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Tumor Surpressor Gene Therany, and Natural Product with Vectors[Aoenouirus, Aoenn associated virus] in Human Papilloma virus (HPV[Human papilloma virus]유래 바이러스 벡터[Adenovirus, Adeno associated virus]를 이용한 암 억제유전자치료법과 자연산물에서의 암 억제 효과)

  • 천병수;노민석;유종수;김준명
    • KSBB Journal
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    • v.16 no.6
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    • pp.579-591
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    • 2001
  • The cell growth inhibitor effect of cervical cancer cells was investigated by liposome mediated transfection (pRcCMVp53/lipofectin) and by transfection using adenovirus (AdCMVp57). The papilloma virus cancer cell lines we used in this study were HPV16 positive, having inhibiter gene, wild p53 gene, CaSki, SiHa, HPV18 positive HeLa, HeLaS3 and HPV negative C33A, HT3. LacZ gene of E.coli was used as the marker gene for the transfection efficiency. The effect on the inhibition of tumor cell growth was measured by cell count and cell viability though ELISA analysis and MTT assay. The inhibition of tumor cell growth was confirmed by measuring each assay for six days, comparing with the normal control cell growth. The cell growth of cervical cancer calls by transfection was significantly reduced and showed tittle differences among the cell lines. To eliminate the potential problem of Ad(adenovirus) contamination during rAAV production, rAAV can be produced by a triple transfection of vector plasmic, packaging plasmid, and adenovirus helper plasmid. To examine the helper functions of Ad plasmids on the production of rAAV vector, we carried out cotransfection of three plasmids, AAV vector, packaging construct, and Ad helper plasmids. The optimized transfection condition for calcium phosphate method is 25ug of total DNA per 10-cm-diameter plate of 293 cell. We found that rAAV yields peaked at 48hr after Ad infection. The titer of rAAV was measured by the dot blot analysis to measure the number of particles/ml based on the quantification of viral DNA. Recent1y, Kombucha(fungi) was identified as a very potent antileukefic agent. In the present study, effect of natural toxin(plankton) and Kombucha is PSP(GTXI-3, neoSTX), on various MTT assay cervical cancer cell line. Toxin(GTX 1-3, neoSTX) also inhibited the proliferation in primary cervical cancer calls in a dose-dependent toxin concentration. These results showed that toxin was very potent in inhibiting the proliferation of cervical cancer calls in vitro. Toxins and Kombuoha exhibited a dose dependent inhibition of cellular proliferation in cancer cell line.

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Expression of $O^6$-methylguanine-DNA methyltransferase and Sensitivity to Anticancer Alkylating Agents in Human Cancer Cells (사람 암세포에서의 $O^6$-methylguanine-DNA methyltransferase의 발현과 알킬화 항암제에 대한 감수성)

  • 오혜영;정해관;한의식;정성철;허옥순;손수정;김영미;홍성렬;이향우
    • Biomolecules & Therapeutics
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    • v.3 no.2
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    • pp.122-131
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    • 1995
  • Five human cancer cell lines (HeLa S3, Hep 3B, KATO III, Hs 683, HeLa MR) and one human normal cell line (WI-38) were examined cell viability, northern blot analysis, western blot analysis, and in situ hybridization for the expression $O_{6}$ -methylguanine-DNAmethyltransferase (MGMT), which can repair $O_{6}$ -methylguanine produced in DNA by alkylating agents. In cell viability test, the lethal sensitivities of each strain against anti-tumor drug N,N-bis(2-chloroethyl)- N-nitrosourea (BCNU) were counted, and both BCNU treated and untreated cell extracts were examined for their MGMT inducibility by RNA dot blot analysis. Cell lines did not show MGMT induction by BCNU pretreatment. Tlle MGMT activity was assayed by measuring the $^3$H radioactivity transferred from the substrate DNA containing [methyl-$^3$H)-O$_{6}$ -methylguanine to acceptor molecules in the cell extracts. Extracts from the majority of tumor strains and normal cells contained substantial MGMT activity of varying degree, while the known Mer$^{[-10]}$ cell (lacked or severely depleted in MGMT activity) Hela MR, and Hs 683 (proved to be Mer$^{[-10]}$ ) were much more sensitive to BCNU than the rest of tumor strains, as measured by cell viability test. Overall results above, KATO III showed the highest expression level of MGMT among the strains examined. Furthermore, with all the tumor and normal strains tested, a good correlation was observed between MGMT expression and cellular resistance to BCNU. The varying levels of expression of MGMT in human cancer cells found in this study should provide a molecular basis for MGMT expression among tumor strains from different tissue origin, the information of antitumor agents selection for chemotherapy of cancers.

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Improvement of ${\beta}-glucosidase$ Activity of Olea europaea Fruit Extracts Processed by Membrane Technology

  • Mazzei, R.;Giomo, L.;Spadafora, A.;Mazzuca, S.;Drioli, E.
    • Korean Membrane Journal
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    • v.8 no.1
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    • pp.58-66
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    • 2006
  • The ${\beta}-glucosidase$ from olive fruit is of particular interest compared to the ones from other sources because it has shown to have high specifity to convert the oleuropein into dialdehydes, which have antibacterial activity and are of high interest for their application in the food and pharmaceutical fields. The enzyme is not yet commercially available and advanced clean and safe technologies for its purification able to maintain the functional stability are foreseen. The purification of this protein from fruit extracts has been already tempted by electrophoresis but either enzyme deactivation or high background with unclear profiles occurred. In this work, fruit extracts obtained from the ripening stage that showed the highest enzyme activity have been processed by diafiltration and ultrafiltration. Asymmetric membranes made of polyamide or polysulphone having 50 and 30 kDa molecular weight cut-off, respectively, were tested for the diafiltration process. Ultrafiltration membranes made of polyethersulfone with 4 kDa molecular weight cut-off were used to concentrate the dia-filtered permeate solutions. The efficiency of the separation processes was evaluated byenzyme activity tests using the hydrolysis of p-D-nitrophenyl-${\beta}$-D-glucopyranoside (pNPGlc) as reaction model. Qualitative and quantitative electrophoresis were applied to analyze the composition of protein solution before and after the membrane separation; in addition dot blot and western blot analyses were applied to verify the presence of ${\beta}-glucosidase$ in the processed fractions. The overall results showed that the ${\beta}-glucosidase$ functional stability was preserved during the membrane operations and the removal of 20 kDa proteins allowed to increase the specific activity of the enzyme of about 52% compared to the one present in the initial fruit extract.

3D motion aftereffect in a static region after adaptation to an adjacent counterphase flickering region (역 위상 깜박임 영역 순응에 의해 유도된 인접 영역의 3차원 운동잔여 효과)

  • 김정훈;남종호;정찬섭
    • Korean Journal of Cognitive Science
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    • v.10 no.3
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    • pp.29-37
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    • 1999
  • Murakami and Cavanagh (1998a,b, 1999) reported a jitter aftereffect in a static random noise after a period of adaptation to a patch of dynamic random noise. To a account for this phenomenon. they proposed the retinal slip caused by a small eye movements in the unadapted area, which is usually compensated by the visual system to stabilize images but is unsuppressed due to the adaptation. We tested this hypothesis with new experimental method and stimuli that were supposed to nullify or reduce the effect. However. the aftereffect was still observed even under these stimuls conditions More importantly, the perceived aftereffect was rather different from Murakami and Cavanagh's. After adaptation to a counterphase flickering cosine grating, the adjacent unadapted region seems to move away from the observer during the test period instead of jittering in the frontoparallel plane. We proposed a possible explanation for this new phenomenon noting the severe contrast reduction of the adapted region during flickering period. The aftereffect might be due to the flicker-inducing contrast reduction during adaptation that produces different depth planes for the adapted and unadapted region and its restoration during the test period.

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Genetic and Phenotypic Diversity of Carbofuran-Degrading Bacteria Isolated from Agricultural Soils

  • Shin, Dong-Hyeon;Kim, Dong-Uk;Seong, Chi-Nam;Song, Hong-Gyu;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.448-456
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    • 2012
  • Thirty-seven carbofuran-degrading bacteria were isolated from agricultural soils, and their genetic and phenotypic characteristics were investigated. The isolates were able to utilize carbofuran as a sole source of carbon and energy. Analysis of the 16S rRNA gene sequence indicated that the isolates were related to members of the genera Rhodococcus, Sphingomonas, and Sphingobium, including new types of carbofuran-degrading bacteria, Bosea and Microbacterium. Among the 37 isolates, 15 different chromosomal DNA patterns were obtained by polymerase chain reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences. Five of the 15 representative isolates were able to degrade carbofuran phenol, fenoxycarb, and carbaryl, in addition to carbofuran. Ten of the 15 representative isolates had 1 to 8 plasmids. Among the 10 plasmid-containing isolates, plasmid-cured strains were obtained from 5 strains. The cured strains could not degrade carbofuran and other pesticides anymore, suggesting that the carbofuran degradative genes were on the plasmid DNAs in these strains. When analyzed with PCR amplification and dot-blot hybridization using the primers targeting for the previously reported carbofuran hydrolase gene (mcd), all of the isolates did not show any positive signals, suggesting that their carbofuran hydrolase genes had no significant sequence homology with the mcd gene.

A Study on the Effect of Optical Characteristic in 2 inch LCD-BLU by Negative and Positive Optical Pattern :I. Optical Analysis and Design (휴대폰용 2 인치 LCD-BLU의 광특성에 미치는 음각 및 양각 광학패턴의 영향 연구 :I. 광학 해석 및 설계)

  • Hwang C.J.;Ko Y.B.;Kim J.S.;Yoon K.H.
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 2006.05a
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    • pp.75-76
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    • 2006
  • LCD-BLU (Liquid Crystal Display - Back Light Unit) is one of kernel parts of LCD unit and it consists of several optical sheets(such as prism, diffuser and protector sheets), LCP (Light Guide Plate), light source (CCFL or LED) and mold frame. The LGP of LCD-BLU is usually manufactured by forming numerous dots with $50{\sim}200{\mu}m$ in diameter on it by erosion method. But the surface of the erosion dots of LGP is very rough due to the characteristics of the erosion process during the mold fabrication, so that its light loss is high along with the dispersion of light into the surface. Accordingly, there is a limit in raising the luminance of LCD-BLU. In order to overcome the limit of current dot patterned LGP, optical pattern design with $50{\mu}m$ micro-lens was applied in the present study. Especially, the negative and positive micro-lens pattern fabricated by modified LiGA with thermal reflow process was applied to the optical design of LGP. The attention was paid to the effects of different pattern conditions to the brightness distribution of BLU with micro-lens patterned LGP. Finally, negative micro-lens patterned LGP showed superior results to the one made by positive in average luminance.

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Development of ELISA for Brucella abortus RB51 II. Purification of 8kDa antigen and development of ELISA using its antigen of Brucella abortus RB51 (부루세라 RB51의 ELISA 진단법 개발 II. Brucella abortus RB51균의 8kDa 항원 정제 및 ELISA 진단법 개발)

  • Her, Moon;Cho, Dong-hee;Jung, Byeong-yeal;Cho, Seong-kun;Jung, Suk-chan;Kim, Ok-kyung
    • Korean Journal of Veterinary Research
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    • v.41 no.1
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    • pp.51-57
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    • 2001
  • A procedure for extraction and purification of 8 kDa antigen of Brucella abortus RB51 was developed. Bacteria heat inactivated at $60^{\circ}C$, 30 min was extracted by 1% sarcosine and followed by fluid pressure liquid gel filtration chromatography of 2 series, Superose 12 HR 10/30 and Sephacryl S-100. There was produced $71.46{\mu}g/g$(wet) of 8 kDa antigen, and it resisted 1% trypsin, solved 1% triton X-100 higher than distilled water and inactivated 0.1% proteinase K. These results show that 8 kDa antigen may be a lipoprotein existed cell surface of B. abortus RB51. Also, we developed ELISA using purified 8 kDa surface antigen of Brucella abortus RB51 strain, its specificity and sensitivity was 95.0%, 98.6%, respectively. As compared with dot-blot assay using whole cell and ELISA using 8 kDa antigen, its correlation was 93.5%.

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A Case Report on Pressure Ulcer Patient Treated with Fractional Laser Therapy and Korean Medicine (프락셔널 레이저 치료와 한의치료를 병행한 욕창치료 1례)

  • Kang, Ja-Yeon;Lee, Gi-Hyang;Jang, In-Soo;Kim, Hong-Jun;Jeong, Min-Jeong;Seo, Hyung-Sik
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.31 no.4
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    • pp.126-135
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    • 2018
  • Objectives : The purpose of this report to represent the efficacy of pressure ulcer treatment with fractional laser therapy and Korean medicine. Methods : We treated Parkinsonism patients with pressure ulcer using herbal medicine, fractional mode laser therapy, acupuncture and simple dressing for 3 weeks. Laser was set on fractional mode (on time $100{\mu}s$, dot pitch 0.6 mm, shape RECT) and irradiated around pressure ulcer site and center of pressure ulcer site once a day. The treatment effect was evaluated by The National Pressure Ulcer Advisory Panel(NPUAP) stage and the The Pressure Ulcer Scale for Healing(PUSH) tool(3.0). Results & Conclusions : Pressure ulcer site have improved markedly. NPUAP stage was improved from 2 to 1 and total score of the PUSH tool decreased from 12 to 8. Even though the wound was not cured completely, this shows that fractional laser therapy and Korean medicine could be effective treatment for improving pressure ulcers.

A Study on the Original Form of Slacks of Elderly Women in Their Late 60s (60대 후반 노년 여성의 슬랙스 원형 연구)

  • Moon, Soon-Ei;Park, Kil-Soon
    • The Research Journal of the Costume Culture
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    • v.19 no.5
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    • pp.929-944
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    • 2011
  • The purpose of this study was to compare and analyze two original forms of slacks for thesis to extract outward appearance of design original form applying design measures of the above through diverse research methods. The results of this study are as follow: In original form F, the original form of slacks sloper suitable to elderly women 65~9 years old was W/4+3.5+0.5cm in the front/back circumference of waist, H/4cm in the front/back circumference of hip, (H/4+1+3.8)/2cm in the position of centerline, 19cm in actual measurement value of hip length, 25cm in actual measurement value of crotch depth, (knee crotch circumference~bottom line)/2+10cm and 90 (side waist dot~ lateral malleolus length)cm in actual measurement. front dart position was 1/3 and 2/3 of front waist centerline and back dart position was 1/3 and 2/3 of back waist circumference. It was suggested that front dart quantity (length) was 3.5(11)cm, back dart quantity 3.4 (10.2)cm, side waist up quantity 0.7cm, back down quantity 1.5cm, front crotch width (H/4+1)/4-1cm, back crotch width (H/4+1)/4+4, front knee width 21.8cm, back knee width 24.4cm, front pant leg 20.8cm, and back pant leg 23.4cm.

A study on performance analysis of synchronization clock with various clock states in NG-SDH networks (NG-SDH 망에서 다양한 클럭상태 하에서의 동기클럭 성능분석에 관한 연구)

  • Lee Chang-Ki
    • The KIPS Transactions:PartC
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    • v.13C no.3 s.106
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    • pp.303-310
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    • 2006
  • This paper is to execute a study for characteristic analysis of synchronization clock and maximum network node number with various clock states, normal, SPT, LPT, in NG-SDH networks. Through the simulations, maximum network node numbers showed from 42 to 38 nodes in normal state. In SPT state, maximum network node numbers, when the last NE network applied to only SPT state, presented from 19 to 4 nodes, much less than normal state. Node numbers to meet specification in case of occurrence of SPT state in all NE networks decreased greatly. In LPT state, all maximum node numbers, when the last NE network applied to only LPT state, presented more than 50 nodes, and the results in case of occurrence of LPT state in all NE networks were also identified. However, node numbers to meet specification in case of LPT state in all DOTS networks were few large with difference between LPT and normal or SPT state.