No abstract available.
Animals ; Female ; Ovary* ; Rats* ; Serine Proteases* ; Serine*

Kunitz-type serine protease inhibitors are a class of ubiquitous protease inhibitors, which play important roles in various life activities. The structures of such inhibitors are generally stable, and are usually characterized by the presence of one or several Kunitz domains in tandem, which are able to bind to serine proteases in a manner similar to substrate binding, thereby inhibiting enzyme activity. In terms of function, Kunitz-type serine protease inhibitors are involved in processes such as blood coagulation and fibrinolysis, tumor immunity, inflammation regulation, and resistance to bacterial and fungal infections. This article summarizes the advances of Kunitz-type serine protease inhibitors and provides new ideas for the development of novel Kunitz-type serine protease inhibitors.
Protease Inhibitors ; Serine Proteases ; Serine Proteinase Inhibitors

BACKGROUND: Mucus hypersecretion in the patients with airway diseases represents poor prognosis as well as discomfort. However, there is no known therapy for its effective control. One important component of mucus is mucin, a glycosylated protein, which endows mucus with viscosity. We studied whether a proteinase has a role in mucin secretion and if so, which. METHODS: (1) Inhibition of mucin secretion Group-specific proteinase inhibitors were tested to evaluate whether a proteinase belonging to a group of proteinases plays a role in mucin secretion. Phenylmethylsulfonyl fluoride(PMSF, a serine proteinase inhibitor), E-64(a cysteine proteinase inhibitor), Pepstatin(an aspartic proteinase inhibitor) and 1, 10-Phenanthroline(a metalloproteinase inhibitor) were treated into the Calu-3 cell line for 24 hours. The enzyme linked immunoabsorbant assay(ELISA) for MUC5AC was performed to evaluate the amount of mucin secretion and to compare with a control. (2) Stimulation of mucin secretion Matrix metalloproteinase-9(MMP-9), MMP-12 and TACE(TNF-alpha converting enzyme), which are known to be related with airway diseases, were used to be treated into Calu-3 for 24 hours. ELISA for MUC5AC was performed to evaluate the amount of mucin secretion and to compare with the controls. RESULTS: (1) PMSF(10(-4)M), E-64(10(-4)M), Pepstatin(10(-6)M) and 1, 10-Phenanthroline(10(-4)M) reduced the MUC5AC secretion by 1 +/- 4.9%(mean +/- standard deviation; P=1.0 compared with the control), -6 +/- 3.9%(P=0.34), -13 +/- 9.7%(P=0.34) and 41 +/- 8.2%(P=0.03), respectively. (2) The amounts of MUC5AC secretion stimulated by MMP-9(250ng/ml), MMP-12(100ng/ml) and TACE(200ng/ml) were 103 +/- 6%(P=0.39), 102 +/- 8%(P=1.0) and 107 +/- 13%(P=0.39), respectively, compared with the controls. CONCLUSION: Metalloproteinase(s) is (are) suggested to play a role in mucin secretion. It appears that metalloproteinases, other than MMP-9, MMP-12 or TACE, affect the mucin secretion in this in vitro model.
Cell Line ; Cysteine Proteases ; Enzyme-Linked Immunosorbent Assay ; Humans ; Metalloproteases ; Mucins* ; Mucus ; Peptide Hydrolases ; Prognosis ; Serine Proteases ; Viscosity

PURPOSE: Fusarium species are among prevalent airborne fungi and causative agents of human respiratory atopic disorders. We previously identified a 36.5-kDa F. proliferatum component recognized by IgE antibodies in 9 (53%) of the 17 F. proliferatum-sensitized atopic serum samples. The purpose of this study is to characterize the 36.5-kDa allergen of F. proliferatum. METHODS: Characterization of allergens and determination of IgE cross-reactivity were performed by cDNA cloning/expression and immunoblot inhibition studies. RESULTS: Based on the finding that the 36.5-kDa IgE-binding component reacted with the mouse monoclonal antibody FUM20 against fungal vacuolar serine protease allergens, the cDNA of F. proliferatum vacuolar serine protease (Fus p 9.0101) was subsequently cloned. Nine serum samples from respiratory atopic patients with IgE binding to the vacuolar serine protease allergen of Penicillium chrysogenum (Pen ch 18) also showed IgE-immunoblot reactivity to rFus p 9.0101. The purified rFus p 9.0101 can inhibit IgE and FUM20 binding to the 36.5-kDa component of F. proliferatum. Thus, a novel and important Fus p 9.0101 was identified. The rPen ch 18 can inhibit IgE binding to Fus p 9.0101. It indicates that IgE cross-reactivity between Fus p 9.0101 and Pen ch 18 also exists. Furthermore, neither rFus p 9.0101 K88A nor rPen ch 18 K89A mutants inhibited IgE binding to rFus p 9.0101. Lys88 was considered a critical core amino acid in IgE binding to r Fus p 9.0101 and a residue responsible for IgE cross-reactivity between Fus p 9.0101 and Pen ch 18 allergens. CONCLUSIONS: Results obtained from this study indicate that vacuolar serine protease may be a major allergen of F. proliferatum and an important IgE cross-reactive pan-fungal allergen, and provide important bases for clinical diagnosis of fungal allergy.
Allergens ; Animals ; Antibodies ; Clone Cells ; Diagnosis ; DNA, Complementary ; Fungi ; Fusarium* ; Humans ; Hypersensitivity ; Immunoglobulin E ; Mice ; Penicillium chrysogenum ; Serine Proteases* ; Serine*

OBJECTIVE: The purposes of this study were to examine the expression of matriptase, and its inhibitor, HAI-1, in epithelial ovarian cancer and to assign clinicopathological correlations and to discuss the matriptase/inhibitor (HAI-1) system in the context of ovarian cancer and to examine the possibility that this system might be a useful therapeutic target in this disease. METHODS: A retrospective study was performed in 51 patients with primary epithelial ovarian cancer staged over Ic who have been diagnosed and treated at Kyung Hee university medical center from Jan. 1991 to Mar. 2003. They were managed with cytoreductive surgery and chemotherapy. This study was performed in paraffin embedded blocks of primary epithelial ovarian cancer of 51 patients by means of immunohistochemistry. In addition, to validate protein expression data at the gene level, matriptase/HAI-1 mRNA expression was evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) on frozen specimens from 10 ovarian cancers. Statistical analyses of immunohistochemistry (IHC) expression data with clinicopathological parameters and survival were then performed. RESULTS: Of 51 ovarian tumors tested, 25 (49%) and 37 (72.5%) were positive staining for matriptase and HAI-1 by IHC, respectively. Of 18 stage I/II tumors, 11 (61.1%) stained positive for matriptase, and 15 (83.3%) stained positive for HAI-1; Of 18 stage I/II tumors, 10 (55.6%) tumors showed coexpression. Of 33 stage III/IV tumors, 14 (42.4%) stained positive for matriptase and 22 (66.7%) stained positive for HAI-1; Of 33 stage III/IV tumors, 11 (33.3%) tumors showed coexpression. CONCLUSION: No relationship was found between the expression of either matriptase or HAI-1 with clinicopathological parameters and survival. However, stage I/II ovarian tumors are more likely to express matriptase and HAI-1 than are the more advanced disease stage III/IV tumors. Correspondingly, the low frequency of matriptase and HAI-1 coexpression is more likely to be associated with stage III/IV tumors than stage I/II tumors. Such an imbalance in the matriptase: HAI-1 ratio could promote the proteolytic activity of matriptase and, consequently, a more invasive phenotype in the advanced tumors.
Academic Medical Centers ; Drug Therapy ; Humans ; Immunohistochemistry ; Ovarian Neoplasms* ; Paraffin ; Phenotype ; Retrospective Studies ; RNA, Messenger ; Serine Proteases* ; Serine*

10–30% of patients with pancreatitis can be categorized as idiopathic pancreatitis, and some of them may be due to genetic alterations. Since hereditary pancreatitis develops from pediatric patients with symptoms related to pancreatitis, which usually progresses to chronic pancreatitis around 30 years of age, special attention should be paid to the development of pancreatic cancer in such patients. Up to now, there have been more than 30 genetic alterations associated with pancreatitis. Alterations in protease serine 1 (PRSS1), serine protease inhibitor Kazal type 1 (SPINK1), cystic fibrosis transmembrane conductance regulator (CFTR) and chymotrypsin C (CTRC) are common, which show diversity according to race and region. It is important to understand the characteristics of Korean patients with idiopathic pancreatitis through genetic studies. The purpose of this article is to review the role of genetic variations in the pathophysiology of idiopathic pancreatitis and to survey the results of Korean studies of idiopathic pancreatitis.
Chymotrypsin ; Continental Population Groups ; Cystic Fibrosis Transmembrane Conductance Regulator ; Genetic Variation ; Humans ; Pancreatic Neoplasms ; Pancreatitis* ; Pancreatitis, Chronic ; Serine ; Serine Proteases

Recently, many works to treat chronic corneal ulcer have been progressed. It has been reported that the Aprotinin, one of them, is serine protease inhibitor and is useful to treat therapy-resistant chronic corneal ulcer because it decreases the plasmin level in tear fluid that was increased in corneal ulcer. We performed this study to evaluate the effect of Aprotinin to the reepithelization of cornea according to its concentration. We made corneal burn in rabbits and instilled topical antibiotics and Aprotinin 500u/ml and 200u/ml, four times a day. After instillation, we compared the process of corneal epithelial wound healing, according to the time interval, clinically and histopathologically in each group. As a result, wound healing of cornea treated with combination of antibiotics and Aprotinin was delayed rather than that treated with antibiotics only. And combination therapy with Aprotinin 500n/ml is more effective than with Aprotinin 2000n/ml. This data suggests that high concentration of Aprotinin alone is not helpful to tresat the therapyresistant chronic corneal ulcer.
Anti-Bacterial Agents ; Aprotinin* ; Burns* ; Cornea ; Corneal Ulcer ; Fibrinolysin ; Rabbits* ; Serine Proteases ; Wound Healing

Uremic pruritus is a common problem in patients with end-stage renal disease (ESRD), but the underlying mechanisms are not yet fully understood. We aimed to investigate the association between severity of uremic pruritus and cutaneous serine protease activity, as well as proteinase-activated receptor-2 (PAR-2) expression. Twelve ESRD patients with pruritus, 4 ESRD patients without pruritus, and 6 healthy controls were enrolled. Skin biopsies were obtained from the abdomen. Protease activity and PAR-2 expression in the epidermis were examined by in situ zymography and confocal laser microscopy, respectively. All ESRD patients presented more pronounced cutaneous protease activity compared with that in healthy controls. The skin samples from the patients with pruritus showed higher protease activity than either nonpruritic ESRD patients or healthy controls. The epidermis in all samples of ESRD patients presented higher immunoreactivity against PAR-2 versus those of healthy controls. In addition, correlation analysis between PAR-2 expression and VAS pruritus scores showed a significant positive correlation. Our data suggests that levels of serine protease and PAR-2 expression could play important roles in the pathogenesis of uremic pruritus.
Abdomen ; Biopsy ; Epidermis ; Humans ; Kidney Failure, Chronic* ; Microscopy, Confocal ; Pilot Projects* ; Pruritus* ; Serine Proteases ; Skin ; Uremia

PURPOSE:In patients with a higherrisk of bleeding, performing CVVH with heparin or saline anticoagulation is associated with increased bleeding or thrombotic risk. Nafamostat mesilate (NM), a serine proteinase inhibitor, while inhibiting various clotting factors in filter circuit, is characterized by short half life resulting in little systemic anticoagulation effect. Accordingly, we prospectively evaluated the anticoagulant effect and safety of NM in patients with a higher risk of bleeding who underwent CVVH. METHODS:Among 43 patients with high risk of bleeding [defined by (1) INR>2, aPTT>20 sec, platelet< 50,000/mm3 or (2) ongoing bleeding, major hemorrhage/surgery in the last 48 hrs], 20 patients were treated with continuous nafamostat mesilate infusion (10-20 mg/hr) and remaining 23 patients were treated with saline bolus infusion (100 mLq 1 hr) for CVVH anticoagulation. RESULTS:As compared with saline bolus group, mean circuit life was significantly longer in NM infusion group (28.73+/-12.67 versus 16.34+/-7.86, p=0.001). There was no significant bleeding complication in either saline bolus or NMinfusion group. In subgroup analysis according to the presence of abnormal coagulation status (defined by INR>2, aPTT>20 sec, platelet<50,000/mm3), the positive effect of NM on circuit lifespan persisted irrespective of the coagulation status. CONCLUSION:As compared with saline bolus, nafamostat mesilate infusion was associated with higher CVVH filter life. In patients with high risk of bleeding, nafamostat mesilate can be used as a safe and effective anticoagulant for CVVH with acceptable filter life
Guanidines ; Half-Life ; Hemofiltration ; Hemorrhage ; Heparin ; Humans ; Mesylates ; Prospective Studies ; Serine Proteases

BACKGROUND: Aprotinin is a potent, nonspecific broad serine protease inhibitor. It's inhibitory effects on intrinsic pathway of coagulation cascade can augment anticoagulation by heparin. This study designed to demonstrate augmented anticoagulation of aprotinin to heparin contaminated blood on thromboelastography(TEG). METHODS: This study designed into two phases for 21 healthy volunteers undergoing elective opeation. The first phase study, it was for looking at TEG differences between blood treated with aprotinin 200 KIU and blood treated with heparin 0.05 unit and 0.1 unit per blood 1 ml. The second phase study was for looking at anticoagulation of aprotinin added by heparin 0.05 unit and 0.1 unit per blood 1 ml and their reversal added by optimal dose of protamine sulfate. RESULTS: The aprotinin treated blood showed only a prolonged reaction time. Blood treated with incremental dose of heparin showed longer reaction time and smaller alpha angle than TEGs of native blood. Aprotinin added to the heparin contaminated blood showed much longer reaction time and much less alpha angle when compared with TEGs of aprotinin or heparin treated blood. Depressed TEG pattern by the heparin and aprotinin mixture reversed back to the TEGs of blood treated with aprotinin when optimal dose of protamine added. CONCLUSIONS: Those results suggest that aprotinin administered in open cardiac surgery can augment the remained anticoagulation effect due to heparin even after first dose fo protamine after weaning of cardiopulmonary bypass. This is of clinically improtance to distinguish heparin related coagulopathy from heparin non related coagulopathy by thromboelastography.
Aprotinin* ; Cardiopulmonary Bypass ; Healthy Volunteers ; Heparin* ; Protamines ; Reaction Time ; Serine Proteases ; Thoracic Surgery ; Thrombelastography* ; Weaning