• Title/Summary/Keyword: Hormonal

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Transformation of Potato using the Phosphinothricin Acetyltransferase Gene as the Selectable Marker Gene (감자의 형질전환을 위한 표지유전자로서 Phosphinothricin Acetyltransferase 유전자의 이용)

  • Jeong, J.H.;Yang, D.C.;Bang, K.S.;Han, S.S.
    • Korean Journal of Weed Science
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    • v.18 no.3
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    • pp.205-213
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    • 1998
  • This experiment was carried out to produce herbicide resistant potatoes hawing only chimeric phosphinothricin acetyltransferase (PAT) genes without using antibiotic selectable marker. The pDY502 vector having only PAT gene was reconstructed for transformation of potato. The reconstructed vector was introduced to Agrobacterium tumefaciens MP90 disarmed, and they were used for potato transformation. Hormonal requirement for plant regeneration from leaves and stem explants of potato was investigated. From this experiment, MS medium treated with IBA 0.1 mg/L + BA 0.5 mg/L was the best for potato regeneration, and the ratio of shoot regeneration was 54% for leaf and 46% for stem in that condition. For transformation, explants of potato leaves and stems were cocultured with A. tumefaciens MP90 containing reconstructed vector harvoring only PAT gene. When the potato explants were placed on various concentrations of bialaphos and all the potato explants were dead on medium with over 5.0mg/L bialaphos. By this selection methods, the explants cocultured with Agrobacterium produced the putative transgenic shoots on medium with 5mg/L bialaphos treatment after 3-4 weeks. Second selection was performed by transferring the shoot tips of putative transgenic to medium containing 20mg/L of bialaphos. The shoot tips grew well on the second selection medium, indicating the production of successful transgenic plants. But normal shoots were dead in same cytotoxic medium. Incorporation of the PAT gene into transgenic potatos were confirmed by PCR analysis of DNA and Southern hybridization. These results show that the PAT gene can serve as a selectable marker and herbicide resistant genes for transformation of potato.

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Can high serum anti-M${\ddot{u}}$llerian hormone levels predict the phenotypes of polycystic ovary syndrome (PCOS) and metabolic disturbances in PCOS patients?

  • Hwang, Yu Im;Sung, Na Young;Koo, Hwa Seon;Cha, Sun Hwa;Park, Chan Woo;Kim, Jin Yeong;Yang, Kwang Moon;Song, In Ok;Koong, Mi Kyoung;Kang, Inn Soo;Kim, Hye Ok
    • Clinical and Experimental Reproductive Medicine
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    • v.40 no.3
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    • pp.135-140
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    • 2013
  • Objective: To evaluate correlations between serum anti-M${\ddot{u}}$llerian hormone (AMH) levels, phenotypes of polycystic ovary syndrome (PCOS), obesity, and metabolic parameters in patients with PCOS. Methods: A total of 175 patients with PCOS were diagnosed according to the Rotterdam Consensus were included. Exclusion criteria were age over 40, FSH>25 mIU/mL, and 17a-OHP>1.5 ng/mL. The Phenotypes of PCOS were divided into a severe form (oligo-anovulation, ANOV/hyperandrogenism/polycystic ovary morphology [PCOM]; n=59) and a mild form without HA (ANOV/PCOM, n=105). The serum AMH levels were classified into 3 groups (<5 vs. 5-10 vs. >10 ng/mL). Obesity was defined as body mass index (BMI) ${\geq}25kg/m^2$ (n=34). Results: The mean age was $25.9{\pm}5.7$ year and mean AMH level was $10.1{\pm}5.4$ ng/mL. The BMI ($kg/m^2$) was higher in group 1 ($24.2{\pm}6.3$) than in group 2 ($21.9{\pm}4.3$, p=0.046) or group 3 ($21.6{\pm}3.3$, p=0.019). There was no difference among the three groups in age, menstrual interval, antral follicle counts, androgens, or other metabolic parameters. The obesity group showed significantly lower AMH ($7.7{\pm}3.9$ ng/mL vs. $10.7{\pm}5.6$ ng/mL), p=0.004) and low-density lipoprotein levels ($93.1{\pm}21.2$ mg/dL vs. $107.5{\pm}39.3$ mg/dL, p=0.031), and showed higher total T ($0.74{\pm}0.59$ L vs. $0.47{\pm}0.36$ ng/mL, p=0.001), free T ($2.01{\pm}1.9$ vs. $1.04{\pm}0.8$ pg/mL, p=0.0001), and free androgen index ($6.2{\pm}7.9$ vs. $3.5{\pm}3.0$, p=0.003). After controlling for age factors and BMI, the serum AMH levles did not show any significant correlations with other hormonal or metabolic parmeters. Conclusion: For PCOS patients under the age 40, serum AMH is not negatively correlated with age. High serum AMH levels can not predict the phenotype of PCOS and metabolic disturbances in PCOS patients in the non-obese group. Further study might be needed to define the relation more clearly.

Studies on the postnatal development of the Leydig cell in rabbits (토끼에서 출생 후 고환간질세포의 발생에 관한 연구)

  • Tae, Hyun-Jin;Park, Young-Jae;Kang, Hyung-Sub;Kim, Nam-Soo;Park, Sang-Youel;Yang, Hong-Hyun;Ahn, Dong-Choon;Kim, In-Shik
    • Korean Journal of Veterinary Research
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    • v.45 no.3
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    • pp.325-334
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    • 2005
  • Changes in the rabbit Leydig cell from birth to adulthood were studied in New Zealand white rabbits of 1, 7, 21, 35, 49, 70, 105, 147, 196, and 252 days (n = 8 rabbits per group) of age. The objectives of this study were to understand the fate of the fetal Leydig cells, to determine the changes in serum testosterone levels, and leutenizing hormone-stimulated testosterone production per testis in vitro, and to quantify adult Leydig cells by number and average volume with age. Testes of rabbits were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative (stereological) morphological studies were performed. Testosterone levels in the incubation medium of luteinizing hormone-stimulated (100 ng/ml) testosterone secretion per testis in vitro, and in serum were determined by radioimmunoassay. The average volume of a testis of 1-day-old rabbits was determined as $0.0073cm^3$ and the parameter increased linearly from birth to 252 days ($3.93cm^3$). The volume density of the seminiferous tubules increased with age from 33.76% at day 1 to 88.2% at day 252. The volume density of the interstitium represents 66.24% of the testicular parenchyma at day 1. This proportion progressively diminished during development to reach a value of 11.8% at day 252. The volume density of Leydig cells increased almost linearly from birth (0.001%) to 252 days (2.62%). Leydig cell mass per testis increases from 0.0012 mg to 0.25 mg between days 1 and 35, from 2.66 mg to 44.3 mg between days 49 and 105 and from 65.42 mg and 102.9 mg between days 147 and 252. The absolute numbers of adult Leydig cells per testis increased linearly from birth to 252 days. The average volume of adult Leydig cell on days 1, 7, 21 and 35 was not significantly different; a gradual and continued increase was observed thereafter, reaching a 3-fold increase at 196 and 252 days. Serum testosterone concentrations were not significantly different at day 1 compared days 7, 21, 35. Significant increases were observed at days 49 and 70. Values at days 70 and 105 and days 147, 196, and 252 were not significantly different. LH-stimulated testosterone production per testis in vitro was significantly different at day 1 compared days 7, 21, 35. Significant increases were observed at days 49 and 70. Hormonal values at days 105, 147, 196, and 252 were not significantly different. These data suggested Leydig cell developmental phase can be classified: a neonatal phase (1-7 days), a prepubertal phase (14-49 days) and an adult phase (70-252 days). Immature and mature adult Leydig cells, initially detected at days 7 and 49, respectively, and mature adult Leydig cells were abundant Leydig cell type according to the number and absolute volume per testis form day 49 onwards.

Study of Rat Mammary Epithelial Stem Cells In Vivo and In Vitro (생체 및 시험관에서 유선 상피 모세포의 분리와 동정)

  • Nam Deuk Kim;Kee-Joo Paik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.3
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    • pp.470-486
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    • 1995
  • Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.

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Callus Initiation and Organ Formation from in vitro Culture of Pelargonium spp. (In Vitro Culture에 의한 Pelargonium spp.의 Callus 유도 및 기관분화에 관하여)

  • Lee, Chun-Ha;Chung, Hae-Joon
    • The Journal of Natural Sciences
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    • v.4
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    • pp.143-159
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    • 1991
  • In order to investigate the micropropagation of Pelargonium, 2 cultivars of P. peltatum 'Pouletta' and P. zonale 'Pinto Red' were cultured in vitro on the MS basal medium supplemented with various concentrations of growth regulators. It attempted to study the induction of callus and the differentiation of organs from leaf disc, petiole segments, stem segments. hypocotyle segments and flower stalk segments. The results are summarized as follows; A. As for the initiation of callus, stem explant was proved to be the most suitable one among various explants of P. zonale 'Pinto Red'. The medium was supplemented with 1.0mg/1 BAP and 1.0mg/1 NAA. As NAA concentration increased, callus formation was enhanced, but higher concentration of NAA inhibited callus fromation. Leaf and hypocotyle explants showed less callus formation than stem and petiole explants. B. In P. zonale 'Pinto Red' petiole culture, the condition of cullus culture such as hormone concentration resulted in affecting shoots differentiation. The best result of shoots formation from the callus reculture were obtained from the combination of 0.5-1.0mg/1 BAP and 0.1-1.0mg/1 NAA when the callus was cultured in 1.0mg/1 BAP and 0.05mg/1 NAA. When the callus was cultured in medium without BAP, the shoot was not differentiated in subculture regardless to BAP and NAA concentration. and only callus was formed. C. Poly-phenol substance was observed in MS medium supplemented without PVP, in which callus was not formed from the leaf of P. peltatum 'Rouletta'. Polyphenol substance was not observed in MS medium supplemented with PVP, in which callus formation was increased. D. The callus formation of P. peltatum 'Rouletta' showed the stem explant being best result. The best result particularly in the stem explant among others. The optimal hormonal concentration was 0.1mg/1 NAA and 5.0mg/1 BAP. The shoot formation was observed at 0.05mg/1 NAA and 1.0mg/1 BAP, 0.1mg/1 NAA and 5.0mg/1 BAP. The shoot was malformed and the tissue recultured turned necrotic.

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Anti-Stress Effect of Punica granatum L. Extract against Sleep Deprivation-Induced Impairment (석류 열수 추출물의 수면박탈을 유도한 Rat 모델에서의 항스트레스 효과)

  • Na, Ju-Ryun;Kim, Sunoh;Jo, Ara;Bae, Donghyuck;Oh, Kyo-Nyeo;Kim, Yong Jae;Lee, Yoo-Hyun;Jun, Woojin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.11
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    • pp.1533-1543
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    • 2016
  • The anti-stress effects of Punica granatum L. (family Lythraceae, PG) on $H_2O_2$/corticosterone (CORT)-induced stress in cells and sleep-deprived rats were investigated. The PG extract showed neuroprotective effects in SH-SY5Y cells against $H_2O_2$/CORT-induced stress. Sleep deprivation led to behavioral, hormonal, and biochemical alterations in the animal model. The effects of P. granatum on physiological, behavioral, and biochemical parameters aggravated by sleep deprivation were investigated. Sleep deprivation impaired physiological (survival, body weight, and drowsiness scores) and behavioral (rotarod, passive avoidance, hot hyperalgesia, and Y maze) parameters as well as biochemical factors (cortisol, serotonin, dopamine, testosterone, and growth factor I contents in serum). These parameters were significantly recovered by PG extract in a concentration-dependent manner. The PG extract also enhanced catalase, superoxide dismutase, and non-enzymatic antioxidative activities such as glutathione compared to sleep-deprived rats. On the basis of these results, our findings suggest that Punica granatum prevents impairment of body functions induced by sleep deprivation and related oxidative damage.

Monocentric Derivative Y Chromosome with Duplication of the SRY Gene in an Azoospermic Male (무정자증 불임남성에서 관찰된 SRY 유전자의 중복을 동반한 일동원체성 derivative Y 염색체)

  • Choi, Eun-Young;Lee, Bom-Yi;Park, Ju-Yeon;Lee, Yeon-Woo;Oh, Ah-Rum;Lee, Shin-Young;Kim, Shin-Young;Han, You-Jung;Lee, Mee-Bum;Ryu, Hyun-Mee;Seo, Ju-Tae;Park, So-Yeon
    • Journal of Genetic Medicine
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    • v.7 no.2
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    • pp.160-164
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    • 2010
  • Structural abnormalities of the Y chromosome affect normal testicular differentiation and spermatogenesis. The present case showed a rare monocentric derivative Y chromosome with partial duplication of Yp including the SRY gene and deletion of Yq12 heterochromatin. The karyotype was 46,X,der(Y)(pter${\rightarrow}$q11.23::p11.2${\rightarrow}$pter).ish der(Y)(DYZ3+,DYZ1-,SRY++), confirmed through a FISH study. Even though the patient possessed an abnormal Y chromosome, testicular biopsy showed normal testicular volumes in the proband, with gonadal hormonal levels in the normal range but bilateral varicocele and hypospermatogenesis. We speculate that the abnormal Y chromosome arose from sister chromatids during Y chromosome recombination or intra chromosomal NAHR (non-allelic homologous recombination) during meiosis in the patient's father or in the very early stages of embryogenesis. The derivative Y chromosome might interfere in the meiotic stage of spermatogenesis, leading to the developmental arrest of germ cells. The present case illustrates that the infertility phenotype can have various causes. Also, it emphasizes the importance of accurate and various genetic analyses and could aid in male infertility treatment.

Changes in Endogenous Abscisic Acid, Jasmonic Acid and Sucrose Content during Bulb Development in the Cold-type Cultivar of Garlic (Allium sativum L.) of Korea (한지형 마늘의 인경 발육과정에서 식물내생호르몬 Abscisic Acid, Jasmonic Acid 및 당 함량변화)

  • Sohn, Eun-Young;Kim, Yoon-Ha;Kim, Jung-Tae;Jang, Soo-Won;Lee, In-Jung
    • Horticultural Science & Technology
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    • v.29 no.1
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    • pp.1-9
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    • 2011
  • This study was performed to investigate the role of plant growth substances on the bulbing of cold type of garlic (Allium sativum L. cv. Uiseongmaneul) during long and short day conditions. The change in endogenous plant hormones such as abscisic acid (ABA), jasmonic acid (JA) and sugar contents in leaf blade and sheath was examined during the growth stage from bulb differentiation (starting at April 16) to bulbing (April 24 to May 18) in cold type of garlic. In the long day condition, ABA contents were higher than short day condition and ABA contents of leaf sheath were higher than leaf blade. ABA contents of growth stage in garlic were not changed during growth stage from April 16 to May 2, however it rapidly increased during bulbing (May 2 to May 18). On the other hand, endogenous JA contents in short day condition did not change in long day condition, it increased from April 16 to May 2. JA contents in the leaf sheath (33.85-62.04 $ng{\cdot}g^{-1}$ DW) were higher than leaf blade (15.39-30.04 $ng{\cdot}g^{-1}$ DW). These results showed that garlic bulb differentiation and bulbing was induced by JA in leaf sheath. In long day condition, total sugar contents in the leaf blade were increased from bulb differentiation (April 16) to bulbing (May 4) and it was decreased during bulbing (May 4 to May 18) while the total sugar content in leaf sheath were gradually increased from bulb differentiation (April 16) to bulbing (May 18). In conclusion, our results showed that there is a significant correlation between the bulb development of garlic and hormonal content in the leaf sheath.

Skin Coloration and Endogenous Hormonal Changes of 'Kyoho' Grape by High Temperature at Veraison (변색기 고온에 의한 포도 '거봉'의 과피 착색 및 내생 호르몬 변화)

  • Ryu, Suhyun;Cho, Jung-Gun;Jeong, Jae Hoon;Lee, Seul-Ki;Han, Jeom Hwa;Kim, Myung-Su
    • Journal of Bio-Environment Control
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    • v.28 no.3
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    • pp.234-242
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    • 2019
  • We analyzed the anthocyanin accumulation, abscisic acid (ABA), gibberellic acid (GA) contents and metabolic genes expression in berry skins under high temperature (High T) at veraison, in order to investigate the cause of bad coloration of 'Kyoho' grape due to High T in summer season. The coloration of 'Kyoho' grapes was stopped by High T for 10 days from veraison, and the fruit quality was not affected except skin color. Total anthocyanin of skins was decreased by High T treatment and malvidin and peonidin were decreased compared to control. In berry skins, ABA content did not decrease by High T treatment, but it was rather higher than that of control. GA content was increased about two times compared to the control after 10 days of High T treatment, which caused decreased ratio of ABA/GA. Analysis of expression of anthocyanin biosynthetic genes showed that the early biosynthetic genes were not affected by High T and the expression of UFGT was decreased by temperature treatment. ABA biosynthetic gene expressions were not affected by High T and the expression of GA20ox1 and GA2ox1/2, which are known to regulate the biosynthesis and inactivation of GA, were increased and decreased by High T, respectively. Therefore, the bad coloration of 'Kyoho' grapes under the High T at veraison was due to inhibition of anthocyanin biosynthesis of skin, and it was suggested that the anthocyanin biosynthesis was controlled by the ratio of ABA and GA rather than ABA content.

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.