• 제목/요약/키워드: gelatin type

검색결과 74건 처리시간 0.021초

Purification and Cloning of an Extracellular Serine Protease from the Nematode-Trapping Fungus Monacrosporium cystosporium

  • Yang, Jin-Kui;Ye, Feng-Ping;Mi, Qi-Li;Tang, Song-Qing;Li, Juan;Zhang, Ke-Qin
    • Journal of Microbiology and Biotechnology
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    • 제18권5호
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    • pp.852-858
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    • 2008
  • An extracellular protease (Mc1) was isolated from the nematode-trapping fungus Monacrosporium cystosporium by gel filtration, anion-exchange, and hydrophobic interaction chromatographies. This protease had a molecular mass of approximately 38 kDa and displayed an optimal activity at pH 7-9 and $56^{\circ}C$ (over 30 min). Its proteolytic activity was highly sensitive to the serine protease inhibitor PMSF (phenylmethylsulfonylfluoride, 0.1 mM), indicating that it belonged to the serine-type peptidase group. The Michaelis constant ($K_m$) and $V_max$ for substrate N-Suc-Ala-Ala-Pro-Phe-pNA were $1.67{\times}10^{-4}\;M$ and 0.6071 $OD_{410}$ per 30 s, respectively. This protease could degrade a broad range of substrates including casein, gelatin, BSA (bovine serum albumin), and nematode cuticle. Moreover, the enzyme could immobilize the free-living nematode Panagrellus redivivus and the pine wood nematode Bursaphelenchus xylophilus, suggesting that it might playa role in infection against nematodes. The encoding gene of Mc1 was composed of one intron and two exons, coding for a polypeptide of 405 amino acid residues. The deduced amino acid sequence of Mcl showed 61.4-91.9% identity to serine proteases from other nematode-trapping fungi. Our results identified that Mcl possessed biochemical properties including optimal reaction condition and substrate preference that are different from previously identified serine proteases.

Matrix metalloproteinases: expression and regulation in the endometrium during the estrous cycle and at the maternal-conceptus interface during pregnancy in pigs

  • Inkyu Yoo;Soohyung Lee;Yugyeong Cheon;Hakhyun Ka
    • Animal Bioscience
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    • 제36권8호
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    • pp.1167-1179
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    • 2023
  • Objective: Matrix metalloproteinases (MMPs) are a family of endoproteases produced by various tissues and cells and play important roles in angiogenesis, tissue repair, immune response, and endometrial remodeling. However, the expression and function of MMPs in the pig endometrium during the estrous cycle and pregnancy have not been fully elucidated. Thus, we determined the expression, localization, and regulation of MMP2, MMP8, MMP9, MMP12, and MMP13 in the endometrium throughout the estrous cycle and at the maternal-conceptus interface during pregnancy in pigs. Methods: Endometrial tissues during the estrous cycle and pregnancy and conceptus and chorioallantoic tissues during pregnancy were obtained and the expression of MMPs was analyzed. The effects of steroid hormones and cytokines on the expression of MMPs were determined in endometrial explant cultures. Results: Expression levels of MMP12 and MMP13 changed during the estrous cycle, while expression of MMP2, MMP9, MMP12, and MMP13 changed during pregnancy. Expression of MMP2, MMP8, and MMP13 mRNAs was cell type-specific at the maternal-conceptus interface. Gelatin zymography showed that enzymatically active MMP2 was present in endometrial tissues. In endometrial explant cultures, estradiol-17β induced the expression of MMP8 and MMP12, progesterone decreased the expression of MMP12, interleukin-1β increased the expression of MMP2, MMP8, MMP9, and MMP13, and interferon-γ increased the expression of MMP2. Conclusion: These results suggest that MMPs expressed in response to steroids and cytokines play an important role in the establishment and maintenance of pregnancy by regulating endometrial remodeling and processing bioactive molecules in pigs.

등골나물 추출물이 인간의 유방암세포인 MDA-MB-231 세포의 이동, 침윤 및 부착에 미치는 영향 (Effect of Eupatorium japonicum Extract on the Metastasis, Invasion and Adhesion of MDA-MB-231 Human Breast Cancer Cells)

  • 우은영;박소영;권수진;권규택;김종대;임순성;윤정한
    • 한국식품과학회지
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    • 제43권2호
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    • pp.213-219
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    • 2011
  • 등골나물은 국화과 여러해살이 식물로 한방에서는 고혈압, 폐렴, 황달, 홍역, 요통 등에 사용한다고 알려져 있다. 본 연구에서는 등골나물의 꽃 부위를 추출하여 등골나물 추출물이 유방암 세포인 MDA-MB-231 세포의 이동, 침윤 및 부착에 미치는 영향을 조사하였다. 그 결과 MDA-MB-231 세포의 이동, 침윤 및 부착은 등골나물 추출물의 농도($0-20{\mu}g/mL$)가 증가할수록 현저하게 감소하였다. 등골나물 추출물은 MMP-9, MMP-2의 활성을 억제하였고, TIMP-1의 발현은 감소시킨 반면 TIMP-2의 발현은 증가시켰다. 또한, 등골나물 추출물은 uPA, VEGF 그리고 ICAM의 mRNA 및 단백질 수준을 현저히 감소시켰다. 특히, 등골나물 헥산 분획물이 유방암세포의 이동을 현저하게 억제하였다. 이상의 결과로부터 등골나물 추출물은 MMP-9, MMP-2, uPA, TIMP-1 및 ICAM의 감소, TIPM-2의 증가를 통해 유방암세포의 전이를 억제하는 것으로 판단된다. 따라서 본 연구는 이러한 효능을 지닌 등골나물 추출물을 암전이에 효과가 있는 암예방제나 항암제로 개발할 수 있는 가능성을 제시한다.

패류중의 저온성 대장균군 및 세균 Flora에 관한 연구 (A STUDY ON PSYCHROTROPHIC COLIFORM AND BACTERIAL FLORA IN SHELLFISH)

  • 신석우;강성구
    • 한국수산과학회지
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    • 제12권1호
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    • pp.19-26
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    • 1979
  • 패류에서 저온성 대장균군의 위생학적 의의 및 패류의 위상상태를 평가하기 위한 자료를 얻고자 1978년 1월부터 12월까지 매월 참굴, 바지락, 홍합, 고막을 시료로하여 실험한 결과를 요약하면 다음과 같다. 1. 균수의 범위는 생균수가 $1.3\times10^3-2.5\times10^6/g$, 대장균군이 $>\~5.5\times10^2/g$, E. coli MPN이 $2\~9$. $2\times10^2/100g$으로 생균수는 4월에서 10월사이에는 대부분의 시료에서 패류의 위생학적 기준치를 초과하였고 E. coli 최확수도 7검체에서 초과한 것으로 나타났다. 2. 대장균군 373균주중 298균주가 분류동정되었고 이 가운데서 E. coli 1이 138균주$(46\%)$, K. aerogenes 1이 71균주$(46\%)$였다. 3. $5^{\circ}C$에서 발육가능한 저온성 대장균군은 분리균 373균주중에서 186균주로 $50\%$였다. 4. 세균 flora는 분리균 453균주 가운데서 Vibrio가 240균주$(53\%)$, Pseudomonas가 91균주$(20\%)$로 이들 2균속이 우세한 세균류를 구성하고 있었다. 5. 세균류의 계절적 변동은 Vibrio가 5월에서 10월사이 분포도가 높은 경향을 나타냈고 Moraxella 동계아만 검출되었으며 그 외의 균속은 큰 변동이 보이지 않았다. (본 연구는 1977년도 문교부 학술연구 조성비에 의한 것임).

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치은섬유아세포의 MMP 발현에 대한 Nitric Oxide의 영향 (Nitric Oxide on the MMP-2 expression by human gingival fibroblasts)

  • 신인식;윤상오;정현주;고정태
    • Journal of Periodontal and Implant Science
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    • 제33권2호
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    • pp.277-288
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    • 2003
  • It has been suggested that increased number and activity of phagocytes in periodontitis lesion results in a high degree of reactive oxygen species (ROS) such as superoxide anion, hydrogen peroxide, nitric oxide and peroxynitrite. There are few reports on the relationship between ROS and MMPs expressions in gingival fibroblast. We studied to elucidate whether and how ROS, especially nitric oxide affects the MMP expression. Human gingival fibroblasts and HTl080 cells (human fibrosarcoma sell line as reference) were grown in DMEM supplemented with 10 mM HEPES, 50 mg/L gentamicin, and 10% heat inactivated fetal bovine serum with addition of various reactive oxygen species (ROS). Culture media conditioned by cells were examined by gelatin zymography. HT1080 cells expressed proMMP-2 and proMMP-9, but human gingival fibroblasts (HGF) produced only proMMP-2. Hydrogen peroxide upregulated MMP-9 expression in HT1080 cells, whereas in human gingival fibroblast SNP treatment showed marked increase in MMP-2 level compared to other ROS. These results suggest that the effects of ROS on MMPs expressions are cell-type specific. RT-PCR for MMP-2 and TIMP-2 m-RNA were performed using total RNA from cultured cells under the influence various kinase inhibitors. In HT1080 cells, treatment with FPTI III (Ras processing inhibitor) and LY294002 (PI3-kinase inhibitor) resulted in inhibition of MMP-2 and MMP-9 expressions, suggesting that Ras/P13-kinase pathway is important for MMPs expression in HT1080 cells. In gingival fibroblasts, treatment with FPTI III and PDTC (NF-kB inhibitor) showed marked decrease in MMP-2 regardless of the of SNP , suggesting that Ras/NF-kB could be the key pathway for NO-induced MMP-2 expression in gingival fibroblasts. This study showed that ROS, especially nitric oxide, could be the critical mediator of periodontal disease progression through control of MMP-2 expression in gingival fibroblasts possibly via Ras/NF-kB pathway.

구리의 선택적 전착에서 결정 입자의 크기가 전기적 접촉성에 미치는 영향 (Effect of the particle size on the electrical contact in selective electro-deposition of copper)

  • 황규호;이경일;주승기;강탁
    • 한국결정성장학회지
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    • 제1권2호
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    • pp.79-93
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    • 1991
  • 초 고집적 회로의 시대로 접어들면서 지금까지의 금속선 형성 기술 및 배선 재료에 많은 문제점들이 나타나고 있다. 알루미늄의 대체 재료로서 검토되고 있는 구리를, 전기 화학적 방법에 의해 미세 접촉창에 선택적으로 충전함으로써 새로운 금속선 형성 기술을 제시하고자 하였다. 0.75M의 황산구리 수용액을 전해액으로 사용하여 p형 (100) 규소 박판위에 구리 전착막을 형성한 후 Alpha Step, 주사 전자 현미경, 4-탐침법을 사용하여 막의 두께, 입자 크기, 비저항을 측정함으로써 전착 시간, 전류 밀도, 첨가물로 사용한 젤라틴 농도가 전착막의 성질에 미치는 영향에 대해 조사하였다. 평균 전착 속도는 전류 밀도가 $ 2A/dm^2$일 때 0.5-0.6\mu\textrm{m}$/min 였고 구리 입자의 크기는 전류밀도 증가에 따라 증가하였다. 입자 크기 $4000{\AA}$이상에서 얻어진 비저항값은 3-6 Ω.cm였다. 젤라틴을 첨가하여 입자의 크기를 $0.1\mu\textrm{m}$이하로 감소시킴으로써 크기 $1\mu\textrm{m}$이하의 접촉장에 구리를 선택적으로 충전시키는데 성공하였다.

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국내 연근해 및 환자로부터 분리된 vibrio vulnificus의 세균학적 특징

  • 신광훈;신영학;이종삼
    • 미생물학회지
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    • 제30권1호
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    • pp.15-29
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    • 1992
  • 국내 연안의 어패류, 해구 및 환자 등으로부터 Vibrio vulnificus를 분리동정하여 생화학적 성상, 생장조건, 혈천형, 약제 감수성 시험 등 세균학적 특성을 밝혔다. V. vulnificus 균의 생화학적 성상은 유당을 발효(95%)하고 서당은 발효 (4%)않은 균주가 대부분이었으며, ornithine 발효능, gelatine 액화능, mannitol 분해능 등이 다른 연구자들과 동일한 결롸를 나타내지 않았다. 따라서 이들 성상이 V. vulnivicus 동정에 그리 유용하지 않다. o 혈청형은 자연계 분리균주의 경우 o2 혈청형이 제일 많았고 (24%) 환자 분리균주의 경우 04 균(42%)이 제일 많았다. V. vulnificus 의 최적 생상조건은 염농도는 306%였고, 온도는 25-35.deg.C, pH는 8로 나타났다. V. vulnificus균은 chloramphenicol, erythromycin, tetracycline, ampicilin 에 높은 감수성을 지니고 있었고 gentamycin 에 대하여는 내성을 가진 것으로 나타났으며, 자연계 분리균주가 환자 분리균주보다 이들 항생제에 더 높은 감수성을 지니고 있었다. chelating agent EDTA가 V. vulnificus에 대하여 생장억제효과가 있음이 밝혀졌으나 치사적으로 작용치는 않았다.

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Microbial Exposure Assessment in Sawmill, Livestock Feed Industry, and Metal Working Fluids Handling Industry

  • Park, Hyun-Hee;Park, Hae-Dong;Lee, In-Seop
    • Safety and Health at Work
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    • 제1권2호
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    • pp.183-191
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    • 2010
  • Objectives: The objective of this study is to investigate the distribution patterns and exposure concentrations of bioaerosols in industries suspected to have high levels of bioaerosol exposure. Methods: We selected 11 plants including 3 livestock feed plants (LF industry), 3 metal working fluids handling plants (MWFs industry), and 5 sawmills and measured total airborne bacteria, fungi, endotoxins, as well as dust. Airborne bacteria and fungi were measured with one stage impactor, six stage cascade impactor, and gelatin filters. Endotoxins were measured with polycarbonate filters. Results: The geometric means (GM) of the airborne concentrations of bacteria, fungi, and endotoxins were 1,864, $2,252\;CFU/m^3$, and $31.5\;EU/m^3$, respectively at the sawmills, followed by the LF industry (535, $585\;CFU/m^3$, and $22.0\;EU/m^3$) and MWFs industry (258, $331\;CFU/m^3$, and $8.7\;EU/m^3$). These concentrations by industry type were significantly statistically different (p < 0.01). The ratio of indoor to outdoor concentration was 6.2, 1.9, 3.2, and 3.2 for bacteria, fungi, endotoxins, and dust in the LF industry, 5.0, 0.9, 2.3, and 12.5 in the MWFs industry, and 3.7, 4.1, 3.3, and 9.7 in sawmills. The respiratory fractions of bioaerosols were differentiated by bioaerosol types and industry types: the respiratory fraction of bacteria in the LF industry, MWF industry, and sawmills was 59.4%, 72.0%, and 57.7%, respectively, and that of fungi was 77.3%, 89.5%, and 83.7% in the same order. Conclusion: We found that bioaerosol concentration was the highest in sawmills, followed by LF industry facilities and MWFs industry facilities. The indoor/outdoor ratio of microorganisms was larger than 1 and respiratory fraction of microorganisms was more than 50% of the total microorganism concentrations which might penetrate respiratory tract easily. All these findings suggest that bioaerosol in the surveyed industries should be controlled to prevent worker respiratory diseases.

Vitamin D maintains E-cadherin intercellular junctions by downregulating MMP-9 production in human gingival keratinocytes treated by TNF-α

  • Oh, Changseok;Kim, Hyun Jung;Kim, Hyun-Man
    • Journal of Periodontal and Implant Science
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    • 제49권5호
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    • pp.270-286
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    • 2019
  • Purpose: Despite the well-known anti-inflammatory effects of vitamin D in periodontal health, its mechanism has not been fully elucidated. In the present study, the effect of vitamin D on strengthening E-cadherin junctions (ECJs) was explored in human gingival keratinocytes (HGKs). ECJs are the major type of intercellular junction within the junctional epithelium, where loose intercellular junctions develop and microbial invasion primarily occurs. Methods: HOK-16B cells, an immortalized normal human gingival cell line, were used for the study. To mimic the inflammatory environment, cells were treated with tumor necrosis factor-alpha ($TNF-{\alpha}$). Matrix metalloproteinases (MMPs) in the culture medium were assessed by an MMP antibody microarray and gelatin zymography. The expression of various molecules was investigated using western blotting. The extent of ECJ development was evaluated by comparing the average relative extent of the ECJs around the periphery of each cell after immunocytochemical E-cadherin staining. Vitamin D receptor (VDR) expression was examined via immunohistochemical analysis. Results: $TNF-{\alpha}$ downregulated the development of the ECJs of the HGKs. Dissociation of the ECJs by $TNF-{\alpha}$ was accompanied by the upregulation of MMP-9 production and suppressed by a specific MMP-9 inhibitor, Bay 11-7082. Exogenous MMP-9 decreased the development of ECJs. Vitamin D reduced the production of MMP-9 and attenuated the breakdown of ECJs in the HGKs treated with $TNF-{\alpha}$. In addition, vitamin D downregulated $TNF-{\alpha}$-induced nuclear factor kappa B ($NF-{\kappa}B$) signaling in the HGKs. VDR was expressed in the gingival epithelium, including the junctional epithelium. Conclusions: These results suggest that vitamin D may avert $TNF-{\alpha}$-induced downregulation of the development of ECJs in HGKs by decreasing the production of MMP-9, which was upregulated by $TNF-{\alpha}$. Vitamin D may reinforce ECJs by downregulating $NF-{\kappa}B$ signaling, which is upregulated by $TNF-{\alpha}$. Strengthening the epithelial barrier may be a way for vitamin D to protect the periodontium from bacterial invasion.

제1기 비소세포폐암 환자의 수술적 절제 후 Matrix Metalloprotainase-2 활성도에 따른 재발 및 예후 (Activity of Matrix Metalloproteinase-2 and its Significance after Resection of Stage I Non-small Cell Lung Cancer)

  • 김상희;홍영숙;이진선;손대순;임유성;송인승;이혜숙;김도훈;김진국;최용수
    • Journal of Chest Surgery
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    • 제38권1호
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    • pp.38-43
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    • 2005
  • 기질금속단백분해효소-2 (MMP-2)는 기저막과 세포 외 기질의 분해에 작용하여 악성종양의 국소침윤 및 원격전이에 중요한 역할을 담당한다. 본 연구는 수술적인 절제를 시행 받은 제1기 비소세포폐암에서 MMP-2의 활성도를 측정하고 예후인자로서의 의미를 분석하고자 하였다. 대상 빛 방법: 수술 전 방사선 또는 항암제 요법을 시행하지 않은 병리학적 제1기 비소세포폐암 환자 34명을 대상으로 하였다. 폐엽 절제 조직에서 종양 조직과 비종양 조직의 단백질을 각각 추출하여 젤라틴-기질-단백분해효소 분석법을 시행하였다. 결과: MMP-2 활성도는 비종양 조직보다 종양 조직에서 높았다. 종양 조직, 비종양 조직 모두에서 비재발군에 비해 재발군의 MMP-2활성도가 높았으나 비종양 조직에서의 차이가 통계적 유의성이 있었다(p<0.01). 생존율 분석에서도 MMP-2 활성도가 높을 경우 불량한 생존을 보였으며 역시 비종양 조직에서 통계적인 의미가 있었다(p<0.01). 결론: 제1기 비소세포 페암 환자의 절제 폐엽 조직 중 비종양 조직에서의 MMP-2 활성도는 재발 및 생존과 연관성이 있으며 예후 인자로서의 가능성이 있다.