• Title/Summary/Keyword: Estradiol activity

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Production and Characterization of vitellogenin monoclonal antibody on the Scorpion fish Sebastiscus marmoratus (쏨뱅이, Sebastiscus marmoratus의 vitellogenin 단클론 항체생산 및 특성에 관한 연구)

  • Kim, Young-Ju;Lim, Yoon-Kyu;Yeo, In-Kyu
    • Journal of fish pathology
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    • v.26 no.3
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    • pp.241-254
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    • 2013
  • In order to establish bio-marker systems for the screening of endocrine-disrupting chemicals contaminated in various environment, Vitellogenin(Vtg) bio-marker have been developed to detect Scorpion fish's(Sebastiscus marmoratus) Vtg. Vtg has been induced by administration of estradiol into S. marmoratus, and purified by gel filtration and ion-exchange chromatography from serum of the fish. After immunization of the purified Vtg into BALB/c mouse, hybridomas secreting anti-Vtg antibodies have been produced. The size of induced Vtg in the serum was about 440 kDa by gel filtration using Sepharose CL-6B. By SDS-PAGE analysis, the main band of Vtg, however, was at 175 kDa, and several minor bands have been detected with the main band. Eight different monoclonal antibodies have been produced from established hybridomas and the antibodies did not cross-react with sera from different species of fishes tested in this study except with that of Sebastes hubbsi. These results suggested that the monoclonal antibody of S28 and S15 can used as capture and tracer antibodies for ELISA and ICG assays. The detection systems developed in this study can be used as Bio-marker assays to check endocrine disrupting activity of various chemicals as well as to detect known endocrine disrupting chemicals contaminated in environment.

Ginsenoside Rg1 activates ligand-independent estrogenic effects via rapid estrogen receptor signaling pathway

  • Gao, Quan-Gui;Zhou, Li-Ping;Lee, Vien Hoi-Yi;Chan, Hoi-Yi;Man, Cornelia Wing-Yin;Wong, Man-Sau
    • Journal of Ginseng Research
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    • v.43 no.4
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    • pp.527-538
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    • 2019
  • Background: Ginsenoside Rg1 was shown to exert ligand-independent activation of estrogen receptor (ER) via mitogen-activated protein kinase-mediated pathway. Our study aimed to delineate the mechanisms by which Rg1 activates the rapid ER signaling pathways. Methods: ER-positive human breast cancer MCF-7 cells and ER-negative human embryonic kidney HEK293 cells were treated with Rg1 ($10^{-12}M$, $10^{-8}M$), $17{\beta}$-estradiol ($10^{-8}M$), or vehicle. Immunoprecipitation was conducted to investigate the interactions between signaling protein and ER in MCF-7 cells. To determine the roles of these signaling proteins in the actions of Rg1, small interfering RNA or their inhibitors were applied. Results: Rg1 rapidly induced $ER{\alpha}$ translocation to plasma membrane via caveolin-1 and the formation of signaling complex involving linker protein (Shc), insulin-like growth factor-I receptor, modulator of nongenomic activity of ER (MNAR), $ER{\alpha}$, and cellular nonreceptor tyrosine kinase (c-Src) in MCF-7 cells. The induction of extracellular signal-regulated protein kinase and mitogen-activated protein kinase kinase (MEK) phosphorylation in MCF-7 cells by Rg1 was suppressed by cotreatment with small interfering RNA against these signaling proteins. The stimulatory effects of Rg1 on MEK phosphorylation in these cells were suppressed by both PP2 (Src kinase inhibitor) and AG1478 [epidermal growth factor receptor (EGFR) inhibitor]. In addition, Rg1-induced estrogenic activities, EGFR and MEK phosphorylation in MCF-7 cells were abolished by cotreatment with G15 (G protein-coupled estrogen receptor-1 antagonist). The increase in intracellular cyclic AMP accumulation, but not Ca mobilization, in MCF-7 cells by Rg1 could be abolished by G15. Conclusion: Ginsenoside Rg1 exerted estrogenic actions by rapidly inducing the formation of ER containing signalosome in MCF-7 cells. Additionally, Rg1 could activate EGFR and c-Src ER-independently and exert estrogenic effects via rapid activation of membrane-associated ER and G protein-coupled estrogen receptor.

Immunomodulatory Activities by Difference in Molecular Size of the Proteoglycan Extracted from Ganoderma lucidum IY009 (Ganoderma lucium IY009 유래 단백다당류의 분자량 차이에 따른 면역증강활성)

  • Lee, June-Woo;Baek, Seong-Jin;Bang, Kwang-Woong;Kim, Yong-Seuk;Kim, Kwang-Soo;Chun, Uck-Han
    • The Korean Journal of Mycology
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    • v.29 no.1
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    • pp.15-21
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    • 2001
  • This study was conducted to investigate the immunomodulatory activities of proteoglycan extracted from cultured mycelia of Ganoderma lucidum IY009. The proteoglycan contained two polymer peaks, one was the higher MW peak of 2,000 kD and the other was low peaks of 12kD. To understand the part of strong pharmaceutical activity between two peak, the proteoglycan was separated by ultrafiltration and column chromatography and then examined the various pharmaceutical effects. High molecular weight fraction possesing high content of ${\beta}-linked$ glucan was exhibited high antitumor activity, against sarcoma 180 bearing ICR mouse. And also, anticomplementary activity was highly observed in high molecule fraction than low it fraction. When the raw 264.7 and murine peritoneal macrophage treated with low fraction, high fraction and other stimuli. The activities inducing tumor necrosis factor of the high factions were $2.2{\sim}2.5$ times stronger than that of low fraction.

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Effects of Soy Isoflavone on the Bone Metabolism in Ovariectomized Growing Rats (콩 이소플라본이 난소를 절제한 성장기 흰쥐의 골격대사에 미치는 영향)

  • Kim, Young-Kyoung;Lee, Heon-Ok;Om, Ae-Son
    • Korean journal of food and cookery science
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    • v.21 no.5
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    • pp.725-732
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    • 2005
  • This study attempted to investigate if the soy isoflavone, genistein, influences bone metabolism in ovariectomized, 4-week-old female Wister rats. All the rats were divided into sham (SH) and ovariectomized (OVX) groups consisting of OVX-17${\beta}$-estradiol($10\;{\mu}g/kg$ b.w.), OVX-1mg or 5 mg or 10mg of genistein/kg b.w. The rates were allowed ad libitum access to foods and water for 8 weeks. The Results showed that body weight had significantly increased in the OVX group compared to the SH group (p<0.05) and was not different among the OVX-GEN and OVX-ES groups and the OVX group. The liver and uterus weights in the OVX groups were slighter than those in SH group (p<0.05). The kidney weight in the OVX groups was decreased compared to in that in the SH group but was not significantly different among all OVX groups. Femoral length in the OVX groups was longer than in the SH group and was not different. Rats in the OVX groups had higher creatinine levels than those in the SH group and hydroxyproline level did not differ significantly among any of the groups. Serum ALP activity and Ca in bone, feces, urine and serum did not change among the groups. Bone mineral density (BMD) decreased in the OVX groups compared to the SH group and was slightly increased by feeding genistein (p>0.05). Breaking force and stiffness did not change by ovariectomy and feeding genistein. Hence, these results suggested that estrogen may affect bone mineralization in growing rats and that genistein be may involved in the prevention of bone loss. However, more studies are needed to identify the proper mechanism of genistein and bone formation.

Estrogeicity of Genistein and Bisphenol A (콩류식품의 주성분인 Genistein과 식품포장재 및 용기에 사용되는 Bisphenol A의 에스트로젠 효과에 관한 연구)

  • 강경선;이영순;신광순
    • Journal of Food Hygiene and Safety
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    • v.13 no.2
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    • pp.106-111
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    • 1998
  • This study has been focused on both estrogenic and proliferating activity of genistein (GEN) and bisphenol A (BPA). GEN and BPA enhance the proliferation of estrogen-dependent MCF-7 human breast cancer cells at concentrations as low as 100 nM of GEN and 8 ng/ml of BP A achieving similar effect to that of estradiol at 1 nM. Expression of the estrogen responsive gene, pS2 was also induced in MCF-7 cells by treatment with genistein at dose as low as 1 nM and BPA at dose as low as 4 ng/ml. Using 21 day-old ovariectomized nude mice, we examined end-bud formation and mammary gland development after treatment with bisphenol A or genistein. Compared with untreated control, mammary gland development and end-bud formation were significantly increased in mice fed genistein or bisphenol A (p<0.05). Taken together, it is concluded that GEN and BP A can act as an estrogen agonist resulting in cell proliferation and induction of the estrogen responsive pS2 gene in MCF-7 cells in vitro and in athymic mice in vivo, respectively. Therefore, it is suggested that GEN and BP A might modulate human endocrine system and these compounds might be considered as a endocrine modulator at the low levels of doses.

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Preparation of Polystyrene Beads by Suspension Polymerization with Hydrophobic Silica as a Stabilizer in Aqueous Solution (소수성 실리카를 안정제로 이용하는 수용액 상에서의 현탁중합법에 의한 폴리스티렌 입자 합성)

  • Park, Moon-Soo
    • Polymer(Korea)
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    • v.30 no.6
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    • pp.498-504
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    • 2006
  • A suspension polymerization of styrene In aqueous phase was employed to study if polystyrene particles ranging from 1 to $20{\mu}m$ can be produced. Hydrophobic silica was selected as a stabilizer and azo-bisisobutyronitrile (AIBN) as an initiator. Polymerization reaction was carried out at a selected temperature in the range of $65{\sim}95^{\circ}C$. Stabilizer concentration was varied from 0.17 to 3.33 wt% compared to the water while the concentration of the initiator was raised from 0.13 to 6.0 wt% compared to the monomer. Dispersion of hydrophobic silica into the water phase was achieved by precise control of pH. Optimum dispersion of silica was obtained at pH 10. Average particle diameter decreased with increasing amounts of stabilizer concentration initially, exhibiting the minimum average diameter at 1.67 wt% of stabilizer concentration, after which it started to Increase. It is speculated that an excessive presence of stabilizer encouraged a secondary reaction in the reaction medium, which led to particle agglomeration, and as a result an increase in average particle diameter. Molecular weight was found to be independent of stabilizer concentration between 0.13 and 1.00 wt% whereas, it increased when stabilizer concentration exceeded 1.67 wt%. Variation of molecular weight was probably caused by the reduced activity and efficiency of initiator due to the high concentration of silica, and the secondary reaction in the reaction medium, as well. An increase in the Initiator concentration and/or reaction temperature resulted in an increase in both reaction rate and particle diameter. Consequently, we have confirmed that spherical polystyrene particles with $1{\sim}20{\mu}m$ in diameter can be prepared by careful selection of the concentration of stabilizer, initiator, pH and reaction temperature.

Oestrogenic Activity of Parabens In Vitro Estrogen Assays (에틸, 프로필, 이소프로필, 부틸, 이소부틸 파라벤의 In Vitro 검색시험 연구에서의 내분비독성)

  • Lee Sung-Hoon;Kim Sun-Jung;Park Jung-Ran;Jo Eun-Hye;Ahn Nam-Shik;Park Joon-Suk;Hwang Jae-Woong;Jung Ji-Youn;Lee Yong-Soon;Kang Kyung-Sun
    • Journal of Food Hygiene and Safety
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    • v.21 no.2
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    • pp.100-106
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    • 2006
  • The use of underarm and body care cosmetics with oestrogenic chemical excipients (particularly the parabens) and the hypothesized association with breast cancer incidence, particularly in women. It is noted that the type of cosmetic product is irrelevant (e.g. antiperspirant/deodorant versus body lotion, moisturizers or sprays versus creams) and attention must focus on issues of actual exposure to chemicals through continued dermal application of body care products and the endocrine/hormonal activity and toxicity of the chemicals in the formulations. To evaluate the estrogenic activities of parabens such as ethylparaben, butylparaben, propylparaben, isobutylparaben and isopropylparaben, we used recombinant yeasts containing the human estrogen receptor [Saccharomyces cerevisiae ER+LYS 8127], human breast cancer MCF-7 cell lines and human estrogen receptor ${\alpha}\;and\;{\beta}$. In E-screen assays, isopropylparaben is the most estrogenic paraben, and in ER competition assay, isobutylparaben is the most estrogenic paraben. We evaluated isopropylparaben was most active in the recombinant yeast assay, followed by propylparaben, ethylparaben, isobutylparaben and butylparaben. Results from this study demonstrate that parabens are observed in human endocrine system. Therefore, we have shown that the parabens is induced the estrogenic activities similar to $17{\beta}$-estradiol and Bisphenol-A.

Clinical Characteristics of precocious puberty girls and Comparison Analysis of GnRH Test results with Diagnosis type (성조숙증 여아들의 임상적 특징 및 진단별 성선자극호르몬 분비호르몬 GnRH (Gonado Tropin Releasing Hormone) 검사결과의 비교분석평가)

  • Kim, Jung-In;Kwon, Won-Hyun;Moon, Ki-Choon;Lee, In-Won
    • The Korean Journal of Nuclear Medicine Technology
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    • v.20 no.2
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    • pp.54-61
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    • 2016
  • Purpose Precocious Puberty is defined as the development of secondary sexual characteristics in girls younger than 8 years, and boys 9 years. Cause premature closure of the epiphysis is a disease that eventually decreases the final adult height. In this study, we retrospectively analyzed to evaluate the diagnostic difference the GnRH (Gonado-tropin-releasing Hormone) stimulation test results with medical records of precocious puberty in girls. Materials and Methods From February 2015 to December 2015 it was enrolled in the girls 118 people who visited the Seoul National University Bundang Hospital, Pediatrics, Endocrinology Internal Medicine. True precocious puberty group (n=57), early puberty group (n=39), were divided into Premature thelarche (n=22) group. A Tanner stage, chronological age, bone age, height, body weight for each group was determined by examining the mean${\pm}$standard deviation. GnRH test result was compared LH (Basal, 30 min, 45 min, 60 min), FSH (Basal, 30 min, 60 min) for each group, Each group LH, FSH Peak value distribution, the mean${\pm}$standard deviation was calculated for the peak LH/LH basal ratio, peak LH/Peak FSH ratio. The significance probability (P-value) between the value of each third group was determined. Results The average height of the true precocious puberty group $131{\pm}14.85$, the mean weight was $28.80{\pm}4.93$, the average chronological age $7.1{\pm}0.81$, the mean bone age was $9.9{\pm}0.9$, The average height of early puberty group was $134{\pm}5.10$, the average weight $28.50{\pm}4.43$, the average chronological age $8.05{\pm}0.03$, the mean bone age was $10.0{\pm}0.62$, The average height of Premature thelarche $129{\pm}6,01$, the average weight was $28.65{\pm}5.98$, the average chronological age $7.02{\pm}0.58$, the mean bone age was $8.04{\pm}1.29$. There was no significant difference when compared to the height and weight. There was a significant difference between the groups in the chronologic age and bone age difference (P <0.0002) True precocious puberty group showed peak LH levels at 30'(82.5%), 45'(12.3%), 60'(5.3%), in Peak FSH 30'(8.8%), 60'(91.2%). Early Puberty group showed high values in Peak LH at 30'(79.5%), 45'(17.9%), 60'(2.6%), in peak FSH levels at 30'(7.7%), 60'(92.32%). In Premature thelarche Group it showed the Peak LH levels at 30'(30%), 45'(59%), 60'(9.09%), Peak FSH levels at 30'(0%) 60'(100%). When compared with the The Peak LH/basal LH ratio, True precocious puberty group was $19.09{\pm}17.15$, early puberty group was $15.23{\pm}10.88$, Premature thelarche group showed significant differences between the three groups as $4.93{\pm}4.36$.(P <0.0001) LH Peak/FSH Peak ratio, true precocious puberty group was $1.222{\pm}0.77$, early puberty group was $1.34{\pm}1.23$, Premature thelarche group showed significant differences between the three groups as $0.3{\pm}0.09$(P <0.0001) Conclusion In order to diagnose the true precocious puberty have a diagnostic value when the LH peak after GnRH stimulation is increased by more than two to three times compared to baseline or a predetermined level or more than 5~10 IU/L increases. GnRH Test is a test for a long time and the patient discomfort due to repeated blood sampling, but the hypothalamus-pituitary gland- gonad axis activity evaluate and is the most basic accurate test in the differential diagnosis of precocious puberty disorders.

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