• 생명과학회지
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• 제10권2호
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• pp.150-156
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• 2000

This experiment was performed to evaluate the effect of TSH (thyroid-stimulating) on the ERp29 (endoplasmic reticulum resident 29 kDa protein) gene expression in the rat thyrocytes of FRTL-5 cells. Although ERp29 mRNA was constantly expressed, its expression began to increase remarkably from 10-9 M TSH. and its maximum expression was at 5×10-9 M TSH (about 3.5 fold). On the other hand, the effect of TSH on the abundance of ERp29 mRNA started within 6 h, and peaked at 8 h (about 2.5 fold). Actinomycin D (transcription inhibitor) strongly blocked this effect while cycloheximide (translation inhibitor) did not. The half-life of ERp29 mRNA was about 4.5 h in the presence or absence of TSH that was not affected by the stability of ERp29 mRNA. The effect of TSH on the ERp29 gene expression was specific, while other growth factors (transfferin, insulin, and hydrocortisone) did not alter its expression. Our data indicate for the first time that the expression of ERp29 is regulated transcriptionally by TSH in the thyrocytes.

• 한국미생물·생명공학회지
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• 제18권1호
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• pp.26-30
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• 1990

포도에서 분리하여 동정한 Candida dattila K109와 K112 균주는 Kluyveromyces, Hansenula, Debaryomyces, Torulopsis, Brettanomyces속 효모에 대해 killer 활성을 가졌으며, 이들 균주의 최적 pH는 3.9-4.0이고, 최적 온도는 22-26$^{\circ}C$였다. Candida dattila K109와 K112 균주의 toxin은 단백질 분해효소인 pronase E와 pepsin에 의한 killer 활성이 없어졌으며, 2$0^{\circ}C$에서는 비교적 안정하였으나 $25^{\circ}C$ 이상에서는 killer 활성이 급격히 감소하였으며, pH 2.0-4.0 범위에서 비교적 안정하였고 그 이상의 pH에서는 급격히 활성이 저하되었다. 이들 균주의 killer toxin은 gel filtration에 의하여 단백질과 당을 확인하였다. Candida dattila K109와 K112 균주는 0.0105-0.3ppm cycloheximide 처리에 의하여 처리농도가 놓을 수록 killer 활성이 소멸되는 비율이 증가하였으며, 30-37$^{\circ}C$의 가온처리에 의하여 killer활성이 소멸되지 않는 등 기존의 killer 효모의 특성과 다소 다른 특성을 나타내었다.

• The Plant Pathology Journal
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• 제21권4호
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• pp.301-309
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• 2005

Trichoderma species/isolates exhibited varied degree of agglutination on sclerotial (Sc) and hyphal (Hy) surface of Macrophomina phaseolina. The agglutination efficiencies on Sc and Hy ranged from $11\;to\;57\%$. Isolates of T. harzianum (Th) and T. viride (Tv) showed greater agglutination on Sc ($23-57\%$) and Hy ($16-47\%$). Different enzymes (trypsin, pepsin, proteinase k, a-chymotrypsin, lyticase and glucosidase) and inhibitors (tunicamycin, cycloheximide, brefeldin A, sodium azide, dithiothreitol and SDS) reduced the agglutination potential of conidia of Th-23/98 and Tv-25/98; however, the extent of response varied greatly in different treatments. Different fractions of Th-23/98 and Tv-25/98 exhibited haemagglutinating reaction with human blood group A, B, AB and O. Haemagglutinating activity was inhibited by different sugars and glycoproteins tested. Crude haemagglutinating protein from outer cell wall protein fraction of Th-23/98 and Tv-25/98 were eluted on Sephadex G-100 column. Initially Th-23/98 and Tv-25/98 exhibited two peaks showing no agglutination activity; however, lectin activity was detected in the third peak. Similar to crude lectin, the purified lectin also exhibited haemagglutinating activity with different erythrocyte source. SDS-PAGE analysis of partially purified lectin revealed single band with an estimated molecular mass of 55 and 52 kDa in Th-23/98 and Tv-25/98, respectively. Trypsin, chymotrypsin and b-1,3-glucanase totally inhibited lectin activity. Similarly, various pH also affected the haemagglutinating activity of Th-23/98 and Tv-25/98. From the present observations, it can be concluded that the recognition/attachment of mycoparasite (T. harzianum and T. viride) to the host surface (M. phaseolina) may be most likely due to lectin-carbohydrate interaction.

• 한국동물학회지
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• 제30권2호
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• pp.193-209
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• 1987

Patterns of amylase secretion in mouse pancreatic fragments were studied over a period of time after the tissue was stimulated by acetyicholine and MNNG. MNNG is known to activate guanylate cyclase and thus increase the cGMP concentration in the pancreatic acinar cell. These amylase secretion patterns were studied to investigate the role of cGMP in reaction cascade during secretion response of the tissues stimulated by acetyicholine. Cellular response of amylase secretion in the pancreas by acetyicholine was divided into two phases. During the first phase, zymogen granules which had existed in the cells were secreted by the action of $Ca^2$+ and calmodulin immediately after secretagogue administration, this being known as the initial response. When the tissue was stimulated by acetylcholine in a $Ca^2$+-deficient medium or one containing trifluoperazine as a calmodulin antagonist, this initial response was reduced. In the second phase, newly formed zymogen granules were secreted as sustained response after protein synthesis was triggered by secretagogue. This response was provoked by an activation of protein kinase C. When either cycloheximide as a protein synthesis inhibitor or dibucaine as a protein kinase C inhibitor were added to the incubation medium, this sustained response was remarkablely depressed in the pancreatic fragments stimulated with acetylcholine. In the pancreatic acinar cell, phosphatidylinositol turnover plays an important role in the secretion response and hexachlorocyclohexane inhibits this phosphatidylinositol turnover. The pancreatic tissue treated with the hexachlorocyclohexane exhibited inhibition on both initial and sustained responses of amylase secretion by acetylcholine. MNNG also accelerated amylase secretion from the tissue gradually along incubation time. The 22 minutes fraction of the pancratic secretion after administration of both acetylcholine and MNNG showed higher amylase activity than the neighboring fractions. Guanylate cyclase potentiated the sustained response. Even if it is experimented with an indirect method, guanylate cyclase was found responsible for activation of the sustained response of a step prior to the action of protein kinase C. As conclusion, it was considered that amylase secretion in mouse pancreatic fragments stimulated by acetylcholine is a three phasic response.

• Animal cells and systems
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• 제14권3호
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• pp.147-154
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• 2010

It is well-established that phosphoinositide 3-kinase (PI3-kinase) regulates myogenesis by inducing transcription of myogenin, a key muscle regulatory factor, at the initiation of myoblast differentiation. In this study, we investigated the role of PI3-kinase in cells that have committed to differentiation. PI3-kinase activity increases during myogenesis, and this increase is sustained during the myogenic process; however, its function after the induction of differentiation has not been investigated. We show that LY294002, a PI3-kinase inhibitor, blocked myoblast fusion even after myogenin expression initially increased. In contrast to the inhibitory effects of LY294002 on myogenin mRNA levels during the initiation of differentiation, LY294002 blocked the accumulation of myogenin protein without affecting its mRNA level after differentiation was induced. Treatment with cycloheximide, a translation inhibitor, or actinomycin D, a transcription inhibitor, indicated that the stability of myogenin protein is lower than that of its mRNA. LY294002 inhibited the activities of several important translation factors, including eukaryotic elongation factor-2(eEF2), by altering their phosphorylation status. In addition, LY294002 blocked the incorporation of [$^{35}S$]methionine into newly synthesized proteins. Since myogenin has a relatively short half-life, LY294002-mediated inhibition of post-transcriptional processes resulted in a rapid depletion of myogenin protein. In summary, these results suggest that PI3-kinase plays an important role in regulating the expression of myogenin through post-transcriptional mechanisms after differentiation has been induced.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.28-28
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• 2003

Haploid parthenotes have been shown to be developmentally delayed compared with diploid parthenogenetic embryos in the mouse and pig. These developmental defects have been hypothesized to rusult from insufficient parthenogenetic activation, suboptimal in vitro culture conditions, or genemic imprinting. In the present study we compared the incidence of apoptosis and apoptosis related gene expression in pig haploid, diploid parthenotes and fertilized embryos. In vitro matured porcine oocytes were activated by electrical stimulation. Haploid activated oocytes with two polar bodies under stereomicroscopy were defined haploid parthenotes, oocytes with one polar body were defined as diploid parthenotes after 3h cycloheximide teatment. The morphological analysis of apoptosis in embryos was carried out using propidium iodide staining and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling. The expression of Bcl-xL, Bak and P53 in haploid, diploid and in vivo fertilized blastocysts was determined using RT-PCR. Lower number of the haploid pig parthenotes developed to the morulae and blastocysts compared to the diploid parthnotes. Number of cells significantly lower in the haploid-derived blastocysts than diploid-derived it. Developmentally retarded haploid parthenotes exibited apoptosis at a significantly higher frequency than did diploid parthenotes and fertilized embryos. Level of Bcl-xL expression, diploid parthenotes similar to in vivo-derived it was higher than haploid parthenotes. However, Bak and P53 mRNA expression were not different among haploid, diploid, and fertilized embryos. This result suggested that parthenogenetic activation and parthenogenesis themselves do not cause apoptosis, but haploid increases the incidence of apoptosis in preimplantation embryos. Apoptosis may be due to decrease expression of Bcl-xL in haploid parthenotes developing in vitro.

• Journal of Ginseng Research
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• 제48권1호
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• pp.40-51
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• 2024

Background: Ginsenoside 20(S)-Rg3 shows promising tumor-suppressive effects in ovarian cancer via inhibiting NF-kB signaling. This study aimed to explore the downstream tumor suppressive mechanisms of ginsenoside Rg3 via this signaling pathway. Materials and methods: A systematical screening was applied to examine the expression profile of 41 kinesin family member genes in ovarian cancer. The regulatory effect of ginsenoside Rg3 on KIF20A expression was studied. In addition, we explored interacting proteins of KIF20A and their molecular regulations in ovarian cancer. RNA-seq data from The Cancer Genome Atlas (TCGA) was used for bioinformatic analysis. Epithelial ovarian cancer cell lines SKOV3 and A2780 were used as in vitro and in vivo cell models. Commercial human ovarian cancer tissue arrays were used for immunohistochemistry staining. Results: KIF20A is a biomarker of poor prognosis among the kinesin genes. It promotes ovarian cancer cell growth in vitro and in vivo. Ginsenoside Rg3 can suppress the transcription of KIF20A. GST pull-down and co-immunoprecipitation (IP) assays confirmed that KIF20A physically interacts with BTRC (β-TrCP1), a substrate recognition subunit for SCF^β-TrCP E3 ubiquitin ligase. In vitro ubiquitination and cycloheximide (CHX) chase assays showed that via interacting with BTRC, KIF20A reduces BTRC-mediated CDC25A poly-ubiquitination and enhances its stability. Ginsenoside Rg3 treatment partly abrogates KIF20A overexpression-induced CDC25A upregulation. Conclusion: This study revealed a novel anti-tumor mechanism of ginsenoside Rg3. It can inhibit KIF20A transcription and promote CDC25A proteasomal degradation in epithelial ovarian cancer.

• 대한핵의학회지
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• 제34권3호
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• pp.252-259
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• 2000

목적: 핵의학과에서 흔하게 이용되는 Tc-99m MDP 또는 Tc-99m DTPA를 주사한 후 방사선 적응반응이 일어나는지를 확인하고, 어느 경우에 방사선 적응반응이 더 현저하게 유도되는지 알아보기 위하여 이 연구를 시행하였다. 대상 및 방법: Tc-99m MDP 또는 Tc-99m DTPA 신티그라피를 시행한 45명의 환자(남자 25명, 여자 20명, 평균 연령 $44{\pm}18$세)를 대상으로 Tc-99m MDP 뜨는 Tc-99m DTPA 주사 전과 주사 후 4시간에 각각 5 ml씩 채혈하여 배양하고 배양 46시간 후에 Cs-137조사기 (central dose rate=654 Gy/h, Gammacell 3000 Elan, Nordion, Canada)를 이용하여 2 Gy의 감마선을 조사하였다. 대조군 20명(남자 12명, 여자 8명, 평균연령 $43{\pm}7$세)의 혈액을 채혈하여 Tc-99m MDP 또는 Tc-99m DTPA 주사 전에 채혈한 군과 같은 방법으로 조사하고 배양하였다. Colcemid 처리 2시간 후에 수확하여 불안정 염색체인 반지형과 이중 중심체형 염색체의 수자를 계수 하여 불안정 염색체 출현빈도(Ydr)를 구하여 비교하였다. 고선량의 방사선 단독조사에 의한 Ydr값과 방사성의약품 투여 후 고선량의 방사선 조사에 의해 유도된 Ydr값의 차이값을 적응반응지수로 정의하고, Tc-99m MDP 신티그라피와 Tc-99m DTPA 신티그라피의 적응반응지수를 비교하였다. 결과. 2 Gy 단독조사에 의한 Ydr값에 비해 Tc-99m MDP 또는 Tc-99m DTPA 주사한 후에 2 Gy 조사한 군에서 Ydr값이 유의하게 감소하였다($0.45{\pm}0.20\;vs.\;0.24{\pm}0.10$, p=0.001). Tc-99m MDP 또는 Tc-99m DTPA에 의해 유도된 적응반응은 단백합성억제제인 cycloheximide에 의해 억제되었다($0.24{\pm}0.10\;vs.\;0.43{\pm}0.16$, p=0.001). Tc-99m MDP 신티그라피에 의한 적응반응 지수는 Tc-99m DTPA 신티그라피에 의한 적응반응지수에 비해 유의하게 높았다($0.10{\pm}0.06\;vs.\;0.24{\pm}0.05$, p=0.0001). 결론: Tc-99m MDP 신티그라피 또는 Tc-99m DTPA 신티그라피에 의한 저선량의 방사선 조사에 의하여 말초혈액 림프구에서 방사선 적응반응이 유도되었고, 적응반응의 정도는 Tc-99m MDP 골 신티그라피를 시행하는 환자에서 더욱 현저함을 알 수 있었다.

• 대한핵의학회지
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• 제35권2호
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• pp.83-88
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• 2001

목적: 본 연구에서는 분화갑상선암 환자에서 치료용량의 방사성옥소(I-131)를 섭취한 후 말초혈액 림프구에서 방사선적응반응이 유도되는지 알아보고자 하였다. 대상과 방법: 분화갑상선암에 대한 I-131 치료를 시행한 21명의 환자(남자 7명, 여자 14명, 평균연령 $55{\pm}12$세)를 대상으로 I-131 투여전과 5,550 GBq (150 mCi) 투여 후 72 시간에 각각 5 ml씩 채혈하여 배양하고, 배양 70 시간 후에 Cs-137 세포조사기를 사용하여 1 Gy의 감마선을 조사하였다. I-131 치료 후 채혈한 혈액 중 일부는 배양 68 시간째에 단백합성억제제인 cycloheximide (CHM) $10{\mu}g/ml$을 첨가하고 2시간 후에 2차례 세척한 다음 1 Gy의 감마선을 조사하였다. 대조군으로는 염색체 이상을 동반하는 유전병 병력이 없는 건강한 사람 10명(남자 6명, 여자 4명, 평균연령 $43{\pm}7$세)에서 분리한 림프구를 이용하였다. 환자군 중 I-131 치료 전에 채혈한 군과 I-131 치료 후 채혈한 군, 그리고 대조군을 각각 1 Gy를 조사하지 않은 군과 조사한 군으로 나누어 불안정 염색체를 계수한 다음 불안정염색체의 출현빈도를 나타내는 Ydr값을 구하여 서로 비교하였다. 결과: 대조군과 I-131 치료전 환자군에서 Ydr값은 $0.08{\pm}0.01$과 $0.09{\pm}0.01$로 유의한 차이가 없었으나, I-131 치료 후에는 환자군의 Ydr값이 $0.13{\pm}0.02$로 유의하게 증가하였다. 이 혈액에 1 Gy의 감마선을 조사하였더니 Ydr값이 대조군과 환자군의 림프구에서 1 Gy의 감마선을 조사하였더니 Ydr값이 대조군에서는 $0.24{\pm}0.01$이고, I-131 치료전 환자군에서는 $0.21{\pm}0.02$, 치료후 환자군에서는 $0.17{\pm}0.03$로 대조군에 비해 치료전 환자군에서 유의하게 낮았고, 치료후 환자군은 대조군과 치료전 환자군에 비해 더욱 낮았다. 증가치는 대조군 $0.16{\pm}0.01$, 치료전 환자군 $0.12{\pm}0.03$, 치료후 환자군 $0.03{\pm}0.02$로 치료후 환자군에서 유의하게 낮았다. 그러나, I-131 치료후 환자군에 CHM을 처치하고 1 Gy의 외부감마선을 조사한 경우에는 Ydr값이 $0.20{\pm}0.05$으로 CHM을 처치하지 않은 군에 비해 유의하게 높은 값을 보였다. 결론: 고선량의 I-131을 투여한 환자의 림프구에서 방사선적응반응이 유도됨을 알 수 있었고, 이 반응에는 단백질이 관여함을 알 수 있었다.

• 한국수정란이식학회지
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• 제16권2호
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• pp.107-115
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• 2001

본 연구는 최근 형질전환동물의 생산 및 복제동물 생산에 이용되고 있는 핵이식 기법을 시행할 때 재조합된 핵이식란의 활성화를 위해 널리 적용되고 있는 6-dimethylaminopurine (DMAP)의 활성화 효율과 근래에 제기되고 있는 단위발생란의 비정상적인 염색체 및 핵형에 대해 알아보고 적합한 활성화 유도물질을 찾고자 시행되어졌다. 도축장 유래의 난소에서 채란한 난자를 10% 거세한 수소혈청이 포함된 TCM-199배양에서 22시간동안 체외 성숙을 시킨 후 제 2감수분열 중기의 난자만을 선별해서 5$\mu$M ionomycin에서 5분간 처리하고 1.9 mM 6-dimethylaminopurine (DMAP)와 10$\mu\textrm{g}$/mL cycloheximide (CHX)에서 각 3시간동안 처리하여 활성화를 유도하였다. 활성화가 유도된 난자를 18시간 동안 체외 배양시 전핵 형성, 제 2 세포기까지 분할속도, 배 반포까지의 발달을 및 활성화 및 체외수정 후 108시간에 평균 세포수와 염색체를 분석하여 활성화 물질의 효율뿐만 아니라 문제점을 알아보고자 하였다. 1. 활성화 자극에 따른 난자의 전핵 형성은 ionomycin 처리 후 DMAP을 처리한 난자에서는 1PN 형성율이 9.1%로 ionomycin를 단독 처리하거나 ionomycin 처리 후 CHX를 처리한 난자에서의 1PN 형성율인 77.8와 79.0%보다 유의적 (P<0.05)으로 낮게 나타났으나, 3PN 형성율은 45.5%로 유의적 (P<0.05)으로 높게 나타났다. 따라서 ionomycin 처리 후 DMAP으로 활성화를 유도한 난자는 비정상적인 핵형을 가지지만 CHX로 활성화를 유도하였을 때는 정상적인 전핵 형성이 이루어지는 것으로 보인다. 2. 활성화 자극을 가한 난자의 체외 발달율은 ionomycin을 처리하고 DMAP으로 활성화 자극을 가하였을 때 분할율이 85.5%로 체외 수정한 대조군의 72.5%와 유사하였다. 그러나 ionomycin을 단독 처리하거나 ionomycin 처리 후 CHX로 활성화 자극을 가한 실험군의 분할율인 30.3와 57.9%에 비해 유의적 (P<0.05)으로 높게 나타났다. DMAP 처리군의 분할율은 대조군과 유사하였지만 배반포까지의 발달율은 12.3%로 대조군의 27.8%와는 유의적인 차이는 없으나 발달율이 낮은 경향으로 나타났다. 3. Ionomycin으로 처리 후 DMAP로 활성화 자극을 가한 실험군에서 난자의 발달속도는 활성화 자극 후 18시간 경과하였을 때 28%의 배분열율을 보여 분열속도가 가장 빨랐으며 활성화 자극 후 24~48시간동안 체외 배양을 하였을 때에도 ionomycin 단독 처리하거나 ionomycin 처리 후 CHX로 활성화 자극을 준 실험군에 비해 DMAP으로 활성화 자극을 가한 실험군이 유의적 (P<0.05)으로 빠른 발달속도를 보였다.