Detection of Inducible Nitric Oxide Synthase Using a Suite of Electrochemical, fluorescence, and Surface Plasmon Resonance Biosensors ⇑ Naumih M

Total Page:16

File Type:pdf, Size:1020Kb

Detection of Inducible Nitric Oxide Synthase Using a Suite of Electrochemical, fluorescence, and Surface Plasmon Resonance Biosensors ⇑ Naumih M Analytical Biochemistry 413 (2011) 157–163 Contents lists available at ScienceDirect Analytical Biochemistry journal homepage: www.elsevier.com/locate/yabio Detection of inducible nitric oxide synthase using a suite of electrochemical, fluorescence, and surface plasmon resonance biosensors ⇑ Naumih M. Noah, Saamia Alam, Omowunmi A. Sadik Center for Advanced Sensors and Environmental Systems (CASE), Department of Chemistry, State University of New York at Binghamton, Binghamton, NY 13902, USA article info abstract Article history: A suite of biosensors for rapid detection of inducible nitric oxide synthase (iNOS) is described. First, a Received 17 December 2010 metal-enhanced electrochemical detection (MED) sensor, which relied on the redox properties of a silver Received in revised form 4 February 2011 monolayer, was developed. The linear detection range was between 8.64 Â 10À2 and 5.4 Â 101 ng/ml Accepted 7 February 2011 with a detection limit of 1.69 Â 10À4 ng/ml. This method was compared with surface plasmon resonance Available online 1 April 2011 (SPR) biosensors in which polyclonal mouse anti-iNOS was covalently immobilized onto a gold surface using an iNOS antigen. The linear detection range recorded was between 3.37 Â 101 and 5.4 Â 10À2 ng/ Keywords: ml with a detection limit of 2 Â 10À3 ng/ml. Finally, an ultrasensitive portable capillary (UPAC) fluores- Inducible nitric oxide synthase cence immunosensor, in which a mouse anti-iNOS antibody was covalently immobilized onto the inner Metal-enhanced detection Surface plasmon resonance surface of a capillary and a rabbit anti-iNOS antibody was employed as the secondary antibody, was Ultrasensitive portable capillary sensor developed. The resulting signals were found to be directly proportional to iNOS concentrations between 1 2 3 Pain biomarkers 1.52 Â 10À and 1.52 Â 10À ng/ml with a detection limit of 1.05 Â 10À ng/ml. These immunosensors exhibit low cross-reactivity toward potential interferents such as human serum albumin and ovalbumin. The SPR and UPAC biosensors were validated using simulated blood spiked with recombinant iNOS, resulting in recoveries of 85% and 88.5%, respectively. The research presented in this article could poten- tially provide new ways of detecting NO for diagnostic and biomarker purposes in medical research. Ó 2011 Elsevier Inc. All rights reserved. Nitric oxide (NO)1 is a highly reactive free radical and an impor- other hand, overproduction of NO often leads to its role as a major tant intra- and intercellular signaling molecule used for the regula- cytotoxic mediator in pathological processes, especially in inflamma- tion of diverse physiological and pathophysiological mechanisms tory disorders. Excess NO (>10 lM) interacts with oxygen radicals to such as cardiovascular, nervous, and immunological systems [1,2]. form highly reactive peroxynitrite, which in turn induces inflamma- Also, NO has contrasting roles in living organisms; it acts as a biolog- tory cellular cytokines and hence cytokine-induced cell death via ical mediator similar to neurotransmitters regulating the blood ves- apoptosis and necrosis [2,3]. sels as well as an important host defense effector in the immune NO is synthesized in vivo during the conversion of L-arginine to system. At small concentrations (62 lM), it acts as an intra- or inter- citrulline by nitric oxide synthase (NOS). It occurs in different iso- cellular second messenger, usually via guanylate cyclase [2]. On the forms with different properties depending on the role of NO to be synthesized. For example, endothelial NOS (eNOS) and neuronal NOS (nNOS) are constitutive enzymes and are constantly ex- ⇑ pressed, even at rest in the blood vessels and the brain neurons Corresponding author. Fax: +1 607 777 447. [4]. In contrast, inducible NOS (iNOS) is expressed in macrophages E-mail address: [email protected] (O.A. Sadik). and other tissues in response to infection or inflammation [5].Itis 1 Abbreviations used: NO, nitric oxide; NOS, nitric oxide synthase; iNOS, inducible NOS; UPAC, ultrasensitive portable capillary; MED, metal-enhanced electrochemical expressed after an exposure to diverse stimuli such as the detection; BSA, bovine serum albumin; NaH2PO4, sodium dihydrogen phosphate; inflammatory cytokines and lipopolysaccharides (LPSs) [6]. Once Na2HPO4, disodium hydrogen phosphate; Na2CO3, sodium carbonate; NaHCO3, expressed, iNOS generates significantly large and sustained sodium hydrogen carbonate; IgG, immunoglobulin G; Tris–HCl, Tris–hydrochloric amounts of NO in the blood, resulting in pathological effects acid; HSA, human serum albumin; DDAO, 9H-(1,3-dichloro-9,9-dimethylacridin- 2-one-7-yl); PNPP, paranitrophenyl phosphate; GMBS, c-maleimido butyryloxy suc- [6,7]. The production of iNOS is a nonspecific event because it cinimide ester; NaCl, sodium chloride; H2SO4, sulfuric acid; NaOH, sodium hydroxide; can occur in a variety of cell types. For example, an increased pro- DMSO, dimethyl sulfoxide; HCl, hydrochloric acid; EDC, 1-ethyl-3-(3-dimethyl duction of NO has been observed during inflammation and arthri- aminopropyl) carbodiimide; NHS, N-hydroxysuccinimide; MgCl2, magnesium chlo- tis; hence, iNOS can be considered as a pain biomarker. It has also ride; PBS, phosphate-buffered saline; EDTA, ethylenediaminetetraacetic acid; DEA, been found to overexpress cyclooxygenase-2 (COX-2), which is an- diethanolamine; ELISA, enzyme-linked immunosorbent assay; PBS-T, PBS containing 0.05% Tween 20; OD, optical density; Ab–Ag, antibody–antigen; SPR, surface plasmon other pain biomarker. These two major pain biomarkers have been resonance; UPD, underpotential deposition; SAM, self-assembled monolayer. found to be expressed in cancer cells [8–10]. 0003-2697/$ - see front matter Ó 2011 Elsevier Inc. All rights reserved. doi:10.1016/j.ab.2011.02.010 158 Detection of inducible nitric oxide synthase / N.M. Noah et al. / Anal. Biochem. 413 (2011) 157–163 There are numerous approaches in the literature for studying and 1.0 mM ethylenediaminetetraacetic acid (EDTA), and PBSB NO. These include chemiluminescence, paramagnetic resonance consisted of PBS and 1% BSA. All PBS buffer solutions were adjusted spectrometry, resonance imaging, and bioassays. We previously re- to pH 7.6 using concentrated phosphoric acid or concentrated ported on the development of an inexpensive amperometric sensor NaOH. Tris–HCl buffer solution was prepared using 6.3509 g of for the determination of the inhibitory effects of NO on xanthine Tris–HCl and 7.2199 g of Tris base and the pH adjusted to 8.0 using oxidase, horseradish peroxidase, polyphenol oxidase, and glucose HCl or 10 M NaOH. Carbonate buffer (pH 9.6) was prepared using oxidase [11–13]. Recently, Njagi et al. [14] reported an electro- 0.1 M Na2CO3 and 0.1 M NaHCO3 and the pH adjusted to 9.6 by chemical sensor for monitoring NO in brain slices. Several micro- titrating the Na2CO3 with NaHCO3 via mixing. PNPP solution was electrodes based on either direct or catalytic oxidation of NO prepared by dissolving two PNPP tablets in 0.5 M diethanolamine have also been developed [15]. Other NO sensors developed have (DEA) buffer (pH 9.8) containing MgCl2. used metalloporphyrins and platinized platinum to provide an electrocatalytic oxidation of NO [16–18]. However, these methods Methods have been found to have poor sensitivity and selectivity and also lack stability in simulated biological samples [14,15,17]. In addi- ELISA tion, NO is highly reactive with a very short half-life (5 s), thereby The enzyme-linked immunosorbent assay (ELISA) protocol used making it difficult to be continuously monitored in real time and was based on a method described elsewhere [19]. Briefly, flat- in vivo. bottomed polystyrene 96-well microplates were coated overnight In this work, we propose to monitor iNOS as a viable surrogate at 4 °C with 100 ll/well of 1 lg/ml mouse anti-iNOS polyclonal for the short-lived species, NO. The goal of this study was to devel- antibody (prepared in carbonate buffer, pH 9.6) as the capture op and assess three different biosensors for rapid detection and antibody. The plate was washed three times with PBS containing characterization of iNOS. A suite of advanced biosensors that will 0.05% Tween 20 (PBS-T, pH 7.6), and the same washing procedure enable rapid, portable, autonomous, and real-time sampling of was followed at each subsequent stage of the assay. The plates iNOS detection capabilities is envisioned. Toward this end, we ini- were incubated overnight at 4 °C with 200 ll of 1 mg/ml BSA pre- tiated the development of portable immunosensors, ultrasensitive pared in PBS (pH 7.6). The plates were washed before incubating portable capillary (UPAC) and metal-enhanced electrochemical them at 4 °C with 100 ll of different concentrations of iNOS in detection (MED), for on-demand diagnostics. Comparative analysis PBS–BSA buffer overnight at 4 °C. After washing, 100 llof1lg/ of these biosensors is reported here. ml rabbit anti-iNOS polyclonal antibody was applied to the wells (except the blank). Following incubation for 2 h at ambient tem- peratures, the plates were washed and 100 ll of goat anti-rabbit Materials and methods IgG–alkaline phosphatase in Tris buffer (pH 8.0) was added (excluding the blanks), followed by an additional 1 h of incubation Materials at room temperature. After final rinsing with Tris buffer (pH 8.0), 100 ll of 1 mg/ml PNPP solution prepared in 10% DEA buffer was iNOS and rabbit anti-iNOS polyclonal antibody were purchased finally added and the plates were incubated at room temperature from Cayman Chemical (Ann Arbor, MI, USA) and stored at À20 °C. for 30 min. Optical densities (ODs) of the solutions were then mea- Mouse anti-iNOS polyclonal antibody was purchased from Abcam sured at 405 nm using a Synergy HTRDR multidetection microplate (Cambridge, MA, USA). Bovine serum albumin (BSA) was reader. purchased from Thermo Fisher Scientific (Pittsburgh, PA, USA). The following reagents were purchased from Sigma–Aldrich (St.
Recommended publications
  • Characterization of Polyphenol Oxidase and Antioxidants from Pawpaw (Asimina Tribola) Fruit
    University of Kentucky UKnowledge University of Kentucky Master's Theses Graduate School 2007 CHARACTERIZATION OF POLYPHENOL OXIDASE AND ANTIOXIDANTS FROM PAWPAW (ASIMINA TRIBOLA) FRUIT Caodi Fang University of Kentucky, [email protected] Right click to open a feedback form in a new tab to let us know how this document benefits ou.y Recommended Citation Fang, Caodi, "CHARACTERIZATION OF POLYPHENOL OXIDASE AND ANTIOXIDANTS FROM PAWPAW (ASIMINA TRIBOLA) FRUIT" (2007). University of Kentucky Master's Theses. 477. https://uknowledge.uky.edu/gradschool_theses/477 This Thesis is brought to you for free and open access by the Graduate School at UKnowledge. It has been accepted for inclusion in University of Kentucky Master's Theses by an authorized administrator of UKnowledge. For more information, please contact [email protected]. ABSTRACT OF THESIS CHARACTERIZATION OF POLYPHENOL OXIDASE AND ANTIOXIDANTS FROM PAWPAW (ASIMINA TRIBOLA) FRUIT Crude polyphenol oxidase (PPO) was extracted from pawpaw (Asimina triloba) fruit. The enzyme exhibited a maximum activity at pH 6.5–7.0 and 5–20 °C, and had -1 a maximum catalysis rate (Vmax) of 0.1363 s and a reaction constant (Km) of 0.3266 M. It was almost completely inactivated when incubated at 80 °C for 10 min. Two isoforms of PPO (MW 28.2 and 38.3 kDa) were identified by Sephadex gel filtration chromatography and polyacrylamide gel electrophoresis. Both the concentration and the total activity of the two isoforms differed (P < 0.05) between seven genotypes of pawpaw tested. Thermal stability (92 °C, 1–5 min) and colorimetry (L* a* b*) analyses showed significant variations between genotypes.
    [Show full text]
  • 3. Methodology
    3. METHODOLOGY The present study focused on the responses evoked by B. monnieri under conditions of oxidative stress. The plan of work and the methodology adopted are presented in this chapter. The work was carried out in four phases. In Phase I, different parts of B. monnieri , namely leaves, stolon and roots, were assessed for their antioxidant potential and free radical scavenging activity. The biomolecular protective effects and the apoptosis- modulating effects of the leaf extract in cell free and in vitro systems were studied in Phase II. Phase III involved assessing the antioxidant status in precision-cut liver slices exposed to oxidative stress and confirming the results in experimental animals. In the final phase of the study, phytochemical analysis of the active component in B. monnieri leaves was carried out. The layout of the study is presented below. PHASE I In this phase, the antioxidant status in different parts of B. monnieri was assessed. The leaves, stolon and roots of the plantlets were collected from the plants grown in pots in the University campus. They were washed thoroughly in running tap water in order to remove any dirt or soil particles adhered and blotted gently between folds of tissue paper to remove any water droplets. Both enzymic and non-enzymic antioxidants were analysed in all the three parts of the plant. ENZYMIC ANTIOXIDANTS The enzymic antioxidants analyzed in the parts of B. monnieri w ere superoxide dismutase, catalase, peroxidase, glutathione S-transferase and polyphenol oxidase. ASSAY OF SUPEROXIDE DISMUTASE (SOD) SOD was assayed according to the method of Kakkar et al.
    [Show full text]
  • Polyphenol Oxidases in Plants and Fungi: Going Places? a Review
    PHYTOCHEMISTRY Phytochemistry 67 (2006) 2318–2331 www.elsevier.com/locate/phytochem Review Polyphenol oxidases in plants and fungi: Going places? A review Alfred M. Mayer Department of Plant and Environmental Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel Received 3 June 2006; received in revised form 22 July 2006 Available online 14 September 2006 Abstract The more recent reports on polyphenol oxidase in plants and fungi are reviewed. The main aspects considered are the structure, dis- tribution, location and properties of polyphenol oxidase (PPO) as well as newly discovered inhibitors of the enzyme. Particular stress is given to the possible function of the enzyme. The cloning and characterization of a large number of PPOs is surveyed. Although the active site of the enzyme is conserved, the amino acid sequence shows very considerable variability among species. Most plants and fungi PPO have multiple forms of PPO. Expression of the genes coding for the enzyme is tissue specific and also developmentally controlled. Many inhibitors of PPO have been described, which belong to very diverse chemical structures; however, their usefulness for controlling PPO activity remains in doubt. The function of PPO still remains enigmatic. In plants the positive correlation between levels of PPO and the resistance to pathogens and herbivores is frequently observed, but convincing proof of a causal relationship, in most cases, still has not been published. Evidence for the induction of PPO in plants, particularly under conditions of stress and pathogen attack is consid- ered, including the role of jasmonate in the induction process. A clear role of PPO in a least two biosynthetic processes has been clearly demonstrated.
    [Show full text]
  • Discovery of Oxidative Enzymes for Food Engineering. Tyrosinase and Sulfhydryl Oxi- Dase
    Dissertation VTT PUBLICATIONS 763 1,0 0,5 Activity 0,0 2 4 6 8 10 pH Greta Faccio Discovery of oxidative enzymes for food engineering Tyrosinase and sulfhydryl oxidase VTT PUBLICATIONS 763 Discovery of oxidative enzymes for food engineering Tyrosinase and sulfhydryl oxidase Greta Faccio Faculty of Biological and Environmental Sciences Department of Biosciences – Division of Genetics ACADEMIC DISSERTATION University of Helsinki Helsinki, Finland To be presented for public examination with the permission of the Faculty of Biological and Environmental Sciences of the University of Helsinki in Auditorium XII at the University of Helsinki, Main Building, Fabianinkatu 33, on the 31st of May 2011 at 12 o’clock noon. ISBN 978-951-38-7736-1 (soft back ed.) ISSN 1235-0621 (soft back ed.) ISBN 978-951-38-7737-8 (URL: http://www.vtt.fi/publications/index.jsp) ISSN 1455-0849 (URL: http://www.vtt.fi/publications/index.jsp) Copyright © VTT 2011 JULKAISIJA – UTGIVARE – PUBLISHER VTT, Vuorimiehentie 5, PL 1000, 02044 VTT puh. vaihde 020 722 111, faksi 020 722 4374 VTT, Bergsmansvägen 5, PB 1000, 02044 VTT tel. växel 020 722 111, fax 020 722 4374 VTT Technical Research Centre of Finland, Vuorimiehentie 5, P.O. Box 1000, FI-02044 VTT, Finland phone internat. +358 20 722 111, fax + 358 20 722 4374 Edita Prima Oy, Helsinki 2011 2 Greta Faccio. Discovery of oxidative enzymes for food engineering. Tyrosinase and sulfhydryl oxi- dase. Espoo 2011. VTT Publications 763. 101 p. + app. 67 p. Keywords genome mining, heterologous expression, Trichoderma reesei, Aspergillus oryzae, sulfhydryl oxidase, tyrosinase, catechol oxidase, wheat dough, ascorbic acid Abstract Enzymes offer many advantages in industrial processes, such as high specificity, mild treatment conditions and low energy requirements.
    [Show full text]
  • Nelumbo Nucifera)
    Transcriptome Proling for Pericarp Browning During Long-term Storage of Intact Lotus Root (Nelumbo Nucifera) Kanjana Worarad ( [email protected] ) Ibaraki University Tomohiro Suzuki Utsunomiya University Haruka Norii Ibaraki University Yuya Muchizuki Ibaraki University Takashi Ishii Ibaraki Agriculture Center Keiko Shinohara Tokushima Horticulture Takao Miyamoto Renkon3kyodai Tsutomu Kuboyama Ibaraki University Eiichi Inoue Ibaraki University Library Mito Main Library: Ibaraki Daigaku Toshokan Honkan Mito Campus Research Article Keywords: Browning disorder, RNA sequencing, Transcriptomics, Postharvest physiology Posted Date: May 25th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-539513/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Version of Record: A version of this preprint was published at Plant Growth Regulation on August 11th, 2021. See the published version at https://doi.org/10.1007/s10725-021-00736-2. 1 1 Transcriptome profiling for pericarp browning during long-term storage of intact lotus root (Nelumbo 2 nucifera) 3 Kanjana Worarad1, Tomohiro Suzuki2, Haruka Norii1, Yuya Muchizuki1, Takashi Ishii3, Keiko Shinohara4, Takao 4 Miyamoto5, Tsutomu Kuboyama1, Eiichi Inoue1* 5 6 1College of Agriculture, Ibaraki University, Ami, Ibaraki 300-0393, Japan. 7 2Center for Bioscience Research and Education, Utsunomiya University, Utsunomiya, Tochigi 321-8505, 8 Japan. 9 3Ibaraki Agricultural Center, Horticultural Research Institute, Mito, Ibaraki 310-8555, Japan. 10
    [Show full text]
  • Polyphenol Oxidases in Crops: Biochemical, Physiological and Genetic Aspects
    International Journal of Molecular Sciences Review Polyphenol Oxidases in Crops: Biochemical, Physiological and Genetic Aspects Francesca Taranto 1,*, Antonella Pasqualone 2, Giacomo Mangini 2, Pasquale Tripodi 3, Monica Marilena Miazzi 2, Stefano Pavan 2 and Cinzia Montemurro 1,2 1 SINAGRI S.r.l.-Spin off dell’Università degli Studi di Bari “Aldo Moro”, 70126 Bari, Italy; [email protected] 2 Dipartimento di Scienze del Suolo, della Pianta e degli Alimenti, Università degli Studi di Bari “Aldo Moro”, 70126 Bari, Italy; [email protected] (A.P.); [email protected] (G.M.); [email protected] (M.M.M.); [email protected] (S.P.) 3 Consiglio per la ricerca in agricoltura e l’analisi dell’economia agraria, Centro di ricerca per l’orticoltura, 84098 Pontecagnano Faiano, Italy; [email protected] * Correspondence: [email protected]; Tel.: +39-80-5443003 Academic Editor: Gopinadhan Paliyath Received: 15 November 2016; Accepted: 4 February 2017; Published: 10 February 2017 Abstract: Enzymatic browning is a colour reaction occurring in plants, including cereals, fruit and horticultural crops, due to oxidation during postharvest processing and storage. This has a negative impact on the colour, flavour, nutritional properties and shelf life of food products. Browning is usually caused by polyphenol oxidases (PPOs), following cell damage caused by senescence, wounding and the attack of pests and pathogens. Several studies indicated that PPOs play a role in plant immunity, and emerging evidence suggested that PPOs might also be involved in other physiological processes. Genomic investigations ultimately led to the isolation of PPO homologs in several crops, which will be possibly characterized at the functional level in the near future.
    [Show full text]
  • Inhibition of Tyrosinase-Induced Enzymatic Browning by Sulfite and Natural Alternatives
    Inhibition of tyrosinase-induced enzymatic browning by sulfite and natural alternatives Tomas F.M. Kuijpers Thesis committee Promotor Prof. Dr H. Gruppen Professor of Food Chemistry Wageningen University Co-promotor Dr J-P. Vincken Assistant professor, Laboratory of Food Chemistry Wageningen University Other members Prof. Dr M.A.J.S van Boekel, Wageningen University Dr H.T.W.M. van der Hijden, Unilever R&D, Vlaardingen, The Netherlands Dr C.M.G.C. Renard, INRA, Avignon, France Prof. Dr H. Hilz, Hochschule Bremerhaven, Germany This research was conducted under the auspices of the Graduate School VLAG (Advanced studies in Food Technology, Agrobiotechnology, Nutrition and Health Sciences). Inhibition of tyrosinase-induced enzymatic browning by sulfite and natural alternatives Tomas F.M. Kuijpers Thesis submitted in fulfilment of the requirements for the degree of doctor at Wageningen University by the authority of the Rector Magnificus Prof. Dr M.J. Kropff, in the presence of the Thesis Committee appointed by the Academic Board to be defended in public on Friday 25 October 2013 at 4 p.m. in the Aula. Tomas F.M. Kuijpers Inhibition of tyrosinase-mediated enzymatic browning by sulfite and natural alternatives 136 pages PhD thesis, Wageningen University, Wageningen, NL (2013) With references, with summaries in English and Dutch ISBN: 978-94-6173-668-0 ABSTRACT Although sulfite is widely used to counteract enzymatic browning, its mechanism has remained largely unknown. We describe a double inhibitory mechanism of sulfite on enzymatic browning, affecting both the enzymatic oxidation of phenols into o-quinones, as well as the non-enzymatic reactions of these o-quinones into brown pigments.
    [Show full text]
  • A Novel DUF569 Gene Is a Positive Regulator of the Drought Stress Response in Arabidopsis
    International Journal of Molecular Sciences Article A Novel DUF569 Gene Is a Positive Regulator of the Drought Stress Response in Arabidopsis Rizwana Begum Syed Nabi 1,2,†, Rupesh Tayade 1,† , Adil Hussain 3 , Arjun Adhikari 1 , In-Jung Lee 1, Gary J. Loake 4,* and Byung-Wook Yun 1,* 1 School of Applied Biosciences, Kyungpook National University, Daegu 41566, Korea; [email protected] (R.B.S.N.); [email protected] (R.T.); [email protected] (A.A.); [email protected] (I.-J.L.) 2 Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration, Miryang 50424, Korea 3 Department of Agriculture, Abdul Wali Khan University, Mardan 230200, Pakistan; [email protected] 4 Institute of Molecular Plant Sciences, School of Biological Sciences, University of Edinburgh, King’s Buildings, Edinburgh EH9 3JH, UK * Correspondence: [email protected] (G.J.L.); [email protected] (B.-W.Y.) † These authors contributed equally to this work. Abstract: In the last two decades, global environmental change has increased abiotic stress on plants and severely affected crops. For example, drought stress is a serious abiotic stress that rapidly and substantially alters the morphological, physiological, and molecular responses of plants. In Arabidopsis, several drought-responsive genes have been identified; however, the underlying molecular mechanism of drought tolerance in plants remains largely unclear. Here, we report that the Citation: Nabi, R.B.S.; Tayade, R.; “domain of unknown function” novel gene DUF569 (AT1G69890) positively regulates drought stress Hussain, A.; Adhikari, A.; Lee, I.-J.; in Arabidopsis. The Arabidopsis loss-of-function mutant atduf569 showed significant sensitivity Loake, G.J.; Yun, B.-W.
    [Show full text]
  • Open Karma James-Dissertation.Pdf
    The Pennsylvania State University The Graduate School Department of Food Science EFFECT OF DIETARY PRETREATMENT AND OBESITY ON (-)- EPIGALLOCATECHIN-3- GALLATE (EGCG) MEDIATED HEPATOTOXICITY AND THE UNDERLYING MECHANISM A Dissertation in Food Science by Karma James 2016 Karma James Submitted in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy May 2016 The dissertation of Karma James was reviewed and approved* by the following: Joshua Lambert Associate Professor of Food Science Co-Director, Center for Plant and Mushroom Foods for Health Disseration Advisor Chair of Committee Ryan Elias Frederick Sr. and Faith E. Rasmussen Professor of Food Science Associate Professor of Food Science Jairam Vanamala Associate Professor of Food Science Faculty Member, The Penn State Hershey Cancer Institute Faculty Member, Center for Molecular Immunology and Infectious Disease Gary Perdew John T. and Paige S. Smith Professor in Agricultural Sciences Director of the Center for Molecular Toxicology and Carcinogenesis Robert Roberts Professor of Food Science Head of the Department of Food Science *Signatures are on file in the Graduate School iii ABSTRACT A growing body of evidence suggests the hepatotoxic potential of high doses of EGCG. In vitro several mechanisms of EGCG cytotoxicity have been examined; however potential mechanisms of in vivo toxicity remain understudied. Additionally factors that can potentiate EGCG induced hepatotoxicity, such as pre-existing physiological conditions that attenuate liver function have not been investigated. With the lack of data to support an association between dietary supplements and liver toxicity, I hypothesized that dietary pretreatment with EGCG markedly decreases EGCG bioavailability and can potentially mitigate the toxic potential of EGCG, and that ORFLD increases susceptibility to high dose EGCG-mediated hepatotoxicity.
    [Show full text]
  • Chemistry of Secondary Polyphenols Produced During Processing of Tea and Selected Foods
    Int. J. Mol. Sci. 2010, 11, 14-40; doi:10.3390/ijms11010014 OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 www.mdpi.com/journal/ijms Review Chemistry of Secondary Polyphenols Produced during Processing of Tea and Selected Foods Takashi Tanaka *, Yosuke Matsuo and Isao Kouno Laboratory of Natural Product Chemistry, Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan; E-Mails: [email protected] (Y.M.); [email protected] (I.K.) * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +81-95-819-2433; Fax: +81-95-819-2477. Received: 19 November 2009; in revised form: 19 December 2009 / Accepted: 24 December 2009 / Published: 28 December 2009 Abstract: This review will discuss recent progress in the chemistry of secondary polyphenols produced during food processing. The production mechanism of the secondary polyphenols in black tea, whisky, cinnamon, and persimmon fruits will be introduced. In the process of black tea production, tea leaf catechins are enzymatically oxidized to yield a complex mixture of oxidation products, including theaflavins and thearubigins. Despite the importance of the beverage, most of the chemical constituents have not yet been confirmed due to the complexity of the mixture. However, the reaction mechanisms at the initial stages of catechin oxidation are explained by simple quinone–phenol coupling reactions. In vitro model experiments indicated the presence of interesting regio- and stereoselective reactions. Recent results on the reaction mechanisms will be introduced. During the aging of whisky in oak wood barrels, ellagitannins originating from oak wood are oxidized and react with ethanol to give characteristic secondary ellagitannins.
    [Show full text]
  • Inhibition of Oxidative and Antioxidative Enzymes by Trans-Resveratrol X
    JFS: Food Chemistry and Toxicology Inhibition of Oxidative and Antioxidative Enzymes by Trans-Resveratrol X. FAN AND J.P. MATTHEIS Food Chemistry and Toxicology ABSTRACT: Trans-resveratrol, a phytoalexin produced by a variety of plants, has been shown to inhibit oxidative enzymes in an animal cell system. Its effect on several oxidative and antioxidative enzymes from plants was investigated using in vitro assays. Trans-resveratrol inhibited superoxide dismutase, lipoxygenase, catalase, peroxi- dase, polyphenol oxidase, and 1-aminocyclopropane-1-carboxylic acid oxidase with apparent KI’s of 10, 90, 100, 255, 305, and 350 mM, respectively. Trans-resveratrol inhibited lipoxygenase activity more effectively than other lipoxygenase inhibitors, including propyl gallate, ibuprofen, ursolic acid, acetylsalicylic acid, and salicylhydroxamic acid. Key Words: trans-resveratrol, superoxide dismutase, lipoxygenase, catalase, peroxidase, polyphenol oxidase, and 1-aminocyclopropane-1-carboxylic acid oxidase. Introduction environmental stress or pathogenic attack (Langcake and XIDATIVE ENZYMES ARE OF INCREASING INTEREST TO THE Pryce 1976). However, the physiological function of trans- Oplant scientist as well as the food industry because of resveratrol in plants is not well defined. In this study, the ef- their effects on plant metabolism and the nutritional benefits fect of trans-resveratrol on several oxidative and antioxida- they impart to plant foods. Polyphenol oxidase (PPO) is re- tive enzymes found in plants was investigated. sponsible for enzymatic browning in a wide range of vegeta- ble, fruit, and juices (Mayer and Harel 1979). Peroxidases Materials and Methods may be responsible for the generation of free radicals, and may have a role in lignin synthesis, hormone metabolism, Chemicals. and response to stress (Gaspar and others 1982).
    [Show full text]
  • Aravind Jukanti Polyphenol Oxidases (Ppos) in Plants Polyphenol Oxidases (Ppos) in Plants Aravind Jukanti
    Aravind Jukanti Polyphenol Oxidases (PPOs) in Plants Polyphenol Oxidases (PPOs) in Plants Aravind Jukanti Polyphenol Oxidases (PPOs) in Plants Aravind Jukanti Indian Institute of Rice Research Indian Council for Agricultural Research Hyderabad, Telangana, India ISBN 978-981-10-5746-5 ISBN 978-981-10-5747-2 (eBook) DOI 10.1007/978-981-10-5747-2 Library of Congress Control Number: 2017948665 © Springer Nature Singapore Pte Ltd. 2017 This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. The publisher, the authors and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty, express or implied, with respect to the material contained herein or for any errors or omissions that may have been made. The publisher remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Printed on acid-free paper This Springer imprint is published by Springer Nature The registered company is Springer Nature Singapore Pte Ltd.
    [Show full text]