• 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.

• Clinical and Experimental Reproductive Medicine
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• v.42 no.2
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• pp.72-76
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• 2015

$17{\alpha}$-hydroxylase and 17,20-lyase are enzymes encoded by the CYP17A1 gene and are required for the synthesis of sex steroids and cortisol. In $17{\alpha}$-hydroxylase deficiency, there are low blood levels of estrogens, androgens, and cortisol, and resultant compensatory increases in adrenocorticotrophic hormone that stimulate the production of 11-deoxycorticosterone and corticosterone. In turn, the excessive levels of mineralocorticoids lead to volume expansion and hypertension. Females with $17{\alpha}$-hydroxylase deficiency are characterized by primary amenorrhea and delayed puberty, with accompanying hypertension. Affected males usually have female external genitalia, a blind vagina, and intra-abdominal testes. The treatment of this disorder is centered on glucocorticoid and sex steroid replacement. In patients with $17{\alpha}$-hydroxylase deficiency who are being raised as females, estrogen should be supplemented, while genetically female patients with a uterus should also receive progesterone supplementation. Here, we report a case of a 21-year-old female with $17{\alpha}$-hydroxylase deficiency who had received inadequate treatment for a prolonged period of time. We also include a brief review of the recent literature on this disorder.

• Journal of Life Science
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• v.23 no.10
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• pp.1230-1238
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• 2013

The regulatory effect of the tumor-suppressor protein p53 on the apoptogenic activity of $17{\alpha}$-estradiol ($17{\alpha}-E_2$) was compared between HCT116 ($p53^{+/+}$) and HCT116 ($p53^{-/-}$) cells. When the HCT116 ($p53^{+/+}$) and HCT116 ($p53^{-/-}$) cells were treated with $2.5{\sim}10{\mu}M$ $17{\alpha}-E_2$ for 48 h or with $10{\mu}M$for various time periods, cytotoxicity and an apoptotic sub-$G_1$ peak were induced in the HCT116 ($p53^{+/+}$) cells in a dose- and time-dependent manner. However, the HCT116 ($p53^{-/-}$) cells were much less sensitive to the apoptotic effect of $17{\alpha}-E_2$. Although $17{\alpha}-E_2$ induced aberrant mitotic spindle organization and incomplete chromosome congregation at the equatorial plate, $G_2/M$ arrest was induced to a similar extent in both cell types. In addition, $17{\alpha}-E_2$-induced activation of Bak and Bax, ${\Delta}{\Psi}m$ loss, and PARP degradation were more dominant in the HCT116 ($p53^{+/+}$) than in the HCT116 ($p53^{-/-}$) cells. In accordance with enhancement of p53 phosphorylation (Ser-15) and p53 levels, p21 and Bax levels were elevated in the HCT116 ($p53^{+/+}$) cells treated with $17{\alpha}-E_2$. The HCT116 ($p53^{-/-}$) cells exhibited barely or undetectable levels of p21 and Bax, regardless of $17{\alpha}-E_2$ treatment. On the other hand, although the level of Bcl-2 was slightly lower in the HCT116 ($p53^{+/+}$) than in the HCT116 ($p53^{-/-}$) cells, it remained relatively constant after the $17{\alpha}-E_2$ treatment. Together, these results show that among the components of the $17{\alpha}-E_2$-induced apoptotic-signaling pathway, which proceeds through mitotic spindle defects causing mitotic arrest, subsequent activation of Bak and Bax and the mitochondria-dependent caspase cascade, leading to PARP degradation, $17{\alpha}-E_2$-induced activation of Bak and Bax is the upstream target of proapoptotic action of p53.

• 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.

• Development and Reproduction
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• v.23 no.2
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• pp.111-117
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• 2019

This study focused on the association of prostaglandins and a progestin, $17{\alpha}$, $20{\beta}P$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$) during the ovulation process in longchin goby, Chasmichthys dolichognathus. We performed several in vitro experiments using $850-920{\mu}m$ diameter oocytes which were at the migratory nucleus stage. With the $890-920{\mu}m$ diameter oocytes, no significant difference in ovulation was observed in any of the prostaglandins (PGE1, PGE2, and $PGF2{\alpha}$) treated groups although PGE2 and $PGF2{\alpha}$ at concentrations of 50 ng/mL increased ovulation slightly compared with controls; however, $17{\alpha}20{\beta}P$ production was stimulated with PGE1 alone at low concentrations (5 ng/mL). In $850{\mu}m$ diameter oocytes, $PGF2{\alpha}$ at concentrations of 50 and 500 ng/mL resulted in a significant increase in ovulation. $17{\alpha}20{\beta}P$ (50 ng/mL) alone had no observable effect on ovulation, but in the combined of $PGF2{\alpha}$ 50 or 500 ng/mL it caused the greatest effect on ovulation. The sensitivity of oocytes to the induction of ovulation varies between 850 and $890-920{\mu}m$, it appeared to vary depending on the migration status of nucleus. These results suggest that $PGF2{\alpha}$ (or combined of $17{\alpha}20{\beta}P$) was more potent in inducing ovulation of the longchin goby.

• Development and Reproduction
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• v.10 no.2
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• pp.127-133
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• 2006

[ $17\;{\alpha}-hydroxylase/17,20-lyase(P450_{C17})$ ] is the key enzyme mediating the conversion of progesterone to $17\;{\alpha}-hydroxyprogesterone$, ultimately to androstenedione during steroidogenesis. R. dybowskii's ovarian $P450_{C17}$ cDNA was cloned to understand the regulatory mechanism of ovarian steroidogenic pathway at the molecular level in amphibian. A 2.5kb cDNA clone encoding a single open-reading frame with a 519 deduced amino acid was isolated with the screening of ovarian cDNA library. This sequence contained the three highly conserved domains as seen in $P450_{C17}$ of other species. The comparison of amino acid sequence of Rana $P450_{C17}$ with other animal's $P450_{C17}$ showed relatively high identity with 76% in Xenopus, 63% in chicken, 60% in rainbow trout, and 45% in human. Phylogenic analysis also indicated that Rana $P450_{C17}$ gene was evolutionary well conserved among vertebrate. Northern analysis indicated that the two different sizes of $P450_{C17}$ transcripts with approximately 2.5 and 3.6kb were detected in ovary tissue, but not in other tissues. The expression vector of Rana $P450_{C17}$ clearly showed the $17\;{\alpha}-hydroxylase$ activity converting the exogenous progesterone into $17\;{\alpha}-hydroxyprogesterone$ in the nonsteroidogenic COS-1 cells. Therefore, Rana $P450_{C17}$ cDNA is very useful to investigate the molecular mechanism of the ovarian steroidogenesis in amphibian.

• Biomedical Science Letters
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• v.14 no.3
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• pp.131-137
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• 2008

Adipogenesis is a complex sequence of events that culminates in the differentiation of fibroblast-like preadipocytes into specialized lipid-filled adipocytes and also involves a cascade of expression of many transcription factors such as peroxisome proliferator-activated receptor ${\gamma}(PPAR{\gamma})$ and CCAAT/enhancer-binding proteins (C/EBPs). $PPAR{\gamma}$ and C/EBPs transcriptionally transactivate adipocyte specific genes, including fatty acid transport protein (FAT/CD36) and leptin. To determine whether $17{\beta}$-estradiol modulates $C/EBP{\alpha}$ actions on adipogenesis in high fat diet-fed female ovariectomized (OVX) C57BL/6 mice, mice were treated with $17{\beta}$-estradiol for 7 days and the effects of $17{\beta}$-estradiol on adipose tissue mass and expression of adipocyte specific gene as well as $C/EBP{\alpha}$ were measured. Compared to vehicle-treated OVX control mice, OVX mice treated with $17{\beta}$-estradiol for 7 days had lower adipose tissue weights that were similar to weights in high fat diet-fed sham-operated (Sham) mice. OVX mice showed the increased expression of $C/EBP{\alpha}$ mRNA compared with Sham mice. However, $17{\beta}$-estradiol treatment in OVX mice inhibited OVX induced-$C/EBP{\alpha}$ activation, indicating that $17{\beta}$-estradiol may act as an inhibitor of $C/EBP{\alpha}$ action. Moreover, $17{\beta}$-estradiol decreased mRNA levels of adipocyte marker genes, such as lipoprotein lipase, FAT/CD36 and leptin, to levels in Sham mice. These results suggest that down-regulation of adipogenesis by $17{\beta}$-estradiol may be due to reduced adipose $C/EBP{\alpha}$ activities in female OVX C57BL/6 mice.

• 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.

• Bulletin of the Korean Chemical Society
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• v.30 no.10
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• pp.2398-2402
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• 2009

Ten steroidal compounds as impurities in canrenone were isolated from the enriched mother liquor by using various chromatographic methods. Their structures were elucidated by spectrometric analysis, among which three new compounds were characterized as 3-(3-oxo-7$\alpha$-(ethoxycarbonyl)methyl-17$\beta$-hydroxy-4-androsten-17$\alpha$-yl) propionic acid $\gamma$-lactone (1), 3-(3-oxo-7$\alpha$-ethoxy-17$\beta$-hydroxy-4-androsten-17$\alpha$-yl) propionic acid $\gamma$-lactone (2) and 3-(3-oxo- 5$\beta$-propionic acid-$\gamma$-lactone-6$\beta$,17$\beta$-hydroxy-4-androstan-17$\alpha$-yl) propionic acid $\gamma$-lactone (3).

• Korean Journal of Food Science and Technology
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• v.30 no.2
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• pp.434-438
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• 1998

The antioxidant effects of 700 ppm ginseng extract, 100ppm caffeic acid, ferulic acid, vanillic acid, or ${\alpha}-topherol$ on the 1% linoleic acid aqueous buffer system was studied by measuring malondialdehyde (MDA) and headspace oxygen. The compounds showed antioxidant activities in the following order: $caffeic\;acid{\geq}ferulic\;acid\;>\;{\alpha}-tocopherol$>ginseng extract>vanillic acid, with the oxidation inhibition ratio of 63.5, 62.9, 52.3, 51.2 and 5.6% of the control according to MDA results. The mixture of 100 ppm caffeic acid, 100 ppm ${\alpha}-tocopherol$ and 700 ppm ginseng extract had a high oxidation inhibition ratio of 91.2%. Headspace oxygen results had a similar trend with the MDA results. Headspace oxygen results showed that the antioxidant activities were in order of ferulic acid>caffeic acid>${\alpha}-tocopherol$>ginseng extract>vanillic acid and headspace oxygen contents were 18.56, 17.78, 17.17, 16.65 and 15.95%, respectively.