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Mountain Lab: Studying the effects of stress and extreme conditions on human physiology

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The human body is an amazingly complicated machine, capable of adapting and responding to various stressors and environmental conditions. Even in extreme situations the body is able to adjust core physiological processes and systems to ensure optimal function, and ultimately, survival. When studying human physiological response the most basic measurements, such as ECG and respiration, can hold huge amounts of information. But, their value is much greater when integrated with other physiological measurements such as blood pressure, oxygen saturation and respiratory gas concentrations.

However, accurate co-registration of physiological data is no trivial pursuit. Moreover, the complexity of such research endeavors is compounded when we venture out of traditional laboratory spaces and seek to study human response and adaptation in extreme environments. Sensors and systems must offer practical application and reliable data collection -- moreover, data storage and management is of critical importance.

In this webinar sponsored by ADInstruments, Dr. Trevor Day, Associate Professor of Physiology at Mount Royal University in Calgary Alberta, shares his research on the effects of tilt, exercise and high altitude on respiratory sinus arrhythmia (RSA). These case studies serve as representations of more complex applications of human physiologic monitoring, in particular, his trek to Everest Base Camp where he and his research team monitored and tracked acclimatization in the context of high altitude hypoxia. During this expedition multiple physiological measures were recorded simultaneously on both rest and exercise days in order to test for signs of altitude sickness. Dr. Day shares his experiences from this exciting study and others conducted at his lab at Mount Royal to offer perspective regarding the importance of being able to record and integrate multiple data streams simultaneously.

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Mountain Lab: Studying the effects of stress and extreme conditions on human physiology

  1. 1. Mountain Lab: Studying the effects of stress and extreme conditions on human physiology A webinar discussing the effects of tilt, exercise and high altitude on human cardiorespiratory and autonomic nervous systems, as studied in traditional laboratory settings and on location at Everest Base Camp.
  2. 2. InsideScientific is an online educational environment designed for life science researchers. Our goal is to aid in the sharing and distribution of scientific information regarding innovative technologies, protocols, research tools and laboratory services. JOIN FOR FREE AT WWW.INSIDESCIENTIFIC.COM
  3. 3. Mountain Lab: Studying the effects of stress and extreme conditions on human physiology Dr. Trevor Day Associate Professor of Physiology Department of Biology Faculty of Science and Technology Mount Royal University tday@mtroyal.ca
  4. 4. Acknowledgements Trainees: Jeff Baden B.Sc. M.Sc. Maria Abrosimova B.Sc. Gary Saran B.Sc. Lauren Lavoie B.Sc. Jamie Pfoh B.Sc. Christina Bruce B.Sc. Kennedy Borle B.Sc. Andrea Linares Rachelle Brandt B.Sc. Kartika Tjandra Ph.D. Michael Tymko M.Sc. Rachel Skow M.Sc. Lindsey Boulet B.Sc. A special thank you to our research participants, MRU Human Research Ethics Board and Nepal Health Research Council Collaborators: Funding & Support:
  5. 5. Calgary, Alberta, Canada
  6. 6. Mount Royal University
  7. 7. Chemoreflex Control of Breathing • Central respiratory chemoreflex • Peripheral respiratory chemoreflex • Central-peripheral chemoreceptor interaction • Intermittent hypoxia • High altitude hypoxia and acclimatization Cerebral Blood Flow Regulation • Cerebral autoregulation • Cerebrovascular CO2 reactivity • Neurovascular coupling Hypovolemia and Tilt • Cerebrovascular regulation • Baroreflex responses • Respiratory sinus arrhythmia (RSA) MRU INTEGRATIVE Physiology lab Department of Biology, Faculty of Science and Technology
  8. 8. Kidneys Heart Lungs Brain Stress Stress Stress Stress
  9. 9. List of Acronyms • ECG: Electrocardiogram • HR: Heart rate • BPM: beats per minute • RSA: Respiratory sinus arrhythmia • VTI: Inspired tidal volume (L) • FVC: Forced vital capacity (L) • MAP: Mean Arterial Pressure (i.e., blood pressure; mm Hg) • TCD: Transcranial Doppler ultrasound (for brain blood flow) • MCAv: Middle cerebral artery velocity (using ultrasound; cm/s) • Q: Cardiac output (L/min) • V/Q: Ratio relating alveolar ventilation and perfusion of the lung • PETCO2: pressure of end-tidal CO2 (Torr) • HUT and HDT: Head-up and head-down tilt
  10. 10. Novel Integrated Tilt Table-Lower Body Negative Pressure Box (LBNP) • Built by Michael Tymko (M.Sc.; now PhD student UBC) • Superimposes tilt and LBNP stressors • Tilt table allows HUT and HDT • LBNP chamber creates a negative pressure to translocate blood volume toward the lower body
  11. 11. 2014 Alberta Science and Technology (ASTech) Young Innovator Award
  12. 12. Michael Tymko recently published an “instruction manual” on constructing LBNP chambers (Nov 2016, In press).
  13. 13. “The effects of superimposed tilt and lower body negative pressure on anterior and posterior cerebral circulations” Tymko et al., 2016
  14. 14. 2015 American Physiological Society ADInstruments Macknight Early Career Innovative Educator Award APS President David Pollock and Anthony Macknight of ADI present the ADInstruments Macknight Early Career Innovative Educator Award to Trevor A. Day
  15. 15. Respiratory Sinus Arrhythmia (RSA) • RSA is the normal fluctuation of heart rate in phase with the respiratory cycle • Inspiration = increase in HR • Expiration = decrease in HR • HR quantified from the ECG • The “peak-valley” of the HR tracing quantifies RSA magnitude
  16. 16. • These signals are processed in ADI LabChart Pro from analog inputs • IHR from ECG • MAP from a raw finometer input • MCAv mean from TCD • VTI from respiratory flow • PETCO2 from breath by breath expired gas analyzer • Note that MAP and MCAv fluctuate in phase with RSA RSA affects blood pressure and brain blood flow
  17. 17. Possible mechanisms underlying Respiratory sinus arrhythmia (RSA) RSA magnitude is thought to represent the dominance of parasympathetic nervous system tone at rest. Possible mechanisms include: 1. Firing of respiratory neurons impacting the firing of cardiac motor neurons in the brainstem. 2. Stretch receptors in the lungs and chest wall. 3. Changes in blood pressure with breathing acting on arterial baroreceptors (carotid and aortic sinus). 4. Changes in venous return and cardiac loading with breathing stimulating low pressure receptors in the right atrium (Bainbridge reflex).
  18. 18. Respiratory Pump
  19. 19. Inspiration Increased heart rate Increased venous return Increased right atrial pressure Inhibition of medullary cardiac neurons and vagal withdrawal Stimulation of stretch receptors in right atria and pulmonary artery
  20. 20. Expiration Decreased heart rate Decreased venous return Decreased right atrial pressure No inhibition of medullary cardiac neurons and increased vagal tone Less stimulation of stretch receptors in right atria
  21. 21. Factors Modulating RSA Magnitude? Tidal volume Nervous system activation Blood gas levels Fitness level Respiratory frequency Age RSA
  22. 22. Possible Utility of RSA? RSA may increase pulmonary gas exchange efficiency through improved V/Q matching Ventilation Perfusion Inspiration HR and Q increase Expiration HR and Q decrease
  23. 23. Tilt Exercise Hypoxia
  24. 24. Tilt and blood volume distribution • Tilt causes gravity-dependent redistribution of blood volume • Standing or HUT translocates up to 1L of blood volume toward the lower extremities • HDT translocates blood into the central cavity, increasing venous return and cardiac loading Trendelenburg position
  25. 25. • Gelinas et al., 2012 Aviat Space Environ Med • Skow et al., 2013 Resp Physiol Neurobiol • Skow et al., 2014 Prog Brain Res • Tymko et al., 2015 Exp Physiol We investigated the effects of steady-state tilt on respiratory and cerebrovascular regulation. Previous tilt studies in the lab
  26. 26. Baden et al., 2014 Aviat Space Environ Med Case Report: 45 Degree Head Down Tilt
  27. 27. Case Report: 45 Degree Head Down Tilt • Sinus arrhythmia • Note the P waves (red arrows) • NOT pathological
  28. 28. Badenetal.,2014AviatSpaceEnvironMed
  29. 29. Experiment #1: Tilt and RSA Aim: To explore the relationship between superimposed gravity- dependent and inspiration-dependent cardiac filling on RSA magnitude. Hypothesis: Superimposed gravity- and inspiration-dependent cardiac loading will increase RSA magnitude in a synergistic fashion.
  30. 30. Methods 10%, 20%, 30%, 40%, and 50% of FVC RANDOMIZED RANDOMIZED 40o HDT 40o HUT FVC (x3) n=19
  31. 31. Analysis • Peak-valley method • Data from 5 of the most accurate, consecutive breaths • Correlation between VTI and RSA magnitude • RSA magnitude plotted against each targeted VTI (% FVC) • Linear regression of RSA magnitude against VTI • Slopes calculated to quantify “RSA reactivity” 20% FVC 40% FVC Abrosimova et al., Manuscript in Preparation
  32. 32. HUT; r = 0.64; P<0.001 HDT; r = 0.53; P<0.001 RSA magnitude is correlated with VTI Abrosimova et al., Manuscript in Preparation
  33. 33. HUT=0.43 HDT=0.33R2=0.99 R2=0.99 P=0.02 “RSA reactivity” in response to increases in VTI is linear Abrosimova et al., Manuscript in Preparation Response slopes are tilt-dependent
  34. 34. Summary • RSA magnitude increases linearly with increases in VTI (“RSA reactivity”) • RSA reactivity is not increased with HDT • RSA reactivity is decreased in HDT, likely do to sympathetic NS modulation • Question: Can we test RSA reactivity during another stressor where venous return is increased and the sympathetic NS is activated?
  35. 35. Skeletal Muscle Pump
  36. 36. Aim: To explore the relationship between superimposed exercise stress (with skeletal muscle pump activity and sympathetic nervous system activation) and inspiration-dependent cardiac filling on RSA magnitude. Hypothesis: Sympathetic activation during exercise will reduce RSA magnitude, despite superimposed inspiratory-dependent and skeletal muscle pump cardiac filling. Experiment #2: Exercise and RSA
  37. 37. Methods and Instrumentation • Participants (n=13) instrumented for respiratory volumes and heart rate • Seated on a cycle ergomenter • Participant feedback on respiratory volume via computer screen • RSA trials repeated at rest and during exercise
  38. 38. Protocol Lavoie et al., Manuscript in Preparation
  39. 39. Results – Raw Traces Rest Exercise Lavoie et al., Manuscript in Preparation
  40. 40. ResultsResults – RSA reactivity is reduced during exercise Rest Exercise Lavoie et al., Manuscript in Preparation
  41. 41. Results P = 0.001 • RSA reactivity is eliminated during exercise • This is despite an increase in venous return during exercise • RSA is likely NOT driven by increases in venous return. Lavoie et al., Manuscript in Preparation
  42. 42. • RSA is maintained during exercise. • However, RSA reactivity is eliminated during exercise, despite increases in venous return, likely because of increased sympathetic activity. • Questions: Will RSA be affected by acclimatization to high altitude hypoxia? Could RSA reactivity magnitude affect V/Q matching and oxygenation during hypoxic stress? Summary
  43. 43. Integrate and analyze all your data streams in one place Setting the pace for Exercise Research • Wireless physiological monitoring and EMG • Metabolic Systems • Accelerometry • Goniometers • Human NIBP • Stimulators
  44. 44. Himalayan Mountain Range - Tibet/Nepal Mount Everest 8848 m (29,028 ft) Atmospheric Pressure = 253 mm Hg Available Oxygen ~33% of Sea Level
  45. 45. 0 100 200 300 400 500 600 700 800 0 1 2 3 4 5 6 7 8 9 10 GasPressure(mmHg) Altitude (kilometres) Patm (mmHg) PO2 (mmHg) The Relationship Between Altitude and Relative Gas Pressures Day TA (2010). Human Adaptation to High Altitude Hypoxia: Getting High. Biology on the Cutting Edge: Canadian Research and Issues around the Globe. (pp. 117-122) Pearson Education Canada, Toronto, Ontario. Vancouver Calgary Airplane Cabin Everest Half the available oxygen of sea level
  46. 46. Aim: To explore the relationship between superimposed high altitude hypoxia and inspiration-dependent cardiac filling on RSA. Hypothesis #1: Increases in sympathetic nervous system during high altitude hypoxia will reduce RSA magnitude (similar to exercise). Hypothesis #2: Larger RSA magnitude will improve oxygenation through improved V/Q matching at altitude. Experiment #3: High Altitude Hypoxia and RSA
  47. 47. Everest Base Camp (EBC) Trek
  48. 48. Why Nepal?
  49. 49. Altitude Comparisons: Banff and Mt. Rundle = Kathmandu and Lukla
  50. 50. May 2016 23 participants recruited including nine paid trainees from MRU, collaborators, industry partners (ADI) and community members from across Canada, USA, New Zealand and Ireland. Ethical Clearance: • Mount Royal University Human Research Ethics Board 2015-26b • Nepal Health Research Council 96/2016 Objective: • A fast and light approach to high altitude acclimatization on a trek to Everest Base Camp
  51. 51. Pelican cases packed outside the Lab April 29, 2016
  52. 52. Calgary Airport April 30, 2016
  53. 53. Kathmandu (1400m) Monjo (2835m) Namche (3440m) Tengboche (3860m) Pheriche (4370m) Lobuche (4940m) Gorak Shep (5160m) Pheriche (4370m) Pangboche (3985m) Kunde (3840m) Namche (3440m) Phakding (2610m) Lukla (2860m) Kathmandu (1400m) 0 500 1,000 1,500 2,000 2,500 3,000 3,500 4,000 4,500 5,000 5,500 6,000 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 Day Acetazolamide (Diamox) 125mg PO BID Altitude(m) Ascent-DescentProfile–Nepal2016
  54. 54. Lukla Airport (2800m)
  55. 55. Daily Measures: Nepal May 2016 • Between 6-8 am, before breakfast, following one night at that altitude. • Heart rate and peripheral oxygen saturation • Respiratory rate and pressure of end-tidal CO2 • Blood pressure • Acute Mountain Sickness scores (Lake Louise Scoring system) • Actigraph accelerometers for daily activity and sleep disturbances • Collected on ascent and descent • n=21
  56. 56. Rest Day Measures: Nepal May 2016 • On rest days during ascent between 10 AM and 5 PM • Calgary (1045m), Namche (3440m), Tengboche (3860m) and Pheriche (4370m) • [Hemoglobin] and hematocrit, urine pH and renal reactivity, voluntary breath holding, ventilatory acclimatization, heart rate variability (RSA reactivity) • n=12
  57. 57. Namche (3440m)
  58. 58. Namche (3440m)
  59. 59. Tengboche (3860m)
  60. 60. Pheriche (4370m)
  61. 61. Pheriche (4370m)
  62. 62. Poincare Plot – Quantification of heart rate variability Saran et al., Manuscript in Preparation
  63. 63. PoincarePlotsandAltitude Saran et al., Manuscript in Preparation
  64. 64. RSA during spontaneous breathing via SD1/SD2 ratio Resting RSA magnitude is not changed with high altitude ascent Saran et al., Manuscript in Preparation
  65. 65. • RSA quantified using the peak- valley approach • Participant targets inspired volume through computer screen feedback RSA and targeted VTI
  66. 66. RSA Reactivity Slopes and Altitude • RSA protocol during ascent • We plotted RSA magnitude against %FVC • Slopes quantify “RSA Reactivity” • Slopes appear unchanged with altitude Saran et al., Manuscript in Preparation
  67. 67. Results - RSA reactivity magnitude is not altitude dependent Saran et al., Manuscript in Preparation
  68. 68. Gorak Shep (5160m)
  69. 69. 70 75 80 85 90 95 100 Kathmandu (1400m) Monjo (2840m) Namche (3440m) Tengboche (3860m) Pheriche (4370m) Lobuche (4940m) Gorak Shep (5160m) PeripheralOxygenSaturation(%) Location and Altitude Oxygen Saturation and Altitude • SpO2 (%) measured every morning during ascent • Note the reduction in SpO2 (%) with increases in altitude
  70. 70. RSA and V/Q matching hypothesis
  71. 71. The effects of RSA magnitude on oxygen saturation
  72. 72. • Resting RSA magnitude is unchanged with acclimatization to high altitude (Poincare plots) • RSA reactivity to targeted increases in VTI is also unchanged with acclimatization to high altitude • RSA magnitude does not improve oxygen saturation in the context of hypoxia, suggesting V/Q matching hypothesis is incorrect. Summary
  73. 73. Summit of Kala Patthar (~5600m)
  74. 74. Acute Mountain Sickness (AMS)
  75. 75. Lukla (2800m)
  76. 76. Lukla Airport (2800m)
  77. 77. Kathmandu (1400m)
  78. 78. Calgary
  79. 79. Research in Austere Environments… a balance between FEASABILITY and NOVELTY
  80. 80. Research in Austere Environments • Building the right team • Organization and safety • Managing expectations: the needs of the individual/team with needs of the researchers
  81. 81. • Cultural sensitivity • Personal and interpersonal perspectives • Staying positive and optimistic • Keeping your sense of humour Research in Austere Environments
  82. 82. • Creativity • Improvisation • Serendipity • Persistence • Compromise • Problem solving • Responsive to new opportunities • Expect the unexpected • Know the limitations of your gear • Power? Research in Austere Environments
  83. 83. Undergradute Students!
  84. 84. Lake Louise AMS Scoring System [Roach et al., 1993] [2] Moderate Headache [3] Severe Headache. Incapacitating Score = 2. Gastrointestinal Symptoms [0] Good Appetite [1] Poor Appetite/Nausea [2] Moderate Nausea/Vomiting [3] Severe. Incapacitating Nausea and Vomiting Score = 3. Fatigue and/or weakness [0] Not Tired or Weak [1] Mild Fatigue/Weakness [2] Moderate Fatigue/Weakness [3] Severe Fatigue/Weakness Score = 4. Dizziness/light-headedness [0] None [1] Mild [2] Moderate [3] Severe. Incapacitating Score = 5. Difficulty sleeping [0] Slept as well as usual [1] Did not sleep as well as usual [2] Woke many times. Poor night’s sleep [3] Could not sleep at all Score = Sum 1-5 Total AMS Score = Sum 1-4 Total AMS Score = LAKE LOUISE AMS SCORING SYSTEM Name: Date: Location and Altitude: Instructions: Please circle the number of each item to correspond to HOW YOU FEEL AT THIS PRESENT MOMENT. PLEASE ANSWER EVERY ITEM. If you do not have the specific symptom, please circle [0]. Self-Assessment Score 1. Headache [0] None at all [1] Mild Headache [2] Moderate Headache [3] Severe Headache. Incapacitating Score = 2. Gastrointestinal Symptoms [0] Good Appetite [1] Poor Appetite/Nausea [2] Moderate Nausea/Vomiting [3] Severe. Incapacitating Nausea and Vomiting Score = 3. Fatigue and/or weakness [0] Not Tired or Weak [1] Mild Fatigue/Weakness
  85. 85. Thank You! Dr. Trevor Day Associate Professor of Physiology Department of Biology Faculty of Science and Technology Mount Royal University tday@mtroyal.ca For additional information on the solutions presented in this webinar please visit www.adinstruments.com

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