• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.387-412
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• 2003

Influence of gelatinase on basement membrane (BM) structure was investigated by using a skin equivalent (SE) model. The results showed that (1) gelatinase produced by cells degraded the BM and (2) the addition of matrix metalloproteinase-specific inhibitor to the SE medium accelerated the formation of BM structure, indicating that gelatinase is involved in BM impairment. The activity of gelatinase was also studied in healthy human facial skin tissues. The result of in situ zymography revealed gelatinase activity around the basal layer of the epidermis, where BM integrity was severely compromised. Therefore, this enzyme was suggested to be associated with BM decomposition in human facial skin. To assess the behavior of gelatinase in stratum corneum (SC) non-invasively, an immunological study was performed. Since positive immunostaining of pro-gelatinase B was observed in SC stripped from sun-exposed skin, whereas no positive staining detected in SC of non-irradiated skin, gelatinase in the epidermis could be non-invasively detected by measuring gelatinase in SC. Gelatinase in SC of healthy female volunteers was monitored using a special film that sensitively and conveniently detects gelatinase. Ninetr percent of SC from facial skin (l00 women, 40's-50's) was gelatinase-positive. On the other hand, SC from non-irradiated skin was negative. These results strongly suggest that (1) gelatinase is constantly produced in the facial epidermis of most middle-aged woman during their daily life, and (2) the enzyme might be involved in the aging-related degeneration of both BM and the matrix fibers of the upper layer of the dermis, acting as a very important aging factor. Strong inhibitory activity against gelatinase was found in turmeric extract and identified curcumin as the major ingredient. Topical application of cream containing turmeric extract significantly decreased the number of gelatinase-positive SC clusters in human facial skins. These results indicated that turmeric is an effective ingredient to prevent skin from photo aging by suppressing chlonically upregulated gelatinase activity by UV and to improve skin condition.

• 한국발생생물학회지:발생과생식
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• 제5권1호
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• pp.23-33
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• 2001

포유류의 난자가 수란관내로 배란될 때는 난포액 성분도 같이 수란관내로 들어간다. 본 연구에서는 처음으로 난포액의 일부 성분이 수란관액에 의해서 변화하는 것을 관찰하였다. 사람의 난포액을 gelatin zymogram으로 분석한 결과 621kDa gelatinase 이외에 110kDa gelatinase (GA110) 등의 여러 gelatinase 활성이 나타났다. 이 활성들은 EDTA나 phenanthroline에 의해 억제된 반면 PMSF 처리에 의해서는 아무런 변화가 없었다. 소의 수란관액에서는 62kDa gelatinase의 활성만이 주로 관찰되었다. 소의 수란관 액과 사람의 난포액을 1:1로 섞고 이를 37$^{\circ}$C에서 3시간 동안 둔 결과 사람의 난포액의 GA110 활성은 사라졌다. 사람의 난포액에 APMA를 첨가한 결과 GA110의 활성은 대부분 감소하고 대신 62kDa gelatinase의 활성은 오히려 증가하였다. 반면에 사람의 난포액에 EDTA를 3시간 동안 처리한 결과 GA110의 활성은 오히려 현저히 증가하였고 이 때 다른 gelatinases의 활성은 영향을 받지 않았다. PMSF나SBTI는 난포액내의 gelatinases활성에 아무런 변화를 일으키지 않았다. EDTA, PMTA 혹은 SBTI 등의 proteinase inhibitor를 미리 처리한 사람의 난포액에 소의 수란관 내액을 섞은 경우에도GA110의 활성은 여전히 감소하였다. 사람의 혈청에서도 EDTA에 의해 활성이 현저히 증가하는 GA110이 발견되었다. 사람의 난포액과 유사하게 혈청내의 GA110도 소의 수란관액에 의하여 활성이 사라졌다. 그러나 사람의 난포과립세포의 추출물에서는 단지 92kDa gelatinase만 관찰이 되었다. 마지막으로 anti-human gelatinase A 항체를 사용하여 사람의 난포액과 혈청 그리고 난포과립세포의 추출액을 western blotting한 결과 621kDa과 GA110 만이 항원-항체 반응을 나타내었다. 이 같은 결과로 미루어 사람의 난포액과 혈청에는 gelatinase A의 독특한 isoform인 GA110이 있으며 특히 난포액내의 GA110은 수란관액성분에 의해 선택적으로 분해되는 것으로 여겨진다.

• 한국발생생물학회지:발생과생식
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• 제5권2호
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• pp.123-129
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• 2001

포유동물의 난포내 난자의 성숙 시에는 난자를 둘러싸고 있는 난구세포의 확장 현상이 일어나는데 이 현상에는 hyaluronic acid 뿐만 아니라 다른 성분도 관여하는 것으로 알려져 있다. 본 연구는 조직 재구성 과정에서 중요한 역할을 하는 matrix metalloproteinase(MMP)가 사람의 성숙한 난자-난구 복합체의 extracellular matrix(ECM)에 존재하는지의 여부를 알아보고자 하였다. 체외수정 시술 시에 얻어지는 사람의 난자-난구 복합체를 재료로 zymography와 western blotting 방법으로 조사한 결과 난자-난구 복합체의 ECM에는 300kDa, 240kDa, 200kDa, 180kDa, 116kDa, 97kDa, 그리고 84kDa의 분자량을 갖는 적어도 7종류의 gelatinase들이 존재하는 것이 관찰되었다. 이들 gelatinase가 MMP인지를 확인하기 위해 zymography 동안에 ethylenediaminetetraacetic acid 혹은 phenanthroline 등의 MMP 억제제를 처리한 결과 7종류 모두의 gelatinase 효소활성이 사라졌다. 또한 MMP의 활성제인 aminophenylmercuric acetate를 zymography를 시행하기 전에 ECM에 처리한 결과 200kDa, 180kDa, 97kDa, 84kDa의 gelatinase활성이 사라지고, 대신에 80kDa, 65kDa, 60kDa의 분자량을 갖는 새로운 gelatinase 단백질의 효소활성이 나타났다. 이로 미루어 사람 난자-난구 복합체의 ECM에는 여러 종류의 gelatinase들이 있으며 이들 중 일부는 MMP-2와 MMP-9의 동위효소들인 것으로 여겨진다.

• 한국발생생물학회지:발생과생식
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• 제7권2호
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• pp.105-112
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• 2003

최근 사람의 난포액에 존재하는 gelatinase 중 EDTA처리에 의해 활성이 크게 증가하는 GA110을 발견하였으며, 본 연구에서는 이러한 GA110이 만들어지는 기작을 알아보고자 하였다. 먼저, protein disulfide isomerase(PDI)가 관여하는지 알아보기 위해 PDI 저해제를 처리하여 GA110의 활성을 조사하였다. 난포액에 EDTA를 처리하기 전에 저해제를 첨가하여 반응시키면 저해제 의 농도가 증가할수록 GA110의 활성 이 감소하였으나 반면 72 kDa gelatinase의 활성은 증가하였다. 그러나 EDTA를 처 리 한 후 저해제를 첨가하여 반응시키면 GA110의 활성에 영 향을 주지 못하였다. 다음으로 72 kDa gelatinase로부터 GA110이 만들어지는 과정에 관여하는 효소를 분리하기 위하여 chromatography 방법을 이용하였으며 이렇게 분리한 효소와 기질을 반응시켜 GA110이 만들어지는 것을 확인하였다. 또한 PDI 항체를 이용하여 immunoblotting을 수행한 결과 난포액 내에 PDI보다 분자량은 약간 작지만 항체와 반응하는 단백질이 존재하는 것으로 나타났다. 이러한 결과로 보아 EDTA 처리로 인해 난포액에서 나타나는 GA110은 난포액 내에 존재하는 PDI와 유사한 효소의 활성화로 인해 72 kDa gelatinase 서로간에 이황화 결합이 형성되어 결국 72 kDa gelatinase dimer인 GA110이 만들어지는 것으로 추측된다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권2호
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• pp.407-410
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• 2015

Background: Acute kidney injury is an important issue in chemotherapy receiving patients an neutrophil gelatinase-associated lipocalin has been proposed as a novel marker. We here aimed to assess the role of urinary levels for assessment after platin exposure. Materials and Methods: Patients who had treated with cisplatin or carboplatin or oxaliplatin containg regimens were included in this study. Baseline and postchemotherapy serum urea, creatinine, urine neutrophil gelatinase-associated lipocalin and urine creatinine levels were determined. To avoid the effects of hydration during chemotherapy infusion the urinary neutrophil gelatinase-associated lipocalin/urine creatinine ratio was used to determine acute kidney injury. Results: Of a total of 42 patients receiving platin compounds,14 (33.3%) received cisplatin containing regimens, 14 (33.3%) received carboplatin and 14 (33.3%) oxaliplatin. The median age was 60 (37-76) years. Nineteen of the patients (45.2%) had lung cancer, 12 (28.6%) colorectal cancer and 11 (26.2%) others. The median pre and post chemotherapy urine neutrophil gelatinase-associated lipocalin/urine creatinin ratio was 15.6 ng/mg and 35.8 ng/mg (p=0.041) in the cisplatin group, 32.5 ng/mg and 86.3 ng/mg (p=0.004) in the carboplatin group and 40.9 ng/mg and 62.3 ng/mg (p=0.243) in the oxaliplatin group. Conclusions: Nephrotoxicity is a serious side effect of chemotherapeutic agentslike cisplatin and carbopaltin, but only to a lower extent oxaliplatin. All platin compounds must be used carefully and urine neutrophil gelatinase-associated lipocalin measurement seems to be promising in detecting acute kidney injury earlier than with creatinine.

• The Korean Journal of Physiology and Pharmacology
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• 제6권5호
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• pp.269-274
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• 2002

A large number of factors such as osteotropic hormones, cytokines, or growth factors are related to the bone remodeling which is characterized by the coupling of osteoclast-mediated bone resorption and osteoblast-mediated bone formation. Recent investigations have indicated that cytokines such as $interleukin-1{\beta}\;(IL-1{\beta})$ and tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ play a potential role in the bone resorption associated with a variety of pathological conditions such as inflammatory osteolytic disease. Collagen is the most abundant protein of the extracellular matrix of bone, and the participation of collagenase in bone resorption has been widely investigated. In this study, effects of $IL-1{\beta}$ and $TNF-{\alpha}$ on the release of collagenase from osteoblastic cells were measured. The gelatinase activity was also measured by gel substrate analysis (zymography) after electrophoresis of conditioned media of osteoblastic cell culture. $IL-1{\beta}$ increased the collagenase activity in ROS17/2.8 and HOS cell culture. $TNF-{\alpha}$ also increased the collagenase activity of osteoblastic cells. When two kinds of cytokines were treated simultaneously in the culture of osteoblastic cells, synergistic increase of collagenase activity was seen in ROS17/2.8 cells. $IL-1{\beta}$ and $TNF-{\alpha}$ significantly increased the collagenase activity after 6 hour treatment in the osteoblastic cell culture, and there was no additional increase according to the culture period. Osteoblastic cells released the gelatinase and molecular weight of this enzyme was measured about 70 KDa as assessed by zymogram. $IL-1{\beta}$ and $TNF-{\alpha}$ showed increase of the gelatinase activity produced by ROS17/2.8 and HOS cells. Taken together, this study suggested that $IL-1{\beta}$ and $TNF-{\alpha}$ can modulate bone metabolism, at least in part, by increased release of collagenase and gelatinase from osteoblasts.

• Journal of Microbiology and Biotechnology
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• 제6권6호
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• pp.445-450
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• 1996

Gelatinase A is a member of the matrix metalloproteinases that play an important role in cancer invasion and metastasis. In the course of screening gelatinase A inhibitors from microbial sources, a fungal strain PT-262 showed a strong inhibitory activity. The strain was identified as Chaunopycnis alba on the basis of its morphological characteristics. The inhibitor was isolated from acetone extract of mycelial cake by sequential chromatographies on MCI-gel, Sephadex LH-20, and a reverse-phase HPLC column. The purified inhibitor was identified as pyridoxatin by its physico-chemical properties and spectroscopic analysis. Pyridoxatin is not a peptide analog and has cyclic hydroxamic acid moiety. It inhibited activated gelatinase A with an $IC_{50}$ value of 15.2 ${\mu}M$ using fluorescent synthetic peptide. It also had a strong cytotoxicity against human cancer cell lines in vitro. Furthermore, this compound inhibited DNA synthesis with an $IC_{50}$ value of 2.92 ${\mu}M$ in PC-3 prostate cancer cells by [$^3H$]thymidine incorporation assay.

• 한국토양비료학회지
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• 제47권4호
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• pp.299-305
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• 2014

In this paper, fractional factorial screening design (FFSD) and central composition design (CCD) were used to optimize the medium components for producing chitinase and gelatinase by Lysobacter capsici YS1215. Crab shell powder, nutrient broth and gelatin were proved to have significant effects on chitinase and gelatinase activity by FFSD first. An optimal medium was obtained by using a three factor CCD, which consisted of nutrient broth of $2.0gL^{-1}$, crab shell powder of $2.0gL^{-1}$ and gelatin of $1.0gL^{-1}$, respectively with the highest chitinase activity ($3.34UmL^{-1}$) and gelatinase activity ($14.15UmL^{-1}$). This value was 3.76 and 1.11 fold of the chitinase and gelatinase activity, respectively, compared to the lowest productive medium in the design matrix. In investigating potential of these enzymes partially purified from L. capsici YS1215 for biotechnological use, the crude enzymes was found to be inhibition against pathogenic fungal mycelia: Colletotrichum gleosporioides, Phytophthora capsici, and Rhizoctonia solani. In this study, we demonstrated the optimal medium for producing the chitinolytic and gelatinolytic enzymes by the strain YS1215 and the role of their enzymes that may be useful for further development of a biotechnological use and agricultural use for biological control of phytopathogenic fungi.

• 약학회지
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• 제50권4호
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• pp.228-233
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• 2006

Matrix metalloproteinase-2 is known to be involved in pathological processes such as tumor invasion or rheumatoid arthritis. A soil microorganism producing siderophore under low iron stress $(up\;to\;5\;{\mu}m\;of\;iron)$ was identified as Burkholderia sp. Hydroxamate type siderophore produced by Burkholderia sp. CAS-5 was partially purified. MMP inhibitory activity of siderophore was confirmed by gelatin zymography. The $Zn^{2+}-chelating$ activity of siderophore correlated with the inhibition of MMP-2 activity.

• The Korean Journal of Physiology and Pharmacology
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• 제4권4호
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• pp.283-289
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• 2000

Natriuretic peptides comprise a family of three structurally related peptides; atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP). The present study was performed to investigate the effect of ANP on the proliferation and activity of ROS17/2.8 and HOS cells which are well-characterized osteoblastic cell lines. ANP dose-dependently decreased the number of ROS17/2.8 and HOS cells after 48-hour treatment. ANP generally increased the alkaline phosphatase activity of ROS17/2.8 and HOS cells after 48 hr treatment, regardless of the fact that basal activity of alkaline phosphatase was much lower in HOS cells compared to that of ROS17/1.8 cells. ANP increased the NBT reduction by ROS17/2.8 and HOS cells. ANP showed the variable but no significant effect on the nitric oxide production by ROS17/2.8 and HOS cells. ROS17/2.8 and HOS cells produced and secreted gelatinase into culture medium, and this enzyme was thought to be the gelatinase A type with the molecular weight determination. The gelatinase activity produced by ROS17/2.8 cells was increased by the treatment of ANP. However, the enzyme activity was not affected by ANP treatment in the HOS cell culture. In summary, ANP decreased the proliferation and increased the alkaline phosphatase activity and NBT reduction of osteoblasts. These results indicate that ANP is one of the important regulators of bone metabolism.