B3-Spline 함수를 이용한 곡선형 교량 해석

곽효경(Kwak Hyo-gyoung),최준호(Choi Joon-Ho),송종영(Song Jong-Young) 대한토목학회 2002 대한토목학회논문집 A Vol.22 No.2A

Serine pretease 억제제인 4-(2-aminoethyl) benzensulfonylfluoride (AEBSF)에 의한 호중구의 자연 세포사멸의 지연과 수지상 세포로의 전이분화 연구

박해영,곽종영,Park, Hae-Young,Kwak, Jong-Young 한국생명과학회 2007 생명과학회지 Vol.17 No.7

생체 면역반응에 중요한 역할을 하는 호중구의 세포사멸은 자연적으로 일어나거나 여러 외부자극에 의한 신호의 전달에 의해 증가하거나 지연된다. 또한 사이토카인과 같은 분화제에 의해 세포사멸이 지연되고 항원 제시 기능을 가진 수지상 세포로 분화되기도 한다. 본 연구에서는 세포사멸 억제제와 사이토카인을 이용한 시스템에서 호중구가 수지상 세포로 분화되는가를 조사하였다. Pancaspase와 serine protease의 억제제인 zVAD-fmk와 AEBSF를 처리하였을 때 호중구의 세포사멸은 현저히 감소되며 AEBSF는 caspase-3와 serine protease활성을 모두 억제하였다. 호중구의 세포사멸을 효과적으로 억제하는 AEBSF와 함께 분화제로 널리 쓰이는 CM-CSF를 같이 처리하여 3일 동안 배양하면 수지상 세포에서 높이 발현되는 CD8O, CD83 및 MHC class ll의 세포표면 마커의 발현이 증가하였다. AEBSF와 CM-CSF를 처리한 호중구를 T-세포와 함께 배양하였을 때 SEB가 존재할 경우 T-세포가 증식되었으며 SEB가 없이도 $IFN{\gamma}$는 생성되었다. 이들 결과들로 부터 serine protease 억제제인 AEBSF를 호중구에 처리하여 세포사멸을 효과적으로 억제하는 것과 수지상 세포로의 분화를 촉진하는 사이토카인인 CM-CSF의작용을 나타내게 하는 조건과는 서로 상호적으로 연관되어 작용할 수 있다는 것을 제시하고 있다. Neutrophils play a key role as a first line of defense and are known to acquire the characteristics of dendritic cells (DCs) under the appropriate conditions. The spontaneous apoptosis of neutrophils was delayed by treatment with 4-(2-aminoethyl) benzensulfonylfluoride (AEBSF), a serine protease inhibitor. AEBSF inhibited both caspase-3 and serine protease activities, whereas ZVAD-fmk, a pancaspase inhibitor, inhibited only caspase-3 activity. The life span of neutrophils was prolonged up to 5 days by AEBSF in the presence or absence of granulocyte macrophage colony stimulating factor(CM-CSF). DC surface markers, such as CD80, CD83, and MHC class ll were not expressed on neutrophils treated with AEBSF alone. CM-CSF failed to prolong the survival time of neutrophils up to3 days but increased the expression levels of DC markers on neutrophils in the presence of AEBSF. Expression levels of DC markers were the highest on neutrophils treated with CM-CSF and AEBSF for 3 days. AEBSF and CM-CSF-treated neutrophils stimulated proliferation of T cells in the presence of a superantigen, Staphylococcal enterotoxin B (SEB) but produced $interferon-{\gamma}$ ($IFN{\gamma}$) in the absence of SEB. These results suggest that the inhibition of serine protease activity prolonged the life span of human neutrophils and combined treatment of neukophils with CM-CSF and serine protease inhibitor induced differentiation of neutrophils into DC-like cells.

ACE 삽입 및 결손 유전자 다형성과 HDL 콜레스테롤과의 관련성

유창훈,곽종영,김나영,노미숙,정갑열,이용환,김정만,김준연,홍영습,You, Chang-Hun,Kwak, Jong-Young,Kim, Na-Young,Roh, Mee-Sook,Jung, Kap-Yeol,Lee, Yong-Hwan,Kim, Jung-Man,Kim, Joon-Youn,Hong, Young-Seoub 대한예방의학회 2006 예방의학회지 Vol.39 No.6

Objectives : The purpose of this study is to evaluate the association of the angiotensin converting enzyme (ACE) insertion/deletion (I/D) polymorphism with cardiovascular disease risk factors. Methods : Out of a total of 608 middle-aged adults who visited local health centers, 424 subjects (104 male, 320 female) who had not been diagnosed with hypertension, diabetes mellitus, or hyperlipidemia were included in this study. ACE genotypes were determined in all subjects by polymerase chain reaction methods. Results : Statistical differences in high-density lipoprotein (HDL) cholesterol levels according to ACE genotype were observed using ANOVA (p<0.05), but no differences were found in other cardiovascular risk factors. Specifically, men with the DD and DI genotypes had significantly lower HDL cholesterol levels than those with the 11 genotype based on the LSD multi-comparison test (p<0.05). Conclusions : In men, the D-allele of the ACE I/D polymorphism was significantly associated with reduced HDL cholesterol levels. In the future, larger studies are needed to confirm this relationship between ACE I/D polymorphism and HDL cholesterol.

한국인에서 peroxisome proliferator-activated receptor alpha Leu162Val 유전자 다형성과 대사증후군간의 관련성

신승철,송혜순,홍영습,곽종영,유병철,이용환,Shin Soung-Cheal,Song Hye-Soon,Hong Young-Seoub,Kwak Jong-Young,Yoo Byung-Chul,Lee Yong-Hwan 한국생명과학회 2006 생명과학회지 Vol.16 No.2

Peroxisome proliferator-activated receptors alpha (PPAR $\alpha$)는 지질대사와 관련하여 대사증후군 발생과 관련이 있을 수 있는 강력한 잠재 유전자로 고려되고 있으므로 한국인에 있어서 PPAR$\alpha$ L162V 유전자 다형성과 대사증후군과의 연관성을 확인하고자 고신대학교 복음병원에서 2004년 12윌에서 2005년 7월 사이에 건강진단을 받았던 수진자 542명(대사 증후군 : 262명, 정상인 : 280명)을 대상으로 신장, 체증, 체질량지수, 허리둘레와 수축기와 이완기 혈압, 공복 혈당, 총콜레스테롤, HDL 콜레스테롤, LDL 콜레스테롤과 중성지방 수치를 측정하였으며, 대사증후군의 정의는 혈압, 공복 혈당, HDL 콜레스테롤, 중성지방은 NCEP ATP III의 기준을 적용하였고, 허리둘레는 WHO 아시아-서태평양 기준을 적용하였다. PCR-ASO (polymerase chain reaction allele-specific oligonucleotide) 방법에 의해 대상자들의 PPAR$\alpha$ L162V 유전자 다형성을 확인하였다. 연구결과 PPAR$\alpha$ 484번 염기서열의 $C{\rightarrow}G$ 돌연변이가 나타난 사람은 조사대상자 542명 가운데 1명(0.2%) 이었다. 한국인에서는 PPAR$\alpha$ L162V 유전자 다형성이 거의 일어나지 않았으며, 이의 확인을 위하여 더욱 많은 사람을 대상으로 연구가 진행되어야 할 필요가 있을 것으로 생각된다. Peroxisome proliferator activated receptor (PPAR)-$\alpha$ of three PPAR subtypes ($-\alpha,\;-\beta/-\gamma,\;-\delta$), which are members of the nuclear hormone receptor superfamily of ligand-activated transcription factors, plays a key role in lipoprotein and glucose homeostasis. A variation in the PPAR-a gene expression has been suggested to influence the development of metabolic syndrome through alterations in lipid concentrations. The aim of our study was to investigate the association between the PPAR-a and metabolic syndrome among South Korean. A total of 542 health screen examinees were enrolled in this study who were examined in Kosin University Gospel Hospital from December, 2004 to July, 2005. The height, weight, waist circumference, and systolic and diastolic blood pressure of the subjects were examined and fasting blood glucose, total cholesterol, HDL cholesterol, LDL cholesterol, triglyceride were measured by-sampling in venous blood. The metabolic syndrome was defined as the presence of three or more of the following : waist circumference men ${\geq}90cm$, women ${\geq}80cm$, blood pressure ${\geq}130/85mmHg$, fasting glucose ${\geq}110mg/dL$, HDL cholesterol men <40 mg/dL, women <50 mg/dL, triglyceride ${\geq}150mg/dL$. The blood pressure, fasting glucose, HDL cholesterol, triglyceride were evaluated by using the criteria of NECP ATP III and waist circumference was assessed by using the criteria of WHO Asia-Western Pacific. And the author compared the frequency of the PPAR-$\alpha$ mutation of L162V ($C{\rightarrow}G$ variant in exon 5) in a sample of 542 subjects with and without the metabolic syndrome by polymerase chain reaction allele-specific oligonucleotide (PCR-ASO) method. One (0.2%) hetero-isotype among high risk of metabolic syndrome was identified. The values of waist circumference, body mass index and low density lipoprotein cholesterol of the mutant were 100 cm, 28.6 $kg/m^2$ and 120 mg/dL, respectively. Although the author failed to see significant association between the presence of the PPAR-$\alpha$ L162V polymorphism and metabolic syndrome, one PPAR-$\alpha$ L162V polymorphism in metabolic syndrome patients was found.

양수내 세포 및 양막세포주 WISH 세포에서 아스피린에 의한 prostaglandin H synthase의 발현 증가

차문석(Moon Seok Cha),곽종영(Jong Young Kwak) 대한산부인과학회 1998 Obstetrics & Gynecology Science Vol.41 No.11

N/A Prostaglandins which are produced from amnionic cells are known to play a major role in uterine contraction and cervical dilatation in human. Recently it is reported that aspirin increases the expression of PGHS-1 and PGHS-2 in trophoblast cells from placenta. We examined here the changes of immunoreactive prostaglandin H synthase (PGHS) level by aspirin in cultured cells from amniotic fluid and human amnionic WISH cells. PMA (10-7 M), an activator of protein kinase C increased the induction of PGHS-2 in both cells with or without fetal calf serum. PGHS-2 protein was also increased significantly by 10-4 M aspirin at 6 hours in both cells in the presence of serum but it was not increased in the absence of serum. The expression of PGHS-1 protein was enhanced by asprin but not by PMA in the absence of serum. Other anti-inflammatory drugs such as acetaminophen, indomethacin, dexame- thasone, and mefenamic acid increased the PGHS-2 protein level in WISH cells. PMA-induced PGHS-2 expression in WISH cells was not decreased by aspirin, on the contrary, the level was increased additively. Our results show that the increased expression of PGHS in amnion cells or other amniotic fluid cells by aspirin and other several anti-inflammatory drugs is through an unidentified effect rather than feedback effect by depletion of prostaglandin.

변압기 절연유 가스분석과 고장원인 검토

權東震(Dong-Jin Kweon),郭周植(Joo-Sik Kwak),殷鍾榮(Jong-Young Eun),閔丙文(Byeong-Moon Min),庾東均(Dong-Gyon Yu) 대한전기학회 2005 전기학회논문지C Vol.54 No.8

혈관 내피세포 성장 인자에 의한 호중구 자연 세포사멸의 지연효과

전창원(Chang-Won Jeon),박해영(Hae-Young Park),곽종영(Jong-Young Kwak),김형호(Hyung-Ho Kim) 대한외과학회 2005 Annals of Surgical Treatment and Research(ASRT) Vol.69 No.4

프레닐 페놀계 항생제인 4-O-methyl-ascochlorin에 의한 호중구 세포사멸의 유도

손동훈,이선영,이민정,박주인,홍영습,이용환,장영채,곽종영,Son Dong-Aoon,Lee Sun-Young,Lee Min-Jung,Park Joo-In,Hong Young-Seob,Lee Yong-Hwan,Chang Young-Chae,Kwak Jong-Young 한국생명과학회 2006 생명과학회지 Vol.16 No.1

호중구의 세포사멸은 자연적으로 일어나지만 여러 외부자극에 의한 신호의 전달에 의하여 증가하거나 지연된다. 본 연구에서는 항암, 항생제로 개발된 프레닐 페놀계인 ascochlorin의 유도체 중에서 백혈구 암의 세포사멸을 유도하는 4-O-methyl-ascochlorin (MAC)이 호중구의 자연 세포사멸 및 지연되는 세포사멸에 어떠한 영향을 미치는가와 그 작용기작을 연구하였다. 호중구의 세포사멸은 사람 말초 혈액으로부터 분리하여 세포 배양 시간에 따라 형태 변화, annexin-V/propidium iodide의 염색, 및 DNA 전기영동 등으로 조사하였다. MAC는 농도 및 시간 의존 형으로 호중구의 세포사멸을 증가시켰다. 그러나 granulocyte macrophage-colony stimulating factor나 lipopolysaccharide 등에 의한 세포사멸의 지연은 MAC에 의하여 부분적으로 억제되었다. MAC에 의한 세포사멸의 유도는 pancaspase, caspase-8 및 caspase-3 억제제인 zVAD-fmk. zIETD-fmk, 및 zDEVD-fmk에 의하여 억제되었으며 procaspase-8과 procaspase-3의 단백질 양도 MAC로 처리한 호중구에서 현저히 감소하였다. 미토콘드리아 막 투과성은 MAC에 의하여 현저히 감소하였으나 zVAD-fmk에 의하여 완전히 봉쇄되지 못하였다. 이들 결과 들은 MAC에 의한 호중구 세포사멸의 증가는 caspase-8 및 caspase-3의 활성을 통하여 일어나지만 미토콘드리아의 막성분에는 영향이 없다는 것을 제시하고 있다. Neutrophils are short-lived leukocytes that play a vital role in immune responses to bacteria, yeast, and fungi. This study was performed to investigate the effect of 4-O-methyl-ascochlorin (MAC), an anti-tumor, antibiotic, and anti-fungal prenyl-phenol compound on the spontaneous apoptosis of human neutrophils. MAC time- and dose-dependently accelerated the spontaneous apoptosis of human neutrophils. The effect of MAC on neutrophil apoptosis was blocked by pre-treatment of the neutrophils with specific inhibitors of pancaspase (zVAD-fmk), caspase-8 (zIETD-fmk), or caspase-3 (zDEVD-fmk). The cleavage of procaspase-8 and procaspase-3 was increased by MAC. Mitochondrial permeability, which was measured by the retention of $DiOC_6(3)$, was dose-de-pendently increased by MAC but the change of mitochondrial permeability was not blocked by pretreatment of neutrophils with zIETD-fmk. These results suggest that MAC induces neutrophil apoptosis by caspase-8-dependent but mitochondria-independent manner.

소음성 난청에서의 Mitochondrial DNA A3243G, A1555G, A7445G 돌연변이

홍영습,이명진,곽기영,황찬호,신동훈,곽종영,이용환,김종민,김준연,Hong Young-Seoub,Nishio Hisahide,Lee Myeong-Jin,Kwak Ki-Young,Hwang Chan-Ho,Shin Dong-Hoon,Kwak Jong-Young,Lee Yong-Hwan,Kim Jong-Min,Kim Joon-Youn 한국생명과학회 2004 생명과학회지 Vol.14 No.6

본 연구는 소음성 감각신경성난청 환자의 유전적 관련요인을 파악하고자 관련성이 의심되는 mitochondrial DNA의 돌연변이와 소음성 감각신경성난청과의 관련성을 조사하였다. 말초혈액 백혈구로부터 DNA를 추출한 후, mtDNA 3243, 1555, 7445부위의 $A{\rightarrow}G$ 돌연변이 유무를 관찰하기 위하여 mtDNA 3243, 1555, 7445부위 가 포함된 mtDNA fragment를 중합효소 연쇄반응으로 증폭하고 유전자 제한효소로 소화하여 전기영동하고 ethidium bromide 용액으로 염색하여 UV transilluminator에서 관찰하였다. 그리고, PCR 산물을 이용하여 DNA 염기서열을 분석하여 mtDNA 3243, 1555, 7445부 위에서의 염기서열 분석을 실시하여 mtDNA 3243, 1555, 7445부위 의 $A{\rightarrow}G$ 돌연변이를 관찰하였다 MtDNA A3243G, A1555G, A7445G의 돌연변이를 관찰한 결과 돌연변이 부위가 포함된 fragment가 소음성 감각신경성난청 환자군, 감각신경성난청 환자군, 대조군 모두에서 증폭됨을 관찰하였다. 또한 PCR 산물을 제한효소로 처 리 한 결과에서도 mtDNA에서 3243, 1555, 7445부위의 $A{\rightarrow}G$ 돌연변이가 일어나지 않았음을 알 수 있었다. PCR산물을 이용하여 DNA 염기서열을 분석하여 mtDNA 3243, 1555, 7445부위에서의 염기서열을 확인한 결과 이미 밝혀진 사람의 mtDNA 3243, 1555, 7445부 위의 염기서열과 동일한 염기서열임이 확인되었으므로 mtDNA 3243, 1555, 7445부위의 $A{\rightarrow}G$ 돌연변이가 일어나지 않았음을 확인하였다. 소음성 감각신경성난청과 mtDNA 3243, 1555, 7445부위의 $A{\rightarrow}G$ 돌연변이와는 관련이 없는 것으로 관찰되었다. Mitochondrial DNA mutations have been reported in recent years in association with sensorineural hering loss. The purpose of this study is to identify the association between the noise-induced sensorineural hearing loss and the A to G mutation at nucleotide 3243, 1555, 7445 of mitochondrial DNA. Study subjects were established by history and chart review, and audiological and clinical data were obtained. Blood was sampled from 214 normal controls, 102 noise-induced hearing loss, and 28 sensorineural hearing loss. The DNA of these individuals were extracted, and mitochondrial DNA fragments were analyzed by polymerase chain reaction. Subsequently, the coding sequence of mitochondrial DNA 3243, 1555, 7445 were sequenced, and compared to the normal sequence, and all sequence variations were analyzed by restriction enzymes. Mitochondrial DNA mutations $(3243A{\rightarrow}G,\;1555A{\rightarrow}4G,\;7445A{\rightarrow}G)$ were not detected by polymerase chain reactions in any patients with noise-induced hearing loss, sensorineural hearing loss, and normal controls. The DNA sequencing of PCR products did not revealed an A to G substitution at nucleotide 3243, 1555, 7445 of mitochondrial DNA. The noise-induced sensorineural hearing loss was not associated with mitochondrial DNA mutation $(3243A{\rightarrow}G,\;1555A{\rightarrow}4G,\;7445A{\rightarrow}G)$.

중증 자궁내막증 환자의 복강액이 단핵구의 HLA-DR 발현에 미치는 영향

나용진 ( Yong Jin Na ),박종훈 ( Jong Hoon Park ),곽종영 ( Jong Young Kwak ),이규섭 ( Kyu Sup Lee ) 대한산부인과학회 2006 Obstetrics & Gynecology Science Vol.49 No.12

목적: 자궁내막증은 자궁강 밖에 자궁내막 조직이 존재하는 질환으로 가임기 여성에서 흔한 질환이다. 발생기전으로는 월경혈의 역류, 자궁내막 세포의 이소성 착상, 유전학적 인자, 그리고 호르몬 이상 등이 제시되고 있으나 아직 명확하지 않다. 최근에는 자궁내막 세포의 이소성 착상 기전을 면역학적 내성과 관련하여 설명하고 있는 등, 자궁내막증의 발생에서 면역학적 기전의 중요성이 점차 강조되고 있다. 본 연구는 자궁내막증의 발생과 관련된 면역학적 인자에 관한 분석을 위하여 자궁내막증 환자의 복강액이 단핵구에서의 HLA-DR 발현에 미치는 영향을 알아보고자 하였다. 연구 방법: 가임기 여성으로 복강경 검사를 받은 환자들 중 자궁내막증 환자의 복강액 (n=20)과 대조군의 복강액 (n=10)을 본 수술 시작 전에 캐뉼러를 이용하여 채취하였다. 건강한 공여자로부터 얻은 말초혈액 단핵구는 분리배지 (Histopaque- 1077)를 이용하여 분리, 배양하였고, THP-1 세포는 RPMI 1640 배지를 이용하여 배양하였다. 단핵구에서의 HLA-DR의 발현은 동일초점 현미경으로 관찰하였고, IL-10, TGF-β와 VEGF의 농도는 효소결합면역흡착검사를 이용하여 측정하였다. 결과: 자궁내막증 환자의 복강액은 단핵구에서의 HLA-DR 발현을 감소시켰으며 (P<0.01), 첨가된 자궁내막증 환자의 복강액 농도가 증가할수록 THP-1 세포의 HLA-DR 발현이 용량 의존성으로 감소하였다. 또한 자궁내막증 환자의 복강액 내 IL-10 농도와 단핵구의 HLA-DR 발현은 역상관관계를 보였다 (r=-0.518, P=0.019). 결론: 단핵구의 HLA-DR 발현은 자궁내막증 환자의 복강액에 의해 IL-10과 연관되어 억제되며, 이는 자궁내막증 환자의 복강액 내 IL-10이 단핵구의 HLA-DR의 발현을 조절하는 중요한 요소(들) 중 하나라는 것을 의미한다. Objective: Endometriosis is a common gynecologic disorder characterized by the presence of endometrial tissue outside the uterine cavity. Many theories have been suggested to explain the mechanism for the development of this disease. Although no single theory can explain all cases of endometriosis, retrograde menstruation theory has gained the widest acceptance. But the discrepancy between the incidence of retrograde menstruation and prevalence of endometriosis suggests that additional factor(s) may determine susceptibility to endometriosis in the peritoneal cavity and the survival and implantation of endometrial cells seem to be related with immunologic factors. For evaluation of the immunologic factors, we investigated the effects of peritoneal fluid from patients with advanced endometriosis (EPF) on the HLA- DR expression of monocytes. Methods: EPF (n=20) and peritoneal fluid from the control group (CPF) (n=10) were collected prior to main laparoscopic operation. CPF were obtained from fertile women with various gynecologic problem other than endometriosis, such as dermoid cyst and carcinoma in situ of the uterine cervix. Peripheral blood mononuclear cells were collected by density-gradient centrifugation of whole blood over a peripheral blood mononuclear cell separation medium (Histopaque-1077), and THP-1 cells derived from a monocyte/macrophage cell line (Korean Cell Line Bank, Seoul, Korea) were cultured in RPMI-1640 medium. The expression of HLA-DR was evaluated by confocal microscopy, and the levels of IL-10, TGF-β and VEGF were measured by standard enzyme-linked immunosorbent assay kits (R and D system). Results: Compared to CPF, the addition of 10% EPF (n=10) to culture medium significantly reduced the percentage of HLA-DR positive cells in cultures of a THP-1, monocytic cell line at 48 hours. The effect of EPF was dose-dependent, and similar effect was observed in the peripheral blood monocytes. An inverse correlation was found between the HLA-DR expression level and IL-10 concentration in EPF (r=-0.518; P=0.019) and in the supernatant of peripheral blood monocyte cultured in EPF (n=20) (r=-0.459; P=0.042). IL-neutralizing antibody significantly abrogated the effect of EPF on HLA-DR expression level. Conclusion: These results suggest that HLA-DR expression level on monocytes is down-regulated by EPF, and that IL-10 may be one of the factors in EPF to modulate HLA-DR expression level.