• Title/Summary/Keyword: oxidase test

Search Result 181, Processing Time 0.027 seconds

Expression and Significance of Hypoxia Inducible Factor-1α and Lysyl Oxidase in Non-small Cell Lung Cancer

  • Ping, Wei;Jiang, Wen-Yang;Chen, Wen-Shu;Sun, Wei;Fu, Xiang-Ning
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.6
    • /
    • pp.3613-3618
    • /
    • 2013
  • Object: To detect expression of hypoxia inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$) and lysyl oxidase (LOX) in non-small cell lung cancer (NSCLC) and explore their roles in prognosis. Methods: The mRNA levels of HIF-$1{\alpha}$ and LOX were investigated by real-time reverse-transcriptase polymerase chain reaction in 40 cases of tumour and paired normal tissues. In addition, protein expression of HIF-$1{\alpha}$ and LOX was examined by immunohistochemistry in 82 cases of tumour and 45 paired normal tissues. The relationship between HIF-$1{\alpha}$ or LOX and clinicopathologic characteristics, as well as the correlation between HIF-$1{\alpha}$ and LOX, were also examined. Kaplan-Meier survival curves and the log-rank test were used to analyze progression-free survival. Results: HIF-$1{\alpha}$ or LOX mRNA levels in tumor tissues was significantly higher than those in paired normal tissues (p<0.01). Positive HIF-$1{\alpha}$ or LOX protein expression in tumor tissues was noted in 46/82 (56.1%) and 49/82 (59.8%) of the cases, respectively, being significantly higher than those in paired normal tissues (p<0.05). There was significant correlation between the expression of HIF-$1{\alpha}$ or LOX and tumor size, lymph node metastasis and pathological stage (p<0.05). The expression of HIF-$1{\alpha}$ and LOX had a significant inverse impact on survival of patients with NSCLC. Conclusion: HIF-$1{\alpha}$ and LOX may play a pivotal role in the development of NSCLC, and may act in synergy to promote the progression of NSCLC.

An Assay of L-Ascorbic Acid with Crude Enzyme from Cucumber(Cucumis sativus L.) (오이(Cucumis sativus L.)에서 추출한 조효소 용액을 이용한 L-Ascorbic Acid의 정량분석)

  • Lee, Suk-Yeong;Kim, Hong-Seop;Go, Dae-Hui
    • The Korean Journal of Food And Nutrition
    • /
    • v.8 no.3
    • /
    • pp.159-164
    • /
    • 1995
  • The test possibility of L-ascorbic acid(AsA) assay by ascorbate oxidase(A.O) solution obtained from cucumber was estimated. The results obtained were summarized as follow. The difference of absorbance before and after oxidation of AsA by A.O solution was proportional to the AsA concentration. Iso- AsA compared to same concentration of AsA was 97% response. Therefore, the 0 specificity on Iso- AsA a was deficient. On the sucrose, glucose, fructose were coexisted from 5 times, 500 times of AsA concentration, none of them were affected by any level of AsA concentration. In the case of EDTA, it was not nearly affected that EDTA was coexisted 50 times of AsA concentration, but EDTA Inhibited considerably from 500 times of AsA concentration. The effect of citric acid on the AsA concentration was not in the same level of AsA, however, it was slightly inhibited from 5 times, 50 times of AsA concentration and 500 times of AsA inhibited remarkably As known L-AsA was catalycally autooxidated by Cu2+ and Fe2+. They were not nearly affected In the above 0.01M level on the Ash assay. However, Fe3+ ion was neither nearly affected in the 0.001M level nor In the above 0.01M level on the AsA assay.

  • PDF

Antioxidative Activity and Irritation Test of a Complex Herbal Bath Extracted from Korean Traditional Plants

  • Chang Zhiqiang;Kim Young-Hoan;Song Jae-Chan;Kim Nam-Woo;Lee In-Seon;Lee Sam-Pin;Kim Jong-Choon;Park Seung-Chun
    • Toxicological Research
    • /
    • v.21 no.1
    • /
    • pp.45-49
    • /
    • 2005
  • The purpose of this study is to evaluate the free radical scavenging ability and xanthine oxidase inhibitory activity of a complex herbal bath consisted of Artemisiae argyi folium, Angelicae sinensis radix, Ligustici wallichii radix and Angelicae tuhuo radix, and its potential irritation response were also tested for safety use in the rabbits. For antioxidative activity, the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of the complex herbal bath were examined at five different concentrations (0, 250, 500, 1000 and 2000 ㎍/ml). The concentration of the complex herbal bath required for scavenging DPPH free radical by 50% was 897.2 ㎍/ml. In the inhibition of xanthine oxidase (XO) activity, the concentration of the complex herbal bath required for 50% of inhibition was 221.4 ㎍/ml. In the skin irritation study in rabbit, all animals survived for the duration of the study and the examined skin exhibited no edema, erythema, and eschar formation. In the ocular irritation study in rabbit, after application of the sample to eyes, all of the eyes were normal. In summary, the complex herbal bath has potent antioxidant effects against the DPPH radical and XO and was considered to be a non-irritation bath for safety use.

Anti-oxidant and Anti-inflammation Activities of Prunus persica Flos (도화(桃花, Prunus persica Flos)의 항산화 및 항염증 활성)

  • Lee, Jin-Young;An, Bong-Jeun
    • Journal of Applied Biological Chemistry
    • /
    • v.53 no.3
    • /
    • pp.162-169
    • /
    • 2010
  • The solvent extracts of Prunus persica Flos were investigated for the activities of anti-oxidant and anti-inflammation to apply as a functional ingredient for cosmetic products. The electron donating ability of both ethanol (PPE) or acetone (PPA) extracts of P. persica Flos was above 90.0% at the concentration of 500ppm. The superoxide dismutase (SOD)-like activity of P. persica Flos extracts (PPE, PPA) were approximately 40.0% at 1,000 ppm. The xanthine oxidase inhibitory effect of P. persica Flos extracts (PPE, PPA) was approximately 30.0% at 1,000 ppm and equivalent to that of ascorbic acid. Hyaluronidase inhibition activity related to the anti-inflammation effect was 35.0% with the treatment of P. persica Flos extracts (PPW, PPE, PPA) at 1,000 ppm, respectively. In the experiment of anti-inflammation effect, P. persica Flos extracts (PPW, PPE, PPA) inhibited the generation of nitric oxide. In the antimicrobial activity test against the human skin-resident microflora such as Staphylococcus epidermidis and Propionibacterium acnes, a clear zone was identified from 4mg/disc in P. persica Flos (PPE) extract.

Cloning of Xanthine Oxidase Gene from Mouse Liver cDNA Library

  • Lee, Chu-Hee;Lee, Sang-Il;Nam, Doo-Hyun;Heo, Geun
    • Proceedings of the Korean Society of Applied Pharmacology
    • /
    • 1994.04a
    • /
    • pp.261-261
    • /
    • 1994
  • Bovine milk xanthine oxidase (E.C.1.1.3.22, XO) purchased from Sigma Chemical Co. had the three protein fragments below 150 kDa on 7.5% SDS-PAGE, which did not show enzyme activity. To remove these fragments, the enzyme preparation was further purified through Sephadex G-200 column chromatography. Two peaks exhibiting enzymatic activity were separated very closely to the void volume, which were revealed as two different enzyme forms, dimeric and monomeric, confirmed by activity staining on native PAGE. Anti sera-against each of the two enzyme forms were raised by subcutaneous injection at multiple sites on the back of rabbits during 4 weeks. On the immunodiffusion test, it was found that both of the antisera of the two forms could react with each other, which implied that their epitopes were identical In the Western blot analysis of mouse liver cytosol fraction, it was found that rabbit anti-XO antibody bound well with the protein band of monomeric mouse liver XO of about 150kDa. Based on this result, mouse liver cDNA 1 ibrary was screened by in situ hybridizat ion wi th rabbi t anti -XO antibody as probe. Through the immunological screening, recombinant phages giving positive signal by the production of XO were selected and further purified. To validate these clones, purified phages were lysogenized in E. coli Y1089 and their lysates were analysed for enzyme activity and immunoreactivity, It was verified that lysates of the purified recombinant phage lysogens exhibited the enzymatic activity as well as bound wi th XO antibody, when induced by IPTG. The above results assert that selected recombinant phage carries mouse liver XO gene.

  • PDF

Browning Degree of Various Apple Cultivars for Minimal Processing (최소 가공용 사과의 품종별 갈변정도)

  • Hong, Jeong Jin;Seol, Hui Gyeong;Jeong, Eun Ho;Kim, Yeong Bong;Hong, Kwang Pyo
    • The Korean Journal of Food And Nutrition
    • /
    • v.31 no.5
    • /
    • pp.689-695
    • /
    • 2018
  • This study was carried out to select apple cultivars suited for the fresh cutting or minimal processing as investigating correlation between total phenol and ascorbic acid as browning factors and the enzymatic browning degree. In soluble solid-acid ratio, 'Gamhong' and 'Arisoo' were highest value of 71.69 and 71.52 compared with other cultivars. And the change of lightness (${\Delta}L$) and color (${\Delta}E$) in 'Gamhong' and 'Arisoo' showed lower than other cultivars while 'Arkansas black' was highest change of lightness and color as ${\Delta}L$ 8.27, ${\Delta}E$ 13.86 respectively. Total phenol and ascorbic acid contents in 'Arkansas black' were highest such as 334.3 mg GAE/100 g and 9.22 mg/100 g, respectively. And polyphenol oxidase (PPO) activity also was the highest such as 14.43unit in 'Arkansas black'. The correlation test showed browning of apple was significantly dependent on ${\Delta}L$, ${\Delta}E$, total phenol, ascorbic acid contents and PPO activity (p<0.01). On the contrary, no significant correlation had been determined between the browning and ascorbic acid oxidase (AAO) activity. Thus, it seemed that the browning of apple was not only due to oxidation of ascorbic acid but also total phenol. It was suggested that 'Gamhong' and 'Arisoo' were suitable for fresh cutting or minimal processing.

Protein-protein Interaction Network Analyses for Elucidating the Roles of LOXL2-delta72 in Esophageal Squamous Cell Carcinoma

  • Wu, Bing-Li;Zou, Hai-Ying;Lv, Guo-Qing;Du, Ze-Peng;Wu, Jian-Yi;Zhang, Pi-Xian;Xu, Li-Yan;Li, En-Min
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.15 no.5
    • /
    • pp.2345-2351
    • /
    • 2014
  • Lysyl oxidase-like 2 (LOXL2), a member of the lysyl oxidase (LOX) family, is a copper-dependent enzyme that catalyzes oxidative deamination of lysine residues on protein substrates. LOXL2 was found to be overexpressed in esophageal squamous cell carcinoma (ESCC) in our previous research. We later identified a LOXL2 splicing variant LOXL2-delta72 and we overexpressed LOXL2-delta72 and its wild type counterpart in ESCC cells following microarray analyses. First, the differentially expressed genes (DEGs) of LOXL2 and LOXL2-delta72 compared to empty plasmid were applied to generate protein-protein interaction (PPI) sub-networks. Comparison of these two sub-networks showed hundreds of different proteins. To reveal the potential specific roles of LOXL2- delta72 compared to its wild type, the DEGs of LOXL2-delta72 vs LOXL2 were also applied to construct a PPI sub-network which was annotated by Gene Ontology. The functional annotation map indicated the third PPI sub-network involved hundreds of GO terms, such as "cell cycle arrest", "G1/S transition of mitotic cell cycle", "interphase", "cell-matrix adhesion" and "cell-substrate adhesion", as well as significant "immunity" related terms, such as "innate immune response", "regulation of defense response" and "Toll signaling pathway". These results provide important clues for experimental identification of the specific biological roles and molecular mechanisms of LOXL2-delta72. This study also provided a work flow to test the different roles of a splicing variant with high-throughput data.

Isolation and Characterization of Nicotine-Degrading Bacterium Arthrobacter sp. NU11 and NU15 (니코틴 분해세균 Arthrobacter sp. NU11과 NU15의 분리 및 특성)

  • Jeong, Yeonju;Oh, Ji-Sung;Roh, Dong-Hyun
    • Korean Journal of Microbiology
    • /
    • v.50 no.1
    • /
    • pp.67-72
    • /
    • 2014
  • Minimal broth containing nicotine as a sole carbon source (MB/N) was used to isolate novel nicotine-degrading bacterial strains from tobacco plants and field soils. Comparative analysis of 16S rRNA gene sequence, phenotypic test and morphological tests showed that the position of these isolates were in the genus Arthrobacter of the family Micrococcaceae. The highest 16S rRNA gene sequence similarity of the isolate NU11 and NU15 to type strains in the genus Arthrobacter were Arthrobacter equi (98.2%) which was presumably a novel strain and Arthrobacter nicotinovorans (99.8%), respectively. Both strain NU11 and NU15 showed rod shaped, Gram-positive characteristics and catalase activity, but did not show oxidase activity. The novel strain NU11 was found to degrade efficiently nicotine in MB/N medium by the analysis of UV absorption spectra and could be used as an organism in bioremediation technique.

Xanthoangelol and 4-Hydroxyderricin Are the Major Active Principles of the Inhibitory Activities against Monoamine Oxidases on Angelica keiskei K

  • Kim, Ji Ho;Son, Yeon Kyung;Kim, Gun Hee;Hwang, Keum Hee
    • Biomolecules & Therapeutics
    • /
    • v.21 no.3
    • /
    • pp.234-240
    • /
    • 2013
  • Monoamine oxidase inhibitors (MAOI) have been widely used as antidepressants. Recently, there has been renewed interest in MAO inhibitors. The activity-guided fractionation of extracts from Angelica keiskei Koidzumi (A. keiskei K.) led to the isolation of two prenylated chalcones, xanthoangelol and 4-hydroxyderricin and a flavonoid, cynaroside. These three isolated compounds are the major active ingredients of A. keiskei K. to inhibit the MAOs and DBH activities. Xanthoangelol is a nonselective MAO inhibitor, and a potent dopamine ${\beta}$-hydroxylase (DBH) inhibitor. $IC_{50}$ values of xanthoangelol to MAO-A and MAO-B were calculated to be 43.4 ${\mu}M$, and 43.9 ${\mu}M$. These values were very similar to iproniazid, which is a nonselective MAO inhibitor used as a drug against depression. The $IC_{50}$ values of iproniazid were 37 ${\mu}M$, and 42.5 ${\mu}M$ in our parallel examination. Moreover, $IC_{50}$ value of xanthoangelol to DBH was calculated 0.52 ${\mu}M$. 4-Hydroxyderricin is a potent selective MAO-B inhibitor and also mildly inhibits DBH activity. The $IC_{50}$ value of 4-hydroxyderricin to MAO-B was calculated to be 3.43 ${\mu}M$ and this value was higher than that of deprenyl (0.046 ${\mu}M$) used as a positive control for selective MAO-B inhibitor in our test. Cynaroside is a most potent DBH inhibitor. The $IC_{50}$ value of cynaroside to DBH was calculated at 0.0410 ${\mu}M$. Results of this study suggest that the two prenylated chalcones, xanthoangelol and 4-hydroxyderricin isolated from A. keiskei K., are expected for potent candidates for development of combined antidepressant drug. A. keiskei K. will be an excellent new bio-functional food material that has the combined antidepressant effect.

Development of Total Cholesterol Detection System by Fluorescence Chromatography (형광 크로마토그래피에 의한 콜레스테롤 측정법의 개발)

  • Oh, Sang-Wook
    • Journal of Food Hygiene and Safety
    • /
    • v.24 no.2
    • /
    • pp.148-153
    • /
    • 2009
  • In this study, we have developed a fluorescence chromatographic assay for the quantification of total cholesterol in serum, which is a well-known risk predictor for cardiovascular diseases. The new assay system consists of a chromatographic strip in a cartridge, enzyme buffer containing cholesterol esterase, cholesterol oxidase, horseradish peroxidase, and color developer AEC, and a laser fluorescence scanner. The correlation coefficient (r) between cholesterol concentration and relative fluorescence units was 0.968 in the new assay, showing a reliable linearity through the tested range of cholesterol. Recovery test and comparability with a Hitachi 747 instrument showed 106.5-94% and r = 0.939 (p<0.001), respectively. The new assay system for cholesterol was developed as a pre-POCT platform conducted in clinics since it is fast (8 min) and uses a small volume of sample ($5\;{\mu}l$), and it may be applied for on-site diagnostics to replace expensive automated biochemical analyzer.