• Korean Journal of Poultry Science
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• v.28 no.2
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• pp.135-142
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• 2001

A novel sterategy has been established to determine the origin of the Primordial Germ Cells (PGCs) in avian embryos directly and the developmental fate of the PGCs for the application to Poultry biotechnology. Cells were removed from 1) the centre of area pellucida, 2) the outer of area pellucida and 3) the area opaca of the stage X blastoderm (Eyal-Giladi & Kochav, 1976). When the cells were removed from the centre of area pellucida, the mean number of circulating PGCs in blood was significantly decreased in the embryo at stage 15 (Hamburger & Hamilton, 1951) as compared to intact embryos. When the cells were replenished with donor cells, no reduction in the PGCs number was observed. The removal of cells at the outer of area pellucida or at the area opaca had no effect on the number of PGCs. In case, another set of the manipulated embryos were cultured ex vivo to the hatching and reared to the sexual maturity, the absence of germ cells and degeneration of seminiferous tubules was observed in resulting chickens derived from the blastoderm in which the cells were removed from the centre of the area pellucida. It was concluded that the avian Primordial Germ cells are originated at the center of area pellucida. Developmental ability of the cells to differentiate into somatic cells and germ cells in chimeras were analyzed. Somatic chimerism was detected as black feather attributed from donor cells. Molecular identification by use of female - specific DNA was performed. It was confirmed that the donor cells could be differentiated into chimeric body and erythrocytes. Donor cells retained the ability to differentiate into germline in chimeric gonads. More than 70% of the generated chimeras transmitted donor derived gametes to their offspring indicating that the cells at the center of area pellucida had the high ability to differentiate into germ cells. A molecular technique to identify germline chimerism has been developed by use of gene scan analysis. Strain specific DNA fragments were amplified by the method. It would be greatly contributed for the detection of germline chimerism. Mixed- sex chimeras which contained both male and female cells were produced to investigate the developmental fate of male and female cells in ovary and testes. The sex combinations of donor and recipient of the resulting chimeras were following 4 pairs; (1) chimeras (ZZ/ZZ) produced by a male donor (ZZ) and a male recipient (ZZ), (2) chimeras (ZW/ZW) produced by a female donor (ZW) and a female recipient (ZW), (3) chimeras (ZZ/ZW) Produce by a male donor (ZZ) and a female recipient (ZW), (4) chimeras (ZW/ZZ) produced by a female donor (ZW) and a male recipient (ZZ). It was found that genetically male avian germ cells could differentiate into functional ova and that genetically female germ cells can differentiate into functional spermatozoa in the gonad of the mixed- sex chimeras. An ability for introduction of exogenous DNA into the PGCs from stage X blastoderms were analyzed. Two reporter genes, SV-$\beta$gal and RSV-GFP, were introduced into the PGCs. Expression of bacterial/gal was improved by complexing DNA with liposome detectedcc in 75% of embryos at 3 days embryos. At the embryos incubated for 1 day, expression of the GFP was observed all the embryos. At day 3 of incubation, GFP was detected in about 70% of the manipulated embryos. In case of GFP, expression of the transgene was detected in 30 %e of the manipulated embryos. These results suggested that the cells is one of the most promising vectors for transgenesis. The established strategy should be very powerfull for application to poultry biotechnology.

• Geophysics and Geophysical Exploration
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• v.10 no.4
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• pp.383-392
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• 2007

Karstic features and mining-related cavities not only lead to severe restrictions in land utilizations, but also constitute serious concern about geohazard and groundwater contamination. A microgravity survey was applied for detecting, mapping and monitoring karstic cavities in the test site at Muan prepared by KIGAM. The gravity data were collected using an AutoGrav CG-3 gravimeter at about 800 stations by 5 m interval along paddy paths. The density distribution beneath the profiles was drawn by two dimensional inversion based on the minimum support stabilizing functional, which generated better focused images of density discontinuities. We also imaged three dimensional density distribution by growing body inversion with solution from Euler deconvolution as a priori information. The density image showed that the cavities were dissolved, enlarged and connected into a cavity network system, which was supported by drill hole logs. A time-lapse microgravity was executed on the road in the test site for monitoring the change of the subsurface density distribution before and after grouting. The data were adjusted for reducing the effects due to the different condition of each survey, and inverted to density distributions. They show the change of density structure during the lapsed time, which implies the effects of grouting. This case history at the Muan test site showed that the microgravity with accuracy and precision of ${\mu}Gal$ is an effective and practical tool for detecting, mapping and monitoring the subsurface cavities.

• Microbiology and Biotechnology Letters
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• v.39 no.4
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• pp.337-344
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• 2011

Integrase MJ1 from the bacteriophage ${\Phi}FC1$ carries out recombination between two DNA sequences (the phage attachment site, attP and the bacterial attachment site, attB) in NIH3T3 mouse cells. In this study, the integration vector containing attP, attB and the integrase gene MJ, was constructed. The integration mediated by integrase MJ1 in Escherichia coli led to excision of LacZ. Therefore, the frequency of integration was measured by the counting of the white colony, which is detectable on X-Gal plates. The extrachromosomal integration in NIH3T3 mouse cells was monitored by the expression of the green fluorescent protein (GFP) as a reporter. To demonstrate integration mediated integrase MJ1 in NIH3T3 cells, vectors containing attP and attB were co-transfected into NIH3T3 cells. The integration was confirmed by fluorescent microscopy. The expression of GFP was induced in NIH3T3 cells expressing MJ1 without accessory factors. By contrast, the excision mediated by the MJ1 between attR and attL had no effect on the expression of GFP. These results suggest that integrase MJ1 may enable a variety of genomic modifications for research and therapeutic purposes in higher living cells.

• Journal of Chest Surgery
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• v.30 no.9
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• pp.920-926
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• 1997

Remarkable effect of pain relief and prevention of the postoperative Complications after thoracotomy has been achieved by continuous intravenous analgesia. This study was carried out with thirty patients who underwent posterolateral thoraco tony. The patients were divided into three groups: Group I(n= 10), the patients with intermittent intramuscular analgesia(piroxicam 20 mg), Group II(n=10), the patients with continuous epidural analgesia(0.5% bupivacaine 30m1 + normal saline 30 ml + morphine 10 mg), and Group III(n= 10) the patients with controlled intravenous infusion of analgesics(fentanyl 2500 mfg +normal saline 10 ml). The results w re as follows; 1) There were no significant changes of vital signs, between groups. 2) Tidal volume and FVC were significantly improved in the group II and III compared with the group I during the first postoperative day. 3) A significant reduction of immediate post-thoracotomy pain was achieved in the group II and III compared with the group I. 4) The limitation of motion in the operative side was less in the group II and III compared with the group I. 5) A signi(icant reduction of the postoperative analgegics consumption was noticed in group II and III. 6) Significant complications were not occured during follow-up period in all groups.

• Radiation Oncology Journal
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• v.17 no.4
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• pp.299-306
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• 1999

Purpose : The human genetic disorder ataxia-telangiectasia (AT) is a multisystem disease characterized by extreme radiosensitivity. The recent identification of the gene mutated in AT, ATM, and the demonstration that it encodes a homologous domain of phosphatidylinositol 3-kinase (PI3-K), the catalytic subunit of an enzyme involved in transmitting signals from the cell surface to the nucleus, provide support for a role of this gene in signal transduction. Although ionizing radiation was known to induce c-fos transcription, nothing is known about how ATM or PKCI mediated signal transduction pathway modulates the c-fos gene transcription and gene expression. Here we have studied the effect of PKCI on radiation sensitivity and c-fos transcription in normal and AT cells. Materials and Methods: Normal (LM217) and AT (AT5BIVA) cells were transfected with PKCI expression plasmid and the overexpression and integration of PKCI was evaluated by northern blotting and polymerase chain reaction, respectively. 5 Gy of radiation was exposed to LM and AT cells transfected with PKCI expression plasmid and cells were harvested 48 hours after radiation and investigated apoptosis with TUNEL method. The c-fos transcription activity was studied by performing CAT assay of reporter gene after transfection of c-fos CAT plasmid into AT and LM cells. Results: Our results demonstrate for the first time a role of PKCI on the radiation sensitivity and c-fos expression in LM and AT cells. PKCI increased radiation induced apoptosis in LM cells but reduced apoptosis in AT cells. The basal c-fos transcription activity is 70 times lower in AT cells than that in LM cells. The c-fos transcription activity was repressed by overexpression of PKCI in LM cells but not in AT cells. After induction of c-fos by Ras protein, overexpression of PKCI repressed c-fos transcription in LM cells but not in AT cells Conclusion: Overexpression of PKCI increased radiation sensitivity and repressed c-fos transcription in LM cells but not in AT cells. The results may be a. reason of increased radiation sensitivity of AT cells. PKCI may be involved in an ionizing radiation induced signal transduction pathway responsible for radiation sensitivity and c-fos transcription. The data also provided evidence for novel transcriptional difference between LM and AT cells.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.61-61
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• 2001

The main goal of this study was to produce transgenic piglets by the method of injection of sperm-mediated exogenous DNA. Spermatozoa (1$\times$106 sperm of final concentration) obtained from caudal epididymis were mixed with pBC1-hEPO (20 ng/${mu}ell$) or pcDNA3 LAC Z (20 ng/${mu}ell$), and followed by electroporation (500 V, 25 ㎌). Matured oocytes having the first polar body and dense cytoplasm were selected and centrifuged at 12,000g for 6 min. After sperm injection, the oocytes were activated electrically (1.7 ㎸/cm, 30 $\mu$ sec, single pulse) in 0.3 M mannitol solution. Eggs injected sperm were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air for 192 h. This study were comprised 3 experiments. Experiment 1 compared the developmental efficiencies between the sperm-injected oocytes (Group 1) and further activated electrically (Group 2). Experiment 2 compared the expression of pcDNA3 LAC Z in the embryos produced by Group 1 and Group 2. Finally, experiment 3 carried out transfer of embryos (1-8 cell stage) transfected with pBC1 -hEPO into surrogate recipients synchronized by injection of combination of PG600 with hCG. The rates of cleavage and development into blastocyst stage in Group 2 were significantly higher than those of Group 1 (71.3% and 28.1% vs. 43.3% and 10.3%, respectively, p<0.05). Thirty (24.2%) out of 124 embryos analyzed in Group 2 were positive by X-gal. Similarly, in Group 1, 16.3% (8/49) were positive. After transfer of 789 embryos to 7 recipient gilts, three out of them examined by ultrasound became pregnant. One recipient is in day 50 pregnancy. On day 54 of gestation, two were carried out uterotomy in order to confirm the pregnancy One had 7 and another had 2 fetuses. We conclude that injection of sperm-mediated gene transfer will be used as a valuable tool for the production of transgenic piglets.

• Physical Therapy Rehabilitation Science
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• v.5 no.2
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• pp.89-94
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• 2016

Objective: This study was aimed to compare the thickness and pennation angle of gastrocnemius through ultrasonography during the heel-drop exercise on ankle dorsiflexion angle. Design: Cross-sectional study. Methods: Nineteen normal adults in their 20s had voluntarily participated in this study. All subjects performed the ankle heel-drop exercise with ankle dorsiflexed to $0^{\circ}$, $10^{\circ}$, and $20^{\circ}$: heel-drop exercise with ankle dorsiflexed to $0^{\circ}$ was executed on floor-level, heel-drop exercise with ankle dorsiflexed to $10^{\circ}$ on a wooden-block of 2.3 cm in height, and heel-drop exercise with ankle dorsiflexed to $20^{\circ}$ on a wooden-block of 5.5 cm in height. In each regimen, the subjects completed a session of 100 heel-drop exercises (10 repetitions${\times}$10 sets; with 30 seconds rest following each set; with 24 hours rest following each exercise). Before and immediately after each heel-drop exercise, the thickness and pennation angle of gastrocnemius were measured using an ultrasonography. Results: After the performance of the heel drop exercises with ankle dorsiflexed to $0^{\circ}$, $10^{\circ}$, and $20^{\circ}$, the thickness of the gastrocnemius was significantly higher than pre-exercise (p<0.05), and furthermore heel-drop exercise with ankle dorsiflexed to $10^{\circ}$ was significantly higher than exercise with the ankle dorsiflexed to $0^{\circ}$ (p<0.05). However, as for the pennation angle of the gastrocnemius, there were no significant changes after each heel-drop exercise. Conclusions: This finding suggest that the heel-drop exercise with ankle dorsiflexed to $0^{\circ}$, $10^{\circ}$, and $20^{\circ}$ is effective on the strengthening of the gastrocnemius. Furthermore, the heel-drop exercise with the ankle dorsiflexed to $10^{\circ}$ is more effective than with the ankle dorsiflexed to $0^{\circ}$.

• Microbiology and Biotechnology Letters
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• v.36 no.3
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• pp.221-226
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• 2008

To produce human ferritins in yeast, human H-chain and L-chain ferritins were amplified from previously cloned vectors. Each amplified ferritin gene was inserted into the pYES2.1/V5-His-TOPO yeast expression vector under the control of the GAL1promoter. Western blot analysis of the recombinant yeast cells revealed that H-and L-chain subunits of human ferritin were expressed in Saccharomyces cerevisiae. Atomic absorption spectrometry (AAS) analysis demonstrated that the intracellular content of iron in the ferritin transformant was 1.6 to 1.8-fold higher than that of the control strain. Ferritin transformants could potentially supply iron-fortified nutrients for food and feed.

• BMB Reports
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• v.28 no.3
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• pp.243-248
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• 1995

Cholesteryl ester transfer protein (CETP), a hydrophobic glycoprotein promoting transfer of cholesteryl esters (CE) from high-density lipoproteins (HDL) to lower-density lipoproteins in the plasma, has been recognized a potent atherogenic factor during the development of coronary artery diseases. This study demonstrated a possible utilization of antisense RNA to inhibit expression of the CETP gene using vaccinia virus as an expression system. The CETP cDNA was inserted into a transfer vector (pSC11) in sense and antisense orientations and used to generate recombinant viruses. Recombinants containing sense or antisense orientations of the CETP cDNA were isolated by $TK^-$ selection and X-gal test. The inserted CETP cDNAs in the recombinants were identified by Southern blot analysis and allowed to transcribe in host cells (CV-1). Expressions of the exogenous CETP mRNA, extracted from the CV-1 cells coinfected with viruses containing sense and antisense DNAs, were monitored by Northern blot analysis using the CETP cDNA probe, by Western blot analysis using monoclonal antibody against the C-terminal active region of the CETP and by the CETP assay. Decreased expressions of the exogenous CETP cDNA were clearly evident in the Northern and Western blot analyses as the dose of antisense expression increased. In the CETP assay, the CETP activities decreased compared to the activity obtained from the cell extracts infected with sense construct only.

• 한국신재생에너지학회:학술대회논문집
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• 2011.05a
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• pp.171-171
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• 2011

리그노셀룰로오스 바이오매스를 원료로 하여 발효당을 생산하고자 할 때 최종적인 당수율은 적용하는 전처리 기술의 종류와 전처리 조건에 따라 크게 달라질 수 있다는 사실은 널리 알려져 있다. 또한 전처리물의 효소당화에서 셀룰로오스의 당전환율은 효소의 종류와 사용량에 의존적이므로 부족한 전처리 효과를 일부 보완하기도 한다. 본 연구에서는 바이오매스의 효소당화에 흔히 사용되고 있는 특정효소와 최근 공급되기 시작한 새로운 효소를 시료로 하여 해바라기 줄기의 열수전처리에 이은 효소당화에서 효소의 종류가 최종 당수율, 효소당화 시간 및 전처리효과에 미치는 영향을 측정하였다. 해바라기 줄기 분말을 $180^{\circ}C$에서 30분 동안 열수전처리하였을 때 헤미셀룰로오스의 수율이 최대가 되었으나 이 조건에서는 후속 효소당화에 의한 포도당 수율이 높지 않았다. Celluclast 1.5L 혹은 Celluclast conc BG는 전처리 물의 당화속도가 상대적으로 빠른 편이었고, 이 효소에 의한 당수율은 해바라기 줄기 셀룰로오스 함량의 80% 내외였으며, 바이오매스 1g당 효소첨가량이 3ml까지 증가함에 따라 당수율도 꾸준히 증가하는 경향을 보였다. 반면에 노보자임 스코리아로부터 분양받은 효소로서 NS22074와 NS50010의 혼합물은 Celluclast보다 약 10% 더 높은 당수율을 보여주었으나 당화는 상대적으로 느리게 진행되어 72시간 이상이 소요되었다. Endoxylanase 혹은 hemicellulase 등 다른 효소를 NS22074와 NS50010의 혼합물에 가하여도 당수율의 증대 효과는 미미하거나 거의 없었다. 시험에 사용한 효소제제에는 포도당, 소르비톨 등 여러 가지 당들이 보존제 혹은 안정화제로 함유되어 있는 것으로 나타나서 사용에 주의할 필요가 이었다.