• Title/Summary/Keyword: Proteolytic Enzymes

Search Result 245, Processing Time 0.025 seconds

Analysis of Evaluation Methods for the Efficacy of Protein Removal Agents for Soft Contact Lens (소프트콘택트렌즈 단백질제거제의 효능 평가법 분석)

  • Byuna, Hyun Young;Sung, Hyung Gyeong;Won, Hye Lim;Shim, Ji In;Park, Mijung;Kim, So Ra
    • Journal of Korean Ophthalmic Optics Society
    • /
    • v.19 no.1
    • /
    • pp.51-57
    • /
    • 2014
  • Purpose: The present study was conducted to establish the experimental condition for the proper evaluation of protein removal efficacy when developing protein removal agents. Its protein removal efficacy was further analyzed and compared with the result from protein removal efficacy against protein deposition on contact lens to suggest the evaluation method for efficacy of protein removal agents. Methods: Protein digestibility assay presented in the Korean pharmacopoeia was selected to establish the evaluation method for efficacy of papain, pancreatin, subtilisin A and protease itself as a ingredient and protein removal tablets or solution containing those enzymes and find a suitable test conditions. Furthermore, the cleaning efficacy of commercially available protein removal tablets and solution on balafilcon A lens deposited with protein artificially was measured and the correlation between two evaluation methods was further analyzed. Results: When pancreatin itself and the product containing pancreatin was evaluated by protein digestibility assay, both reached 28 IU/mg, the standard value of protein digestibility suggested by the Korean pharmacopoeia. In case of protease and subtilisin A tested with trichloroacetic acid B solution, both of them met the enzyme activity level proposed by the manufacturers when they were evaluated by protein digestibility assay however, papain and subtilisin A tested with trichloroacetic acid A solution were not reached the enzyme activity level. Among protein removal agents, three products except a product containing pancreatin did not meet the enzyme activity value specified by the manufacturer when they were evaluated by protein digestibility assay. However, actual protein removal efficacy of three products except a papain-containing product on the lens was greater than 90% protein removal. In the case of papain-containing protein removal product, its effect was not measured by protein digestibility assay however, its actual protein removal efficacy on the lens reached 73.72%. Conclusions: From the results, it was confirmed that the efficacy of protein removal agents for contact lens should be evaluated by different method according to the type of proteolytic enzyme contained. That is, the protein removal agents containing pancreatin, protease and subtilisin A can be evaluated by protein digestibility assay and protein removal efficiency evaluation and the products containing papain can be effectively evaluated by only the evaluation method for protein removal efficiency employing the lens.

PROCESSING OF DRILL SOLUBLE AND ITS AMINO ACID COMPOSITION (Krill solube의 가공 및 아미노산 조성)

  • LEE Eung-Ho;KIM Se-Kwon;CHO Duck-Jae;HAN Bong-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.12 no.4
    • /
    • pp.235-240
    • /
    • 1979
  • A study on the amino acid composition of raw frozen krill, and krill solubles manufactured in forms of paste and powder has been carried out. The raw frozen krill was thawed, chopped, mixed and homogenized with same amount of water. The mixture was autolyzed or hydrolyzed by tile addition of $0.2\%$ pronase-p, a commercial proteolytic enzyme, to the weight of the raw frozen krill at $45^{\circ}C$ for 4 hours. After a thermal inactivation of enzymes at $95^{\circ}C$ for 15 minutes, the autolysate and the hydrolysate were centrifuged and filtered through gauzes, respectively, and then tile lipid layer in the supernatant was removed, The autolysate and the hydrolysate were finally concentrated under reduced atmospheric pressure in a rotary vacuum evaporator at $45^{\circ}C$ for 1 hour to produce the krill solubles in form of paste. The powdered krill solubles were prepared by the addition of $5\%$ starch to the autolysate and hydrolysate and by means of concentration in the rotary vacuum evaporator at $45^{\circ}C$ for 30 minutes and a forced air drying at $58^{\circ}C$ for 3 hours with a air velocity of 3m/sec. Among the amino acids in raw frozen krill, glutamic acid, lysine, and aspartic acid showed high values in quantity and then followed leucine, alanine, arginine, glycine and proline. The qnantity of histidine was very small and that of cystine was only in trace. The krill solubles in forms of paste and powder prepared by autolysis and hydrolysis with pronase-p revealed almost the same patterns in amino acid composition as in raw frozen krill. In case of free amino acids, a large quantity of it in raw frozen krill consisted of lysine, arginine, proline, alanine and leucine. The quantities of cystine, histidine and glutamic acid were, in contrast, very small. In the soluble krill paste prepared by autolysis, lysine, leucine, threonine and alanine existed in large quantities among the free amino acids and cystine, aspartic acid and histidine existed in small quantities. The contents of almost all of the free amino acids ill soluble krill paste perpared by hydrolysis with pronase-p were increased slightly as compared with those in soluble krill paste prepared by autolysis. In this product, the contents of cystine, histidine and serine were very low and lysine, leucine, arginine and proline were the dominant group in quantities among the free amino acids. The krill solubles in forms of paste and powder were not inferior to whole egg in the view point of its essential amino acid composition.

  • PDF

Effect of the Addition of Tenderizers on Quality Characteristics of Intermediate Moisture-apple snail(Cipangopaludin chinensis malleasta) Products (연화제 첨가가 우렁이 중간수분식품의 품질 특성에 미치는 영향)

  • Yang, Han-Sul;Chun, Ji-Yeon;Kim, Byung-Chul;Kang, Sung-Won;Jeong, Chang-Ho;Heo, Ho-Jin;Cho, Sung-Hwan;Choi, Sung-Gil
    • Food Science and Preservation
    • /
    • v.14 no.3
    • /
    • pp.288-293
    • /
    • 2007
  • Intermediate moisture-apple snail products were prepared by adding glycerol, sorbitol, kiwi, or pineapple(2% or 5% w/w), as tenderizers, and by drying at $4^{\circ}C$ for 24 hr. The effects of the tenderizers on textural and sensory properties of the apple snail products at intermediate moisture levels were investigated. Moisture content and water activity of the products were ranged from 26.25 to 34.48% and from 0.83 to 0.87, respectively. The addition of glycerol significantly lowered water activity of apple snail samples compared to control prepared without tenderizers. On the other hand, significant increases in moisture content and water activity were observed in apple snail samples treated with kiwi or pineapple(p<0.05). All apple snail samples treated with tenderizers showed a lower shear force than did the control. Apple snail samples treated with 5%(v/v) glycerol showed a higher equilibrium moisture content than did the other samples. SDS-PAGE indicated that proteolytic enzymes in kiwi and pineapple clearly changed the structure of the myosin heavy chain and actin filaments of myofibrillar protein in apple snail samples. Intermediate moisture apple snail samples treated with tenderizers showed significantly improved overall sensory characteristics. The highest overall acceptability was obtained from apple snail samples treated with 5% pineapple, while the lowest overall acceptability was noted in the control sample. This study demonstrates that an acceptable apple snail, with intermediate moisture content, may be produced by using tenderizers at appropriate concentrations.

The Significance of Plasma Urokinase-type Plasminogen Activator and Type 1 Plasminogen Activator Inhibitor in Lung Cancer (폐암에서 혈장 Urokinase-Type Plasminogen Activator 및 Type 1 Plasminogen Activator Inhibitor의 의의)

  • Park, Kwang-Joo;Kim, Hyung-Jung;Ahn, Chul-Min;Lee, Doo-Yun;Chang, Joon;Kim, Sung-Kyu;Lee, Won-Young
    • Tuberculosis and Respiratory Diseases
    • /
    • v.44 no.3
    • /
    • pp.516-524
    • /
    • 1997
  • Background : Cancer invasion and metastasis require the dissolution of the extracellular matrix in which several proteolytic enzymes are involved. One of these enzymes is the urokinase-type plasminogen activator(u-PA), and plasminogen activator inhibitors(PAI-1, PAI-2) also have a possible role in cancer invasion and metastasis by protection of cancer itself from proteolysis by u-PA. It has been reported that the levels of u-PA and plasminogen activator inhibitors in various cancer tissues are significantly higher than those in normal tissues and have significant correlations with tumor size and lymph node involvement. Here, we measured the concentration of plasma u-PA and PAI-1 antigens in the patients with lung cancer and compared the concentration of them with histologic types and staging parameters. Methods : We measured the concentration of plasma u-PA and PAI-1 antigens using commercial ELISA kit in 37 lung cancer patients, 21 benign lung disease patients and 24 age-matched healthy controls, and we compared the concentration of them with histologic types and staging parameters in lung cancer patients. Results : The concentration of u-PA was $1.0{\pm}0.3ng/mL$ in controls, $1.0{\pm}0.3ng/mL$ in benign lung disease patients and $0.9{\pm}0.3ng/mL$ in lung cancer patients. The concentration of PAI-1 was $14.2{\pm}6.7ng/mL$ in controls, $14.9{\pm}6.3ng/mL$ in benign lung disease patients, and $22.1{\pm}9.8ng/mL$ in lung cancer patients. The concentration of PAI-1 in lung cancer patients was higher than those of benign lung disease patients and controls. The concentration of u-PA was $0.7{\pm}0.4ng/mL$ in squamous cell carcinoma, $0.8{\pm}0.3ng/mL$ in adenocarcinoma, 0.9ng/mL in large cell carcinoma, and $1.1{\pm}0.7ng/mL$ in small cell carcinoma. The concentration of PAI-1 was $22.3{\pm}7.2ng/mL$ in squamous cell carcinoma, $22.6{\pm}9.9ng/mL$ in adenocarcinoma, 42 ng/mL in large cell carcinoma, and $16.0{\pm}14.2ng/mL$ in small cell carcinoma. The concentration of u-PA was 0.74ng/mL in stage I, $1.2{\pm}0.6ng/mL$ in stage II, $0.7{\pm}0.4ng/mL$ in stage IIIA, $0.7{\pm}0.4ng/mL$ in stage IIIB, and $0.7{\pm}0.3ng/mL$ in stage IV. The concentration of PAI-1 was 21.8ng/mL in stage I, $22.7{\pm}8.7ng/mL$ in stage II, $18.4{\pm}4.9ng/mL$ in stage IIIA, $25.3{\pm}9.0ng/mL$ in stage IIIB, and $21.5{\pm}10.8ng/mL$ in stage IV. When we divided T stage into T1-3 and T4, the concentration of u-PA was $0.8{\pm}0.4ng/mL$ in T1-3 and $0.7{\pm}0.4ng/mL$ in T4, and the concentration of PAI-1 was $17.9{\pm}5.6ng/mL$ in T1-3 and $26.1{\pm}9.1ng/mL$ in T4. The concentration of PAI-1 in T4 was significantly higher than that in T1-3. The concentration of u-PA was $0.8{\pm}0.4ng/mL$ in M0 and $0.7{\pm}0.3ng/mL$ in M1, and the concentration of PAI-1 was $23.6{\pm}8.3ng/mL$ in M0 and $21.5{\pm}10.8ng/mL$ in M1. Conclusions : The plasma levels of PAI-1 in lung cancer were higher than benign lung disease and controls, and the plasma levels of PAI-1 in T4 were significantly higher than T1-3. These findings suggest involvement of PAI-1 with local invasion of lung cancer, but it should be confirmed by the data on comparison with pathological staging and tissue level in lung cancer.

  • PDF

The Evaluation of IL-8 in the Serum of Pneumoconiotic patients (진폐증 환자에서의 혈청내 IL-8 농도)

  • Ahn, Hyeong Sook;Kim, Ji Hong;Chang, Hwang Sin;Kim, Kyung Ah;Lim, Young
    • Tuberculosis and Respiratory Diseases
    • /
    • v.43 no.6
    • /
    • pp.945-953
    • /
    • 1996
  • Background : Many acute and chronic lung diseases including pneumoconiosis are characterized by the presence of increased numbers of activated macrophages. These macrophages generate several inflammatory cell chemoattractants, by which neutrophil migrate from vascular compartment to the alveolar space. Recruited neutrophils secrete toxic oxygen radicals or proteolytic enzymes and induce inflammatory response. Continuing inflammatory response results in alteration of the pulmonary structure and irreversible fibrosis. Recently, a polypeptide with specific neutrophil chemotactic activity, interleukin-8(IL-8), has been cloned and isolated from a number of cells including : monocytes, macrophages and fibroblasts. IL-1 and/or TNF-${\alpha}$ preceded for the synthesis of IL-8, and we already observed high level of IL-1 and TNF-${\alpha}$ in the pneumoconioses. So we hypothesized that IL-8 may be a central role in the pathogenesis of pneumoconiosis. In order to evaluate the clinical utility of IL-8 as a biomarker in the early diagnosis of pneumoconiosis, we investigated the increase of IL-8 in the pneumoconiotic patient and the correlation between IL-8 level and progression of pneumoconiosis. Method : We measured IL-8 in the serum of 48 patients with pneumoconiosis and 16 persons without dust exposure history as a control group. Pneumoconiotic cases were divided into 3 groups according to ILO Classification : suspicious group(n=16), small opacity group(n=16) and large opacity group(n=16). IL-8 was measured by a sandwich enzytne immunoassay technique. All data were expressed as the $mean{\pm}standard$ deviation. Results: 1) The mean value of age was higher in the small opacity and large opacity group than comparison group, but smoking history was even. Duration of dust exposure was not different among 3 pneumoconiosis groups. 2) IL-8 level was $70.50{\pm}53.63pg/m{\ell}$ in the suspicious group, $107.50{\pm}45.88pg/m{\ell}$ in the small opacity group, $132.50{\pm}73.47pg/m{\ell}$ in the large opacity group and $17.85{\pm}33.85pg/m{\ell}$ in the comparison group. IL-8 concentration in all pneumoconiosis group was significant higher than that in the comparison group(p<0.001). 3) IL-8 level tended to increase with the progression of pneumoconiosis. Multiple comparison test using Anova/Scheffe analysis showed a significant difference between suspicious group and large opacity group(p<0.05). 4) The level of IL-8 was correlated with the progression of pneumoconiosis(r=0.4199, p<0.05). Conclusion : IL-8 is thought to be a good biomarker for the early diagnosis of pneumoconiosis.

  • PDF