The optimization of extraction conditions of phytoestrogen from pomegranate by hot water was conducted by analyzing the extraction characteristics. The purpose of this study was effective utilization of bioactive components of pomegranate, and the analyzing was performed with response surface methodology (RSM). This study established 10 sections based on the central composite design with the independent variables of extraction temperature (60, 70, 80, 90, $100^{\circ}C$) and extraction time (1, 2, 3, 4, 5 hr) to predict the optimal conditions for extraction of the effective components. The dependent variables were measured for extracted materials, those were, the major components such as chlorogenic acid, kaempferol, $17-{\alpha}-estradiol\;and\;17-{\beta}-estradiol$ content, and regression analysis was performed by SAS program, and optimal conditions for each characteristics were predicted, and the characteristics of extraction were analyzed by response surface methodology. It was found that chlorogenic acid, kaempferol, and $17-{\alpha}-estradiol$ content were greatly affected by extraction temperature. However, $17-{\beta}-estradiol$ content was affected significantly by extraction time. Regression formulas for each variable were elicited from this study, and the chlorogenic acid, kaempferol, $17-{\alpha}-estradiol\;and\;17-{\beta}-estradiol$ content depending on response surface methodology factor were superimposed. It was shown that optimal temperature and extraction time were $98{\sim}100^{\circ}C\;and\;3{\sim}5$ hrs, respectively.
The effects of exogenous estrogen on growth, mammary development, and the age at which the first estrous symptoms are detected were investigated for the first time in Korean native heifers at 6 months of age. At 90 days after treatment, weight gains of heifers implanted with 10 mg of estradiol were not different from those of the control heifers. However, weight gams of heifers implanted with 20 mg of estradiol were significantly increased at 90 days by 18%, relative to control values. At 180 days, cumulative weight gains of heifers implanted with either 10 mg or 20 mg of estradiol averaged 6.7% or 17.8%, respectively, greater than controls. But these values were not statistically significant. Regardless of dosage, teat diameter gains in the treatment groups were significantly increased at 90 days about 159%, relative to controls. Cumulative teat diameter gains in the heifers implanted with either 10 mg or 20 mg of estradiol were also significantly increased at 180 days by 100% or 128%, respectively, compared to controls. Teat length gains in the heifers implanted with either 10 mg or 20 mg of estradiol were significantly increased at 90 days by 200% and 295%, respectively, compared to controls. Cumulative teat length gains in the heifers implanted with either 10 mg or 20 mg of estradiol were significantly 265% and 325%, respectively, higher than controls. The heifers implanted with either 10 mg or 20 mg of estradiol showed a significant increase in teat volume gains at 90 days of 282% or 246%, respectively, over those of the control heifers. Cumulative teat volume gains in the heifers implanted with either 10 mg or 20 mg of estradiol were significantly increased at 180 days by 251% and 244%, respectively, compared to control values. However, there were no significant differences m gams of teat diameter, length and volume between heifers implanted with 10 mg and 20 mg of estradiol regardless of 90 days or 180 days. Heifers implanted with 10 mg or 20 mg of estradiol showed the first estrus 20 or 124 days, respectively, faster than controls. Overall, These data strongly suggest that estradiol implantation into heifers stimulates growth and mammary development and hasten the age at which the first estrus in Korean native heifers is observed.
It has been well known that ovarian steroids, estradiol and progesterone play an important role in the endometrial preparation for the implantation process. The present investigation was undertaken to correlate function of ovarian steroids with cAMP concentrations in uterine tissues of rat during the various stages of the preimplantation period. Rats ovariectomized on day 2 were treated with estradiol or/and progesterone on day 3 or on day 4 and cAMP concentrations in uterine tissues were determined by competitive protein - binding assay in control - and steroid treated - ovariectomized rats. The results obtained are as follows; 1. In control rats, cAMP concentrations in uterine tissues were decreased with preimplantation period proceeded whereas cAMP concentrations were increased and showed the highest levels on day 6 in ovariectomized rats. 2. In rats treated with progesterone only or progesterone with estradiol after ovariectomy, cAMP concentrations on day 6 were lower than those of ovariectomized control but higher than those of intact control rats while estradiol only treatment decreased cAMP concentrations on day 3 and day 6, compared with those from intact- and ovariectomized-controls. It is, therefore, concluded that the concentrations of cAMP in uterine tissues were lower in estrogen-treated rats than in ovariectomized rats, suggesting the involvement of cAMP in the regulation of uterine tissue differentiation.
Neuronal apoptotic events, which result in cell death, are occurred in hypoxic/ischemic conditions. Estradiol is a female sex hormone with steroid structure known to provide neuroprotection through multiple mechanisms in the central nervous system. This study was aimed to investigate the signal transduction pathway of $CoCl_2$-induced neuronal cell death and the inhibitory effects of estradiol. Administration of $CoCl_2$ decreased cell viability in both a dose- and time-dependent manner in PC12 cells. $CoCl_2$-induced cell death produced genomic DNA fragmentation and morphologic changes such as cell shrinkage and condensed nuclei. It was found that $CoCl_2$-treated cells increased the reactive oxygen species (ROS) as well as caspase-8, -9 and -3 activities. However, pretreatment with estradiol before exposure to $CoCl_2$ prevented the reduction in cell viability reduction and attenuated DNA fragmentation and morphologic changes caused by $CoCl_2$. Furthermore, the $CoCl_2$-induced increases of ROS levels and caspases activities were attenuated by estradiol. Gene expression analysis revealed that estradiol blocked the underexpression of the Bcl-2 and ameliorated the increase in the release of cytochrome c from mitochondria into cytoplasm and Fas-ligand (Fas-L) upregulated by $CoCl_2$. These results suggest that $CoCl_2$ induce apoptosis in PC12 cells through both mitochondria- and death receptor-mediated cell death pathway. Estradiol was found to have a neuroprotective effect against $CoCl_2$-induced apoptosis through the inhibition of ROS production and by modulating apoptotic effectors associated with the mitochondria- and death-dependent pathway in PC12 cells.
Kim, Young-Seob;Kim, Ji-Yong;Jung, So-Young;Lee, Bong-Joo;Shin, Nam-Shik
Journal of Veterinary Clinics
/
v.27
no.6
/
pp.693-697
/
2010
This study, conducted with four leopard cats (Prionailurus bengalensis), used time-resolved fluoroimmunoassay (TR-FIA) to analyze estradiol and progesterone concentrations in fecal samples. We measured fecal samples taken during estrus period, diestrus period, pregnancy and non-pregnancy period. During estrus (February), the mean minimum estradiol concentration was $4.02{\pm}1.9$ng/g, and the mean maximum was $86.01{\pm}35.2$ng/g (dry fecal weight). During diestrus (November), the mean minimum estradiol concentration was $4.42{\pm}1.32$ng/g and mean maximum was $15.62{\pm}6.48$ng/g (dry fecal weight). Midgestation (April), the mean minimum progesterone concentration was $427{\pm}24.49$ng/g and the mean maximum was $1490{\pm}265.27$ng/g. During non-pregnancy (November), the mean minimum progesterone concentration was $71.25{\pm}29.61$ng/g and the mean maximum was $291.75{\pm}90.30$ng/g. These results suggest that steroid hormone analysis of feces using TR-FIA is a valid method for noninvasively determining ovarian activity associated with estrus and pregnancy in leopard cats. This study will contribute to building breeding management and reproductive plans for endangered species.
The study was carried out to find out the changes of the sex hormone concentrations in the blood plasma and antrum of follicular and lutein cystic ovaries of Holstein cows. The progesterone, estradiol-17$\beta$, testosterone, FSH and LH from samples of the blood plasma and antrum of cystic ovaries of cows assayed by radioimmunoassay method. The results of this study were summarized as follows : 1. The concentrations fo progesterone and estradiol-17$\beta$ in the blood plasmaat estrous and luteal phase were 0.95$\pm$0.18ng/ml, 11.45$\pm$3.12pg/ml and 4.25$\pm$0.27ng/ml, 6.27$\pm$0.82pg/ml respectively. The concentrations of progesterone and estradiol-17$\beta$ in the antrum fluid of follicles at estrous and luteal phase were 24.8$\pm$4.12ng/ml, 54.3$\pm$7.25pg/ml and 21.7$\pm$3.79ng/ml, 14.3$\pm$2.72pg/ml respectively, and showed significant changes among the estrous and luteal phase and blood plasma and antrum fluid of follicles. 2. The concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the blood plasma of follicular cystic ovareis of cows were 0.85$\pm$0.25ng/ml, 9.23$\pm$2.72pg/ml, 17.12$\pm$3.26pg/ml and 3.78$\pm$1.02mIU/ml respectively. And the concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the antrum fluid of follicular cystic ovareis of cows were 284$\pm$48.21ng/ml, 389$\pm$67.23ng/ml, 12.84$\pm$0.29ng/ml and 1.84$\pm$0.17mIU/ml respectively, and showed significant changes between in the blood plasma and antrum fluid of cystic ovaries. 3. The concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the blood plasma of lutein cystic ovaries of cows were 3.40$\pm$0.78ng/ml, 4.02$\pm$0.42pg/ml, 10.72$\pm$2.74pg/ml and 0.76$\pm$0.12mIU/ml respectively. And the concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the antrum fluid of lutein cystic ovareis of cows were 427$\pm$35.79ng/ml, 0.76$\pm$0.07ng/ml, 3.45$\pm$0.57ng/ml and 0.29$\pm$0.07mIU/ml respectively, and showed significant changes between the blood plasma and antrum fluid of cystic ovaries. 4. Accordingly, the diagnosis of follicular and lutein cystic ovareis of cows from progesterone, estradiol-17$\beta$ and LH levels in the blood plasma and antrum of cystic ovaries of cows is thought to be possible a diagnostic means.
Kim, Jeong-Bae;Kim, Bang-Sil;Mun, Byeong-Gwon;Yun, Chang-Jin;Park, Chul-Ho;Moon, Jin-San;Suh, Guk-Hyun;Oh, Ki-Seok;Son, Chang-Ho
Journal of Veterinary Clinics
/
v.25
no.2
/
pp.79-84
/
2008
For estimating the ovulation time in Miniature Schnauzer dogs during the estrous cycle, radioimmunoassay of plasma estradiol-$17{\beta}$ and progesterone concentrations was conducted on blood samples in 21 pregnant and 13 non pregnant dogs. When Day 0 was that plasma progesterone concentrations exceeded 4.0 ng/ml, on Day 64, parturition day, progesterone declined below 1.0 ng/ml with $0.92\;{\pm}\;0.29\;ng/ml$ and when Day 0 was that plasma progesterone concentrations declined below 1.0 ng/ml, on Day -64, progesterone increased above 4.0 ng/ml with $4.56\;{\pm}\;0.87\;ng/ml$. Gestational length was $63.71\;{\pm}\;1.35$ (Mean${\pm}$S.D.) days from plasma progesterone concentrations exceeded 4.0 ng/ml and was $66.29\;{\pm}\;1.98$ days from first male acceptance. The plasma estradiol-$17{\beta}$ concentrations reached maximum value with $28.20\;{\pm}\;2.86\;pg/ml$ on Day .2, and plasma progesterone concentrations reached $5.90\;{\pm}\;0.36 ng/ml, 5.18\;{\pm}\;0.32 ng/ml on Day 0, and the maximum of 61.58\;{\pm}\;10.47 ng/ml on Day 19 and 56.05\;{\pm}\;8.86\;ng/ml$ on Day 16 in pregnant and non pregnant dogs, respectively. Afterward, plasma progesterone concentrations declined below 1.0 ng/ml on Day 64 with $0.92\;{\pm}\;0.29\;ng/ml$ in pregnant cycles and on Day 58 with $0.95\;{\pm}\;0.63\;ng/ml$ in non pregnant dogs. No difference were found pregnant and non pregnant dogs in plasma estradiol-$17{\beta}$ and progesterone concentrations (p<0.01). Based on first male acceptance (Day 0), the maximum of plasma estradiol-$17{\beta}$ concentrations ($29.31\;{\pm}\;3.61\;pg/ml$) occurred on Day -1 and plasma progesterone concentrations exceeded 4.0 ng/ml on Day 2 in pregnant ($5.37\;{\pm}\;0.76\;ng/ml$) and non pregnant ($4.25\;{\pm}\;0.80\;ng/ml$) dogs. These results suggest that in Miniature Schnauzers, the ovulation occurred when plasma progesterone concentrations exceeded 4.0 ng/ml, 3 days after plasma estradiol-$17{\beta}$ peak and 2 days after first male acceptance.
Pang, X.S.;Wang, Z.Y.;Zhu, T.G.;Yin, D.Z.;Zhang, Y.L.;Meng, L.;Wang, F.
Asian-Australasian Journal of Animal Sciences
/
v.23
no.2
/
pp.188-196
/
2010
The objective of this study was to characterize the litter sizes in Huanghuai goats with high prolificacy (HP, five or more kids born per litter on at least one occasion), and to compare their peripheral blood concentrations of progesterone and estradiol with those of goats with poor prolificacy (PP, up to three kids born per litter on any occasion). The circulating concentrations of progesterone and estradiol were measured by radioimmunoassay from daily blood samples taken during natural estrus cycles and at 1-5 days after ovariectomy. Estrus was synchronized using two doses of a prostaglandin analog. Litter size for the HP goats increased up to a parity of five and decreased thereafter. The percentage of goats with litter sizes of $\geq$4 from parities 3 to 6 ranged from 44.5% to 58.3%. Although small differences in litter size were obtained for goats of parities three, four and six relative to five, parity five does had the highest mean litter size. Progesterone concentrations began to rise earlier and were higher in the HP than in the PP goats on most days of the luteal phase, but not during the follicular phase of the cycle or after ovariectomy. There was a significant difference between the two groups (p<0.05) in the magnitude of the progesterone plateau. Mean estradiol concentrations in the HP group remained significantly higher than in the PP group (p<0.05) during the estrus cycle. There were two estradiol peaks in the HP goats during the early luteal phase, but only one in the PP group. Measurements of individual corpora lutea (CL) in vitro showed that there was a greater prevalence of small CL (<6 mm in diameter) in the HP group than in the PP group (p<0.05). After ovariectomy, the estradiol level on day 1 was significantly higher than at the nadir during the estrus cycle in both the HP (p<0.01) and PP (p<0.05) goats, while levels decreased by 12.3% and 26.2% respectively compared with the mid-luteal period in HP and PP goats (p<0.05). The overall mean estradiol concentrations in HP goats were lower than in the PP group, but no significant differences were found between groups at 1-5 days after ovariectomy.
The present study was carried out to establish a practical regimen for artificial parturition induction using prostaglandin F2$\alpha$(PGF2$\alpha$) and estradiol-benezoate in Korean native goats. The effect of parturition induction and the time intervals to induced parturition after injection were investigated. The birth weight and body weight of kids at 15 days of age were measured. A total of 24 pluriparous goats were offered for this experiment. The animals were divided into 4 goats per treatment by the injection time(142, 145 or 148 day of pregnancy) and dosage(5.0$\times$10 or 7.5$\times$7.5mg). The results obtained were summarized as follows : A total of 24 pregnant goats were intramusculary treated with 5.0$\times$10 or 7.5$\times$7.5mg of PGF2$\alpha$ and estradiol-benzoate for parturition induction of Day 142, 145 or 148 of gestation. Parturition was induced in all of the goats(100%) treated. The kids produced from induced parturition were all healthy. The time intervals to induced parturition after PGF2$\alpha$ and estradiol-benezoate injection of 5.0$\times$10 or 7.5$\times$7.5mg to pregnant goats on Day 148(23.22$\pm$0.51~23.40$\pm$1.26hrs) were significantly(P<.01) shorter than those of the 142 days of the gestation(26.34$\pm$2.22~28.39$\pm$3.02hrs). No significant difference was found in the time intervals between the doses(5.0$\times$10 or 7.5$\times$7.5mg) treated for parturition induction. The birth weight of kids from induced parturition was no significant difference between on Day 148 and on Day 142 of gestation. However, the birth weight of kids from parturition induced on Day 148 was found significantly(P<.01) heavier than that of the 142 days of gestation. The body weight of kids at 15 days old was also significantly(P<.01) lighter in the parturition induced on day 142 than those on Day 142. The birth weight and body weight of kids at 15 days old were not affected by 5.0$\times$10 or 7.5$\times$7.5mg injection of PGF2$\alpha$ and estradiol-benzoate for inducing parturition. From the above results, it was concluded that the parturition induction by PGF2$\alpha$ and estradiol-benezoate injection of 5.0$\times$10 or 7.5$\times$7.5mg on Day 142 of gestation, which was correspondent to 8 days before expected spontaneous parturition, was available without any significant troubles.
Seo, Kil Woong;Kim, Chong Sup;Park, Chang Sik;Lee, Kyu Seung
Korean Journal of Agricultural Science
/
v.16
no.2
/
pp.169-178
/
1989
The study was carried out to elucidate the feedback mechanism on the hypothalamo-hypophyseal system from the functional changes of ovary in female rats. One hundred and forty-four mature female rats were lloted into the three groups; ovariectoimzed group, estradiol treated group and intact control group. The varies of 48 heads of rat were completely removed. Forty eight heads of rat were administered with $200{\mu}g$ of estradiol benzoate every 48 hours. Serum FSH, LH and prolactin levels were determined with radioimmunoassay method at 3,6,12,24 ours, and 5,10, and 15days after treatment. The rats were necropsied to measure the weights of hypophysis and to examin the histological changes in the organs. The results obtained were as follows: The weights of hypophysis were increased after ovariectomy and decreased after estradiol injection. The differences in hypophysis weights were significant between the group from 5 days after treatment. The histological changes in hypophysis were appeared from 5th day after ovariectomy. Proliferation and hypertrophy began to occur in basophilic from 10th day after ovariectomy, chromophobes were slightly hypertrophied and acidophilic cells were atrophied. In estradiol injected rats the histological findings were appeard to be contrary to those of ovariectomized rats. Serum FSH levels significantly changed after ovariectomy and estradiol injection and were higher in both the treated groups than in the intact control group. Within 18 hours after treatment the level was the highest in ovariectomized group, and thereafter the highest level was found in estradiol treated gorup. In ovariectomized rats the levels were rapidly increased 3 hours after treatment and maximum levels were found 18 hours after treatment. In estradiol treated rats the levels started to increase 18 hours after treatment and reached maximum levels 24 hours treatment. 4. Serum LH levels started to increase 3 hours after ovariectomy and estradiol injection and reached maximum levels 12 hours after ovariectomy and 24 hours after estradiol injection. There were significant differences in LH levels between the groups in each observation time. Up to 18 hours after treatment levels were higher in ovariectomized rats than in estradiol treated rats. but thereafter the levels were higher in estradiol treated rats than in ovariectomized rats. The multiple range test showed that a significant difference in LH levels was not found between ovariectomized group and estradiol treated group 18 hours and 5 days after treatment. 5. Serum prolactin levels were significantly changed after ovariectomy and estradiol injection. The levels were lower in ovariectomized rats than in intact control rats.
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