• Title/Summary/Keyword: CAT protein

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Anti-oxidative and Anti-hyperglycemia Effects of Dung Beetle Extracts on the High Fat Diet SD Rats (고지방식이로 유발한 흰쥐에서 쇠똥구리 추출물의 항산화 효과 및 혈당강하에 미치는 영향)

  • Kim, Ha Jeong;Kim, Ban Ji;Ahn, Mi Young
    • Journal of Life Science
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    • v.26 no.7
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    • pp.772-781
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    • 2016
  • Dung beetle (Catharsius molossus, CA) is a well-known group of insects thanks to their exploitation of animal feces, a behavioral trait with a global impact on earth′s ecosystems. This study was conducted to investigate the effect of CA extract on a high-fat diet in SD rats. Male rats were divided into 5 groups. Animals were fed on a high-fat diet for seven weeks before and dung beetle extract for a month during the administration. Weight gain was decreased in ethanol extract from CA group. Administration of CA extract reduced the organ weight of testis and kidney, and adipose tissue weight. Lipid oxidative stress was evaluated measuring malondialdehyde level in liver. There were no significant differences in groups. Protein oxidative stress was evaluated measuring protein carbonyl content in blood. The protein carbonyl in blood was significantly decreased in ethanol and acetone extracted dung beetle groups (p<0.05). Meanwhile, the protein carbonyl in hepatocyte was not significant among the groups. Fibronectin and laminin by using D-HUVEC cell in vitro were measured by ELISA assay. There was significance in CA extract. The level of IL-10, IL-1β, VEGF, eNOS was evaluated by ELISA. There was significance in IL-10 compared to control (p<0.05). SOD and GPx tended to increase by CA extract. Furthermore, CAT was increased significantly by CA extract (p<0.05). After administration of CA extracts the composition of saturated fatty acid in adipose tissue tend to decrease, while unsaturated fatty acid increases. In conclusion, dung beetle had anti-hyperglycemia effects of oxidative stress and antioxidant activity.

Regulation of the Contraction Induced by Emptying of Intracellular $Ca^{2+}$ Stores in Cat Gastric Smooth Muscle

  • Baek, Hye-Jung;Sim, Sang-Soo;Rhie, Duck-Joo;Yoon, Shin-Hee;Hahn, Sang-June;Jo, Yang-Hyeok;Kim, Myung-Suk
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.2
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    • pp.113-120
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    • 2000
  • To investigate the mechanism of smooth muscle contraction induced by emptying of intracellular $Ca^{2+}$ stores, we measured isometric contraction and $^{45}Ca^{2+}$ influx. $CaCl_2$ increased $Ca^{2+}$ store emptying- induced contraction in dose-dependent manner, but phospholipase C activity was not affected by the $Ca^{2+}$ store emptying-induced contraction. The contraction was inhibited by voltage-dependent $Ca^{2+}$ channel antagonists dose dependently, but not by TMB-8 (intracellular $Ca^{2+}$ release blocker). Both PKC inhibitors (H-7 and staurosporine) and tyrosine kinase inhibitors (genistein and methyl 2,5-dihydroxycinnamic acid) significantly inhibited the contraction, but calmodulin antagonists (W-7 and trifluoperazine) had no inhibitory effect on the contraction. The combined inhibitory effects of protein kinase inhibitors, H-7 and genistein, together with verapamil were greater than that of each one alone. In $Ca^{2+}$ store-emptied condition, $^{45}Ca^{2+}$ influx was significantly inhibited by verapamil, H-7 or genistein but not by trifluoperazine. However combined inhibitory effects of protein kinase inhibitors, H-7 and genistein, together with verapamil were not observed. Therefore, this kinase pathway may modulate the sensitivity of contractile protein. These results suggest that contraction induced by emptying of intracellular $Ca^{2+}$ stores was mediated by influx of extracellular $Ca^{2+}$ through voltage-dependent $Ca^{2+}$ channel, also protein kinase C and/or tyrosine kinase pathway modulates the $Ca^{2+}$ sensitivity of contractile protein.

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A Case of Nasal Cryptococcosis in a Domestic Shorthair Cat (코리안 쇼트헤어 고양이에서 발생한 크립토코쿠스 감염증 의심 1증례)

  • Lee, Jin Soo;Kim, Hyun Wook;Choi, Ul Soo
    • Journal of Veterinary Clinics
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    • v.30 no.2
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    • pp.115-118
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    • 2013
  • An 8-year-old spayed female domestic shorthair cat was presented with a chief complaint of chronic nasal discharge and dyspnea. Physical examination revealed pyohemorrhagic nasal discharge, inspiratory dyspnea and stertor, and an enlarged right mandibular lymph node. Abnormalities of blood works and serum chemistry included mildly increased hematocrit, and globulin concentration. Serologic tests for FeLV and FIV, and a panel of polymerase chain reaction tests for Chlamydophila felis, Feline Calicivirus, Herpesvirus, Bordetella, Mycoplasma felis, and H1N1 influenza was all negative. Only radiographic finding showed increasing soft tissue density in the right nasal cavity and computed tomography disclosed soft tissue/fluid opacification in the right nasal cavity, paranasal sinus, and pharyinx along with slight deviation to the right of the osseous nasal septum. Focal lysis of ventral nasal septum was also suspected in CT scan. Cytological evaluation of fine needle aspirate smears of the enlarged mandibular lymph nodes revealed numerous fungal yeasts having variably thick capsule both extracellularly and intracellularly with low numbers of macrophages. Some yeasts showed narrow based budding, which was a consistent finding with Cryptococcus organisms. Serum protein electrophoresis was a polyclonal consistent with chronic infection and serum was submitted for a fungal serology panel test. In serologic tests Cryptococcus antigen titer was 1 : 32,768. In vitro culture was unsuccessful. Treatment was initiated with administration of fluconazole, clindamycin, and tocopherol. Clinical signs resolved within 3 days after the initial treatment. The cat was discharged and scheduled for periodic evaluation and continued therapy, but was lost to follow-up thereafter.

Effects of Sea Tangle (Laminaria japonica) Extract and Fucoidan Drinks on Oxygen Radicals and Their Scavenger Enzymes in Stressed Mouse (스트레스 부하 마우스의 활성산소 및 제거효소에 미치는 다시마(Laminaria japonica)와 후코이단 음료의 영향)

  • CHOI Jin-Ho;KIM Dae-Ik;PARK Soo-Hyun;KIM Dong-Woo;KIM Chang Mok;KOO Jae Geun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.6
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    • pp.764-769
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    • 1999
  • This study was designed to investigate the effects of sea tangle (Laminaria japonica) extract (Dasi-Ex group: dry base $4.0\%$) and fucoidan-added (Fuco-I, II, III group: fucoidan of $1,0\%,\;2.0\%,\;3.0\%$ added to Dasi-Ex) drinks on the formation of oxygen radicals and scavenger enzyme activities of stressed mice. ICR male mice (20 $\pm$2 g) were fed experimental diets and these drinks instead of water for 18 days including 4 days of sociopsychological stress. Dasi-Ex and Fuco-I, II and III groups resulted in a marked decreases $20\~40\%$ in basal oxygen radical (BOR) formation, and $15\~25\%$ in induced oxygen radical (IOR) formation compared with control group. Hydroxyl radical formations were significantly inhibited about $10\%$ in Dasi-Ex group, while remarkably inhibited $30\~40\%$ in Fuco-I, II and III groups. lipid peroxide (ISO) levels in Dasi-Ex group were not significantly different from those of control group, tut Fuco-I, II and III groups resulted in a significant decreases about $10\%$ in LPO levels compared with control group, Dasi-Ex, Fuco-I, II and III groups resulted in a marked decreases ($31\%,\;36\%,\;39\%$ and $42\%$, respectively) in oxidized protein levels through production of carbonyl group. Significant differences in nitric oxide (NO) levels in Dasi-Ex group were not obtained, but NO levels were slightly inhibited about $7\%$ in Fuco-I and II groups and $20\%$ in Fuco-III group compared with control group. Significant differences in superoxide dismutase (SOD) and catalase (CAT) activities in Dasi-Ex and Fuco-I groups were not obtained, but Fuco-II and III groups resulted in a significant increases $25\~40\%$ in SOD activities, and about $10\%$ in CAT activities compared with control group. These results suggest that the sociopsychological stress and aging process could be effectively inhibited by biological activity of sea tangle and fucoidan components.

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Functional Analysis and Selection of Second-site Revertant of Escherichia coli 16S rRNA of C770G (Escherichia coli 16S rRNA 상의 770 위치에 염기치환을 가진 변이체 리보솜의 단백질 합성 능력을 회복시키는 이차복귀돌연변이체의 발췌)

  • Ha, Hye-Jeong;Ryou, Sang-Mi;Lee, Kang-Seok;Jeon, Che-Ok
    • Microbiology and Biotechnology Letters
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    • v.39 no.1
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    • pp.93-96
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    • 2011
  • It has been shown that a nucleotide substitution at position 770 in Escherichia coli 16S rRNA, which is implicated in forming the evolutionary conserved B2c intersubunit bridge, has a detrimental effect on ribosome function. In order to isolate second-site revertants that complement ribosomes containing C770G, we performed a random mutagenesis of the 16S rRNA gene and selected clones that could produce more CAT protein translated by specialized ribosome. One of the clones contained two nucleotide substitutions at positions 569 and 904 (C569G and U904C) and these mutations partially complemented the loss of protein-synthesis ability caused by C770G. Further studies using the isolated revertant will provide information about which part of 16S rRNA is interacting with C770 and the consequence of the structure formed by these interactions in the process of protein synthesis.

Study on the expression and detection of the p53 mutation in Korean colon cancer cell lines (한국인의 대장암 세포주에서 p53 돌연변이의 발견과 발현에 관한 연구)

  • Jung, Ji-Yeon;Oh, Sang-Jin
    • IMMUNE NETWORK
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    • v.1 no.2
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    • pp.151-161
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    • 2001
  • Background: Inactivation in p53 tumor suppressor gene through a point mutation and deletion is one of the most frequent genetic changes found in human cancer, with 50% of an incidence. This high rate of mutation mostly suggests that the gene plays a central role in the development of cancer and the mutations detected so far were found in exons 5 to 8. Mutation of p53 locus produced accumulation of abnormal p53 protein, and negative regulation of cell proliferation and transcriptional activation as a suppressor of transformation were lost. In addition, inhibition of its normal cellular function of wild-type by mutant is an important step in tumorigenesis. Method: 4 colon cancer cell lines (SNU C1, C2A, C4, C5) were examined for mutation in exons 5 to 8 of the p53 tumor suppressor gene by PCR-SSCP analysis and expression pattern by western blotting and immunoprecipitation. p53-mediated transactivation ability were examined by CAT assay and base substitution of p53 in SNU C2A cell were detected by DNA sequencing. Results: 1) SNU C2A cell and SNU C5 cell were detected mobility shifts each in exon 5 and exon 7 of p53 gene by the PCR-SSCP method, implicating being of p53 mutation. 2) 3 colon cancer cell lines (SNU C1, SNU C2A, SNU C5) expressed wild type and mutant type p53 protein. 3) In northern blot experiment, SNU C2A and SNU C5 cell expressed high level of p53 mRNA. 4) Results of p53-mediated transactivation in colon cancer cell lines by CAT assay represented only SNU C2A cell has transcriptional activity. 5) DNA sequencing in SNU C2A cell showed missense mutation in codon 179 of one allele, histidine to arginine and wild type p53 in the other allele. Conclusion: Colon cancer cell lines showed correlation with mutation in p53 gene and accumulation of abnormal p53 protein. Colon cancer cell SNU C2A retained p53-mediated transactivation as heterozygous p53 with one mutant allele in 179 codon and the other wild-type allele.

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Mycobacterium tuberculosis ESAT6 and CPF10 Induce Adenosine Deaminase 2 mRNA Expression in Monocyte-Derived Macrophages

  • Bae, Mi Jung;Ryu, Suyeon;Kim, Ha-Jeong;Cha, Seung Ick;Kim, Chang Ho;Lee, Jaehee
    • Tuberculosis and Respiratory Diseases
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    • v.80 no.1
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    • pp.77-82
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    • 2017
  • Background: Delayed hypersensitivity plays a large role in the pathogenesis of tuberculous pleural effusion (TPE). Macrophages infected with live Mycobacterium tuberculosis (MTB) increase the levels of adenosine deaminase2 (ADA2) in the pleural fluid of TPE patients. However, it is as yet unclear whether ADA2 can be produced by macrophages when challenged with MTB antigens alone. This study therefore evaluated the levels of ADA2 mRNA expression, using monocyte-derived macrophages (MDMs) stimulated with MTB antigens. Methods: Purified monocytes from the peripheral blood mononuclear cells of healthy volunteers were differentiated into macrophages using granulocyte-macrophage colony-stimulating factor (GM-CSF) or macrophage colony-stimulating factor (M-CSF). The MDMs were stimulated with early secretory antigenic target protein 6 (ESAT6) and culture filtrate protein 10 (CFP10). The mRNA expression levels for the cat eye syndrome chromosome region, candidate 1 (CECR1) gene encoding ADA2 were then measured. Results: CECR1 mRNA expression levels were significantly higher in MDMs stimulated with ESAT6 and CFP10, than in the unstimulated MDMs. When stimulated with ESAT6, M-CSF-treated MDMs showed more pronounced CECR1 mRNA expression than GM-CSF-treated MDMs. Interferon-${\gamma}$ decreased the ESAT6- and CFP10-induced CECR1 mRNA expression in MDMs. CECR1 mRNA expression levels were positively correlated with mRNA expression of tumor necrosis factor ${\alpha}$ and interleukin 10, respectively. Conclusion: ADA2 mRNA expression increased when MDMs were stimulated with MTB antigens alone. This partly indicates that pleural fluid ADA levels could increase in patients with culture-negative TPE. Our results may be helpful in improving the understanding of TPE pathogenesis.

Changes in the Myocardial Antioxidant Enzyme System by Post-Ischemic Reperfusion During Corontory Artery Bypass Operations (관상동맥우회술시 심근허혈후 재관류에 의한 활성산소 방어효소계의 변화)

  • 김응중;김기봉
    • Journal of Chest Surgery
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    • v.29 no.8
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    • pp.850-860
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    • 1996
  • Oxygen free radicals and their metabolites have been implicated as possible causes of reperrusion injury In animal models. Their role in the clinical setting is still controversial. The aim of this study was to evaluate the degree of tissue damage, oxidative stress. and changes in the antioxidant enzyme system in patients undergoing cor nary artery bypass graft operations(CABG) with myocardial protection by cold blood cardioplegia. In patients undergoing CABG(n:10). the levels of lactate dehydrogenate(LDH), creatine phosphokinase MB fraction(CK-MB), and malondialdehyde(M DA) were measured In the coronary sinus effluent before aortic cross clamping and 20 minutes after reperfusion. At the same time, the myocardial tissue activities of superoxide dismutase(SOD). catalase(CAT), glutathione peroxiddse(GSHPX), glutathione reductase (GSSGRd), and glucose 6-phosphate dehydrogenate(GfPDH ) were determined in the right atrial auricle excised before aortic cross clamping and in the left atrial auricle excised 20 minutes after reperfuslon. The levels of increased significantly after reperrusion(p< U.05). There were no significant changes in CAT and CfPDH levels. Western blot analysis was performed to study the induction of antioxidant enzyme and demonstrated increased amount of Cu,Zn-SOD.

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Anti-inflammatory Efficacy of HK Shiitake Mushroom Mycelium in LPS-treated RAW 264.7 Cells Through Down-regulation of NF-κB Activation (LPS로 활성화한 RAW 264.7 세포에서 HK표고버섯균사체의 NF-κB 활성 억제를 통한 항염증 효과)

  • Song, Chae Yeong;Oh, Tae Woo;Kim, Hoon Hwan;Lee, Yu Bin;Kim, Jeong Ok;Kim, Gon Sup;Ha, Yeong Lae
    • Journal of Life Science
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    • v.32 no.7
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    • pp.491-500
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    • 2022
  • HK shiitake mushroom mycelium (HKSMM), containing 14% β-glucan, is a health functional food ingredient individually approved by the Korea Ministry of Food and Drug Safety for liver health. The anti-inflammatory effect of a 50% aqueous ethanol extract of HKSMM (designated HKSMM50) was studied in RAW 264.7 macrophage cells treated with lipopolysaccharide (LPS). An active hexose correlated compound (AHCC) was used as a positive control. LPS-activated RAW 264.7 cells were treated with HKSMM50 and AHCC (0, 20, 100, 500 ㎍/ml) and cultured for 24 hr. Inflammation-related elements in the supernatant were measured using enzyme-linked immunosorbent assay (ELISA) kits, and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins in the cells was analyzed by Western blotting. The HKSMM50 lowered iNOS and COX-2 protein expressions, and nuclear factor-kappa B (NF-κB), nitric oxide (NO) and prostaglandin E2 (PGE2) contents in a concentration-dependent manner as compared to LPS treatment. Similarly, the HKSMM50 lowered the content of pro-inflammatory cytokines interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) and interleukin-6 (IL-6) contents and increased the activity of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). The efficacy of the AHCC treatment was similar to that of the HKSSM50 treatments. These results indicate that HKSMM50 showed an anti-inflammatory effect in LPS-treated RAW 264.7 cells by down-regulation of NF-κB signaling and suggest that HKSMM could be used as a health functional food ingredient to help improve immune function.

Antioxidant Enzyme Activity and Antimicrobial Activity of Isatis tinctoria Extract (대청 추출물의 항산화 효소 활성 및 항균 효과)

  • Heo, Buk-Gu;Park, Yun-Jum;Lee, Seung-Jin;Kim, Kwan-Su;Cho, Ja-Yong;Boo, Hee-Ock
    • Korean Journal of Plant Resources
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    • v.25 no.5
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    • pp.543-549
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    • 2012
  • To gather the basic data for increasing the utilization of Isatis tinctoria, we examined the effects of both antioxidative enzyme activity and antimicrobial activity from the extract of Isatis tinctoria. Ascorbate Peroxidase activities reveal that there is an decrease in order; ethanol extract from its stem (1601.7 Unit/mg protein), methanol extract from its leaf (1133.7 Unit/mg protein) and distilled water extract from its leaf (524.3 Unit/mg protein). Catalase activities reveal that there is an decrease in order; ethanol extract from its flower petal (177.1 Unit/mg protein), methanol extract from its leaf (120.8 Unit/mg protein) and distilled water extract from its flower petal (55.4 Unit/mg protein). Peroxidase activities reveal that there is an decrease in order; ethanol extract from its flower petal (27.1 Unit/mg protein), methanol extract from its flower petal (14.6 Unit/mg protein) and distilled water extract from its stem (10.4 Unit/mg protein). Superoxide dismutase activities reveal that there is an increase in order; distilled water extract from its root (90.8%), methanol extract from its flower petal (80.1%) and ethanol extract from its root (75.5%). Its flower extract showed a antimicrobial activity only against Vibrio parahaemolyticus, its root extract had only against Staphylococcus aureus, and its stem extract had against Bacillus subtilis, Escherichia coli and Staphylococcus aureus, regardless of solvents. Especially, distilled water extract from its leaf showed a high antimicrobial activity against both Bacillus subtilis and Escherichia coli and inhibition diameters against those were 30.0 and 24.0 mm, respectively.