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NB: The review presented in this chapter and the start of the machine methodology presented at the beginning of the next chapterformed the basis of the publication: Byrnes CA, Bush A and Shineboume EA "Methods to Measure Expiratory Nitric Ortde in People" in "A Volume of Methods in Enqmology" edited by Lester Packeter Academic press Inc. 1996. 109
5.1 Chapter 5: Methodological assessment of the chemiluminescence analyser Introduction The previous chapter reviewed the methods available to measure NO in biological systems and determined that the most appropriate method for measuring NO in exhaled air using a chemiluminescence analyser, originally developed to assess NO pollution levels. The technical aspects of the chemiluminescence analysers were reviewed and the direct correlation between the intensity of light measured and the concentration of NO described. The instrument that I ultimately used was the Dasibi Environmental Corporation 'Model 2107'.In this chapter the characteristics of this analyser will be reviewed, as well as the other equipment. Connections between a number of analysers were developed to enable simultaneous measurement of NO, COz, flow and pressure. I will also mention the other people involved in the work and the role each person undertook. 5.2 The equipment and personnel 5.2.1 Chemilurninescence Analyser'Model 2107' The chemiluminescence analyser Model 2107 (Dasibi Environmental Corporation, Glendale, California, USA; local supplier Quantitech Ltd, Unit 3 Wolverton Rd, Old Wolverton, Mlton Keynes, UK) was designed for measuring NO concentrations within a 2-4000pbb range in a continuous ambient air sample and operated in ambient temperatures of 5400 C and humidity of O-9SVo.It had been adapted for online recording with a sampling rate at 240 mls/min and had a highly stabilised measurement capability (drift without auto zero over 24 hours of lppb). The original response time of this machine was long at 90 seconds, too long for the purpose for which it was now required. Modification of the circuitry and the sending of the pre-computer analogue signal directly to the chart recorder decreased the response time to 6.4 seconds (see below). The estimates of NO concentrations were decreased by lo-l1Vo at IOOVo humidity; however NO pre-drying of the expirate was not thought to be necessary (this may have been considered differently if the experiments were conducted in areas of high humidity such as within New Zealand). I did use a water absorber when doing the reservoir set of experiments only (see Section 6.4). This machine operated on the same principles as described in the last chapter. The gaseous specimen was drawn by the vacuum pump into a reaction chamber where the ozone was generated internally by an electrical discharger. The NO and 03 wore mixed in front of a red-sensitive photomultiplier tube, the light emission of this reaction was measured at 660 to 900nm in the red sensitive range, and the signal 110
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UNIVERSITV OF AUCKI'AND Abstract -
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There was no significant difference
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This is Dedicated: Dedication To th
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New Zealand The Paediatric Departme
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Contents Abstract ........tr Dedica
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6.5.4 The subjects............... .
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XIV
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List of Figures Figure l.l: Top l0
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List of Abbreviations ABPA allergic
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LADA N*N* dimethyl L-arginine LFA1
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t}ryem(eNoS)Typeltrendothelialnifti
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area, some of it has been expanded
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In addition, both of these groups a
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studies conducted throughout the Un
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Video clips of infants with a varie
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the Paediatric Society of New 7*ala
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inhaled anti-cholinergic agents and
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ecommended to monitor asthma at hom
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improvement to 70Vo PEF would have
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Furthermore, investigators have als
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poorer predictor of a diagnosis of
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Bronsky et al. 1998), and a long-te
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SIGN guideline (SIGN 2005) stated "
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In brief, the human airway can be d
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The ability to biopsy has led to st
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small proportion of cells recovered
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methodology of sputum induction and
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et al. 1999; Giannini, Di Franco et
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over a six month treatment prograrn
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There are other studies suggesting
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So is induced sputum in adults and
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asymptomatic patients independent o
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onchial hyper-responsiveness (Reich
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led to research which looked for le
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analyser machines. This thesis pres
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The pollution of the 'great smog of
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adults greater than 65 years (Ostro
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levels compared to those children l
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More recently 'Air Quality Indexes'
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Page 93 and 94: 3.1 Chapter 3: The synthesis, react
Page 95 and 96: BHa = tetrahyrobiopterin, FAD = fla
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Page 101 and 102: toxic products such as isoprostanes
Page 103 and 104: Figure 3.3: The nitric oxide syntha
Page 105 and 106: providing NO as a neurotransmitter
Page 107 and 108: The enzyme of this second pathway -
Page 109 and 110: inappropriate production from comme
Page 111 and 112: Nicotinamide adenosine dinucleotide
Page 113 and 114: enzyme and are competitively revers
Page 115 and 116: 4.1 Introduction Chapter 4: Methods
Page 117 and 118: 4.4 Nitrite and nitrate The measure
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Page 121 and 122: A group of instruments have been de
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Page 127 and 128: helium for 30 minutes to remove Oz.
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Page 135 and 136: wanted to compare the patterns of e
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Page 141 and 142: Delay time: the time between turnin
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Page 145 and 146: Chapter 6: Methodological studies o
Page 147 and 148: pulmonary circulation appeared to c
Page 149 and 150: al found levels of 10.3ppb compared
Page 151 and 152: Authors and Pap€r Sublect numbers
Page 153 and 154: only those measured by mouth or low
Page 155 and 156: 6. Check that the pens work. Fill w
Page 157 and 158: keys but it is currently in the mid
Page 159 and 160: Calibration pressures: the test por
Page 161 and 162: T-piece: tO The first figure shows
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Page 165 and 166: Figure 6.6: Mean exhaled NO levels
Page 167 and 168: NO and COz results from single exha
Page 169 and 170: 6.6.4 (iii) Comparison of exhaled n
Page 171 and 172: Figure 6.11b: COz levels at peak NO
Page 173 and 174: I also considered whether the diffe
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Page 177 and 178: need for methodological experiments
Page 179 and 180: The exhalations were carried out in
Page 181 and 182: Figure 7.2: Example of the tracing
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7.4.4 Results There was an increase
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Table 7.3: NO, COz and duration of
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The second set of exhalations invol
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Figure 7.6: Photographs of the chan
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Table 7.5: Exhaled NO results with
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the NO concentrations taken pre and
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Figure 7.10: Hyperbolic relationshi
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pressure over the likely range enco
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subjects with respiratory disease,
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8.1 Chapter 8: Exhaled nitric oxide
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standard error of the mean (SEM), s
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T-piece sampling system to analyser
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Figure 8.2a: Comparison of peak exh
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if yes who? if yes who? if yes who?
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There are limitations with regard t
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significantly between research grou
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controls. Kharitinov et al reported
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direct method in asthmatic children
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Figure 8.4a: The eftect of commenci
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had last used her p2 agonist inhale
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higher level of exhaled NO at 4.9pp
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9.1 Introduction Chapter 9: Exhaled
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For the oral measurements options i
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Table 9.1: The recommended standard
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taken from the available literature
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compartment correlated with the ser
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measurement with variable expirator
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to settle. We may have been measuri
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9.5 Off-line measurement NO determi
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(Hyde, Geigel et al. 1997; Tsoukias
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9.6 Nasal nitric oxide measurement
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measured over two 24 hour periods,
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women who coincidently experienced
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While these cross-sectional and the
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symptoms. Steroid response was sign
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'difficult asthma' also had higher
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season and then reduced down to bas
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of cross sectional studies with a t
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Ciabattoni et al. 2004; Petersen, A
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FEV9.5, symptom score and airways r
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Indomethacin - This medication alte
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89Vo for PCD, and above that exclud
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This low NO occurs despite higher t
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patients were followed through one
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differences noted based on filter,
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9.L6 Nitric oxide levels in exercis
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another study of 111 school childre
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Almost all studies used sedation, w
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peak exhaled NO production in the f
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flow and to minimize nasal contamin
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ecruit blood vessels in the lung. S
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Chapter 10: Reflections The outcome
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and I enrolled 52 asthmatic childre
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Edwards EA, Douglas C, Broome S, Je
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o Cass Byrnes & Nicky Holt. "Drugs
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o Byrnes CA. Paediatric fibreoptic
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Appendix 2: Questionnaire for enrol
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Al-Ali, M. K. and P. H. Howarth (20
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Archer, S. L., P. J. Shultz, et al.
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Baraldi, E., N. M. Azzolin, et al.
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Belda, J., P. Hussack, et al. (2001
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Bongers, T. and B. R. ODriscoll (20
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Brown, G. C. and C. E. Cooper (1994
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Castro-Rodriguez, J. A., C. J. Holb
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Cockcroft, D. W., D. N. Killian, et
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de Blic, J., V. Marchac, et al. (20
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Donaldson, S. H., W. D. Bennett, et
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Emi-Miwa, M., A. Okitani, et al. (1
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Frey, U., J. Stocks, et al. (2000).
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Gershman, N. H., H. Liu, et al. (19
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Grasemann, H., E. Michler, et al. (
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Hashimoto, T., K. Minoguchi, et al.
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Holgate, S. T., D. E. Davies, et al
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Iriso, R., P. M. Mudido, et al. (20
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Jones, A. W., M. Fransson, et al. (
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Kercsmar, C. M. (2006). Wheezing in
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Kleinert, H., T. Wallerath, et al.
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Lamblin, C., P. Gosset, et al. (199
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Li, X. and J. W. Wilson (1997). "In
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Lymar, S. V. and J. K. Hurst (1996)
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Massaro, A. F., S. Mehta, et al. (1
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Miyabara, Y., R. Yanagisawa, et al.
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contribute to impaired endothelium-
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ODell, T. J., R. D. Hawkins, et al.
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Panza, J. A., C. E. Garcia, et al.
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Peters, S. P. (2006). "Safety of in
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Prieto, L., L. Bruno, et al. (2003)
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Ribbons, K. A., X. J. Zhang, et al.
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Sacco, O., R. Sale, et al. (2003).
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Sessa, W. C., K. Pritchard, et al.
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Silvestri, M., F. Sabatini, et al.
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Stamler, J. S. (1994). "Redox signa
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Stuehr, D. J., N. S. Kwon, et al. (
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Tierney, W. M., J. F. Roesner, et a
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Upchurch, G. R., G. N. Welch, et al
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Wadsworth, R., E. Stankevicius, et
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Williams, O., R. Y. Bhat, et al. (2
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Yates, D. H., S. A. Kharitonov, et
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