• 한국약용작물학회지
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• 제18권2호
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• pp.79-85
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• 2010

Enhancement effect of ultrasonification process on UV-protection and skin-whitening activities using Centella asiatica L. Urban extract was investigated. Cytotoxicity of the extracts measured on human skin fibroblast cells, CCD-986sk, and then, ultrasonification associated extracts showed 5~9% lower cytotoxicity then normal crude extracts on 1.0 mg/$m{\ell}$ of highest sample concentration. The associated extrats showed highest inhibition activity of hyaluronidase on 1.0 mg/$m{\ell}$ of concentration as 54.2%. Also, the associated extract reduced expression of MMP-1 on UV-irradiated CCD-986sk cells down to 100.2% from 136.1%, and revealed high inhibitory potency on tyrosinase as 74.6% by adding 1.0 mg/$m{\ell}$ of concentration. Ultrasonification associated extract showed strong inhibition effect of melanin production on Clone M-3 cells as 84.2% by adding 1.0 mg/$m{\ell}$ of concentration. From the preliminary observations, we considered that the extracts from C. asiatica could be potent natural materials for skin-whitening and anti-aging agent, and could enhance the activities by ultrasonification process.

• 대한바이러스학회지
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• 제29권4호
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• pp.235-245
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• 1999

HIV-1 Tat, a strong transactivator, is essential for the HIV-1 replication and AIDS progression. The Tat function is markedly inhibited by human p53 anti-oncogene. However, the detail mechanism has not yet been clearly revealed. In our previous report, we have addressed that p53 is unlikely to interact directly with HIV-1 Tat. In the consecutive experiments, Tat-phosphorylation was found to increase in proportional to the amounts of transfected p53. This work was initiated to identify the signaling factor that is involved in the p53-mediated Tat suppression. Several protein kinases were tested for the phosphorylation of Tat, and we found that PKR is likely to be involved in the p53-mediated Tat suppression. PKR was co-immunoprecipitated by anti-Tat antibody in the Tat-expressing Jurkat cell lysates only when the cells were transfected by p53, indicating that PKR-Tat interaction depends on the p53 activity. The interaction seems to result in PKR-mediated Tat-phosphorylation. Tat function was not blocked by p53 when co-transfected trasiently with antisense-PKR. We have generated PKR-knock out Jurkat cell clone. The PKR defective Jurkat cells didn't show the p53-mediated Tat suppression. These data indicate that p53-mediated Tat suppression is strongly associated with PKR. PKR-mediated Tat phosphorylation experiments are now under investigation by kinase assay and co-immunoprecipitation in the presence or absence of p53.

• 생약학회지
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• 제32권2호통권125호
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• pp.163-167
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• 2001

Prunella vulgaris L. aqua-acupuncture solution (PVAS) was tested for cancer chemopreventive activity using chemoprevention-associated biochemical end points. The following effects were measured. : (a) inhibition of 7,12-dimethylbenz[a]anthracene (DMBA)-induced cytochrome P4501A1 activity (b) inhibition of $[^3H]B[a]P-DNA$ binding (c) inhibition of phorbol 12-myristate 13-acetate (TPA)-induced free radical formation in HL-60 cells (d) inhibition of polyamine metabolism. PVAS inhibited cytochrome P4501A1-mediated ethoxyresorufin-O-deethylase activity. The binding of $[^3H]B[a]P$ metabolites to DNA of NCTC-clone 1469 cells was inhibited significantly by PVAS. There is 22% inhibition of TPA-induced free radical formation in human leukemic cells with 5 mg/ml PVAS. Proliferation of Acanthamoeba castellanii was inhibited by PAVS at concentration of 30 mg/ml. PAVS positive in these assays may inhibit the carcinogenesis process and is considered very promising cancer-preventing agent because of its multiple activities.

• 한국약용작물학회지
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• 제19권3호
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• pp.191-197
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• 2011

In this study, we investigated antioxidant activities and whitening effects of Acer mono sap by encapsulation of nanoparticles. Acer mono sap was through ultra high pressure process and then encapsulated by lecithin. Nano-encapsulated The nanoparticles of Acer mono sap showed highest free radical scavengering effect as 89.7% in adding sample (1.0 mg/ml), compared to sap of non-encapsulation. It was showed strong inhibition effect on melanin production test by Clone M-3 cells as 47.8%. High inhibitory of tyrosinase was also measured as 85.8% by adding lecithin nano-particle of 1.0 mg/ml. The nano-particles also showed 14.8% of low cytotoxicity against human normal fibroblast cells in adding 1.0 mg/ml of the highest concentration. These results indicate that Acer mono sap may be a source of cosmetic agents capable of improving whitening effect and antioxidant activites.

• 대한핵의학회지
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• 제37권1호
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• pp.43-52
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• 2003

Microarray technology allows the simultaneous analysis of gene expression patterns of thousands of genes, in a systematic fashion, under a similar set of experimental conditions, thus making the data highly comparable. In some cases arrays are used simply as a primary screen loading to downstream molecular characterization of individual gene candidates. In other cases, the goal of expression profiling is to begin to identify complex regulatory networks underlying developmental processes and disease states. Microarrays were originally used with ceil lines or other simple model systems. More recently, microarrays have been used in the analysis of more complex biological tissues including neural systems and the brain. The application of cDNA arrays in neuropsychiatry has lagged behind other fields for a number of reasons. These include a requirement for a large amount of input probe RNA In fluorescent-glass based array systems and the cellular complexity introduced by multicellular brain and neural tissues. An additional factor that impacts the general use of microarrays in neuropsychiatry is the lack of availability of sequenced clone sets from model systems. While human cDNA clones have been widely available, high qualify rat, mouse, and drosophilae, among others are just becoming widely available. A final factor in the application of cDNA microarrays in neuropsychiatry is cost of commercial arrays. As academic microarray facilitates become more commonplace custom made arrays will become more widely available at a lower cost allowing more widespread applications. in summary, microarray technology is rapidly having an impact on many areas of biomedical research. Radioisotope-nylon based microarrays offer alternatives that may in some cases be more sensitive, flexible, inexpensive, and universal as compared to other array formats, such as fluorescent-glass arrays. In some situations of limited RNA or exotic species, radioactive membrane microarrays may be the most practical experimental approach in studying psychiatric and neurodegenerative disorders, and other complex questions in the brain.

• Journal of Ginseng Research
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• 제43권3호
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• pp.452-459
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• 2019

Background: Ginsenoside compound K(C-K), a major metabolite of ginsenoside, exhibits anticancer activity in various cancer cells and animal models. A cell signaling study has shown that C-K inhibited nuclear factor-kappa B ($NF-{\kappa}B$) pathway in human astroglial cells and liver cancer cells. However, the molecular targets of C-K and the initiating events were not elucidated. Methods: Interaction between C-K and Annexin A2 was determined by molecular docking and thermal shift assay. HepG2 cells were treated with C-K, followed by a luciferase reporter assay for $NF-{\kappa}B$, immunofluorescence imaging for the subcellular localization of Annexin A2 and $NF-{\kappa}B$ p50 subunit, coimmunoprecipitation of Annexin A2 and $NF-{\kappa}B$ p50 subunit, and both cell viability assay and plate clone formation assay to determine the cell viability. Results: Both molecular docking and thermal shift assay positively confirmed the interaction between Annexin A2 and C-K. This interaction prevented the interaction between Annexin A2 and $NF-{\kappa}B$ p50 subunit and their nuclear colocalization, which attenuated the activation of $NF-{\kappa}B$ and the expression of its downstream genes, followed by the activation of caspase 9 and 3. In addition, the overexpression of Annexin A2-K320A, a C-K binding-deficient mutant of Annexin A2, rendered cells to resist C-K treatment, indicating that C-K exerts its cytotoxic activity mainly by targeting Annexin A2. Conclusion: This study for the first time revealed a cellular target of C-K and the molecular mechanism for its anticancer activity.

• Journal of Microbiology and Biotechnology
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• 제31권1호
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• pp.144-153
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• 2021

Organophosphorus nerve agents (OPNAs), including both G- and V-type nerve agents such as sarin, soman, tabun and VX, are extremely neurotoxic organophosphorus compounds. Catalytic bioscavengers capable of hydrolyzing OPNAs are under development because of the low protective effects and adverse side effects of chemical antidotes to OPNA poisoning. However, these bioscavengers have certain limitations for practical application, including low catalytic activity and narrow specificity. In this study, we generated a fusion-hybrid form of engineered recombinant human paraoxonase 1 (rePON1) and bacterial organophosphorus hydrolase (OPH), referred to as GV-hybrids, using a flexible linker to develop more promising catalytic bioscavengers against a broad range of OPNAs. These GV-hybrids were able to synergistically hydrolyze both G-type OPNA analogs (paraoxon: 1.7 ~ 193.7-fold, p-nitrophenyl diphenyl phosphate (PNPDPP): 2.3 ~ 33.0-fold and diisopropyl fluorophosphates (DFP): 1.4 ~ 22.8-fold) and V-type OPNA analogs (demeton-S-methyl (DSM): 1.9 ~ 34.6-fold and malathion: 1.1 ~ 4.2-fold above) better than their individual enzyme forms. Among the GV-hybrid clones, the GV7 clone showed remarkable improvements in the catalytic activity toward both G-type OPNA analogs (kcat/Km (106 M-1 min-1): 59.8 ± 0.06 (paraoxon), 5.2 ± 0.02 (PNPDPP) and 47.0 ± 6.0 (DFP)) and V-type OPNA analogs (kcat/Km (M-1 min-1): 504.3 ± 48.5 (DSM) and 1324.0 ± 47.5 (malathion)). In conclusion, we developed GV-hybrid forms of rePON1 and bacterial OPH mutants as effective and suitable catalytic bioscavengers to hydrolyze a broad range of OPNA analogs.

• Pediatric Infection and Vaccine
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• 제6권2호
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• pp.253-260
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• 1999

목 적 : human CMV가 생성하는 분자량 72,000 dalton의 immediate early protein(p72)과 반응하는 단세포군 항체를 국내에서 분리된 CMV 야생주로 직접 제작하여 진단에 사용해 보기 위한 것이다. 방 법 : 정상 인체 섬유아세포(Foreskin 유래)를 10% 우태아혈청이 함유된 Eagle' s minimal essential medium(MEM)에서 배양하여 사용하였으며 마우스 골수종세포는 P3X63 Ag8.653(ATCC, Mary land USA)을 $5{\times}10^5/ml$ 세포수를 유지하도록 배양하였다. 사용된 항원은 선천성 CMV 감염아의 소변으로부터 검출한 CMV(KJHJ90)를 대량 증식시켜 사용하였다. 생후 6~8주된 Balb/c마우스의 복강내에 CMV 항원을 1주일 간격으로 4회 주사한 후 면역시킨 마우스 비장에서 얻은 임파구와 형질세포종세포로 최적발육기의 P3X63-Ag8.653을 사용하여 융합하였다. 융합세포를 배양한 후 간접면역형광항체법을 이용하여 항체가가 높은 것을 골랐다. 결 과 : 생산된 28종의 단세포군항체 중 그림 1과 같이 LPC12와 LPC23클론이 AD169에 감염된 세포의 핵에 특히 강하게 반응하였다. 정제된 AD169를 SDS-PAGE한 후 Western blotting을 하여 생산된 단세포군항체가 반응하는 항원의 분자량을 확인하였으며 그 중 LPC12와 LPC23클론에서 생성된 단세포군항체는 약 72KDA의 항원과 특이적으로 가장 강하게 반응하였다. 결 론 : 본 연구에서 제작된 LPC12와 LPC23 클론에서 생성된 단세포군 항체를 이용하여 선천성 CMV감염으로 확인된 소아의 소변을 이용하여 CMV AD 169 주 유래 항체(P63-27)과 비교한 DEAFF 검사상 동일한 결과를 확인한 바 LPC12와 LPC 23 클론 항체는 AD 169(P63-27)과 같이 CMV 감염의 조기진단에 사용할 수 있을 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권12호
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• pp.1649-1659
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• 2012

Crosses between Korean and Landrace pigs have revealed a large quantitative trait loci (QTL) region for fat deposition in a region (89 cM) of porcine chromosome 4 (SSC4). To more finely map this QTL region and identify candidate genes for this trait, comparative mapping of pig and human chromosomes was performed in the present study. A region in the human genome that corresponds to the porcine QTL region was identified in HSA1q21. Furthermore, the LMNA gene, which is tightly associated with fat augmentation in humans, was localized to this region. Radiation hybrid (RH) mapping using a Sus scrofa RH panel localized LMNA to a region of 90.3 cM in the porcine genome, distinct from microsatellite marker S0214 (87.3 cM). Two-point analysis showed that LMNA was linked to S0214, SW1996, and S0073 on SSC4 with logarithm (base 10) of odds scores of 20.98, 17.78, and 16.73, respectively. To clone the porcine LMNA gene and to delineate the genomic structure and sequences, including the 3'untranslated region (UTR), rapid amplification of cDNA ends was performed. The coding sequence of porcine LMNA consisted of 1,719 bp, flanked by a 5'UTR and a 3'UTR. Two synonymous single nucleotide polymorphisms (SNPs) were identified in exons 3 and 7. Association tests showed that the SNP located in exon 3 (A193A) was significantly associated with weight at 30 wks (p<0.01) and crude fat content (p<0.05). This association suggests that SNPs located in LMNA could be used for marker-assisted selection in pigs.

• Asian-Australasian Journal of Animal Sciences
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• 제23권5호
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• pp.640-648
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• 2010

Insulin-responsive glucose transporter 4 (GLUT4) is a member of the glucose transporter family and mainly presents in skeletal muscle and adipose tissue. To clarify the molecular structure of porcine GLUT4, RACE was used to clone its cDNA. Several cDNA clones corresponding to different regions of GLUT4 were obtained by amplifying reverse-transcriptase products of total RNA extracted from Landrace porcine skeletal muscles. Nucleotide sequence analysis of the cDNA clones revealed that porcine GLUT4 cDNA was composed of 2,491 base pairs with a coding region of 509 amino acids. The deduced amino acid sequence was over 90% identical to human, rabbit and cattle GLUT4. The tissue distribution of GLUT4 was also examined by Real-time RT-PCR. The mRNA expression abundance of GLUT4 was heart>liver, skeletal muscle and brain>lung, kidney and intestine. The developmental expression of GLUT4 and insulin receptor (IR) was also examined by Real-time RT-PCR using total RNA extracted from longissimus dorsi (LM), semimembranosus (SM), and semitendinosus (SD) muscle of Landrace at the age of 1, 7, 30, 60 and 90 d. It was shown that there was significant difference in the mRNA expression level of GLUT4 in skeletal muscles of Landrace at different ages (p<0.05). The mRNA expression level of IR also showed significant difference at different ages (p<0.05). The developmental change in the mRNA expression abundance of GLUT4 was similar to that in IR, and both showed a higher level at birth and 30 d than at other ages. However, there was no significant tissue difference in the mRNA expression of GLUT4 or IR (p>0.05). These results showed that the nucleotide sequence of the cDNA clones was highly identical with human, rabbit and cattle GLUT4 and the developmental change of GLUT4 mRNA in skeletal muscles was similar to that of IR, suggesting that porcine GLUT4 might be an insulin-responsive glucose transporter. Moreover, the tissue distribution of GLUT4 mRNA showed that GLUT4 might be an important nutritional transporter in porcine skeletal muscles.