• Journal of Veterinary Science
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• v.24 no.1
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• pp.4.1-4.16
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• 2023

Background: In vitro culture of preantral follicles is a promising technology for fertility preservation. Objectives: This study aims to investigate an optimized three-dimensional (3D) fetal bovine serum (FBS)-free preantral follicle culture system having a simple and easy operation. Methods: The isolated follicles from mouse ovaries were randomly divided in an ultra-low attachment 96-well plates supplement with FBS or bovine serum albumin (BSA) culture or encapsulated with an alginate supplement with FBS or BSA culture. Meanwhile, estradiol (E₂) concentration was assessed through enzyme-linked immunosorbent assay of culture supernatants. The diameter of follicular growth was measured, and the lumen of the follicle was photographed. Spindle microtubules of oocytes were detected via immunofluorescence. The ability of oocytes to fertilize was assessed using in vitro fertilization. Results: The diameters were larger for the growing secondary follicles cultured in ultra-low attachment 96-well plates than in the alginate gel on days 6, 8, and 10 (p < 0.05). Meanwhile, the E2 concentration in the BSA-supplemented medium was significantly higher in the alginate gel than in the other three groups on days 6 and 8 (p < 0.05), and the oocytes in the FBS-free system could complete meiosis and fertilization in vitro. Conclusions: The present study furnishes insights into the mature oocytes obtained from the 3D culture of the preantral follicle by using ultra-low attachment 96-well plate with an FBS-free system in vitro and supports the clinical practices to achieve competent, mature oocytes for in vitro fertilization.

• Proceedings of the KSAR Conference
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• 2005.06a
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• pp.47-54
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• 2005

In boars, unlike the cases in males of other species, gonadal hormones suppress voluntary feed intake. for this reason, barrows, compared with gilts or boars, eat too much feed resulting in excessive fat deposition. Two experiments were performed in the present study to investigate the effects of implantation of Revalor H[Experiment(Exp.) I: 140mg trenbolone acetate(a synthetic androgen) + 14mg estradiol-$17\beta(E_{2}\beta)$] and Compudose(Exp. II; 24mg $E_{2}\beta$) on growth efficiency, carcass characteristics and circulating concentrations of IGF-I and IGF-binding protein-3(IGFBP-3). In Exp. I, sixty-four cross-bred finishing barrows weighing approximately 60kg were randomly divided into eight pens under a 2[control vs Revalor implant] $\times$ 2(ad libitum vs $80\%$ ad libitum feeding) $\times$2[control($103\%$ NRC-recommended level) vs low-energy($87\%$ NRC recommendation) diet] arrangement of treatments. In Exp. II, effects of Compudose were studied using 80 finishing barrows(10 animals/pen). In both Exps., all the animals were slaughtered at 100- to 110-kg body weight. Both Revalor and Compudose implants caused a decrease in feed intake and backfat thickness without affecting major physicochemical characteristics of the carcass and an increase in circulating IGF-I concentration. Moreover, Revalor implant exhibited greater effects than restricted feeding, low-energy diet, or Compudose in these variables. In addition, Revalor implantation suppressed weight gain, but enhanced the feed efficiency without exhibiting any interaction with the diet or feeding. In summary, results suggest that 1) both androgen and estrogen suppress voluntary feed intake and backfat deposition and enhance IGF-I secretion and 2) these effects of the gonadal steroid hormones in growth are likely to be mediated, in part, by IGF-I in finishing barrows.

• Journal of Applied Biological Chemistry
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• v.59 no.4
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• pp.305-311
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• 2016

This study investigated the effects of a mixture of fenugreek seeds and Lespedeza cuneata extracts on testosterone synthesis in TM3 cells that were oxidatively stressed with $H_2O_2$. In order to oxidatively stress TM3 cells, the cells were treated with $50{\mu}M$ hydrogen peroxide for 4 hr in serum-free media. Yagwanmun-horopa mixture (YHM) showed neither cytotoxicity nor increment of cell proliferation in the oxidatively stressed TM3 cells in any concentration. When the cells were treated with hydrogen peroxide, testosterone levels decreased, but the testosterone level was returned to that of the control level in the presence of YHM. In order to find out the reasons for the increase of testosterone, the expression of the genes involved in the synthesis or disintegration of testosterone. On the other hand, the levels of $3{\beta}$-HSD4 and 17, 20-desmorase, which are involved in testosterone synthesis, were decreased through the use of hydrogen peroxide and were recovered through YHM treatment. Aromatase and $5{\alpha}$-reductase2, which convert testosterone to estradiol and dihydrotestosterone, respectively, were increased through the use of hydrogen peroxide, and were returned to control level through YHM treatment. These results suggest that YHM does not affect TM3 cell proliferation. However, YHM increases the expression of testosterone-synthesizing enzyme, which was decreased through oxidative stress, and decreases the expression of testosterone- converting enzyme, which was increased through oxidative stress. Therefore, it is reasonable that YHM has strong recovery activity on testosterone to normal level, even in the oxidatively stressed TM3 cells which mimics the andropause state.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.18 no.3
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• pp.224-238
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• 2008

The aim of this study is to confirm the physicochemical property and hazard of thinner (012), which is a diluent of enamel paint used for floor coating for waterproofing and oil painting for the outer wall. The literatures of physicochemical property and hazard of thinner were surveyed and its physicochemical property were evaluated. And then, the inhalation toxicity of thinner affecting the central nervous system and reproductive organs in rats were examined by subchronic (6 h./day. 5 days/ week for 13 weeks) inhalation test. 1) According to the 13-week subchronic inhalation test, there were no significant changes in clinical test and body weight. However, a significant evidence of toxicity was observed in the hematological test and organ weight such as heart, kidney, liver and brain (p<0.01) in the 200 ppm and 1,000 ppm exposure groups in a dose response manner. In the histopathology analysis, there were no significant evidence of toxicity. Therefore, thinner was not classified as an organ targeted toxic agent. In case of Harmfulness, it could be classified as a chronic toxic agent 3($500 ppm/4hr, rat). 2) The reproductive toxicity such as extension of the period of estrous cycle, reduction of serum estradiol concentration and increase of frequency of the abnormal sperm was observed in the 1,000 ppm exposed animals. 3) The result of the physicochemical property of the test material showed that the specific gravity was 0.793, boiling point $155.8^{\circ}C$, steam pressure 2.1 kPa, ignition point $34.5^{\circ}C$, and spontaneous ignition point $280^{\circ}C$. The endothermic and exothermic values were 371.4 J/g and 159.1 J/g. respectively. The explosion limit was 214 mg/l. These data showed that thinner could be classified as an explosion agent level 1.2 and ignitive liquid agent 3 ($23-60^{\circ}C$) according to the notification No. 2008-1 of the Labor Ministry, "Classifying Standard of Chemical Materials."

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.8
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• pp.1080-1088
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• 2010

This experiment was conducted to investigate the effects of dietary energy levels of gestating gilts on physiological parameters and reproductive performance for primiparous sows. A total of 40 F1 gilts (Large White${\times}$Landrace) were allocated to 4 treatments using a completely randomized design (CRD). Four different experimental diets contained 3,165, 3,265 3,365 and 3,465 kcal of ME/kg and each diet was provided to gilts at 2.0 kg/d during gestation. Consequently, energy intake of each treatment of gestating gilts was 6,330, 6,530, 6,730 and 6,930 kcal ME/kg, respectively. During the whole gestation period, body weight, fat mass gain and backfat thickness of gilts were increased in proportion to dietary energy levels (p<0.01). However, estimated protein mass gain of gilts was not affected by dietary energy level (p>0.10). At farrowing, the total number of pigs born per litter did not show any significant difference among treatments. However, the number of pigs born alive per litter in treatment 6,730 kcal ME/d was significantly higher than that of other treatments (p<0.05). Moreover, litter weight at birth was improved as dietary energy level was increased (p<0.05). Feed intake of sows during lactation tended to decrease as dietary energy level of gestation was increased, but litter weight gain was not affected by dietary treatment during the gestation period. Fat content in colostrum was higher as dietary energy level was increased during gestation. The concentration of blood estradiol-$17{\beta}$ was increased and was higher at the first trimester of gestation in 6,730 kcal ME/d treatment compared to other treatments. These results suggested that increased dietary energy level during gestation resulted in higher body weight and backfat thickness of sows. In addition, reproductive performance of the sow, such as litter weight at farrowing and the number of pigs born alive, was improved when 6,730 kcal of ME/d treatment diet was provided. Consequently, the NRC (1998) recommendation of energy for gestating gilts (6,015 to 6,150 kcal of ME/d) should be reevaluated to maximize reproductive performance because recent high-producing sows require much more energy to produce a large litter size and heavier piglets from the first parity.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.117-121
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• 2008

Our previous study showed that transgenic (TG) pigs harboring human EPO (hEPO) gene have been shown to have reproductive disorders, including low pregnancy rates, irregular estrus cycle and low little size. To investigate these reasons, we assessed estrus behavior (standing response) and plasma $17{\beta}$-estradiol ($E_2$) level, which partly reflect reproductive function, during the estrus cycles after synchronization and superovulation by hormone treatments. Then, we analysed blood composition and expression of hEPO gene in TG pigs. Pigs were injected with PG600. After 10 days, pigs were fed with Regumate porcine for 6 days. Blood samples were collected from jugular vein. Analysis of blood composition and $E_2$ level were measured by Hemavet 950 and $E_2$ ELISA kit, respectively. And, the expression of hEPO gene in reproductive organs was quantitated by real-time RT-PCR. The percentage of estrus behavior in TG was significantly decreased. Hematocrit (HCT), hemoglobin (Hb) concentration and red blood cell (RBC) number were significantly higher in TG than wild type (WT). On the other hand, high expression of hEPO gene in TG was observed in the mammary gland as well as in the uterus. Moreover, plasma $E_2$ level was significantly higher in TG than WT. These results suggest that nonspecific expression of hEPO gene in the other organs of TG may affect blood composition and plasma $E_2$ level, thereby causing reproductive disorders.

• Fisheries and Aquatic Sciences
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• v.12 no.4
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• pp.293-298
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• 2009

Several studies have reported that nonylphenol (NP) and 2,2', 4,6,6'-pentachlorobiphenyl (PCB104) exhibit estrogenic activity. To investigate the estrogenic potency of NP and PCB104 during oocyte maturation, fully vitellogenic oocytes (0.76 mm diameter in average) of yellow fin goby, Acanthogobius flavimanus, were exposed in vitro to these chemicals at different concentrations (0.1, 1, 10, 100, and 1,000 ng/mL) with the exogenous precursor $17\alpha$-hydroxyprogesterone ($17{\alpha}OHP$) 50 ng/mL in the presence or absence of human chorionic gonadotropin (HCG). The production of testosterone (T), estradiol-$17\beta$ (E2), and $17\alpha,20\beta$-dihydroxy-4-pregnen-3-one ($17\alpha20{\beta}OHP$) in response to NP or PCB104 were measured by radioimmunoassay. Steroid levels were also expressed as E2/T and E2/$17\alpha20{\beta}OHP$ ratios. In the absence of HCG, no significant differences in either NP or PCB104 treatment groups were observed. In the presence of HCG, NP treatment did not show significant differences in the production of T, E2, and $17\alpha20{\beta}OHP$ at any concentrations tested, but E2/T ratios were decreased at concentrations of 0.1, 1, 10, and 1,000 ng/mL compared with the control group. PCB104 decreased E2 production at concentrations of 0.1, 10, and 1000 ng/mL, but did not show significant differences in the production of T and $17\alpha20{\beta}OHP$ at any concentration tested. While E2/T ratios were decreased at PCB104 concentrations of 0.1, 1, 10, and 1,000 ng/mL, E2/$17\alpha20{\beta}OHP$ ratios were also decreased at 0.1, 10, and 1,000 ng/mL compared with the control. Results indicate that both NP and PCB104 appeared to have antiestrogenic effects during this phase.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2003.10a
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• pp.57-69
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• 2003

It is well known that many pesticides possess hormonal activity, and affect the developments of wildlife and mammals including human. Currently, pyrethroid insecticides are in worldwide use to control in and outdoor pests, providing potential far environmental exposure. Hormonal activities of these pyrethroid insecticides, however, have been little studied, and the developmental effects of them were no reported. Therefore, we firstly examined the potential estrogenic activities of some pyrethroid insecticides (permethrin, cypermethrin, tetramethrin, deltamethrin, sumithrin, fenvalerate and bioallethrin) by immature rat uterotrophic assay, luciferase reporter gene assay and Calbindin-D$\sub$9k/ (CaBP-9k) gene expression assay. Uterine wet weights were increased by permethrin and the permethrin-induced weights were inhibited by ICI 182780 in the uterolrophic assay. On the other hand tetramethrin significantly reduced uterine and vaginal wet weights, and also inhibited the E2-induced weight increases at all doses tested. Cypermethrin and sumithrin had a tendency to increase uterine weights, although not statistically significant. Permethrin and cypermethrin dose-dependently increased the luciferase activity in reporter gene assay. Northern blot analysis showed that permethrin induced CaBP-9k mRNA expression whereas tetramethrin inhibted. Subsequent studies were conducted to investigate the possible developmental effects of four pyrethroid insecricides (permethrin, cypermethrin, sumithrin and teramethrin). Either diethlbestrol (DES) or 17${\beta}$ -estradiol (E2) was used as a reference control in this study. Pyrethroid insecticides were administered to Sprague Dawley rats via subcutaneous injection at 6 to 18 days of gestation or 1 to 5 days after birth. In utero treatment of permethrin (10mg/kg/day) in female rat resulted in significant increases in uterine and ovarian weights while significant decreases in serum E2 concentration, uterine and ovarian ER${\alpha}$ mRNA levels. Sumithrin and permethrin led to acceleration in vaginal opening of female rat, while delay in preputial separation of male after neonatal treatment. Anogenital distances of PND 18 were significantly reduced in sumthrin-treated, and permerhrin-treated male rats after neonatal treatment. All the pyrethroid insecticides tested caused significant increases in uterine weights on PND 18, while significant reductions in the first diestrus phase when neonataly treated. In addition, exposure to pyrethroids in neonatal period led to significant reduction in relative brain weight in female rat on PND 18, but its weight was recovered in diestrus phase. In summary, Our experimental data demonstrate the possibilities of developmental effects of pyrethroid insecticides via estrogenic or antiestrogenic activity.

• Journal of Fisheries and Marine Sciences Education
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• v.24 no.6
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• pp.845-853
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• 2012

To investigate the role of temperatures and photoperiods as environmental cues regulating reproductive rhythm in Pseudobagrus fulvidraco, rearing experiments were conducted using sveral rearing regimes conbined with photoperiods and water temperatures during gonadal degeneration periods. Gonadosomatic index (GSI) in control was $8.16{\pm}1.50%$, while in other experiment GSI levels in female were lower than that in the control. In case of experimental precinct of 9 light (L) and 15 L, GSI levels were decreased. But GSI level with $20^{\circ}C$ was no diffrence after 40 and 60 days. GSI level in male of control was $0.35{\pm}0.05%$. GSI under 9 L and $25^{\circ}C$ was similar to that in control, whereas its level in other experiments was lower than that in control. Testosterone (T) of female was $3.68{\pm}0.22$ ng/mL at experimental precinct. In case of 9 L and 15 L, concentration of T were lower than experimental precinct in all of water temperature. Estradiol-$17{\beta}$ ($E_2$) and $7{\alpha}$, $20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}OHP$) levels of female were $0.42{\pm}0.02$ and $0.83{\pm}0.01$ ng/mL at experimental precinct. $E_2$ levels of 9 and 15 L were higher than experimental precinct and $17{\alpha}20{\beta}OHP$ levels of 9 and 15 L were higher than experimental precinct. In case of T and 11-ketotestosterone levels were $0.69{\pm}0.11$ and $.62{\pm}0.03$ ng/mL in male. During the period of gonadal degeneration, gonadal maturation did not occur in any of the experimental regimes. However, comparatively high levels of $E_2$ observed at low temperature regimes ($20^{\circ}C$) regardless of photoperiods.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.2
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• pp.167-177
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• 1997

Oocyte donation program developed to reach the pregnancy in those patients suffering from premature ovarian failure or surgery induced menopause, particularly in their reproductive age. With technical advances and popularity of ART (assisted reproductive technology), the indication of oocyte donation program extended to low responders, and even to naturally menopaused patients that has led them quite successfully to getting in pregnancy. The purpose of this study was to evaluate which one is involved in the decline of fertility between the oocyte and uterine factor. One hundred five cycles of oocyte donation program were performed in 84 patients from Jan., 1993 to Dec., 1996. Oocytes were donated from healthy, young, fertile anonymous donors or relatives or infertile patients with supernumerary oocytes. The study population was divided into 3 groups according to the age of recipients. Group 1 was less than 35 years old, Group 2 was between 35 to 39 years old, and Group 3 was more than 39 years old. The results were as follows: The mean age of oocyte donor was $31.5{\pm}3.3$ (range; 25-36). The mean concentration of basal serum FSH and peak serum estradiol were not different among groups. The mean number of oocytes retrieved from donors, embryos transferred to recipients, and fertilization rate were not different among groups. The clinical pregnancy rate was 37.3% in Group 1, 31.6% in Group 2, and 31.6% in Group 3, respectively. The spontaneous abortion rate was 16.0% in Group 1, 16.7% in Group 2, and 16.7 in Group 3, respectively. The multiple pregnancy rate was 20.0% in Group 1, 16.7% in Group 2, 16,7% in Group 3, respectively, The implantation rate was 11.3% in Group 1, 10.3% in Group 2 and 10.0% in Group 3, respectively. All of the pregnancy outcomes were not different statistically among groups. In conclusion, endometrial receptivity does not seem to be impaired as age increases with transfer of good quality embryos and adequate endometrial preparation.