DOCSLIB.ORG

Ph.D Thesis – L. Shaya; Mcmaster University – Department of Biology

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Ph.D Thesis – L. Shaya; Mcmaster University – Department of Biology Ph.D Thesis – L. Shaya; McMaster University – Department of Biology FUNCTION AND REGULATION OF FISH CYP3 GENES Ph.D Thesis – L. Shaya; McMaster University – Department of Biology CHARACTERIZING THE FUNCTION AND REGULATION OF ORPHAN CYP3 GENES IN ZEBRAFISH (DANIO RERIO) By LANA SHAYA, H.B.Sc., M.Sc. A Thesis Submitted to the School of Graduate Studies in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy McMaster University © Copyright by Lana Shaya, August 2019 Ph.D Thesis – L. Shaya; McMaster University – Department of Biology McMaster University DOCTOR OF PHILOSOPHY (2019) Hamilton, Ontario (Biology) TITLE: Characterizing the Function and Regulation of Orphan CYP3C Genes in Zebrafish (Danio rerio) AUTHOR: Lana Shaya, Hon B.Sc. (University of Toronto Mississauga); M.Sc. (McMaster University) SUPERVISOR: Dr. Joanna Y. Wilson NUMBER OF PAGES: xxi, 235 ii Ph.D Thesis – L. Shaya; McMaster University – Department of Biology LAY ABSTRACT Cytochrome P450 (CYP) enzymes break down compounds such as hormones and pharmaceuticals. While mammals have genes in the CYP3A subfamily, fish have unique subfamilies not found in mammals. The function and regulation of the CYP3 family in fish is unknown, but commonly assumed to be like human CYP3. The purpose of this thesis was to identify what receptors and compounds regulate CYP3C enzymes in zebrafish. We found that regulation of CYP3C enzymes in zebrafish is different than humans. Zebrafish CYP3C genes are regulated by the aryl hydrocarbon receptor and estrogen receptor, while human CYP3A is regulated by the pregnane-x-receptor. I used a high throughput approach to screen thousands of compounds to identify the function of CYP3A65 and CYP3C1 from zebrafish. CYP3A65 and CYP3C1 metabolize several plant-based and pharmaceutical compounds. CYP3A65 and CYP3C1 are more functionally similar to each other than to CYP3A in humans. iii Ph.D Thesis – L. Shaya; McMaster University – Department of Biology ABSTRACT Genome sequencing has resulted in the identification of >55,000 cytochrome P450 enzymes, many of which have an unknown function and regulation. In mammals, CYP3 genes appear in only one subfamily (CYP3A), which metabolize >50% of pharmaceuticals and some steroids in humans. Unlike mammals, fish contain genes in the CYP3A, CYP3B, CYP3C and CYP3D subfamilies. While it is commonly assumed that fish and mammalian CYP3A are functional similar, the function and regulation of fish CYP3 remains largely unknown. In this thesis, the receptors and compounds that regulate CYP3C genes in zebrafish were assessed. The induction of CYP3C genes in response to the aryl hydrocarbon (AHR) and estrogen receptor (ER) ligands, β-naphthoflavone and 17β-estradiol, was measured using quantitative PCR in intestine, liver and gonads. Zebrafish CYP3C genes were inducible by β-naphthoflavone and 17β-estradiol, implicating the aryl hydrocarbon and estrogen receptor in CYP3C gene regulation and suggesting that regulation of CYP3 genes in fish differs from that in mammals. To define the function of zebrafish CYP3A65 and CYP3C1, fluorogenic compounds which are specific markers of CYP1 and CYP3A activity in humans, were screened for metabolism by CYP3A65 and CYP3C1. Both CYP3A65 and CYP3C1 had the capacity to metabolize several of these compounds and the substrate profile overlapped with zebrafish CYP1A, suggesting that these compounds are not specific in fish. A high throughput approach was employed to screen ~4000 small biologically and pharmacologically active compounds for metabolism by CYP3A65 and CYP3C1, using NADPH consumption to assess catalytic activity. The substrate profiles of CYP3A65 and CYP3C1 largely overlapped iv Ph.D Thesis – L. Shaya; McMaster University – Department of Biology and were different than mammalian CYP3A4. CYP3A65 and CYP3C1 appeared to have a bias for quinone-based compounds but further studies are required to confirm quinones as substrates and to assess a strong structure-activity relationship. Overall, this study provides insight on the regulation, function and evolution on CYP3 genes in fish. v Ph.D Thesis – L. Shaya; McMaster University – Department of Biology ACKNOWLEDGEMENT During graduate school, I was blessed with great mentorship, supportive colleagues, friends and family, making my experience one I will never forget. I offer sincere thanks to the people who provided me with everything I needed to succeed! Joanna is EVERYTHING a mentor should be. I started in her lab as a shy, know- nothing student who shutdown every time things were not easy. Thanks to her mentorship, I am a strong, confident scientist who can do anything. Joanna, your patience is unmatched! I will never forget the times you willingly made yourself available to listen to a practice presentation for a millionth time or address an unwarranted, panic-inducing science concern. Thank you for taking the time to get to know me as a scientist and a person, and to always selflessly push me towards the path that would get me to where I wanted to be. Thank you for being an amazing mentor and my strongest advocate. I could not have done this with any other supervisor. You will forever be my lab family. At the front and center of my success are my amazing and loving parents, Aadl and Walaa Shaya Gulla. Mom and Dad, thank you for the financial and emotional support you always so willingly provided. There had been countless nights when I felt I could not continue but I took comfort in knowing that you believed in me when I did not believe in myself. No matter how tired you were, you always tried to learn about what I did (even when I was terrible at explaining), and continued to ask for updates throughout the years, and motivated me to work hard. Sometimes the stress of work would cause me to be a less than ideal daughter to ideal parents, and for that I apologize, but thank you for loving and supporting me regardless. I hope that this makes you proud, and that you feel I did justice to your sacrifices for and investments in me. I love you guys so much and I am so lucky to have you in my corner. I am invincible with you by my side. Of course, I must acknowledge the support of my sister and 2 little brothers. They are here because I appreciate their patience during the times I yelled at them when I was PhDlife-level-stressed. To my sister, thank you for being just as PhDlife-level-stressed, so I always had someone at home who got it (and who sometimes was even more stressed and could yell louder than me, making me look like an angel to mom and dad ;) ). To my baby brothers, thanks for taking advantage of my no-time-for-anything-PhDlife and washing my car for an exuberant monetary value; it was worth it! I enjoyed having a spotless looking car. Love you guys. Xo. I would like to thank my fiancé, Michal Galus. The last few months have been the most challenging of my life and I know it made me a challenging person to love. Who decides to buy a house, get a job, write a Ph.D. and plan a wedding all at once? Crazy people, that’s who! Michal, I could not imagine making it through all this craziness without you. Amidst our massive list of responsibilities, you let me prioritize my degree vi Ph.D Thesis – L. Shaya; McMaster University – Department of Biology and career, and stepped up to handle everything that I could not. You encouraged me to go after what I wanted and put my needs above yours. The good news is, now that the thesis is done, there will not be anymore unprovoked anger towards you for having free time when I did not, but the bad news is I have free time now to bother you during yours. A huge thanks to all the past and present members of the Wilson Lab who have provided their technical and emotional support to this project. A special thank you to Shumaila Fraz and Derek Alsop for always being so willing when I needed experimental advice and support. To James and Andrea, for keeping me sane and making the last couple of years of my Ph.D. some of the most enjoyable. Thank you especially for all the anxiety-attack-panic-calming compression hugs that were better than any coping mechanism ever described and for the late nights spent figuring out plasmid orientations or arguing about tip racking. Truly, I could not imagine a better duo to contemplate why on earth my gene construct would not do what it did two days ago. Thank you to my committee members, Ben Evans and Bhagwati Gupta, for always being available to read my drafts and answer any questions. It has been a pleasure learning from you. Thank you to the administrative staff at Biology whom have worked tirelessly to ensure the success of their students. To name a few that have made me life so much easier at Mac: Barb Reuter, Kathy Greaves and Karen Haines. The passion with which you do your job is one I can only aspire to have. Thank you for not making me feel like another number in the system. These are only a few of the many people that had an impact on me during my graduate life, and there is not enough room here to name all the others whom I appreciate and will miss immensely. I am very sad that this chapter of my life has ended but I will forever remember my time at McMaster University and the wonderful people that shaped my experience. vii Ph.D Thesis – L. Shaya; McMaster University – Department of Biology TABLE OF CONTENTS LAY ABSTRACT ............................................................................................................. iii ABSTRACT ........................................................................................................................ iv ACKNOWLEDGEMENT .................................................................................................. vi TABLE OF CONTENTS ................................................................................................
Load more

Recommended publications
  • (12) United States Patent (10) Patent N0.: US 8,343,962 B2 Kisak Et Al
    US008343962B2 (12) United States Patent (10) Patent N0.: US 8,343,962 B2 Kisak et al. (45) Date of Patent: *Jan. 1, 2013 (54) TOPICAL FORMULATION (58) Field of Classi?cation Search ............. .. 514/226.5, 514/334, 420, 557, 567 (75) Inventors: Edward T. Kisak, San Diego, CA (US); See application ?le fOr Complete Search history. John M. NeWsam, La Jolla, CA (US); _ Dominic King-Smith, San Diego, CA (56) References C‘ted (US); Pankaj Karande, Troy, NY (US); Samir Mitragotri, Goleta, CA (US) US' PATENT DOCUMENTS 5,602,183 A 2/1997 Martin et al. (73) Assignee: NuvoResearchOntano (CA) Inc., Mississagua, 6,328,979 2B1 12/2001 Yamashita et a1. 7,001,592 B1 2/2006 Traynor et a1. ( * ) Notice: Subject to any disclaimer, the term of this 7,795,309 B2 9/2010 Kisak eta1~ patent is extended or adjusted under 35 2002/0064524 A1 5/2002 Cevc U.S.C. 154(b) by 212 days. FOREIGN PATENT DOCUMENTS This patent is subject to a terminal dis- W0 WO 2005/009510 2/2005 claimer- OTHER PUBLICATIONS (21) APPI' NO‘, 12/848,792 International Search Report issued on Aug. 8, 2008 in application No. PCT/lB2007/0l983 (corresponding to US 7,795,309). _ Notice ofAlloWance issued on Apr. 29, 2010 by the Examiner in US. (22) Med Aug- 2’ 2010 Appl. No. 12/281,561 (US 7,795,309). _ _ _ Of?ce Action issued on Dec. 30, 2009 by the Examiner in US. Appl. (65) Prior Publication Data No, 12/281,561 (Us 7,795,309), Us 2011/0028460 A1 Feb‘ 3’ 2011 Primary Examiner * Raymond Henley, 111 Related U 5 Application Data (74) Attorney, Agent, or Firm * Foley & Lardner LLP (63) Continuation-in-part of application No.
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 9,393,221 B2 W (45) Date of Patent: Jul.19, 2016
    USOO9393221B2 (12) United States Patent (10) Patent No.: US 9,393,221 B2 W (45) Date of Patent: Jul.19, 2016 (54) METHODS AND COMPOUNDS FOR FOREIGN PATENT DOCUMENTS REDUCING INTRACELLULAR LIPID STORAGE WO WO2007096,251 8, 2007 OTHER PUBLICATIONS (75) Inventor: Sean Wu, Brookline, MA (US) Onyesom and Agho, Asian J. Sci. Res., Oct. 2010, vol. 4, No. 1, p. (73) Assignee: THE GENERAL, HOSPITAL 78-83. CORPORATION, Boston, MA (US) Davis et al., Br J Clin Pharmacol., 1996, vol. 4, p. 415-421.* Schweiger et al., Am J Physiol Endocrinol Metab, 2009, vol. 279, E289-E296. (*) Notice: Subject to any disclaimer, the term of this Maryam Ahmadian et al., Desnutrin/ATGL is regulated by AMPK patent is extended or adjusted under 35 and is required for a brown adipose phenotype, Cell Metabolism, vol. U.S.C. 154(b) by 748 days. 13, pp. 739-748, 2011. Mohammadreza Bozorgmanesh et al., Diabetes prediction, lipid (21) Appl. No.: 13/552,975 accumulation product, and adiposity measures; 6-year follow-up: Tehran lipid and glucose study, Lipids in Health and Disease, vol. 9, (22) Filed: Jul.19, 2012 pp. 1-9, 2010. Judith Fischer et al., The gene encoding adipose triglyceride lipase (65) Prior Publication Data (PNPLA2) is mutated in neutral lipid storage disease with myopathy, Nature Genetics, vol.39, pp. 28-30, 2007. US 2013/OO23488A1 Jan. 24, 2013 Astrid Gruber et al., The N-terminal region of comparative gene identification-58 (CGI-58) is important for lipid droplet binding and activation of adipose triglyceride lipase, vol. 285, pp. 12289-12298, Related U.S.
    [Show full text]
  • (UHA) Formulary and Its Appendices Is Provided for the Convenience of Medical Providers and UHA Members
    INTRODUCTION The information contained in the Umpqua Health Alliance (UHA) Formulary and its appendices is provided for the convenience of medical providers and UHA members. UHA does not warrant or assure accuracy of such information nor is it intended to be comprehensive in nature. The UHA Formulary is not intended to be a substitute for the knowledge, expertise, skill, and judgment of the medical provider in his/her choice of prescription drugs. UHA assumes no responsibility for the actions or omissions of any medical provider based upon reliance, in whole or in part, on the information contained herein. The medical provider should consult the drug manufacturer's product literature or standard references for more detailed information. The information contained in this document is proprietary. The information may not be copied in whole or in part without the written permission of Umpqua Health Alliance. All rights reserved. This document contains references to brand name prescription drugs that are trademarks or registered trademarks of pharmaceutical manufacturers that are not affiliated with Umpqua Health Alliance. If viewing this formulary via the Internet, please be advised that the formulary is updated periodically and changes may appear prior to their effective date. HOW TO USE THE FORMULARY The medications on the Umpqua Health Alliance (UHA) formulary are grouped into categories depending on the type of medical conditions that they are used to treat. Medications are listed in alphabetical order by the generic name listed in the second column of the drug table. Brand name drugs are capitalized, and generic drugs are listed in lower case.
    [Show full text]
  • Qt3r77r0zg.Pdf
    UCSF UC San Francisco Previously Published Works Title Large-Scale Phenotype-Based Antiepileptic Drug Screening in a Zebrafish Model of Dravet Syndrome Permalink https://escholarship.org/uc/item/3r77r0zg Journal eNeuro, 2(4) ISSN 2373-2822 Authors Dinday, Matthew T Baraban, Scott C Publication Date 2015-07-01 DOI 10.1523/eneuro.0068-15.2015 Peer reviewed eScholarship.org Powered by the California Digital Library University of California New Research Disorders of the Nervous System Large-Scale Phenotype-Based Antiepileptic Drug Screening in a Zebrafish Model of Dravet Syndrome1,2,3 Matthew T. Dinday,1 and Scott C. Baraban1,2 DOI:http://dx.doi.org/10.1523/ENEURO.0068-15.2015 1Department of Neurological Surgery, Epilepsy Research Laboratory, University of California San Francisco, San Francisco, California 94143, 2Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California San Francisco, San Francisco, California 94143 Abstract Mutations in a voltage-gated sodium channel (SCN1A) result in Dravet Syndrome (DS), a catastrophic childhood epilepsy. Zebrafish with a mutation in scn1Lab recapitulate salient phenotypes associated with DS, including seizures, early fatality, and resistance to antiepileptic drugs. To discover new drug candidates for the treatment of DS, we screened a chemical library of ϳ1000 compounds and identified 4 compounds that rescued the behavioral seizure component, including 1 compound (dimethadione) that suppressed associated electrographic seizure activity. Fenfluramine, but not huperzine A, also showed antiepileptic activity in our zebrafish assays. The effectiveness of compounds that block neuronal calcium current (dimethadione) or enhance serotonin signaling (fenfluramine) in our zebrafish model suggests that these may be important therapeutic targets in patients with DS.
    [Show full text]
  • Orthogonal Linear Separation Analysis: an Approach to Decompose the Complex Effects of a Perturbagen
    bioRxiv preprint doi: https://doi.org/10.1101/384446; this version posted August 3, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Title Orthogonal linear separation analysis: an approach to decompose the complex effects of a perturbagen Tadahaya Mizuno ([email protected])*, 1, Setsuo Kinoshita1, 2, Shotaro Maedera1, Takuya Ito1, and Hiroyuki Kusuhara ([email protected])**, 1 1Graduate School of Pharmaceutical Sciences, the University of Tokyo, Bunkyo-ku, Tokyo, 113-0033, Japan 2ProMedico Co., Ltd., Ota-ku, Tokyo, 143-0023, Japan * Corresponding author: Tel: +81-3-5841-4771; E-mail: [email protected] ** Corresponding author: Tel: +81-3-5841-4770; E-mail: [email protected] Running Title: Orthogonal Linear Separation Analysis of Drug Effects Final Character Count: 70,336 characters Abbreviations: AhR, aryl hydrocarbon receptor; BEN, benzamil; BC, benzethonium chloride; BMDM, bone marrow derived macrophages; BSA, bovine serum albumin; CMap, connectivity map; DMEM, Dulbecco’s Modified Eagle’s Medium; FA, factor analysis; FBS, fetal bovine serum; GO, gene ontology; IQ, 2-amino-3-methylimidazo[4,5-f]quinolone; LINCS, library of integrated network-based cellular signatures; LPS, lipopolysaccharide; MET, metiothepin; MEX, metixene; MOA, mode of action; mTORC, mammalian target of rapamycin complex; OLSA, orthogonal linear separation analysis; PCA, principal component analysis; PP, phenazopyridine; SEGR, significant enrichment of GO ratio; TCDD, 2,3,7,8-tetrachlorodibenzo-p-dioxin; WB, western blotting 1 bioRxiv preprint doi: https://doi.org/10.1101/384446; this version posted August 3, 2018.
    [Show full text]
  • Novel Modulation of Adenylyl Cyclase Type 2 Jason Michael Conley Purdue University
    Purdue University Purdue e-Pubs Open Access Dissertations Theses and Dissertations Fall 2013 Novel Modulation of Adenylyl Cyclase Type 2 Jason Michael Conley Purdue University Follow this and additional works at: https://docs.lib.purdue.edu/open_access_dissertations Part of the Medicinal-Pharmaceutical Chemistry Commons Recommended Citation Conley, Jason Michael, "Novel Modulation of Adenylyl Cyclase Type 2" (2013). Open Access Dissertations. 211. https://docs.lib.purdue.edu/open_access_dissertations/211 This document has been made available through Purdue e-Pubs, a service of the Purdue University Libraries. Please contact [email protected] for additional information. Graduate School ETD Form 9 (Revised 12/07) PURDUE UNIVERSITY GRADUATE SCHOOL Thesis/Dissertation Acceptance This is to certify that the thesis/dissertation prepared By Jason Michael Conley Entitled NOVEL MODULATION OF ADENYLYL CYCLASE TYPE 2 Doctor of Philosophy For the degree of Is approved by the final examining committee: Val Watts Chair Gregory Hockerman Ryan Drenan Donald Ready To the best of my knowledge and as understood by the student in the Research Integrity and Copyright Disclaimer (Graduate School Form 20), this thesis/dissertation adheres to the provisions of Purdue University’s “Policy on Integrity in Research” and the use of copyrighted material. Approved by Major Professor(s): ____________________________________Val Watts ____________________________________ Approved by: Jean-Christophe Rochet 08/16/2013 Head of the Graduate Program Date i NOVEL MODULATION OF ADENYLYL CYCLASE TYPE 2 A Dissertation Submitted to the Faculty of Purdue University by Jason Michael Conley In Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy December 2013 Purdue University West Lafayette, Indiana ii For my parents iii ACKNOWLEDGEMENTS I am very grateful for the mentorship of Dr.
    [Show full text]
  • 1 INTRODUCTION the Information Contained in the Umpqua Health Alliance (UHA) Formulary and Its Appendices Is Provided for the Co
    INTRODUCTION The information contained in the Umpqua Health Alliance (UHA) Formulary and its appendices is provided for the convenience of medical providers and UHA members. UHA does not warrant or assure accuracy of such information nor is it intended to be comprehensive in nature. The UHA Formulary is not intended to be a substitute for the knowledge, expertise, skill, and judgment of the medical provider in his/her choice of prescription drugs. UHA assumes no responsibility for the actions or omissions of any medical provider based upon reliance, in whole or in part, on the information contained herein. The medical provider should consult the drug manufacturer's product literature or standard references for more detailed information. The information contained in this document is proprietary. The information may not be copied in whole or in part without the written permission of Umpqua Health Alliance. All rights reserved. This document contains references to brand name prescription drugs that are trademarks or registered trademarks of pharmaceutical manufacturers that are not affiliated with Umpqua Health Alliance. If viewing this formulary via the Internet, please be advised that the formulary is updated periodically and changes may appear prior to their effective date. HOW TO USE THE FORMULARY The medications on the Umpqua Health Alliance (UHA) formulary are grouped into categories depending on the type of medical conditions that they are used to treat. Medications are listed in alphabetical order by the generic name listed in the second column of the drug table. Brand name drugs are capitalized, and generic drugs are listed in lower case.
    [Show full text]
  • Suppl Table 2.Xlsx
    Panic, G. et al. PLOS Neglected Tropical Diseases 9, PRESENT REPORT e0003962 (2015). % Degeneracy Cpds shared with 1,323 drug set d (SRF) after 72 h Categorization Active Inactive M after 72 h current report Amoxapine 4.54 100 Acriflavinium Hydrochloride 3.61 100 1 1 Amodiaquine Dihydrochloride 3.15 81.82 Aprepitant 2.60 100 Atorvastatin Calcium 3.13 76.92 1 1 Benzalkonium Chloride 3.27 100 1 1 Benzyl Benzoate 3.34 83.33 1 1 Beta-naphthol 3.05 100 Bromhexine 2.58 100 Bromocriptine Mesylate 3.23 61.9 Butylated Hydroxyanisole 3.07 100 Chlormidazole 2.88 62.5 chloroxine 2.50 100 1 1 Clioquinol 3.38 100 Cloxyquin 2.54 100 Compactin, Mevastatin 3.51 100 Cyclandelate 2.47 100 Danazol 2.96 58.82 Desipramine 2.46 100 Dichlorophene, Dichlorophen 2.66 100 1 1 Dicyclomine 3.11 100 Digoxin 2.55 100 Disulfiram 2.75 100 1 1 Docetaxel 2.76 92.31 Ebselen 2.78 100 Erythromycin Propionate 4.08 100 Exemestane 2.94 100 Fenbendazole 2.86 100 Fenofibrate 2.78 55.56 Formoterol 2.53 100 hexetidine 3.00 100 1 1 Isobutamben 4.67 100 Ivermectin 3.10 100 d M ≥ 2.47 and Lisuride 2.51 72.73 % Mebhydrolin Naphthalenesulfonate 3.59 72 degeneracy >50 Medroxyprogesterone 2.91 100 Mitotane 3.10 100 Mitoxantrone 4.67 75 Naftopidil Dihydrochloride, Naftopidil 7.29 70.83 nifuroxazide 2.63 100 1 1 Ouabain 3.14 71.43 perhexiline 2.72 100 1 1 Pitavastatin Calcium 4.97 78.26 primaquine 3.25 91.67 1 1 Proflavine Hemisulfate 2.47 100 Promazine 2.52 100 1 1 Propofol 3.08 100 Pyrvinium Pamoate 2.67 100 1 1 Rapamycin 3.64 68.18 reserpine 2.64 60 1 1 Rimantadine 2.51 100 Salinomycin,
    [Show full text]
  • ) (51) International Patent Classification: TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, I
    ) ( (51) International Patent Classification: TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, A61K 31/095 (2006.01) C07D 207/00 (2006.01) EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, A61K 31/10 (2006.01) C07D 207/02 (2006.01) MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, A61K 31/40 (2006.01) C07D 291/04 (2006.01) TR), OAPI (BF, BJ, CF, CG, Cl, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). (21) International Application Number: PCT/US20 18/062287 Published: (22) International Filing Date: — with international search report (Art. 21(3)) 2 1 November 2018 (21. 11.2018) — before the expiration of the time limit for amending the claims and to be republished in the event of receipt of (25) Filing Language: English amendments (Rule 48.2(h)) (26) Publication Language: English — with sequence listing part of description (Rule 5.2(a)) (30) Priority Data: 15/820,334 2 1 November 2017 (21. 11.2017) US 15/820,324 2 1 November 2017 (21. 11.2017) US 16/008,63 1 14 June 2018 (14.06.2018) US 16/008,526 14 June 2018 (14.06.2018) US (71) Applicant: REGEN BIOPHARMA INC. [US/US]; 4700 Spring Street, La Mesa, CA 91942 (US). (72) Inventors: LANDER, Harry, M.; 4700 Spring Street, La Mesa, CA 91942 (US). KOOS, David, R.; 4700 Spring Street, La Mesa, CA 91942 (US). (74) Agent: BRAGINSKY, Philip, Y. et al.; Tarter Krinsky & Drogin LLP, 1350 Broadway, 12th Floor, New York, NY 10018 (US).
    [Show full text]
  • The University of Chicago Microbially-Regulated
    THE UNIVERSITY OF CHICAGO MICROBIALLY-REGULATED INTESTINAL EPITHELIAL HMGB1: STRESS, CELLULAR ENERGY PRODUCTION AND SURVIVAL A DISSERTATION SUBMITTED TO THE FACULTY OF THE DIVISION OF THE BIOLOGICAL SCIENCES AND THE PRITZKER SCHOOL OF MEDICINE IN CANDIDACY FOR THE DEGREE OF DOCTOR OF PHILOSOPHY COMMITTEE ON MOLECULAR METABOLISM AND NUTRITION BY NOELLE PATNO CHICAGO, ILLINOIS JUNE 2017 Copyright © 2017 by Noelle Patno All rights reserved DEDICATION I dedicate this to Jeannette Messer, Ph.D., and Candace Cham, Ph.D., without whose tireless effort and constant attention to me and my work, this thesis would not have been completed. Table of Contents List of Abbreviations ........................................................................................................ v List of Figures ..................................................................................................................ix List of Tables ................................................................................................................... x List of Appendices ...........................................................................................................xi Acknowledgements ........................................................................................................ xii Abstract ......................................................................................................................... xiv Chapter 1: Introduction ...................................................................................................
    [Show full text]
  • Determining the Function of Plasmodium Hemolysin Iii and Discovery of Novel Antimalarial Drugs
    DETERMINING THE FUNCTION OF PLASMODIUM HEMOLYSIN III AND DISCOVERY OF NOVEL ANTIMALARIAL DRUGS by Natalie Robinett A dissertation submitted to Johns Hopkins University in conformity with the requirements for the degree of Doctor of Philosophy Baltimore, Maryland September, 2015 © 2015 Natalie Robinett All Rights Reserved ABSTRACT Elimination of the malaria parasite from endemic areas requires a multi-faceted approach, including development of novel antimalarial drugs and a deeper understanding of parasite-host interactions. Here we describe functional characterization of a Plasmodium hemolysin III (PfHlyIII) along with various approaches to determine whether hemolysin is a virulence factor in malaria, contributing to severe malaria anemia. In addition we also describe two antimalarial drug discovery projects including characterization of novel quinine and quinidine derivatives as efficacious, non-toxic antimalarials, as well as the development of a robust high throughput assay to screen for gametocytocidal compounds. Regarding characterization of Plasmodium hemolysin III, we have evidence for heterologous pore formation of recombinant PfHlyIII in Xenopus and also show expression of soluble native PfHlyIII in asexual blood stage parasites. Together these data support our hypothesis that PfHlyIII may be available upon schizont egress as a cytolytic protein that could damage and increase clearance of bystander erythrocytes. Unexpectedly, genetic disruption of P. berghei HlyIII (PbHlyIII KO) resulted in greater virulence in Balb/c mice leading to an early death phenotype and altered parasitophorous vacuole morphology in the asexual blood stages. We hypothesize that early death in mice infected with the PbHlyIII KO parasite may be a result of altered deformability of infected erythrocytes and increased sequestration leading to brainstem hemorrhage.
    [Show full text]
  • Electronic Supplementary Material (ESI) for Organic & Biomolecular
    Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry.
    [Show full text]