• Title/Summary/Keyword: Early development

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Functional Analyses of Centrosomal Proteins, Nek2 and NuMA in Development of Mouse Gametes and Early Embryos

  • Youn, Hong-Hee;Oh, Hwa-Soon;Lee, Kwang-Hee;Son, Chae-Ick;Lee, Sang-Ho
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.96-96
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    • 2003
  • Nek2 (NIMA-related protein) is a mammalian cell cycle-regulated kinase that involves in chromosome condensation and centrosome regulation and NuMA (nuclear mitotic apparatus protein) is involved in spindle assembly during a cell cycle. The cellular distribution and organization of the centrosomal components is completely unknown during fertilization and embryonic development. We examined distribution of two well-known centrosomal proteins, Nek2 and NuMA in mouse gametes and embryos to get an insight in the reorganization of centrosomal proteins during germ cell development and early fertilization. Spermatogenic cells, gametes, and embryos were analyzed with anti-Nek2 or -NuMA antibodies by immunological assay, RT-PCR, and overexpression through gene transfection. Mitotically or meiotically active spermatogenic cells were intensively stained with these antibodies in both centrosomes and cytoplasm, whereas the oocytes showed different staining patterns depending on the meiotic stages. During maturation, GV, GVBD, and MI stage were clearly stained with NuMA antibody in the nucleus or cytoplasm at MII. Also, Nek2 was detectable in cytoplasm as scattered spots or chromosome associated at MII. In early developmental embryo, NuMA was detected in nucleus of each blastomere, while Nek2 was detected in cytoplasm. In contrast to previously reported results, Nek2 and NuMA were detected in both decondensing head, and the centriole of demembranated and decondensed sperm or whole body of trypsin-treated sperm for Nek2. During meiotic progress in oocytes, transcripts levels were the highest in MI stage and then downregulated in MII. Also, it shows dramatically change in early developmental embryos, firstly, it was increased until 4 cell stage and reduced in 8 cell stage, and finally, transcript levels were upregulated until blastoscyst. This finding suggests that cnetrosomal component may play an important role in reorganizing of functional centrosome during fertilization process and subsequent development.

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Early Mouse Embryonic Development In Vitro by Co-culture with Bovine and Porcine Oviductal Epithelial Cells (소와 돼지의 난관 상피세포와의 공배양이 마우스 초기배의 체외발달에 미치는 영향)

  • Lee, S.;Hur, E. J.;Seok, H. B.
    • Journal of Embryo Transfer
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    • v.11 no.3
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    • pp.201-210
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    • 1996
  • This experiment was carried out to evaluate the effect of early mouse embryonic development in vitro by co-culture with bovine and porcine oviductal epithelial cells (BOEC and POEC). The 2-cell embryos were collected from the oviducts of the superovulated and mated cultured in D-PBS /15% FCS at 48 hours after hCG injection. The in vitro developmental rate of blastocyst formation in the embryos were examined under the fllowing treatments; 1) TCM 199 added 15% HCS, 2) Ham's F-10 added 15% HCS, 3) MediCult IVF medium, 4) TCM 199 added 15% HCS + BOEC, 5) TCM 199 added 15% HCS + POEC, 6) Ham's F40 added 15% HCS + BOEC, 7) Ham's F-10 added 15% HCS + POEC,8) MediCult IVF medium + BOEC, 9) MediCult IVF medium + POEC. For a comparative study of in vitro development for 96 hours after hCG injection, were cultured with oviductal epithelial cell and media only. The obtained results were 2-cell embryos developed to the blastocyst stage in TCM 199, Ham's F-10 and MediCult IVF medium at the rates of 84.4,83.2 and 81.6%. respectively. The higher developmental rates(91~97%) of blastocyst formation was appeared when the embryos were co-cultured with a monolayer of bovine or porcine oviductal epithelial cells in TCM 199 or Ham's F-10 and MediCult IVF media. No significant difference in developmental rates was shown between bovine and porcine oviductal epithelial cells but significant difference in co-culture system in comparison between media only system and co-cultures. In conclusions, oviductal epithelial cells, BOEC and POEC, when co-culture with mouse early embryos improved the rates of development, blastocyst and hatching. Therefore, it is suggested that co-culture system using oviductal epithelial cells improve early embryonic developtnent in mouse.

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Clinical efficacy and mechanism of probiotics in allergic diseases

  • Kim, Ha-Jung;Kim, Hyung Young;Lee, So-Yeon;Seo, Ju-Hee;Lee, Eun;Hong, Soo-Jong
    • Clinical and Experimental Pediatrics
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    • v.56 no.9
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    • pp.369-376
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    • 2013
  • A complex interplay between genetic and environmental factors partially contributes to the development of allergic diseases by affecting development during prenatal and early life. To explain the dramatic increase in the prevalence of allergic diseases, the hygiene hypothesis proposed that early exposure to infection prevented allergic diseases. The hygiene hypothesis has changed to the microbial hypothesis, in which exposure to microbes is closely linked to the development of the early immune system and allergic diseases. The intestinal flora may contribute to allergic disease through its substantial effect on mucosal immunity. Based on findings that exposure to microbial flora early in life can change the Th1/Th2 balance, thus favoring a Th1 cell response, probiotics may be beneficial in preventing allergic diseases. However, evidence from clinical and basic research to prove the efficacy of probiotics in preventing allergy is lacking. To date, studies have yielded inconsistent findings on the usefulness of probiotics in allergic diseases. It is difficult to demonstrate an exact effect of probiotics on asthma, allergic rhinitis, and food allergy because of study limitations, such as different first supplementation period, duration, different strains, short follow-up period, and host factors. However, many studies have demonstrated a significant clinical improvement in atopic dermatitis with the use of probiotics. An accurate understanding of the development of human immunity, intestinal barrier function, intestinal microbiota, and systemic immunity is required to comprehend the effects of probiotics on allergic diseases.

The Development of Early Mouse Embryos Depend on $Ca^{2+}$ Concentration (Ca$^{2+}$ 농도에 따른 생쥐 초기배의 발생)

  • Yang, Jung-Sook;Bae, In-Ha
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.3
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    • pp.275-282
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    • 2000
  • Objective: This study was to determine the effect of different concentration of calcium III medium on the preimplantational development of zygotes and early 2 cell embryos. Methods: Female mice of ICR strain ($5{\sim}8$ weeks old) were superovulated and mated with fertile males. Zygotes or early 2-cell embryos were collected by flushing the oviducts $31{\sim}32$ hours after hCG injection. The embryos were cultured in various concentrations of $Ca^{2+}$ in medium or with EDTA, EGTA and $Ni^{2+}$. Result and Conclusion: Treatment of high concentraion of $Ca^{2+}$ (3.42 mM $(2X){\sim}17.l$ mM (10X)) in medium didn't develop well compared to the control. Low concentrations of $Ca^{2+}$ (0.214mM $(1/8X){\sim}0.855$ mM (1/2X) were deterimental to development beyond 2-cell stage. EDTA, $Ca^{2+}$ chelating agent was treated with ranged concentrations of EDTA (0.014 $mM{\sim}0.107$ mM) to medium contaning 1.71 mM $Ca^{2+}$ showed beneficial effect to development to blastocyst compared to the control. EGTA, extracellular $Ca^{2+}$ chelator, was treated with ranged concentrations of EGTA ($0.014{\sim}0.107$ mM) to the medium contaning 1.71 mM $Ca^{2+}$. There is no significant difference with the control. $Ni^{2+}$ (50 ${\mu}M$), T-type $Ca^{2+}$-channel blocker was treated to medium contaning low concentration of $Ca^{2+}$. It overcame 2-cell block significantly. Rate of degenerated embryos decreased and developmental rate to morula and blastocyst increased more than low $Ca^{2+}$ concentration alone. Further studies are needed for the overcoming effect of 2-cell block by $Ni^{2+}$.

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Population changes and growth modeling of Salmonella enterica during alfalfa seed germination and early sprout development

  • Kim, Won-Il;Ryu, Sang Don;Kim, Se-Ri;Kim, Hyun-Ju;Lee, Seungdon;Kim, Jinwoo
    • Food Science and Biotechnology
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    • v.27 no.6
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    • pp.1865-1869
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    • 2018
  • This study examined the effects of alfalfa seed germination on growth of Salmonella enterica. We investigated the population changes of S. enterica during early sprout development. We found that the population density of S. enterica, which was inoculated on alfalfa seeds was increased during sprout development under all experimental temperatures, whereas a significant reduction was observed when S. enterica was inoculated on fully germinated sprouts. To establish a model for predicting S. enterica growth during alfalfa sprout development, the kinetic growth data under isothermal conditions were collected and evaluated based on Baranyi model as a primary model for growth data. To elucidate the influence of temperature on S. enterica growth rates, three secondary models were compared and found that the Arrhenius-type model was more suitable than others. We believe that our model can be utilized to predict S. enterica behavior in alfalfa sprout and to conduct microbial risk assessments.

A Study on the Participatory Housing Project Process in Strasbourg, France - Focusing on the Early Development Process of Ecologis - (프랑스 스트라스부르그(Strasbourg) 지역의 참여주택 프로젝트 과정 연구 - 에코로지스(Ecologis)의 초기 개발 단계를 중심으로 -)

  • Noh, Jun Seok
    • Journal of the Korean Institute of Rural Architecture
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    • v.26 no.1
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    • pp.33-40
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    • 2024
  • In France, the popularization of social rental housing development for regional balanced growth has encountered difficulties due to the increasing costs of housing development in the regions. As a new solution, since 2010, efforts have been made to promote participatory housing in the regions. As of 2023, Strasbourg has 284 households participating in the participatory housing projects. The Strasbourg area has many abandoned houses, and the land is owned by local governments. The local authorities are providing such lands for participatory housing and have established the "Eco-quartier" support association to efficiently support the project and assist in resident-led participatory housing development. This study analyzes the development characteristics of 26 completed participatory housing projects in the Strasbourg area and examines the initial development stages of the representative case, the "Ecologis" project. The success factors of the "Ecologis" project were the active administrative support of the local government, promotion for member recruitment, and the role of "Eco-quartier" in providing expert support teams. In the initial development stage, it identifies the distinctions from a similar concept in Korea, "community housing," and presents implications for the revitalization of participatory housing.

Morphological Analysis of Blind-Side Hypermelanosis of the Starry Flounder, Platichthys stellatus during Early Development

  • Kang, Duk-Young;Byun, Soon-Gyu;Myeong, Jeong-In;Kim, Hyo-Chan;Min, Byoung-Hwa
    • Development and Reproduction
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    • v.18 no.2
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    • pp.79-87
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    • 2014
  • In Pleuronectiformes, blind-side malpigmentation (hypermelanosis) is common in cultured flatfishes, and is economically important. To understand the mechanism of blind-side hypermelanosis in flatfishes, we examined when the malpigmentation initially occurred, and studied how the symptoms proceeded during early development of the starry flounder, Platichthys stellatus. To assess quantitative pattern changes of blind-side skin, we observed morphological development of the whole body from 22 (total length [TL] $10.0{\pm}0.2$ mm and body weight [BW] $8.8{\pm}0.57$ mg) to 110 days (TL $23.4{\pm}0.7$ mm, BW $193.6{\pm}23.3$ mg) after hatching (DAH), and also examined the malpigmented area rate of blind-side skin and the malpigmented fish ratios. The experimental animals were reared in fiberglass-reinforced plastic (FRP) tanks in water at a temperature of $18.9{\pm}1.9^{\circ}C$ and salinity of $32.6{\pm}0.6$ psu and were fed with rotifer and Artemia nauplii from 22 to 48 DAH, and with A. nauplii and commercial feed from 49 to 110 DAH. As results, the first staining patch seen by the naked eye was observed around the area between the anus and pelvic fin or caudal edge of the trunk at 80 DAH (TL $20.6{\pm}0.5$ mm, BW $112.5{\pm}8.8$ mg). The pigmented area and the pigmented fish ratios were significantly increased from 80 to 110 DAH. These results indicated that malpigmentation on the blind side of starry flounder was initially observed at about 2 cm in length and 100 mg in weight, and the pigmented domain on the blind-side skin was continually broadened by the differentiation of pigmented cells (melanophores and xanthophores) with growth.

Early Development of Parthenogenetically Activated Porcine Oocyte after In Vitro Maturation for Various Periods (난자성숙시간에 따른 처녀발생유기 돼지난자의 초기발생)

  • Kim, S. B.;Lee, H.;Byun, T. H.;Jeon, J. T.;Lee, S. H.;Song, H. B.
    • Journal of Embryo Transfer
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    • v.9 no.1
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    • pp.117-125
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    • 1994
  • In vitro development of parthenogenetic embryo was examined after ethanol treatment of follicular oocytes matured in vitro for 42, 48, 54 and 60h in the pig. The follicular oocytes were matured in TCM 199 containing 15% FCS and gonadotrophins in an atmosphere of 39 $^{\circ}C$ 5% $CO_2$. The cumulus-free oocytes were activated by 10% ethanol treatment in M2+4mg /ml BSA for 10 min. The ethanol-activated oocytes were washed and further cultured in TCM199+20%FCS containing granulosa cell monolayer. Maturation rates at 42, 48, 54 and 60h of IVM were 75.0, 86.5, 81.6 and 87.9%, respectively. Thus the oocytes maturated in vitro for longer periods did not improve nuclear maturation much. Pronuclear formation rates at 18h post-activation in ethanol-activated oocytes were 21.9, 25.0, 47.4 and 32.6%. The cytoplasmic maturation leading to pronuclear formation upon activation increased when the I VM period was extended from 42 to 54h. When the activated oocytes were cultured for 96~120h to analyse early development of the activated oocytes, the rates of embryonic development upto $\leq$ 5~8 cell were 5.3, 5.8, 12.0 and 11.7% among the cultured embryos. The result indicate that earlier development of activated porcine occyte is dependent on the duration of oocyte maturation, and that better development could be obtained from the oocyte matured for 54h.

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EARLY DEVELOPMENT OF THE TOOTH IN THE STAGED HUMAN EMBRYOS AND FETUSES (한국인 배자 및 태아에서 유치 발생의 조직학적 변화)

  • Lim, Hee-Sik;Park, Hyoung-Woo;Oh, Hyeon-Joo;Kim, Hee-Jin;Choi, Byung-Jai
    • Journal of the korean academy of Pediatric Dentistry
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    • v.25 no.2
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    • pp.383-399
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    • 1998
  • Tooth development is usually described in four stages such as bud stage, cap stage, bell stage and crown stage. Exact time of appearance of tooth primordia is different among reports, and up to now there is no timetable regarding initial tooth development. To understand the congenital malformations and other disorders of the orofacial region, there is a need to establish a standard timetable on early tooth development. Till now, studies on the tooth development were mainly on later fetuses, and only few reports on early stage. Also, there were no reports on the time when bud stage turns to cap stage, and cap stage to bell stage. In this study, external morphology of face and the early development of the tooth, and transition of bud stage to cap stage, cap stage to bell stage were studied using 27 staged human embryos and 9 serially sectioned human fetuses. The results are as follows: 1. Mandibular region was formed by union of both mandibular arch at stage 15, and maxillary region by union of maxillary arch, medial nasal prominence, and intermaxillary segment at stage 19. 2. Ectodermal thickening which represents the primordia of tooth appeared in mandibular region at stage 13, and maxillary region at stage 15. 3. Bud stage began from mandibular primary central incisor at stage 17, and maxillary primary central incisor at stage 18. And the sequence of appearance was in the mandibular primary lateral incisor at stage 19, maxillary primary lateral incisor at stage 20, mandibular primary canine at stage 22, maxillary primary canine and primary first molar at stage 23, madibular primary first molar and maxillary primary second molar at 9th week, and mandibular primary second molar at 10th week of development. 4. Cap stage began from the primary anterior teeth at 9th week, and primary second molar still had the characteristics of cap stage at 12th week of development. 5. Transition to bell stage started from the primary anterior teeth at 12th week, and primary second molar started at 16th week of development. 6. Trnasition to crown stage started from primary anterior teeth at 16th week, and primary second molar at 26th week of development.

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