• Title/Summary/Keyword: reactive nitrogen species

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Characterization of an Extracytoplasmic Chaperone Spy in Protecting Salmonella against Reactive Oxygen/Nitrogen Species

  • Park, Yoon Mee;Lee, Hwa Jeong;Bang, Iel Soo
    • International Journal of Oral Biology
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    • v.39 no.4
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    • pp.207-213
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    • 2014
  • Antimicrobial actions of reactive oxygen/nitrogen species (ROS/RNS) derived from products of NADPH oxidase and inducible nitric oxide (NO) synthase in host phagocytes inactivate various bacterial macromolecules. To cope with these cytotoxic radicals, pathogenic bacteria have evolved to conserve systems necessary for detoxifying ROS/RNS and repairing damages caused by their actions. In response to these stresses, bacteria also induce expression of molecular chaperones to aid in ameliorating protein misfolding. In this study, we explored the function of a newly identified chaperone Spy, that is localized exclusively in the periplasm when bacteria exposed to conditions causing spheroplast formation, in the resistance of Salmonella Typhimurium to ROS/RNS. A spy deletion mutant was constructed in S. Typhimurium by a PCR-mediated method of one-step gene inactivation with ${\lambda}$ Red recombinase, and subjected to ROS/RNS stresses. The spy mutant Salmonella showed a modest decrease in growth rate in NO-producing cultures, and no detectable difference of growth rate in $H_2O_2$ containing cultures, compared with that of wild type Salmonella. Quantitative RT-PCR analysis showed that spy mRNA levels were similar regardless of both stresses, but were increased considerably in Salmonella mutants lacking the flavohemoglobin Hmp, which are incapable of NO detoxification, and lacking an alternative sigma factor RpoS, conferring hypersusceptibility to $H_2O_2$. Results demonstrate that Spy expression can be induced under extreme conditions of both stresses, and suggest that the protein may have supportive roles in maintaining proteostasis in the periplasm where various chaperones may act in concert with Spy, thereby protecting bacteria against toxicities of ROS/RNS.

Nonthermal Atmospheric Pressure Plasmas and their Applications to Plasma Bioscience and Medicines

  • Choe, Eun-Ha
    • Proceedings of the Korean Vacuum Society Conference
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    • 2015.08a
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    • pp.56.2-56.2
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    • 2015
  • Nonthermal Atmospheric Pressure Plasmas and their Applications to Plasma Bioscience and Medicines have been introduced for next generation human healthcare's quantum developments. Various kinds of nonthermal atmospheric pressure plasmas have been introduced and their electron temperature and plasma densities along with reactive oxygen and nitrogen species have been diagnosed and analyzed for biological cell interactions, especially, used in Plasma Bioscience Research Center (PBRC), Korea. Herein, we have also introduced the plasma-initiated ultraviolet photolysis, which might be a generation mechanism for the reactive oxygen and nitrogen species (RONS) intracellular and extracellular regions inside the liquid when the plasma has been bombarded onto the water. Finally we have investigated the interactions of these RONS with the various cancer cells resulting in apoptotic cell death.

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Reactive nitrogen metabolism: a novel frontier in plant nitrogen metabolism

  • Sakamoto, Atsushi;Takahashi, Misa;Morikawa, Hiromichi
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.65-70
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    • 2005
  • A growing body of evidence shows that nitric oxide $({\cdot}NO)$ and ${\cdot}NO-derived$ reactive nitrogen species (RNS) act as both plant physiological regulators and stressors. However, very little is known concerning metabolism of RNS in plant cells. In this paper, we explore a plant metabolic basis for RNS, with special emphasis on the possible relationship to nitrogen assimilation, and discuss the potential of the metabolic engineering for plant-biotechnological application.

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Effects of Ichungwhan on the Aging Process (이정환(二精丸)이 노화과정에 미치는 영향)

  • Jeong, Ji-Cheon;Hyun, Min-Kyung
    • The Journal of Internal Korean Medicine
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    • v.26 no.2
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    • pp.379-389
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    • 2005
  • Objectives: It is well known that aging and aging-related diseases are linked to the increased level of oxidative stress caused by reactive oxygen species(ROS) and reactive nitrogen species(RNS). Nonprotein-SH decreases during aging, while substances such as ROS, nitric oxide(NO), peroxynitrite($ONOO^-$), myeloperoxidase(MPO), and dityrosine show a significant increase. This study investigated the effect of Ichungwhan on the aging process by examining its effect on the generation of the above-mentioned substances. Methods: Four comparison groups of SD rats were used. They were 6 month-old rats, 24 month-old rats, and 24 month-old rats fed with food containing 0.1% and 0.3% of Ichungwhan extract. The amount of NO, $ONOO^-$, and ROS in the rats' kidneys were examined using a fluorescence microplate reader. The reagents used for this purpose include: dihydrorhodamine 123 (DHR 123), 2',7' -dichlorodihydrofluorescein, diacetate(DCFDA), and 4,5-diaminofluorescein(DAF-2). A spectrophotometer was used to investigate the reactivity of nonprotein-SH and myeioperoxidase(MPO), using reagents such as trichloroacetic acid(TCA) and tetramethylbenzidine(TMB). The amounts of MPO protein and dityrosine were measued by western blot. Results: The observed effects of Ichungwhan on rats were as follows: increase of nonprotein-SH; effective decrease of RNS level by suppression of the generation system of $ONOO^-$ and NO; decrease of ROS level; decrease of the MPO reactivity and the subsequent reduction of amount of MPO protein; retardation of dityrosine formation. It can be hypothesized, therefore, that Ichungwhan affects both the earlier and later phases of the molecular inflammatory process, and retards the aging process. Conclusions: Empirical evidence in this study supports a role for Ichungwhan in generation mechanisms of aging process-linked substances ROS, NO, $ONOO^-$, nonprotein-SH, MPO and dityrosine. Affects contrary to the aging process observed in rats beg further empiricism to investigate potential application of Ichungwhan as a medication for age-related diseases in humans.

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Preventive Effects of GLEDITSIAE SPINA Ethanol Extracts and its Fraction on Oxidative Stress and Human LDL Oxidation (GLEDITSIAE SPINA 에탄올 추출물 및 분획물이 산화적 스트레스와 human LDL 산화억제에 미치는 영향)

  • Kim, Hyuck;Lee, Min-Ja;Lee, Hye-Sook;Jung, Hyun-Jung;Choi, Sung-Kyu;Lee, Chang-Sub;Park, Won-Hwan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.3
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    • pp.631-638
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    • 2009
  • GLEDITSIAE SPINA (GS) has been used as folk remedies traditionally for treatment of antiphlogistic and antifebrile agents. An ethanol extract and its fraction of GS were assessed to determine the mechanism of its antioxidant activity. Also, inhibitory effect of extract from GS and its fraction measuring the inhibitory effect on $Cu^{2+}$-induced human low-density lipoprotein (LDL) oxidation. GS ethanol extract and its fraction exhibited a concentration-dependent reactive oxygen species (ROS) and reactive nitrogen species (RNS) scavenging activities, including trolox equivalent antioxidant capacity (TEAC), OPPH radical, superoxide anion, hydroxyl radicals, peroxynitrite and nitric oxide, using different assay systems. Furthermore, the GS ethanol extract and its fraction showed dose-dependent protection of LDL oxidation induced by $CuSO_4$. In addition, the GS ethanol extract and its fraction were characterized as containing a high amount of total phenolics. These results suggest that GS ethanol extract and its fraction might be helpful for preventing oxidative stress and protecting LDL oxidation.

Enhancement of Nitric Oxide with nonthermal plasma jet and its effect on Escherichia coli inactivation

  • Shaw, Priyanka;Kumar, Naresh;Attri, Pankaj;Kwak, Hyong Sin;Choi, Eun Ha
    • Proceedings of the Korean Vacuum Society Conference
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    • 2015.08a
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    • pp.159-159
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    • 2015
  • A new approach for antimicrobial is based on the overproduction of reactive nitrogen species (RNS), especially; nitric oxide (NO) and peroxinitrite (ONOO-) are important factors to deactivate the bacteria. Recently, non-thermal atmospheric pressure plasma jet (APPJ) has been frequently used in the field of microbial sterilization through the generation of different kinds of RNS/ROS species. However, in previous study we showed APPJ has combine effects ROS/RNS on bacterial sterilization. It is not still clear whether this bacterial killing effect has been done through ROS or RNS. We need to further investigate separate effect of ROS and RNS on bacterial sterilization. Hence, in this work, we have enhanced NO production, especially; by applying a 1% of HNO3 vapour to the N2 based APPJ. In comparison with nitrogen plasma with inclusion of water vapour plasma, it has been shown that nitrogen plasma with inclusion of 1% of HNO3 vapour has higher efficiency in killing the E. coli through the high production of NO. We also investigate the enhancement of NO species both in atmosphere by emission spectrum and inside the solution by ultraviolet absorption spectroscopy. Moreover, qPCR analysis of oxidative stress mRNA shows higher gene expression. It is noted that 1% of HNO3 vapour plasma generates high amount of NO for killing bacteria.

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Model Development for the Nitrification-Denitrification Coupled Process

  • Lee, Mee-Sun;Lee, Kang-Kun
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2002.04a
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    • pp.195-198
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    • 2002
  • Nitrogen pollution in urban and rural groundwater is a common problem and poses a major threat to drinking water supplies based on groundwater. In this work, the kinetics of nitrification-denitrification coupled reactions are modeled and new reaction modules for the RT3D code (Clement, 1997) describing the fate and transport nitrogen species, dissolved oxygen, dissolved organic carbon, and biomass are developed. The proposed nitrogen transformations and transport model showed very good match with results of a conceptual model. However, the model simulation results for the major reactive species should be tested for validation using experimental and field data.

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Feed Gas Dependent Nonthermal Plasma Interaction with Bio-organisms

  • Baik, Ku-Youn;Park, Gyung-Soon;Kim, Yong-Hee;Yoo, Young-Hyo;Lee, Jin-Young;Choi, Eun-Ha
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.02a
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    • pp.174-174
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    • 2012
  • The nature of feed gas is essential for the active species formed in the nonthermal plasma jets, which would induce various biological phenomena. We investigated the different physiological effects of atmospheric pressure soft-plasma jets on Esherichia coli and blood cells according to the feed gas. Cell death rate, growth curve, membrane molecular changes and induced genes were examined. The relationship between cellular reactions and active species generated by discharge will be discussed.

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Characteristics of Atmosphere-rice Paddy Exchange of Gaseous and Particulate Reactive Nitrogen in Terms of Nitrogen Input to a Single-cropping Rice Paddy Area in Central Japan

  • Hayashi, Kentaro;Ono, Keisuke;Matsuda, Kazuhide;Tokida, Takeshi;Hasegawa, Toshihiro
    • Asian Journal of Atmospheric Environment
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    • v.11 no.3
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    • pp.202-216
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    • 2017
  • Nitrogen (N) is an essential macronutrient. Thus, evaluating its flows and stocks in rice paddy ecosystems provides important insights into the sustainability and environmental loads of rice production. Among the N sources of paddy fields, atmospheric deposition and irrigation inputs remain poorly understood. In particular, insufficient information is available for atmosphere-rice paddy exchange of gaseous and particulate reactive N (Nr, all N species other than molecular N) which represents the net input or output through dry deposition and emission. In this study, we assessed the N inputs via atmospheric deposition and irrigation to a Japanese rice paddy area by weekly monitoring for 2 years with special emphasis on gas and particle exchange. The rice paddy during the cropping season acted as a net emitter of ammonia ($NH_3$) to the atmosphere regardless of the N fertilizer applications, which reduced the effects of dry deposition to the N input. Dry N deposition was quantitatively similar to wet N deposition, when subtracting the rice paddy $NH_3$ emissions from N exchange. The annual N inputs to the rice paddy were 3.2 to $3.6\;kg\;N\;ha^{-1}\;yr^{-1}$ for exchange, 8.1 to $9.8\;kg\;N\;ha^{-1}\;yr^{-1}$ for wet deposition, and 11.1 to $14.5\;kg\;N\;ha^{-1}\;yr^{-1}$ for irrigation. The total N input, 22.8 to $27.5\;kg\;N\;ha^{-1}\;yr^{-1}$, corresponded to 38% to 55% of the N fertilizer application rate and 53% to 67% of the brown rice N uptake. Monitoring of atmospheric deposition and irrigation as N sources for rice paddies will therefore be necessary for adequate N management.

Effect of Antioxidant Supplementation in Freezing Extender on Porcine Sperm Viability, Motility and Reactive Oxygen Species

  • Park, Sang-Hyoun;Yu, Il-Jeoung
    • Journal of Embryo Transfer
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    • v.32 no.1
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    • pp.9-15
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    • 2017
  • The present study was aimed to determine the effect of green tea extract (GTE) and beta-mercaptoethanol (${\beta}$-ME) supplementation in boar sperm freezing extender on sperm motility, viability and reactive oxygen species (ROS) level. Experimental groups were allocated into Lactose-egg yolk (LEY) without antioxidant (control), GTE (1,000 mg/L GTE in LEY) and ${\beta}$-ME ($50{\mu}M$ ${\beta}$-ME in LEY). Spermatozoa extended with LEY were cooled to $5^{\circ}C$ for 3 h and then kept at $5^{\circ}C$ for 30 min following dilution with LEY containing 9% glycerol and 1.5% Equex STM (final sperm concentration: $1{\times}10^8/mL$). Spermatozoa were loaded into straws and frozen in nitrogen vapor for 20 min. Following thawing at $37^{\circ}C$ for 25 sec, sperm viability and ROS level were measured using fluorescent double stain Fertility(R) and cytometry, respectively. Motility and viability of GTE supplemented-group were higher than those of control and ${\beta}$-ME without significance. ROS level in GTE group showed significantly lower than control (P < 0.05). In conclusion, GTE supplementation in boar sperm freezing extender can reduce ROS generation during freezing.