• Journal of Microbiology and Biotechnology
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• 제18권2호
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• pp.328-333
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• 2008

A series of pep tides binding to the HCV NS5B polymerase was selected from phage display peptide libraries. A conserved motif of Ser-Arg-X-Arg/Leu was identified among the selected peptides, and Pep2 (Trp-Ser-Arg-Pro-Arg-Ser-Leu) was chosen for further characterization. The binding of Pep2 to HCV NS5B in vivo was shown by a yeast two-hybrid assay and by subcellular colocalization analysis using immunofluorescence confocal microscopy. The in vitro interaction was also confirmed by GST pulldown assay. The replication of the HCV 1b subgenomic replicon was efficiently inhibited by the presence of the peptide. By using a subtractive biopanning against Pep2, the binding site of the peptide was mapped at the pocket of Pro388 to Pro391 in the thumb subdomain of the polymerase. A yeast two-hybrid analysis using Pro388Ala and Pro391Ala mutants of NS5B confirmed the binding.

• Molecules and Cells
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• 제21권3호
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• pp.330-336
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• 2006

Since its identification in 1989, hepatitis C virus has been the subject of extensive research. The biology of the virus and the development of antiviral drugs are closely related. The RNA polymerase activity of nonstructural protein 5B was first demonstrated in 1996. NS5B is believed to localize to the perinuclear region, forming a replicase complex with other viral proteins. It has a typical polymerase structure with thumb, palm, and finger domains encircling the active site. A de novo replication initiation mechanism has been suggested. To date, many small molecule inhibitors are known including nucleoside analogues, non-nucleoside analogues, and pyrophosphate mimics. NS5B interacts with other viral proteins such as core, NS3, 4A, 4B, and 5A. The helicase activity of NS3 seems necessary for RNA strand unwinding during replication, with other nonstructural proteins performing modulatory roles. Cellular proteins interacting with NS5B include VAMP-associated proteins, heIF4AII, hPLIC1, nucleolin, PRK2, ${\alpha}$-actinin, and p68 helicase. The interactions of NS5B with these proteins might play roles in cellular trafficking, signal transduction, and RNA polymerization, as well as the regulation of replication/translation processes.

• 한국토양비료학회지
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• 제8권2호
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• pp.69-80
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• 1975

수도(水稻)의 수량(收量)과 여러가지 질소효율간, 질소효율 상호간(相互間), 효율과 흡수량간(吸收量間)의 관계(關係)를 설정(設定)하고 3개(個) 연간(年間)의 3요소시험(要素試驗) (30~50개지역(個地域)) 결과자료(結果資料)로 검토(檢討)하였다. 설정(設定)된 상호관계(相互關係)는 고도유의상관(高度有意相關)을 보이므로 실험결과(實驗結果)에 잘 일치(一致)하였다. 시비하(施肥下)에서 다수성(多收性)은 시료(肥料)의 이용율(利用率)(Eu)을 증가(增加)시켜 일차적(一次的)으로 질소흡수(窒素吸收)를 증가(增加)시키고 흡수시료질소吸收肥料窒素(Nf) 효율(Ef) 및 시비효율(Fe)을 증가(增加)시키며 이차적(二次的)으로 질소효율(E)을 증가(增加)시키는데 의존(依存)한다. 흡수(吸收)된 토양질소(土壤窒素)(Ns) 효율(Es)은 Ef보다 E에 대(對)한 기여도(寄與度)가 컸으며 모든 질소효율은 동반질소(同伴窒素)의 흡수량(吸收量) 및 상대(相對)되는 다른 질소효율에 역상관(逆相關)을 보였다. Es와 Ef는 1. 감법(減法) 2. Cs (Cs=Ns/Ns+Nf) 대(對) E plotting 법(法)과 3. 표식비료(標識肥料)를 사용(使用)한 E-Cs 및 Y-Ns Plotting 법(法)이 있으며 Plotting 법(法)은 E-Es Cs+B 식(式) 또는 Y=Es Ns+Ef Nf식(式)을 사용(使用)하며 B=Ef Cf로 Ef Nf와 함께 주어진 조건하(條件下에서 상수(常數)로 본다. Es는 Ef보다 대부분(大部分)의 경우 (80%) 크며 포장간(圃場間)에 Es보다 Ef에 차이가 크며 Ef는 특히 비료(肥料)의 형태(形態)에 의존(依存)한다. 설정검토(設定檢討)된 상호관계(相互關係)는 다음과 같다. 1. Y=$Es{\cdot}Ns+Ef{\cdot}Nf$ (Y는 수량(收量)) 2. E=$Es{\cdot}Cs+Ef{\cdot}Cf(Cf=Nf/Ns/Nf)$ 3. E=b-aN, E=E, Es 또는 Ef이고 N=N, Ns 또는 Nf이다. (E=Y/N, N=Ns+Nf), b는 주어진 조건(條件)에서의 E의 이론적(理論的) 최대치(最大値)이고 a는 Y=EN 곡선(曲線)의 N=0에서의 접선(接線)의 기울기이다. 4. Fe=$Ef{\cdot}Eu$, Se=$Es{\cdot}Eu$ (Se는 토양유효질소의 종조생산효율) 5. E=$Se{\cdot}Cs/Eu+Fe{\cdot}Cf/Eu$ 6. Y=$Es{\cdot}Eu{\cdot}Sf+Ef{\cdot}Eu{\cdot}Fn$ 또는 Y=$Es{\cdot}Eu{\cdot}Ea{\cdot}Sn+Ef{\cdot}Eu{\cdot}Fn(Sf=Ea{\cdot}Sn$, Ea는 비료질소(肥料窒素)와 대등(對等)한 토양유효질소(Sf)를 전토양질소(全土壤窒素)(Sn)로 나눈 유효화율).

• 인터넷정보학회논문지
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• 제25권2호
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• pp.93-99
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• 2024

본 논문에서는 점프 항을 포함하는 이자율 기간구조 모형의 채권 가격을 결정하기 위하여 이토의 보조정리(Ito's Lemma)를 적용하여 채권가격편미분방정식(Partial Differential Bond Price Equation; PDBPE)을 유도한다. PDBPE으로부터, 지수함수에 대한 매클로린 급수 (Maclaurin series; MS)와 적률생성함수(moment-generating function; MGF)를 이용하여 채권 가격의 수치해(Numerical Solution; NS)를 구한다. 그리고 몬테 카르로 시뮬레이션(Monte Carlo Simulation; MCS) 기법을 이용하여 채권의 가격을 결정하기 위한 알고리즘을 제안하고, 시뮬레이션 과정을 통하여 채권의 가격을 결정한다. 수치적 분석을 이용한 채권 가격의 NS와 MCS를 이용하여 얻은 채권 가격의 결과를 비교하기 위하여, NS의 값과 MCS의 값의 비율인 상대오차(Relative Error; RE)를 구한다. 이로부터 얻은 RE가 약 2.2%보다 작음을 확인할 수 있고, 이것은 수치적 분석뿐만 아니라 제안한 알고리즘을 이용해도 채권의 가격을 매우 정확하게 예측할 수 있음을 의미한다. 또한, 지수함수에 대한 MS를 이용하여 얻은 채권 가격의 NS가 MGF를 적용하여 구한 채권 가격의 NS보다 상대적으로 오차가 작다는 것을 확인할 수 있다.

• 대한전자공학회논문지TC
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• 제37권1호
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• pp.88-97
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• 2000

초고속이면서 소요 메모리의 크기를 극소화한 IP 라우터용 Lookup 알고리즘을 제안하고 성능을 분석하였다. 메모리 크기가 작으므로 고속/고가의 SRAM(10ns)을 사용할 수 있고, 구조가 간단하여 하드웨어로 구현 가능하였다. 본 장치는 1${\sim}$3회의 메모리 접근을 통해 Lookup이 가능하고, IPMA 사이트에서 구한 40,000개의 라우팅 정보를 이용하여 시뮬레이션한 결과 대략 ${\sim}$316KB의 포워딩 테이블용 메모리만이 소요된다. 이때 압축을 수행하는 옵셋 임계치는 8이다. ALTERA EPM7256시리즈에 100MHz 클럭을 이용하여 모사시험한 결과 10ns 접근속도를 가진 SRAM 기준으로 2회의 메모리 접근만으로 Lookup하는 경우 45ns의 접근시간이 소요되며, 3회의 메모리 접근이 필요한 경우는 ${\sim}$177ns의 접근시간이 소요된다.

• 한국분말재료학회지
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• 제5권2호
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• pp.89-97
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• 1998

Sintering behavior of nanostructured(NS) W-Cu powders prepared by mechanical alloying (MA) was investigated as a function of sintering temperature. MA NS W-2owt%Cu and W-3owt%Cu composite powders with the crystal size of 20-30 nm were annealed at 90$0^{\circ}C$, and thermal characteristics of those powders were investigated by DSC. Sintering behavior of MA NS W-Cu composite powders was investigated during the solid-state sintering and the Cu-liquid phase sintering. The new nanosintering phenonenon of MA W-Cu powders at solid-state sintering temperature was suggested to explain the W-grain growth in the inside of MA powders. The sintering densification of MA NS W-Cu powders was enhanced at Cu melting temperature by arrangement of MA powders, i.e., the first rearrangement of MA powders was occurred, and then the rearrangement of W-grains in the sintered parts was also took place during liquid-phase sintering, i.e., the second rearrangement was happened. Due to the double rearrangement process of MA NS W-Cu powders, the high sintered density with more than 96%o was obtained and the fine and high homogeneous state of W and Cu phases was achieved by sintering at 1200 $^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• 제33권6호
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• pp.788-796
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• 2023

Canine parvovirus type 2 (CPV-2) has high morbidity and mortality rates in canines. Nonstructural protein 1 (NS1) of CPV-2 has endonuclease activity, initiates viral DNA replication, and is highly conserved. Thus, it is a promising target for antiviral inhibitor development. We overexpressed a 41.9 kDa active recombinant endonuclease in Escherichia coli and designed a nicking assay using carboxyfluorescein and quencher-linked ssDNA as substrates. The optimal temperature and pH of the endonuclease were 37℃ and pH 7, respectively. Curcumin, bisdemethoxycurcumin, demethoxycurcumin, linoleic acid, tannic acid, and α-tocopherol inhibited CPV-2 NS1 endonuclease with IC₅₀ values of 0.29 to 8.03 µM. The extracted turmeric, yerba mate, and sesame cake suppressed CPV-2 NS1 endonuclease with IC₅₀ values of 1.48, 7.09, and 52.67 ㎍/ml, respectively. The binding affinity between curcumin, the strongest inhibitor, and CPV-2 NS1 endonuclease by molecular docking was -6.4 kcal/mol. Curcumin inhibited CPV-2 NS1 endonuclease via numerous hydrophobic interactions and two hydrogen bonds with Lys97 and Pro111 in the allosteric site. These results suggest that adding curcuminoids, linoleic acid, tannic acid, α-tocopherol, extracted turmeric, sesame cake, and yerba to the diet could prevent CPV-2 infection.

• 한국미생물·생명공학회지
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• 제36권4호
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• pp.353-359
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• 2008

Hepatitis C virus (HCV) is known as the causative agent of blood transmitted hepatitis. Two viral proteins, E2 and NS5A, are known to exert interferon resistance of HCV via PKR pathway. Here, we report a third protein, the RNA-dependent RNA polymerase (NS5B) of HCV, induced interferon resistance inhibiting p56 pathway. p56 was shown to interact with p48 subunit of eukaryotic initiation factor 3 (eIF3). This interaction inhibited formation of ternary complex in translation initiation. Using dual reporter assay system, we observed that the translation decreased when interferon alpha was added to the culture. But, in the presence of HCV NS5B, the translation partly recovered. NS5B and p48 subunit of eIF3 were shown to interact. This interaction seems to inhibit the interaction between p48 and p56. This is the first report that a virus exerts interferon resistance via p56 pathway.

• Journal of Ginseng Research
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• 제43권2호
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• pp.291-299
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• 2019

Background: Ginsenosides of Korean Red Ginseng extracts (RGE) and its saponin components suppress secretion of inflammasome-mediating cytokines, whereas the nonsaponin fraction (NS) of RGE oppositely stimulates cytokine secretion. Although direct exposure of NS to macrophages in mice induces cytokine production, oral administration of NS has not been studied in inflammasome-related disease in animal models. Methods: Mice were fed RGE or NS for 7 days and then developed peritonitis. Peritoneal cytokines were measured, and peritoneal exudate cells (PECs) were collected to assay expression levels of a set of toll-like receptors (TLRs) and cytokines in response to NS ingestion. In addition, the role of intestinal bacteria in NS-fed mice was assessed. The effect of preexposure to NS in bone marrow-derived macrophages (BMDMs) on cytokine production was further confirmed. Results: NS ingestion attenuated secretion of peritoneal cytokines resulting from peritonitis. In addition, the isolated PECs from NS-fed mice presented lower TLR transcription levels than PECs from control diet-fed mice. BMDMs treated with NS showed downregulation of TLR4 mRNA and protein expression, which was mediated by the $TLR4-MyD88-NF{\kappa}B$ signal pathway. BMDMs pretreated with NS produced less cytokines in response to TLR4 ligands. Conclusion: NS administration directly inhibits TLR4 expression in inflammatory cells such as macrophages, thereby reducing secretion of cytokines during peritonitis.

• 대한바이러스학회지
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• 제27권2호
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• pp.197-207
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• 1997

To investigate the NS4 region of JEV, NS4 cDNA of K94P05 (JEV strain isolated from Korea in 1994) was amplified by RT-PCR and analyzed by sequencing PCR product. Genomic size of NS4 was 1212bp and nucleotide sequence was compared with that of other JEV strains. Nucleotide homology between JaOAr582 and K94P05 was 91.1% and that between Beijing and K94P05 was 89.8%, respectively. But the nucleotide sequence of E region of JaOAr582 and K94P05 showed 97.0% homology and that of Beijing and K94P05 did 95.8% homology. NS4 protein was expressed as a form of fusion protein by a prokaryotic expression system. The induced fusion product showed a lower molecular weight than predicted size and remained insoluble. The NS4 protein might be cleavaged by E. coli protease. Concluding above results, high hydrophobicity of the NS4 protein supported the fact that this protein played a role as a membrane component and the poor nucleotide sequence conservativity among JEV strains suggested that this region might be important to adapt each viral growth environment.