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Evaluation of New Techniques for Zinc and Cortisol

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Evaluation of New Techniques for Zinc and Cortisol EVALUATION OF NEW TECHNIQUES FOR ZINC AND CORTISOL ASSESSMENT WITH A PLACEBO-CONTROLLED ZINC SUPPLEMENTATION TRIAL IN A SUBSAMPLE OF ETHIOPIAN WOMEN By MAYA LUCIA JORAY Bachelor of Science in Food Technology University of Applied Sciences Wädenswil, Zürich, Switzerland 2001 Master of Science in International Studies Oklahoma State University Stillwater, OK 2004 Submitted to the Faculty of the Graduate College of the Oklahoma State University in partial fulfillment of the requirements for the Degree of DOCTOR OF PHILOSOPHY July, 2011 EVALUATION OF NEW TECHNIQUES FOR ZINC AND CORTISOL ASSESSMENT WITH A PLACEBO-CONTROLLED ZINC SUPPLEMENTATION TRIAL IN A SUBSAMPLE OF ETHIOPIAN WOMEN Dissertation Approved: Dr. B. J. Stoecker Dissertation Adviser Dr. S. L. Clarke Dr. E. Lucas Dr. P. Rayas Duarte Dr. M. E. Payton Interim Dean of the Graduate College ii TABLE OF CONTENTS Chapter Page I. INTRODUCTION ......................................................................................................1 Purpose ....................................................................................................................4 Objective .................................................................................................................5 Null hypotheses ........................................................................................................5 Assumptions and limitations ....................................................................................6 Organization of the dissertation ...............................................................................7 II. REVIEW OF LITERATURE....................................................................................8 Study area and population ........................................................................................8 Study population ..............................................................................................10 Nutritional status and diet of the study population ..........................................11 Zinc .......................................................................................................................17 Physiological function of zinc .........................................................................17 Endogenous and exogenous sources of zinc ....................................................21 Dietary factors leading to zinc deficiency .......................................................22 Requirements of zinc .......................................................................................24 Impact of zinc deficiency and its indicators ....................................................26 Zinc deficiency in the study population ...........................................................30 Supplementation with zinc and possible outcomes .........................................31 Zinc digestion, absorption and metabolism .....................................................32 Transport of zinc within the cells.....................................................................35 Transcription factors involved in zinc homeostasis .........................................36 Krüppel-like factor 4 ........................................................................................38 Specific role of Zip4 ........................................................................................40 Other zinc transporters .....................................................................................42 Storage and excretion of zinc ...........................................................................44 Assessment of zinc status.................................................................................47 Novel assessment method of zinc ....................................................................50 Cortisol ...................................................................................................................52 Physiological function of cortisol ....................................................................52 Physiological cortisol levels ............................................................................55 Assessment methods of cortisol .......................................................................55 Interaction of zinc and cortisol ........................................................................58 iii III. MATERIALS AND METHODS ...........................................................................61 Location and study design .....................................................................................61 Study area and population ................................................................................61 Ethical approval ...............................................................................................62 Study design .....................................................................................................63 Pre-survey preparations ...................................................................................63 Zinc supplements and placebo .........................................................................64 Sampling and data collection ...........................................................................64 Biochemical analysis ............................................................................................66 Zinc in plasma ..................................................................................................66 Zinc in erythrocyte ...........................................................................................66 Zip4 transporter and metallothionein in buccal cells .......................................67 Cortisol in saliva, serum, and urine .................................................................69 Iodine in urine ..................................................................................................71 Ferritin in serum ..............................................................................................72 Comet assay in whole blood ............................................................................72 α-1-acid glycoprotein in serum ........................................................................73 High sensitivity C-reactive protein in serum ...................................................74 Ferric reducing antioxidative power measured in serum .................................75 Albumin in serum ............................................................................................76 Total protein inurine ........................................................................................77 Statistical analysis .................................................................................................78 IV. ASSESSMENT OF CHANGES IN mRNA EXPRESSION IN BUCCAL CELLS AT BASELINE AND AFTER A PLACEBO-CONTROLLED ZINC SUPPLEMENTATION TRIAL IN SIDAMA, ETHIOPIA ..................................79 Abstract ..................................................................................................................79 Introduction ............................................................................................................80 Volunteers, material and methods..........................................................................83 Volunteers Stillwater, OK ................................................................................83 Volunteers Finichawa, Sidama, Ethiopia .........................................................83 Study design, supplementation, sample collection, (Finichawa, Sidama, Ethiopia) ..........................................................................................................83 Buccal cell sampling .......................................................................................84 Assessment of plasma zinc .............................................................................85 RNA extraction from RNAlater treated samples ............................................85 Ethical approval ..............................................................................................86 Results ....................................................................................................................86 Discussion ..............................................................................................................90 References ............................................................................................................105 iv V. CORTISOL LEVELS MEASURED IN DIFFERENT COMPARTMENTS IN WOMEN FROM SIDAMA, ETHIOPIA ............................................................107 Abstract ...............................................................................................................107 Introduction .........................................................................................................108 Volunteers, material and methods .......................................................................112 Volunteers, study design, supplementation, sample collection ....................112 Cortisol in urine, serum and saliva ...............................................................112 Iodine in urine ...............................................................................................113 High sensitivity C-reactive protein in serum ................................................114 α-1-acid glycoprotein ....................................................................................114 Albumin in serum .........................................................................................115
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