• Title/Summary/Keyword: oxidase test

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Development and Validation of Real-time PCR to Determine Branchiostegus japonicus and B. albus Species Based on Mitochondrial DNA (Real-time PCR 분석법을 이용한 옥돔과 옥두어의 종 판별법 개발)

  • Chung, In Young;Seo, Yong Bae;Yang, Ji-Young;Kim, Gun-Do
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1331-1339
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    • 2017
  • DNA barcoding is the identification of a species based on the DNA sequence of a fragment of the cytochrome C oxidase subunit I (COI) gene in the mitochondrial genome. It is widely applied to assist with the sustainable development of fishery-product resources and the protection of fish biodiversity. This study attempted to verify horse-head fish (Branchiostegus japonicus) and fake horse-head fish (Branchiostegus albus) species, which are commonly consumed in Korea. For the validation of the two species, a real-time PCR method was developed based on the species' mitochondrial DNA genome. Inter-species variations in mitochondrial DNA were observed in a bioinformatics analysis of the mitochondrial genomic DNA sequences of the two species. Some highly conserved regions and a few other regions were identified in the mitochondrial COI of the species. In order to test whether variations in the sequences were definitive, primers that targeted the varied regions of COI were designed and applied to amplify the DNA using the real-time PCR system. Threshold-cycle (Ct) range results confirmed that the Ct ranges of the real-time PCR were identical to the expected species of origin. Efficiency, specificity and cross-reactivity assays showed statistically significant differences between the average Ct of B. japonicus DNA ($21.85{\pm}3.599$) and the average Ct of B. albus DNA ($33.49{\pm}1.183$) for confirming B. japonicus. The assays also showed statistically significant differences between the average Ct of B. albus DNA ($22.49{\pm}0.908$) and the average Ct of B. japonicus DNA ($33.93{\pm}0.479$) for confirming B. albus. The methodology was validated by using ten commercial samples. The genomic DNA-based molecular technique that used the real-time PCR was a reliable method for the taxonomic classification of animal tissues.

Effect of Gamma Irradiation for Hygienic Long-Term Storage on Biological Activity of Teucrium veronicoides (위생적인 장기 보존을 위한 감마선 조사가 곽향(Teucrium veronicoides)의 생리활성에 미치는 영향)

  • Park, Hye-Jin;Park, Ki-Tae;Cho, Young-Je
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.5
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    • pp.581-591
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    • 2017
  • The purpose of this study was to examine the biological activities of gamma-irradiated Teucrium veronicoides. In photostimulated luminescence analysis, non-irradiated sample showed lower than 700 photon counts (PCs), whereas irradiated (5 and 10 kGy) samples showed higher than 700 PCs. The thermoluminescence ratio of non-irradiated samples was less than 0.1, whereas the values of irradiated samples were greater than 0.1. Electron spin resonance analysis was performed confirmed for irradiation identification. The total phenolic contents of hot-water and 50% ethanol extracts were higher than those values after irradiation at 10 kGy. Regarding 1,1-diphenyl-2-picrylhydrazyl radical inhibitory activity, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity, antioxidant protection factor, thiobarbituric acid reactive substance inhibitory activity as antioxidant test and xanthine oxidase inhibitory activity, the effect of gamma irradiation had on significant effects. On the other hand, ${\alpha}-amylase$ inhibitory activity of 10 kGy-irradiated hot-water extract was 23.6% higher than that of the non-irradiated sample. Thus, gamma irradiation could be used for the long-term storage of Teucrium veronicoides.

A Study on the Cosmeceutical activities of Prunus Sargentii R. (산벚나무(Prunus sargentii R.) 수피의 화장품활성에 관한 연구)

  • Park, Jung-Mi;Lee, Jin-Young;Park, Tae-Soon;Hyun, Sok-Jun;Kim, Han-Hyuk;Cho, Young-Je;Kwon, O-Jun;Son, Ae-Ryang;Kim, Dong-Seok;An, Bong-Jeun
    • Applied Biological Chemistry
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    • v.51 no.1
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    • pp.70-78
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    • 2008
  • Prunus sargentii R. of Rosaceae familiy, has been reported to have radical scavenging activity and anti-inflammatory effect. On these facts, biological activity and safety test were conducted to evaluate biological activities of the extracts of P. sargentii R. as a potential pharmaceutical ingredient. The electron donating ability of its ethanol extracts at a 500 ppm level showed 92%, which was higher than that of hot water extract (59%), the superoxide dismutase (SOD)-like activity of the water extract of P. sargentii R. was about 50%, the ethanol extract of P. sargentii R. was about 40% at 1,000 ppm concentration. Xanthine oxidase inhibition by the water extract of P. sargentii R. was about 40% and that by the ethanol extract was 60% respectively at 500 ppm concentration. From the measurement on lipid oxidation, the $Cu^{2+}$ chelating effect of the ethanol extract was higher than that of hot water extract. The $Fe^{2+}$ chelating effect was also shown to be about 80% at a 500 ppm concentration in both hot water extract and ethanol extract. The tyrosinase inhibition effect related to skin-whitening was 26% by hot water extract and 20% by ethanol extract respectively at a 1,000 ppm. Hyaluronidase inhibition activity related to the anti-inflammation effect was 96% in ethanolic extract at a 500 ppm. Clear zones formed by P. sargentii R. against the human skin-resident micro-flora such as Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli and Propionibacterium acnes indicated that antimicrobial activity of the ethanol extract was higher than that of the hot water extract.

Isolation and Identification of a Histamine-degrading Barteria from Salted Mackerel (자반고등어에서 histamine 분해능을 가진 세균의 분리 동정)

  • Hwang Su-Jung;Kim Young-Man
    • Journal of Life Science
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    • v.15 no.5 s.72
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    • pp.743-748
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    • 2005
  • Histamine can be produced at early spoilage stage through decarboxylation of histidine in red-flesh fish by Proteus morganii, Hafnia alvei or Klebsiella pneumoniae. Allergic food poisoning is resulted from the histamine produced when the freshness of Mackerel degrades. Conversely it has been reported that there are bacteria which decompose histamine at the later stage. We isolated histamine decomposers from salted mackerel and studied the characteristics to help establish hygienic measure to prevent outbreak of salted mackerel food poisoning. All the samples were purchased through local supermarket. Histamine decomposers were isolated using restriction medium using histamine 10 species were selected. Identification of these isolates were carried out by the comparison of 16S rDNA partial sequence; as a result, we identified Pseudomonas putida strain RA2 and Halomonas marina, Uncultured Arctic sea ice bacterium clone ARKXV1/2-136, Halomonas venusta, Psychrobacter sp. HS5323, Pseudomonas putida KT2440, Rhodococcus erythropolis, Klebsiella terrigena (Raoultella terrigena), Alteromonadaceae bacterium T1, Shewanella massilia with homology of $100\%,{\;}100\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}99\%,{\;}100\%,{\;}95\%,{\;}99\%,{\;}and{\;}100\%$respectively. Turbidometry determination method and enzymic method were employed to determine the ability of histamine decomposition. Among those species Shewanella massilia showed the highest in ability of histamine decomposition. From these results we confirmed various histamine decomposer were present in salted mackerel product in the market.

Antioxidant Activities of the Mycelium and Culture Broth of Phellinus igniarius and Agrocybe cylindracea (말똥진흙버섯 및 버들송이버섯의 균사체 및 배양액의 항산화활성)

  • Lee, Hang-Woo;Lee, Dong-Woo;Ha, Hyo-Cheol;Jung, In-Change;Lee, Jae-Sung
    • The Korean Journal of Mycology
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    • v.30 no.1
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    • pp.37-43
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    • 2002
  • The fractions obtained from the mycelia and their culture broth of Agrocybe cylindracea and Phellinus igniarius were tested for the antioxidation activities. In electron donating ability test, strong activities more than 70% were observed in ethyl acetate and butanol fractions of A. cylindracea mycelium, butanol fraction of A. cylindracea culture broth, diethyl ether fraction of P. igniarius mycelium, and diethyl ether and ethyl acetate fractions of P. igniarius culture broth. Lipid peroxidation inhibition was observed in diethyl ether, butanol and water fractions of A. cylindracea mycelium, and water fraction of A. cylindracea induced the decrease in uric acid production. Activities of $50{\sim}60\;%$ of nitrite scavenging ability were observed in diethyl ether fraction of A. cylindracea mycelium, ethyl acetate and water fractions of A. cylindracea culture broth, diethyl ether fraction of P. igniarius mycelium, and diethyl ether, chloroform, butanol and water fractions of P. igniarius culture broth. In melanin biosynthesis inhibition activity measurement, water fraction of A. cylindracea mycelium and butanol fraction of P. igniarius mycelium showed twice higher activities than control. From these results, the fungi of P. igniarius and A. cylindracea had strong antioxidant activities.

Desmutagenicity of the Enzymatic Browning Reaction Products Which Obtained from Prunus salicina (yellow) Enzyme and Polyphenol Compounds (재래종 황색자두효소 갈변반응 생성물의 돌연변이 억제작용)

  • Ham, Seung-Shi
    • Applied Biological Chemistry
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    • v.30 no.1
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    • pp.71-76
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    • 1987
  • The mutagenicity and desmutagenicity on enzymatic browning reaction products which obtained from prunes salicina (yellow) enzyme and polyphenol compounds were carried out. In the rec-assay on Bacillus subtilis strains H17 and M45, the enzymatic browning reaction products of pyrogallol, hydroxyhydroquinone, 3,4-dihydroxytoluene and catechol of $10^{-2}M$ did not showed mutagenicity. In the effects of various metal ions on the rec-assay, the enzymatic browning reaction products of pyrogallol showed mutagenic activity by $Fe^{3+},\;Mn^{2+},\;Zn^{2+},\;Ni^{2+}$ and $Al^{3+}$. In the enzymatic browning reaction products of hydroxyhydroquinone, $Cu^{2+},\;Mn^{2+}$ and $Pb^{2+}$ were effected in mutagenic action and the enzymatic browning reaction products of catechol was effected in mutagenic action by $Mn^{2+}$. In the DNA-breaking action of enzymatic browning reaction products of pyrogallol, hydroxyhydroquinone, 3,4-dihyroxytoluene and catechol did not show, DNA-breaking action. In the effects of various metal ions on the DNA-breaking action of enzymatic browning reaction products, $Cu^{2+}$ showed DNA-breaking action. In the mutagenicity test on Sal. typhimurium strains TA98 and TA 100 with S-9 mix, 4 kinds of browned substances did sot shove muragenicity, all the browned substances showed strong desmutagenic activity in the presence of benzo $({\alpha})-pyrene$ with S-9 mix.

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Anti-tumor Effects of Penfluridol through Dysregulation of Cholesterol Homeostasis

  • Wu, Lu;Liu, Yan-Yang;Li, Zhi-Xi;Zhao, Qian;Wang, Xia;Yu, Yang;Wang, Yu-Yi;Wang, Yi-Qin;Luo, Feng
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.489-494
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    • 2014
  • Background: Psychiatric patients appear to be at lower risk of cancer. Some antipsychotic drugs might have inhibitory effects on tumor growth, including penfluridol, a strong agent. To test this, we conducted a study to determine whether penfluridol exerts cytotoxic effects on tumor cells and, if so, to explore its anti-tumor mechanisms. Methods: Growth inhibition of mouse cancer cell lines by penfluridol was determined using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Cytotoxic activity was determined by clonogenic cell survival and trypan blue assays. Animal tumor models of these cancer cells were established and to evaluate penfluridol for its anti-tumor efficacy in vivo. Unesterified cholesterol in cancer cells was examined by filipin staining. Serum total cholesterol and tumor total cholesterol were detected using the cholesterol oxidase/p-aminophenazone (CHOD-PAP) method. Results: Penfluridol inhibited the proliferation of B16 melanoma (B16/F10), LL/2 lung carcinoma (LL/2), CT26 colon carcinoma (CT26) and 4T1 breast cancer (4T1) cells in vitro. In vivo penfluridol was particularly effective at inhibiting LL/2 lung tumor growth, and obviously prolonged the survival time of mice bearing LL/2 lung tumors implanted subcutaneously. Accumulated unesterified cholesterol was found in all of the cancer cells treated with penfluridol, and this effect was most evident in LL/2, 4T1 and CT26 cells. No significant difference in serum cholesterol levels was found between the normal saline-treated mice and the penfluridol-treated mice. However, a dose-dependent decrease of total cholesterol in tumor tissues was observed in penfluridol-treated mice, which was most evident in B16/F10-, LL/2-, and 4T1-tumor-bearing mice. Conclusion: Our results suggested that penfluridol is not only cytotoxic to cancer cells in vitro but can also inhibit tumor growth in vivo. Dysregulation of cholesterol homeostasis by penfluridol may be involved in its anti-tumor mechanisms.

Skin Anti-aging Effect of Forsythia viridissima L. Extract (연교추출물의 피부 항노화 효과)

  • Kim, Mi-Jin;Kim, Ja-Young;Jung, Teak-Kyu;Choi, Sang-Won;Yoon, Kyung-Sup
    • KSBB Journal
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    • v.21 no.6 s.101
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    • pp.444-450
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    • 2006
  • Skin anti-aging effect of Forsythia viridissima L. extract was evaluated by using antioxidant assay, expression of type I procollagen, and UVA-induced matrix metalloproteinase-1 in human dermal fibroblasts. Matairesinol-rich Forsythia viridissima L. extract was showed the scavenging activity of radicals and reactive oxygen species with the $IC_{50}$ values of $4.50\;{\mu}m/ml$ against 1,1-diphenyl-2-picrylhydrazly radical and $542.43\;{mu}m/ml$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. The type I procollagen was increased 33.76% by treatment with matairesinol-rich Forsythia viridissima L. extract, and UVA-induced MMP-1 was reduced 35.78% in a dose dependent manner. In the human skin irritation test, 2% matairesinol-rich Forsythia viridissima L. extract did not show any adverse effect. Also, the clinical study indicated that a cream group treated with 0.2% matairesinol-rich Forsythia viridissima L. extract significantly reduced skin wrinkles, as compared with a non-treated cream group (p < 0.05). These results suggest that Forsythia viridissima L. extract may be useful as a potential source of functional anti-aging cosmetics.

Biological Evaluation of the Methanolic Extract of Eriobotrya japonica and Its Irradiation Effect (비파 메탄올 추출물의 생리활성 및 방사선 조사 효과)

  • Kim, Hee-Jung;Jo, Cheor-Un;Kim, Tae-Hoon;Kim, Dong-Sup;Park, Moon-Young;Byun, Myung-Woo
    • Korean Journal of Food Science and Technology
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    • v.38 no.5
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    • pp.684-690
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    • 2006
  • Eriobotrya japonica has been used as a folk medicine for treatment of skin diseases, inflammation, coughing, phlegm, and ulcers in Korea and other Asian countries. In a search for possible bioactive agents from natural sources, we found that the methanolic extracts from various parts of E. japonica showed moderate antioxidative and antimicrobial activities in several in vitro bioassay systems. Additionally, the respective parts of E. japonica were irradiated at 20 kGy to investigate the effects of irradiation. Gamma irradiation of E. japonica extracts removed the deep greenish color without affecting its natural biological activities such as its antioxidative and antimicrobial properties. Based on these findings, the methanolic extracts of this plant source may be not affected by gamma irradiation as its bioactive constituents may be insensitive to this irradiation. Moreover, the methanolic extract of E. japonica may serve as a good natural resource for beneficial functions in food and other related industries.

Effects of Reactive Oxygen Species on Sperm Function, Lipid Peroxidation and DNA Fragmentation in Bovine Spermatozoa (소 정자에 있어서 활성산소계가 정자 기능과 지방산화 및 DNA 절편화에 미치는 영향)

  • Ryu, Buom-Yong;Chung, Yung-Chai;Kim, Chang-Keun;Shin, Hyun-A;Han, Jung-Ho;Pang, Myung-Geol;Oh, Sun-Kyung;Kim, Seok-Hyun;Moon, Shin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.2
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    • pp.105-115
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    • 2002
  • Objective : To evaluate the effects of the reactive oxygen species (ROS) generated with a xanthine (X) and xanthine oxidase (XO) system on sperm function, the change of sperm characteristics, lipid peroxidation, and DNA fragmentation in bovine spermatozoa. Materials and Methods: ROS were produced using a combination of 1000 uM X and 50 mU/ml XO. The ROS scavengers: superoxide dismu tase (SOD) (200 U/ml) and catalase (500 U/ml) were also tested. Spermatozoa were incubated for 2 hours in BWW medium with a combination of X-XO supplemented with or without ROS scavengers at $37^{circ}C$ under 5% $CO_2$ incubator. Sperm movement characteristics by CASA (computer-aided sperm analysis), HOST (hypoosmotic swelling test), Caionophore induced acrosome reaction, malondialdehyde formation for the analysis of lipid peroxidation, the percentage of DNA fragmentation using the method of TdT-mediated nick end labelling (TUNEL) by flow cytometry were determined after 2 hours incubation. Results: The action of ROS on bovine spermatozoa resulted in a decreased in capacity for sperm motility, Ca-ionophore induced acrosome reaction and membrane integrity, an increased in malondialdehyde formation and the percentage of sperm with DNA fragmentation. In the effects of antioxidant, catalase completely alleviated the toxic effects induced by the ROS in terms of sperm function and characteristics, however SOD exhibited no capacity to reduce the toxic effects. Conclusion: The ROS can induce significant damages to sperm functions and characteristics. The useful ROS scavengers can minimized the defects of sperm function and various damages of spermatozoa.