• BMB Reports
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• v.37 no.2
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• pp.199-205
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• 2004

The six lumbrokinase fractions (F1 to F6) with fibrinolytic activities were purified from earthworm Lumbricus rubellus lysates using the procedures of autolysis, ammonium sulfate fractionation, and column chromatography. The proteolytic activities on the casein substrate of the six iso-enzymes ranged from 11.3 to 167.5 unit/mg with the rank activity orders of F2 > F1 > F5 > F6 > F3 > F4. The fibrinolytic activities of the six fractions on the fibrin plates ranged from 20.8 to 207.2 unit/mg with rank orders of F6 > F2 > F5 > F3 > F1 > F4. The molecular weights of each iso-enzyme, as estimated by SDS-PAGE, were 24.6 (F1), 26.8 (F2), 28.2 (F3), 25.4 (F4), 33.1 (F5), and 33.0 kDa (F6), respectively. The plasminogen was activated into plasmin by the enzymes. The optimal temperature of the six iso-enzymes was $50^{\circ}C$, and the optimal pH ranged from pH 4-12. The four iso-enzymes (F1-F4) were completely inhibited by PMSF. The two enzymes (F5 and F6) were completely inhibited by aprotinin, TLCK, TPCK, SBTI, LBTI, and leupeptin. The N-terminal amino acid (aa) sequences of the first 20 to 22 residues of each fraction had high homology. All six isoenzymes had identical aa residues 2-3 and 13-15. The N-terminal 21-22 aa sequences of the F2, F3, and F4 isoenzymes were almost the same. The N-terminal aa sequences of F5 and F6 were identical.

• Biomedical Science Letters
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• v.15 no.1
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• pp.93-96
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• 2009

House dust mites (HDMs) play an important role in the occurrence of allergic diseases such as asthma and atopic dermatitis. Dermatophagoides pteronyssinus (Der p) is one of the most prevalent HDMs. It mediates the activation of T cells and monocytes, and induces the elevation of immunoglobulin E levels in allergic diseases. However, the effects of Der p on human monocytes have not been fully understood. In the present study, we investigated whether or not Der p has a great effect on the chemotactic activity of the human monocytic cell line, THP-1 cells, as induced by CC chemokines. We also show that the Der p extract (DpE) increased the chemotactic activity of THP-1 cells in response to MCP-1, RANTES, MIP-1${\alpha}$, and TARC, but had no effect on the expressions of CC chemokine receptors (CCRs) binding to CC chemokines in THP-1 cells. Protease inhibitors, such as aprotinin and E64, blocked the increased chemotaxis, while cytoplasmic $Ca^{2+}$ influx mediated by these chemokines was inhibited by DpE. These results indicate that DpE increases the chemotactic activity of THP-1 cells in response to CC chemokines by regulating the cells' protease-dependent mechanism. This finding may be useful in identifying the pathogenesis of allergic diseases induced by Der p.

• Archives of Pharmacal Research
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• v.28 no.7
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• pp.816-822
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• 2005

Catharsius protease-2 (CPM-2) was isolated from the body of dung beetles, Catharsius molossus, using a three step purification process (ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, and affinity chromatography on DEAE Affi-Gel blue). The purified CPM-2, having a molecular weight of 24 kDa, was assessed homogeneously by SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of CPM-2 was composed of X Val Gin Asp Phe Val Glu Glu lie Leu. CPM-2 was inactivated by $Cu^{2+}\;and\;Zn^{2+}$ and strongly inhibited by typical serine proteinase inhibitors such as TLCK, soybean trypsin inhibitor, aprotinin, benzamidine, and ${\alpha}_1$-antitrypsin. However, EDTA, EGTA, cysteine, $\beta$-mercaptoethanol, E64, and elastatinal had little effect on enzyme activity. In addition, antiplasmin and antithrombin III were not sensitive to CPM-2. Based on the results of a fibrinolytic activity test, CPM-2 readily cleaved $A{\alpha}-$ and $B{\beta}$-chains of fibrinogen and fibrin, and y-chain of fibrinogen more slowly. The nonspecific action of the enzyme resulted in extensive hydrolysis, releasing a variety of fibrinopeptides of fibrinogen and fibrin. Polyclonal antibodies of CPM-2 were reactive to the native form of antigen. The ELISA was applied to detect quantities, in nanograms, of the antigen in CPM-2 protein.

• The Journal of the Korean dental association
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• v.52 no.4
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• pp.218-233
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• 2014

Bispbosphonates are a class of pharmaceutic agents, which induce apoptosis of osteoclast as well as impair osteoclastic activity to suppress bone resorption. Thus, bisphophonates are effectively used to treat osteoporosis, multiple myeloma and to prevent bone metastases of malignant cancer. However, recently dental disease have been reported associated with Bisphosphonates. Thus, there are a number of discussions about proper prevention and treatment of bisphosphonate-related osteonecrosis of jaw(BRONJ). Marshall R. Urist in 1965 made the seminal discovery that a specific protein, BMP(bone morphogenetic protein), found in the extracellular matrix of demineralized bone could induce bone formation newly when implanted in extraosseous tissues in a host. BMPs are multi-functional growth factors which are members of the transforming growth factor-beta super family and their ability is that plays a pivotal roll in inducing bone. About 18 BMP family members have been identified and characterized. Among of them, BMP-2 and BMP-7 have significant importance in bone development. In this study, patients of BRONJ were recieved who visited Department of oral and maxillofacial surgery, school of dentistry, Wonkwang university for past 3 years from 2011 to 2013. We focused on the results of the surgical intervention. We suggest that new strategy of treatment used to rhBMP-2 and LFA(Lidocaine-Fibrinogen-Aprotinin)-collagen scaffold for patients of BRONJ. The purpose of this paper is to give a brief overview of BMPs and to critically review the clinical data currently available on rhBMP-2 and LFA collage scaffold.

• Journal of Chest Surgery
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• v.39 no.10 s.267
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• pp.770-774
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• 2006

Background: Continuous air leakage through chest tube after lung surgery may increase pt's hospital stay and lead to many complications including empyema etc. Chemical pleurodesis has frequently been used for prevention of air leakage. Therefore, we performed chemical pleurodesis using diluted fibrin glue in patients with continuous air leak-age and observed the effects and efficiency of treatment. Material and Method: From September, 2001 to August, 2005, 16 patients whose continuous air leakage lasted more than 7 days underwent chemical pleurodesis with diluted fibrin glue. The effects of treatment, complications and recurrences were reviewed. Dissolved fibrinogen 1.0 g and aprotinin 500,000 KIU were mixed in a 50 cc syringe (Mixed solution A). And dissolved thrombin 5,000 IU and Calcium chloride 600 mg were mixed in a 50 cc syringe (Mixed solution B). Cefazolin 1.0 g was mixed in a 50 cc syringe (Mixed solution C). Rubber tube was inserted between the chest tube and the collecting bottle. An inserted rubber tube was positioned 60cm above the patient and forming a loop appearance was done. Mixed solutions A, B and C were injected into the highest rubber tube. Results: Continuous air leakages disappeared in all f6 patients at next day. Chest tubes were removed after 3 days in all patients. Complications were chest pain in 12 patients (75%), leukocytosis in 14 patients (88%), fever and chill in 14 patients (88%). All complications were transient and disappeared without specific treatment. Conclusion: Our findings demonstrated that diluted fibrin glue chemical pleurodesis was effective in patients with continuous air leakage lasting more than 7 days. Diluted fibrin glue chemical pleurodesis had good results with acceptable complications. long term follow-up is necessary to evaluate the accurate effects of treatment and recurrence in a large number of patients.

• Korean Journal of Veterinary Research
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• v.43 no.3
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• pp.383-392
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• 2003

Insulin-like growth factors (IGFs) and their binding proteins (IGFBPs) are important regulators on the development of maternal tissues during pregnancy. This study was performed to examine the relationship between maternal IGFs/IGFBPs system (i.e: IGF-I, II, their receptors, and IGFBPs) in pre- and post-partum rats. The liver and kidney are important organs for the synthesis of IGFs and IGFBPs in adults. The levels of materanal IGFs and IGFBPs in serum, liver, and kidney were examined at 14 and 21 days of gestation and at 3, 7, 11, and 14 days after birth. The expression of IGFs and their receptors mRNA was also examined in fetal and maternal rat liver, kidney. IGF-I concentrations in maternal serum and liver were decreased during pregnancy. However, IGF-I concentration in maternal kidney was increased, having maximal effect at 14 days of gestation. IGF-I concentrations were decreased in serum, liver, and kidney of postpartum rat, compared to control (p < 0.05). On the other hand, IGF-II concentrations in serum, liver, and kidney were increased during pregnancy (p<0.05) and gradually decreased to control level in postpartum period. The levels of IGFBP-3 and IGFBP-2 are expressed in serum, liver, and kidney. However, IGFBP-3 is mainly expressed in serum and liver, and IGFBP-2 in kidney. The levels of IGFBP-3 and IGFBP-2 in maternal serum were markedly decreased during pregnancy and gradually recovered to control level during postpartum period by western ligand blotting. However, there was no change of IGFBP-3 and IGFBP-2 levels by western immunoblotting. The levels of IGFBP-3 and IGFBP-2 in maternal liver and kidney also showed the same pattern of serum, although the main IGFBP is different. In normal rat serum, IGF-I 150 kDa and 50 kDa carrier proteins were detected. The level of IGF-I 150 kDa carrier proteins in pregnant rat was decreased compared to normal rat, but that of 50 kDa carrier proteins was increased. IGFBP-3 protease activity was identified in pregnant rat serum and maternal placenta, and it was inhibited by EDTA ($Ca^{2+}$ chelating agent) and aprotinin (serine proteinase inhibitor). Taken together, these results suggest that the changes of IGFs and IGFBPs in maternal rats are regulated by liver and kidney IGFs and their receptors mRNA during the pregnancy.