• Journal of The Korean Society of Inherited Metabolic disease
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• v.14 no.2
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• pp.150-155
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• 2014

The measurement of $17{\alpha}$-hydroxyprogesterone ($17{\alpha}$-OHP) in a dried blood spot on filter paper is an important for screening of congenital adrenal hyperplasia (CAH). Since high levels of $17{\alpha}$-OHP are frequently observed in premature infants without congenital adrenal hyperplasia, we evaluated cuts-off based on birth weight and performed validation. Birth weight and $17{\alpha}$-OHP concentration data of 292,204 newborn screening subjects in Greencross labopratories were analyzed. The cut-off values based on birth weight were newly evaluated and validated with the original data. The mean $17{\alpha}$-OHP concentration were 7.25 ng/mL in very low birth weight (VLBW) group, 4.02 ng/mL in low birth weight (LBW) group, 2.53 g/mL in normal birth weight (NBW) group, and 2.24 ng/mL in heavy birth weight (HBW) group. The cut-offs for CAH were decided as follows: 21.12 ng/mL for VLBW and LBW groups and 11.14 ng/mL for NBW and HBW groups. When applied new cut-offs for original data, positive rates in VLBW and LBW groups were decreased and positive rates in NBW and HBW groups were increased. The cut-offs based on birth weight should be used in the screening for CAH. We believe that our new cut-off reduce the false positive rate and false negative rate and our experience for cut-off set up and validation will be helpful for other laboratories doing newborn screening test.

• Journal of Aquaculture
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• v.9 no.1
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• pp.57-63
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• 1996

The relative effectiveness of steroids and human chorionic gonadotropin (HCG) on germinal vesicle breakdown (GVBD) was investigated in vitro using the isolated oocytes (folliculated oocytes) from the spotted halibut, Verasper variegatus. Among the steroids tested, $17\alpha-hydroxy,\;20\beta-dihydroprogesterone\;(17\alpha20{\beta}OHP)$ was more effective than progesterone (P4) and $17\alpha-hydroxyprogesterone\;(17{\alpha}OHP$). In comparing $17\alpha,\;20{\beta}OHP$ with HCG, both $17\alpha20{\beta}OHP$ and HCG were effective in inducing GVBD at all concentrations : $17\alpha20{\beta}OHP$ was effective even at lower concentrations ($10\~50$ ng/ml). HCG at three concentrations used (50, 100, 500 IU/ml) showed no significant differences in inducing GVBD.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.2
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• pp.203-210
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• 1997

The aim of this study was to determine the effect of steroids and human chorionic gonadotropin (HCG) on in vitro maturation and ovulation of oocyte in Pseudobagrus fulvidraco. Oocytes were incubated in the media Leibovitz L15 supplemented with the various concentration of $17\alpha,\;20\beta-dihydroxy-4-pregnen-3-one(17\alpha20{\beta}OHP),\;17\alpha-hydroxyprogesterone(17{\alpha}OHP),\;progesterone(P_4),\;estradiol-17\beta(E_2)and\;HCG$. After 60 hours incubation, the maturation ability of oocyte was assessed by the appearance of germinal vesicle breakdown (GVBD). GVBD was significantly enhanced by the addition of $17\alpha20{\beta}OHP,\;17{\alpha}OHP,\;P_4\;and\;HCD(P<0.05)$. The highest CVBD was observed when $17\alpha20{\beta}OHP$ and HCG were supplemented to media. When oocytes were cultured for 16 hours in media containing $10\~1,000\;ng/ml\;17\alpha20{\beta}OHP,\;17{\alpha}OHP\;and\;P_4$, the rate of GVBD in oocytes cultured in the medium supplemented with 100 ng/ml $17\alpha20{\beta}OHP(65\%)$ was significantly higher than that with $17{\alpha}OHP\;(40\%)\;and\;P_4(35\%)$. The efforts of $17\alpha20{\beta}OHP$ and HCG on GVBD were assessed by various concentration of these hormones. When oocytes were cultured for 60 hours in various media containing $1\~1,000\;ng/ml\;17{\alpha}20{\beta}OHP\;or\;5\~1,000\;IU/ml$ HCG, the GVBD of oocytes was significantly increased in the medium with $10\~100\;ng/ml\;17\alpha20{\beta}OHP$ and 500 IU/ml HCT. When oocytes were cultured in the various media supplemented with $1\~1,000\;ng/ml\;17\alpha20{\beta}OHP\;or\;5\~1,000\;IU/ml$ HCG for 60 hours, the media with $1\~100\;ng/ml\;17\alpha20{\beta}OHP\;or\;50\~1,000IU/ml$ HCG significantly increased in the rate of ovulation. However supplementation with $1,000\;ng/ml\;17\alpha20{\beta}OHP$or 5 IU/ml HCG did not improve the rate of ovulation compared to controls. This results indicate that supplementation of steroid and HCG except $E_2$ can improve the in vitro maturation and ovulation of oocyte in P. fulvidrac; HCG and $17\alpha20{\beta}OHP$ may be more effective than other steroids on oocyte maturation and ovulation in P. fulvidraco.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.6
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• pp.638-642
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• 2001

To investigate the production of $C_{21}$-steroids during the spawning period of spotted sea bass, Lateolabrax maculatus, we have incubated maturing and ovulating follicles with radiolabeled pregnenolone and $17\alpha$-hydroxyprogesterone for 24 hours. The resulting metabolites were analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC), When maturing follicles ($700\sim800{\mu}m$ in diameters) were incubated with radiolabeled precursors, $C_{21}$-metabolites were corticosteroids and $17\alpha$-hydroxy, $20\beta$-dihydroprogesterone ($17\alpha20\beta OHP$). When ovulation follicles ($1,000\sim1,150{\mu}m$ in diameters) were incubated with radiolabeled precursors, the major $C_{21}$-metabolites were $17\alpha20\beta OHP$, $17\alpha$,$20\beta$, 21-trihydroxy-4-pregnen-3-one ($17\alpha20\beta21P$), and corticosterone. Additional chromatography by TLC and HPLC confirmed the presence of radioactive $17\alpha20\beta OHP$ in the maturing follicles, and $17\alpha20\beta OHP$,$17\alpha20\beta21P$ and corticosterone in ovulating follicles. Although $17\alpha20\beta OHP$ was found in a small peak, the synthesis of this steroid suggests that it may play a role in regulating the oocyte maturation process. Whereas ovulation is regulated by both $17\alpha20\beta OHP$ and $17\alpha20\beta21P$ in the spotted sea bass. In addition, an unusual finding was the biosynthesis of corticosterone. Whether this production is responsible for the ovulation, and is an area requiring continued research.

• Fisheries and Aquatic Sciences
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• v.4 no.2
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• pp.70-74
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• 2001

To examine the production of steroids with potential oocyte maturation-inducing activity in the spotted flounder, Verasper variegatus, we have incubated post-vitellogenic oocytes (0.82­0.95mm in diameters) with radiolabeled pregnenolone and $17\alpha-hydroxyprogesterone$. The resulting metabolites were analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). The two main metabolites (progestogens) found in both incubations co-migrated with $17\alpha-hydroxy$, $20\alpha-dihydroprogesterone$ $(17\alpha, 20\alpha OHP)$ and $17\alpha-hydroxy,\;$20\beta-dihydroprogesterone$ (17 a20{30HP). Additional chromatography by HPLC and TLC confirmed the presence of radioactive $17\alpha, 20\alpha OHP$ and a large amount of unknown metabolite. The present study did not reveal in vitro formation of $l7\alpha 20\beta OHP$. Although 1$l7\alpha 20\beta OHP$ was found in a small amount, the synthesis of this steroid suggests that it may play a role in regulating the oocyte maturation process in the spotted flounder.

• Development and Reproduction
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• v.12 no.1
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• pp.107-112
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• 2008

To investigate the $C_{21}$-steroids produced from maturating oocytes in the longchin goby, Chasmichthys dolichognathus, the oocytes ($0.74{\sim}0.97\;mm$) were incubated with radiolabeled $17{\alpha}$-hydroxyprogesterone ($^3H-17{\alpha}OHP$) for 24 hours. The resulting metabolites were analyzed by thin layer chromatography and identified by gas chromatography-mass spectrometry. Two $C_{21}$-steroids, $17{\alpha}$-hydroxy, $20{\alpha}$-dihydroprogesterone ($17{\alpha}20{\alpha}P$) and $17{\alpha}$-hydroxy, $20{\beta}$-dihydroprogesterone ($17{\alpha}20{\beta}P$), were converted from $^3H-17{\alpha}OHP$ in the maturing oocytes. These two main metabolites were detected at 0.80 mm diameter oocytes or greater. In addition, the effects of these metabolites on in vitro germinal vesicle breakdown (GVBD) were tested. The sensitivity of oocytes to the induction of GVBD was greater at $17{\alpha}20{\beta}P$ than $17{\alpha}20{\alpha}P$. This result showed that $17{\alpha}20{\beta}P$ is a major maturation inducing steroid (MIS) in longchin goby, suggesting $17{\alpha}20{\alpha}P$ may play a role in regulating the oocyte maturation process.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.37 no.3
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• pp.192-196
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• 2004

The in vitro effect of bisphenol A (BPA) on ovarian steroidogenesis of the longchin goby (Chasmichthys dolichognathus) was investigated. Oocytes taken during the maturing phase (vitellogenic, fully vitellogenic or germinal vesicle breakdown stage) were incubated with BPA (100 ng/mL) in the presence of exogenous precursor $^{3}H-17\alpha\;hydroxyprogesterone\;(^{3}H-17\alphaOHP).$ Steroids were extracted from the media and the isolated oocytes, and the extracts were separated and identified by thin layer chromatography and gas chromatography-mass spectrometry. The identities of the major metabolites were progestogens $[17{\alpha}-hydroxy,20{\alpha}-dihydroprogesterone\;(17{\alpha}20{\alpha}OHP)\;and\;17{\alpha}-hydrxy,20{\beta}-dihydroprogesterone\;(17{\alpha}20{\beta}OHP),$ androgens [androstenedione (A4) and testosterone (T)] and estrogens [estrone $(E_1)\;and\;estradiol-17{\beta}(E_2)].$ BPA treatment inhibited production of estrogens in all the maturing phases and progestogens in the germinal vesicle migrating stage. Percentage yield of estrogens was decreased with increased yield of androgens. In conclusion, BPA had an inhibitory effect on the conversion of $^3H-17\alphaOHP$ to estrogens and progestogens. These results demonstrate that BPA can act either estrogenic or anti-estrogenic effects.

• Development and Reproduction
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• v.17 no.4
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• pp.421-426
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• 2013

In this study, oocyte steroidogenesis are investigated in relation to oocyte development in the starry flounder, Platichthys stellatus, a marine multiple spawner. Vitellogenic (0.52 and 0.55 mm oocyte diameter) and mature oocytes (0.63, 0.66 and 0.71 mm oocyte diameter) were incubated in vitro in the presence of $[^3H]17{\alpha}$-hydroxyprogesterone ($[^3H]17{\alpha}$-OHP) as a precursor. Steroid metabolites were extracted from the incubated media and oocytes, the extracts were separated and identified by thin-layer chromatography (TLC), high performance liquid chromatography (HPLC) and gas chromatographymass spectrometry (GC-MS). The major metabolites produced from $[^3H]17{\alpha}$-OHP were androgens [androstenedione ($A_4$) and testosterone (T)] and estrogens [$17{\beta}$-estradiol ($E_2$) and estrone ($E_1$)] and progestins [$17{\alpha},20{\alpha}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\alpha}P$) and $17{\alpha},20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$)] in vitellogenic and mature oocytes. The results from this study suggest the potential roles of $E_1$ in the oocytes with diameter 0.52-0.71 mm, $17{\alpha}20{\alpha}P$ and $17{\alpha}20{\beta}P$ at the oocytes of 0.63, 0.66 and 0.71 mm.

• Development and Reproduction
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• v.14 no.2
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• pp.115-121
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• 2010

The in vitro effects of nonylphenol (NP) and 2,2',4,6,6'-pentachlorobiphenyl (PCB104) on ovarian steroidogenesis of the ribbed gunnel, Dictyosoma burgeri were investigated. Oocytes taken during maturation stage were incubated with 100 ng/$m{\ell}$ of NP and PCB104 in the presence of exogenous precursor, $[^3H]-17{\alpha}$-hydroxyprogesterone ($[^3H]-17{\alpha}OHP$). Steroids were extracted from the media and the isolated oocytes, and the extracts were separated and identified by thin layer chromatography. The identities of the major metabolites were testosterone (T) and estradiol-$17{\beta}$ (E2). NP treatment inhibited production of E2 metabolite in the oocytes of 1.2, 1.3 and 1.4 mm although NP inhibited production of T metabolite at the oocytes of 1.1, 1.3 and 1.4 mm. PCB104 treatment inhibited production of T metabolite in the oocytes of all groups and E2 metabolite in the oocytes of 1.2, 1.3 and 1.4 mm. In conclusion, these results suggested that NP and PCB104 had an inhibitory effects on conversion of $[^3H]-17{\alpha}OHP$ to T and E2 during the oocyte maturation process of ribbed gunnel.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.4
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• pp.483-488
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• 2015

We studied oocyte steroidogenesis in the blackfin flounder Glyptocephalus stelleri as a region-specific species, in the East Sea of Korea during the spawning season. Maturing oocytes (0.76, 0.82, 0.88, and 0.91 mm in oocyte diameter) were incubated in vitro in the presence of [$^3H$] $17{\alpha}$-hydroxyprogesterone ($[^3H]17{\alpha}$-OHP) as a precursor. Steroid metabolites were extracted from the incubated medium and oocytes, and the extracts were separated and identified by thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC) and gas chromatographymass spectrometry (GC/MS). The major metabolites produced from $[^3H]17{\alpha}$-OHP were androgens [androstenedione (A4) and testosterone (T)] and estrogens [$17{\beta}$-estradiol (E2) and estrone (E1)] and progestins [$17{\alpha},20{\alpha}$-dihydroxy-4-pregen-3-one ($17{\alpha}20{\alpha}P$) and $17{\alpha}20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$)] in maturing oocytes. The metabolic rate of $17{\alpha}20{\beta}$ was elevated (29.04%) in oocytes measuring 0.88 mm (nucleus migration stage following the induction of germinal vesicle breakdown), but was very low in oocytes measuring 0.76, 0.82, and 0.91 mm (0.42, 0.67, and 2.62%, respectively). From these results, we suggest that $17{\alpha}20{\beta}P$ acts as a maturation-inducing steroid in the blackfin flounder.