DOCSLIB.ORG

Age-Dependent Absolute Abundance of Hepatic Carboxylesterases

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Age-Dependent Absolute Abundance of Hepatic Carboxylesterases Supplemental material to this article can be found at: http://dmd.aspetjournals.org/content/suppl/2016/11/28/dmd.116.072652.DC1 1521-009X/45/2/216–223$25.00 http://dx.doi.org/10.1124/dmd.116.072652 DRUG METABOLISM AND DISPOSITION Drug Metab Dispos 45:216–223, February 2017 Copyright ª 2017 by The American Society for Pharmacology and Experimental Therapeutics Age-Dependent Absolute Abundance of Hepatic Carboxylesterases (CES1 and CES2) by LC-MS/MS Proteomics: Application to PBPK Modeling of Oseltamivir In Vivo Pharmacokinetics in Infants s Mikael Boberg, Marc Vrana, Aanchal Mehrotra, Robin E. Pearce, Andrea Gaedigk, Deepak Kumar Bhatt, J. Steven Leeder, and Bhagwat Prasad Department of Pharmaceutics, University of Washington, Seattle, Washington (M.B., M.V., A.M., D.K.B., B.P.); Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden (M.B.); Division of Clinical Pharmacology, Toxicology, and Therapeutic Innovation, Children’s Mercy Kansas City, Kansas City, Missouri (R.E.P., A.G., J.S.L.); and School of Medicine, University of Missouri-Kansas City, Kansas City, Missouri (R.E.P., A.G., J.S.L.) Received July 22, 2016; accepted November 22, 2016 Downloaded from ABSTRACT The age-dependent absolute protein abundance of carboxylester- milligram total protein) increased approximately 5-fold (315.2 vs. ase (CES) 1 and CES2 in human liver was investigated and ap- 1664.4) and approximately 3-fold (59.8 vs. 174.1) from neonates to plied to predict infant pharmacokinetics (PK) of oseltamivir. The adults, respectively. CES1 protein abundance in liver cytosol also CES absolute protein abundance was determined by liquid showed age-dependent maturation. Oseltamivir carboxylase activ- dmd.aspetjournals.org chromatography-tandem mass spectrometry proteomics in human ity was correlated with protein abundance in pediatric and adult liver liver microsomal and cytosolic fractions prepared from tissue microsomes. The protein abundance data were then used to model samples obtained from 136 pediatric donors and 35 adult donors. in vivo PK of oseltamivir in infants using pediatric physiologically Two surrogate peptides per protein were selected for the quantifi- based PK modeling and incorporating the protein abundance–based cation of CES1 and CES2 protein abundance. Purified CES1 and ontogeny function into the existing pediatric Simcyp model. The CES2 protein standards were used as calibrators, and the heavy predicted pediatric area under the curve, maximal plasma concen- labeled peptides were used as the internal standards. In he- tration, and time for maximal plasma concentration values were at ASPET Journals on September 25, 2021 patic microsomes, CES1 and CES2 abundance (in picomoles per below 2.1-fold of the clinically observed values, respectively. Introduction or activate drugs (Hines et al., 2013). For example, Gray baby syndrome Because clinical dose optimization studies often have not been resulted in serious adverse effects when the antibiotic chloramphenicol conducted in children, a majority of the drugs used in pediatrics are was given to infants in whom immature glucuronidation resulted in prescribed off-label (Kimland et al., 2012; Laughon et al., 2014). This reduced clearance of the drug (Sutherland, 1959). Therefore, the current suboptimal practice could be unsafe because children, especially approach to predicting pediatric drug dosing, which relies on empirical neonates and infants, may be susceptible to adverse drug effects or lack body weight or body surface area normalization, should also consider the of drug efficacy due to their insufficient ability to metabolically eliminate developmental trajectories of processes involved in drug disposition and response. One solution is to use pediatric physiologically based pharma- cokinetic (pPBPK) models, which can be developed by integrating information on relevant age-dependent physiologic differences, for The study was supported by the Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD) of the National Institutes of Health dose-exposure simulations in children as an alternative to applying scaling [NIH; Grant R01-HD-081299]. The project, entitled “Laboratory of Developmental factors to models based on adult data. In the currently available pPBPK Biology,” was supported by an award from the Eunice Kennedy Shriver NICHD of models, different physiologic factors affecting drug disposition, such as the NIH [Award 5R24-HD-0008836]. Human liver microsomal and cytosolic organ size, tissue composition, pH in the gastrointestinal (GI) tract, and samples from pediatric donors were obtained from the Eunice Kennedy Shriver body fluid dynamics, are taken into consideration (Zhao et al., 2011). NICHD Brain and Tissue Bank for Developmental Disorders at the University of However, a major limitation of existing pPBPK models is the limited Maryland [funded by NIH Contract HHSN275200900011C, reference number availability of precise developmental trajectories for major drug- N01-HD-9-0011]; and from the Liver Tissue Cell Distribution System [funded by metabolizing enzymes (DMEs) and transporters in the drug disposition NIH Contract N01-DK-7-0004/HHSN267200700004C] at the University of Minnesota and the University of Pittsburgh. organs. Therefore, it is critical to study the developmental expression and M.B. and M.V. contributed equally to this work. activity of DMEs and drug transporters (Prasad et al., 2016). dx.doi.org/10.1124/dmd.116.072652. Carboxylesterases (CESs) are DMEs in the a-b hydrolase family, s This article has supplemental material available at dmd.aspetjournals.org. which metabolize many compounds, including drugs containing ester, ABBREVIATIONS: Adultmax, adult maximum average relative protein abundance; AUC, area under the curve; CES, carboxylesterase; CI, confidence interval; CLint, intrinsic clearance; Cmax, maximal plasma concentration; DME, drug-metabolizing enzyme; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MRM, multiple reaction monitoring; m/z, charge/mass ratio; pPBPK, pediatric physiologically based pharmacokinetic; PK, pharmacokinetics; Tmax, time for maximal plasma concentration. 216 Age-Dependent Hepatic Carboxylesterase Abundance 217 thioester, and amide bonds and environmental toxins such as phthalates protein internal standard to address sample-to-sample variability in the trypsin and benzoates. Cocaine (Pindel et al., 1997), heroin (Kamendulis et al., digestion. The mix was then incubated at 95° for 10 minutes with gentle shaking at 1996), clopidogrel (Tang et al., 2006), aspirin (Tang et al., 2006), 300 rpm. Samples were cooled to room temperature for 10 minutes before adding m methylphenidate (Merali et al., 2014), enalapril/ramipril (Thomsen et al., 20 l of iodoacetamide (500 mM) for incubation at room temperature in the dark for m 2014), oseltamivir (Shi et al., 2006), and irinotecan (Haaz et al., 1997) 30 minutes. Five hundred microliters of ice-cold methanol, 100 lofice-cold chloroform, and 400 ml of cold water were added to each sample; vortex mixed; and are some examples of drugs and prodrugs that undergo phase I subjectedtocentrifugation at 16,000g (4°C) for 5 minutes. The upper and lower metabolism by CESs, most of which are prescribed to children. In this layers were removed using vacuum suction, and pellets were dried at room study, hepatic CES1 and CES2 protein abundance in different pediatric temperature for 10 minutes. Pellets were subsequently washed with 500 mlofice- age groups was determined and compared with the observed protein cold methanol and subjected to centrifugation at 8000g (4°C) for 5 minutes, after abundance in adults. Although ontogeny data for hepatic CES1 and which the supernatant was removed. Pellets were dried at room temperature for CES2 have been reported previously (Yang et al., 2009; Hines et al., 30 minutes and resuspended in 60 ml of ammonium bicarbonate buffer (50 mM, pH 2016), we used a state-of-the-art liquid chromatography-tandem mass 7.8). Twenty microliters of trypsin (0.16 mg/ml) was added for digestion (37°C, spectrometry (LC-MS/MS)–based absolute protein quantification ap- 16 hours, gentle shaking at 300 rpm). The trypsin digestion was quenched by proach to quantify age-dependent expression of these proteins in a large placing samples on dry ice. Twenty microliters of heavy peptide internal standard cohort of well-studied pediatric and adult samples. Subsequently, the cocktail [dissolved in acetonitrile/water 80:20 (v/v) with 0.5% formic acid] and 10 ml of acetonitrile/water 80:20 (v/v) with 0.5% formic acid were added to each ontogeny data were incorporated into the Simcyp PBPK software sample to facilitate peptide solubility. After mixing and centrifugation at 4000g (Certara, Princeton, NJ) to predict in vivo pharmacokinetics (PK) of (4°C) for 5 minutes, samples were transferred to LC-MS/MS autosampler vials. Downloaded from oseltamivir in infants. Quantitative Analysis of CESs by LC-MS/MS. The LC-MS/MS system consisted of an Acquity UPLC System (Waters Technologies, Milford, MA) coupled toanABSciexTripleQuad6500System(AppliedBiosystems,Framingham,MA) Materials and Methods was used. Two surrogate peptides per protein were selected for the quantification of Materials. Synthetic heavy labeled peptides (Supplemental Table 1) were CES1 and CES2 protein abundance (Supplemental Table 1) following a previously obtained from Thermo Fisher Scientific (Rockford, IL). Chloroform, ethyl ether, established protocol (Prasad and Unadkat, 2014). Peptide separation was
Load more

Recommended publications
  • Toxicogenomics Applications of New Functional Genomics Technologies in Toxicology
    \-\w j Toxicogenomics Applications of new functional genomics technologies in toxicology Wilbert H.M. Heijne Proefschrift ter verkrijging vand egraa dva n doctor opgeza gva nd e rector magnificus vanWageninge n Universiteit, Prof.dr.ir. L. Speelman, in netopenbaa r te verdedigen op maandag6 decembe r200 4 des namiddagst e half twee ind eAul a - Table of contents Abstract Chapter I. page 1 General introduction [1] Chapter II page 21 Toxicogenomics of bromobenzene hepatotoxicity: a combined transcriptomics and proteomics approach[2] Chapter III page 48 Bromobenzene-induced hepatotoxicity atth etranscriptom e level PI Chapter IV page 67 Profiles of metabolites and gene expression in rats with chemically induced hepatic necrosis[4] Chapter V page 88 Liver gene expression profiles in relation to subacute toxicity in rats exposed to benzene[5] Chapter VI page 115 Toxicogenomics analysis of liver gene expression in relation to subacute toxicity in rats exposed totrichloroethylen e [6] Chapter VII page 135 Toxicogenomics analysis ofjoin t effects of benzene and trichloroethylene mixtures in rats m Chapter VII page 159 Discussion and conclusions References page 171 Appendices page 187 Samenvatting page 199 Dankwoord About the author Glossary Abbreviations List of genes Chapter I General introduction Parts of this introduction were publishedin : Molecular Biology in Medicinal Chemistry, Heijne etal., 2003 m NATO Advanced Research Workshop proceedings, Heijne eral., 2003 81 Chapter I 1. General introduction 1.1 Background /.1.1 Toxicologicalrisk
    [Show full text]
  • Baboon JMP CES Paper
    Baboon carboxylesterases 1 and 2: sequences, structures and phylogenetic relationships with human and other primate carboxylesterases Author Holmes, Roger S, Glenn, Jeremy P, VandeBerg, John L, Cox, Laura A Published 2009 Journal Title Journal of Medical Primatology DOI https://doi.org/10.1111/j.1600-0684.2008.00315.x Copyright Statement © 2009 John Wiley & Sons A/S. This is the author-manuscript version of the paper. Reproduced in accordance with the copyright policy of the publisher.The definitive version is available at www.interscience.wiley.com Downloaded from http://hdl.handle.net/10072/29362 Griffith Research Online https://research-repository.griffith.edu.au Baboon Carboxylesterases 1 and 2: Sequences, Structures and Phylogenetic Relationships with Human and other Primate Carboxylesterases Roger S. Holmes 1,2,3 , Jeremy P. Glenn 1, John L. VandeBerg 1,2 , Laura A. Cox 1,2,4 1. Department of Genetics and 2. Southwest National Primate Research Center, Southwest Foundation for Biomedical Research, San Antonio, TX, USA, and 3. School of Biomolecular and Physical Sciences, Griffith University, Nathan. Queensland, Australia 4. Corresponding Author: Laura A. Cox, Ph.D. Department of Genetics Southwest National Primate Research Center Southwest Foundation for Biomedical Research San Antonio, TX, USA 78227 Email: [email protected] Phone: 210-258-9687 Fax: 210-258-9600 Keywords: cDNA sequence; amino acid sequence; 3-D structure Running Head: Carboxylesterases: sequences and phylogeny Published in Journal of Medical Primatology (2009) 38: 27-38. Abstract Background Carboxylesterase (CES) is predominantly responsible for the detoxification of a wide range of drugs and narcotics, and catalyze several reactions in cholesterol and fatty acid metabolism.
    [Show full text]
  • NIH Public Access Author Manuscript Mamm Genome
    NIH Public Access Author Manuscript Mamm Genome. Author manuscript; available in PMC 2011 October 8. NIH-PA Author ManuscriptPublished NIH-PA Author Manuscript in final edited NIH-PA Author Manuscript form as: Mamm Genome. 2010 October ; 21(9-10): 427±441. doi:10.1007/s00335-010-9284-4. Recommended nomenclature for five mammalian carboxylesterase gene families: human, mouse, and rat genes and proteins Roger S. Holmes Department of Genetics, Southwest Foundation for Biomedical Research, San Antonio, TX 78227-5301, USA Southwest National Primate Research Center, Southwest Foundation for Biomedical Research, San Antonio, TX, USA School of Biomolecular and Physical Sciences, Griffith University, Brisbane, QLD, Australia Matthew W. Wright European Bioinformatics Institute, Wellcome Trust Genome Campus, Cambridge, UK Stanley J. F. Laulederkind Rat Genome Database, Human Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, WI, USA Laura A. Cox Department of Genetics, Southwest Foundation for Biomedical Research, San Antonio, TX 78227-5301, USA Southwest National Primate Research Center, Southwest Foundation for Biomedical Research, San Antonio, TX, USA Masakiyo Hosokawa Laboratory of Drug Metabolism and Biopharmaceutics, Chiba Institute of Science, Choshi, Chiba, Japan Teruko Imai Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto, Japan Shun Ishibashi Department of Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan Richard Lehner CIHR Group on Molecular and Cell Biology of Lipids, University of Alberta, Edmonton, AB, Canada Masao Miyazaki The Institute of Glycoscience, Tokai University, Kanagawa, Japan Everett J. Perkins Department of Drug Disposition, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN, USA Phillip M. Potter © Springer Science+Business Media, LLC 2010 [email protected] . Holmes et al.
    [Show full text]
  • Carboxylesterases in Lipid Metabolism: from Mouse to Human
    Protein Cell DOI 10.1007/s13238-017-0437-z Protein & Cell REVIEW Carboxylesterases in lipid metabolism: from mouse to human & Jihong Lian1,2 , Randal Nelson1,2, Richard Lehner1,2,3 1 Group on Molecular and Cell Biology of Lipids, University of Alberta, Edmonton, Alberta, Canada 2 Department of Pediatrics, University of Alberta, Edmonton, Alberta, Canada 3 Department of Cell Biology, University of Alberta, Edmonton, Alberta, Canada & Correspondence: [email protected] (J. Lian) Received March 2, 2017 Accepted May 31, 2017 Cell & ABSTRACT Hatfield et al., 2016; Fukami et al., 2015; Laizure et al., 2013; Staudinger et al., 2010; Sanghani et al., 2009; Imai, 2006). Mammalian carboxylesterases hydrolyze a wide range However, carboxylesterases have also been demonstrated of xenobiotic and endogenous compounds, including to hydrolyze endogenous esters and thioesters including lipid esters. Physiological functions of car- lipids and some of these enzymes have been shown to play Protein boxylesterases in lipid metabolism and energy home- important physiological functions in lipid metabolism and ostasis in vivo have been demonstrated by genetic energy homeostasis. Recent research endeavors have manipulations and chemical inhibition in mice, and provided more insight into the roles of human car- in vitro through (over)expression, knockdown of boxylesterases in metabolic diseases. expression, and chemical inhibition in a variety of cells. Genes encoding six human carboxylesterases and twenty Recent research advances have revealed the relevance mouse carboxylesterases have been classified. However, of carboxylesterases to metabolic diseases such as given the interspecies diversity of carboxylesterases both in obesity and fatty liver disease, suggesting these the number and primary amino acid sequences there is a enzymes might be potential targets for treatment of need to define functional mouse and human orthologs.
    [Show full text]
  • Identification and Characterization of Carboxylesterase Variants
    Identification and characterization of Carboxylesterase variants Master thesis for the degree Master of Pharmacy Kathleen Nanding School of Pharmacy The Faculty of Mathematics and Natural Sciences UNIVERSITY OF OSLO Department of Pharmacology OSLO UNIVERSITY HOSPITAL, ULLEVÅL April, 2016 I II III IV Copyright © Kathleen Nanding 2016 Identification and characterization of carboxylesterase variants Kathleen Nanding http://www.duo.uio.no Printed Reprosentralen, University of Oslo V VI Preface This thesis is made as a completion of the master education in Pharmacy and it took place at the Department of Pharmacology, Oslo University Hospital, Ullevål during the period of August 2015 to April 2016. First of all, I would like to express my deep gratitude to my external supervisors Marianne K. Kringen and Kari Bente Foss Haug for all the guidance and support through the entire study. I have learnt so much and I am truely insipired. I would also like to thank Hege Gilbø Bakke for helping me with all the practical problems I had, and for her kindness and patience. Furthermore I would like to thank all the colleagues at the department for their help and for a delightful and warm working atomsphere. I am grateful to my internal supervisor Hege Thoresen for following the progress of my study and for her constructive comments on the thesis. My thanks also go to the Class 2016 and my friends for all the support and for joyful memories through the years of my education. Finally I would like to thank my parents for all the love and encouragement, I would have never completed my education without your support.
    [Show full text]
  • Screening of Key Genes Associated with R‑CHOP Immunochemotherapy and Construction of a Prognostic Risk Model in Diffuse Large B‑Cell Lymphoma
    MOLECULAR MEDICINE REPORTS 20: 3679-3690, 2019 Screening of key genes associated with R‑CHOP immunochemotherapy and construction of a prognostic risk model in diffuse large B‑cell lymphoma RAN LIU1*, ZHI CHEN2*, SHUJUN WANG3, GANG ZHAO1, YAN GU1, QI HAN1 and BAOAN CHEN1 1Department of Hematology and Oncology, Zhongda Hospital Southeast University; 2Department of Cardiology, Jiangsu Province Hospital; 3Department of Blood Transfusion, Nanjing General Hospital of PLA, Nanjing, Jiangsu 210009, P.R. China Received February 18, 2019; Accepted July 25, 2019 DOI: 10.3892/mmr.2019.10627 Abstract. Diffuse large B-cell lymphoma (DLBCL) is a enriched in functions associated with ‘response to DNA common subtype of non-Hodgkin lymphoma, which is damage stimulus’, and pathways including ‘cytokine-cytokine curable in the majority of patients treated with rituximab receptor interaction’ and ‘cell cycle’. The optimal combination plus cyclophosphamide, doxorubicin, vincristine and pred- of the 12 genes, including CAMK1, HPCAL4 and EFNA5, was nisone (R‑CHOP) immunochemotherapy. However, the found to be useful in predicting the prognosis of patients with therapeutic mechanism of R‑CHOP has not been elucidated. DLBCL after R‑CHOP treatment. Therefore, these genes may The GSE32918 and GSE57611 datasets were retrieved from be affected by R‑CHOP in DLBCL. The Gene Expression Omnibus database. The differentially expressed genes (DEGs) associated with R‑CHOP therapy were Introduction identified using limma. Combined with prognostic informa- tion in GSE32918, DEGs found to be significantly associated Diffuse large B-cell lymphoma (DLBCL) is a B-cell malig- with prognosis were selected using univariate Cox regression nancy. B-cells are a type of white blood cell responsible for analysis and a risk prediction model was constructed.
    [Show full text]
  • De Novo Transcriptome Assembly and Annotation of Liver and Brain
    G C A T T A C G G C A T genes Communication De Novo Transcriptome Assembly and Annotation of Liver and Brain Tissues of Common Brushtail Possums (Trichosurus vulpecula) in New Zealand: Transcriptome Diversity after Decades of Population Control Arsalan Emami-Khoyi 1,2 , Shilpa Pradeep Parbhu 1 , James G. Ross 2 , Elaine C. Murphy 2, Jennifer Bothwell 2 , Daniela M. Monsanto 1 , Bettine Jansen van Vuuren 1 , Peter R. Teske 1 and Adrian M. Paterson 2,* 1 Center for Ecological Genomics and Wildlife Conservation, University of Johannesburg, Auckland Park 2006, South Africa; [email protected] (A.E.-K.); [email protected] (S.P.P.); [email protected] (D.M.M.); [email protected] (B.J.v.V.); [email protected] (P.R.T.) 2 Department of Pest-management and Conservation, Faculty of Agriculture and Life Sciences, Lincoln University, Lincoln 7647, New Zealand; [email protected] (J.G.R.); [email protected] (E.C.M.); [email protected] (J.B.) * Correspondence: [email protected] Received: 27 February 2020; Accepted: 10 April 2020; Published: 17 April 2020 Abstract: The common brushtail possum (Trichosurus vulpecula), introduced from Australia in the mid-nineteenth century, is an invasive species in New Zealand where it is widespread and forms the largest self-sustained reservoir of bovine tuberculosis (Mycobacterium bovis) among wild populations. Conservation and agricultural authorities regularly apply a series of population control measures to suppress brushtail possum populations. The evolutionary consequence of more than half a century of intensive population control operations on the species’ genomic diversity and population structure is hindered by a paucity of available genomic resources.
    [Show full text]
  • Report on Research and Other Activities of National Institute of Genetics (During 2014~2018)
    Report on Research and Other Activities of National Institute of Genetics (During 2014~2018) ᅜ❧㑇ఏᏛ◊✲ᡤ⮬ᕫホ౯ሗ࿌᭩ National Institute of Genetics October 2019 ᅜ❧㑇ఏᏛ◊✲ᡤ 2019 ᖺ 10 ᭶ CONTENTS 1. Preface 2. Overview of National Institute of Genetics 2 z Our Mission 2 z History 3 z Organization 5 z Outline of Departments and Centers for Research Infrastructures 6 3. Research Achievements and Self Evaluation 7 z Department of Informatics 7 z Department of Genomics and Evolutionary Biology 10 z Department of Gene Function and Phenomics 13 z Department of Chromosome Science 18 z Center for Frontier Research 21 4. Intellectual Infrastructure Center 22 z Bioinformation and DDBJ Center 22 z Advanced Genomics Center 29 z Genetic Resource Center 32 5. Joint Research and Research Meeting (NIG-JOINT), and International Symposium 41 6. Education and Outreach 42 z Graduate Education 42 z Education: Outreach 47 7. Activities to Promote International Interaction and Cooperation 51 8. Office for Research Development 53 9. NIG INNOVATION 55 10. Office for Gender Equality56 11. Publication Lists 57 Preface Director-General: Fumio Hanaoka The National Institute of Genetics (NIG) recently celebrated its 70th anniversary since its founding and continues to strive for excellence in research, service and education. To pursue our commitment to future development, we have initiated an evaluation by an external review committee and have compiled this report which summarizes the activity of NIG during the 5 years from January 2014 to December 2018. NIG was established in 1949 by the Japanese Ministry of Education for comprehensive studies of genetics. In 1984, it was reorganized into an Inter-University Research Institute, and the new designation established an additional official role of NIG: collaboration with researchers at universities.
    [Show full text]
  • Expression of the Human Carboxylesterase 2 Enzyme (CES2) in Mammalian Cells
    Bárbara Martins Paiva da Cunha Degree in Biochemistry Expression of the human carboxylesterase 2 enzyme (CES2) in mammalian cells Dissertation to obtain a Master Degree in Biotechnology Supervisor: Ana Luísa Simplício, Ph.D, IBET/ITQB-UNL Co-Supervisor: Ana Sofia Coroadinha, Ph.D, IBET/ITQB-UNL Júri: Presidente: Prof. Doutor Rui Manuel Freitas de Oliveira Arguente: Profª. Doutora Alexandra Fernandes Vogal: Doutora Ana Luísa Ferreira Simplício September 2011 II Bárbara Martins Paiva da Cunha Degree in Biochemistry Expression of the human carboxylesterase 2 enzyme (CES2) in mammalian cells Dissertation to obtain a Master Degree in Biotechnology Supervisor: Ana Luísa Simplício, Ph.D, IBET/ITQB-UNL Co-Supervisor: Ana Sofia Coroadinha, Ph.D, IBET/ITQB-UNL Júri: Presidente: Prof. Doutor Rui Manuel Freitas de Oliveira Arguente: Profª. Doutora Alexandra Fernandes Vogal: Ana Luísa Ferreira Simplício September 2011 IV Copyright "Expression of the human carboxylesterase 2 enzyme (CES2) in mammalian cells". Copyright Bárbara Martins Paiva da Cunha, FCT/UNL, UNL A Faculdade de Ciências e Tecnologia e a Universidade Nova de Lisboa têm o direito, perpétuo e sem limites geográficos, de arquivar e publicar esta dissertação através de exemplares impressos reproduzidos em papel ou de forma digital, ou por qualquer outro meio conhecido ou que venha a ser inventado, e de a divulgar através de repositórios científicos e de admitir a sua cópia e distribuição com objectivos educacionais ou de investigação, não comerciais, desde que seja dado crédito ao autor e editor. V VI Acknowledgments Acknowledgments Gostaria de agradecer a todas as pessoas que directa ou indirectamente contribuíram para a realização deste trabalho e sem as quais este não teria sido possível.
    [Show full text]
  • 1 Imipramine Treatment and Resiliency Exhibit Similar
    Imipramine Treatment and Resiliency Exhibit Similar Chromatin Regulation in the Mouse Nucleus Accumbens in Depression Models Wilkinson et al. Supplemental Material 1. Supplemental Methods 2. Supplemental References for Tables 3. Supplemental Tables S1 – S24 SUPPLEMENTAL TABLE S1: Genes Demonstrating Increased Repressive DimethylK9/K27-H3 Methylation in the Social Defeat Model (p<0.001) SUPPLEMENTAL TABLE S2: Genes Demonstrating Decreased Repressive DimethylK9/K27-H3 Methylation in the Social Defeat Model (p<0.001) SUPPLEMENTAL TABLE S3: Genes Demonstrating Increased Repressive DimethylK9/K27-H3 Methylation in the Social Isolation Model (p<0.001) SUPPLEMENTAL TABLE S4: Genes Demonstrating Decreased Repressive DimethylK9/K27-H3 Methylation in the Social Isolation Model (p<0.001) SUPPLEMENTAL TABLE S5: Genes Demonstrating Common Altered Repressive DimethylK9/K27-H3 Methylation in the Social Defeat and Social Isolation Models (p<0.001) SUPPLEMENTAL TABLE S6: Genes Demonstrating Increased Repressive DimethylK9/K27-H3 Methylation in the Social Defeat and Social Isolation Models (p<0.001) SUPPLEMENTAL TABLE S7: Genes Demonstrating Decreased Repressive DimethylK9/K27-H3 Methylation in the Social Defeat and Social Isolation Models (p<0.001) SUPPLEMENTAL TABLE S8: Genes Demonstrating Increased Phospho-CREB Binding in the Social Defeat Model (p<0.001) SUPPLEMENTAL TABLE S9: Genes Demonstrating Decreased Phospho-CREB Binding in the Social Defeat Model (p<0.001) SUPPLEMENTAL TABLE S10: Genes Demonstrating Increased Phospho-CREB Binding in the Social
    [Show full text]
  • Human Carboxylesterase 2 Splice Variants
    HUMAN CARBOXYLESTERASE 2 SPLICE VARIANTS: EXPRESSION, ACTIVITY, AND ROLE IN THE METABOLISM OF IRINOTECAN AND CAPECITABINE Marissa Ann Schiel Submitted to the faculty of the University Graduate School in partial fulfillment of the requirements for the degree Doctor of Philosophy in the Department of Biochemistry and Molecular Biology, Indiana University February 2009 Accepted by the Faculty of Indiana University, in partial fulfillment of the requirements for the degree of Doctor of Philosophy. William F. Bosron, Ph.D., Chair E. Gabriela Chiorean, M.D. Doctoral Committee David A. Flockhart, M.D., Ph.D. Maureen A. Harrington, Ph.D. August 28, 2008 Sonal P. Sanghani, Ph.D. ii To my Poppa, Who encouraged me to finish and do my best. iii ACKNOWLEDGEMENTS I am sincerely thankful for all the help and encouragement I have received while pursuing my graduate education. I would like to gratefully acknowledge the following individuals: Dr. William Bosron for his passion for both science and education. I met Dr. Bosron on my very first visit to IUSM, and I was beyond pleased when I found a place in his lab. His wisdom and generosity truly enhanced my graduate experience. Dr. Sonal Sanghani for her knowledge and guidance during every day of this journey. I am grateful that she is both my mentor and my friend. Dr. Maureen Harrington, committee member and co-director of the MSTP. Her mentorship has been invaluable as I pursued both my graduate and medical studies. With her guidance, I happily did a rotation and found a home in the Bosron lab. She and Dr.
    [Show full text]
  • Age-Dependent Absolute Abundance of Hepatic Carboxylesterases (CES1 And
    DMD Fast Forward. Published on November 28, 2016 as DOI: 10.1124/dmd.116.072652 This article has not been copyedited and formatted. The final version may differ from this version. DMD # 72652 Age-dependent absolute abundance of hepatic carboxylesterases (CES1 and CES2) by LC-MS/MS proteomics: Application to PBPK modeling of oseltamivir in vivo pharmacokinetics in infants Mikael Boberg, Marc Vrana, Aanchal Mehrotra, Robin E. Pearce, Andrea Gaedigk, Deepak Kumar Bhatt, J. Steven Leeder and Bhagwat Prasad Department of Pharmaceutics, University of Washington, Seattle, WA (M.B., M.V., A.M., Downloaded from D.K.B. and B.P.) Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden (M.B.) dmd.aspetjournals.org Department of Clinical Pharmacology, Toxicology & Therapeutic Innovation, Children’s Mercy-Kansas City, MO and School of Medicine, University of Missouri-Kansas City, Kansas City, MO (R.E.P., A.G., J.S.L.) at ASPET Journals on September 28, 2021 1 DMD Fast Forward. Published on November 28, 2016 as DOI: 10.1124/dmd.116.072652 This article has not been copyedited and formatted. The final version may differ from this version. DMD # 72652 Running title: Age-dependent hepatic carboxylesterase abundance Corresponding author: Bhagwat Prasad, Ph.D.; Department of Pharmaceutics, University of Washington, Seattle, WA 98195, Phone: (206) 221-2295, Fax: (206) 543-3204; E-mail: [email protected] Number of text pages: 15 Number of tables: 3 Downloaded from Number of figures: 6 Number of references: 39 Number of words in Abstract: 197 (250) dmd.aspetjournals.org Number of words in Introduction: 449 (750) Number of words in Discussion: 951 (1,500) at ASPET Journals on September 28, 2021 Abbreviations: Area under curve (AUC), carboxylesterase (CES), drug-metabolizing enzyme (DME), maximal plasma concentration (Cmax), pediatric physiologically based pharmacokinetic (pPBPK), pharmacokinetics (PK), multiple reaction monitoring (MRM), liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS), time for maximal plasma concentration (Tmax) 2 DMD Fast Forward.
    [Show full text]