and 62.9$\pm$6.1ng/ml on 18, 20, 21 and 22days of pregnancy, respectively. 3. The 20$\alpha$-OHP levels in rats of control group showed 68.5$\pm$8.0ng/ml on 18 days and gradually increased to 139.9$\pm$3.7 and 141.4$\pm$6.7ng/ml on 21 and 22days. 4. The 20$\alpha$-OHP levles after progesterone-tube implantation showed 108.5$\pm$32.7 and 106.4$\pm$10.6ng/ml on 18 and 20days and increased to reach the peak level at 21days(225.01$\pm$9.9ng/ml) and rapidly decreased on 22days(85.9$\pm$10.7ng/ml).

• 한국가축번식학회지
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• 제9권2호
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• pp.105-112
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• 1985

This research was carried out to investigate the optimal conditions ssociated with the RIA procedures such as a bridging phenomena, prozone effects and a new separation methods etc. The results obtained were summarized as follows: 1. The lgG fractions of donkey anti-rabbit IgG sera were purified by Protein-A-Sepharose affinity column, which indicates that Protein-A an affinity for IgG class of donkey antiserum. 2. In coating the IgG fraction on polystryene tubes, incubation conditions made no differences between 2 hr at room temperature and overnight at 4$^{\circ}C$. 3. There were no significant differences between 1st antibody-coated tube and 2nd antibody-coated tube as a separation method when compared in terms of reproducbility. A better reproducibility may be expected if the titers of 1st antibody for the progesterone to be assayed and of corresponding 2nd antibody are reasonably high. 4. The titers of anti-progesterone antibody for 3H-progesterone and progesterone-11HS-125I were 1:300 and 1:700 in liquid-phase, and 1:100 and 1:300 in solid-phse for the separation methods. 5. A bridging phenomena in which a standard curve is long and shallow were observed when progesterone-11HS-125I was used for the tracer, but not in 3H-progesterone. 6. A prozone effect in a solid-phase system, especially 1st antibody-coated tube method was observed which the degree of inhibition was significantly different although zero bindings look the same. In this case, the titration curve should be made both in the absence and in the presence of a, pp.opriate amount of competiter, standard, respectively.

• Clinical and Experimental Reproductive Medicine
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• 제1권1호
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• pp.49-54
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• 1974

In the present studies, effect of progesterone on the germinal vesicle break-down of the mouse oocytes cultured in the micro tube was investigated. The results obtained are as follows: As dose of progesterone in the medium rose, accordingly the break-down of the germinal vesicle was suppressed. It was found that $ED_{50}$ was 15.7 ${\mu}g$/ml, and $ED_{90}$ 60.7 ${\mu}g$/ml of progesterone. The dose suppressing the oocyte maturation was apparently higher than that on the rabbit or on the mouse embryonal development. The inhibiting effect of progesterone on the GVBD was reversible. The germinal vesicle of the oocytes were broken down immediately in the medium upon removal of the hormone. Progesterone stops meiosis at any stage upon administration, while dbe AMP or theophylline supresses only the break-down of the nuclear membrane. Recovering of the meiotic division of the oocytes once exposed to progesterone was delayed a little. The inhibiting action of progesterone was not altered by adding more pyruvate or in the presence of higher concentration of the mineral ions in the culture medium.

• Clinical and Experimental Reproductive Medicine
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• 제45권4호
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• pp.154-162
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• 2018

Objective: The fallopian tubes play a critical role in the early events of fertilization. The rapid innate immune defense is an important part of the fallopian tubes. Toll-like receptor 3 (TLR3), as a part of the innate immune system, plays an important role in detecting viral infections. In this basic and experimental study, the effect of sex hormones on the function of TLR3 in the OE-E6/E7 cell line was investigated. Methods: The functionality of TLR3 in this cell line was evaluated by cytokine measurements (interleukin [IL]-6 and IL-1b) and the effects of sex hormones on TLR3 were tested by an enzyme-linked immunosorbent assay kit. Additionally, TLR3 small interfering RNA (siRNA) and a TLR3 function-blocking antibody were used to confirm our findings. Results: The production of IL-6 significantly increased in the presence of polyinosinic-polycytidylic acid (poly(I:C)) as the TLR3 ligand. Using a TLR3-siRNA-ransfected OE-E6/E7 cell line and function-blocking antibody confirmed that cytokine production was due to TLR3. In addition, 17-${\beta}$ estradiol and progesterone suppressed the production of IL-6 in the presence and absence of poly(I:C). Conclusion: These results imply that sex hormones exerted a suppressive effect on the function of TLR3 in the fallopian tube cell line when different concentrations of sex hormones were present. The current results also suggest that estrogen receptor beta and nuclear progesterone receptor B are likely to mediate the hormonal regulation of TLR3, as these two receptors are the main estrogen and progesterone receptors in OEE6/E7 cell line.

• 대한수의학회지
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• 제29권1호
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• pp.21-25
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• 1989

This experiment was carried out to determine the progesterone concentration of bovine plasma by liquid phase double antibody enzyme immunoassay. The optimum conditions of assay-system, enzyme conjugate and gelatin were invested. The sensitivity, recovery rate and reproducibility by this assay were also analyzed. The results obtained were as follows: 1. The suitable supplementation level of gelatin was 0.2%. As the gelatin level increased to 1%, coefficient variation of interassay was shown to be irregular. 2. The optimum dilution rate of enzyme conjugate was 30 times. Enzyme activity was greatly fluctuated depending on the minor condition of enzyme conjugate technique. Therefore, it was considered to be checked when determined. 3. The sensitivity of the assay was 12 pg/tube. 4. Recovery rate was decreased when the amount of sample was too little or too much, but the recovery rate was high as 97.8% when the amount of sample between 50 and $200{\mu}l$. 5. Mean intra-assay and inter-assay coefficient of variation was 4.5% and 5.9%, respectively. By using liquid phase double antibody enzyme immunoassay, progesterone in plasma can be detected, and also this assay system is applicable to study on physiological function of progesterone and to diagnosis of reproductive disorders.

• Asian-Australasian Journal of Animal Sciences
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• 제17권4호
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• pp.460-466
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• 2004

This study was undertaken with the aim to establish a reliable radioimmunoassay (RIA) system for urinary pregnanediol glucuronide (PdG) and to employ it for monitoring the reproductive status of dairy cows. Urine and blood samples were collected from the Holstein cows both pregnant and non-pregnant. The samples were then investigated for evaluating the relationship between progesterone ($P_{4}$) in blood and PdG in urine adjusted with or without urinary creatinine basis. Biweekly urine collection was employed for three cows in estrous and those artificially inseminated, while urine from pregnant cows was collected on a monthly basis. P_{4}$ and PdG levels were measured by enzymeimmunoassay (EIA) and RIA techniques, respectively. Our results indicated the sensitivity of PdG for RIA being 35 pg/tube and the recovery rate of 100%. Urinary creatinine concentrations also fluctuated within a day, but change at midday was not noteworthy. Regardless of the time of urination the change in concentrations of PdG was relatively smaller and did not vary significantly. The urinary PdG concentration showed periodic changes as that with serum P_{4}$ levels during the cow's estrus cycle. The correlation coefficient rose when creatinine level in urine was adjusted but the change was also not significant. The concentrations of PdG during the luteal phase were detected between 8.2 and 17.4 ng/ml, three to five times higher than that in the follicular phase. The concentration of PdG from pregnant cows (21 days after conception) was three to four times higher than in the nonpregnant cows. Our finding suggests that the determination of urinary PdG could be reliably employed for early pregnancy detection. The urinary PdG level continued to raise until 30 days pre-partum while the concentration reached its peak at 30 ng/ml, after which it started to fall 18 to 30 days before parturition and finally fell to its nadir value one week after parturition. As the correlation coefficient between the urinary PdG and serum P_{4}$ was higher than that corrected by urinary creatinine it can be suggested that the adjustment is not needed. The concentrations of urinary PdG could be maintained stably for 2 days in urine samples stored at room temperature and extended to 8 days when the samples were pretreated by boiling for 30 minutes. In conclusion urinary PdG concentration even without the need for creatinine basis adjustment can be used directly for monitoring the reproductive status of dairy cows.

• Asian-Australasian Journal of Animal Sciences
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• 제16권9호
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• pp.1254-1260
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• 2003

The purpose of this study firstly was conducted to establish a radioimmunoassay (RIA) of estrone sulfate ($E_1S$), secondly to monitor the reproductive status of dairy cows using their urine samples. Urine and blood samples were collected in series within a day from four pregnant Holstein-friesian cows to evaluate the relationship between $E_1S$ levels in blood and urine with or without urinary creatinine basis. The urine was then collected biweekly from three cows in estrous and those artificially inseminated; collection from pregnant cows was made on a monthly basis. Results indicated that sensitivity for the $E_1S$ RIA was 5 pg/tube and the recovery rate was 100%. The daily urinary creatinine concentrations fluctuated within a day, but changes were slighter in midday, whereas the changes of concentrations of $E_1S$ in urine were relatively smaller. The concentrations of serum $E_1S$ during the estrous cycle were undetectable due to the limitation of assay, but the urinary $E_1S$ level could be measured with no obvious changes during the cycle. The urinary $E_1S$ levels increased remarkably around 7.7 to 8.3 ng/ml, 80 to 100 days after pregnancy but the serum $E_1S$ levels did not elevate until 120 to 150 days. The level of $E_1S$ increased gradually during pregnancy and eventually reached its peak before parturition at around 40 ng/ml and finally decreased to its basal level 2 days postparturition. During pregnancy, $E_1S$ concentrations of urine increased earlier than those in blood. The correlation coefficients between urinary and serum $E_1S$ concentration during pregnancy and postparturm were higher than those adjusted with creatinine (creatinine ratio). The concentrations of $E_1S$ in urine could be maintained unchanged for 8 days storing the samples in room temperature, which was extended to 8 days when the samples were pretreated by boiling for 30 minutes or treated with autoclave. In conclusion urinary $E_1S$ concentrations can be used directly for monitoring the pregnant status and fetal viability of dairy cows and can assist accurate confirmation of pregnancy in cows at least 80 to 100 days after insemination much earlier than by serum $E_1S$.