• Title/Summary/Keyword: Developmental rate

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Implications for the Predictivity of Cell-Based Developmental Toxicity Assays Developed Two Decades Apart

  • Kawamura, Satoshi;Horie, Nobuyuki;Okahashi, Noriko;Higuchi, Hashihiro
    • Toxicological Research
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    • v.35 no.4
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    • pp.343-351
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    • 2019
  • Many in vitro developmental toxicity assays have been proposed over several decades. Since the late 1980s, we have made intermittent attempts to introduce in vitro assays as screening tests for developmental toxicity of inhouse candidate products. Two cell-based assays which were developed two decades apart were intensively studied. One was an assay of inhibitory effects on mouse ascites tumor cell attachment to a concanavalin A-coated plastic sheet surface (MOT assay), which we studied in the early days of assay development. The other was an assay of inhibitory effects on the differentiation of mouse embryonic stem cell to beating heart cells (EST assay), which we assessed more recently. We evaluated the suitability of the assays for screening in-house candidates. The concordance rates with in vivo developmental toxicity were at the 60% level. The EST assay classified chemicals that inhibited cell proliferation as embryo-toxic. Both assays had a significant false positive rate. The assays were generally considered unsuitable for screening the developmental toxicity of our candidate compounds. Recent test systems adopt advanced technologies. Despite such evolution of materials and methods, the concordance rates of the EST and MOT systems were similar. This may suggest that the fundamental predictivity of in vitro developmental toxicity assays has remained basically unchanged for decades. To improve their predictivity, in vitro developmental toxicity assays should be strictly based on elucidated pathogenetic mechanisms of developmental toxicity.

Reprogramming of Cloned Embryos During Early Embryogenesis (초기 발생에 있어서 복제수정란의 리프로그래밍)

  • Han, Yong-Mahn;Kang, Yong-Kook;Koo, Deog-Bon;Lee, Kyung-Kwang
    • 대한생식의학회:학술대회논문집
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    • 2002.05a
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    • pp.11-17
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    • 2002
  • Animal clones derived from somatic cells have been successfully produced in a variety of mammalian species such as sheep, cattle, mice, goats, pigs, cat and rabbits. However, there are still many unsolved problems in the present cloning technology. Somatic cell nuclear transfer has shown several developmental aberrancies including high rate of abortion in early gestation and increased perinatal death. These developmental failures of cloned embryos may arise from abnormal reprogramming of donor genome and/or incomplete cloning procedure. We have found that overall genomic methylation status of cloned bovine embryos is quite different from that of normal embryos in various genomic regions, suggesting that the developmental failures of cloned embryos may be due to incomplete reprogramming of donor genomic DNA. Many of the advances in understanding the molecular events for reprogramming of donor genome will more clarify the developmental defects of cloned embryos.

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Nonmonotonic Effects of Chronic Low-Dose Di(2-ethylhexyl) Phthalate on Gonadal Weight and Reproduction

  • Cha, Sunyeong;Jung, Kayeon;Lee, Min Young;Hwang, Yeon Jeong;Yang, Eunhyeok;Lee, Sung-Ho;Jung, Hyo-il;Cheon, Yong-Pil
    • Development and Reproduction
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    • v.22 no.1
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    • pp.85-94
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    • 2018
  • Endocrine disruptors have been concerned in toxicology but now challenged as physiological point especially concerned with exposing dose and period. In this study the low-dose chronic administration of di(2-ethylhexyl) phthaltae (DEHP) during reproductive period was examined to evaluate the possible roles. Adult male and female CD-1 mice were exposed to DEHP with drinking water containing $133{\mu}g/L$ and $1,330{\mu}g/L$ DEHP in water according to OECD 433 guide line and sacrificed just after weaning. The weights of uterus and ovary were decreased by drinking of $1,330{\mu}g/L$ DEHP water. There was not adverse effects on either accumulated mating rate and mating rate depend on estrus stage, pregnancy duration, and sex ration at birth. However, the accumulated rate of successful delivery and litter size were significantly high at $1,330{\mu}g/L$ DEHP water. The number of epididymal sperm was significantly increased by drinking of $1,330{\mu}g/L$ DEHP water. In addition, the number of follicles (primary, secondary, tertiary) were more many than control at $1,330{\mu}g/L$ DEHP water drunk mother. Though further studies are needed to identify what are the mechanism of DEHP in folliculogenesis and spermatogenesis. From this study we firstly report the effect of low-dose chronic administration of DEHP with drinking could change the ovarian follicle population size and spermatogenesis rate. Put together, those finding is different from previous high-dose effects and suggest the physiological role of DEHP in gonads and uterus.

Studies on the Effects of Bisection Method and With and Without-Zona Pellucida of Bovine Embryos on In Vitro Developmental Rates (분할방법 및 투명대 부착 여부가 분할 초기배의 체외발생에 미치는 영향에 관한 연구)

  • 이종진;남윤이;김상근
    • Journal of Embryo Transfer
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    • v.12 no.1
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    • pp.21-27
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    • 1997
  • The studies were carried out to investigate the effects of bisection method and with and without-zona pellucida of embryos on in vitro developmental rate bisected embryos by micromanipulator, micropipette and pipetting. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes were cultured in TCM-199 mediurn containing 10 IU /ml의 PMSG(Sigma, USA), 10 IU /ml의 hCG, 1$\mu$g /ml의 $\beta$-estradiol(Sigma, USA) and 10% FCS for 24~48 hrs in incubator with 5% $CO_2$ in air at 38.5$^{\circ}C$ and then, matured oocytes were again cultured for 12~ 18 hrs with motile capacitated sperm by preincubation of heparin. Bisected embryos cultured for 1~5 days in 20% FCS + TCM-199 medium. Survival rate was defined as developmental rate on in vitro culture or FDA-test. The results are summarized as follows :1. The survival rates of bisected bovine embryos by micromanipulator and micropipett were 29.2% and 19.1%, respectively. The rates of non-bisection embryos(46.7%) were significantly higher than those of bisection embryos. 2. The in vitro developmental rates of bisected bovine embryos by micromanipulator, micropipett and pipetting method were 32.4%, 19.4% and 25.6%, respectively.3. The in vitro developmental rates of with and without-zona pellucida of bisected bovine embryos by raicromanipulator were 30.8% and 25.0%, respectively. The rates of nonbisection embryos(53.1%) were significantly higher than those of bisection embryos.

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Studies on the Effects of Co-culture of Cumulus Cell, Oviduct Epithelial Cell and Hormones and Freezing on !fl Vitro Developmental Rates of Bovine Embryos (소 수정란의 난구세포, 난관 상피세포, 호르몬과의 공배양 및 동결이 체외발생에 미치는 영향에 관한 연구)

  • 이종진;이명헌;김상근
    • Journal of Embryo Transfer
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    • v.12 no.1
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    • pp.27-36
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    • 1997
  • The studies were carried out to investigate the effects of co-culture with cumulus cells and oviduct epithelial cells on the in vitro fertilization and cleavage rate of bovine follicular cocytes and to determine the optimum thawing temperature and equilibration time on in vitro developmental rate of frozen bovine embryos. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes were cultured in TGM-199 medium containing 10 IU /ml의 PM SG, 10 IU /ml의 hCG, ip g/ml의 $\beta$-estradiol and 10% FCS for 24~48 hrs in incubator with 5% $CO_2$ in air at 38.5$^{\circ}C$. The bovine embryos following dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquld nitrogen and thawed in 3$0^{\circ}C$ water. Survival rate was defined as developmental rate on in vitro culture or FDA-test. The results are sunanarized as followes :1. The in vitro fertilization and in vitro developmental rates of bovine oocytes co-cultured with cumulus cells in TCM499 medium were 75.0~76.8% and 17.3~27.6%, respect-ively. And in-vitro fertilization rates of cumulus-enclosed oocytes(55.4%)were significantly(p<0.05) higher than cumulus-denuded oocytes (23.1%). 2. The in vitro fertilization and in vitro developmental rates of bovine oocytes co-cultured with l$\times$ l04cells /ml, 1 x l06cells /ml, lx l08cells /ml and 1 x l015cells /ml oviduct epithelial cells in TCM-199 medium were 74.5~77.8% and 15.7~21.20 respectively.3. The in-vitro fertilization and in vitro developmental rates of bovine oocytes cocultured in '1CM-199 media containing PMSG, hCG, PMSG+hCG. PMSG+$\beta$-estradiol, hCG+$\beta$-estradiol 0 to 40 hrs after insemination were 74.0~77.4% and l8.9~23.l%, re-spectiv ely.4.The survival rates of bovine embryos thawed after rapid freezing in the freezing medium containing a various concentration of sucrose added 1.5M and 2.OM glycerol,DMSO and propanediol were 23.5~31.4% and 20.6~34.l%, respectively. 5. The temperature thawed at 3$0^{\circ}C$ after rapid freezing of bovine embryos resulted in a significantly higher embryos survival rate than did at 2$0^{\circ}C$ and 35$^{\circ}C$.6. The equilibration time on the survival rates of bovine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time (10~20min.). (Key words : bovine embryos, co-culture, freezing, in vitro development)

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A Comparisons of Characteristics of Infants Born Prematurely According to Results of Denver II Screening Test (추후 덴버발달스크리닝 결과에 따른 미숙아의 특성 비교)

  • Bang, Kyung-Sook
    • Child Health Nursing Research
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    • v.12 no.3
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    • pp.398-404
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    • 2006
  • Purpose: To investigate the rate of questionable development in infants born prematurely and explore factors affecting developmental delays. Method: The participants were 46 infants born prematurely being seen in one of two urban health centers. A questionnaire and the HOME checklist were used to collect data, and the Korean Denver II developmental screening test was administered. Results: Of the participants 21.7% were classified as having questionable development. The only variable with a significant difference between the two groups was acceptance in the HOME checklist. Psychosocial factors such as mothers' burden, depression, family functioning, and social support were not significantly different between the two groups. Conclusion: An early developmental screening test for prematurely born infants is needed. Also, the childrearing environment was identified as a significant factor in infants' development. These findings suggest that HOME score might be useful for identifying infants at risk for developmental delays and interventions for these infants will probably be more effective if their mothers can provide a more appropriate social environment. Further studies are suggested with larger samples.

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Effects of Morphology, Reproductive Cycle, Incubation Time and Activation of Oocytes on Developmental Rate of Embryos Fertilized in vitro (난자의 형태, 번식주기, 배양시간 및 활성화 처리가 개 난자의 체외수정후 발생에 미치는 영향에 관한 연구)

  • 이동수;김상근
    • Journal of Embryo Transfer
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    • v.18 no.1
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    • pp.27-33
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    • 2003
  • The study was carried out to investigate the effects of morphology, reproductive cycle, incubation time and activation of oocytes in vitro maturation of canine oocytes and development of canine IVM/IVF embryos. The results were summarized as follows: 1. The developmental rates to 16 cells of fresh, salts and 4$^{\circ}C$-stored oocytes with and without cumulus cells were 14.3%, 5.0% and 7.5%, 2.8% and 5.7%, 0.0%, respectively. The rate of oocytes with cumulus cells(5.7%~14.3%) was higher than that of denuded oocytes(0.0%~5.0%). 2. The developmental rate to If cells of in vitro cultured oocytes recovered from ovaries collected at different stages of the reproductive cycle were 0.0%, 10.7%, 1.5%, respectively. 3. The developmental rate to 16 cells of fresh oocytes with cumulus cell cultured for 24, 32 and 48 hrs in $CO_2$ incubator were 0.0%, 5.3%, 11.8%, respectively. The rate of oocytes cultured for 48 hrs was higher than that oocytes cultured for 24 and 32 hrs. 4. The development to If cells treated activation and non-activation oocytes were 15.0%, 6.7%, respectively. The rate of oocytes treated activation was higher than that oocyte treat non-activation.

Study on the Factors Influencing Fertilization and Developmental Rate of In Vitro Cultured Cat Oocytes Recovered from Ovaries Collected at Different Stages of the Reproductive Cycle

  • Quan, J. H.;Kim, S. K.
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.257-257
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    • 2004
  • The study was carried out to investigate the effects of morphology, reproductive cycle, incubation time and activation of oocytes on in vitro maturation of cat oocytes and development of IVM/IVF embryos. 1. When recovered from ovaries collected at different stages of the reproductive cycle (inactive, follicular and luteal stage), the developmental rates of oocytes to GV and MI stage were 72.5% and 27.5%, 57.5% and 7.5%, 62.5% and 17.5%, respectively. (omitted)

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Analysis of Risk Factors in Children with Suspected Developmental Delays on the Denver Developmental Screening Test

  • Bang, Kyung-Sook
    • Child Health Nursing Research
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    • v.14 no.3
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    • pp.261-268
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    • 2008
  • Purpose: This study was conducted to estimate the prevalence of questionable development according to developmental screening testing for children, and to explore biological and environmental risk factors of developmental delays. Method: The study participants were 153 children under two years of age. A questionnaire and Home Observation for measurement of the Environment (HOME) inventory were used to collect data, and Korean Denver II was administered. Results: The overall prevalence of questionable development was 11.1%. Significant differences in biological factors between the normal development group and questionable development group were the rate of prematurity, twins, the educational level of mother and father's and the presence of an illness in the mother (p<.10). Maternal acceptance in HOME was significantly different between the two groups. The factors related to questionable development that were significant in the logistic regression analysis were prematurity (OR=3.56, p=.026), and maternal acceptance in HOME score (OR=.629, p=.028). Conclusion: Early developmental screening tests seem necessary for all children, especially for prematurely born children. Also, child rearing environments were identified as significant factors in the development of all children. These findings suggest that the HOME scores might be useful in identifying children at risk for developmental delays, and that interventions for these children will probably be more effective if their mothers are helped to provide a more appropriate social environment.

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