Photograph by author, Gina M.A. Carroll Metabolic Stable Isotope Fractionation: Biogeochemical Approaches to Diagnosing Sickle Cell and Thalassemia Anemia in the Archaeological Record MSc Thesis Faculty of Archaeology MSc Proefschrift Faculteit der Archaeologie Gina M.A. Carroll 1 Photograph by Gina. M.A. Carroll Taken with permission from the Municipal Museum of Écija, Spain April 2014 Gina M.A. Carroll Alberta, Canada Leiden, The Netherlands [email protected] 1 Metabolic Stable Isotope Fractionation: Biogeochemical Approaches to Diagnosing Sickle Cell and Thalassemia Anemia in the Archaeological Record. MSc Thesis MSc Proefschrift Gina M.A. Carroll Human Osteology and Funerary s1371266 Archaeology MSc Thesis Archaeology University of Leiden Faculty of Archaeology ARCH 1044WY Prof. Dr. Waters-Rist Leiden, The Netherlands & Prof. Dr. Inskip Leiden, 26 May 2015 Final Draft. 2 TABLE OF CONTENTS DEDICATIONS ...................................................................................................................... 9 ACKNOWLEDGEMENTS .................................................................................................. 10 CHAPTER 1 INTRODUCTION ....................................................................... 12-30 1. BRIEF HISTORY OF ARCHAEOLOGICAL RESEARCH ........................................ 13 1.1. The Anemias in Archaeology ....................................................... 14 1.2. The Application of Stable Isotopes in Palaeopathology ............... 18 2. HYPOTHESIS ................................................................................................. 19 3. DEFINITIONS AND BOUNDARIES .................................................................. 21 4. BIOMEDICAL APPROACHES TO ARCHAEOLOGY: SICKLE CELL AND THALASSEMIA .............................................................................................. 24 4.1. The Disease Model ....................................................................... 24 4.2. The Anemia Model ....................................................................... 25 4.3. Sample Selection .......................................................................... 26 5. RESEARCH GOALS ........................................................................................ 26 6. CHAPTER OUTLINE ....................................................................................... 27 6.1. Chapter 2: Introduction to Stable Oxygen, Carbon and Nitrogen Isotopes ......................................................................................... 27 6.2. Chapter 3: Introduction to the Genetic and Pathophysiological Mechanisms of Sickle Cell and Thalassemia ............................... 27 6.3. Chapter 4: Physiological Mechanisms of Stable Isotope Diffusion, Fixation and Fractionation during Total Respiration .................... 28 6.4. Chapter 5: Literature Review: Metabolic Fractionation of Stable Oxygen and Carbon Isotopes in Chronic Anemias ....................... 28 6.5. Chapter 6: Pathophysiological Fractionation of Stable Isotopes Associated with Rates of Protein Turnover and Energy Expenditure ................................................................................... 28 6.6. Chapter 7: The Socio-Religious History of Écija, Spain: Islam and Isotopes ......................................................................................... 29 6.7. Chapter 8: Methods and Materials ................................................ 29 6.8. Chapter 9: Results ......................................................................... 29 6.9. Chapter 10: Discussion ................................................................. 30 6.10 Chapter 11: Summary and Conclusion. ...................................... 30 CHAPTER 2 INTRODUCTION TO STABLE OXYGEN, CARBON AND NITROGEN ISOTOPES ................................................................................................ 31-43 1. INTRODUCTION TO ATOMIC THEORY: RADIOACTIVE AND STABLE ISOTOPES ......... 31 1.1. Chemical and Molecular Properties of Stable Isotopes ............................ 33 1.2. Primary and Secondary Stable Isotopes .................................................... 33 2. STABLE ISOTOPE FRACTIONATION .......................................................................... 34 2.1. Isotopic Fractionation and the Archaeological Record ............................. 36 2.2. The Isotopic Composition of Tissues: Understanding Delta (δ) Values ... 38 3 2.3. Standards ................................................................................................... 38 3. STABLE ISOTOPE ASSIMILATION INTO BIOLOGICAL TISSUES: THE OSTEOLOGICAL RESERVOIR .............................................................................................................. 38 3.1. The Osteological Reservoirs: Hydroxyapatite and Bone Collagen ........... 39 3.2. Stable Isotope Analyses and Anemia ........................................................ 41 4. SUMMARY AND CONCLUSIONS ............................................................................... 42 4.1. Notes ......................................................................................................... 42 CHAPTER 3 INTRODUCTION TO THE GENETIC AND PATHOPHYSIOLOGICAL MECHANISMS OF SICKLE CELL AND THALASSEMIA ............................................................................................................. 44-67 1. HEMOGLOBIN .......................................................................................................... 44 1.1. Globin Chains ............................................................................................ 45 1.2. HbF and HbA2 Hemoglobin: ɣ and δ Polypeptides ................................. 45 2. Thalassemia ............................................................................................................ 46 2.1. Alpha Thalassemias................................................................................... 47 2.2. Beta Thalassemias ..................................................................................... 51 3. SICKLE CELL........................................................................................................... 54 3.1. Homozygotic Sickle Cell .......................................................................... 55 3.2. Heterozygotic Sickle Cell .......................................................................... 57 4. MODIFYING INFLUENCES: HBF PERSISTENCE, HBA2, AND SICKLING-THALASSEMIAS ................................................................................................................................ 58 5. SICKLE CELL AND THALASSEMIA IN ÉCIJA ............................................................. 59 5.1. Skeletal Signatures .................................................................................... 59 5.2. Contemporary Genetic Distribution .......................................................... 61 5.3. Malaria and Genetic Anemias ................................................................... 63 6. SUMMARY AND CONCLUSIONS ............................................................................... 65 CHAPTER 4 PHYSIOLOGICAL MECHANISMS OF STABLE ISOTOPE DIFFUSION, FIXATION AND FRACTIONATION DURING TOTAL RESPIRATION ............................................................................................................................................ 68-91 1. DEFINITIONS AND CONCEPTS .................................................................................. 69 2. PHYSIOLOGICAL MECHANISMS OF STABLE OXYGEN, NITROGEN AND CARBON ISOTOPE DIFFUSION, FIXATION AND METABOLIZATION DURING TOTAL RESPIRATION ........................................................................................................... 70 2.1. Total Respiration and Stable Oxygen Fractionation ................................. 70 2.2. Cellular Respiration and Stable Carbon Fractionation .............................. 72 2.3. Total Respiration, Inflammation and Stable Nitrogen Fractionation ........ 74 3. CONTRIBUTION OF FRACTIONATION FACTORS TO ISOTOPIC SIGNATURES .............. 75 4. HEMOGLOBIN: THE FIXATION OF OXYGEN, NITROGEN AND CARBON MOLECULES IN SICKLE CELL AND THALASSEMIA ANEMIA .............................................................. 77 4.1. Hemolysis: Effects on Blood Oxygen, Carbon and Nitrogen Concentrations ........................................................................................... 77 4.2. Inflammation ............................................................................................. 82 4.3. Summary ................................................................................................... 82 4 5. TOTAL RESPIRATION: CARDIORESPIRATORY COMPLICATIONS AND RATES OF OXYGEN AND CARBON DIFFUSION AND FIXATION .................................................. 83 6. DISCUSSION ............................................................................................................. 86 6.1. Stable Isotope Fractionation in Anemics: Hypothesis .............................. 86 6.2. Anemia Related Stable Isotope Fractionation in Archaeological Populations ................................................................................................ 88 7. SUMMARY AND CONCLUSIONS ............................................................................... 89 7.1. Summary ................................................................................................... 89 7.2. Conclusions ..............................................................................................