• Title/Summary/Keyword: Sex hormones

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Establishment of a library of fragments for the rapid and reliable determination of anabolic steroids by liquid chromatography-quadrupole time of flight-mass spectrometry

  • Do, Jung-Ah;Noh, Eunyoung;Yoon, Soon-Byung;Choi, Hojune;Baek, Sun-Young;Park, Sung-Kwan;Lee, Sang-Gyeong
    • Analytical Science and Technology
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    • v.30 no.1
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    • pp.10-19
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    • 2017
  • Anabolic steroids have similar structures to testosterone, both of which promote the growth of muscle mass and increase strength. However, the side effects of anabolic steroid use may lead to heart attacks or strokes. Additionally, the excessive use of steroids inhibits the production of the sex hormones in the body via a negative feedback loop, which results in testicular atrophy in males and amenorrhea in females. Currently, the method of choice used to test for the presence of anabolic steroids is GC-MS. However, GC-MS methods require chemical derivatization of the steroid sample to ensure compatibility with the analytical method; therefore, analysis of many different samples is difficult and time consuming. Unlike GC-MS, the liquid chromatography-quadrupole-time of flight mass spectrometry (LC-Q-TOF-MS) method is suitable for many samples. Twenty-two different anabolic steroids were analyzed by LC-Q-TOF-MS with various collision energies (CE). Accurate mass spectral data were obtained using a Q-TOF-MS equipped with an electro-spray ionization source and operated in the positive MS/MS mode for several classes of steroids that are often the targets of testing. Based on the collected data, fragmentation pathways were carefully elucidated. The high selectivity and sensitivity of the LC-Q-TOF-MS instrument combined with these fragmentation pathways offers a new approach for the rapid and accurate screening of anabolic steroids. The obtained data from the 22 different anabolic steroids will be shared with the scientific community in order to establish a library to aid in the screening of illegal anabolic steroids.

Effects of Steroid Hormones on $In$ $Vitro$ GVBD and Oocyte Steroidogenesis in Blacktip Grouper, $Epinephelus$ $fasciatus$

  • Hwang, In-Joon;Kim, Seol-Ki;Choi, Sang-Jun;Lee, Chi-Hoon;Lee, Young-Don;Kim, Hyung-Bae;Baek, Hea-Ja
    • Development and Reproduction
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    • v.16 no.1
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    • pp.39-45
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    • 2012
  • To verify the sex steroids which are involved in oocyte maturation of the blacktip grouper, $Epinephelus$ $fasciatus$, we incubated vitellogenic oocytes (0.41 and 0.50 mm in average diameter) in the presence of exogenous steroid precursor ($[^3H]17{\alpha}$-hydroxyprogesterone). Steroids were extracted, separated and identified by thin layer chromatography. The major metabolites produced were androstenedione, estradiol-$17{\beta}$, estrone and progestogens. Progestogen metabolites in the oocytes of 0.50 mm were more abundant than those of 0.41 mm. Also, we investigated the $in$ $vitro$ effects of human chorionic gonadotropin (HCG; 5, 50 and 500 $IU/m{\ell}$), $17{\alpha},20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$) and $17{\alpha},20{\beta}$-trihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}21P$; 5, 50 and 500 $ng/m{\ell}$, respectively) on oocyte maturation. In the oocytes of 0.41 mm, treatment with 50 IU HCG stimulated GVBD ($55.30{\pm}1.20%$) compared with controls ($32.41{\pm}3.13%$, $p$<0.05). In the oocytes of 0.50 mm, treatment of $17{\alpha}20{\beta}P$ (50 and 500 $ng/m{\ell}$) stimulated GVBD ($50.13{\pm}2.52$ and $51.77{\pm}5.91%$, respectively) compared with controls ($36.81{\pm}2.89%$, $p$<0.05). Treatment with 500 IU HCG also stimulated GVBD ($49.59{\pm}5.15%$) compared with controls ($p$<0.05). Taken together, these results suggested that both HCG and $17{\alpha}20{\beta}P$ were effective on in vitro oocyte maturation and $17{\alpha}20{\beta}P$ may act as a maturation inducing hormone in blacktip grouper.

The Effect of Interferon-${\alpha}$ and bFGF on the Proliferation of Cultured Leiomyoma and Myometrial Cells (자궁근종과 자궁평활근 세포분열에 있어 Interferon-${\alpha}$ 및 basic Fibroblast Growth Factor (bFGF)의 효과)

  • Lee, B.S.;Park, J.S.;Kim, J.Y.;Bae, S.W.;Park, K.H.;Cho, D.J.;Lee, K.d;Kim, J.W.;Song, C.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.3
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    • pp.355-359
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    • 1997
  • Leiomyomas, which are the commonest pelvic tumors in women, are originated from myometrial cells. Although the exact initial pathophysiologic event of the leiomyoma is not known, recent evidences suggested that the effects of sex steroid hormones in the process of tumor growth are mediated by local production of growth factors including epidermal growth factor (EGF), insulin-like growth factor-I (IGF-I) and insulin-like growth factor-II (IGF-II). If we look at the effects of other cytokines, it was suggested that basic fibroblast growth factor (bFGF) may stimulate the proliferation of myometrial and leiomyomas cells. And it was reported that interferon-${\alpha}$ inhibit the action of bFGF. Therefore, we examined the effect of bFGF and interferon-${\alpha}$ on the proliferation of leiomyoma and myometrial cells. bFGF stimulated the myometrial and leiomyoma cells significantly at the concentration of 1ng/ml (p<0.05) and 5ng/ml (p<0.05). However, Interferon-${\alpha}$ inhibited the cell proliferation of myometrial and leiomyoma cells significantly at the concentration of 100U/ml (p<0.05) and 1000U/ml (p<0.05). And the stimulated effects of bFGF with the various concentration on the myometrial and leiomyoma cells ware inhibited by interferon-${\alpha}$ with 100U/ml. Therefore, we concluded that bFGF may stimulate the myometrial and leiomyoma cell proliferation and interferon-${\alpha}$ may inhibit the myometrial and leiomyoma cell proliferation through blocking the effect of basic fibroblast growth factor.

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Amylase Release from Pancreatic Slices of Rat Treated with Adrenergic Drugs (아드레나린성 약물 전처치 흰쥐의 취절편 효소분비에 관한 실험)

  • Kim, Kyung-Hwan;Kim, Hea-Young;Ahn, Young-Soo;Lee, Woo-Choo;Hong, Sa-Suk
    • The Korean Journal of Pharmacology
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    • v.20 no.2
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    • pp.49-57
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    • 1984
  • The exocrine pancreatic secretion is controlled mainly by gastrointestinal hormones as well as cholinergic nerves. The adrenergic influence on exocrine pancreas is thought not to he important and the evidences supporting this contention are still contradictory. In an effort to elucidate the adrenergic influence on the exocrine pancreas, we have determined the amylase release from pancreatic slices of rats treated with adrenergic drugs. The albino rats of either sex, weighing $60{\sim}80\;g$, were decapitated and the uncinate pancreata were isolated and incubated in screw top vials containing 2 ml krebs-Ringer bicarbonate buffer solution gassed with 95% $O_2$ and 5% $CO_2$. These vials were shaken continuously in a waterbath maintained at $37^{circ}C$, and enzyme release was stimulated with acetylcholine$(10^{-5}M)$. For chronic treatment methoxamine$(an\;{\alpha}-adrenergic\;agonist,\;5\;mg/kg)$, isoproterenol (a\;{\beta}-adrenergic\;agonist,\;10\;mg/kg) and reserpine (0.5 mg/kg) along with cholecystokinin octapeptide$(CCK-op,\;2{\mu}g/kg)$ were given i.p. in rats daily for 3, 5, 7, 9 or 12 days. For acute experiment these drugs were added directly to the incubation medium in a concentration of $10^{-5}M$ except CCK-OP $(10^{-9}M)$. The results are summarized as follows. 1) The addition of methoxamine, isoproterenol or reserpine to the incubation medium containing pancreatic slices augmented the release of amylase induced by acetylcholine and among them the effect of isoproterenol was most prominent. 2) Chronic treatment of methoxamine or reserpine caused enhancement of acetylcholine response in amylase release from pancreatic slice throughout the experimental period, but the amylase release was less than that of control by 12 days isoproterenol treatment. 3) In the pancreatic slices obtained from 12 days treatment of CCK-OP, the amylae release responding to acetylcholine was enhanced. By these finding it is suggested that methoxamine, isoproterenol and reserpine had marked influence on the exocrine pancreatic functions in rats and that these effects are due to their inherent actions rather than sympathetic nerve or adrenergic receptor function.

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Understanding of Intrauterine Environment Changes based on Proteomics and Bioinformatics during Estrous Cycle (단백체학과 생물정보학을 이용한 자궁 내 환경의 이해)

  • Lee, Sang-Hee;Lee, Seunghyung
    • Journal of Life Science
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    • v.29 no.5
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    • pp.621-630
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    • 2019
  • Fertilization is the beginning of a new life that occurs in the female uterine. The female reproductive tract is composed ovary, oviduct, uterine, vagina and cervix, their physiological features are regulated by estrous cycle. Of these, uterine is a main point to establish embryo development and implantation, and intercommunication between embryo and uterine environment is necessary for suitable pregnancy. Endometrium is part of the uterine, its morphology is repetitively changed by hormones, and characteristic of uterine fluid from endometrium is also changed. Recently, massive proteins of endometrium and uterine fluid can be detected according to develop proteomics and bioinformatics and have been accelerated the understanding of the reproductive biology fields. Moreover, the massive protein information is actively studying with deeply studied theory such as sex hormone signal pathway and angiogenesis in mammals. In this paper, we review understanding of endometrium remodeling, uterine gland and fluid during estrous cycle, additionally studies on endometrium and uterine fluid based on proteomics techniques. Lastly, we introduced methods of the protein-protein correlation using bioinformatics tool that interaction with hormone receptors, representative angiogenetic factors and detected proteins using proteomics in endometrium and uterine fluid. This review will be useful to understanding the study on search of new cell mechanism in endometrium and uterine fluid.

Caspase-3 Activation is Associated with Granulosa Cell Apoptosis during Follicular Atresia in Porcine Ovary (돼지 폐쇄난포내 과립세포의 자연세포사 시 캐스파제-3의 활성화)

  • Kim, Jong-Min;Chung, Jin-Yong;Kim, Ji-Young;Oh, Seung-Hoon;Song, Kang-Won;Do, Byoung-Rok;Kim, Sang-Soo;Jung, Jin;Lee, Chang-Joo;Yoon, Yong-Dal
    • Development and Reproduction
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    • v.10 no.1
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    • pp.1-7
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    • 2006
  • Ovarian follicular atresia in mammals is finely regulated by gonadotropins and sex steroid hormones. It is well known that granulosa cell pyknosis is a common cytological feature of atretic follicles in the ovary. The present study hypothesized that granulosa cell pyknosis during follicular atresia might be related to apoptotic process and associated with caspase-3 activation. Healthy (normal) and atretic follicles were isolated from porcine ovaries based on macro-morphological criteria. Isolated follicles were either processed for histological observation or used for collection of granulosa cells by aspiration. Hoechst 33258 staining of the cells showed a significantly higher number of fragmented nuclei, a typical morphological feature of apoptotic cell, in granulosa cells from atretic follicles than those from healthy follicles. In addition, the rate of cell death was significantly higher in granulosa cells from atretic follicles than healthy follicles, as measured by flow-cytometric cell cycle analysis. In situ detection of apoptotic cells by TUNEL revealed that apoptosis was mostly restricted to granulosa cells in follicles. Theca cells were TUNEL-negative. Finally, it has been shown by caspase-3 activity assay that granulosa cells from atretic follicles retain a higher caspase-3 activity compared to healthy follicles. Taken together, it is suggested that granulosa cell degeneration during folliclar atresia occurs by caspase-3-dependent apoptotic fashion.

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Studies on the Sex Hormone Level is Serum of Puerperal Sow (돼지에 있어서 분만전후(分娩前後)의 혈청(血淸) 성(性) Hormone 수준(水準)에 관(關)한 연구(硏究))

  • Lee, Kyu Seung;Park, Chang Sik
    • Korean Journal of Agricultural Science
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    • v.8 no.1
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    • pp.64-71
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    • 1981
  • The present study was carried out to study the serum concentration of peptide and steroid hormones in puerperal sow. Eight crossbred sows were used for collection of blood samples from day 20 prepartum to day 20 postpartum. FSH, LH, prolactin, estradiol-$17{\beta}$, progesterone and cortisol were assayed by radioimmunoassay methods. The mean serum FSH did not vary during the puerperal period and ranged from $8.1{\pm}1.8mIU/ml$ to $9.0{\pm}2.3mIU/ml$. LH concentrations increased from $2.6{\pm}0.3mIU/ml$ at day 20 prepartum to $3.9{\pm}1.1mIU/ml$ at the time of parturition, reached $3.2{\pm}0.9mIU/ml$ by day+2 and remained quite constant therafter. Prolactin reached a peak mean level of $68.5{\pm}9.5ng/ml$ at day 0. Estradiol-$17{\beta}$ increased from $205.0{\pm}29.5pg/ml$ at day 6 prepartum to $425.0{\pm}35.0pg/ml$ at the time of parturition. Progesterone remained fairly constant ($18.4{\pm}1.6$ to $20.2{\pm}2.1ng/ml$) from 20 to 6 days before parturition, began to decline on day-2, reached $0.9{\pm}0.3ng/ml$ by day+2 and remained quite constant thereafter. Cortisol reached a peak level of $86.5{\pm}10.5ng/ml$ at the day 0.

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Effect of Growth Hormone and Androgen on Vitellogenin and Estrogen Receptor Gene Expression in the Japanese eel, Anguilla japonica (뱀장어 Vitellogenin과 Estrogen 수용체 유전자 발현에 대한 성장호르몬 및 웅성호르몬의 영향)

  • Kwon, Hyuk-Chu;Choi, Seong-Hee;Kim, Eun-Hee;Kwon, Joon-Yeong
    • Development and Reproduction
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    • v.10 no.2
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    • pp.97-103
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    • 2006
  • Vitellogenin(Vg) is a sex specific serum protein present in sexually maturing female blood of oviparous vertebrates. Estrogen($E_2$) is a main inducer of hepatic Vg synthesis. We investigated the effects of androgen and growth hormone(GH) on regulation of Vg and estrogen receptor(ER) genes in Japanese eel. Immature eels($200{\sim}250\;g$) were given a single injection of $E_2(5{\sim}5,000\;{\mu}g/kg\;bw)$ alone, or in combination with eel recombinant GH(eGH, $1{\sim}10\;{\mu}g/kg$) or methyltestosterone(MT, $1{\sim}5\;mg/kg$) and sacrificed 10 days after the hormone treatments. Expression levels of ER and Vg genes from the liver were determined by means of reverse transcription and polymerase chain reaction(RT-PCR). Administration of $E_2$ stimulated Vg gene expression in a dose dependent manner. Levels of Vg mRNA after the injection of $E_2(500\;{\mu}g/kg)$ with MT(5mg/kg) or eGH($10\;{\mu}g/kg$) were much higher than in that of $E_2$ alone($500\;{\mu}g/kg$). Whereas, injection of either vehicle, eGH ($10\;{\mu}g/kg$) or MT(5mg/kg) alone did not induce the expression of Vg gene in the liver. ER mRNA was detected from the fish treated with vehicle alone. $E_2$ injection($5{\sim}500\;{\mu}g/kg\;bw$) increased this ER expression but dose dependent response was not clear. Addition of MT(5mg/kg) or eGH($10\;{\mu}g/kg$) did not affect $E_2-stimulated$ ER mRNA expression. This study confirms the necessity of $E_2$ as the primary factor for Vg gene expression and requirement of additional hormones such as MT or GH for the full expression of Vg mRNA, and suggests that the additive effect of MT or GH on Vg gene expression would be mediated by some unknown factors other than ER.

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Premenstrual Syndrome and its Relationship with Previous Abuse History and Dissociative Symptoms (월경전기 증후군과 해리증상 및 과거 학대 경험과의 연관성)

  • Kim, Jong-In;Choi, Tae-Kyou;Yoon, Jae-Hang;Suh, Shin-Young
    • Korean Journal of Psychosomatic Medicine
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    • v.16 no.2
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    • pp.112-119
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    • 2008
  • Objectives : Premenstrual syndrome(PMS) is known to occur in more than 80% of reproductive women. Since PMS is closely related to changes in sex hormones, biological factors are suspected to be associated with the syndrome, but there have been no consistent reports regarding biological causes. In that sense, it is postulated that the occurrence of PMS is much dependent on the biological aspects, but that the severity or characteristics of the syndrome is more dependent on the psychological factors. Studies focusing on psychological factors are based on the theory that symptoms of PMS and previous psychologically traumatic events are closely related. If the theory is valid, it can be further postulated that symptom severity of PMS is associated with dissociative symptoms, since traumatic events are known to be related to dissociative symptoms. In this study, we tried to find out the association of PMS symptom severity with previous abuse history and dissociative symptoms. Methods : Subjects for the study were 377 nurses working in Seoul, Kyung-gi, and Kyung-book. 183 subjects who submitted valid data entered the study. The presence and the symptom severity of PMS were rated using Daily Records of Severity of Problems(DRSP) and Shortened Premenstrual Assessment Form(SPAF). Dissociative symptoms were evaluated with Dissociation Experience Scale Korean version(DES-K). Previous physical/verbal abuse, sexual abuse, and parental spouse abuse experience were rated with correspondent abuse scales. Results : Subjects were divided into 3 groups as No PMS group, mild to moderate PMS group, and severe PMS group according to SPAF total score. There was a statistically significant difference in DES-K total score among 3 groups($x^2=14.966$, df=2, p=0.001). Physical/verbal abuse scale($x^2=14.397$, df=2, p=0.001), sexual abuse scale($x^2=8.376$, df=2, p=0.015), and parental spouse abuse scale($x^2=9.322$, df=2, p=0.009) also revealed a significant difference among 3 groups. Symptom severity of PMS using SPAF total score showed a positive correlation both with degree of dissociative experience and previous abuse experience. There was a statistically significant difference in both dissociative experience and previous abuse experience among 3 groups. Conclusion : These results show that there can be a possible association among PMS symptoms, dissociative symptoms, and previous psychologically traumatic experience.

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A Study on Phthalate Analysis of Nail Related Products (네일 관련 제품들의 프탈레이트 분석에 관한 연구)

  • Rark, Sin-Hee;Song, Seo-Hyeon;Kim, Hyun-Joo;Cho, Youn-Sik;Kim, Ae-Ran;Kim, Beom-Ho;Hong, Mi-Yeun;Park, Sang-Hyun;Yoon, Mi-Hye
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.3
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    • pp.217-224
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    • 2019
  • Phthalates, endocrine disrupting chemicals, are similar in structure to sex hormones and mainly show reproductive toxicity and developmental toxicity. In this study, we analyzed 11 phthalates, including 3 kinds of phthalates prohibited in cosmetic use and 8 kinds of phthalates regulated in 'Common standards for children's products safety' and EU cosmetic regulation (EC No. 1223/2009). The phthalate analysis was optimized using GC-MS/MS. In analytical method validation, this method was satisfied in specificity, linearity, recovery rate, accuracy and MQL. Therefore, we used this method to analyze 82 products of Nail cosmetics & polish. Although six phthalates such as DBP, BBP, DEHP, DPP, DIBP and DIDP were detected at concentrations of $1.0{\sim}59.8{\mu}g/g$g, they were suitable to Korean cosmetic standards. DIBP and DBP were detected at concentration of $1.1{\sim}2.6{\mu}g/g$ in artificial nail, DBP and DEHP were $1.4{\sim}2.5{\mu}g/g$ in glue for nails, and DIBP, DBP, and DEHP were $2.5{\sim}33.3{\mu}g/g$ in nail stickers. Although substances such as DBP and DEHP in artificial nail, Glue for nails, and nail stickers were detected, they were suitable to 'Common safety standards for children's products. DIBP is not a regulated substance in Korea but showed the third highest detection rate following DBP (84.6%) and DEHP (63.4%). The concentration of phthalates detected in nail products is considered to be safe in current standards but continuous monitoring and research about non-regulated substances are also needed to be considered.