On the Physiology of Hydrogen Diving and Its Implication for Hydrogen
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On the Physiology of Hydrogen Diving and Its Implication for Hydrogen Biochemical Decompression by Aiidreas Fahlman B.Sc. Hawaii Pacific University, 1996 A thesis submitted to the Faculty of Gnduate Studies and Research in partial fulfillment of the requirements for the degree of Doctor of Philosophy Department of Biology Carleton University Ottawa, Ontario Canada Aupst, 2000 @copyright 2000, Andreas Fahlman National Library Bibliothèque nationale ($1 of Canada du Canada Acquisitions and Acquisitions et Bibliographie Services services bibliographiques 395 Wellington Street 395. rue Wdlingîm Ottawa ON K1A ON4 Onawa ON KIA ON4 Canada Canada The author has granted a non- L'auteur a accordé une licence non exclusive licence allowing the exclusive permettant à la National Library of Canada to Bibliothèque nationale du Canada de reproduce, loan, dism%uteor seiî reproduire, prêter, distribuer ou copies of this thesis in microform, vendre des copies de cette thèse sous paper or electronic formats. la forme de microfiche/film, de reproduction sur papier ou sur format électronique. The author retains ownership of the L'auteur conserve la propriété du copy~@htin this thesis. Neither the droit d'auteur qui protège cette thèse. thesis nor substantial extracts fiom it Ni la thèse ni des extraits substantiels may be printed or otherwise de celle-ci ne doivent être imprimés reproduced without the authot's ou autrement reproduits sans son permission. autorisation. Abstract: Biochemical decompression, a novel approach for decreasing decompression sickness (DCS)nsk by increasing the tissue washout rate of the inert gas, was tested in pigs during simulated H2 dives. Since there is only limited physioiogical data on the use of Hz as a diving gas, direct calorimetry and respirometry were used to determine whether physiological responses to hyperbaric H2 and He are different in guinea pigs. The data suggested that responses in hyperbaric heliox and hydrox cannot be expiained solely by the thermal properties of the two gas mixtures. To increase the washout rate of Hz, a Hz-metabolizing microbe (Mefhanobrevibacier smithii) was tested that converts Hz to H20 and Ch.Using pigs (Sus scrofa) cornparisons were made between untreated controls, saline-injected controls, and animals injected with M. smithii into the large intestine. To simulate a Hz dive, pigs were placed in a dry hyperbaric chamber and compressed to different pressures (22.3-25.7 atm) for times of 30-1440 min. Subsequently, pigs were decompressed to Il atm at varying rates (0.45-1.80 atm min-'), and observed for severe syrnptoms of DCS for 1 h. Chamber gases (O2,Nt, He, Hz,Ch) were monitored using gas chromatography throughout the dive. Release of Ch in untreated pigs indicated that Hz was being metabolized by native intestinal microbes and results indicated that native Hz-metabolizing microbes may provide some protection against DCS following hyperbaric Hz exposure. M. smithii injection further enhanced CH4 output and lowered DCS incidence. A probabilistic model estimated the effect of H2-metabolism on the probability of DCS, P@CS), after hyperbaric Hz exposure. The data set included varying compression and decompression sequences for controls and animals with intestinal injections of H2 metabolizing microbes. The model showed that increasing total activity of M. smithii injected into the animals reduced P@CS). Reducing the tissue concentration of the inert gas significantly reduced the nsk of DCS in pigs, Mersupporting the hypothesis that DCS is pnmarily caused by elevated tissue inert gas tension. The data provide promising support for the development of biochemical decompression as an aid to human diving. Acknowledgements: First and foremost, 1 would like to thank Dr. Susan R. Kayar for shaping my scientific career and having patience with my endless Stream of questions. Also, a special thanks to Dr. Kenneth B. Storey who has had patience with me throughout the yean at Carleton University, and taught me the painhil lesson about Occam's razor. 1 would also like to thank Jan Storey and Diana Temple for their editorial help with al1 my material. I would like to thank my cornmittee members, Dr. Steven Brooks and Dr. Thomas Moon for their helpful suggestions. A special thanks to al1 the personnel at the Naval Medical Research Center in Bethesda, Maryland, for al1 their help and encouragement. 1 am particularly grateful to the Hz-diving team of Mr. Ayers, Mr. Rarnsey, Mr. Ruopoli, and Mr. Morris without whose help none of this work would have been possible. Many thanks to Colonel McNamee who taught me to do surgery on pigs and the staff at LAMSD for al1 the help with surgeries and animal care. In addition, thanks to Drs. leff Hirnm, Paul Weathersby, Charles McClure, and Peter Tikuisis for al1 their mathematical and computational help. A couple of people to whom I owe the inspiration for this work are Dr. Varis Gnindmanis, Dr. John Culliney, Dr. Bulen Terem, Dr. Daniel Gefioh, and Mr. Brian Quinn. Sist men inte minst, tusen tack till min familj Som har stott mig hela vagen, speciellt min mor och mina morforaldrar and to Angie for love and support. Table of Contents Page Title Page I Acceptance Sheet i i Abstract iii Acknowledgements v Table of Contents vi List of Abbreviations vii List of Tables xi List of Figures xiv List of Appendices xvi C hapter One: General Introduction 1 Chapter Two: Calorimetry and respirometry in guinea pigs in hydrox 14 and heliox at 10-60 atm. Chapter Three: Calibration of the physical properties of the hyperbaric 45 chamber for diable estimation of Chrelease rate (;CH, ) Chapter Four: Reduction in decompression sickness risk by native 66 Hz-metabolizing intestinal microbes in pigs &er Hzhyperbaric exposure. Chapter Five: Injection of HI-rnetaboiizing microbes into the intestines 96 of pigs reduces decompression sickness risk &er hyperbaric exposure to Hz. Chapter Six: On the likelihood of decompression sickness during H2 bioc hemical decompression. Chapter Seven: General Discussion Publication List Re ferences List of Abbreviatioos %O - part per thousand A - parameter for BUG atm - atmosphere absolute, pressure unit equal to 1 .O 1 bar or 0.1 MPa BUG - variable for INJ or ;CH, C - control CH4 - methane c[CH4] - CH4 concentration in the charnber during the final hour at constant pressure d[CH4] - CH4 concentration in the chamber during decornpression CO2 - carbon dioxide DCS - Decompression Sickness G - scaling factor h, - heat tram fer coefficient H2 - hydrogen He - helium HPNS - high pressure nervous syndrome MJ - total H2metabolizing activity injected M - metabolic heat production rate N2- nitrogen O2- oxygen p - probability ppm - part per million P - pressure, measured in atm vii P(DCS) - Probability of DCS Pain- alveolar partial pressure Pmb - ambient pressure Pblood- arterial inert gas tension PH20 - water vapour pressure PO2 - partial pressure of Ol PH2 - partial pressure of Hz PHe - partial pressure of He PFC - perfluorocarbon Ptis- tissue tension PtisH2- Hydrogen tissue tension PtisHe- Helium tissue tension q - the rate at which gas [x] is added @pm min") Qc - heat losses by convection across body surfaces QcD - heat loss by conduction Qz - total heat loss rate QEVAP- heat loss by evaporation QR- heat loss by radiation QRC- heat loss by convection by respiration QRE- heat loss due to evaporation from the respiratory tract QSE- heat loss due to evaporation fiom the skin O D - dilution factor r - risk fûnction viii S - storage of body heat SC - surgical control STP - standard temperature (O°C) and pressure (1 atm) T - animal injected with Hzmetabolizing microbes t - time T - time constant T 1 - last time at which diver was definitely fiee of signs of DCS T2 - tirne at which definite signs of DCS occurred Tmb - ambient temperature TCH- chamber temperature Thr - ïhreshold parameter Tend - time for end of observation penod UC - untreated control V - volume v - flow rate V CH, - methane release rate c ;CH, - Chrelease rate in chamber during compression d ;CH, - Chrelease rate during decornpression Ic CH, - mean Chrelease rate during the last three hours at constant pressure tc CH, - rnean CH( release rate during the first three hours at constant pressure V 0 - oxygen consumption rate Wt - weight Z - transformation of fraction towards new steady state List of Tables Table Page Narne of common gas mixtures used for diving, their composition, and approximate pressures they are used at. Mean (f 1 stdev) oxygen consumption rate ( O?), mean total heat loss rate (Qt),and average calorimeter temperature (T,), for guinea pigs in heliox (n = 6) and hydrox (n = 6) at 10-60 atm, and in air (n = 16) at 1 atm, as measured by respirometry and calorimetry. Mean (f 1 stdev) of average heat loss rate (Pr),oxygen consurnption rate (c O,), and dorimeter temperature (Ta) for guinea pigs (n = 5) in air at 1 atm during a 6 h enclosure in a calorimeter. Mean (I1 stdev) heat loss rate (Qs)fiom direct calonmetry (actual), estimated by a mathematical model (Eq. 2, theoretical), and percent Qr partitioned into various avenues of heat loss as a percentage of Qr (Eq. 2). Pressure. gas mixture, and percent difference in total heat loss rate (Qr) (hydroxheliox) for a test to compare the difference in Qr between heliox and hydrox based soleiy on their differing thermal properties (TP 43, versus their measured values. Sensitivity test of model to estimate total heat loss rate (Qr) using Models 0, 1 .O, and 2.0 to estimate Qr for 1 atm air, heliox and hydrox, respectively . Flow meter calibration.