• 한국식품영양과학회지
• /
• 제26권2호
• /
• pp.285-290
• /
• 1997

이상의 결과를 요약하연, 혈당값은 정상대조군 78{\pm}6.20mg/dl, 인슐린 비의존형 당뇨병 환자 $184{\pm}18.94mg/dl$로 인슐린 비의존형 당뇨병 환자의 혈당값이 유의적으로 높았다. IGF-I의 수준은 정상대조군 $9.32{\pm}1.48{\mu}g/ml$, 인슐린 비의존형 당뇨병 환자 $4.19{\pm}0.84{\mu}g/ml$로 인슐린 비의존형 당뇨병 환자의 IGF-I 수준이 유의적으로 낮았다. 인슐린 비의존형 당뇨병 환자의 혈청 중 IGFBP-1, -2는 증가한 반면, IGFBP-3, -4, -5는 감소하였다. IGFBP-3의 분해에 관여하는 효소는 PMSF, aprotinin, EDTA에 의해 저해되어 metal-dependent serine proteases임을 확인하였고, 효소의 크기가 대략 97,66kDa 이었으며 효소활성이 인슐린 비의존형 당뇨병 환자에게서 큼을 확인하였다.

• Maxillofacial Plastic and Reconstructive Surgery
• /
• 제13권2호
• /
• pp.129-136
• /
• 1991

Fibrin Sealants 는 과거 간, 비장, 신장등의 일반외과 수술에서 단순한 외과적 봉합으로는 해결하지 못하는 넓은 면적의 조직의 유착을 위하여 1940 년경 부터 개발되기 시작한 것으로 악안면 영역에서는 1970 년대 중반부터 미세신경접합술과 피부이식을 위하여 사용되기 시작한 후, 현재, 골이식후의 골세편의 고정, 혈관봉합술, 연조직에서 조직들의 유착과 지혈, 그리고 혈관종의 치유등을 목적으로도 광범위하게 연구되고 사용되고 있다. 이것은 인체에서 채취한 혈액응고인자 XIII 을 포함하는 Fibrinogen 성분과, 소에서 추출한 Thrombin 의 두가지 주요 성분으로 구성되며, Fibrinogen 용해제인 Aprotinin 액과 Thrombin 용해제인 염화칼슘액과 함께 네부분으로 구성된다. 각제품에 따라 그리고 사용된 농도에 따라 차이는 있으나, 대개 수분후에 조직이 응고되어 달라붙기 시작하고, 수시간후에 최대접착효과에 도달하며, 응고된 접착효과는 12 일에서 15 일간 유지되고 그후 정상적인 섬유소분해작용과 식세포활동에 의하여 분해된다. 저자는최근 6 개월간 서울대학교병원 구강악안면외과에서 28 명의 각종 질환 및 기형 환자에서 미세신경봉합술, 피부이식, 악교정성형술과 구개파열 또는 하악골 재건을 요하는 환자의 골이식후의 골세편의 고정, 경부곽청술이나 종양제거술후 각종 피부판 또는 근피판을 이용한 연조직의 적합, 혈관종의 처치, 후이개접근 법에 의한 악관절수술후의 외이도의 접합등 다양한 목적을 위하여 적용된 Fibrin Sealants 를 사용하여 양호한 결과를 얻었기에 보고하는 바이다.

• 한국토양동물학회지
• /
• 제4권2호
• /
• pp.101-106
• /
• 1999

지렁이 꼬리재생 초기에 발현되는 피브리노겐 분해효소의 활성은 세포외기질의 재구성에 연관되어 있을 것으로 생각된다. 본 연구에서는 지렁이의 꼬리 재생 중에 발현된 피브리노겐 분해효소의 활성과 특성을 밝히고자 하였다. 지렁이의 꼬리 재생 중에 발현되는 피브리노겐 분해효소는 최소 7개이며 그 분자량은 각각 58, 45, 32, 27, 23, 19 그리고 12kDa로 측정되었다. 피브리노겐 분해효소의 활성은 절단 후 1일부터 효소의 활성이 나타나서 7일까지 거의 유사한 정도의 활성이 유지되었으며, 7일 이후부터는 이 효소들의 활성이 급격히 감소하여 14일에는 그 활성이 대조군 수준으로 회복되었다. 지렁이 꼬리 재생 중 발현된 모든 피브리노겐 분해효소는 PMSF와 aprotinin를 처리하였을 때 효소들의 활성이 억제되었다. 또한 Ca$^{2+}$의 제거는 이효소의 활성에 영향을 미치지 않는 것으로 나타났다. 이러한 결과를 통하여 지렁이 꼬리재생시 발현되는 피브리노겐 분해효소는 serine계 단백질 분해효소임을 알 수 있었다.

• Archives of Pharmacal Research
• /
• 제20권1호
• /
• pp.34-38
• /
• 1997

Recombinant human epidermal growth factor (rhEGF), a polypeptide of 53 amino acid residues, is subject to degradation by numerous enzymes, especially proteases, when it is applied on the skin for the treatment of open wound. Amastatin, aprotinin, bestatin, EDTA, EGTA, gabexate, gentamicin, leupeptin, and TPCK were investigated for the possible protease inhibitors, which may use to protect rhEGF from degradation by the enzymes in the skin. Skin homogenates containing protease inhibitors and rhEGF were incubated at $37^{\circ}C$ for 30 minutes. After the reaction was stopped with trifluoroacetic acid, the amount of rhEGF remaining in the sample was determined with an HPLC method. The percentages of rhEGF degraded, at the skin/PBS ratio of 0.25, in the mouse, rat, and human skin homogenate were 85%, 70%, and 46%, respectively. The degree of degradation of rhEGF in the cytosolic fraction was higher than that in the membrane fraction and these enzyme reactions were completed in 30 minutes. Bestatin, EGTA, and TPCK showed significant inhibitory effects on the degradation of rhEGF in the two fractions (p<0.05), while the other protease inhibitors had no significant inhibitory effects or, even resulted in deleterious effects. Therefore, the formulation containing one or several inhibitors among these effective inhibitors would be a promising topical preparation of rhEGF for the treatment of open wound.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제8권3호
• /
• pp.187-191
• /
• 2003

Proteolytic enzymes existing in plant cell cultured media are the major reason for the loss of secreted human granulocyte-macrophage colony-stimulating factor (hGM-CSF). The addition of pepstatin, aprotinin and PMSF relatively decreased the proteolytic degradation of hGM-CSF in a conditioned medium, but sufficient prevention against the proteolytic activity could not be obtained with chemical protease inhibitors. Gelatin, as a competitive substrate for protease, showed a stabilizing effect in a conditioned medium. Compared to the initial hGM-CSF concentration in a conditioned medium. with 10 g/L of gelatin, 68% of the hGM-CSF remained after 5 days. In a cell culture experiment, 5 g/L of gelatin significantly stimulated the hGM-CSF production and accumulation in culture media, with no growth inhibition. compared to the controls (4.72 $\mu\textrm{g}$/L), the extracellular hGM-CSF level could be increased to 39.78 $\mu\textrm{g}$/L with the addition of 5 g/L of gelatin.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• 제49권4호
• /
• pp.208-213
• /
• 2023

Objectives: Orthognathic surgery is a corrective intervention for maxillofacial deformities. Bleeding is a major concern for oral and maxillofacial surgeons. Various agents, such as hemocoagulase, tranexamic acid, and aprotinin have been developed to reduce intraoperative bleeding and transfusion requirements. Therefore, in this study we aimed to investigate the effects of hemocoagulase and tranexamic acid, as well as their simultaneous use, to reduce bleeding during orthognathic surgery. Patients and Methods: This retrospective study included patients who had undergone simultaneous orthognathic surgery of the maxilla and mandible between January 2013 and September 2022 and were classified into three groups based on drugs administered: hemocoagulase (Botropase), tranexamic acid, and a combination of both drugs. We recorded patient age, sex, weight, blood loss, and duration of surgery. Red blood cell (RBC), hemoglobin, hematocrit, and platelet levels were measured before, immediately after, and one day after surgery. Results: No statistically significant differences were found in blood loss, RBC, hemoglobin, hematocrit, or platelet levels between any of the groups. There were no differences in the drug effects between Le Fort I and bilateral mandibular sagittal split osteotomies, with or without double genioplasty. However, there were significant reductions in RBC, hemoglobin, hematocrit, and platelet levels during genioplasty. Conclusion: Tranexamic acid, hemocoagulase, and their combination had similar efficacy in patients who underwent Le Fort I and bilateral mandibular sagittal split osteotomies with and without genioplasty.

• Journal of Microbiology and Biotechnology
• /
• 제24권2호
• /
• pp.245-253
• /
• 2014

A fibrinolytic enzyme was produced by an edible mushroom of Pleurotus ostreatus using submerged culture fermentation. The enzyme was purified from the culture supernatant by applying a combination of freeze-thaw treatment, ammonium sulfate precipitation, hydrophobic interaction, and gel filtration chromatographies. The enzyme was purified by a 147-fold, with a yield of 7.54%. The molecular masses of the enzyme an determined by gel filtration and SDS-PAGE were 13.6 and 18.2 kDa, respectively. The isoelectric point of the enzyme was 8.52. It hydrolyzed fibrinogen by cleaving the ${\alpha}$ and ${\beta}$ chains of fibrinogen followed by the ${\gamma}$ chains, and also activated plasminogen into plasmin. The enzyme was optimally active at $45^{\circ}C$ and pH 7.4. The enzyme activity was completely inhibited by EDTA, whereas protease inhibitors of TPCK, SBTI, PMSF, aprotinin and pepstatin showed no inhibition on its activity. The partial amino acid sequences of the enzyme as determined by Q-TOF2 were ATFVGCSATR, GGTLIHESSHFTR, and YTTWFGTFVTSR. These sequences showed a high degree of homology with those of metallo-endopeptidases from P. ostreatus and Armillaria mellea. The purified enzyme can also be applied as a natural agent for oral fibrinolytic therapy or prevention of thrombosis.

• Journal of Microbiology and Biotechnology
• /
• 제16권2호
• /
• pp.264-271
• /
• 2006

Two fibrinolytic enzymes were purified from the culture supernatant of Fomitella fraxinea mycelia by ion-exchange and gel filtration chromatographies, and were designated as F. fraxenia proteases 1 and 2 (FFP1 and FFP2). The apparent molecular masses of the enzymes were estimated to be 32 kDa and 42 kDa, respectively, by SDS-PAGE and gel filtration chromatography. Both enzymes had the same optimal temperature ($40^{\circ}C$), but different pH optima (10.0 and 5.0 for FFP1 and FFP2, respectively). FFP1 was relatively stable at pH 7.0-9.0 and temperature below $30^{\circ}C$, whereas FFP2 was very stable in the pH range of 4-11 and temperature below $40^{\circ}C$. FFPI activity was completely inhibited by phenylmethylsulfonyl fluoride (PMSF) and aprotinin, indicating that this enzyme is a serine protease. The activity of FFP2 was enhanced by the addition of $CO^{2+}$ and $Zn^{2+}$ and inhibited by $Cu^{2+},\;Ni^{2+}$, and $Hg^{2+}$. Furthermore, FFP2 activity was strongly inhibited by EDTA and 1,10-phenanthroline, implying that the enzyme is a metalloprotease. Both enzymes readily hydrolyzed fibrinogen, preferentially digesting the $A{\alpha}$- and $B{\beta}$-chains of fibrinogen over ${\gamma}$-chain. FFP1 showed broad substrate specificity for synthetic substrates, but FFP2 did not. $K_{m}$ and $V_{max}$ values of FFP1 for a synthetic substrate, N-succinyl-Ala-Ala-Pro-Phe-pNA, were 0.213 mM and 39.68 units/ml, respectively. The first 15 amino acids of the N-terminal sequences of both enzymes were APXXPXGPWGPQRIS and ARPP(G)VDGQ(R,I)SK(L)ETLPE, respectively.

• 대한한방내과학회지
• /
• 제20권1호
• /
• pp.198-209
• /
• 1999

A thrombus is a mass formed from the constituents of the blood within the vessels or the heart during life. The process of its formation is known as thrombosis, It has been generally accepted that Holotrichia is an useful medicine for thrombosis. The rate of fibrinolysis of Holotrichia extracts increase as incubation times. Especially 52 days, the effect on the extracts has an maximum increased fibrinolytic activity, Heat-and-pH-stability of the extract on fibrinolysis is relative to temperature, At $37^{\circ}C$, it has activating effect between pH 4 and pH 12. At higher temperature, especially $80^{\circ}C$, an excessive increase in temperature has a deactivating effect on the extracts. Optimal pH of the extract on fibrinolysis is between pH 7.0 and pH 8.5, it is effective within a relatively broad pH range. In experiments of various inhibitors of Holotrichia extracts fibrinolytic activity, there are strong inhibitive effect on SBTI and Aprotinin, and a few inhibitive effect on DFP and t-AMCHA, no effect on PSMB and TLCK. Holotrichia extracts mixing with fibrinogen are observed Electron microscopy. it shows partially erosive-shaped fibrinolytic activity. In a SDS-PAGE of the extract having the fibrinolytic activity, three bands are found, protein 1, 2 and 3 having a molecular weight of 30,000, 45,000 and 60,000 Dalton.

• Journal of Microbiology and Biotechnology
• /
• 제21권3호
• /
• pp.256-262
• /
• 2011

A 20.9-kDa metalloprotease was isolated from dried fruiting bodies of the wild basidiomycete mushroom Lepista nuda. The N-terminal amino acid sequence of the protease was seen to be ATFVLTAATNTLFTA, thus displaying no similarity with the sequences of previously reported metalloproteases. The protease was purified using a procedure that entailed ion-exchange chromatography on CM-Cellulose, Q-Sepharose, and Mono S, and FPLC-gel filtration on Superdex 75. The protease functioned at an optimum pH of 7.0 and an optimum temperature of $50^{\circ}C$. It was also noted that the protease demonstrated a proteolytic activity of 1,756 U/mg toward casein. The $K_m$ of the purified protease toward casein was 6.36 mg/ml at a pH of 7.0 and with a temperature of $37^{\circ}C$, whereas the $V_{max}$ was 9.11 ${\mu}g\;ml^{-1}\;min^{-1}$. The activity of the protease was adversely affected by EDTA-2Na, suggesting that it is a metalloprotease. PMSF, EGTA, aprotinin, and leupeptin exerted no striking inhibitory effect. The activity of the protease was enhanced by $Fe^{2+}$, but was curtailed by $Cd^{2+}$, $Cu^{2+}$, $Hg^{2+}$, $Pb^{2+}$, $Zn^{2+}$, and $Fe^{2+}$ ions. The protease also exhibited inhibitory activity against HIV-1 reverse transcriptase with an $IC_{50}$ value of 4.00 ${\mu}M$. The $IC_{50}$ values toward hepatoma Hep G2 and leukemia L1210 cells in vitro were 4.99 ${\mu}M$ and 3.67 ${\mu}M$, respectively.