• Title/Summary/Keyword: Group maturation

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Changes in gene expression associated with oocyte meiosis after $Obox4$ RNAi

  • Lee, Hyun-Seo;Kim, Eun-Young;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.2
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    • pp.68-74
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    • 2011
  • Objective: Previously, we found that oocyte specific homeobox (Obox) 4 plays significant role in completion of meiosis specifically at meiosis I-meiosis II (MI-MII) transition. The purpose of this study was to determine the mechanism of action of $Obox4$ in oocyte maturation by evaluating downstream signal networking. Methods: The $Obox4$ dsRNA was prepared by $in$ $vitro$ transcription and microinjected into the cytoplasm of germinal vesicle oocytes followed by $in$ $vitro$ maturation in the presence or absence of 0.2 mM 3-isobutyl-1-metyl-xanthine. Total RNA was extracted from 200 oocytes of each group using a PicoPure RNA isolation kit then amplified two-rounds. The probe hybridization and data analysis were used by Affymetrix Gene-Chip$^{(R)}$ Mouse Genome 430 2.0 array and GenPlex 3.0 (ISTECH, Korea) software, respectively. Results: Total 424 genes were up (n=80) and down (n=344) regulated after $Obox4$ RNA interference (RNAi). Genes mainly related to metabolic pathways and mitogen-activated protein kinase (MAPK) signaling pathway was changed. Among the protein kinase C (PKC) isoforms, PKC-alpha, beta, gamma were down-regulated and especially the MAPK signaling pathway PKC-gamma was dramatically decreased by $Obox4$ RNAi. In the cell cycle pathway, we evaluated the expression of genes involved in regulation of chromosome separation, and found that these genes were down-regulated. It may cause the aberrant chromosome segregation during MI-MII transition. Conclusion: From the results of this study, it is concluded that $Obox4$ is important upstream regulator of the PKC and anaphase-promoting complex action for maintaining intact germinal vesicle.

Characterization of ovarian culture in vitro and sex steroids in vivo by recombinant eel gonadotropin treatments in the eel Anguilla japonica

  • Kim, Dae-Geun;Kim, Jung-Hyun;Baek, Hea-Ja;Kim, Shin-Kwon;Min, Kwan-Sik;Kim, Dae-Jung
    • Fisheries and Aquatic Sciences
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    • v.25 no.1
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    • pp.12-19
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    • 2022
  • In the present study, we investigated the effects of recombinant eel gonadotropins (rec-GTHs) on maturation induction in immature ovarian culture in vitro and sex steroid hormones in vivo in the Japanese eel Anguilla japonica. To study the in vitro effects of rec-GTHs on estradiol-17β (E2) production in immature ovarian tissues, ovarian tissues were incubated with different doses of rec-follicle-stimulating hormone (rec-FSH) or rec-luteinizing hormone (rec-LH). The results revealed that the E2 levels in the rec-FSH (0.1, 0.5, or 1 ㎍/mL)- and rec-LH (0.1 or 0.5 ㎍/mL)-treated groups were significantly higher than those in the female eels from the control group. Furthermore, to investigate the in vivo effects of rec-GTHs on the gonadosomatic index (GSI) and plasma sex steroid hormone levels, the eels were injected intraperitoneally with eel's ringer (control), salmon pituitary extract (SPE; for female eels), human chorionic gonadotropin (hCG; for male eels), rec-FSH, rec-LH, and rec-FSH + rec-LH once a week. The results revealed that except for the SPE and the hCG groups, none of the groups exhibited a significant difference in GSI values. However, in vivo plasma E2 levels increased at the end of 4 weeks after rec-FSH treatment in female eels. Based on these results, it is suggested that rec-GTHs may have a positive effect on sexual maturation in female eels; however, further studies on complementary rec-protein production systems and additional glycosylation of rec-hormones are needed to elucidate hormone bioactivity in vivo and in vitro.

Effects of Massage and Attachment Promotion Program on Social Maturity, Child Autism and Attachment of Children with Autism and Their Mothers

  • Lee, Hae-Kyung
    • Child Health Nursing Research
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    • v.14 no.1
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    • pp.14-21
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    • 2008
  • Purpose: The aims of this study were to evaluate effects of massage therapy and the attachment promotion program with autistic children. Method: A quasi-experimental design was conducted with a convenience sample of 44 autistic children. The experimental group (n= 23) attended the massage therapy and the attachment promotion program for 4 months. The control group (n=21) only had the attachment promotion program. Measurements were made twice for both groups using the social maturity scale and child autism rating scale. Recording of mother-child attachment was done for 15 minutes in only the experimental group twice using video equipment. Results: After 4 months of interventions, there was significant difference on social maturity (F=9.01, p=.005) between the groups. However, there was no significant difference on CARS ((F=2.47, p=.124). The total scores of mother-child attachment between pre- and post-interventions showed a significant difference (Z=-3.42, p=.001). Conclusion: The results showed that massage therapy and the attachment promotion program might be an effective way for providing a chance to increase social maturation and to increase attachment between mother and autistic child.

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Effect of Prepubertal Exposure to Di(2-ethylhexyl)phthalate on the Maturation of Rat Seminal Vesicles and Prostate Glands (사춘기 전 수컷 흰쥐의 저정낭과 전립선의 성숙에 미치는 Di(2-ethylhexyl) phthalate(DEHP)의 영향)

  • Heo, Hyun-Jin;Lee, Won-Yong;Yoon, Yong-Dal;Choi, Donchan;Lee, Sung-Ho
    • Development and Reproduction
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    • v.12 no.3
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    • pp.251-259
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    • 2008
  • The plasticizer di(2-ethylhexyl)phthalate(DEHP) is one of the most well known endocrine disrupting chemicals (EDCs) because of its strong anti-androgenic effects on the reproductive and developmental process in male rodents and human. The present study was performed to examine whether prepubertal exposure to DEHP can make any alteration during the maturation of accessory sex organs in male rats. As a result, there was no significant change in body weights, serum T levels and tissue weights except of seminal vesicle and ventral prostate in DEHP-treated animals compared to vehicle-treated ones. The seminal vesicle weights in high-dose group (200 mg/kg) were significantly lower than those from the control group (p<0.05), and ventral prostate weights were significantly lower than those from the control group (p<0.05) in both low-dose (20 mg/kg) and high-dose group. Histological studies revealed that the seminal vesicles from DEHP-treated groups showed reduced areas of mucosal folds. Pseudostratified columnar epithelia were observed in the ventral prostates of DEHP-treated samples while cuboidal epithelia were found in the control group. The transcriptional activities of ER-$\alpha$ in seminal vesicle from high-dose group (p<0.05) were significantly higher than those from the control group, and ER-$\beta$ expression was significantly decreased in low-dose group (p<0.05) compared to the control. In ventral prostate, ER-$\beta$ mRNA levels from low-dose group (p<0.05) were significantly lower than those from the control group, and significantly increased in high-dose group (p<0.01). AR expressions, however, were not significantly different in all experimental groups of both seminal vesicle and ventral prostate. In conclusion, the present study demonstrated that (i) adverse effect (s) of DEHP on sexual maturation during prepubertal period could be limited, (ii) seminal vesicle and prostate gland were sensitive targets to DEHP in prepubertal rats and (iii) the deleterious effects of DEHP might be mediated through ER-associated mechanism.

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Porcine Oocytes with Meiotic Competence are Synchronized Early Stage of Germinal Vesicle (GV)

  • Park, Kwang-Wook
    • Reproductive and Developmental Biology
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    • v.34 no.3
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    • pp.169-173
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    • 2010
  • Correlations between cumulus cells and germinal vesicle (GV) chromatin configuration were examined in porcine oocytes. Cumulus-oocyte complexes (COCs) were collected from 2~6 mm follicles and divided into three categories according to cumulus cell morphology. "A" group was compacted COCs with more than three cumulus cell layers. "B" group was COCs with less cumulus cell layers than "A" group. "C" group was COCs with one or less layer of cumulus cells. Cumulus cells were removed 0.1% hyaluronidase, and denuded oocytes were stained with Hoechst 33342. GV chromatin configuration was classified into GV-Con and GV-Dis. GV-Con meant that a nucleus was surrounded by condensed chromatin in a ring. GV-Dis meant that filamentous chromatin clumps were distributed in nucleus. The proportion (80.2%) of GV-Con in "A" group was significantly higher than "B" (62.0%) or "C" (44.9%). The proportion (55.1%) of GV-Dis in "C" group was significantly higher than "A" (19.8%) or "B" (38.0%). The meiotic competence of COCs was examined after 44 h culture. The proportion (90.0%) of oocytes reaching to metaphase II (M-II) in "A" group was significantly higher than "B" (76.5%) or "C" (45.5%). In conclusion, oocytes with good quality cumulus cell layers are synchronized early GV stage, and early GV stage is important for meiotic competence in pigs.

Relations between Testicular Feminization and Seminiferous Tubule (Testicular Feminization과 세정관과의 관계)

  • Lee, Sok-Woo;Lee, Kwang-Ho
    • The Korean Journal of Zoology
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    • v.29 no.1
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    • pp.61-69
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    • 1986
  • Teter and Boczkowski (1966) classified testicular feminization syndrome into two groups; One with no pubic hair and normal clitoris (A group) and the other with pubic hair and enlarged clitoris (B group). However, the patient discussed in this paper shows a mid-form of these two groups (C group). To find out differences among A, B and C groups, we have observed the micro-structure of testes removed from her inguinal canal and analyzed the chromosomes obtained from testicular and blood culture. Her seminiferous tubules show some differences in the maturation and distribution of Leydig and Sertoli cells and in the thickening of basement membrane as compared with group A and B. We suppose that such differences are responsible for the morphological changes of clitoris and pubic hair.

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A Study on the 6-Week Cleansing Effect of Placenta Cold Process Soap (자하거저온숙성비누의 6주간 세안효과에 대한 연구)

  • Jin Suk, Koo
    • The Korea Journal of Herbology
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    • v.38 no.2
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    • pp.9-16
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    • 2023
  • Objectives : Many people use natural soap instead of commercially available bar type general soap for skin care. They tend to prefer cp (cold process) type soaps that go through a sufficient maturation process in formulation. The purpose of this study was to investigate the efficacy of PCPS (placenta cp soap). Methods : The researcher selected 36 subjects, 28 as the experimental group and 8 as the control group. I asked participants to wash their face twice a day in the morning and evening, and the period was set for 6 weeks. The researcher had the people who were selected as the control group use a commercially available foam cleanser, meanwhile the experimental group use PCPS. They performed a skin test before the start of the test, and the change status was continuously investigated after 2 weeks, 4 weeks, and 6 weeks after using the soap. Results : In the case of U zone oil and pores, there was a significant decrease and in the case of skin tone, there was a significant increase in the group of using PCPS. In T zone oil change, there was no significant effect in both the experimental group and the control group. Conclusion : In the case of PCPS, it is recommended to use it in the effect of reducing the size of pores and increasing skin tone, but it can reduce u-zone oil content.

Effects of FGF on Embryonic Development In Vitro in Hanwoo COCs (한우 난구 복합체의 체외발생에 있어서 FGF(Fibroblast Growth Factor)가 미치는 영향)

  • Choi S.H.;Cho S.R.;Kim H.J.;Choe C.Y.;Han M.H.;Son D.S.;Chung Y.G.;H. Hoshi
    • Journal of Embryo Transfer
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    • v.21 no.2
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    • pp.157-162
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    • 2006
  • It is well known that unidentified factors in sera, hormones and growth factors promote the proliferation of granulosa cells and nuclear maturation of bovine COCs (cumulus oocytes complexes) in vitro. Attempts had been developed the simple composition of culture media and similar system to in vivo conditions has been applied. In the present study, we investigated the effect of FGF (fibroblast growth factor) on in vitro maturation and in vitro development of Hanwoo COCs. When the COCs were matured in HPM 199 (Inst. of Functional peptide, Japan) containing 0.1, 1 and 10 ng/ml FGF for 24 hr, maturation rates to metaphase II ($70.0{\sim}75.0%$) were significantly higher (p<0.05) than that of control group (0 ng/ml FGF, 37.5%). When matured COCs with FGF were cultured in maturation medium after in vitro fertilization, developmental rates to blastocysts were 9.5, 0 and 2.9%, respectively, compared to 25.0% of the control group (p<0.05). When the matured COCs with FGF were cultured in HPM 199 (IFP971, Inst. of Functional peptide, Japan) containing 10% FBS, 0.8% BSA or 0.1% PVA (polyvinyl alcohol), the blastocyst formation rates were 12.4, 12.8 and 8.5%, respectively, while the rates of matured COCs with FGF and cultured with IVMD and IVD (Inst. of Functional peptide, Japan) without serum were 38.4% and 34.8%, respectively (p<0.05). These results suggested that FGF is available for in vitro maturation of bovine COCs and is not suitable for in vitro development, but further investigation would be need for finding the synergistic autocrine/paracrine fashion of other growth factors in early bovine embryo development.

Determination of the Granulosa Cell-Specific Endothelin Receptor A Deletion on Ovarian Function

  • Cho, Jong-Ki
    • Journal of Embryo Transfer
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    • v.29 no.2
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    • pp.195-200
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    • 2014
  • Endothelin 2 (EDN2) induces follicular rupture by constricting periovulatory follicles. In this study, it was investigated the mechanisms of EDN2 action on follicular rupture with respect of receptor using the conditionally granulosa cell specific EDN2 receptor type A (ETa) KO mice (gcETaKO; $ETa^{flox/-}{\cdot}Amhr2^{Cre}$). It was generated the gcETaKO mice by breeding with $ETa^{flox/-}$ mice after mono-alleic ETa knockout by $ZP3^{Cre}$ and $Amhr2^{Cre}$ mice. Fertility, ovulation and maturation rates of ovulated oocytes after super ovulation were investigated in the gcETaKO mice compared with wild-type mice ($ETa^{flox/flox}$ and $ETa^{flox/-}$) as a control group. In the gcETaKO mice, normal fertility after breeding with male mice was shown compared with wild-type mice. And, there was no significant differences in ovulation rates after super ovulation, however its maturation rates was lower than that of wild type mice. These findings show that EDN2 in follicular rupture for ovulation is related with an other ETa not in granulosa cells. Further studies are needed to investigate how EDN2 is acted in ovarian follicular rupture for ovulation.

The Use of Fluorescein Diacetate (FDA) to Assess Oocyte Viability in the Bovine (FDA를 이용한 소 난포란의 생사 판정)

  • 김중계;박세필
    • Korean Journal of Animal Reproduction
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    • v.21 no.3
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    • pp.211-217
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    • 1997
  • This experiment was carried out to determine the possibility of using FDA on the selection of viable bovine immature oocytes. The results obtained in these experiments were summarized as follows; 1. The rates of nuclear maturation (metaphase II) of bovine oocytes according to FDA concentration were 92.5% (37/40) in the control and those rates in the dilution on FDA to 1:400,000, 1:800,000 and 1:1,600,000 were 74.4% (67/90), 80.3% (38/46) and 71.1% (32/45), respectively. 2. The fertilization rate ( 2-cell) in the control was 72.2% (39/54) and those rates in the dilution of FDA to 1:400,000, 1:800,000 and 1:1,600,000 were 28.8% (23/80), 66.7% (32/48) and 62.2% (28/45), respectively. In these results, a, pp.opriate concentration of FDA to bovine immature oocytes was 1:800,000. 3. Effect of FDA treatment to the blastocyst development of bovine oocytes was indicated that control was 22.2% (18/81) and FDA treatment groups which were classified to strong, partial and weak were 21.4% (36/168), 14.5% (9/62) and 0% (0/15), respectively. This result suggested that in vitro development to the blastocyst was severely reduced except strong group according to the FDA fluorescent level (P<0.05) and that using FDA is possible to select of bovine oocytes.

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