CORE Metadata, citation and similar papers at core.ac.uk Provided by Federation ResearchOnline 1 NEUROTROPHIC BIOMARKER CHANGE AFTER PHYSICAL ACTIVITY AND MINDFULNESS INTERVENTIONS. A thesis submitted for the degree of Master of Science at Federation University, Ballarat, Australia. Gina England. November 2017. 2 ATTRIBUTION OF COLLABORATIVE CONTRIBUTION. The Wellness project was established by Prof. Britt Klein at Faculty of Health & Centre for Biopsychosocial eHealth Research and Innovation, Federation University (Ballarat) and staff and students, who advertised, recruited and organised attendance on site for assessments and blood collection. The original study question and hypothesis involved in analysis of neurotrophin mRNA change in association with mood was established by the candidate in association with Prof. Fadi Charchar (Faculty of Science and Technology, Federation University, Ballarat) Blood collection was carried out by the candidate on the Ballarat campus along with sample preparation and stabilisation prior to the RNA extraction. Priscilla Prestes and the candidate extracted the RNA and Priscilla subsequently prepared the cDNA and processed the raw data output from the qPCR analysis in the FOST Genetics lab (Ballarat). The candidate completed all gene expression assays, raw data analyses and statistical analyses. Except where explicit reference is made in the text of the thesis, this thesis contains no material published elsewhere, nor work relied upon or used without due acknowledgement in the main text or references listed. This thesis does not include work with copyright provisions or requiring copyright approvals. 3 ACKNOWLEDGEMENTS. I would like to express my deepest thanks to my primary supervisor Prof. Fadi Charchar for facilitating the completion of a journey which began and terminated prematurely many years ago. Without your encouragement and guidance I would not have had the courage for this endeavour. Your logical approach to complexity relieves stress, calms anxiety and reduces symptoms of depression, perhaps we should study the Charchar Intervention. To Prof Britt Klein, thank you for your enthusiasm for any direction I wanted to take and swift and encouraging feedback on all the manuscripts for review. Always positive and helpful critique without the criticism, at all times, even when you are completely inundated. To Priscilla Prestes my lab buddy and teacher of all things PCR related, and frankly anything else I needed to know or had forgotten about returning to lab work. You made it fun, and knowing Priscilla was in the building took the stress levels down. I hope my endeavour might in some way inspire yours. Finally to my wonderful husband Paul who has rather more often than I would like to admit rescued this manuscript from computer malfunctions, formatting aberrations and various careless misdemeanours with humour and good spirit. At all times you were more confident in my ability than I was in myself. Thank you. 4 CONTENTS Attribution of collaborative contribution. .................................................................................. 2 Acknowledgements. ................................................................................................................ 3 Abstract ................................................................................................................................... 6 Introduction .................................................................................................................. 8 Depression ................................................................................................................. 12 Anxiety ....................................................................................................................... 14 Stress ......................................................................................................................... 15 Integrating Inflammation and Behaviour .................................................................... 18 Wellness ..................................................................................................................... 20 Modalities of Symptom Relief ..................................................................................... 22 Mindfulness ......................................................................................................... 22 Physical Exercise ................................................................................................ 28 Significance of the neurotrophins ............................................................................... 32 BDNF .................................................................................................................. 34 NGF .................................................................................................................... 39 FGF2. .................................................................................................................. 44 The use of mRNA as a biomarker .............................................................................. 49 Aim ......................................................................................................................... 51 Hypothesis .............................................................................................................. 52 Materials and Methods ........................................................................................... 52 5 Participant Selection ........................................................................................ 53 Pre-intervention Assessment .......................................................................... 54 Intervention period ........................................................................................... 55 Components of the Interventions .................................................................... 56 DASS Measurements During Intervention. ..................................................... 57 General Laboratory Protocols ......................................................................... 58 Results .................................................................................................................... 67 DASS Analysis ................................................................................................ 70 Neurotrophin Analysis ..................................................................................... 74 Investigations into the Amplification Curve for NGF ........................................ 78 Discussion .............................................................................................................. 80 Depression, Anxiety and Stress association with Neurotrophin expression and Mindfulness .................................................................................................................... 80 Depression Anxiety and Stress association with Neurotrophin expression and Physical Activity ............................................................................................................. 83 Intervention FGF2 and NGF expression. ........................................................ 85 Strengths and limitations ........................................................................................ 86 Strengths ......................................................................................................... 86 Limitations ....................................................................................................... 87 Conclusion .............................................................................................................. 90 References ............................................................................................................. 92 Appendix A The DASS………………………………………………………...…...…………..…107 6 ABSTRACT BACKGROUND AND AIM AND HYPOTHESIS: BDNF, FGF2 and NGF are neurotrophins associated with neuroplasticity, nervous system development and psychiatric disorder in the literature. BDNF in particular is suggested as a useful biomarker of mood disorder. Both mindfulness and physical activity are shown to improve mood, reduce stress and are widely used as part of a multi-component treatment approach, reducing distressing symptoms of mood and affect disorders. The utility of protein level as a biomarker has been controversial in the literature following issues concerning the assessment of peripheral levels as a proxy for central levels. The principal aim of this study was to investigate the gene expression of three neurotrophins BDNF, FGF2 and NGF as potential biomarkers of mood disorder, at an early stage of these disorders, which are now widely recognised as having pathogenesis related to dysregulation in the neuro-immuno-endocrine axis. In addition, the study will explore the effect of both physical activity and mindfulness on neurotrophin expression clarifying the associations between the success of these interventions seen in the literature and their effect on the change of neurotrophin expression. Current literature reports increased levels of BDNF protein both centrally and peripherally following mood disorder treatment and participation in both physical exercise and mindfulness activities. Based on similarity of structure and function amongst the three neurotrophins, this thesis will hypothesize an increase in BDNF and potentially FGF2 and NGF mRNA following participation in the two interventional modules designed to improve wellbeing in clinical and non-clinical