• YAKHAK HOEJI
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• v.48 no.3
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• pp.189-196
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• 2004

Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) playa key role in tumor invasion and metastasis. As an inhibitor of MMP-2, TIMP-2 is known to block both the invasive and metastatic behavior of cancer cells, and decrease tumor growth activity. We performed this study to investigate the effects of TIMP-2 over-expression induced by retroviral mediated gene transfer in vitro and in vivo. The human colon cancer cell line SW480 was transfected with the retroviral vector encoding TIMP-2. The effects of TIMP-2 over-expression were analyzed by invasion assay and gelatinase activity test in colon cancer cells and tumorigencity in nude mice. In evaluation of the transfection efficiency of the retroviral vector encoding TIMP-2 in colon cancer cells, we confirmed up-regulation of TIMP-2 expression dependent on the time of cell culture. In addition, inhibition of MMP-2 expression in SW480/TIMP-2 was shown by gelatin zymography. In the in vitro invasion assay SW480/TIMP-2 inhibited the invasiveness on matrigel coated with collagen. To determine whether TIMP-2 can modulate in vivo tumorigenicity and metastasis, SW480/TIMP-2 cells were injected subcutaneously in nude mice. The tumor mass formation of SW480/TIMP-2 cells in nude mice was markedly decreased compared to nontransfected cancer cells. These results showed that colon cancer cells transfected with the retroviral vector encoding TIMP-2 inhibits the invasiveness in vitro and tumorigenicity in vivo.

• Journal of the Korean Recycled Construction Resources Institute
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• v.10 no.3
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• pp.300-306
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• 2022

The distribution properties of airborne chlorides at the coasts of Jeju island were identified in this study. For this, Jeju island was divided into four regions-east, west, south, and north, and the airborne chlorides were investigated in those regions. And the regional and seasonal distribution properties of airborne chlorides were analyzed. Jeju island, which has a subtropical climate and is surrounded by ocean, has a great regional variation in the distribution properties of airborne chlorides, with a difference of more than 6 times between the eastern region, the highest region, and the southern region, the lowest region. In terms of seasonal properties, the airborne chlorides in eastern and western region were high in summer and fall, and in summer mainly due to typhoon. The airborne chlorides in northern region were high in winter due to the northwest wind, and the that of southern region were high in summer. Meanwhile, the airborne chlorides of Jeju island tends to be high compared to the eastern, western, and southern coasts of South Korea.

• Current Research on Agriculture and Life Sciences
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• v.8
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• pp.45-50
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• 1990

This study was carried out ot investigate the effects of fetal calf serum (FCS) and gonadotropins supplemented to the medium on maturation and fertilization in vitro of porcine follcular oocytes. Ovaries were obtained from gilts at local slaughter-house. Oocyte-cumulus complexes were recovered by puncturing the ovarian follicles(3~5 mm in diameter). The complexes from individual ovaries were pooled in a $0.4m{\ell}$ droplet of medium covered with paraffin oil, then washed twice in fresh droplet and cultured for 36hrs in culture media according to experimental conditions. Boar epididymal spermatozoa were capacitated by preincubation for 4hrs in m-KRB medium and the preincubated spermatozoa were insemenated in the fertilization medium containing the cultured oocytes. The results obtained in this study are summarized as follows: 1. The maturation rates of oocytes cultured in m-KRB and m-KRB supplemented to 10% FCS were 82 and 37%, respectively. When PMSG, hCG. and PMSGt hcG($10Iu/m{\ell}$) were added to the media supplemented to 10% FCS, the maturation rates were 66, 58 and 68%, respectively. 2. Expansion of cumulus cells was not occured in m-KRB and m-KRB supplemented to 10% FCS. However, when PMSG, hCG and PMSG+hCG($10Iu/m{\ell}$) were added to m-KRB supplemented to 10% FCS, the expansion rates of cumulus cell layers were 92, 13 and 91%, respectively. 3. When oocytes were mltured in m-KRB, the rates of penetration and formation of male pronucle: were 93 and 7%, respectively. By adding FCS and gonadotropin to m-KRB, the penetration and formation of male pronuclei were 100 80%, respectively.

• Korean journal of applied entomology
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• v.59 no.1
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• pp.25-35
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• 2020

The striped fruit fly, Zeugodacus scutellata, is endemic in Korea, but it has been regarded as one of the serious quarantine pests throughout the world. Sterile insect release technique (SIT) has been used to eradicate quarantine fruit flies. This study developed a technique to generate sterile males and applied SIT to control Z. scutellata. First of all, the reproductive systems of Z. scutellata adults were examined with fluorescent microscope. Polytrophic ovaries comprises of around 100 follicles with developing oocytes. Each follicle contains an oocyte with several nurse cells and are surrounded with follicular epithelium. Oocyte development began at 10 days after adult emergence (DAE) and formed chorionated oocytes after 20 DAE. On the other hand, male testes were well developed just after adult emergence. The vas deferens was filled with motile sperms. To generate sterile males, different doses (0~1,000 Gy) doses of electron beam were irradiated to 3~5 days old pupae of Z. scutellata. When male pupae were irradiated with electron beam at 200 Gy, they developed and mated with females without any significant difference compared to untreated males. Although the untreated females mated with the 200 Gy-irradiated males laid eggs, no eggs did not hatch. The 200 Gy-irradiated males were then applied to untreated male and female flies in a density ratio of 1:9 (untreated males : treated males). The laid eggs suffered significant infertility. These results suggest that electron beam-irradiated pupae at 200 Gy resulted in male sterility and the resulting males would be applied to SIT.

• Journal of Aquaculture
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• v.20 no.4
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• pp.219-225
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• 2007

This study investigated the correlationship between artificial maturation season and reproduction coefficient of cultured eel Anguilla japonica from May (spring) to next January (winter). The brood stock, female eels ($400{\sim}600\;g$) were artificially matured by weekly intramuscular injections of salmon pituitary extracts (SPE, 20 mg/fish) to induce a completion of vitellogenesis. After completion of vitellogenesis, final oocyte maturation and ovulation was induced by injection of $17{\alpha}$, $20{\beta}-dihydroxyprogesterone$ (DHP) at about $2\;{\mu}g/g$ body weight. Most fish ovulated $15{\sim}18\;h$ following the DHP injection. The ovulated fish were induced to natural spawning or artificial fertilization by the dry method. Males ($200{\sim}350\;g$) were received weekly intramuscular injections of human chorionic gonadotropin (HCG) at a dosage of 1 IU/g body weight to induce testicular maturation and spermiation. Seasonal reproduction coefficient which includes the rate of ovulation, buoyancy, fertilization and hatching of eggs in the artificially matured eel during spring to summer ($May{\sim}July$) were significantly higher than the other season, while there were no significant difference among spring and summer (P<0.05). Furthermore, the number of eggs spawned and larvae hatched in the artificially maturated eel during spring to summer ($May{\sim}July$) were significantly higher than the other season, while there were no significant difference in spring and summer (P<0.05). These results indicate that artificial maturation by hormone treatment of A. japonica was successful only during spring to summer, which is the maturation period in the wild stock in nature. Consequently, it is possible to determine the period of artificially induced sexual maturity by the reproduction coefficient which includes the rate of ovulation, buoyancy, fertilization and hatching of eggs in the cultured eel A. japonica.

• The Journal of the Petrological Society of Korea
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• v.19 no.2
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• pp.109-121
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• 2010

Mt. Mudeung is located in Gwangju city, Damyang-Gun, Hwasun-Gun and its round form give us the mood of soft and rich. Its location is $126^{\circ}06'-127^{\circ}01'E$ and $35^{\circ}06'-35^{\circ}10'N$ and its highest peak is Cheonwang-bong with the height of 1,187 m. The Gwangju city is located in the West of Mt. Mudeng and the mountain range with a small basin in its East. The pavilion such as the Soswaewon, Songganjeong, Sigyongjeong are distributed along the stream in the north of Mt. Mudeung. The mountain is formed from the volcanic activity, Gwangju cauldron during the Cretaceous. The top part of Mt. Mudeung is composed of dark gray quartz-andesite and its K-Ar whole rock age is $48.1{\pm}1.7Ma$. The composition of the north area, where the Wonhyosa temple is located, is micrographic granite, whereas the composition of south area is rhyolite mainly. The main ridge of Mt. Mudeung runs from North, starting from the Bukbong, to south, passing Cheonwangbong, Jangbuljae and ending Anyangsan. Geologic feature of the mountain includes volcanic landform, mountaineous landform, and stream landform. The Seosukdae, Ipseokdae, Gyubongam, which are main ridges and formed from volcanic activity, are composed of mainly columnar joint. Saeinbong and Majipbong in the south-west are composed of mainly cliff and dome. The typical erosion landform of the mountain has three different types of the weathering-cave, each of which reflect the property of the original rock. Four different area of wide block stream, they makes the geological feature of spring-water, though its scale is small compared to that of water fall.

• Journal of Life Science
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• v.23 no.9
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• pp.1133-1139
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• 2013

During the collection of boar semen, bacterial contamination usually occurs. The contamination has deleterious effects both on semen quality and on sow fertility. The majority of contaminants are gram-negative bacteria, especially Serratia marcescens. In this study, we developed a PCR assay for the identification of S. marcescens targeting the luxS gene (GenBank no. EF164926). S. marcescens yielded a specific 306 bp PCR product. However, no amplification was observed in the other strains tested. The detection limit of PCR was $50pg/{\mu}l$ of template DNA of S. marcescens. The antimicrobial susceptibility patterns of S. marcescens isolated from boar semen were tested using the disk diffusion method. Gentamicin, ceftiofur, florfenicol, and neomycin showed high sensitivity in this test. The minimum inhibitory concentration (MIC) was also determined by the broth microdilution method. The $MIC_{90}$ values of ceftiofur, enrofloxacin, gentamicin, and neomycin were 8, 8, 8, and $16{\mu}g/ml$, respectively. These results indicate that PCR amplification of the luxS gene is a reliable and effective method for the identification of S. marcescens and that ceftiofur, enrofloxacin, gentamicin, and neomycin are effective semen extenders for controlling S. marcescens.

• Korean Journal of Environmental Biology
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• v.20 no.3
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• pp.245-255
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• 2002

In order to obtain the basic information for seedling production of venus clam, Protothaca jedoensis, the annual reproductive cycle was investigated mainly by histological observation from September 1998 to August 1999. P. jedoensis was dioecious. The gonads are located between the digestive diverticula and muscle tissue of the foot. The ovary was composed of a number of ovarian sacs, and the testis was composed of several testicular tubules. The condition index was reached its maximum (20.0) in February, and then decreased to 11.5 in June. In August, the value was the lowest (9.0) and then increased slowly. Minimum size for the sexual maturation of individuals were 38.4 mm in shell length. One the other hand, the size of mature oocytes was ranged to $50-60\mu{m}$ in diameter and testis-ova was observed in testis of the mature stage. The reproductive cycle of P. jedoensis could be classified into five successive stages: early active (December to February), late active (January to April), ripe (March to July), partially spawned (June to August) and spent/inactive (July to January) stages.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.38-38
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• 2001

Spermatogonial stem cells은 sperrnatogenesis에서 중요한 역할을 하며, 곡세정관의 기저막에 위치하고 있는 것으로 알려져 있다. 그러나, 그 동안 이 세포에 특이하게 발현되는 marker가 거의 알려져 있지 않아 spermatogonial stem cell의 연구에 많은 어려움을 가져왔다. 최근 일반적인 stem cell이 갖는 특성 중, 기저막과 상호작용을 하는 surface protein으로 integrin이 존재한다는 사실을 이용하여, anti-$\alpha$$_{6}$/ 또는 anti-$\beta$$_1$ integrin항체로 germ cell을 선발하여 정소에 이식한 결과, 높은 효율로 이식세포유래의 정자발생이 가능하다는 결과가 보고되었다 (Shinohara et al., 1999). 한편, 항암제의 일종인 busulfan을 마우스에 투여(40mg/kg)한 후 4-5주가 경과하면 세정관의 기저막에 위치하는 spermatogonia를 제외하고 대부분의 생식세포는 소멸한다 본 실험의 목적은 이러한 사실들을 이용하여 spermatogonial stem cell의 특성을 밝히고, 이 생식세포를 보다 간편하고 손쉽게 선발할 수 있는 시스템을 확립하는데 있다. Busulfan처리 후 5주가 경과된 마우스와 정상적인 13주령의 마우스 testis로부터 세포를 분리한 후 FITC-conjugated anti-IgG를 이용한 면역형광법으로 측정.분석한 결과, 형광표식된 세포비율이 대조군과 비교하여 busulfan을 처리한 경우에서 유의적인 증가를 보였다.(17$\pm$3.8%. 0.7$\pm$0.3% busulfan vs control). 또한, IgG와 결합한 단백질이 존재하는 이들 세포들은 곡세정관의 기저막을 따라 위치하며, 단백질과 복합체를 형성한 IgG는 anti-Ig $G_{2a}$와 반응하지 않는다는 사실을 관찰했다. 이러한 IgG 복합체를 형성한 세포들의 특성을 이용하여, IgG와 반응을 하지 않는 것으로 확인된 이차 항체인 an1i-Ig $G_{2}$와 일차 항체인 anti-$\alpha$$_{6}$ 또는 anti-$\beta$$_1$ integrin항체를 이용하여 측정.분석하였다. Busulfan을 처리한 마우스 정소에서 분리한 세포를 다시 laminin으로 코팅된 dish에서 선발.회수해서, anti-lgG, anti-$\alpha$$_{6}$ 또는 anti-$\beta$$_1$ integrin항체로 각각 표식된 세포비율을 비교하였다. Laminin으로부터 선발.회수한 세포에서는 IgG복합체가 $\alpha$$_{6}$ 또 는 $\beta$$_1$integrin과 거의 같은 수준에서 높은 비율로 표식되었다. 결론적으로, busulfan에 의해 유도된 IgG와 결합가능한 단백질은 $\alpha$$_{6}$나 $\beta$$_1$ integrin과 마찬가지로 immunoglobulin G를 이용하여 spermatogonial stem cell의 선발을 가능하게 했다. 따라서, busulfan처리시 IgG는 미분화된 정조세포의 선발을 위한 하나의 marker로서 사용가능함을 시사한다.다.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.82-82
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• 2002

Catalase(CAT)는 주요한 활성산소(ROS)의 일종인 과산화수소($H_2O$$_2$)가 Hydroxyl 유리기 생성에 관여하지 못하도록 $H_2O$$_2$를 $H_2O$ 및 $O_2$로 대사시켜 주는 효소계 항산화제의 한 종류이다. 따라서 본 실험에서는 CAT가 5% $O_2$및 20% $O_2$조건하에서 1-세포기 돼지체외수정란의 배발달에 미치는 영향을 조사하였다. 돼지난포란을 10% PFF, 0.1mg/ml cysteine, 10IU/m1 PMSG, 10IU/m1 hCG 및 10ng/m1 EGF가 첨가된 NCSU23 배양액에서 22시간 동안 배양을 실시하고, 성선자극호르몬이 배제된 배양액에서 추가로 22시간을 배양하여 체외성숙을 유도하였다. 체외성숙이 유기된 난자는 난구세포를 제거하고, 2.5mM caffeine과 0.1% BSA가 첨가된 mTBM 배양액에 정자를 1.25 $\times$ $10^{5}$cells/ml의 농도로 5-6시 동안 공동배양을 실시하여 체외수정을 유도하였다. 체외수정후 1-세포기의 수정란을 0.4mg/ml BSA가 첨가된 NCSU23 배양액에 CAT를 각각 0, 100, 500 및1,000uni1 첨가하여 30 embryos/ 50u1 소적으로하여 38.8$^{\circ}C$, 5% $CO_2$및 5%$CO_2$, 5% $O_2$90% $N_2$조건하의 배양기에서 각각 7일간 배양을 실시하였다. 조사된 결과는 SAS/STAT 6.03 Package를 이용하여 통계분석을 실시하였다. 체외배양 48시간에 난할률을 조사하였을 때, 대조구와 처리구간 차이가 인정되지 않았으나, 배양 7일째 배반포형성률에 있어서는 각각 22.7$\pm$2.7, 22.1$\pm$3.9, 18.7$\pm$4.9, 및 15.1$\pm$2.5%로서 처리구가 대조구보다 유의적으로 낮은 결과를 나타냈다. 그리고 산소농도에 따른 배반포형성률은 5% $O_2$ 조건(22.1$\pm$2.4%)이 20% $O_2$조건(17.2$\pm$2%)보다 유의적(P<0.05)으로 높은 것으로 나타났다. 총세포수에 있어서는 각각 44.4$\pm$4.0, 43.3$\pm$36, 25.4$\pm$2.4 및 13.4$\pm$1.5로서 처리구가 대조구보다 세포수가 적은 것으로 나타났으며, 산소농도에 따른 차이는 인정되지 않았다. 따라서, 체외생산된 돼지초기수정란을 배양할 때, CAT의 첨가는 효과가 없는 것으로 나타났다.다.