• Journal of Oral Medicine and Pain
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• 제34권3호
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• pp.261-276
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• 2009

Sreptomyces라는 세균에서 추출한 lactacystin은 선택적인 proteasome 억제제로서 많은 연구에서 사용되어져 왔다. Proteasome 억제제는 최근의 많은 연구를 통해서 암세포증식의 억제에 대한 효과가 증명되었으며, 특히 다른 항암제와 병용처리 시, 상호작용에 의한 상승효과가 있다고 알려져 있다. 현재 proteasome 억제제는 새로운 강력한 항암제로서 분류되어 있다. 본 연구는 사람혀편평세포암종세포(SCC25 cells)에서 lactacystin의 세포독성과 성장억제 효과, 그리고 세포자멸사의 유도에 대한 분자생물학적 기전을 밝히기 위해 실험을 시행하였다. SCC25 세포, 사람정상각화세포 (HaCaT cells) 그리고 사람치은섬유모세포(HGF-1 cells)의 생존율 측정은 MTT법을 시행하였고, SCC25 세포의 성장억제를 확인하기 위해서는 clonogenic assay를 사용하였다. lactcystin이 SCC25 세포에서 세포자멸사가 유도되는지를 확인하기 위해서 hoechst 염색법, hemacolor 염색법 그리고 TUNEL법을 시행하였다. 그리고 SCC25 세포에 lactacystin을 적용한 후, Western blot 분석, 세포면역화학염색, 공초점레이저주사현미경 검경, FACScan flow cytometry, 사립체막 전위변화, proteasome 활성도 측정 등을 시행하였다. Lactacystin으로 처리된 SCC25 세포는 시간 및 용량 의존적인 세포생존율의 감소, 용량의존적인 세포성장억제 그리고 세포자멸사에 의한 세포죽음을 보였다. 흥미롭게도 lactacytin은 정상세포인 HaCat 세포와 HGF-1 세포에서는 세포독성을 전혀 보이지 않았다. 그리고 lactacystin이 적용된 SCC25세포에서 핵 응축, DNA의 조각남, 사립체막전위와 proteasome 활성도의 감소, DNA 양의 감소, cytochrome c의 사립체에서의 세포질로의 유리, AIF와 DFF40 (CAD)의 핵으로의 이동, Bax의 증가, caspase-7, caspase-3, PARP, lamin A/C 그리고 DFF45 (ICAD)의 활성화 혹은 파괴와 같은 아주 다양한 세포자멸사 증거를 보였다. Flow cytometry 분석에서는 CDK 억제제인 $p21^{WAF1/CIP1}$와 $p27^{KIP1}$의 발현 증가와 관계있는 것으로 추정되어 지는 G1 세포주기 정지를 보였다. 이러한 결과는 lactacytin이 SCC25 세포에서 G1 세포주기정지와 proteasome, 사립체 및 caspase 경로의 연속반응을 통한 세포자멸사를 유도함을 명확하게 증명하고 있다. 이와 같은 세포주기 정지와 세포자멸사 유도능은 lactacytin이 사람혀편평상피세포암종의 새로운 치료전략으로서의 가능성을 제공한다고 생각한다.

• 동의생리병리학회지
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• 제20권1호
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• pp.163-170
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• 2006

Nardostachyos Rhizoma (N. Rhizoma) belonging to the family Valerianaceae has been anti-arrhythmic effect, and sedation to the central nerve and a smooth muscle. We reported that the water extract of N. Rhizoma induced apoptotic cell death and differentiation in human promyelocytic leukemia (HL-60) cells. Cytotoxicity of N. Rhizoma was detected only in HL-60 cells (IC50 is about 200 ${\mu}g/ml$). The cytotoxic activity of N. Rhizoma in HL-60 cells was increased in a dose-dependent manner. We used several measures of apoptosis to determine whether these processes were involved in N. Rhizoma-induced apoptotic cell death. The high-dose (200 ${\mu}g/ml$) treatment of N. Rhizoma to HL-60 cells showed cell shrinkage, cell membrane blobbing, apoptotic bodies, and the fragmentation of DNA, suggesting that these cells underwent apoptosis. Treatment of HL-60 cells with N. Rhizoma time-dependently induced activation of caspase-3, caspase-8, and caspase-9 and proteolytic cleavage of poly(ADP-ribose) polymerase. Also, we investigated the effect of N. Rhizoma on cellular differentiation and proliferation in HL-60 cells. Differentiation and proliferation of HL-60 cells was determined through expression of CD11b and CD14 surface antigens using flow cytometry and nitroblue tetrazolium (NBT) assay, and through analysis of cell cycle using propidium iodide assay, respectively. N. Rhizoma induced the differentiation of HL-60 at the low-dose (100 ${\mu}g/ml$) treatment, as shown by increased expression of differentiation surface antigen CD11b, but not CDl4 and increased reducing activity of NBT. When HL-60 cells were treated with N. Rhizoma at concentration of $50{\mu}g/ml\;and\;100{\mu}g/ml$, NBT-reducing activities induced approximately 1.5-fold and 20.0-fold as compared with the control. In contrast, HL-60 cells treated with the N. Rhizoma-ATRA combination showed markedly elevated levels of 26.3-fold at $50{\mu}g/ml$ N. Rhizoma-0.1 ${\mu}M$ ATRA combination and 27.5-fold at 50 ${\mu}g/ml$ N. Rhizoma-0.2 ${\mu}M$ ATRA combination than when treated with N. Rhizoma alone or ATRA alone. It may be that N. Rhizoma plays important roles in synergy with ATRA during differentiation of HL-60 cells. DNA flow-cytometry indicated that N. Rhizoma markedly induced a G1 phase arrest of HL-60 cells. N. Rhizoma-treated HL-60 cells increased the cell population in G1 phase from 32.71% to 42.26%, whereas cell population in G2/M and S phases decreased from 23.61% to 10.33% and from 37.78% to 33.98%, respectively. We examined the change in the $p21^{WAF1/Cip1}\;and\;p27^{Kip1}$ proteins, which are the CKIs related with the G1 phase arrest. The expression of the CDK inhibitor $p27^{Kip1},\;but\;not\;p21^{WAF1/Cip1}$ were markedly increased by N. Rhizoma. Taken together, these results demonstrated that N. Rhizoma induces apoptotic cell death through activation of caspase-3, and potently inhibits the proliferation of HL-60 cells via the G1 phase cell cycle arrest in association with $p27^{Kip1}$ and granulocytic differentiation induction .

• Tuberculosis and Respiratory Diseases
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• 제57권5호
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• pp.449-460
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• 2004

연구배경 : PS-341은 최근에 개발된 강력하고 특이적인 proteasome 억제제로서, 일부 암환자에 투여하여 좋은 성적이 보고되고 있다. Proteasome 억제제의 항암효과는 아포프토시스 유발 물질, 즉 p53, $p21^{WAF/CIP1}$, $p27^{KIP1}$, NF-${\kappa}B$, Bax, Bcl-2 등의 발현 증가와 관련이 있는 것으로 생각되고 있다. JNK와 GSK-$3{\beta}$도 아포프토시스에 관여하는 것으로 잘 알려져 있지만, PS-341에 의한 아포프토시스에서의 역할은 규명되지 못한 상태이다. 본 연구에서는 폐암세포주에서 PS-341에 의한 아포프토시스에서 JNK와 GSK-$3{\beta}$의 역할을 규명하고자 하였다. 방 법 : NCI-H157과 A549 폐암세포주를 실험에 사용하였다. 세포생존능은 MTT 방법으로 평가하였고, 아포프토시스는 PARP의 분해로 평가하였다. JNK의 활성도는 in vitro immuno complex kinase 방법과 내인성 c-Jun의 인산화로 측정하였다. 각종 단백의 발현은 Western 분석으로 평가하였다. JNK1과 GSK-$3{\beta}$의 과발현은 각각 plasmid vector와 adenovirus vector를 이용하였다. 결 과 : PS-341 처치로 아포프토시스에 의한 세포생존율의 감소가 관찰되었다. PS-341 처치로 JNK가 활성화되었고, c-Jun의 발현이 유도되었다. Dominant negative JNK1의 과발현 또는 SP600125 전치치로 JNK의 활성화를 차단하면 PS-341에 의한 아포프토시스가 억제되었다. PS-341 처리로 JNK 활성화에 의존적으로 caspase 3의 활성화가 유도되었다. Caspase 활성화의 차단으로도 PS-341에 의한 아포프토시스가 억제되었다. PS-341에 의해 Akt가 활성화되었고, Akt 활성화의 차단으로 PS-341에 의한 아포프토시스가 심화되었다. PS-341에 의한 Akt 활성화로 GSK-$3{\beta}$가 불활성화되었다. Constitutively active GSK-$3{\beta}$의 과발현으로 PS-341에 의한 아포프토시스가 심화되었고, dominant negative GSK-$3{\beta}$의 과발현으로 PS-341에 의한 아포프토시스가 감소되었다. Lithium chloride 전처치와 dominant negative GSK-$3{\beta}$의 과발현으로 PS-341에 의한 JNK의 활성화와 c-Jun의 발현 증가가 억제되었다. 결 론 : 폐암세포주에서 PS-341에 의한 아포프토시스는 JNK/caspase 경로가 관여하며, 이는 PI3K/Akt 경로를 통한 GSK-$3{\beta}$의 불활성화에 의해 억제되는 것으로 판단된다. 따라서 PS-341의 항암효과를 최대화하기 위해서는 PI3K/Akt 경로를 통한 GSK-$3{\beta}$의 불활성화를 차단하는 치료법이 병행되어야 할 것으로 판단된다.

• Asian-Australasian Journal of Animal Sciences
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• 제26권2호
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• pp.266-274
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• 2013

The effect of different phytogenic feed additives on reducing odorous compounds in swine was investigated using in vitro fermentation and analyzed their microbial communities. Soybean meal (1%) added with 0.1% different phytogenic feed additives (FA) were in vitro fermented using swine fecal slurries and anaerobically incubated for 12 and 24 h. The phytogenic FAs used were red ginseng barn powder (Panax ginseng C. A. Meyer, FA1), persimmon leaf powder (Diospyros virginiana L., FA2), ginkgo leaf powder (Ginkgo biloba L., FA3), and oregano lippia seed oil extract (Lippia graveolens Kunth, OL, FA4). Total gas production, pH, ammonianitrogen ($NH_3$-N), hydrogen sulfide ($H_2S$), nitrite-nitrogen ($NO_2{^-}$-N), nitrate-nitrogen ($NO_3{^-}$-N), sulfate (${SO_4}^{--}$), volatile fatty acids (VFA) and other metabolites concentration were determined. Microbial communities were also analyzed using 16S rRNA DGGE. Results showed that the pH values on all treatments increased as incubation time became longer except for FA4 where it decreased. Moreover, FA4 incubated for 12 and 24 h was not detected in $NH_3$-N and $H_2S$. Addition of FAs decreased (p<0.05) propionate production but increased (p<0.05) the total VFA production. Ten 16S rRNA DGGE bands were identified which ranged from 96 to 100% identity which were mostly isolated from the intestine. Similarity index showed three clearly different clusters: I (FA2 and FA3), II (Con and FA1), and III (FA4). Dominant bands which were identified closest to Eubacterium limosum (ATCC 8486T), Uncultured bacterium clone PF6641 and Streptococcus lutetiensis (CIP 106849T) were present only in the FA4 treatment group and were not found in other groups. FA4 had a different bacterial diversity compared to control and other treatments and thus explains having lowest odorous compounds. Addition of FA4 to an enriched protein feed source for growing swine may effectively reduce odorous compounds which are typically associated with swine production.

• 무역상무연구
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• 제60권
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• pp.3-26
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• 2013

Seller's obligation on the Delivery of Goods and Handing over the Documents are key elements in Contracts for the International Sale of Goods. The United Nations Convention on Contracts for the International Sale of Goods(CISG) has been entered into force on 1 January 1988 to create international certainty and uniformity in the law and to govern issues that arise in an international sale of goods transaction. The Incoterms were first published by the ICC in 1936 and were most recently revised in 2010. Incoterms 2010 are entering into force on 1 January 2011. The Incoterms focus on the seller's delivery obligations and reflect the principle that the risk of loss or damage to the goods passes from the seller to the buyer when the seller has fulfilled its obligations to deliver the goods. This study highlights basic rules covering seller's obligation of delivery of goods and handing over the documents under the Incoterms 2010 and the United Nations Convention and Contracts for the International Sale of Goods. In the second chapter, this study will provide analyses and compare these two legal systems in relation to the basic rules governing delivery of goods and passing of risks in contract of sale. This chapter evaluates the meaning of Article 31 and Article 67(1) and FOB, CFR, CIF & FCA, CPT, CIP terms of Incoterms 2010. Chapter Three will focus on handing over the documents. Article 30 CISG imposes the seller's primary obligations to deliver the goods and to hand over documents relating to them. Article 34 CISG supplements the seller's obligation in relation to documents by providing that the seller must hand over documents relating to the goods. In contrast, Article 58(1) CISG imposes on the buyer the obligation to pay only when it has received the goods or documents controlling their disposition. I reviewed only some of the documents relating to the goods are documents controlling their disposition. This chapter considers the meaning of the phrase "documents that control the disposition of the goods and do not control disposition of the goods." Finally, the fourth chapter will assess the meaning of rules of CISG and Incoterms 2010.

• 대한환경공학회지
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• 제39권3호
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• pp.155-163
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• 2017

최근 막여과 공정은 입자상 물질뿐만 아니라 Giardia, cryptosporidium과 같은 병원성미생물이 효과적으로 제거됨에 따라 국내외 적용성이 확대되는 추세이다. 본 연구는 광주광역시 D정수장의 원수 침전상등수의 처리를 위해 $120m^3/d$ 처리규모의 Pilot Plant (2014. 1월부터 운영)와 $2,500m^3/d$ 처리규모의 실증실험시설(2015. 1월부터 운영)의 막여과 공정을 운영하면서 최적 운전조건과 한계여과속도를 찾고자 하였다. 정밀여과막의 Pilot-test에서 얻어진 운전모드는 유입 1.0분, 여과 36.5분, 공기역세 0.9분, 역세 1.0분, 배출 1.0분이었으며, 이를 적용하여 실증실험을 하였다. 한계여과속도(critical flux)는 Pilot 운전에서 Flux 20, 40, 56 및 62 LMH일 때의 시간의 변화에 따른 막간차압의 증가식을 기반으로 $50L/m^2-h$로 산정(TMP 0.5bar 이내)되었다. 화학세정은 1차로 산세정 2회, 2차로 알칼리 세정을 하여 95%의 회복율을 얻었다. 이러한 적정 조건으로 운전한 결과 처리수의 탁도는 봄 여름 가을 겨울 평균 각각 0.028, 0.024, 0.026 및 0.028 NTU, 연평균 탁도는 0.026 NTU로, 탁도 제거율은 98.4%로 나타났다. 광주광역시 D정수장의 원수 침전상등수를 1일 $2,500m^3$ 처리하는 실증실험시설 운영을 통해 부유고형물 및 콜로이드성 물질을 제거하는데 탁월한 처리성능을 확인하였고, 일반적인 모래여과공정을 대체할 수 있는 적정운영기술이 확보되었다.

• 생명과학회지
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• 제28권9호
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• pp.1016-1021
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• 2018

C. difficile 톡신A에 의한 대장상피세포 자살과정은 위막성대장염(Pseudomembranous colitis)의 주요 원인으로 고려되고 있다. 톡신A는 활성산소 를 증가시켜 세포자살 신호를 유도한다. 또한 톡신A는 미세섬유나 미세소관과 같은 세포골격계 형성을 저해함으로써 자살을 유도한다고 알려져 있다. 하지만 톡신A가 야기하는 소화기 상피세포 자살경로는 아직 불분명하다. 본 연구에서는 소화관 상피세포의 성장과 분화 그리고 기능에 중요하다고 알려져 온 Wnt 신호경로에 대한 톡신A의 영향을 확인해보았다. 이를 위해 비암화-인간대장세포주(NCM460)에 톡신A를 처치하고 Wnt 신호 분자들의 변화를 추적하였다. 또한 톡신A를 주입한 생쥐의 회장 상피세포 속 Wnt 신호경로 변화도 평가하였다. 인간 대장상피세포에서 톡신A는 Wnt 경로의 핵심 신호분자인 ${\beta}$-catenin 단백질의 양을 빠르게 감소시켰다. 이 현상은 생쥐 회장 상피세포에서도 동일하게 확인되었다. 연구자 등은 톡신A가 $GSK3{\beta}$ 활성형 인산화(Thr390)를 증가시킴도 확인하였다. 이는 톡신A가 $GSK3{\beta}$의 활성을 높여서 ${\beta}$-catenin의 인산화시키고 이를 통해 단백질 분해 과정이 촉진되었음을 보여준다. 이 결과들을 종합하면, 톡신A에 의한 소화관 상피세포 자살과정이 상피세포의 성장과 자살을 조절하는 Wnt 신호경로 차단과 밀접하게 연관되어 있음을 보여준다.

• Journal of Microbiology and Biotechnology
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• 제24권12호
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• pp.1654-1663
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• 2014

To examine the effect of tumor suppressor protein p53 on the antitumor activity of 2-methoxyestradiol (2-MeO-$E_2$), 2-MeO-$E_2$-induced cell cycle changes and apoptotic events were compared between the human colon carcinoma cell lines HCT116 ($p53^{+/+}$) and HCT116 ($p53^{-/-}$). When both cell types were exposed to 2-MeO-$E_2$, a reduction in the cell viability and an enhancement in the proportions of $G_2/M$ cells and apoptotic sub-$G_1$ cells commonly occurred dose-dependently. These 2-MeO-$E_2$-induced cellular changes, except for $G_2/M$ arrest, appeared to be more apparent in the presence of p53. Immunofluorescence microscopic analysis using anti-${\alpha}$-tubulin and anti-lamin B2 antibodies revealed that after 2-MeO-$E_2$ treatment, impaired mitotic spindle network and prometaphase arrest occurred similarly in both cell types. Following 2-MeO-$E_2$ treatment, only HCT116 ($p53^{+/+}$) cells exhibited an enhancement in the levels of p53, p-p53 (Ser-15), $p21^{WAF1/CIP1}$, and Bax; however, the Bak level remained relatively constant in both cell types, and the Bcl-2 level decreased only in HCT116 ($p53^{+/+}$) cells. Additionally, mitochondrial apoptotic events, including the activation of Bak and Bax, loss of ${\Delta}{\psi}m$, activation of caspase-9 and -3, and cleavage of lamin A/C, were more dominantly induced in the presence of p53. The Bak-specific and Bax-specific siRNA approaches confirmed the necessity of both Bak and Bax activations for the 2-MeO-$E_2$-induced apoptosis in HCT116 cells. These results show that among 2-MeO-$E_2$-induced apoptotic events, including prometaphase arrest, up-regulation of Bax level, down-regulation of Bcl-2 level, activation of both Bak and Bax, and mitochondria-dependent caspase activation, the modulation of Bax and Bcl-2 levels is the target of the pro-apoptotic action of p53.

• 한국동물위생학회지
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• 제23권1호
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• pp.45-59
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• 2000

The present study was conducted to investigate the epidemiological properties of salmonellosis of poultry farms in Kyongbuk province during the relied from November 1998 to November 1999 And antibiotic susceptibility and biochemical characteristics of 120 Salmonella cultures isolated from chicken samples were also investigated. The results obtained through this study were summarized as follows, 1. Among 667,200 chickens of 31 flocks in 17 layer farms and 9 broiler farms, 61,350 chickens of 31 flocks were died with salmonellosis. The death rate of 25 farms varied from 0.1% to 75.0%, and the average death rate was 9.2%. 2. According to etiological agents, fowl typhoid was shown the most predominant outbreak among the salmonellosis during a year, which accounted for 8R.0%(22/25) of the total case 3. The serotypes of 120 Salmonella isolates were identified as 7 strains(5.8%) of S pullorum, 10 strains(8.4%) of S typhimurium and 103 strains(85.8%) of S gallinarum. 4. Most outbreak of fowl typhoid were prevalent on the layer chicken farms(77.1%), and the summer season(45.5%) also appeared the most hazardous season during the year. 5. It seemed that the Hyline breed(70.6%) was the most susceptible among the layer chicken breeds, and followed by Isabrown(23.5%), Tetra(5.9%) in order. 6. In layers, 76.4% of fowl typhoid occurred commonly from 14 to 40 weeks including the early laying peroid, but in broiler farms, all cases was outbreak within first second weeks. 7 All the strains of S pullorum were resistant to lincomycin(Lm), penicillin(Pm), and steptomycin(Sm), but sensitive to amikacin(Ak), ampicillin(Am), cephalothin(Ce), ciprofloxacin (CiP), chloramphenicol(Cm), colistin(Co), enrofloxacin(Enr), furazolidone(Fu), gentamicin(Gm), kanamycin(Km), neomycin(Nm), polymyxin(Po), and teracycline(Tc). All the strains of S typhimurium were resistant to Lm(100%), Pm(100%), Po(90%), and Sm(90%), but were sensitive to Ak, Am, Ce, CiP, Cm, Co, Enr, Fu, Gm, Km, Nm, and Tc. 8. Minimum inhibitory concentration(MIC) of 103 strains of S gallinarum were also evaluated and their patterns were much more variable than others. All the strains of S gallinarum were sensitive to Ak, Am, Ce, Cip, Cm, Enr, Fu, Km, and Nm, but resistant to Lm(100%), and Sm(100%), 99(96.1%) to Co, 83(80.6%) to Pm, and 83(80.6%) to Po, 55(53.4%) to Gm, and 33(32.0%) to Tc. 9. The multiple drug resistance patterns of 120 Salmonella strains were CoLmPmPo Sm pattern(34.2%), CoGmLmPmPoSmTc(20.8%), CoGmLmPmPoSm(13.3%), CoGmLmSm(7.5%), LmPmSm(7.5%), LmPm(6.7%), CoLmSmTc(3.3%),, CoGmLmSmTc(1.7%), GmLmSmTc(1.7%), CoGmLmPoSm(0.1%), LmPmPo(0.1%), CoLm Sm(0.1%), and LmSm(0.1%), in order.

• 상하수도학회지
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• 제21권5호
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• pp.559-569
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• 2007

The purposes of this study were to find the main foulant of membrane and the optimal chemical cleaning method for MF(microfiltration) drinking water treatment system using D dam water as water source. The MF pilot plant which can treat maximum $500m^3/d$ consisted of 3 racks and was operated for 10 months under various operation conditions. After 10 months operation, $1^{st}$ and $2^{nd}$ rack of membrane pilot plant system were cleaned chemically and the degree of the restoration of the fouled membrane in terms of the pure water flux was detemnined. Inorganic compounds which contained in chemical cleaning waste was analyzed by Inductively Coupled Plasma (ICP). One membrane module for 3rd rack was disjointed and membrane fouling materials, especially inorganic compounds were investigated by Electron Probe Microanlysis (EPMA) to elucidate the reason of TMP increase. And also, the various chemical reagents (1N HCl or $H_2SO_4$, oxalic acid as acid and 0.3% NaOCl as alkali) were tested by combination of acid and alkali to determine the optimal chemical cleaning method for the MF system using micro-modules manufactured using the disjointed module. It was verified that the inside and outside of membrane module was colorized with black. As a result of the quantitative and semi-qualitative analysis of membrane foulant by ICP, most of inorganic foulant was manganese which is hard to remove by inorganic acid such as HCI. Especially, it was observed by EPMA that Mn was attached more seriously in inside surface of membrane than in outside surface of that. It was supposed that Mn fouling in inside surface of membrane might be caused by the oxidation of soluble manganese (Mn(II)) to insoluble manganese ($MnO_2$) by chlorine containing in backwashing water. The optimal cleaning method for the removal of manganese fouling was consecutive cleaning with the mixture of 1N HCl and 1% of oxalic acid, 0.3% NaOCl, and 1N HCl showing 91% of the restoration of the fouled membrane.