• Title/Summary/Keyword: Reproductive capacity

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Derivation of Neural Precursor Cells from Human Embryonic Stem Cells

  • Kim Sehee;Hong Ji Young;Joo So Yeon;Kim Jae Hwan;Moon Shin Yong;Yoon Hyun Soo;Kim Doo Han;Chung Hyung Min;Choi Seong-Jun
    • Reproductive and Developmental Biology
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    • v.28 no.4
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    • pp.247-252
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    • 2004
  • Human embryonic stem (ES) cells are derived from the inner cell mass of the preimplantation embryo. Human ES cells have the capacity to differentiate into various types of cells in the body. Human ES cells are indefinite source of cells for cell therapy in various degenerative disorders including neuronal disorders. Directed differentiation of human ES cells is a prerequisite for their clinical application. The objective of this study is to develop the culture condition for the derivation of neural precursor cells from human ES cells. Neural precursor cells were derived from human ES cells in a stepwise culture condition. Neural precursor cells in the form of neural rosette structures developed into neurospheres when cultured in suspension. Suspension culture of neurospheres has been maintained over 4 months. Expressions of nestin, soxl, sox2, pax3 and pax6 transcripts were upregulated during differentiation into neural precursor cells by RT-PCR analysis. In contrast, expression of oct4 was dramatically downregulated in neural precursor cells. Immunocytochemical analyses of neural precursor cells demonstrated expression of nestin and SOX1. When induced to differentiate on an adhesive substrate, neuro-spheres were able to differentiate into three lineages of neural systems, including neurons, astrocytes and oligo-dendrocytes. Transcripts of sox1 and pax6 were downregulated during differentiation of neural precursor cells into neurons. In contrast, expression of map2ab was elevated in the differentiated cells, relative to those in neural precursor cells. Neurons derived from neural precursor cells expressed NCAM, Tuj1, MAP2ab, NeuN and NF200 in immunocytochemical analyses. Presence of astrocytes was confirmed by expression of GFAP immuno-cytochemically. Oligodendrocytes were also observed by positive immuno-reactivities against oligodendrocyte marker O1. Results of this study demonstrate that a stepwise culture condition is developed for the derivation of neural precursor cells from human ES cells.

Effects of Exogenous Insulin-like Growth Factor 2 on Neural Differentiation of Parthenogenetic Murine Embryonic Stem Cells

  • Choi, Young-Ju;Park, Sang-Kyu;Kang, Ho-In;Roh, Sang-Ho
    • Reproductive and Developmental Biology
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    • v.36 no.1
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    • pp.33-37
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    • 2012
  • Differential capacity of the parthenogenetic embryonic stem cells (PESCs) is still under controversy and the mechanisms of its neural induction are yet poorly understood. Here we demonstrated neural lineage induction of PESCs by addition of insulin-like growth factor-2 (Igf2), which is an important factor for embryo organ development and a paternally expressed imprinting gene. Murine PESCs were aggregated to embryoid bodies (EBs) by suspension culture under the leukemia inhibitory factor-free condition for 4 days. To test the effect of exogenous Igf2, 30 ng/ml of Igf2 was supplemented to EBs induction medium. Then neural induction was carried out with serum-free medium containing insulin, transferrin, selenium, and fibronectin complex (ITSFn) for 12 days. Normal murine embryonic stem cells derived from fertilized embryos (ESCs) were used as the control group. Neural potential of differentiated PESCs and ESCs were analyzed by immunofluorescent labeling and real-time PCR assay (Nestin, neural progenitor marker; Tuj1, neuronal cell marker; GFAP, glial cell marker). The differentiated cells from both ESC and PESC showed heterogeneous population of Nestin, Tuj1, and GFAP positive cells. In terms of the level of gene expression, PESC showed 4 times higher level of GFAP expression than ESCs. After exposure to Igf2, the expression level of GFAP decreased both in derivatives of PESCs and ESCs. Interestingly, the expression level of $Tuj1$ increased only in ESCs, not in PESCs. The results show that IGF2 is a positive effector for suppressing over-expressed glial differentiation during neural induction of PESCs and for promoting neuronal differentiation of ESCs, while exogenous Igf2 could not accelerate the neuronal differentiation of PESCs. Although exogenous Igf2 promotes neuronal differentiation of normal ESCs, expression of endogenous $Igf2$ may be critical for initiating neuronal differentiation of pluripotent stem cells. The findings may contribute to understanding of the relationship between imprinting mechanism and neural differentiation and its application to neural tissue repair in the future.

Bionomics of Caloglyphus sp.(Acarina : Acaridae) (가루응애류 Caloglyphus sp.의 생태에 관한 연구)

  • Park, Ji-Doo;Kim, Jong-Kuk
    • Journal of Forest and Environmental Science
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    • v.13 no.1
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    • pp.51-56
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    • 1997
  • Developmental characteristics and reproductive ability of Caloglyphus sp. were examined under the constant temperature conditions. This mites has five stages ; egg, larva, protonymph, deutonymph and adult. All active instars preyed on larvae of pine gall midge, Thecodiplosis japonensis Uchida et Inouye. The developmental period from the egg to the adult decreased with temperature increase at a range between $15^{\circ}C$ and $30^{\circ}C$. The threshold of temperature and the thermal constant for the development from the egg to the adult were $8.2^{\circ}C$ and 122.0 day-degrees, respectively. At $25^{\circ}C$, survival rate from egg to deutonymph was estimated as 66.4%, and longevity of the adult was 12.3 days for male and 10.0 days for female. After a preoviposition period of 1 and 2 days female laid an average of 360.6 eggs each, almost all during the first half of their life time. The value of the net reproduction(Ro), the mean length of a generation(T) and the intrinsic rate of natural increase($r_m$) were calculated as 101.1, 9.3 days and 0.494 per female per day, respectively.

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Comparison of Gene Expression Levels of Porcine Satellite Cells from Postnatal Muscle Tissue during Differentiation

  • Jeong, Jin Young;Kim, Jang Mi;Rajesh, Ramanna Valmiki;Suresh, Sekar;Jang, Gul Won;Lee, Kyung-Tai;Kim, Tae Hun;Park, Mina;Jeong, Hak Jae;Kim, Kyung Woon;Cho, Yong Min;Lee, Hyun-Jeong
    • Reproductive and Developmental Biology
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    • v.37 no.4
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    • pp.219-224
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    • 2013
  • Muscular satellite cell (SC), which is stem cell of postnatal pig, is an important for study of differentiation into adipogenesis, myogenesis, and osteoblastogenesis. In this study, we isolated and examined from pig muscle tissue to determine capacity in proliferate, differentiate, and expression of various genes. Porcine satellite cells (PSC) were isolated from semimembranosus (SM) muscles of 90~100 days old pigs according to standard conditions. The cell proliferation increased in multi-potent cell by Masson's, oil red O, and Alizarin red staining respectively. We performed the expression levels of differentiation related genes using real-time PCR. We found that the differentiation into adipocyte increased expression levels of both fatty acid binding protein 4 (FABP4) and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) genes (p<0.01). Myocyte increased the expression levels of the myosin heavy chain (MHC), myogenic factor 5 (Myf5), myogenic regulatory factor (MyoD), and Myogenic factor 4 (myogenin) (p<0.01). Osteoblast increased the expression levels of alkaline phosphatase (ALP) (p<0.01). Finally, porcine satellite cells were induced to differentiate towards adipogenic, myogenic, and osteoblastogenic lineages. Our results suggest that muscle satellite cell in porcine may influence cell fate. Understanding the progression of PSC may lead to improved strategies for augmenting meat quality.

Dose- Response Curves of Mouse Jejunal Crypt Cells by Multifractionated Irradiation (다분할조사에 의한 마우스공장소낭선 세포의 선량반응곡선)

  • Hong, Seong-Eon;Ahn, Chi-Yul
    • Radiation Oncology Journal
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    • v.4 no.2
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    • pp.89-97
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    • 1986
  • Using as assay for jejunal crypt stem cell survival, dose-response curves for the reproductive capacity of crypt stem cells of mouse jejunum exposed to multifractionated gamma-ray irradiation (single, 2, 3, 4, 5, 8, 10, 12, and 16 fractions) were analyzed and single-dose survival curve of these cells was constructed. The following conclusion were drawn: 1) Survival curves for higher numbers of dose fractions were displaced to higher dose, and characterized by increasingly shallower slopes. 2) The single-dose survival curve had broad shoulder, Dq=460 cGy, remaining near-exponential over initial dose range 0 to 300 cGy, with initial slope 1Do=474 cGy. 3) At fractionated dose En the range of 180 to 450 cGy, the average recovered dose per fraction interval was approximately $50\%$ of the dose per fraction. 4) The value of $\alpha/\beta$ ratio by using of linear regression analysis for the reciprocal dose plots was 8.3 Gy which lied in the range of 6-14 Gy for early-reacting tissues. 5) The linear-quadratic model for dose-response formula offers valid approximations for at 1 doses to be used in radiotherapy, only two parameters to be determined, and considerable convenience in practical applications.

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Evaluation of Four Plant Species as Potential Banker Plants to Support Predatory Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) in Cucumber Plant Systems (시설오이에서 미끌애꽃노린재의 천적유지식물로서 4종 식물의 평가)

  • YongSeok Choi;Gun-Woo Lee;Gyung-Ju Lee;Han-Jung Na;InSu Hwang
    • Korean journal of applied entomology
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    • v.63 no.1
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    • pp.25-32
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    • 2024
  • Frankliniella occidentalis and Thrips palmi are economically important pests of cucumber. The chemicals used to control them can be effective; however, they should not be used frequently on cucumbers that are consumed raw. Banker plants were selected to increase the control efficiency of Orius laevigatus, a natural enemy that can replace the chemicals. The hatching rate, emergence rate, nymph developmental period, survival rate, and reproductive capacity (fecundity and oviposition period) of O. laevigatus for four plants (moss rose, basil, broad bean, and cucumber) were investigated. The density of O. laevigatus on two selected banker plants was investigated in a cucumber greenhouse. The hatching rate of O. laevigatus eggs was highest at 92% in moss rose; however, there was no significant difference in survival rates between moss rose and basil. The fecundity and oviposition period of O. laevigatus were better in moss rose than in basil, with no significant difference between them. The flowering period of basil was longer than that of moss rose, from April to September. Therefore, basil has potential value as a banker plant for O. laevigatus and is expected to increase the biological control effect of O. laevigatus.

Effects of Energy Substrates in Culture Media on Developmental Capacity of Mouse Embryos (배양액에 첨가하는 에너지원이 생쥐 배 발생 능력에 미치는 영향)

  • Park, Kee-Sang;Lee, Hyun-Jung;Park, Sung-Baek;Kim, Ji-Chul;Lee, Taek-Hoo;Chun, Sang-Sik
    • Reproductive and Developmental Biology
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    • v.31 no.1
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    • pp.35-41
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    • 2007
  • This study was conducted to examine the effects of energy substrates in different conoentration of carbohydrates in the human oviduct and uterus on the in vitro development of mouse 2-cell embryos. Two-cell embryos were collected from ICR female mice at $46{\sim}50\;hr$ after 5 IU hCG injection and cultured in three different media [control: 0 mM, Guoup A: glucose (G) 0.5 mM + pyruvate (P) 0.32 mM + lactate (L) 10.5 mM, Group B: G 3.15 mM + P 0.1 mM + L 5.83 mM] for 72 hr. Rates of morula formation of group A (72.3%) and B (56.6%) were significantly higher higher (p<0.05) than that of control (34.9%) at 24 hr. However, blastocyst rate was significantly higher (p<0.05) in control (51.8%) than group A (39.8%) and B (28.9%) at hr. At 72 hr, no differences were found in the number of zona-intact, zona-escape and total blastocysts among groups. Mean and ICM cell numbers were significantly higher (p<0.05) in group A (78.0, 13.4) and B (64.4, 11.8) than control (53.1, 5.7), respectively, The percent of ICM were significantly higher (p<0.05) in group A (22.9%) and B (23.7%) than control (14.2%). No differences were round in the TE cell numbers ($34.1{\sim}45.1$). The ICM:TE ratio was significantly higher $34.1{\sim}45.1$ in control (1:6.0) than group A (1:3.4) and B (1:3.4). This study shows that energy substrates added to culture media especially, the oviductal level of carbohydrates increase the developmental capacity of 2-cell mouse embryos.

Studies on the Flowering and Maturity in Sesame (Sesamum indicum L.) III. Growth of Capsule and Grain by Different Plant Types (참깨의 개화.등숙에 관한 연구 III. 참깨 초형별 삭과 및 종실의 발육)

  • Kang, Chul-Whan;Lee, Jung-Il;Son, Eung-Ryong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.30 no.2
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    • pp.158-164
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    • 1985
  • The objective of the study was to investigate growth pattern of capsule and grain to improve grain filling during the grain filling period in sesame. Growth patterns of capsule and grain from anthesis to maturity were measured and compared by different plant types. Growth of capsule length started to grow just after anthesis and recorded maximum point at 35 days after flowering. and then decreased gradually. Growth of higher part capsule was worse than lower and middle capsules. Capsule growth of 2 carpels 4 loculi type showed better than 4 carpels 8 loculi type and BTB (branch, 3 capsules, 2 carpels, 4 loculi) type showed good growth due to its small reduction of higher part capsule length compared to those of lower and middle parts. The order of growth of capsule length were considered to be center capsule in main stem> center capsule in branch> side capsule in main stem> side capsule in branch. Growth of capsule width also showed maximum at 35 days after anthesis and then reduced. The order of growth of capsule width were lower part> middle part> higher part. Higher part capsule width of 3 capsules 4 carpels 8 loculi type showed serious decrease at late reproductive growth stage same as those of capsule length. Fresh one thousand grain weight showed peak at 35 days after anthesis and then reduced. The order of grain growth were appeared as lower part> middle part> higher part. Growth of fresh one thousand grain weight of branch and side capsule were lower than those of main stem center capsule, and 4 carpels 8 loculi type was deeply decreased at late flowering time in higher part such as the growth of capsule length and width. BTB (branch, 3,2/4) type didn't show much decrease in higher part fresh grain weight compared to those of other plant types in spite of its profitable character of lots of sink capacity. BTB type appeared to be ideal for improving grain filling and yield productivity in growing sesame in Korea.

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A Preliminary Epidemiologic Study on Korean Veterans Exposed to Herbicides in Vietnam War (파월국군장병의 고엽제 위해에 관한 예비적 역학조사)

  • Kim, Joung-Soon;Lee, Hyun-Sul;Lee, Hong-Bok;Lee, Won-Young;Park, Young-Joo;Kim, Sung-Soo
    • Journal of Preventive Medicine and Public Health
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    • v.27 no.4 s.48
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    • pp.711-734
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    • 1994
  • Among chemical agents in herbicides, dioxin (2, 3, 7, 8-tetrachloro dibenzo-$\rho$-dioxin : TCDD), a chemical contaminant in herbicides sprayed during the Vietnam War has been known to be the major agent causing toxic effects. Approximately 320,000 korean soldiers participated the Vietnam War from 1964 to 1974. Although the potential hazards of the herbicides among Korean veterans exposed were implicated, the problem had not been a public issue until 1991 when Korean veterans were informed U.S. companies, the herbicides manufacturer payed fund, from which a trust fund for New Zealand and Australian Class members were established in 1985. After a series of appeals and demonstration by the Korean Veterans demanding medical care and compensation for their serious health damages, a bill of medical care and compensation for herbicides victims was promulgated in March 1993 and become effective from May 1993, This study was carried out with two major objectives : the first to understand the health problems caused from the herbicides by reviewing literatures published, and the second to examine the nature and extent of health impacts among Korean veterans exposed and to develop valid study methods for the major study by interviewing and reviewing records on a part of veterans (638 persons) registered and completed medical examination in Seoul Veterans Administration Hospital from June to October 1993. The results obtained are as followings: 1. The literature review of 107 papers revealed that 1) Dioxin is teratogenic, carcinogenic and affects almost all organs including nervous, endocrine, and reproductive systems in animal experiments. 2) The diseases showing evidence of causal association were Hodgkin's disease, non-Hodgkin's disease, lung cancer, lymphoma, soft tissue sarcoma, chloroacne and polyneuropathy when judged on the basis of consistency in study results and biological plausibility. 2. Interview and medical record review study on 638 veterans, though limited validity owing to lack of control group, crude estimates of dioxin exposure levels (no biomarkers measurable), and uncertainty of diagnosis, showed that: 1) Most of the study subject's were in their 40's of age and had been dispatched to Vietnam during the period from $1965{\sim}1970$ around one year. 2) Most frequently complained symptoms in medical examination were motor weakness (32%), sensory abnormalities in extremities (23%), skin diseases (22%), and pain in extremities (20%) whereas in Interview they were more frequent in order of skin problem (44%), motor weakness (38%), sensory abnormalities and pain in extremities(17% and 19% each). Kappa indices on the same category of complaints between two sources of information were variable and relatively low. 3) On medical examination, only a part of the 638 subjects had initial impression (442 pts) and final diagnosis (218 pts) suggesting decision making on diagnosis appeared to be difficult even with all available modern medical technologies: in initial impression disorders from peripheral and central neuropathy were predominant whereas in final diagnosis various types of skin disorder were most frequent 4) When dose-response relationship between several conditions (from questionnaire) and arbitrary exposure scores were examined by CMH linear trend test, spontaneous abortion, sexual problems and health problem of offsprings showed statistically significant linear trends. However, pregnancy, accident and suicidal attempts did not show any relationship in this study capacity. 5) Among complaints, psychosis and neurosis (anxiety, phobia) in interview study, and memory disorder and psychosis in medical record study revealed linear trend. 6) Skin disorder was the only condition showing linear trend in initial impression and none in final diagnosis on medical examination. Even though objective to select out dioxin-related disease or group of diseases from this study was not achieved the research experiences provided firm basis for developing various methodological approaches. 3. From this preliminary study we concluded that a larger scale major epidemiologic study on health impacts of herbicides among Korean veterans exposed is not only Indispensible but also well designed study with more valid exposure information and diagnosis may be able to establish causal relationship between certain groups of diseases and exposure to the herbicides among Korean veterans.

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Pathogenicity bioassay of entomopathogenic nematode, Steinernema carpocapsae GSN1 strain (Nematoda: Steinernematidae) on the shiitake fungus moth, Morophagoides moriutii (Lepidoptera: Tineidae) (표고버섯좀나방에 대한 곤충병원성 선충의 병원성 검정)

  • Kim, Hyeong Hwan;Kim, Dong Hwan;Cho, Myung Rae;Yang, Chang Yeol;Kang, Taek Jun;Jeon, Sung Wook;Song, Jin Sun;Jung, Young Hak;Park, Hae Woong;Park, Chung Gyoo;Choo, Ho Yul
    • Journal of Mushroom
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    • v.12 no.4
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    • pp.357-362
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    • 2014
  • Pathogenicity of Steinernema carpocapsae GSN1 strain was evaluated against Morophagoides moriutii larvae. In Petridish tests, insect mortality by the nematode was dose dependent, which increased with dose from 5 to 160 infective juveniles(IJs)/larva. Pathogenicity against fourth-instar larvae was higher than the rate of corresponding second- and third-instar larvae, showing 100% insect mortality with the dose of 40 IJs/$4^{th}$ instar larva and 80 IJs/$2^{nd}$ or $3^{rd}$ instar larvae. Lethal concentration values at 50% ($LC_{50}$) of S. carpocapsae GSN1 strain were 4.2 IJs/$2^{nd}$ instar larva; 8.5 IJs/$3^{rd}$ instar larva; and 2.3 IJs/$4^{th}$ instar larva, respectively. The number of nematodes established in M. moriutii larvae after infection increased in the increment of dose and insect developmental stage. The highest number of nematodes was harvested from fourth instar larvae of M. moriutii at a dose of 160 IJs per larva, showing 22.5 nematodes per insect larva. Nematode reproductive capacity was related to insect developmental stage, showing 6,335 IJs/$2^{nd}$ instar larva, 21,660 IJs/$3^{rd}$ instar larvae, and 88,700 IJs/$4^{th}$ instar larvae.