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Systems for Production of Calves from Hanwoo IVM/IVF/IVC Blastocyst. IV. Direct Transfer of Vitrified and One-Step Diluted Hanwoo Blastocysts

  • 김은영;박세필;김덕임;이문걸;이종우;이금실;박세영;박은미;윤지연
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.73-73
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    • 2001
  • This study was to examine whether the vitrified, one-step diluted and direct transferred Hanwoo IVM/IVF/IVC blastocysts can be successfully survived in vivo and they were succeeded into the live birth. For vitrification, blastocysts were serially exposed in glycerol (G) or/and ethylene glycol (EG) mixtures [10% (v/v) G for 5 min, 10% G plus 20% EG (v/v) for 5 min, and 25% G plus 25% EG (v/v) for 30 sect] which is diluted in 10% FBS added D-PBS. Thawing of straw was carried out in air for 10 sec and then in water bath of $25^{\circ}C$ for 20 sec. One-step dilution within the straw was done in water bath of $25^{\circ}C$ for 1 min. Vitrified and one-step diluted embryos were directly transferred into 36 (natural or hormone induced synchronized) recipient cows in 6 areas of Kyungsang Buk-Do. Pregnancies were confirmed at first when recipient cows did not return to the subsequent estrus cycle, and later by manual palpation per rectum on day 45, 90 and then living calves were derived into parturition. Overall pregnancy was 33.3%(12/36), However, higher pregnancy was obtained when the recipients exhibited estrus one day earlier than the age of transferred embryos (53.3 vs 25.0-27.3%), irrespective of synchronization methods. Also, parous recipients became pregnant higher than nulliparous heifers, And, there were not different in pregnancy rates by the aspect of corpus luteum (CL) quality of recipients (good, 29.4; fair, 37.5; poor, 33.3%). One hundred eight of frozen-thawed Hanwoo blastocysts were directly transferred into 36 recipient cows. In 12 of pregnant cows, 3 cows were aborted and 9 cows were calved [single, 66.7% (6/9): twin, 33.3% (3/9)]. Total embryo implantation rate was 11.1% (12/108). However, 9 Hanwoo calves were lived. Therefore, these results demonstrate that direct transfer technique of vitrified and one-step diluted bovine blastocysts can be applied easily and effectively with the higher pregnancy rate on field trial without the equipment and embryological skills.

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Development of the Gonads Derived from Hetero-Sexually Transferred Primordial Germ Cells (PGCs) between Embryos in the Chicken

  • Furuta, H.;Yamaguchi, H.;Fujihara, N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.8
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    • pp.1188-1191
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    • 1999
  • Primordial germ cells (PGCs) of White Leghorn chicken embryos as a donor were transferred to Rhode Island Red chicken embryos as a recipient. At 48-50 h (stage 13-15) of incubation of fertilized eggs, donor PGCs, which were taken out from blood vessels of donor embryos, were injected into blood vessels of recipient embryos. Sex of the treated embryos was determined after the transfer of PGCs using remaining blood samples. In the present experiments, survival rate of the treated embryos was 33.3% for homo-sexual and 35.4% for hetero-sexual transfers of PGCs, respectively, when determined at 17 days of incubation. In this study, most of the treated embryos could not survive more than 18 days of incubation, though the reason for that was not clarified in the present work. The gonalds removed from embryos that died after 18 days of incubation and the organs from newly hatched chicks were examined for morphological and histological features. The gonads removed from the embryos with homo-sexual transfer of PGCs showed normal development in appearance. On the contrary, some (35.3%) of the embryos with hetero-sexual transfer of PGCs possessed abnormal gonads similar to ovotestis by histological observation. In cases where the gonads developed to be normal organs (64.7%) the sex of embryos was the same as recipient ones. The present results suggest that hetero-sexual transfer of the PGCs may bring about the possibility of development of the embryos bearing sexually different gametes, spermatogonia or oogonia.

Transformation Conditions of Bacillus subtilis by Streptomyces rimosus Plasmid DNA (Streptomyces rimosus Plasmid DNA에 의한 Bacillus subtilis의 형질전환 조건)

  • Hong, Yong-Ki;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.11 no.1
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    • pp.75-79
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    • 1983
  • To exploit a suitable vector and recipient strain for molecular cloning in Bacillus subtilis, oxytetracycline-resistant plasmic DNA has been prepared from Streptomyces rimosus by phenol-buffer extraction of lysozyme-lysed cells and introduced into B. subtilis KPM 60 [St $r^{R}$-mutant of RM 125 (leu A8, arg 15, hsm $M^{-10}$ , hsr $M^{-10}$ )] by transformation. Oxitetracycline-resistant plasmid was well transferred into B. subtilis KPM 60 with average frequency of 10$^{-4}$ per $\mu\textrm{g}$ of DNA. The highest frequency of plasmid transformation was obtained after 3 hours incubation of recipient cells in the growth medium and 30 to 60 minutes incubation in the competence medium, and then 20 minutes contact of DNA and host cells. Optimum pH for competence was 7.5, and optimum temperature for transformation was 2$0^{\circ}C$.>.

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Evaluation of the Home-based Rehabilitation Program Based on CBR Model Through In-depth Interviews With Visiting Physical Therapists

  • Lee, Minyoung;Chung, Jinjoo;Hong, Hye Jung;Yoon, Bum Chul
    • The Journal of Korean Physical Therapy
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    • v.27 no.1
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    • pp.68-77
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    • 2015
  • Purpose: This study was conducted in order to explore home visiting therapists' self-perceived role, problems, and supplement points of the home-based rehabilitation (HR) program based on the community-based rehabilitation (CBR) model. Methods: Four home visiting physical therapists, who conducted the HR program, participated in this study. After completion of the HR program, in-depth interviews were conducted using a semi-structured questionnaire for participants to explore their self-perceived role, problems, and supplement points of the HR program. Results: Participants regarded the role of the HR program as for "the linkage between recipient & society", "maintenance & improvement of recipients' physical function", and "education of recipient about the way of self rehabilitation". The problems and supplement points were derived from all phases of the HR program, including "human & material resources", "training program for human resources for HR service", "selection of recipient of HR service", "contents of the intervention for HR service", and "duration of HR service". Conclusion: These findings indicate that participants well recognized the intention of the HR program based on the CBR model, and suggest that high-quality human resources with rich expertise and experiences, a training program for HR service led by experts, and selection of recipients led by experts are necessary for an effective HR program. In addition, strategies and capacities specified to HR service should be identified, and applied to the training program for human resources for HR service. The results of this study could provide useful information when the government decides on the next guideline for home visiting health service.

Reconstruction of Soft Tissue Defects in the Finger using Arterialized Venous Free Flaps (유리 동맥화 정맥 피판을 이용한 수지 연부조직 결손의 재건)

  • Lee, Young-Keun;Woo, Sang-Hyun;Lee, Jun-Mo;Ahn, Hee-Chan;Cheon, Ho-Jun
    • Archives of Reconstructive Microsurgery
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    • v.19 no.1
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    • pp.21-28
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    • 2010
  • Purpose: To report the clinical results of the use of arterialized venous free flaps in reconstruction in soft tissue defects of the finger and to extend indications for the use of such flaps based on the clinical experiences of the authors. Materials and Methods: Eighteen patients who underwent arterialized venous free flaps for finger reconstruction, between May 2007 and July 2009 were reviewed retrospectively. The mean flap size was 4.7${\times}3.2$ cm. The donor site was the ipsilateral volar aspect of the distal forearm in all cases. There were 8 cases of venous skin flaps, 5 cases of neurocutaneous flaps, 4 cases of tendocutaneous flaps, 1 case of innervated tendocutaneous flap. The vascuality of recipient beds was good except in 4 cases (partial devascuality in 2, more than 50% avascuality (bone cement) in 2). Results: All flaps were survived. The mean number of included veins was 2.27 per flap. Mean static two-point discrimination was 10.5 mm in neurocutaneous flaps. In 3 of 5 cases where tendocutaneous flaps were used, active ROM at the PIP joint was 60 degrees, 30 degrees at the DIP joint and 40 degrees at the IP joint of thumb. There were no specific complications except partial necrosis in 3 cases. Conclusions: An arterialized venous free flap is a useful procedure for single-stage reconstruction in soft tissue or combined defect of the finger; we consider that this technique could be applied to fingers despite avascular recipient beds if the periphery of recipient bed vascularity is good.

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STUDIES ON PRODUCTION AND EFFICIENT UTILIZATION OF LIVESTOCK EMBRYOS BY IN VITRO FERTILIZATION AND MICROMANIPULATION IV. NUCLEAR TRANSPLANTATION AND ELECTROFUSION FOR CLONING IN BOVINE FOLLICULAR OOCYTES

  • Chung, Y.C.;Kim, C.K.;Song, X.X.;Yoon, J.T.;Choi, S.H.;Chung, Y.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.8 no.6
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    • pp.641-645
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    • 1995
  • This study was conducted to develop a method for production of nuclear transplant bovine embryos using in vitro-matured (IVM) oocytes and to examine the effect of different conditions of electrofusion on fusion rate and developmental capacity of donor nucleus transplanted to enucleated oocytes. Eight- to sixteen-cell embryos derived from oocytes matured and fertilized in vitro used as donor blastomeres and IVM oocytes were used as recipient oocytes. Oocytes were enucleated immediately after 23-24 h IVM and then reconstituted with a donor blastomere in two different micromanipulation media. Fusion rate and subsequent development of the reconstituted oocytes was compared under the different electric stimuli and recipient oocyte ages. Success rate of enucleation was significantly higher in TCM-199 medium containing FCS than in DPBS. The high fusion rate(75-94%) and development (6.4-14.8%) to morulae and blastocyst (M + B) were obtained from 0.6-0.75 kV/cm DC voltage, although total cleavage was not different among the electric pulses. Most optimal condition of electric stimulation for fusion and development was 1 DC voltage of 0.75 kV/cm, in which 80.5% of oocytes were fused, 80.0% and 31.7% of which was cleaved and developed to M + B, respectively. No M + B was obtained from 1.2 kV/cm DC voltage regardless of pulse frequency. Recipint oocyte age at electrofusion greatly affected the cleavage and subsequent development to M + B, showing high rate at 40-41 h oocyte maturation. These results suggest that a suitable condition of electrofusion for donor nuclei derived from IVF may be 1-2 DC pulses of 0.7 kV/cm for $70{\mu}sec$ and that processing of a transplanted nucleus in IVM oocytes may be affected by maturation age of recipient oocytes.

The anatomical study of internal mammary perforators (내유방혈관 관통지에 대한 해부학적 연구)

  • Lim, Sung Yoon;Song, Hyun Suk;Pae, Nam Suk;Park, Myong Chul
    • Archives of Plastic Surgery
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    • v.36 no.1
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    • pp.24-28
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    • 2009
  • Purpose: As a recipient vessel, internal mammary vessels have many advantages for microvascular reconstruction of the breast. But the approach is time consuming and results in large morbidities. However, the perforating branches of the internal mammary vessels can be used to minimize such demerits. The purpose of this cadaver study is to clarify the location and diameter of the perforating branches of internal mammary vessels and to prove they are safe and reliable recipient vessels. Methods: We studied 11 formalin - fixed cadavers and dissected their anterior chests bilaterally. The chests were exposed using midline presternal incisions. We dissected and found all perforators at subfascial planes under loupe magnification. The number, external diameter, and the distance from the midline were measured. Result: The mean external diameter of the arterial perforators was 1.32 mm and the mean external diameter of the venous perforators was 1.48 mm. The largest arterial and venous perforators were most frequently found in the second intercostal space. The mean distance from the midline to the perforator was evaluated; the artery averaged 1.95 cm and the vein averaged 2.08 cm. Conclusion: This study will be helpful when using the internal mammary perforating vessels as a recipient vessel during breast reconstruction.

Study on Nucleo-Cytoplasmic Interaction by Somatic Cell Nuclear Transfer in Bovine (소 체내포 핵이식에 의한 핵-세포질 상호작용에 관한 연구)

  • 정희태;최종엽;박춘근;김정익;민동미
    • Journal of Embryo Transfer
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    • v.15 no.1
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    • pp.23-31
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    • 2000
  • This study was conducted to investigate the effects of quiescent treatment of donor cells and activation treatment time of recipient cytoplasm on nuclear remodeling and in vitro development of somatic cell-cloned bovine embryos. Serum starved, confluent and nonquiescent cycling adult skin cells were teansferred into enucleated oocytes. Nuclear transfer oocytes were activated at 30 min, 1 and 2 hrs after electrofusion. Some nuclear transfer embryos(23% to 35%) extruded a polar body, which was not affected by quiescent treatment of donor cells and activiation time of recipient cytoplasm. About 68% of nuclear transfer embryos fused with a serum starved cells has a chromatin clump, but which was not different from embryos fused with confluent(51%) and nonquiescent(47%) cells. The proportion of embryos with a single chromatin clump was sightly increased when nuclear transfer embryos were activated within 30 min after fusion(69%) compared to those were activated at 1 and 2 hrs after fusion, but there was not significantly different. Development rates to the blastocyst stage were 8.6% and 15.9% when serum starved and confluent cells were transferred, which were higher than that of control group. Developmental rate to the blastocyst stage was higher in embryos were activated within 30 min after fusion (17.3%) compared to those of embryos were activated at 1 and 2 hrs after fusion (P<0.05). From the present result, it is suggested that quiescent treatment of donor cells and activation time of recipient cytoplasm can affect the in vitro development. Quiescent plasm activation within 30 min after fusion could increase the number of embryos with a normal chromation structure, which results in increased in vitro development.

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Allograft Immune Reaction of Kidney Transp lantation Part 2. Immunosuppression and Methods to Assess Alloimmunity (신이식 후 면역반응의 이해 2부 이식면역검사와 면역억제제)

  • Kang, Hee-Gyung
    • Childhood Kidney Diseases
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    • v.12 no.2
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    • pp.133-142
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    • 2008
  • For solid organ transplant, ABO blood type of donor and recipient should be compatible in principle. Recent improvement of immunosuppressant made HLA typing not so important while no-mismatch transplant still shows the longest graft survival. PRA(panel reactive antibody) test is to screen and identify recipients with HLA sensitization. When solid organ transplant is scheduled, cross-match test of donor cell and recipient serum should be performed and positive result of cross-match prohibits transplantation. Donor specific antibody(DSA) test can predict the severity of recipient immune reaction against donor organ. Today's mainstay of allograft immunosuppressant regimen is triple therapy of steroid, calcineurin inhibitor(cyclosporine, tacrolimus), azathioprine or mycophenolate mofetil(MMF). Antibody induction using Thymoglobulin or anti-IL-2 receptor antibody(basiliximab or daclizumab) is frequently practiced as well.

Research on the Effects of a Project Manager's Competency on a Consultant's Ethical Attitude and the Consulting Achievement with the Regulation Effects of CEO Support (CEO지원을 조절효과로 프로젝트 매니저 역량이 컨설턴트의 윤리적 태도와 컨설팅 성과에 미치는 영향에 관한 연구)

  • Shin, Dong-Ju;You, Yen-Yoo
    • Journal of Digital Convergence
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    • v.12 no.5
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    • pp.191-201
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    • 2014
  • The purpose of this research is to verify the importance of the relationship between the competency of the project manager of a consulting recipient company that can significantly influence the consulting achievement, the ethical attitude of the consultant, and the consulting project achievement. The results of this research are summarized as follows. The results of this study verified that the competency of the project manager from the consulting recipient company contributed to the ethical attitude of the consultant, the ethical attitude of the consultant positively affected the consulting achievement, and the support of the CEO of the consulting recipient company helped regulate the relationship between the competency of the project manager and the consulting achievement.