• Title/Summary/Keyword: Proteinase

Search Result 389, Processing Time 0.023 seconds

Alu-Derived Alternative Splicing Events Specific to Macaca Lineages in CTSF Gene

  • Lee, Ja-Rang;Park, Sang-Je;Kim, Young-Hyun;Choe, Se-Hee;Cho, Hyeon-Mu;Lee, Sang-Rae;Kim, Sun-Uk;Kim, Ji-Su;Sim, Bo-Woong;Song, Bong-Seok;Jeong, Kang-Jin;Lee, Youngjeon;Jin, Yeung Bae;Kang, Philyong;Huh, Jae-Won;Chan, Kyu-Tae
    • Molecules and Cells
    • /
    • v.40 no.2
    • /
    • pp.100-108
    • /
    • 2017
  • Cathepsin F, which is encoded by CTSF, is a cysteine proteinase ubiquitously expressed in several tissues. In a previous study, novel transcripts of the CTSF gene were identified in the crab-eating monkey deriving from the integration of an Alu element-AluYRa1. The occurrence of AluYRa1-derived alternative transcripts and the mechanism of exonization events in the CTSF gene of human, rhesus monkey, and crabeating monkey were investigated using PCR and reverse transcription PCR on the genomic DNA and cDNA isolated from several tissues. Results demonstrated that AluYRa1 was only integrated into the genome of Macaca species and this lineage-specific integration led to exonization events by producing a conserved 3' splice site. Six transcript variants (V1-V6) were generated by alternative splicing (AS) events, including intron retention and alternative 5' splice sites in the 5' and 3' flanking regions of CTSF_AluYRa1. Among them, V3-V5 transcripts were ubiquitously expressed in all tissues of rhesus monkey and crab-eating monkey, whereas AluYRa1-exonized V1 was dominantly expressed in the testis of the crab-eating monkey, and V2 was only expressed in the testis of the two monkeys. These five transcript variants also had different amino acid sequences in the C-terminal region of CTSF, as compared to reference sequences. Thus, species-specific Alu-derived exonization by lineage-specific integration of Alu elements and AS events seems to have played an important role during primate evolution by producing transcript variants and gene diversification.

Anti-skin-aging Effect of Mori Folium through decreased Advanced glycation end product (AGEs) (최종당화산물 억제를 통한 상엽(桑葉)의 항피부노화 효과)

  • Lee, AhReum;Kim, SooHyun;Kim, SuJi;Kim, KyeongJo;Kwon, Ojun;Choi, JoonYoung;Roh, Seong-Soo
    • The Korea Journal of Herbology
    • /
    • v.32 no.5
    • /
    • pp.7-12
    • /
    • 2017
  • Objectives : Recently, numerous studies reported that excessive generation of advanced glycation end products (AGEs) stimulated expression of skin wrinkle related proteins. This study aimed to evaluate inhibits skin wrinkle formation effect of Mori Folium (MF) through decreased AGEs. Methods : To evaluate the skin wrinkle inhibition effect of MF, SD-rats were divided into three groups; normal rats (Nor), AGEs-induced rats (Con), AGEs-induce rats treated with MF at dose of 100mg/kg body weight (MF). To induced AGEs, streptozotocin (50mg/kg) was injected intraperitoneally, and after 3 days, 100mM methyl glyoxal was administered orally for 3 weeks. After the experiment, the animal's dorsal skin tissues and serum were separated and tested. Results : The oral administration of MF suppressed the AGEs level in serum. Also, the AGEs in skin tissues was significantly reduced through treatment of MF compared with control group. Moreover, the expressions of AGEs related proteins such as polyclonal anti-$N^e$-(carboxymethyl) lysine (CML), anti-$N^e$-(carboxyethyl)lysine (CEL), AGE receptors (RAGE) were reduced in MF group compared with the control group in kidney and skin tissues. The matrix metallo proteinase-1 (MMP-1) reduced by MF treatment with the result that collagen type 1 alpha 2 (COL1A2) was improved that reduced by accumulation of AGEs. Conclusion : The evidence of this study indicate that oral administration of MF reduces the levels of AGEs in serum, skin, and kidney tissues. In conclusion, MF inhibit skin wrinkle formation, suggesting the potential of anti-wrinkle material.

Antioxidant Properties of Peptides Extracted from Tenebrio molitor Larvae (갈색거저리 유충에서 추출한 펩타이드의 항산화 특성)

  • Sam Woong Kim;Sang Wan Gal;Won-Jae Chi;Woo Young Bang;So Jeong Park;Tae Wan Kim;Kyu Ho Bang
    • Journal of Life Science
    • /
    • v.33 no.5
    • /
    • pp.383-390
    • /
    • 2023
  • The goal of this study was to identify new bioactive peptides in extracts derived from Tenebrio molitor (T. molitor) larvae for the development of functional foods. After extraction from freeze-dried T. molitor larvae with various solvents on time course, the extracts showed the highest 2,2,1-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity at 5 and 10 hr per total protein and solid contents, respectively. When the water extract was fractionated, a high methanol concentration led to a reduced level of high-molecular-weight proteins in the centrifugal supernatant, whereas increased DPPH activity in the supernatants suggests low-molecular-weight peptides may mediate antioxidant activity in the supernatant. Most of the organic solvent partitions, excluding butanol, showed similar activities in the water phases, and the organic solvent partition fraction exhibited a 28~44% decrease in activity following heat treatment, implying that some components in the fraction become unstable in the presence of heat. The addition of proteinase K to the water extract increased DPPH activity by 10~20%, suggesting that peptides, when released from total proteins, partially increase antioxidant activity. Therefore, we suggest that the antioxidants in T. molitor larval extracts make them a potential source of functional animal food.

Expression of Matrix Metalloproteinase-9 and Tissue Inhibitor of Metalloproteinase-1 after Administration of Endotoxin in Diabetic Rats (내독소로 자극된 당뇨 쥐에서 단백분해효소와 그 억제제 발현)

  • Seo, Ki Hyun;Choi, Jae Sung;Na, Joo Ok;Uh, Soo Taek;Kim, Yong Hoon;Park, Choon Sik
    • Tuberculosis and Respiratory Diseases
    • /
    • v.61 no.3
    • /
    • pp.256-264
    • /
    • 2006
  • Background: An acute lung injury(ALI) is characterized by the recruitment, activation, and apoptosis of inflammatory cells, numerous products released by inflammatory cells such as reactive oxygen species, inflammatory mediators, and a variety of proteolytic enzymes. It was reported that bacterial infections in diabetics showed impaired PMN functions such as reduced PMN respiratory burst and decreased microbicidal activity in inflamed tissue. However, the effect of the proteinase - inhibitor (MMP-9 vs TIMP-1) in ALI in diabetics is unclear. This study evaluated the differences in the expression of MMP-9 and TIMP-1 after the stimulation of endotoxin in a rat model. Methods: Six-week-old male Sprague-Dawley rats were classified into normal, DM, LPS and DM+LPS groups. The peripheral blood, BAL fluids, and lung tissues were obtained from individual rats. The MMP-9 activity was measured by gelatin zymography and the TIMP-1 level was measured by Western blotting. Results: The total BAL cells of the DM-LPS groups were significantly lower than the LPS groups (p < 0.01). The MMP-9 activities in the serum were higher in the DM+LPS groups than in the other groups. The MMP-9 activities in the BAL fluids were significantly higher in the DM+LPS group than in the normal and diabetic rats (p < 0.05). TIMP-1 expressions in the BAL fluids were significantly lower in the DM+LPS group than other groups (p < 0.05). The ratio between MMP-9 and TIMP-1 in the BAL fluids was significantly higher in the DM+LPS groups (p < 0.05). Conclusion: In ALI in diabetics the higher MMP-9 activity and lower TIMP-1 level are believed to prolonged and intensify the course of inflammation.

Biosynthesis of the extracellular enzymes in de novo during the differentiation of Aspergillus niger (검정곰팡이의 형태분화에 따른 세포외성효소의 신생적생합성에 관한 연구)

  • Kim, Jong-Hyup
    • The Korean Journal of Mycology
    • /
    • v.6 no.2
    • /
    • pp.1-10
    • /
    • 1978
  • In de novo biosynthesis of the extracellulor enzymes-proteinsaes, alpha and gluc-amylases during the synchronized differentiation of Aspergillus niger in submerged culture and surface liquid culture were investigated. Gluc-amylase was synthesized in the stage of presporulation in which phialide formation is involved. Proteinase was synthesized both in the stages of conidiophore formation and presporulation. Alpha-amylase was synthesized during presporulation and sporulation stages, the activity of enzyme lasted for seven days on surface liquid culture. It seemed that the synthesis was occured in de novo partly repressed by the catabolite, and its nature was found to be constitutive since it is produced in non-starch medium. Polyacrylamide gel electrophoresis have shown that presporulating and sporulating body produced diverse types of the proteins whereas the earlier stages of vegetative body showed simpler profiles. The uptake of C-14 uracil into RNA and C-14 glutamate into protein were shown to be vigorous in presporulating body rather than those in sporulating body. Coincidence of alpha-amylase biosynthesis in de novo and sporulation may be significant in the study of differentiation in which gene expression is involved.

  • PDF

Differentiation of Entomoeba histolyticn and Entcmoeba dispor in cyst-passers by immunoblot (면역이적법을 이용한 아질아메바와 동형아메바의 감별진단)

  • 이미정;홍성태
    • Parasites, Hosts and Diseases
    • /
    • v.34 no.4
    • /
    • pp.247-254
    • /
    • 1996
  • Differentiation of invasive strains of Entamoebn histolytica according to their pathogenicity has been a topic of long debate, but now the pathogenic species only is regarded as E. histolytica while the non-pathogenic species is E. dispar. The present study applied immunoblot to differentiale infections of the two species among microscopically- detected cyst-passers in Korea. The crude extract of 5. histolyticn separated in 5-20% gradient gels, revealed many fractions of 94. 81. 71, 50. 44, 38.5. 37.5, 29, 19. and 18 kDa when the cysteine proteinase inhibitor. E64, was supplemented. The serum IgG antibody of 3 proven E. histolytirc cases reacted loth the antigenic fractions of 117. 110. 99.68,66,60.54.52, 46. and 45 kDa. Sera of PCR confirmed 3 cases of E. disper reacted only to the 117 kDa fraction or the E. histolytica crude extract which was regarded as non specific. To the antitigen of monoxenic E. dispar. sera or E. dispar and E. histolytica cases showed the same immunoblot reactions. The serum IgG antibody reacted with several antigenic fractions of both E. histolytica and E. dispar. but IgM and IgE antibodies showed no reaction to either antigen. Sera of 24 symptomless amebic cyst-passers were screened with the E. histolytica alltigen; two were found to be infected by E. histolytica and 22 were by E. dispar. The present findings suggest that in Korea most of asymptomatic cyst passers of E. histolytica are carriers of E. dispar. Immunoblot using E. histolytica antigen is a good technique for the differentiation of E. histolytica and E. dispar infections.

  • PDF

A comparative study on denture cleansers for food stain removal on the denture of elders in domiciliary care (재가복지 노인의치 세정제의 음식물 세정효과 비교연구)

  • Lee, Nam-Soon
    • Journal of the Korea Academia-Industrial cooperation Society
    • /
    • v.19 no.1
    • /
    • pp.331-336
    • /
    • 2018
  • Oral health is an essential requirement for maintaining and promoting health and quality of life while aging. It is known that there is a deep connection with hygienic management of dentures. However, despite the emphasis on the importance of clean management of dentures, there is a lack of quantitative research on denture-related detergents. Therefore, the purpose of this study is to evaluate food stain removal performance of three denture cleansers containing proteolytic enzymes. In addition, this study provides basic data on self-cleaning methods using detergents and suggests ways to improve effective treatment for oral health of elders in domiciliary care. In order to obtain quantitative data concerning food stain removal performance of each proteinase denture cleaner, the test evaluation method proposed by the Korean Consumer Agency, ISO 20795, was used. A total of 18 specimens were discolored for 48 hours in water dissolved with turmeric, and then washed with each denture cleaner for 24 hours. The cleaned denture specimens were measured quantitatively using a spectrophotometer to measure the color values of the denture specimens before the discoloration as well as the color values of the discolored specimens after cleaning with the denture cleanser. The experimental values were analyzed through one-way ANOVA with post-hoc Tukey's test, and statistically significant differences were found among the experimental groups. It was found that C had statistically significant stain removal performance compared with A and B(p<.05). All experiments were conducted for a total of 21 days, from July 3- 23, 2017. This study confirms that denture cleansers is a beneficial self-denture management method for domiciliary seniors. Therefore, it is necessary to extend accessibility for denture cleansers in order to to optimize domiciliary care for elderly oral hygiene and health.

Thromboelastographic Analysis of the Coagulation System During Cardiopulmonary Bypass -Analysis of the Effect of Low-Dose Aprotinin (심폐바이패스시 혈액응고체계 변화의 혈전탄성검사 분석 - 단일 저용량 아프로티닌 투여 효과 분석 -)

  • 김관민;박계현
    • Journal of Chest Surgery
    • /
    • v.30 no.7
    • /
    • pp.677-685
    • /
    • 1997
  • Thromboelastography(TEG) is the unique measure that gives rapid information about the whole clotting process. Simplifying the diagnosis of coagulopathy during operations, TEG can provide an adequate therapy for postoperative bleeding. Remarkable improvement in hemostasis after cardiopulmonary bypass(CPB) has been achieved by the treatment with proteinase inhibitor aprotinin, but the hemostatic mechanism of aprotinin during CPB is still unclear. This study was designed to evaluate the effects of aprotinin on coagulation system during CPB by using TEG. Forty patients who underwent CPB were divided into two groups: aprotinin(2u 106 kallikrein inhibition units, as a single dose into the cardiopulmonary bypass priming solution) treatment group(male 14, female 8, mean age=50.Byears) and no aprotinin treatment(control) group(male 10, female 8, mean age=53.4 years). TEG, activated clotting time, prothrombin time, activated partial thromboplastin time, platelet counts, fibrinogen an (ibrinogen degradation product(FDP) concentrations were checked before and after CPB(30 minutes after neutralization of heparin effect by protamine sulfate). There was no significant difference in other conventional coagulation tests of two groups except postcardiopulmonary bypass FDP concentration in control group, which was significantly increased compared to that in aprotinin group(p<0.05). In TEG variables of both groups, clot formation time(K) and alpha $angle(\alpha^{\circ})$ were significantly increased and decreased, respectively, after CPB(p<0.05), but fibrinolytic index(LYS60) was not changed during CPB. In aprotinin group, reaction time(R) was decreased significantly after CPB(p<0.05) but maximum amplitude(MA) was not changed(p>0.05). On the contrary, R was not changed markedly but MA was decreased significantly in control group after CPB(p<0.05). This result shows that main change in coagulation system during CPB is not hyperfibrinolysis but cecrease in clot strength by platelet dys unction, and the main effect of aprotinin during cardiopulmonary bypass is the maintenance of clot strength to the pre-CPB level by the preservation of platelet function.

  • PDF

Urokinase Plasminogen Activator Receptor Gene Expression and Clinico-Pathologic Feature in Gastric Cancer Patients (위암 환자의 Urokinase Plasminogen Activator Receptor 유전자의 발현양상)

  • Kim Yong Gil;Lee Kyung Hee;Kim Min Kyung;Lee Jae Lyun;Hyun Myung Sue;Kim Sang Hun;Kim Hee Sun
    • Journal of Gastric Cancer
    • /
    • v.4 no.4
    • /
    • pp.207-212
    • /
    • 2004
  • Purpose: Invasion and metastasis in solid tumors require the action of tumor-associated proteases. The serine protease urokinase-type plasminogen (uPA) and receptor (uPAR) appear to have a major function in these processes. Expression of the uPAR is elevated in breast and colon carcinomas, and this is often associated with invasiveness and poor prognosis. The purpose of this study was to determine whether the expression of the uPAR gene correlates with clinico-pathological parameters in human gastric carcinomas. Materials and Methods: We examined the expression of uPAR mRNA by using northern blot analysis and RT-PCR in 35 gastric carcinomas and the surrounding normal mucosa. Macroscopic and histopathological tumor findings and survival rates were obtained from the patient records and from endoscopic, surgical, and pathological reports. Results: The expression of uPAR and was higher in most neoplasms than in the corresponding normal mucosal tissue. uPAR mRNA expression in tumors correlated well with lymph-node metastasis (P<0.02) and tumor stage (P<0.01). The survival rate of patients with tumors displaying high uPAR expression levels was significantly lower (P<0.04) than that of patients without uPAR expression, but IL-8 showed only the tendency of survival difference. Conclusion: These results suggest that uPAR may be an important prognostic factor in human gastric carcinomas.

  • PDF

QUANTITATIVE ANALYSIS OF TRANSFORMING GROWTH $FACTOR-{\beta}_1$ IN HUMAN FIBROBLASTS INDUCED WITH STAPHYLOCOCCUS ENTEROTOXIN B AND LIPOPOLYSACCHARIDE (Staphylococcus enterotoxin B와 lipopolysaccharide를 작용시킨 사람 섬유아 세포에서 생성된 Transforming Growth $Factor-{\beta}_1$의 정량적 분석)

  • Lee, Seong-Geun;Kim, Kwang-Hyuk;Kim, Uk-Kyu;Kim, Jong-Ryoul;Chung, In-Kyo;Yang, Dong-Kyu
    • Maxillofacial Plastic and Reconstructive Surgery
    • /
    • v.22 no.2
    • /
    • pp.123-132
    • /
    • 2000
  • $TGF-{\beta}_1$ is a potent chemotactic factor for inflammatory cells and fibroblasts. It also stimulates the celluar source and components of extracellular matrix and the production of proteinase inhibitors. Collectively, these biologic activities lead to the accumulation and stabilization of the nascent matrix, which is vital to infection control. The objective of this study is to investigate production of $TGF-{\beta}$ in vitro fibroblast culture in the presence of Staphylococcus enterotoxin B(SEB) and/or lipopolysaccharide(LPS) and to elucidate the role of $TGF-{\beta}_1$ which may be responsible for infection control. The fibroblasts were originated from gingiva and facial dermis in 26 year-old male patient. In the presence of LPS($0.01{\mu}g$, $0.1{\mu}g$, $1.0{\mu}g$), SEB($0.01{\mu}g$, $0.l{\mu}g$, $1.0{\mu}g$) respectively, $cells(5{\times}10^3ml)$ were cultivated in vitro. At 1, 3, and 5 days after incubation, cells were counted. Also, $cells(2.5{\times}10^5ml)$ were cultivated in EMEM with LPS(0.01, 0.1 and $1.0{\mu}g$), SEB(0.01, 0.1 and $1.0{\mu}g$) respectively and $LPS(0.1{\mu}g)$ and $SEB(0.1{\mu}g)$ in combination for 24, 48, and 72 hours respectively. Culture supernatants were harvested at 1, 2, and 3 days after incubation period and triplicate culture supernatants were pooled and $TGF-{\beta}_1$ was assayed in duplicate. The results were as follows. 1. In gingival fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell Proliferation occurred very significantly since 3 days after incubation, compared with the control and the production of $TGF-{\beta}_1$ occurred very significantly at 1 day after incubation, compared with the control. 2. In facial dermal fibroblast induced with SEB and LPS respectively or in combination, the suppression of cell proliferation occurred very significantly at 1 day after incubation, compared with the control. In SEB exposure, the production of $TGF-{\beta}_1$ was decreased very significantly at 1 day after incubation, compared with the control. However, in LPS, SEB and LPS exposure, the production of $TGF-{\beta}_1$ was increased very significantly at 1 day after incubation, compared with the control. In conclusion, the concentration of bacterial toxins and the incubation period correlated with cell proliferation and production of $TGF-{\beta}_1$ very significantly. The gingival and facial dermal fibroblasts have different phenotype each other The orchestrated understanding of fibroblast proliferation and $TGF-{\beta}_1$ production play an important part in host defense against the bacterial Infection and may prevent tissue necrosis such as necrotizing fasciitis and life-threatening syndrome such as multiple organ failure.

  • PDF