가면극과 인형극의 대비연구

김연숙 서강어문학회 1988 西江語文 Vol.6 No.1

5-플루오로우라실에 의한 사람 구강암세포주의 단백질 발현 변화에 대한 면역침전 고성능 액체크로마토그래피 분석

김연숙 대한구강악안면병리학회 2017 대한구강악안면병리학회지 Vol.41 No.6

5-fluorouracil (5-FU) is a pyrimidine analog which can work as antineoplastic antimetabolite by blocking thymidylate synthetase conversion of deoxyuridylic acid to thymidylic acid in DNA synthesis. This study is aimed to know the anticancer effect of 5-FU on the expressions of important signaling proteins in KB cells through immunoprecipitation high performance liquid chromatography (IP-HPLC). KB cells were treated with 5 µM 5-FU and cultured for 12, 24, 48, 72, and 96 hours, and followed by IP-HPLC analysis using 32 antisera. 5-FU suppressed the proliferation of KB cells by decreases in the expressions of proliferation-related proteins, Ki-67, PCNA, CDK4, and MPM2 to 82.6%, 92.4%, 95.2%, and 95.9%, respectively, but increases of antiproliferation-related proteins, p16 and p21 to 106.7% and 125.5%, respectively, during 96 hours of experiment. This proliferation reduction was also negatively regulated by cMyc/MAX/MAD network signaling. The cellular protection and survival were consistently arrested by 5-FU treatment in KB cells. The expressions of NFkB, MDR, p-mTOR, and TNFα were decreased to 95.1%, 92.8%, 93.4%, and 90.3% in 48-72 hours, respectively, while cellular stress was increased by upregulation of p38 to 111.3% in 48 hours. And the expressions of pAKT1/2/3, hTERT, and AMPK were also decreased to 93.3%, 97.4%, and 89.3% in 24-48 hours, respectively, while the cellular transformation might be undergone by upregulation of TGF-β1 to 117% until 96 hours. Particularly, 5-FU treatment greatly induced the cellular apoptosis in KB cells by increased expressions of PARP, cPARP, caspase 9, c-caspase 9, caspase 8, and caspase 3 in the lack of p53/BAX and FASL/FAS signaling. The expressions of PARP and c-PARP were increased maximum to 119.2% in 24 hours, and followed by increases of caspase 9, c-caspase 9, caspase 8, and caspase 3 to 111.2%, 125.9%, 108.6%, and 116.3% in 72-96 hours. Therefore, it is presumed that 5-FU induced cellular apoptosis in KB cells may be derived from the overexpression of PARP due to the increased DNA defect caused by 5-FU, which can lead to ATP depletion and subsequent cellular apoptosis.

Immunoprecipitation-Based High Performance Liquid Chromatography Analysis of Human Mixed Saliva

김연숙 대한구강악안면병리학회 2009 대한구강악안면병리학회지 Vol.33 No.6

As the human mixed saliva plays important roles for the protection, regeneration, immunity, and molecular transfer/signaling in the oral and gastro-intestinal mucosa, the salivary contents have great implications for the general health of human body. Nevertheless, the analysis method of human saliva has not been well developed up to date, because the proteins of mixed saliva are rapidly interacted with each other and easily degraded by proteolytic enzymes and microorganisms. This study aims to develop an immunoprecipitation-based high performance liquid chromatography (IP-HPLC) for the analysis of human mixed saliva. The representative IP-HPLC analyses were performed to compare among different subjects in variable general conditions. Compared to the normal control the subjects suffered from bacterial infections of gastro-intestinal enteritis, chronic periodontitis, and acute necrotizing gingivo-stomatitis showed dramatic increase of LL-37 level depending on the severity of diseases, while the subject suffered from Herpes stomatitis, a viral infection showed great increase of β-defensin 2. These data indicate that LL-37 in human mixed saliva is more responsible to the bacterial infections of gastro-intestinal enteritis, chronic periodontitis, and acute necrotizing gingivo-stomatitis, while β-defensin 2 is more responsible to the viral infection of Herpes stomatitis. This study also suggeststhat the IP-HPLC be easily applicable to the wide range of biological samples for the quantitative analysis of an objective protein.

면역침전 고수행 액체 크로마토그래피를 이용한 타액분석 방법의 개발

김연숙 대한구강악안면병리학회 2009 대한구강악안면병리학회지 Vol.33 No.6

As the human mixed saliva plays important roles for the protection, regeneration, immunity, and molecular transfer/ signaling in the oral and gastro-intestinal mucosa, the salivary contents have great implications for the general health of human body. Nevertheless, the analysis method of human saliva has not been well developed up to date, because the proteins of mixed saliva are rapidly interacted with each other and easily degraded by proteolytic enzymes and microorganisms. This study aims to develop an immunoprecipitation-based high performance liquid chromatography (IP-HPLC) for the analysis of human mixed saliva. The representative IP-HPLC analyses were performed to compare among different subjects in variable general conditions. Compared to the normal control the subjects suffered from bacterial infections of gastro-intestinal enteritis, chronic periodontitis, and acute necrotizing gingivo-stomatitis showed dramatic increase of LL-37 level depending on the severity of diseases, while the subject suffered from Herpes stomatitis, a viral infection showed great increase of β-defensin 2. These data indicate that LL-37 in human mixed saliva is more responsible to the bacterial infections of gastro-intestinal enteritis, chronic periodontitis, and acute necrotizing gingivo-stomatitis, while β-defensin 2 is more responsible to the viral infection of Herpes stomatitis. This study also suggeststhat the IP-HPLC be easily applicable to the wide range of biological samples for the quantitative analysis of an objective protein.

Polyacrylamide 수용액의 Dean 불안정성에 관한 연구

김연숙,김종엽 ( Yeon Sook Kim,Chong Youp Kim ) 한국화학공학회 1996 Korean Chemical Engineering Research(HWAHAK KONGHA Vol.34 No.4

Experimental studies on the Dean`s instability of aqueous solution of polyacrylamide were performed for the flow in a curved channel with the aspect ratio of 1 : 35. The range of Dean number investigated was up to 100. The critical Dean number, wave number and pressure drop vs. Dean number were examined for distilled water to validate the experimental set-up and found to be in agreement with the values reported in the literatures. The nonlinear characteristics after the onset of instability were also in good agreement with the recent report. The critical Dean number decreased with polymer concentration according to the relation Dn_c= 39 - 0.078c(Dn_c : critical Dean number; c : concentration) when c was smaller than 100 ppm. The wave number at the onset was smaller and in the range of 3-3.5. But it increased to the same value of 5.0 after the onset as in the case of Newtonian fluid. The present study reveals that both the base flow in the curved channel and Dean vortices become more unstable due to elasticity.

향요법과 복부마사지의 복합처치가 중년 여성의 체 성분 변화에 미치는 영향

김연숙,Kim, Yean-Sook 한국생활과학회 2008 한국생활과학회지 Vol.17 No.6

The purpose of this study is to determine the effect of aromatherapy, showing how the composition handling of aromatherapy and abdominal massage treatment reaches body ingredient transformation. The subjects for this research were total 18 middle-aged females in Seoul; aroma massage group of 7 females, aroma inhalation and water bathe group of 5 females, abdominal massage group of 6 females by jojoba oil without any medical effect. This clinic trial was held from July 1, 2008 to Aug.14, 2008. I held this clinic trial under the same condition after and before this clinic. A standard tape and OLYMPIA 3.5 of S hospital were used at the body measuring for subjects after and before clinic trials. I got Average and standard deviation by data analysis by SPSS Win. Ver.14.0. I did paired t-test for the comparison of before and after, and repeated measure ANOVA for between two groups or among three groups'. The verification was held Duncan Test. The results of this study were as follows: 1. Body mass quotient (F=2.86, p= .063) and Body region (F=1.34, p= .279) among three groups showed no meaningful difference, but weight meaningful difference and aroma massage group showed the greatest difference of body measure change quantity. 2. In change quantity of abdomen girth, Waist circumference and WHR, abdomen girth (F=4.56, p= .012) and Waist circumference (F=4.37, p= .031) showed a meaningful statistical difference. The result of subsequent inspection showed that there was a meaningful difference among three groups and aroma massage group was best. 3. In Cell quantity, Body region quantity and Muscle volume, Body region quantity (F=2.76, p= .182) and Muscle volume (F=3.12, p= .054) showed no difference, but Cell quantity (F=3.79, p= .040) showed a meaningful difference. In the comparison of three groups there was no difference, but aromatherapy group showed more change quantity than any other group. According to the result of this study, the composition handling of aromatherapy and body massage was effective in the decrease of Abdominal fatness and Waist circumference, Weight and the increase of Cell quantity. so I suggest that woman use this therapy in the program of obesity management for her health improvement.

‘도덕․윤리 교육의 모학문으로서 철학(윤리학)과 도덕․윤리 교과교육학의 편제’에 대한 토론

김연숙 한국윤리교육학회 2008 한국윤리교육학회 학술대회 Vol.2008 No.1

IP-HPLC Analysis of Human Salivary Protein Complexes

김연숙,이석근 대한구강악안면병리학회 2015 대한구강악안면병리학회지 Vol.39 No.5

In order to know the characteristic roles of salivary protein complex (SPC) the gel-filtration chromatography was performed using the unstimulated and the stimulated whole saliva separately. The first and second dominant SPC peaks were fractionated and analyzed by immunoprecipitation HPLC (IP-HPLC) using antibodies against the essential salivary proteins including α-amylase, mucin-1, proline rich proteins (PRPs), histatin, cystatin, LL-37, lysozyme, lactoferrin, -defensin-1, -2, -3, IgA, transglutaminase 4 (TGase 4), mucocidin, α1-antitrypsin, cathepsin G. In the gel-filtration chromatography the stimulated whole saliva showed much reduced amount of SPCs than the unstimulated whole saliva, but the proportional patterns of both whole saliva were almost similar each other. Through IP-HPLC analysis both of the first and second dominant SPCs were variably positive for the essential salivary proteins, however, α-amylase, mucin-1, PRPs, lysozyme, and cathepsin G were predominant in the first dominant SPC, while cystatin, lactoferrin, β-defensin-1, -2, -3, IgA, mucocidin, TGase 4, and α1-antitrypsin were predominant in the second dominant SPC. And more, the α1-antitrypsin and cathepsin G which were mostly derived from gingival crevicular fluid were also consistently found in the SPCs. These data may suggest that the first dominant SPC, rich in α-amylase, mucin-1, PRPs, lysozyme, and cathepsin G, may play a role in food digestion, protein degradation, and mucosa lubrication, while the second dominant SPC, rich in cystatin, lactoferrin, β-defensin-1, -2, -3, mucocidin, IgA, TGase 4, and α1-antitrypsin, may play a role in the mucosa protection and antimicrobial defense.

High Performance Liquid Chromatography Analysis of Human Salivary Protein Complexes

김연숙,이석근 대한구강악안면병리학회 2014 대한구강악안면병리학회지 Vol.38 No.6

Salivary proteins include numerous functional proteins which play important roles not only for the food-intake but also for theprotective and defensive mechanisms. In the present study the compositions of salivary proteins were analyzed by different methods,including electrophoresis and high performance liquid chromatography (HPLC). In hydrophobic protein HPLC analysis the parotid salivagradually produced macromolecular complexes when agitated in refrigerator until 30 minutes. These salivary protein complexes weredigested by neuraminidase, and then migrated more rapidly in native tris glycine gel than the control. Therefore, it was assumed thatthe glycosylated proteins of parotid saliva became gradually aggregated to form salivary protein complexes similar to those of wholesaliva. The salivary protein complexes were easily degenerated in different experimental buffers, i.e., SDS buffer, tris glycine buffer,methanol, etc., and resulted non-specific patterns in electrophoresis and HPLC. Therefore, it was presumed that the salivary proteincomplexes was made by the hydrophobic interaction as well as electrostatic attraction between salivary proteins. These data indicatedthat to know the real pattern of salivary protein complexes in vivo the whole saliva should be analyzed by HPLC using non-adheringcolumn with isoelectric buffer. Consequently, the whole saliva was analyzed by HPLC using reverse phase SuperoseTM column with20 mM potassium phosphate buffer, and two prominent peaks of salivary protein complexes were consistently found in every people. These salivary protein complex peaks were relatively stable up to 6 hours after saliva collection when the whole saliva was keptin refrigerator during experiment. Therefore, it is suggested that the salivary protein complex patterns are characteristic macromolecularstructures of whole saliva, which are also applicable as a diagnostic point in different saliva-associated diseases.

Protein Expression Changes Induced by Cisplatin in an Oral Cancer Cell Line as Determined by Immunoprecipitation-Based High Performance Liquid Chromatography

김연숙 대한구강악안면병리학회 2015 대한구강악안면병리학회지 Vol.39 No.4

Immunoprecipitation-based high performance liquid chromatography (IP-HPLC) is a type of modified enzyme-linked immunosorbent assay (ELISA) that uses protein A/G (or antibody)-conjugated beads instead of the antibody-conjugated wells used in ELISA. In order to determine the fidelity of IP-HPLC, the author used 83 antisera to identify protein expression changes caused by cisplatin treatment in KB human oral cancer cells. KB cells were cultured for 12 or 24 hours with 10 ug/mL cisplatin. The results obtained by IP-HPLC were comparable with published cisplatin data, although ELISA was not conducted in the present study. Cisplatin dominantly reduced the levels of proteins associated with cell proliferation, transcription factors, growth factors, cytoskeletal proteins, and cellular differentiating factors, but on the other hand, apoptosis-related factors, oncogenes, and protective proteins were usually up-regulated, presumably to address cisplatin-induced DNA damage. In particular, cisplatin directly inactivated genomic DNA by down-regulating histone H1 and demethylase and by up-regulating deacetylase. Cisplatin also rapidly induced p53 overexpression and mitochondria-mediated endogenous apoptosis occurred after 12 hours of cisplatin treatment, although this was almost completely replaced by FASL/FAS-mediated exogenous apoptosis after 24 hours. This preliminary study was conducted to investigate the anticancer effect of cisplatin on the KB human oral cancer cells and to determine the fidelity of IP-HPLC data. It was concluded that IP-HPLC is useful for identifying profile changes of genome wide essential proteins and signaling changes of major molecular pathways.