ContentServer 3

Respirology (2008) 13, (Suppl.
2) A25A74
doi: 10.1111/j.1440-1843.2008.01252.x
TSANZ Poster Sessions

Asthma & Allergy SIG: Poster Session 1.
Basic Science
TP 001
EARLY LIFE PNEUMOVIRAL INFECTION INDUCES THE

DEVELOPMENT OF PULMONARY ANGIOGENESIS
TP 002
ANTIGEN-SPECIFIC TH1 CELLS INDUCE AIRWAY

HYPER-REACTIVITY IN CO-OPERATION WITH
LIPOPOLYSACCHARIDE IN BALB/C MICE
NICOLE HANSBRO1, SARAH BOUSTANY2, BRIAN OLIVER2, JANETTE

BURGESS2, JUDY BLACK2, HELENE ROSENBERG3, PHIL HANSBRO1,
PAUL FOSTER1
1
Centre for Asthma & Respiratory Disease, University of Newcastle, NSW,
2300, 2Department of Pharmacology, University of Sydney, NSW, 2006, and
3
Laboratory of Allergic Diseases, National Institute of Allergy and Infectious
Diseases, National Institutes of Health, Bethesda, Maryland, USA
Early life infection with RSV is associated with development and exacerbations
of asthma. Airway vascularity and pro-angiogenic factors (VEGF, bFGF, angiogenins) are upregulated in the airways of asthmatics. We investigated whether
early life pneumoviral infection leads to the development of pulmonary angiogenesis, and whether this is due to viral-induced up-regulation of proangiogenic factors in the lung.
Methods Lung tissues were collected following primary or secondary infection of newborn BALB/c mice with pneumonia virus of mice (PVM). Viral titre
and expression levels of pro-angiogenic factors in lungs were determined by
qPCR. Angiogenesis was quantified by immunohistochemistry using the angiogenic markers CD31 (PCAM-1) and von Willebrand Factor (vWF).
Results Angiogenesis was apparent 14 days post infection (dpi)
(CD31 = 0.3 0.1, vWF = 0.2 0.2), peaked at 28 dpi (CD31 = 30.8 3.6,
vWF = 38.4 5.3) and regressed by 35 dpi (CD31 = 0.3 0.3, vWF = 0). A
secondary infection at day 82 re-activated angiogenesis by day 96
(vWF = 46.8 6.7). Detection of pro-angiogenic factors (VEGF, Ang1 and
Tie2) in the lung correlated with the observed angiogenesis.
Conclusions Early life pneumoviral infection induces the development of
angiogenesis which is spatially associated with the airways. This model is the
first to demonstrate a specific link between early life respiratory viral infection
and the induction of angiogenesis in the lung.
Supported by The Hunter Medical Research Institute, CRC for Asthma and
Airways and NH&MRC.
Conflict of Interest No.
M YANG, C MA, PS FOSTER

Priority Research Centre for Asthma & Respiratory Diseases, Faculty of
Health, University of Newcastle, Newcastle NSW 2300
A Th1 response has been considered to be protective for asthma, on the basis
that it might antagonize the dominant Th2 response. However, there is increasing evidence that under certain circumstances, Th1 cells are involved in the
induction of asthmatic manifestations and can potentiate pathological
responses. The mechanisms by which Th1 cells are involved in asthma remain
unclear. We assessed the interaction between Th1 cells and lipopolysaccharide (LPS) in the induction of airway hyper-reactivity (AHR).
Methods Antigen-specific DO11.10 Th1 or Th2 cells were generated in vitro,
in the presence of OVA323359 peptide with IL-12/anti-IL-4 or IL-4/anti-IFNg
respectively. Antigen-specific Th1 or Th2 cells (5106) were injected intravenously and followed by intranasal challenge with 10 mg OVA323359 daily for 4
consecutive days. Some groups of animals that received Th1 cells were administered 50 ng LPS together with the last OVA323359 challenge. Other groups
were treated with anti-IFNg, dexamethasone or cytotoxic antibody to deplete
neutrophils.
Results Antigen-specific Th1 cells alone did not induce airway inflammation
or AHR in mice challenged with OVA323359. Additional treatment with LPS
resulted in significantly increased airway reactivity and neutrophil infiltration
into the lung. Neutralization studies showed that the interplay of LPS and IFNg
underpinned this response, which was steroid-resistant. Neutrophils recruited
into the lung were not responsible for induction of AHR.
Conclusions Our investigation establishes that an interaction between
antigen-specific Th1 cells and LPS is required to induce AHR in this experimental model. Importantly, this AHR is steroid-resistant and is critically dependent on IFNg.
Supported by University of Newcastle, CRC and NHMRC.
Nomination APSR Early Career Development Award.
TP 003
ABSENCE OF SPHINGOSINE KINASE 1 LEAD TO

REMODELLING AND INCREASED RESPONSIVENESS OF
PULMONARY VASCULATURE IN CHRONIC AIRWAY
INFLAMMATION
RAINER V HABERBERGER1, CHRISTOPH TABELING3, SUE RUNCIMAN1,
RICHARD L PROIA2, NORBERT SUTTORP3, IAN GIBBINS1,
MARTIN WITZENRATH3
1
Dept. of Anatomy & Histology, Flinders University, Adelaide, SA 5001,
2
NIDDK, National Institutes of Health, Bethesda, USA, and 3Dept of Internal
Medicine, Charite, Universittsmedizin Berlin, Germany
Rationale Inflammatory mechanisms play a prominent role in the pathogenesis of pulmonary hypertension. The bioactive sphingolipid sphingosine
1-phosphate (S1P) is an important vascular modulator.
Methods Mice deficient in one of the two S1P synthesizing sphingosine
kinase (SK) isoforms, SK1, and the corresponding wild-type mice (wt, C57/Bl6)
were employed. Allergen-evoked airway inflammation was induced by longterm recurrent ovalbumin-exposure and the effects of SK1-deficiency on lung
vasculature were investigated.
Results Under inflammatory conditions, quantitative RT-PCR showed
upregulated pulmonary levels of eotaxin mRNA in SK1-KO as compared to
nave mice, whereas mRNA-expression of IL-4 as well as S1P-synthesizing
and degrading enzymes was unaltered. BAL of chronically inflamed SK1deficient mice contained more cells in total and a larger number of CD68positive macrophages/monocytes as compared to BAL of wt lungs. Following
allergen-exposure, a remarkable remodelling of intraacinar vessels and small
pulmonary arteries with longitudinally orientated smooth muscle cells luminal to
the internal elastic membrane was observed in SK1-KO mice as compared to
wt mice. Using isolated ventilated mouse lungs perfused with the thromboxane
receptor agonist U46619, vascular hyperresponsiveness was observed in
allergen-exposed lungs from wt and SK1-KO mice, with higher vascular
responsiveness of SK1-KO lungs.
Conclusion Absence of SK1 increased alterations in morphology and function of pulmonary arteries under chronic inflammatory conditions suggesting a
possible role of the SK1/S1P system in the pathobiology of inflammationassociated pulmonary arterial hypertension.
Journal compilation 2008 Asian Pacific Society of Respirology
TP 004
STREPTOCOCCUS PNEUMONIAE VACCINE PREVENAR:

A POTENTIAL THERAPY FOR THE SUPPRESSION OF ASTHMA
ALISON N THORBURN, PAUL S FOSTER, PETER G GIBSON,
PHILIP M HANSBRO
Priority Research Centre for Asthma and Respiratory Disease and Hunter
Medical Research Institute, University of Newcastle, NSW 2300 and John
Hunter Hospital, NSW 2305, Australia
Recently, it has been shown that Streptococcus pneumoniae (Spn) respiratory
infection may inhibit the development of inflammatory responses associated
with allergic lung diseases such as asthma. In this study we investigated the
potential of current human Spn-based vaccines to suppress ovalbumin (OVA)induced allergic airways disease (AAD) using mouse models.
Methods AAD was induced in 68 wk old female BALB/c mice by intraperitoneal sensitization with Th2-inducing adjvant (alum) on day 0 and intranasal
challenge with OVA on day 1215. AAD was assessed on day 16. Mice were
administered with Prevenar or Pneumovax with and without Th1-inducing
adjuvant (CpG) at the time of OVA-sensitization. The effects of Spn vaccines
on the development of hallmark features of AAD were determined.
Results Prevenar significantly decreased eosinophil influx into the lung and
airways hyperresponsiveness compared to untreated allergic controls. Furthermore lymph node cell cultures stimulated with OVA showed a significant
decrease in Th2 cytokine release. Pneumovax had no effect on AAD. CpGs in
combination with vaccines had no additional effect.
Conclusions Prevenar may have potential applications as a therapy for the
suppression of allergic lung disease such as asthma.
Support Asthma NSW, HMRI, Asthma CRC.
Conflict of Interest This work is the subject of a patent application.
A26
Respirology (2008) 13, (Suppl. 2)
TP 005
A SHEEP ASTHMA MODEL FOR COMPARATIVE STUDIES ON

THE FUNCTION OF AIRWAY SMOOTH MUSCLE
ROB BISCHOF1, KEN SNIBSON2, STUART HIRST1, E MEEUSEN1
Department of Physiology, Monash University, Clayton, VIC 3800, and
2
Centre for Animal Biotechnology, The University of Melbourne, Parkville,
VIC 3010
The underlying biological processes involved in airway remodelling are still

poorly understood and scientific progress in this area has been restricted by
the lack of a suitable experimental model. Sheep have been widely used as
models for lung development and pulmonary diseases including asthma. We
have developed an experimental sheep model of human asthma based on
house dust mite (HDM) that displays many of the hallmark features of the
human disease including raised IgE levels, eosinophilia and increased airway
mucus production. Repeated (chronic) allergen challenges in this model result
in airway tissue remodelling with increased airway smooth muscle (ASM) and
collagen, and deteriorating lung function.
In light of the inherent difficulties in obtaining ASM from human lung biopsies,
the sheep HDM asthma model provides a platform for relevant functional
studies of ASM under different experimental conditions. Ovine ASM cultures
from the trachealis muscle were established by explant culture or following
enzymatic dissociation, and cells examined by fluorescence microscopy and
flow cytometry at passages 25. Cultured ovine ASM cells showed differential
serum growth responses and displayed typical morphological features of
smooth muscle cells, with positive immunoreactivity for smooth muscle-specific
a-actin and desmin. Preliminary comparative studies of the phenotypic and
functional properties of ASM at different airway levels from asthmatic and
non-asthmatic animals are underway.
These studies highlight the potential for this highly relevant translational model
to study changes in ASM while monitoring the kinetics of the asthmatic
response and, given the longevity of sheep, its use as a tool for assessing
chronic disease and long-term therapeutic procedures.
Supported by the NHMRC.
TP 006
RELAXIN TREATMENT REVERSES AIRWAY REMODELLING

IN MURINE ALLERGIC AIRWAYS DISEASE
SIMON ROYCE1, YU MIAO1, MELISSA LEE1, CHRISHAN SAMUEL2, MIMI TANG1
Dept. Allergy and Immunology, Murdoch Childrens Research Institute,
The Royal Childrens Hospital, and 2Howard Florey Institute, Victoria 3052
Airway remodelling is a set of structural changes in asthma that has been

suggested as a novel target for the treatment of severe and persistent asthma.
Mice deficient in the antifibrotic hormone relaxin develop age-related changes
similar to airway remodelling, including severe bronchial fibrosis. We used an
established murine model of chronic allergic airways disease to examine the
effect of treatment with exogenous relaxin on airway remodelling and airway
hyperresponsiveness.
Methods A chronic ovalbumin allergic airways disease model was induced in
8 week old female Balb/c mice. Following the ovalbumin aerosol challenge
period, mice were treated with recombinant H2 relaxin (0.8 mg/ml) or vehicle
control via s.c. mini-osmotic pumps. Two weeks later airway hyperresponsiveness was assessed, tissues collected and airway inflammatory and remodelling parameters measured.
Results Recombinant relaxin administered systemically was able to reverse
collagen deposition around the airways to levels seen in saline control animals.
This resulted in reduction in airway hyperresponsiveness measured by invasive plethysmography. Relaxin-treated mice did not exhibit changes in bronchoalveolar fluid inflammatory cells or in goblet cell hyperplasia compared to
untreated ovalbumin animals. Matrix metalloprotease levels were increased, a
likely mechanism for the antifibrotic effect.
Conclusions Relaxin was able to dramatically reduce airway remodelling in
a chronic mouse model of allergic airways disease. Relaxin treatment also
resulted in reduced airway hyperresponsiveness. Further trials are warranted
to investigate the efficacy of relaxin in the treatment of allergic airways disease.
Supported by the ARC.
TP 008
TP 007
A MOUSE MODEL OF SUBLINGUAL ALLERGEN

IMMUNOTHERAPY WHICH INHIBITS EXPERIMENTAL ASTHMA
MEASURING SEVERE LUNG DISEASE IN MICE

A CAUTIONARY TALE
CHARLES L HARDY, JUN YAO, JENNIFER M ROLLAND, ROBYN E OHEHIR

Department of Immunology, Monash University, Melbourne, Australia;
Department of Allergy, Immunology and Respiratory Medicine, The Alfred
Hospital; CRC for Asthma & Airways, Sydney, Australia
ALEXANDER N LARCOMBE1, GRAEME R ZOSKY1, DEBRA J TURNER1,

ZOLTAN HANTOS2, PETER D SLY1
1
Telethon Institute for Child Health Research, Centre for Child Health
Research, University of Western Australia, Perth, Western Australia, 6008,
and 2Department of Medical Informatics and Engineering, University of
Szeged, H-6720 Szeged, Hungary
Allergen-specific immunotherapy (SIT) via the subcutaneous route is an established treatment for respiratory allergies but is associated with severe systemic
reactions. Sublingual allergen immunotherapy (SLIT) is a promising alternative
to SIT with improved safety. However, the immune mechanisms have not been
delineated and there is therefore a need for experimental models. We compared efficacy of SLIT with whole allergen protein versus allergen immunodominant peptide in an experimental asthma model.
Methods BALB/c mice received the model allergen ovalbumin (OVA; OVASLIT mice) or OVA323339 immunodominant peptide sublingually prior to intraperitoneal sensitization with OVA in alum. Mice were challenged intranasally
with OVA, or fluorescent OVA to identify APC which had endocytosed allergen,
and immunological parameters analysed 24 hours later.
Results SLIT-OVA mice had markedly decreased airway eosinophilia, frequency of draining lymph node (LN) cells producing the Th2 cytokines IL-4, IL-5
and IL-13, and concentrations of airway TGF-b and serum OVA-specific IgE.
SLIT with OVA323339 was less effective. Compared to OVA sensitized and
challenged mice, SLIT-OVA mice had altered proportions of allergen-laden
CD11c+CD11b+ lung dendritic cells (DC), and decreased DC expression of
MHCII and the co-stimulatory molecules CD40 and CD86 in lung and lungdraining LN.
Conclusions SLIT decreased key clinical asthma symptoms, associated
with altered pulmonary DC maturation and allergen uptake. Our model will be
useful for further defining SLIT mechanisms of action.
Background The flexiVent small animal ventilator (SCIREQ, Canada) allows

measurement of detailed respiratory mechanics on animals as small as 10g. It
uses the forced oscillation technique (FOT) to measure respiratory system
impedence (Zrs) by applying a signal containing 19 mutually prime sinusoidal
frequencies ranging from 0.25 to 19.625 Hz at the airway opening. The constant phase model (CPM) is fit to Zrs to calculate airway resistance (Raw) tissue
damping (G) and tissue elastance (H). The default settings of flexiVent apply an
unweighted (absolute) fitting of the CPM to the Zrs spectra. This may not be
appropriate for mice with severe lung disease.
Methods Adult female BALB/c mice with influenza-induced lung disease
were challenged with methacholine (MCh). Data were analysed using the
default primewave settings of flexiVent v5.1. These same data were then
adjusted to remove the two lowest frequencies (0.25 and 0.625 Hz) and a
relative (weighted) fitting of the CPM was applied.
Results The default analysis produced a poor model fit and uninterpretable
measurements of Raw, G and H, especially at higher MCh concentrations.
Applying the weighted fitting improved the application of the CPM to Zrs
spectra. A better partitioning of the CPM into airway and tissue compartments
was achieved by removing the lowest two frequencies from the analysis.
Conclusion The default flexiVent settings provided by SCIREQ are not suitable for accurate measurement of lung mechanics in mouse models of severe
lung disease.
Thoracic Society of Australia and New Zealand Annual Scientific Meeting 2008
TP 009
A27
TP 010
NEUTROPHIL INFLUX DURING CHLAMYDIAL LUNG INFECTION

DETERMINES THE PHENOTYPE OF ALLERGIC AIRWAYS
DISEASE
HAEMOPHILUS INFLUENZAE (Hi) INFECTION IN ASTHMA MAY

DRIVE THE DEVELOPMENT OF NEUTROPHILIC ASTHMA (NA)
JC HORVAT, KW BEAGLEY, PG GIBSON, PS FOSTER, PM HANSBRO

Priority Research Centre for Asthma and Respiratory Disease and Hunter
Medical Research Institute, University of Newcastle, NSW 2300 and John
Hunter Hospital, NSW, 2305, Australia
AMA-TAWIAH ESSILFIE, JULIE PRESTON, JAY HORVAT, MARGARET

DUNKLEY, PAUL S FOSTER, PETER G GIBSON, JODIE L SIMPSON,
PHIL M HANSBRO
Priority Research Centre for Asthma & Respiratory Disease, Hunter Medical
Research Institute, University of Newcastle, John Hunter Hospital,
Newcastle, Australia
Chlamydial lung infections are linked with asthma but the nature of the association is obscure. We have demonstrated that an ongoing, but not cleared,
chlamydial lung infection at the time of allergen sensitization promotes a skew
from a Th2 to a mixed Th1/Th2 allergen-specific T-cell response that promotes
neutrophilic and suppresses eosinophilic inflammation. During this neutrophildominated allergic airways disease (AAD), goblet cell hyperplasia and AHR are
reduced but still present. The resulting phenotype has features similar to
neutrophilic asthma in humans. We examined whether pulmonary neutrophil
influx during infection plays an important role in the ability of an ongoing
infection to induce AAD with this phenotype.
Mice were treated intra-peritoneally (IP) with both anti-keratinocyte chemokine
(aKC) monoclonal antibody (mAb) and anti-macrophage inflammatory
protein-2 (aMIP2) mAb, 3, 5, 7 and 9 days after inoculation with Chlamydia
muridarum (Cmu). This treatment regime resulted in decreased pulmonary, but
not systemic, neutrophilia during infection. Seven days after inoculation, mice
were sensitized to Ovalbumin (Ova) by IP injection. AAD was induced by daily
intranasal Ova challenges 1215 days after sensitization. On day 16, key
features of AAD were characterized and compared to non-antibody treated
controls.
Depletion of pulmonary neutrophilia during infection abrogated the Chlamydiamediated reduction in Ova-specific IL-5 release from T cells from the lung,
goblet cell hyperplasia and AHR during AAD. Furthermore infected treated
animals no longer mounted a robust pulmonary or systemic neutrophil
response upon the induction of AAD despite cessation of antibody treatment 7
days earlier.
Ongoing chlamydial respiratory infections substantially modify key allergenspecific immune responses in AAD with the composition of cellular inflammatory responses to infection crucial in determining the outcome of allergic
phenotype.
NA is characterized by the presence of neutrophilic lung inflammation. Hi

colonization has been shown in some people with NA, however it is not known
if colonization induces NA or if these patients are predisposed to colonization.
We investigated the nature of this association using mouse models of lung
infection and ovalbumin (OVA)-induced allergic airway disease (AAD).
Methods Mice were infected 10 days before (Before), the same day as
(During) or 10 days after (After) intraperitoneal OVA sensitization. AAD was
induced by intranasal OVA challenge 1215 days after sensitization, then
characterized on day 16. Lung-draining lymph nodes (LN) were restimulated
with killed Hi or OVA and IL-5, -13 and IFN-g release was determined. Eosinophils and neutrophils were enumerated in bronchoalveolar lavage and
airway hyper-responsiveness (AHR) was assessed by measurement
of airways resistance and dynamic compliance following methacholine
administration.
Results The induction of AAD after infection inhibited Hi clearance and
increased and decreased Hi-specific IL-5 and IFN-g, respectively. In all groups
infection decreased OVA-specific IL-5, -13 and IFN-g and eosinophil influx into
the lung but increased neutrophil recruitment and did not significantly alter
AHR.
Conclusion This combined model of AAD and Hi infection is consistent with
the clinical phenotype of NA and supports our hypothesis that Hi infection may
lead to the development of NA.
TP 012
NOVEL BRONCHODILATOR RESPONSE TO ROSIGLITAZONE

IN INTACT AIRWAYS IN MURINE LUNG SLICES
TP 011
PERSISTENCE OF AIRWAY HYPERRESPONSIVENESS IN MICE

INFECTED WITH INFLUENZA A AS INFANTS
EM BOZANICH1, R GUALANO2, AN LARCOMBE1, GR ZOSKY1, PD SLY1,
DJ TURNER1
1
Division of Clinical Sciences, Centre for Child Health Research, Perth, WA
6008, and 2Dept. of Pharmacology, University of Melbourne, Parkville, VIC
3052
Early life exposure to viral lower respiratory illness is thought to exacerbate the
development of allergic airways disease such as asthma. There are few published murine models of respiratory virus infection in infants and neonates. This
study aimed to characterize the inflammatory and physiological pattern of
Influenza A infection in infant mice.
Methods Infant BALB/c mice were inoculated on day 7 (d7) with 10mL
103.8 pfu Influenza virus A/Mem/1/71 (H3N1) or media control. Bronchoalveolar
lavage fluid was collected at d1-5, 7, 10, 14 & 21 post-inoculation to determine
the inflammatory profile. Following the establishment of peak inflammation and
clearance, physiology was assessed during the acute inflammatory period
(d5-post-inoculation) and resolution of infection (d21) using a small animal
ventilator and a modification of the forced oscillation technique. The Constant
Phase Model was fitted to Respiratory Impedance data to produce airway and
tissue mechanics. Airway hyperresponsiveness (AHR) was assessed to
inhaled methacholine (MCh) (0.0130 mg/mL) at d21.
Results Cellular inflammation peaked at d3 in Influenza infected mice
(1.23 106 (0.23) cells/mL) compared to control animals (2.79 105 (0.29)
cells/mL; p = 0.007) and had resolved to control levels by d14 (1.65 105
(0.48) cells/mL vs 1.53 105 (0.22) cells/mL; p = 0.837). Mice infected with
Influenza A virus showed changes to baseline mechanics at d5, and AHR to
inhaled MCh (p = 0.01) at d21 compared to control animals.
Conclusions These findings confirm that Influenza A virus induces significant changes in baseline mechanics during acute infection in infant mice, and
that AHR persists beyond the resolution of inflammation.
Supported by NHMRC.
Conflict of Interest None.
JANE WARD1, YAN BAI2, MICHAEL SANDERSON2

Dept of Pharmacology, University of Melbourne, Parkville, Victoria 3010,
and 2Dept of Physiology, University of Massachusetts Medical School,
Worcester, MA, USA
The ligand-activated transcription factor proliferator activated receptor gamma

(PPARg) has been implicated in both inflammatory and remodelling processes
in asthma. We have previously shown that the PPARg ligand rosiglitazone
(RGZ) reduces airways hyperresponsiveness (AHR) in a murine model of
allergen-induced inflammation without inhibiting inflammatory cell infiltration
[1]. The hypothesis that RGZ reduces AHR by directly causing airway smooth
muscle (ASM) relaxation was tested by measuring contractility of small airways
in mouse lung slices in which in situ organization is maintained.
Methods C57BL/6 mice were killed by sodium pentobarbitone overdose
(i.p.), the trachea cannulated and chest cavity opened. The lungs were inflated
with liquid agarose that solidifies to stiffen the tissue for sectioning (~130 mm)
[2]. The effects of RGZ (1100 mM) +/- PPARg antagonist (GW9662 1 mM) on
small airway contraction in response to a submaximal concentration of methacholine (MCh, 300 nM) were assessed. RGZ was also tested after pretreatment with ryanodine (50 mM) and caffeine (20 mM) to elicit a sustained
high [Ca2+]i within ASM. Under these conditions, MCh-induced contraction
occurs without a change in the [Ca2+]i via Ca2+ sensitization.
Results The MCh-induced contraction (40% reduction in lumen area) was
reversed in a concentration-dependent manner by RGZ. This acute relaxation
to RGZ (EC50 < 30 mM, complete relaxation 100 mM, n = 7) was not prevented
by GW9662. Following ryanodine-caffeine treatment, which elicited transient
airways contraction followed by relaxation, subsequent contractions to MCh
were also sensitive to RGZ.
Conclusions The novel bronchodilator action of RGZ in murine small
airways may be PPARg-independent. Its mechanism of this action in this
preparation is likely to involve inhibition of both MCh-induced increases in
[Ca2+]i and Ca2+ sensitization of ASM.
Funding Australian Lung Foundation
References
1. Ward, JE et al. (2006) Pulm. Pharm. Exp. Ther., 19: 3946.
2. Bergner, A and Sanderson, MJ (2002) J. Gen. Physiol. 119: 187198.
A28
TP 013
TP 014
THE RELATIONSHIP BETWEEN AIRWAY SIZE AND

SENSITIVITY TO CARBACHOL USING PORCINE AIRWAY
SEGMENTS
VASCULAR REMODELLING IN A SHEEP MODEL OF CHRONIC

ASTHMA
JESSICA KERMODE1,2, BRIAN OLIVER1,2,3, BRENT MCPARLAND1,2,3

1
Discipline of Pharmacology, University of Sydney, 2Respiratory Research
Group, and 3Woolcock Institute of Medical Research, NSW, Australia 2006
Asthmatic airways are highly sensitive to a number of stimuli. The reason

underlying this increased sensitivity is poorly understood. A previous study
found that sensitivity to acetylcholine relates to the diffusion distance of the
drug. We hypothesize that drug-diffusion distance would only influence drugsensitivity if the drug is metabolized while diffusing to its target. Therefore, the
aim of the present study was to investigate the relationship between diffusion
distance and sensitivity to carbachol (CCh), which acts on the same receptors
as ACh, but is not as easily metabolized.
Methods 22 airway segments (tubes) with a range of internal diameters (1.4
to 6.3 mm; surrogate for diffusion distance) were dissected from 13 female pig
lungs. Cumulative concentration curves were generated to carbachol applied to
the outside surface of tubes. Sensitivity was measured as the negative logconcentration of carbachol that would cause a half-maximal increase in luminal
pressure/response (-EC50). ID measurements were made from images of
stained cross sections (8 mm) cut from cryo-imbedded segments.
Results There was no significant correlation (n = 22, r ~ 0, p > 0.05) between
ID and -EC50 to CCh.
Conclusions Agonists need to be metabolically unstable, such as acetylcholine, for airway size to influence sensitivity. The smaller diffusion distance
between the luminal epithelium or mast cells and the airway smooth muscle
may in part account for the increased sensitivity in asthma.
It is now recognized that long-term structural and functional changes to lung

tissues (airway remodelling) in chronic asthmatics causes significant increases
in morbidity. We have previously shown that repeated challenges of House
Dust Mite (HDM) to sheep lungs results in significant remodelling of the
airways. We now assess remodelling of the bronchial vascular compartment of
the airways of sheep chronically challenged with HDM.
Methods Infusions of HDM were administered to specific lung segments of
allergic sheep, once weekly over a six month period. The equivalent segment
in the opposite lung was used as an untreated internal control. The bronchiolar
airway walls were assessed for vascularity by computer-assisted morphometric
analysis of Massons Trichrome stained histological sections.
Results Analysis of the morphometric data revealed that in some sheep
there were statistically significant increases in the number of blood vessels in
the bronchiolar airway walls in HDM challenged lung segments compared to
the untreated internal control lung segments (88% increase p < 0.05, 38%
increase p < 0.05). When assessed as a group, there was on average a 41%
greater number of vessels in bronchiolar walls of HDM treated lung segments
compared to the average number of vessels in untreated control lung segments (n = 4 sheep). The total vascular area in bronchiolar walls was on
average 54% greater in HDM segments compared with untreated control
segments (n = 4 sheep).
Conclusion These preliminary results show that the sheep model can be
useful for the study of the mechanisms underlying vascular remodelling in
chronic asthma.
J VAN DER VELDEN1, SJ HIRST2, KJ SNIBSON1

Centre for Animal Biotechnology, University of Melbourne, Parkville,
Victoria, Australia 3010, and 2Kings College London, MRC & Asthma UK
Centre in Allergic Mechanisms of Asthma, London SE1 9RT, UK
TP 015
MAST CELL DENSITIES IN ENDOBRONCHIAL BIOPSIES AND IN

SMALL AIRWAYS ARE RELATED
M CARROLL1, N CARROLL1, A BADDELEY2, HJG GUNDERSEN3,
A JAMES1
1
West Australian Sleep Disorders Research Institute, Nedlands, WA 6009,
2
School of Mathematics and Statistics, University of WA, Crawley, WA 6009,
and 3Stereological Research Laboratory, University of Aarhus, Denmark
Introduction It is unclear whether mast cell counts in endobronchial biopsy
specimens of proximal airways are related to mast cell counts in the small
airways. Aim To compare mast cell density in endobronchial biopsy sections
with mast cell density in transverse sections of small airways using stereological methods.
Method Lungs were obtained post-mortem from 10 subjects who died of
non-respiratory causes and fixed in inflation. Three-dimensional (3D) mast cell
densities (cells/mm3) were determined on endobronchial biopsies and transverse sections of small airways using an optical disector on 30mm thick histological sections, stained with human anti-mast cell tryptase.
Results Mean 3D mast cell density over the inner airway wall in biopsies was
significantly related to mean mast cell density over the total airway wall in the
small airways (r ~ 0.80, p < 0.01). A minimum of three biopsies per case was
required to demonstrate this relationship. Within relevant count areas, 3D mast
cell density was about 1.6 times higher in the small airways than in the
biopsies.
Conclusion These findings suggest that inter-subject comparisons of mean
mast cell density in the inner airway wall in endobronchial biopsies will tend to
reflect inter-subject comparisons of mean mast cell density over the total
airway wall in small airways. This is despite mast cell densities being generally
higher in the small airways. However, mean counts from at least three biopsies
per case are required to accurately represent inter-subject comparisons.
Supported by Merck, Sharp and Dohme and University of Western Australia.
TP 016
MAST CELL DENSITIES IN ENDOBRONCHIAL BIOPSIES AND IN

THE WHOLE LUNG ARE NOT RELATED
M CARROLL1, N CARROLL1, A BADDELEY2, HJG GUNDERSEN3,
A JAMES1
1
West Australian Sleep Disorders Research Institute, Nedlands, WA, 6009,
2
School of Mathematics and Statistics, University of WA, Crawley, WA, 6009,
and 3Stereological Research Laboratory, University of Aarhus, Denmark
Introduction Mast cell density in the inner airway wall on endobronchial
biopsies from proximal airways reflects inter-subject comparisons of mean
mast cell density in both the large airways and the small airways. However,
whether it is also related to mast cell density in the lung as a whole is unknown.
Aim To compare the density of mast cells in endobronchial biopsy sections
with the density of mast cells in the lung as a whole using stereological
methods.
Method Lungs were obtained post-mortem from 10 subjects who died of
non-respiratory causes and fixed in inflation. Mean three-dimensional (3D)
mast cell density (cells/mm3) in the inner airway wall in endobronchial biopsies
was determined using an optical disector on 30 mm thick histological sections
stained with human anti-mast cell tryptase. 3D mast cell density in the whole
lung was determined by estimating total mast cell number in the lung with the
fractionator technique and normalizing this against estimated tissue volume in
the lung.
Results 3D mast cell density in the lung as a whole was systematically lower
than that in biopsies but, on average, was of a similar order of magnitude (103
cells/mm3) to that in biopsies. Mast cell density in biopsies was not significantly
correlated with mast cell density in the lung as a whole (r = 0.10).
Conclusion These findings suggest that mast cell counts in biopsies from
proximal airways are not related to mast cell density in the lung as a whole.
Supported by Merck, Sharp and Dohme and University of Western Australia.
TP 017
AGEING ALTERS AIRWAY AND CIRCULATING NEUTROPHIL

FUNCTION
KJ Baines1,4, JL Simpson2,3,4, RJ Scott1, PG Gibson2,4
School of Biomedical Sciences, 2School of Medicine and Public Health,
3
NHMRC Centre for Respiratory and Sleep Medicine, The University of
Newcastle, NSW, Australia, and 4Department of Respiratory and Sleep
Medicine, Hunter Medical Research Institute, John Hunter Hospital, NSW,
Australia
A29
TP 018
INNATE IMMUNE RESPONSES OF AIRWAY NEUTROPHILS ARE

IMPAIRED IN NEUTROPHILIC ASTHMA
KJ BAINES1,4, JL SIMPSON2,3,4, RJ SCOTT1, PG GIBSON2,4
School of Biomedical Science, 2School of Medicine and Public Health,
3
NHMRC Centre for Respiratory and Sleep Medicine, The University of
Newcastle, NSW, Australia, and 4Department of Respiratory and Sleep
Medicine, Hunter Medical Research Institute, John Hunter Hospital, NSW,
Australia
Age is an important factor in the development of neutrophilic airway diseases

such as neutrophilic asthma and chronic obstructive pulmonary disease
(COPD). The immune response changes with age, however alterations to
neutrophil function that occur during healthy ageing are not well understood.
Aim To investigate age related changes in the innate immune responses of
airway and circulating neutrophils.
Methods Healthy subjects (n = 13) aged 26 to 77 years underwent sputum
induction and blood collection. Neutrophils were isolated from induced sputum
and peripheral blood samples using magnetic cell separation and CD16 microbeads. Cells were cultured in RPMI 1640 1% FCS with or without LPS stimulation (100ng/mL). IL-8, IL-1b, TNF-a, and total MMP-9 protein was measured
by ELISA, and relative gene expression of IL-8, IL-1b, TNF-a, TLR2 and TLR4
was measured by real-time PCR.
Results Participants were divided into 2 groups by the median age
(55 years). Airway neutrophils from older subjects (>55 years) released significantly more TNF-a protein and had significantly higher levels of mRNA for
TLR2, IL-8, IL-1b and TNF-a compared to younger subjects (p < 0.05). Resting
blood neutrophils from older subjects had significantly higher levels of MMP-9
release and TLR4 mRNA compared to younger subjects (p < 0.05). Blood
neutrophils from older subjects stimulated with the innate immune agonist LPS
released significantly less IL-1b and TNF-a protein compared to younger
subjects (p < 0.05).
Conclusion There are distinct alterations in neutrophil function that occur
during healthy ageing. Alterations in the innate immune response are characterized by an enhanced spontaneous activation of both airway and circulating
neutrophils, and an impaired response of circulating neutrophils to LPS stimulation.
Funded by The Asthma Foundation of NSW, CRC for Asthma and NHMR.
Neutrophils play a crucial role in innate immunity and contribute significantly to

airway inflammation in neutrophilic asthma. The mechanisms of neutrophilic
asthma remain unknown, however activation of the innate immune response is
implicated.
Aim To investigate the innate immune response of isolated airway neutrophils in subtypes of asthma.
Methods Participants with asthma (n = 17) and healthy controls (n = 11)
underwent sputum induction. Selected sputum samples were dispersed with
DTT and airway neutrophils were isolated using magnetic cell separation with
CD16 microbeads. Neutrophils were cultured in RPMI 1640 1% FCS for
24 hours. IL-8, and TNF-a protein was measured by ELISA, and relative gene
expression of IL-8, TNF-a, TLR4 and TLR2 was measured by real-time PCR.
Results Airway neutrophils isolated from subjects with neutrophilic asthma
released lower levels of IL-8 (p = 0.055) and significantly lower levels of TNF-a
(p = 0.0038) protein compared to healthy controls. Airway neutrophils isolated
from participants with neutrophilic asthma also had significantly decreased
levels of IL-8 mRNA (p = 0.03) and lower levels of TNF-a mRNA (p = 0.049).
This reduced cytokine release was linked with a decreased TLR4 and TLR2
gene expression, which was 4 and 2.9 fold down-regulated, respectively in
neutrophilic asthma compared to healthy controls.
Conclusion Airway neutrophil innate immune cytokine responses are
impaired in neutrophilic asthma compared to healthy controls and this is linked
with a decreased gene expression of TLR4 and TLR2. Dysfunction of airway
neutrophils in neutrophilic asthma would impact on susceptibility to and severity of airway infections.
Funded by The Asthma Foundation of NSW, CRC for Asthma and NHMRC.
TP 019
AIRWAY SMOOTH MUSCLE CELL VOLUME IS UNCHANGED IN

ASTHMA
J ELLIOT1, R JONES1, T MAUAD,2 M DOLHNIKOFF,2 M ABRAMSON,3
K MCKAY4, T BAI5, F GREEN6, A JAMES1,7
1
West Australian Sleep Disorders Research Institute, Nedlands, Western
Australia 6009, 2Sao Paulo University, Sao Paulo, Brazil, 3Monash
University, Melbourne, Australia, 4Childrens Hospital at Westmead, Sydney,
Australia, 5University of British Columbia, Vancouver, BC, Canada,
6
University of Calgary, Calgary, Alta, Canada, and 7Medicine and
Pharmacology, University of Western Australia
Increased airway smooth muscle (ASM) in asthma may be due to hyperplasia
or hypertrophy of ASM cells. The contribution of extracellular matrix (ECM)
within ASM bundles has not previously been accounted for when estimating
ASM cell volume.
Aim To estimate the mean ASM cell volume in ASM bundles in asthma.
Methods Post-mortem tissues from control subjects (C n = 9); nonfatal (NFA
n = 11) and fatal (FA n = 10) cases of asthma were studied. On 30 mm transverse airway sections stained with haematoxylin, the volume density (NV) of
ASM cell nuclei was estimated using an optical disector (1000). The mean cell
volume (VC = 1/NV) was calculated, correcting for the volume fraction of ASM
(fASM) within the ASM bundle (corrected VC = 1/(NV fASM)). fASM was estimated
on 0.5 mm thick sections of the same airway stained with Massons trichrome.
Basement membrane perimeter (Pbm) was used to indicate airway size.
Results Table shows results for large airways.
C
NFA
FA
Sex
Airway
Pbm
ASM/Pbm
M/F
6/3
5/6
5/5
n
23
24
25
mm
15 3
14 4
15 3
mm2
0.03 0.01
0.05 0.02*
0.08 0.03*
fASM
VC
0.48 0.9
0.51 0.9
0.49 0.7
(mm3)
2613 752
2305 699
2131 564
Mean SD. (one-way ANOVA) *p < 0.05 for C v FA, NFA v FA.
Conclusion These data suggest that although ASM area is increased in

asthma, mean ASM cell volume is unchanged. Therefore hyperplasia, not
hypertrophy, of ASM cells is present in both mild and severe asthma. These
results were similar for both large and small airways.
Supported by the NHMRC Australia (Grants #343601, #446800)
Nomination Nil.
Conflicts Nil.
TP 020
HUMAN LUNG MAST CELL PRODUCTS MODULATE AIRWAY

SMOOTH MUSCLE CELL SECRETION FROM DONORS WITH
ASTHMA
H ALKHOURI1, F HOLLINS3, LM MOIR1,2, Y ALRASHDAN1, CL ARMOUR1,
CE BRIGHTLING3, JM HUGHES1
1
Respiratory Research Group (Faculty of Pharmacy and Discipline of
Pharmacology), University of Sydney, NSW, 2 Woolcock Institute of Medical
Research, Sydney, NSW, and 3Department of Respiratory Medicine,
University Hospitals of Leicester, UK
Asthma is characterized by airway inflammation and remodelling which contribute to airway hyperresponsiveness and episodic airflow obstruction. Mast
cell (MC) densities are higher on the smooth muscle (ASM) in asthma so their
mediators may modulate other ASM functions as well as cause contraction.
Aim To investigate the effect of MC mediators on chemokine and extracellular matrix (ECM) production by ASM cells from donors with and without
asthma.
Methods MC were isolated from the resected lung samples of 6 patients,
resuspended at 106 cells/ml in DMEM + 10% FBS and stimulated with IgE/antiIgE. Supernatants (SN) were collected after 2 and 24 h and the MC lysed.
Sub-confluent ASM cells from 6 donors with and without asthma were serum
deprived for 72 h before MC SN/lysates were added in DMEM + 10%FBS for
48 h. IL-8 and eotaxin levels in all ASM SN and MC SN/lysates were measured
by ELISA. Fibronectin and collagen IV deposition was measured in situ by
immunoassay following ASM cell lysis.
Results In asthmatic and non-asthmatic ASM cells all MC SN and lysates
reduced eotaxin release by up to 47% and 58%, whereas the 02 h MC SN
significantly increased IL-8 release to 178 35.9% (p = 0.0339) and
169 49% (p = 0.0445) of the FBS control respectively. However, only nonasthmatic ASM cell IL-8 release was increased by the MC 224 h SN
(216 85%; p = 0.0421) and cell lysates (215 47%; p = 0.0421). The 02 h
MC SN also increased fibronectin deposition to 143 16% (p = 0.008) by
asthmatic ASM cells only. MC SN and lysates had no effect on collagen IV
deposition.
Conclusions Activated mast cell mediators differentially modulated chemokine and ECM secretion by ASM cells from donors with and without asthma.
Thus mast cells may modulate their own recruitment to the smooth muscle and
remodelling locally in the airways in asthma.
Supported by NHMRC.
A30
TP 021
TP 022
TNF-a AND IFN-g DIFFERENTIALLY REGULATE AIRWAY

SMOOTH MUSCLE CD40 AND OX40L EXPRESSION
A MODEL OF IgE SENSITIZATION TO STUDY RHINOVIRUS

INFECTION
DAVID I KRIMMER1, MATAKITAU LOSELI1, J MARGARET HUGHES2,

BRIAN GG OLIVER1,3, LYN M MOIR1,3, JANETTE K BURGESS1,3
1
Discipline of Pharmacology, 2Faculty of Pharmacy, University of Sydney,
NSW 2006, and 3Woolcock Institute of Medical Research, Camperdown,
NSW 2050
M LINDSAY1,2, B THOMAS1,2, H DAGHER1,2, J MEANGER1,2, R GHILDYAL2,

PG BARDIN1,2
1
Department of Respiratory Medicine, Monash University and Medical
Centre, VIC 3168, and 2Department of Medicine, MIMR, VIC 3168
Chronic airway inflammation in asthma is maintained by provision of survival

signals to CD4+ T cells. CD40 and OX40 Ligand (OX40L) are cell surface
molecules expressed on airway smooth muscle (ASM) that enhance CD4+ T
cell activation and survival. We examined the effect of the cytokines tumour
necrosis factor-alpha (TNF-a) and interferon-gamma (IFN-g) on CD40 and
OX40L expression.
Methods Cell-surface CD40 and OX40L expression on human ASM in
culture from asthmatic (n = 13) and non-asthmatic (n = 14) donors following
stimulation with TNF-a and IFN-g was measured using cell surface ELISA. In
some experiments, inhibitors of signalling pathways were also added.
Results TNF-a and IFN-g synergistically increased CD40 expression to a
greater extent than either cytokine alone. This phenomenon was greater on
asthmatic (n = 8) than non-asthmatic (n = 9) ASM (p < 0.01). In contrast, TNF-a
and IFN-g did not increase OX40L expression in comparison to TNF-a and
IFN-g alone. The CD40 (p < 0.001, Non asthmatic) and OX40L expression
(p < 0.01, Non asthmatic) induced following stimulation by TNF-a and IFN-g
was reduced by the addition of c-jun N terminal kinase (JNK) inhibitor
SP600125. CD40 (p < 0.05, non asthmatic) and OX40L (p < 0.05, non asthmatic) expression following TNF-a stimulation was increased by the addition of
the mitogen-activated protein kinase kinase (MMEK)-1 inhibitor PD98059.
Conclusion Asthmatic ASM can potentially modulate airway inflammation by
increasing expression of signalling molecules in response to cytokines. IFN-g
may activate mechanisms for decreasing OX40L expression which may
prevent maintenance of CD4+ T cell activation by ASM. We demonstrate a
possible role for JNK as a relevant signalling molecule in airway inflammation.
The technique of IgE passive sensitization reproduces IgE-related allergic

responses in vitro and studies have validated this technique for investigations
modelling allergic smooth muscle responses. There are no studies investigating effects of IgE sensitization on rhinovirus (RV) infection. We hypothesized
that RV infection is enhanced by IgE sensitization, a consequence of diminished early innate immune responses.
Methods Beas-2B epithelial cells and primary culture airway fibroblasts were
sensitized with IgE 24 h7 d prior to infection with RV16. Samples of tissue
culture supernatant and cell lysates were collected over a 12 h period after
infection for analysis. Viral replication was measured by real-time RT-qPCR
and viral titration and type I interferon mRNA by RT-qPCR. IgE receptor mRNA
expression was examined using RT-PCR.
Results Initial studies to establish the model used human serum high in IgE
(>1000 IU/ml), this yielded inconsistent results and it was found that purified
IgE (1000 IU/ml) provided more reliable responses. Sensitization was established after 24 h IgE incubation and was comparable with up to 7 d. RT-PCR
detected mRNA for the IgE low affinity receptor only after sensitization. Following RV16 infection, vRNA was increased after 24 h in IgE sensitized cells
(p < 0.05), but this effect varied noticeably between and within cell lines.
Cellular expression of IFN-b mRNA increased with viral infection but in cells
sensitized with IgE lower levels of expression were noted (p < 0.05).
Conclusions IgE passive sensitization enhanced RV replication in vitro but
the model is constrained by significant variability between and within cell lines.
The effect of sensitization on RV replication may occur through the low affinity
IgE receptor.
TP 023
MMP AND TIMP PRODUCTION BY CULTURED AIRWAY SMOOTH

MUSCLE CELLS FROM DONORS WITH ASTHMA
C LEVY1, H ALKHOURI1, Q GE2, CL ARMOUR1, JM HUGHES1
Respiratory Research Group, Faculty of Pharmacy, and 2Discipline of
Pharmacology, University of Sydney, NSW 2006
TP 024
CIRCULATING EOSINOPHIL COUNTS ARE HIGHER AFTER A

HIGH FAT MEAL THAN A LOW FAT MEAL
Activated mast cells (MC) are present in higher numbers on the airway smooth
muscle (ASM) in asthma compared with other inflammatory airway diseases.
Matrix metallo-proteinases (MMPs) cleave chemokines and alter chemokine
gradients by degrading the extracellular matrix and thus may modulate MC
migration to the ASM.
Aim To determine the levels of MMP-2, MMP-9 and their inhibitors, TIMP-1
and TIMP-2, secreted by ASM cells from donors with and without asthma.
Method Confluent ASM cells were washed, serum-starved for 48 h and then
stimulated with Th1 (IL-1, TNF and IFN) or Th2 (IL-1, IL-4 and IL-13) cytokines
or left unstimulated. After 4 and 24 h,the SN were collected. The relative
amount of pro and active forms of MMP-2 and MMP9 in SN were determined
by gelatine zymography. TIMP-1 and TIMP-2 levels in the SN were measured
by ELISA.
Results Pro- and active MMP- 9 were not detected. However, pro-MMP-2
levels were high in SN of ASM cells from donors with (195.6 47.2 % positive
control/105 cells) and without (226.5 49.2 % positive control/105 cells)
asthma. A trend to increased active MMP-2 production by ASM cells from
donors with (7.3 2.7 % positive control/105 cells, n = 9) compared to without
(2.9 0.7 % positive control/105 cells, n = 11) asthma after 24 h was not
significant (p = 0.101). TIMP-1 and TIMP2 levels respectively were high in the
SN of cells from donors with (69.4 19.6 and 21.3 4.7 ng/105cells, n = 5)
and without (57.3 13.7 and 16.6 3.5 ng/105 cells, n = 5) asthma. Th1 and
Th2 cytokine stimulation did not affect MMP or TIMP release.
Conclusions Th1 and Th2 cytokines did not regulate ASM cell production of
MMP-2, TIMP-1 and TIMP-2. Altered ASM MMP-2 activity is unlikely to play a
role in MC chemotaxis to ASM cells from donors with asthma in vitro or their
presence on the ASM in asthma.
Supported by NHMRC & Rebecca L Cooper Medical Research Foundation.
PETER BLACK, SUSANNE BRODIE, CLAIRE ARANDJUS, FAY SOMMERVILLE,

SARAH LEY
Department of Pharmacology & Clinical Pharmacology, University of
Auckland, Private Bag 92019, Auckland, New Zealand
There has been a marked increase in the prevalence of asthma and other
allergic diseases in the last few decades. One of the explanations for this is the
change in our diet. One of the characteristics of the Western diet is a high
intake of both saturated and polyunsaturated fat. This prompted us to compare
the effects of high fat and low fat meals on the numbers of circulating eosinophils and other leukocytes.
Methods We studied 12 volunteers who had allergic rhinitis and/or asthma
and a peripheral eosinophil count at baseline of 200 107/L. This was a
randomized, crossover trial with participants studied on two different days. On
each occasion they arrived fasting and after bloods were drawn consumed a
3000 calorie meal. One of the meals was high in saturated fat and refined
carbohydrate. The other meal was low in saturated fat and high in fruit and
fibre. Bloods were drawn postprandially every hour for five hours.
Results Eosinophil counts were highest in the early morning and fell over the
course of the day but the decrease was less with the high fat meal (p = 0.03).
Over the same period of time the increase in lymphocytes (p = 0.016) was
greater with the high fat meal. The high fat meal was also associated with
greater increases in triglycerides (p < 0.0001) and cholesterol (0.004).
Conclusions In atopic individuals a high fat meal was associated with higher
circulating numbers of eosinophils and lymphocytes than an isocaloric meal
that was low in fat. Further studies of the effect of dietary fat on eosinophilic
inflammation are warranted.
Supported by the University of Auckland Research Committeee.
TP 025
TP 026
IMPAIRMENT OF AIRWAY MACROPHAGE PHAGOCYTOSIS OF

APOPTOTIC EOSINOPHILS IN PERSISTENT ASTHMA
INTRAVENOUS IgG AMELIORATES ALLERGIC AIRWAY

INFLAMMATION VIA FCg RECEPTOR IIB
CHRISTINE VAN DALEN1, ELIZABETH HARDING1, PRACHEE GOKHALE1,

CHANI TROMOP Van DALEN1, NEIL PEARCE1, MARK HAMPTON2
1
Centre for Pubic Health Research, Massey University, Wellington 6021,
New Zealand, and 2Free Radical Research Group, Christchurch School of
Medicine, Christchurch, New Zealand
M YAMAMOTO1, K KOBAYASHI1, Y ISHIKAWA1, K NAKATA1, E

KURAMOTO1, Y KONO1, A SAKASHITA1, Y FUNADA1, Y KOTANI1,
M YOSHIDA2, Y NISHIMURA1
1
Division of Respiratory Medicine, and 2Division of Gastroenterology,
Department of Internal Medicine, Kobe University Graduate School of
Medicine, Kobe, Japan
There is evidence to suggest that impairment in macrophage phagocytosis of

apoptotic inflammatory cells is a factor in the development of chronic inflammation in asthma. We have compared pulmonary macrophage ability to phagocytose apoptotic eosinophils in non-asthmatics and intermittent and persistent
asthmatics.
Methods Cross-sectional study of 37 young adults (current asthma 21; never
asthma 16) randomly selected from a university-based general practice database. Study participants completed an asthma questionnaire, provided a blood
sample from which apoptotic eosinophils were derived, and underwent spirometry with reversibility testing followed by sputum induction with hypertonic
(4.5%) saline. Pulmonary macrophages were purified from sputum samples
and incubated with apoptotic eosinophils for one hour. Percent macrophage
phagocytosis of apoptotic cells was determined from a cytospin.
Results Median percent phagocytosis was higher in non-asthmatics compared with asthmatics (non-asthma 18.3%, IQR 11.6%; asthma 11.8%, IQR
13.8) and higher in intermittent asthmatics compared with persistent asthmatics (intermittent 11.8%, IQR 7.0%; persistent 9.3%, IQR 18.8%) although
neither difference reached statistical significance.
Conclusions These results suggest that impairment in macrophage phagocyctosis may play a role in the pathogenesis of chronic inflammation in asthma.
Supported by a grant from the Marsden Fund.
A31
Intravenous gamma globulin therapy (IVIg), which is therapeutic in a variety of

immune diseases, has been reported to be effective on patients with severe
steroid-dependent asthma. Although FceR are known to play important roles in
asthma, there are few reports about the role of Fcg?receptors in asthma. Fcg
receptor IIB (FcgRIIB) is unique inhibitory receptor, which suppresses immune
response. In this study, we evaluated the effect of IVIg in allergic airway
inflammation in OVA-challenged mice and the mechanism of the inhibitory
effects of IVIg and FcgRIIB.
Method C57BL/6 mice (WT) and FcgRIIB deficient mice (KO) were sensitized
with ovalbumin (OVA) and alum and subsequently challenged with nebulized
OVA. Before OVA challenge rabbit IgG was administered intravenously. The
airway inflammation and effects of IgG were assessed by histology, cell counts
of BAL fluid and airway hyperresponsiveness.
Result Histology showed that IgG treatment ameliorated the inflammation
around the airway and the vessels and hypertrophy of goblet cells induced by
OVA challenge. The counts of total cells and eosinophils in the BAL fluid were
reduced in IgG treated mice (total cells: 18.76 9.64 vs. 53.24 15.46,
eosinophils: 10.76 9.26 vs. 44.66 14.92 10/mL) when compared to
control mice. The reductions of cells by IgG treatment were dose-dependent
manner. However, the effect of IgG was not significant in KO mice. Total cells
and eosinophils in the BAL fluid were not reduced by IgG treatment in KO mice
challenged by OVA (total cells: 39.79 34.85 vs. 35.39 23.81, eosinophils:
33.88 32.83 vs. 24.64 18.11 10/mL).
Conclusion These data show that intravenous IgG administration suppresses the airway inflammation via FcgRIIB.
Asthma & Allergy SIG: Poster Session 2.

Genetics and Early Life and Epidemiological Studies
TP 027
GSTM1 DEFICIENCY AND TOBACCO SMOKE EXPOSURE

INCREASES SEVERITY OF CHILDHOOD ASTHMA
JASMINKA MURDZOSKA, SIEW-KIM KHOO, GUICHENG ZHANG,
JOELENE BIZZINTINO, ANDREW MARTIN, INGRID A LAING, JACK
GOLDBLATT, PETER N Le SOUF, SUNALENE G DEVADASON,
CATHERINE M HAYDEN
School of Paediatrics and Child Health, University of Western, WA 6008
Introduction Asthma is a disease influenced by multiple environmental and
genetic factors. Exposure to environmental tobacco smoke (ETS) can contribute to the development of asthma in predisposed individuals. Glutathione
S-transferase enzymes (GSTs) are important in detoxification of ETS constituents. Deletion of the GSTM1 gene results in the absence of enzyme and thus
loss of activity. The GSTM1 null genotype has been associated with asthma;
however, this has not been previously examined in children with acute asthma.
Aim To investigate if GSTM1 null influences asthma severity in acute asthmatic children who are exposed to household ETS.
Methodology Children (n = 219), aged 216 years, were recruited on presentation with acute asthma and the severity of their attack assessed. Exposure to household ETS was determined via questionnaire data, and individuals
were genotyped for the presence of GSTM1 using PCR.
Results Children with GSTM1 null (mean severity score 9.83) did not experience more severe asthma than those with at least one copy of the gene
(mean severity score 9.52, p = 0.39). The proportion of children with moderate/
severe asthma symptoms was not influenced by ETS exposure (non-ETS:
80.8%, ETS: 83.8%, p = 0.63). However, children with GSTM1 null and
xposure to ETS had a 4.69-fold (95% CI 1.1 to 19.5) increased risk of experiencing moderate/severe symptoms, compared to children with GSTM1 who
were not exposed to ETS (p = 0.033).
Conclusions These data suggest that a combination of ETS exposure and
deficiency in GSTM1 is associated with increased severity of acute asthma in
children. One possible explanation for this might be reduced efficiency of ETS
detoxification through the glutathione pathway.
TP 028
THE MIGRATORY ABILITY OF MATURE DENDRITIC CELLS IS

SUPRESSED FOLLOWING LYS-DES[ARG9]-BK STIMULATION
ROSALIND GULLIVER, SVETLANA BALTIC, KANTI BHOOLA,
PHILIP THOMPSON, MIRJANA FOGEL-PETROVIC
Lung Institute of Western Australia, Centre for Asthma, Allergy & Respiratory
Research, University of Western Australia, Nedlands, WA 6009
The migratory activity of DCs is modulated in inflammatory diseases such as
asthma. Recently, we reported that immature DCs express kinin receptors and
that bradykinin (BK) significantly enhances the migration of immature DC in
vitro. As kinins mediate many of the pathophysiological effects associated with
asthma, we hypothesized that lys-des[Arg9]-BK, which is produced during
inflammation and acts via the B1 receptor (B1R), would inhibit migration of
mature DCs.
Methods Day 7 cultured human monocyte-derived DCs were matured with
LPS, TNFa +IL-1b or CD40L in the absence or presence of lys-des[Arg9]-BK.
Maturation of DC was analysed by flow cytometry (FACS). B1R expression was
assessed by reverse-transcriptase PCR and quantitative confocal microscopy.
Migration of mature DC was assessed in transwell chambers with Lysdes[Arg9]-BK and the chemokine CCL19 used as chemoattractants.
Results Maturation of DCs was found to result in down-regulation of B1R
expression to varying degrees depending upon the maturation stimulus used.
Mature DCs all demonstrated an ability to migrate toward Lys-des[Arg9]-BK and
CCL19. However pre-treatment with Lys-des[Arg9]-BK decreased the migratory
ability of all mature DCs to both chemoattractants.
Conclusions Along with chemokines, lys-des[Arg9]-BK is likely to play a
crucial role in regulating the in vivo migration of mature DC during inflammation. The production of lys-des[Arg9]-BK during inflammation potentially immobilizes mature DCs thereby facilitating locally-mediated immune responses
within inflamed tissues.
Supported by the Asthma Foundation of Western Australia.
A32
TP 029
ROLE OF THROMBOMODULIN EXPRESSION ON DENDRITIC

CELLS IN THE PATHOGENESIS OF ATOPY AND ASTHMA
STEPHANIE YERKOVICH1,2, MARJUT ROPONEN1, MIRANDA SMITH1,
PETER SLY1, KATHY MCKENNA1, ANTHONY BOSCO1, LILY SUBRATA1,
PETER LE SOUEF1, PATRICK HOLT1, JOHN UPHAM1,2
1
Division of Cell Biology, Telethon Institute for Child Health Research, Perth
6008, and 2School of Medicine, University of Queensland, Brisbane 4102
Introduction Dendritic cells (DC) are thought to be important in allergic
diseases such as asthma, though little is known about features that distinguish
DC from atopic and non-atopic individuals, nor the mechanisms by which DC
induce or reinforce a Th2 response. We reasoned that a microarray screen of
allergen-exposed DC would highlight novel allergy-associated genes.
Methods Monocyte-derived DC were exposed to Dermatophagoides pteronyssinus allergen or LPS, and gene expression was analysed by microarray,
RT-PCR and flow cytometry.
Results Microarray analysis showed that allergen, but not LPS, enhanced
thrombomodulin (CD141) expression by DC from atopic (n = 8), but not healthy
individuals (n = 9), and was confirmed at both the mRNA (p = 0.037) and
protein level (p = 0.030). FACS sorted CD141+ DC induced a strongly Th2polarized cytokine response by allergen specific T-cells in vitro, compared to
DC lacking CD141 (n = 8). The in vivo relevance of these findings was confirmed by showing that more CD141+ DC were present in the blood of those
with stable atopic asthma (n = 13), compared to healthy subjects (n = 15), and
also that CD141 mRNA expression increased in blood leukocytes from children
during an acute asthma exacerbation and waned at follow up 6 weeks later
(n = 25).
Conclusions These findings suggest for the first time that CD141 expression
on DC may be involved in the pathogenesis of atopy and asthma.
Supported by the NHMRC; Nomination APSR Early Career Development
Award.
TP 030
GENETIC ASSOCIATIONS BETWEEN TWO PROSTAGLANDIN

RECEPTOR GENES (DP AND CRTH2) WITH ASTHMA AND
DISEASE SEVERITY
BERNADETTE BRADLEY1,2, HARLEY BLAKEMAN1,2, CAROLYN JONES3,
DAVID DUFFY4, SALLY-ANNE MCCAPPIN1, JING SHI1,2, PHILIP J THOMPSON1
1
CRC for Asthma, the Lung Institute of Western Australia, and CAARR at the
University of Western Australia (UWA), WA 6009, 2UWA School of Medicine
and Pharmacology at Sir Charles Gairdner Hospital, WAIMR and CMR at
UWA, WA 6009, 3School of Biological Sciences and Biotechnology, Murdoch
University, WA 6150, and 4Genetic Epidemiology Group, Queensland
Institute of Medical Research, QLD 4000
The inflammatory mediator prostaglandin D2 (PGD2), acting through its two
receptors PTGDR (prostaglandin D receptor) and CRTH2 (chemoattractant
receptor expressed on Th2 cells), effects asthma through chemotactic recruitment of inflammatory cells to the lung. Recently, several polymorphisms in the
two PGD2 receptor genes have been variably associated with asthma in seven
different ethnic populations.
Methods We genotyped six single nucleotide polymorphisms (SNPs) within
the two PGD2 receptor genes for their associations with asthma, disease
severity and atopy within a large (n = 1087), well-phenotyped population of
Australian Caucasian asthmatics and non-asthmatic controls.
Results One SNP (G1651A) in the CRTH2 gene was significantly associated
with both asthma (p = 0.0130) and asthma severity (p = 0.0412). None of the
other SNPs (CRTH2 G1544C, PTGDR T-549C, C-441T, T-197C or C-2T) were
associated with disease either singly or in haplotype combinations (p>0.1770).
Conclusion One SNP in the PGD2 receptor gene CRTH2 is associated with
asthma in an Australian population. This association has also been reported in
African American and Chinese populations. However, we showed no associations between asthma and SNPs in the second PGD2 receptor gene PTGDR.
Supported by the CRC for Asthma.
TP 032
TP 031
USE OF EXON EXPRESSION ARRAY TO DETERMINE THE ROLE

OF ALTERNATIVE SPLICING IN ASTHMA PATHOGENESIS
ESTRI ARTHANINGTYAS1,2, CAROLYN WILLIAMS1,2, PHILIP J THOMPSON2
Cooperative Research Centre for Asthma and Airways, and 2Lung Institute
of Western Australia, Centre for Asthma, Allergy and Respiratory Research,
University of Western Australia, Perth, Western Australia
Introduction Alternative or aberrant splicing is a major contributor to protein

diversity, in which a single gene can generate structurally and functionally
distinct protein isoforms. The role of alternative splicing in asthma pathogenesis has not been previously investigated. We hypothesized that specific alternatively spliced asthma candidate genes contribute to the development of
asthma. We chose to use a new and innovative approach involving the use of
the GeneChip(r) Exon array system together with real-time quantitative PCR to
study asthma candidate genes in human monocyte-derived dendritic cells.
Materials and Methods Asthmatic and non-asthmatic subjects provided
20 mL of blood from which peripheral blood mononuclear cells (PBMC) were
isolated by Ficoll-Paque gradient centrifugation. Monocytes were separated
from other leukocytes by adherence method, and differentiated into dendritic
cells following incubation with defined concentrations of GM-CSF and IL-4.
RNA was isolated and reverse transcribed for real-time semi-quantitative PCR
and densitometry. Chi Squared test was used to assess associations between
alternative splicing and asthma.
Results Data indicate splice variant expression in dendritic cells from asthmatic patients is influenced by asthma severity.
Conclusion Exon expression array analysis has generated a number of
asthma candidate genes with alternative splice variants. Further studies to
validate these data in a replicate data set and establish the functional significance of our findings in asthma are underway.
ABERRANT GENE ISOFORM EXPRESSION IN ASTHMATIC AND

NON-ASTHMATIC CELLS
CAROLYN WILLIAMS1,2 ESTRI ARTHANINGTYAS1,2, PHILIP J THOMPSON1,2
Cooperative Research Centre for Asthma and Airways (CRCAA), and
2
Lung Institute of Western Australia (LIWA), Centre for Asthma, Allergy and
Respiratory Research, University of Western Australia, Perth, Western
Australia
Alternative or aberrant splicing occurs in more than 70% of genes and is a

major contributor to protein diversity, in which a single gene can generate
structurally and functionally distinct protein isoforms1. The role of alternative
splicing in asthma pathogenesis has not been previously investigated. We
hypothesized that specific alternatively spliced asthma candidate genes contribute to the development of asthma. We chose to study one asthma candidate
gene in human stimulated and unstimulated: (1) monocytes, (2) monocytederived dendritic cells and (3) lung smooth muscle cells.
Methods Asthmatic and non-asthmatic subjects provided 40 mL of blood
from which peripheral blood mononuclear cells (PBMC) were isolated by FicollPaque gradient centrifugation. Monocytes were separated from other leukocytes by adherence method. Up to 50% of the monocytes were then
differentiated into dendritic cells following incubation with defined concentrations of GM-CSF and IL-4. Induction experiments used 1 mg/ml LPS and cells
were stimulated for an optimal period of 24 hrs. RNA was isolated and reverse
transcribed for real-time semi-quantitative PCR and densitometry. Chi Squared
test was used to assess associations between alternative splicing and asthma.
Results Data from stimulation experiments indicate splice variant production
can be regulated by the inflammatory response and that this response is
influenced by asthma status.
Conclusion Preliminary experiments have confirmed the presence of an
aberrant splice variant for an asthma candidate gene in the primary cells
studied. Further studies to confirm these data and establish the functional
significance of our findings in asthma are underway.
TP 033
A33
TP 034
ACUTE ASTHMATIC CHILDREN WITH CD14-260TT HAVE

LOWER IgE LEVELS AND LUNG FUNCTION
THE ROLE OF GSTP1 AND LEVEL OF ETS EXPOSURE IN

CHILDREN WITH ACUTE ASTHMA
IA LAING1,2, OEM SAVENJIE2, GL HALL2, JA BIZZINTINO2, H CLIFFORD2,

CM HAYDEN2, GC GEELHOED2, J GOLDBLATT2, FD MARTINEZ3,
PN LE SOUF2
1
Pop Sci/Genetics & Health, Telethon Inst for Child Health Res, WA 6008,
2
School of Paeds & Child Health, UWA, WA 6008, and 3Arizona Resp
Center, AZ, USA
EN NG, SG DEVADASON, SK KHOO, G ZHANG, JA BIZZINTINO,

IA LAING, J GOLDBLATT, PN LE SOUEF, CM HAYDEN
School of Paediatrics and Child Health, University of Western Australia,
Perth, Western Australia, Australia
CD14 -260CC has been associated with lower CD14 levels and higher atopy
severity in asthmatics when well. We showed in children with acute asthma,
CD14 -260CC had lower CD14 levels and higher attack severity. We hypothesized that CD14 -260CC would have increased atopy and decreased lung
function (LFTs).
Methods Children (216 yrs) were recruited on presentation to ED with acute
asthma. Rhinovirus (RV) was detected using PCR of nasal samples. Plasma
total and HDM sIgE levels (n = 165) and LFTs (n = 44, well enough to complete) were obtained within 24 hours. LFTs were measured in children, 35 yrs
with FOT (or those unable to complete FEV1) and >5 yrs with spirometry. LFTs
were quality controlled using ATS guidelines. Z-scores were calculated for FOT
resistance at 8 Hz, using local healthy children. Z-scores for FEV1 used
NHANNES 2008. Either FOT or FEV1 was used for each child.
Results Children were 30.8% female, 85.7% atopic & 79.9% had RV. Children with CD14 -260TT had a mean total IgE and HDM sIgE of 5.18 & 1.53 ku/l
compared to 5.65 & 2.53 ku/l for those with -260CT or CC (p = 0.017,
p = 0.013), respectively. Overall, children with CD14 -260TT had no difference
in LFT Z-scores compared to -260CT or CC children. However, in children with
rhinovirus detected, those with the CD14 -260TT had significantly lower mean
LFT Z-scores (Z = -3.65) compared to children with -260CT or CC (Z = -1.92,
p = 0.012). Results were consistent for both LFT methods.
Discussion The atopy results are consistent with previous studies. Therefore, despite limited numbers, it is intriguing -260TT children have lower LFTs
when infected with RV, yet well enough to complete LFTs.
Conclusion CD14 C-260T may play a role in airway function during an
asthma attack with RV.
Supported by Australian Respir. Council Ann Woolcock Fellowship & NHMRC.
Exposure to environmental factors, such as environmental tobacco smoke

(ETS), plays a significant role in modulating pre-existing genetic susceptibilities
to diseases including asthma. The Glutathione S-transferase enzymes (GSTs)
play an important role in the detoxification of ETS. There are several GST
isoforms and GSTP1 codes for the GST Pi isoform, which is the primary GST
isoform expressed in human lung tissue. Two single nucleotide polymorphisms
(SNPs) at positions 105 and 114 have been reported in GSTP1 and associated
with asthma and atopy. The aim of this study was to examine the effect of these
SNPs in combination with ETS, on asthma phenotypes in a cohort of asthmatic
children. Children were recruited during an acute episode requiring presentation at an emergency department. Genotyping using PCR-RFLP was completed on 218 children and ETS exposure was determined by parental
questionnaire. Urinary cotinine was measured in the children and was in
agreement with questionnaire responses. Statistical analyses were performed
using SPSS. There were no significant associations between the genotypes
and asthma severity during acute exacerbations. Significant associations were
found between the SNPs and atopy in this population with an odds ratio of 2.77
for the 105AA genotype (p = 0.029) and OR of 5.47 for the 114CC genotype
(p = 0.002). However, when an interaction with ETS was included, the odds
ratios increased to 9.02 for 105AA (p = 0.05) and 9.17 for 114CC (p = 0.020).
These results suggest that there is a significant gene/environment interaction
impacting on atopy in this cohort.
Funded by National Health and Medical Research Council of Australia
TP 036
ARE CHILDHOOD INFECTIONS LINKED TO ALLERGIC

DISEASES IN CHILDREN WITH A FAMILY HISTORY OF
ALLERGIES?
TP 035
A FUNCTIONAL RAGE PROMOTER POLYMORPHISM IS

ASSOCIATED WITH CHRONIC INFLAMMATORY ASTHMA
PHENOTYPES
BP SHELTON, K BALAKRISHNAN, MC HOF, C WILLIAMS, PJ THOMPSON
Lung Institute of Western Australia Inc, Perth, Australia
The RAGE gene encodes the receptor for advanced glycation end-products
(RAGE), a member of the immunoglobulin superfamily. RAGE activation by
ligands, including amphoterin and S100/calgranulins, leads to prolonged
NF-kB signalling and has been associated with chronic inflammation. Despite
high levels of RAGE expression in lung tissue, little research has been undertaken into the role of RAGE in the chronic inflammatory asthma phenotypes of
severe and aspirin-sensitive asthma.
Objective Determine genetic associations between functional polymorphisms in the RAGE promoter and severe and aspirin-sensitive asthma
phenotypes.
Methods PCR and restriction fragment length polymorphism (RFLP) were
used to genotype three RAGE promoter polymorphisms, -429T>C, -374T>A
and a 63 bp deletion from -407 to -345, in a large case-control asthma
population phenotyped for asthma severity, atopy and aspirin sensitivity.
Results No associations were identified between any of the polymorphisms
and the occurrence of asthma. However, the -374A allele was linked with both
severe asthma (P = 0.013) and aspirin-sensitive asthma (P < 0.001). Likewise,
genotypes containing the -374A allele were strongly associated with both
severe asthma (OR 2.10, 95% CI 1.323.36) and aspirin-sensitive asthma (OR
3.13, 95% CI 1.456.77).
Conclusions The -374A allele of the RAGE gene, previously shown to lead
to a 3-fold increase in promoter activity, is associated with the chronic inflammatory asthma phenotypes of severe and aspirin-sensitive asthma. These
results suggest that increased RAGE expression, with a concomitant increase
in NF-kB signalling, may in part contribute to the inflammatory response seen
in these conditions.
JA THOMSON1, C WIDJAJA1, A LOWE1, MC MATHESON1, C BENNETT1,

M ABRAMSON3, C HOSKING2, D HILL2, SC DHARMAGE1
1
MEGA Epidemiology, The University of Melbourne, Vic 3010, 2Department
of Allergy & MCRI, Royal Childrens Hospital, Parkville, Vic 3052, and
3
Department of Epidemiology and Preventive Medicine, Monash University,
Melbourne, Vic 3004
Background The global prevalence of allergic diseases is rising and
Australia has one of the highest prevalence rates in the world. The role
of early childhood infections in the development of allergic disease remains
controversial.
Objective To examine the association between early childhood infections
and the development of allergic diseases in later childhood, in high risk
children.
Methods Data were analysed from the Melbourne Atopic Cohort Study
(MACS) of 620 infants with 1 or more first-degree family members with atopic
disease. Primary risk factors assessed were otitis media, bronchitis and gastroenteritis reported in the first two years of life. Outcomes were current
asthma, hay fever and eczema at 6 years of age. Logistic regression was used
to estimate crude and adjusted odds ratios.
Results Asthma was the most common allergic condition (25.4%, 95% CI
21.629.5%), followed by eczema (24.9%, 95% CI 21.129.0%) and hayfever
(15.6%, 95% CI 12.519.1%). The most commonly reported infection was otitis
media (58.9%, 95% CI 54.962.8%), then gastroenteritis (37.7%, 95% CI
33.941.7%) and then bronchitis (19.4%, 95% CI 16.322.7%). All 3 types of
infection within the first 2 years of life were associated with increased risk of
asthma. An increased risk of asthma at 6 years was seen with otitis media
(OR = 1.14, 95% CI 1.021.3), bronchitis (OR = 1.34, 95% CI 1.01.8) and
gastroenteritis (OR = 1.23, 95% CI 0.961.6). When the frequency of infection
was examined, those who reported at least 3 episodes of gastroenteritis had a
34-fold increased risk and an almost 30% absolute increased risk (RD 0.34,
95% CI 0.080.59).
Conclusion These findings appear to contradict the hygiene hypothesis. The
findings for gastroenteritis are novel. Further examination of these associations
and possible underlying mechanisms is warranted.
Grant Support Asthma Foundation of Victoria, Nestle.
A34
TP 037
TP 038
ESTIMATION OF ASTHMA INCIDENCE IN AUSTRALIA USING

PHARMACEUTICAL BENEFITS SCHEME (PBS) DATA
PATTERNS OF ASTHMA MEDICATION USE: AN AUSTRALIAN

POPULATION-BASED LONGITUDINAL COHORT STUDY
ROSARIO D AMPON1, TERESA TO2, LEANNE POULOS1,

ANNE-MARIE WATERS1, GUY B MARKS1
1
Australian Centre for Asthma Monitoring (ACAM), Woolcock Institute of
Medical Research, NSW 2050, and 2The Research Institute, Hospital for
Sick Children, Toronto, Canada
ROSARIO AMPON1, GUY MARKS1, TERESA TO2, LEANNE POULOS1,

ANNE-MARIE WATERS1
1
Australian Centre for Asthma Monitoring (ACAM), Sydney, Australia, and
2
Hospital for Sick Children, Toronto, Canada
Background Knowledge about incident cases of asthma in Australia is

limited because they are not routinely reported. The ability to predict the
number of new cases of asthma would be helpful in allocating resources for
asthma education, management and care. Data on first use of medications for
asthma gives an indication of the incidence of asthma. The objective of this
study was to estimate the incidence rate of asthma by investigating asthma
medication use in individuals.
Methods Pharmaceutical Benefits Scheme (PBS) records for all prescriptions filled for inhaled corticosteroids (alone or combined formulation),
cromones and leukotriene receptor antagonists from July 2002 to June 2005
were included. Using a 2-year look back window, any persons who had their
first prescription for any of these drugs dispensed between July 2004 and June
2005 were assumed to be incident cases. Overall and age-specific incidence
rates were calculated per 100 asthma-medication-free individuals.
Results There were 352,082 individuals who had their first asthma medication dispensed between July 2004 and June 2005, which equates to an overall
incidence rate for asthma of 1.89 per 100. The incidence was higher among
children aged 014 years (2.07) and adults aged 65 years and over (2.45).
Conclusions Our estimated incidence rates were consistent with those
reported by others in the literature. While the PBS database was designed for
administrative purposes, it can be used to estimate incidence rates for asthma.
Support ACAM is a collaborating unit of the Australian Institute of Health and
Welfare and is funded by the Department of Health and Ageing (DoHA). We
acknowledge the Pharmaceutical Pricing and Estimates Section of DoHA for
provision of PBS data.
Keywords Asthma incidence, Pharmaceutical Benefits Scheme.
Background The ability to assess individual patterns of asthma medication

use would have clinical relevance in targeting effective asthma education and
management for this common condition. To describe longitudinal patterns of
asthma medication use, we used a population-based prescription database to
follow individuals from the first time they filled an asthma prescription.
Methods Pharmaceutical Benefits Scheme (PBS) records for all prescriptions filled for inhaled corticosteroids (alone or combined formulation),
cromones and leukotriene receptor antagonists between July 2002May 2007
were included. The cohort of incident cases comprised all individuals who filled
a prescription for one of these drugs for the first time between July 2004 and
June 2005 (determined using a 2-year (July 2002June 2004) look back
window). This cohort was followed prospectively for 2 years (to May 2007) to
examine longitudinal patterns of asthma medication use defined as one-time
use (no use after June 2005), sporadic use (had medication in only 1 of 2
follow-up years) or regular use (had medication in both follow-up years).
Results There were 352,082 individuals in the incident cohort. In the 2-year
follow-up, 61% of them were one-time users, 22% had sporadic use and 16%
had regular use. The prevalence of one-time and sporadic use was highest in
1534 year olds and regular use was most common in adults aged 65 years
and over.
Conclusions The Pharmaceutical Benefits Scheme data are valuable in
assessing population-based patterns of asthma medication use. Future
follow-up of this cohort could allow the development of statistical probabilities
to predict one-time or sporadic versus regular use of asthma medications.
Support ACAM is a collaborating unit of the AIHW and is funded by the
Department of Health and Ageing.
Keywords Asthma, Pharmaceutical Benefits Scheme.
TP 039
ASTHMA AS AN ASSOCIATED CAUSE OF DEATH IN DEATHS

ATTRIBUTED TO OTHER CAUSES
WEI XUAN, LEANNE M POULOS, ANNE-MARIE WATERS, GUY B MARKS
Medical Research, NSW 2050
Asthma is more commonly listed on death certificates as an associated cause
of death, in people whose deaths are attributed to other causes, than as an
underlying cause of death. Understanding the importance of these associations would contribute towards an overall appreciation of the impact of asthma
on mortality. The objective of this analysis was to estimate the prevalence of
asthma as an associated cause of death when various other diseases were
attributed as the underlying cause of death.
Method Multiple cause of death data were obtained from the National Mortality Database (AIHW) for the years 2001 to 2005 for all deaths in which
asthma (J45, J46) was listed as an associated cause of death. Underlying
causes for these deaths were classified by disease chapter (excluding asthma,
for the respiratory chapter).
Results Asthma was an associated cause in 4652 deaths during this period
(61.7% female, 80.7% aged 65 years and over). During this time there were
1,768 deaths attributed to asthma as an underlying cause. The underlying
causes for which asthma was most likely to be an associated cause of death
were diseases of the blood (36 cases of associated asthma, representing 1.6%
of deaths attributed to this cause), endocrine, nutritional and metabolic diseases (298 cases, 1.3%) and diseases of the circulatory system (2254 cases,
0.9%).
Conclusion Asthma is relatively uncommonly listed as an associated cause
of death in people dying of other causes. The observed differences may reflect
the true association between asthma and other diseases but possible confounding factors, such as age, need to be considered.
Keywords Mortality, Asthma as associated cause of death.
TP 040
ASSOCIATED DISEASES CONTRIBUTING TO DEATH DUE TO

ASTHMA
WEI XUAN, LEANNE M POULOS, ANNE-MARIE WATERS, GUY B MARKS
Background People whose deaths are attributed to asthma often have other
diseases that have contributed to their demise. A better appreciation of the
nature of these associations would help to frame a more holistic approach to
the management of asthma. The objective of this analysis was to estimate the
prevalence of various diseases as contributing causes of death in people
whose deaths are attributed to asthma.
Method Multiple cause of death data were obtained from the National Mortality Database (AIHW) for the years 2001 to 2005 for all deaths in which
asthma (J45, J46) was the underlying cause of death. Other listed causes were
classified by disease chapter or, within the respiratory chapter, as influenza,
pneumonia and other acute lower respiratory infection (J09-J22), COPD and
bronchiectasis (J40-J44, J47), upper respiratory tract illness (J00-J06, J30-39)
and other respiratory disease (J60-J99). The prevalence of associated causes
was estimated.
Results Asthma was the underlying cause of 1764 deaths during this period
(62.8% female, 63.7% aged 65 years). Among these there was no associated
cause of death listed in 17% of cases. Influenza or pneumonia was an associated cause in 26.1%, COPD in 9.3% and other respiratory disease in 20.8%.
The most common other system diseases that were associated were diseases
of the circulatory system (49.0%), endocrine diseases (11.8%) and mental and
behavioural diseases (12.3%).
Conclusion Care of patients with asthma should include plans to manage
associated heart disease, diabetes and psychiatric illness.
Keywords Asthma mortality, Associated cause of deaths.
TP 041
A35
TP 042
ARE HEALTH INDICATORS ON ASTHMA PREVALENCE,

MORBIDITY AND MORTALITY HIGHLY CORRELATED?
ASTHMA AND RESPIRATORY SYMPTOMS IN AUSTRALIAN

SCHOOL CHILDREN
WEI XUAN1, TERESA TO2, LEANNE M POULOS1, ANNE-MARIE WATERS1,

GUY B MARKS1
1
Australian Centre for Asthma Monitoring (ACAM), Sydney, Australia, and
2
Research Institute, Hospital for Sick Children, Toronto, Canada
ADRIANA CORTS1, PAOLA ESPINEL1, KATE HARDAKER1, BRETT TOELLE1,

GUY MARKS1, GAIL WILLIAMS2, BIN JALALUDIN3
1
Woolcock Institute of Medical Research, Sydney, NSW 2050, 2School of
Population Health, University of Queensland, Herston, QLD 4006, and
3
Centre for Research, Evidence Management and Surveillance, NSW 2170
Background ACAM currently recommend 24 indicators to measure

population-level asthma health and outcomes. We examined correlations
among several asthma indicators covering prevalence, morbidity and mortality
to try and produce a condensed set of indicators which minimized redundancy.
Methods Seven of the 24 indicators were included in this study: prevalence
of ever having doctor diagnosed asthma, prevalence of current asthma,
asthma-related general practice (GP) encounters, proportion of people with
asthma with an asthma action plan (AAP), hospitalizations for asthma, hospital
patient days for asthma, and deaths due to asthma. A correlation matrix was
created for these indicators by age groups. Pearson correlation coefficients
0.7 or -0.7 were considered strong.
Results There were strong positive correlations between prevalence of ever
asthma and current asthma (r = 1.0); GP visits and AAP possession (r = 0.74),
hospitalization (r = 0.91) and patient days (r = 0.95); and hospitalization and
patient days (r = 0.90) and AAP possession (r = 0.73).
Conclusions Only 6/21 correlations were strong, suggesting evidence for
minor redundancy among these indicators. Prevalence of ever asthma and
current asthma; hospitalization and patient days correlated perfectly and
yielded similar correlations with other indicators. Thus the inclusion of both
prevalence and both hospitalization indicators may not be necessary. Future
analyses will extend this work to include indicators that measure asthmarelated symptoms and quality of life.
Support ACAM is a collaborating unit of the Australian Institute of Health and
Welfare and is funded by the Department of Health and Ageing.
Keywords Asthma, health indicators, prevalence, mortality, morbidity.
Recent Australian reports have shown that the prevalence of asthma and
respiratory symptoms has decreased over the last 1015 years. As part of a
larger study investigating child health and air quality we have collected nationwide data from schoolchildren living in ACT, Victoria, Queensland, WA and SA.
Methods Schools were selected based on proximity to air quality monitoring
stations. Classes from years 3 to 6 were randomly selected and all children
were invited to participate. Parents self completed a questionnaire that
included questions about diagnosed asthma and respiratory symptoms.
Results A total of 1989 children provided questionnaires for analysis. The
response rate varied between states and territories and ranged from 30% to
42%. The sample comprised 51.9% girls and the mean age of children was
10.2 years.
Outcomes
Ever diagnosed asthma
Current asthma (Does he/she still have asthma?)
Wheeze in the past 12 months
Respiratory symptoms limiting activities
Missed school due to asthma or wheezing
Prevalence (%)
27.9
13.8
16.1
11.8
8.8
Conclusions Despite the relatively low participation rate, the prevalence

estimates for current asthma are similar to those reported in the National
Health Survey 200405 [1]. There is no evidence of any recent increase in the
prevalence of childhood asthma.
Funding Australian Research Council, National Environment Protection
Council.
Conflict of Interest Nil.
Reference
ACAM. Asthma in Australia: findings from the 20042005 National Health
Survey. Canberra: AIHW; 2007. (ACM 10).
TP 043
TRACKING ASTHMA SYMPTOMS AND LUNG FUNCTION FROM

AGE 7 TO 44 YRS: TASMANIAN LONGITUDINAL HEALTH
STUDY
TP 044
CHARACTERIZATION OF ASTHMA CONTROL AND AIRWAY

INFLAMMATION DURING EXACERBATIONS IN PREGNANCY
SC Dharmage1, MC Matheson1, JB Burgess1, D Mesaros2, S Morrison3,

I Feather2, R Wood-Baker2, DP Johns2, MJ Abramson4, EH Walters1,2,4
1
MEGA Epidemiology, The University of Melbourne, 2Royal Brisbane
Hospital & Gold Coast Hospital, Queensland, 3Menzies Research Institute,
University of Tasmania, and 4Epidemiology & Preventive Medicine, Monash
University, Melbourne
VANESSA E MURPHY1, VICKI L CLIFTON2, PETER G GIBSON1

Centre for Asthma and Respiratory Diseases, and 2Mothers and Babies
Research Centre, University of Newcastle, NSW 2308
Introduction Tracking the development and prognosis of a heterogeneous

disease such as asthma is important in clarifying pathogenesis of disease. The
aim of this study was to track the clinical manifestations of asthma and lung
function of the participants of the Tasmanian Longitudinal Health Study (TAHS).
Methods TAHS is a longitudinal population-based respiratory study of 8583
subjects which commenced in 1968 when they were 7 years of age. Since the
initial study another 4 follow-ups have been conducted, including the most
recent follow-up when subjects were 44 years of age. Lung function of the total
sample was measured at baseline and in sub-samples in 3 subsequent followups. Asthma was categorized as persistent, frequent or episodic when participants reported asthma symptoms in at least 3 follow-ups, in 2 follow-ups or in
1 follow-up respectively.
Results By age 7 years ever asthma prevalence was 16%. At age 44, 10%
of those who had not reported asthma by age 7 had asthma symptoms while
75% of those who reported asthma by age 7 had no asthma symptoms. Hence
over all only 25% of the asthma symptoms at age 44 were attributable to
asthma developed by age 7. In contrast, 91% of the persistent and frequent
asthmatics had developed their asthma by age 7. Persistent and frequent
asthmatics had more symptoms and poorer lung function at age 7, 14 and 44
as well as more reversibility at age 44 (p < 0.05). Childhood asthmatics who
also had a productive cough by age 7 were more likely to have persistent
asthma than those without a cough (p < 0.05).
Conclusions Although the majority of middle-age asthma is related to postchildhood onset asthma, most severe middle-age asthma has its origin in
persistent childhood disease. Having productive cough in childhood may identify high risk asthmatics who require especially rigorous management in early
life.
Grant Support NHMRC, Clifford Craig Foundation, Victorian, Tasmanian &
Queensland Asthma Foundations.
One third of women experience an improvement in asthma during pregnancy,

and symptoms improve in most women in the late third trimester. We hypothesized that the exacerbation rate would be reduced and that symptoms during
exacerbations would be less severe in the third trimester compared to the
second trimester.
Methods Pregnant women with asthma (n = 81) were prospectively followed
from recruitment (14.8 weeks (3 SD)) to delivery at clinic visits (18, 30,
36 weeks and during exacerbation), and fortnightly phone calls. The asthma
control questionnaire (ACQ) was administered at each contact and exacerbations classified as severe (requiring medical intervention) or mild (selfmanaged). Lung function, medication use, fractional exhaled nitric oxide
(FENO) and full blood counts were assessed.
Results A total of 98 mild exacerbations in 58 women (44 second trimester,
51 third trimester), and 60 severe exacerbations in 37 women (31 second
trimester, 23 third trimester) occurred. Total exacerbations, ACQ score, steroid
use and % predicted FEV1 at exacerbation were not significantly different
between trimesters. FENO and blood eosinophils at exacerbation were lower in
the third trimester compared to the second (FENO: second trimester 22.7
(median), 11.944.0 (interquartile range), third trimester 12.3, 8.018.7; Eosinophils: second trimester 0.2, 0.10.3, third trimester 0.1, 0.10.2, P < 0.05).
FENO was inversely correlated with gestational age at exacerbation (Spearman rank correlation, r = -0.38, P = 0.004).
Conclusions Contrary to our hypothesis, the total number and severity of
exacerbations was not improved in the third trimester compared to the second.
However, the exacerbations were less likely to be eosinophilic in the third
trimester.
Supported by Asthma NSW, University of Newcastle, Hunter Medical Research
Institute, Port Waratah Coal Services.
Nomination APSR Early Career Development Award.
Conflict of Interest NO.
A36
TP 045
DOES FREQUENT PARACETAMOL EXPOSURE IN FIRST TWO

YEARS OF LIFE INCREASE RISK OF CHILDHOOD ASTHMA?
ADRIAN LOWE1, JOHN CARLIN1, CATHERINE BENNETT1, CLIFFORD
HOSKING2, MICHAEL ABRAMSON3, DAVID HILL4, SHYAMALI DHARMAGE1
1
MEGA Epidemiology, University of Melbourne, 2Pediatrics, John Hunter
Childrens Hospital, 3Epidemiology & Preventive Medicine, Monash
University, and 4MCRI, Royal Childrens Hospital
Paracetamol is commonly used in infants as an analgesic and antipyretic. It
has been hypothesized that frequent paracetamol consumption may result in
reduced lung capacity to cope with oxidative stress and increase risk of
respiratory disease. To date, no study has examined exposure to paracetamol
during infancy, when lungs are still developing, and risk of childhood asthma.
Method A birth cohort of 620 infants with an atopic family history was
recruited. Frequency of paracetamol exposure was prospectively documented
up to 2 years of age. Interviews were conducted at 6 and 7 years to ascertain
asthma in the previous 12 months.
Results Paracetamol exposure in infancy was common (97% exposed by
two years of age), with some infants receiving paracetamol on up to 77 days.
The prevalence of current asthma in childhood was approximately 30% (148/
495). Frequent use of paracetamol (greater than 25 days in first two years) was
associated with increased risk of childhood asthma (OR = 1.85, 95%CI = 1.18
2.89) compared to infants who had received less than 15 days of paracetamol.
Adjustment for sex, parental history of asthma and presence of older siblings
did not greatly alter this association, but additional adjustment for use of
antibiotics and number of infections substantially reduced the strength of these
associations (OR = 1.30, 95%CI = 0.782.19).
Conclusion Frequent use of paracetamol in infancy may be associated with
increased risk of childhood asthma, and unnecessary use should be avoided.
Further work is needed to separate the effect of paracetamol exposure from the
effect of concomitant and infections.
Support Dairy Australia, CRC for Asthma and Airways, VicHealth, Nestle.
TP 046
THE INCREASED RISK OF CHILDHOOD ASTHMA ASSOCIATED

WITH FREQUENT INFANT CHILDHOOD ANTIBIOTIC EXPOSURE
IS CONFOUNDED BY EARLY LIFE WHEEZE
ADRIAN LOWE1, JOHN CARLIN1, CATHERINE BENNETT1,
CLIFFORD HOSKING2, MICHAEL ABRAMSON3, DAVID HILL4,
SHYAMALI DHARMAGE1
1
MEGA Epidemiology, University of Melbourne, 2Pediatrics, John Hunter
Childrens Hospital, 3Epidemiology & Preventive Medicine, Monash
University, and 4MCRI, Royal Childrens Hospital
It has been hypothesized that mucosal immune response requires a particular
micro-flora milieu in the infants gastro-intestinal tract, and that early life antibiotic exposure may disrupt this process and increase risk of allergic disease.
Method A birth cohort of 620 infants with an atopic family history was
recruited. Exposure to oral antibiotics was prospectively documented up to 12
months of age. Interviews were conducted at 6 and 7 years to ascertain asthma
in the previous 12 months.
Results By one year of age, approximately 80% of infants had received at
least one course of oral antibiotics. The prevalence of current asthma in
childhood was approximately 30% (148/495). Frequent use of antibiotics (more
than 20 days exposure during first year of life) was associated with increased
risk of childhood asthma (OR = 2.52, 95% CI = 1.404.54) when compared to
infant who had not been exposed. Excluding infants with a diagnosis of asthma
within the first two years of life, reduced this association by about 30%
(OR = 1.80, 95% CI = 0.903.57) and adjustment for gender, parental history of
asthma and number of infections in the first year of life further reduced this
association (OR = 1.60, 95% CI = 0.793.22).
Conclusion The increased risk of childhood asthma associated with antibiotic exposure in the first year of life is, at least in part, due to confounding with
early life wheeze and infections. If real, the independent effect of antibiotic
exposure on risk of childhood asthma is likely to be minimal in this high risk
cohort.
Support Dairy Australia, CRC for Asthma and Airways, VicHealth, Nestle.
TP 048
TP 047
EARLY AGE AT PREGNANCY IS ASSOCIATED WITH AN

INCREASED RISK OF ADULT ONSET ASTHMA
MC MATHESON1, CL WHARTON1, MJ ABRAMSON3, EH WALTERS2,
SC DHARMAGE1
1
MEGA Epidemiology, University of Melbourne, 2Menzies Research Institute,
Tasmania, and 3Epidemiology & Preventive Medicine, Monash University
Introduction The epidemiological data on asthma suggest a gender difference that varies with age. Hormonal effects have been suggested as a possible
explanation for these differences. The aim of this study was to examine reproductive factors and risk of asthma among the females of the Tasmanian
Longitudinal Health Study (TAHS).
Methods The TAHS is a longitudinal population-based cohort study of respiratory disease which commenced in 1968 when subjects were 7 years of age.
Four follow-up studies have been conducted including the current most comprehensive follow-up with subjects at 44 years of age. Information has now
been collected on reproductive factors such as number of pregnancies, age at
pregnancies, age at menarche and contraceptive pill use as well as on asthma
status. Reproductive factors were examined as risk factors for asthma using
multiple logistic regression to adjust for all likely confounders.
Results A total of 2,776 women completed the most recent postal survey. Of
these 355 (12.8%) had current asthma, and of these women with current
asthma 73.5% (261) developed asthma after childhood. On average these
women were in their mid-twenties when they developed asthma (mean SD
age = 26.6 12.5 yrs). We found with increasing age at first birth an approximate ~30% reduced risk of current asthma in women who developed their
asthma post-childhood (trend p = 0.04). We did not observe any other associations between reproductive factors and risk of asthma.
Conclusions Our results are consistent with the hypothesis that early pregnancy may promote asthma development by altering the immune response
favouring a TH2 pathway. A delay in the age of first pregnancy reduces this risk
of asthma.
Grant Support NHMRC, Clifford Craig Foundation, Victorian & Tasmanian
Asthma Foundations.
EARLY LIFE EXPOSURE TO CAT AND DEVELOPMENT OF

SENSITIZATION TO CAT
S RODRIGUES1, SC DHARMAGE1, A LOWE1, C HOSKING2, D HILL2,
MC MATHESON1
1
MEGA Epidemiology, University of Melbourne, and 2Department of Allergy,
Royal Childrens Hospital, Melbourne
Introduction The association between exposure to pets in early life and
subsequent development of sensitization and allergic disease remains controversial. The objective of this analysis was to examine the relationship between
cat exposure before birth and development of cat sensitization over time within
the Melbourne Atopic Cohort Study (MACS).
Methods The MACS is a prospective longitudinal cohort study that initially
recruited 620 women antenatal in Melbourne from February 1990 to November
1994. Detailed information on cat exposure was collected at recruitment and
frequently until two years of age. Skin prick test (SPT) were conducted at 6, 12,
24 months and 10 years. The data were analysed by logistic regression and
using Generalized Estimating Equations (GEE) for the repeated measures
design.
Results Among 620 subjects, 169 (28.8%) had a cat before birth. At 6
months, 1.9% (n = 11) of subjects were sensitized to cat and by 10 years of age
18.8% (n = 68) were sensitized. Those who did not have cat before birth belong
to a higher social class, and were more likely to have a father with allergic
disease than those with a cat. Those who developed sensitization to cat were
more likely to have a paternal family history of allergic disease and more likely
to be sensitized to other allergens. We did not observe any association
between exposure to cat before birth and the development of sensitization to
cat at 6 months (OR = 0.7, 95% CI 0.13.3), 12 months (OR = 1.4, 0.53.9), 24
months (OR = 0.76, 0.22.5) or 10 years (OR = 0.6, 0.21.4). These crosssectional results were confirmed by the GEE analysis.
Conclusion Our results fail to show an association between cat exposure
before birth and development of sensitization to cat. Furthermore exposure
after birth in the first 18months of life was not associated with an increased or
decrease risk of sensitization to cat. Our results do not support either a benefit
or risk associated with cat ownership and sensitization.
TP 049
PREMATURITY, NOT BIRTH WEIGHT, IS ASSOCIATED AN

INCREASED RISK OF ASTHMA FROM CHILDHOOD TO
ADULTHOOD
A37
TP 050
EARLY LIFE EXPOSURES AND ASTHMA REMISSION BY

MIDDLE AGE: A LONGITUDINAL STUDY OVER 37 YEARS
A LPEZ-POLN DOLHABERRIAGUE1, JA BURGESS1, CL WHARTON1,

G GILES4, JL HOPPER1, MJ ABRAMSON3, EH WALTERS2,
SC DHARMAGE1, MC MATHESON1
1
MEGA Epidemiology, The University of Melbourne, 2 Menzies Research
Institute, University of Tasmania, 3Epidemiology & Preventive Medicine,
Monash University, and 4The Cancer Council Victoria, Australia
Introduction Peri-natal events influence the development of asthma and
atopic diseases in childhood but the current literature is contradictory on the
effect of low birth weight, small for gestational age and prematurity on asthma
risk. The aim of this study was to assess the relationship between these three
exposures and asthma from childhood to adulthood.
Methods Information on asthma status of all children born in 1961 and
attending school in Tasmania in 1968 was extracted from the Tasmania Longitudinal Health Study (TAHS) (n = 8583). We obtained all available birth
records of children born in 1961 in Tasmania from the Tasmanian State
Archives and directly from Tasmanian hospitals. The total number of TAHS
participants with available birth weight was 2456 (28.6 % of the cohort) and of
these 51.2% (n = 1257) had information on gestational age. Multivariate logistic
regression was used to examine the association between prematurity and
asthma. Cox regression was used to estimate the association between prematurity and incident asthma.
Results Out of the 2456 participants in the present study, 4.7% met the
criteria for low birth weight, 8.0% were small for gestational age and 6.0% were
premature. Prematurity (<37 weeks gestation) was associated with an
increased risk of current asthma at age 7 (OR: 2.05; 95% CI: 1.024.12).
Prematurity was also associated with an approximate 2.18 (95% CI: 1.193.99)
fold increased risk of incident asthma in adult life.
Conclusions Prematurity, independent of small for gestational age, was
associated with a greater risk of asthma in early life and a greater risk of
incident asthma in adulthood. Our findings suggest that prematurity may have
an unfavourable effect on lung growth. This reduced lung growth may not
manifest as respiratory disease until later in life.
JA BURGESS1, EH WALTERS1,2,3, GB BYRNES1, MC MATHESON1,

KS ADAMS1, DP JOHNS2, MJ ABRAMSON3, SC DHARMAGE1
1
MEGA Epidemiology, School of Population Health, The University of
Melbourne, 2Menzies Research Institute, Tasmania, and 3Department. of
Epidemiology & Preventive Medicine, Monash University
Background The course of asthma may be influenced by inherited and early
life environmental factors.
Objective To examine early life exposures and asthma remission over time.
Method The Tasmanian Longitudinal Health Study (TAHS) began in 1968
when 99% (n = 8,583) of 7-year old Tasmanian school children were enrolled.
Data collected included asthma status, age at asthma onset, history of allergic
disorders, pneumonia and infant feeding and family history of asthma and
allergies. The cohort was re-surveyed in 2004 when 82% (n = 5,729) of the
7010 traced survivors completed a respiratory questionnaire that included age
at asthma remission. Time at risk of asthma remission was calculated. Asthma
remission rate ratios (RR) were estimated dependent on early life exposures.
Hazard ratios (HR) estimated asthma remission over time.
Results Asthma ever by age 44 years was reported by 27.2% (n = 1,561).
Of these, 35.6% (n = 561) had remitted. Maternal asthma (RR 0.59; 95% CI
0.440.78), maternal atopy (0.66; 0.470.91), maternal smoking (0.79; 0.63
0.99), childhood allergic rhinitis (0.47; 0.350.62), infantile eczema (0.73; 0.55
0.93), flexural eczema (0.72; 0.550.93) and food allergy (0.68; 0.480.98) all
predicted a lesser likelihood of remission. Independent predictors of a lesser
likelihood of remission were maternal asthma (HR 0.67; 95% CI 0.490.91),
childhood allergic rhinitis (0.57; 0.420.78) and maternal smoking (0.79; 0.63
1.00).
Conclusion Genetic predisposition, childhood allergic rhinitis and exposure
to maternal smoking were associated with a lesser likelihood of asthma remission by middle age. Aggressive management of childhood allergies and discouragement of maternal smoking may increase asthma remission over time.
Grant Support NHMRC, Clifford Craig Foundation, Royal Hobart Hospital
Research Foundation, Victorian & Tasmanian Asthma Foundations.
TP 051
THE PREVALENCE OF ASTHMA IN ADULTS AND CHILDREN

THE BUSSELTON 20052007 SURVEY
AL JAMES, M KNUIMAN, M HUNTER, J HUI, DE WHITTALL, M DIVITINI,
E INGLIS, AW MUSK
Busselton Population Medical Research Foundation, Departments of
Pulmonary Physiology and Respiratory Medicine, Sir Charles Gairdner
Hospital, School of Public Health, University of Western Australia, Nedlands,
WA 6009
Background The increasing global prevalence of asthma over the last
23 decades may have slowed in developed countries recently. Respiratory
symptoms, doctor-diagnosed asthma (DDA), atopy and airway hyperresponsiveness (AHR) have been assessed in the Busselton Health Studies
since 1966.
Aim To assess the current prevalence of DDA, wheeze (<12 months), atopy
and AHR in children and adults in Busselton.
Methods An age- and sex-stratified random sample of adults, selected from
the electoral roll, was invited to complete a questionnaire and attend the local
study centre for assessment of atopy (allergen skin tests) and AHR (methacholine). All children from participating primary and secondary schools were
also invited to attend. The prevalences of DDA, wheeze, atopy, AHR and
current asthma (wheeze + AHR) were calculated.
Results Adults prevalences were: DDA17% males (M), 20% females (F),
decreasing until age 40 years; wheeze25% M, 22% F; AHR12% M, 14%
F; current asthma4% M, 4% F; atopy52% M, 44% F. For adults 1855
years old, DDA was 22.0%. Primary school children: DDA27% M, 19% F;
current wheeze18% M, 12% F; AHR20% M, 18% F: current asthma3%
M, 4% F, atopy49% M, 35% F. Secondary school children: DDA27% M,
22% F; wheeze17% M, 14% F; AHR19% M, 18% F; current asthma 4% M,
7% F; atopy51% M, 44% F. Approximately a third of adults and children who
had current asthma did not report DDA.
Conclusions Asthma prevalence remains high in adults and children and for
adults 1855 years has continued to increase from 9% in 1981 to 16% in 1990
to 22% in 2006 in the Busselton community. A significant proportion of cases of
current asthma may not be receiving or seeking appropriate medical attention.
TP 052
RISK FACTORS FOR ADULT ONSET ASTHMA: A 14-YEAR

LONGITUDINAL STUDY IN THE BUSSELTON POPULATION
EUZEBIUSZ JAMROZIK1, MATTHEW W KNUIMAN1,2, ALAN JAMES1,4,
MARK Divitini2, AW (Bill) MUSK1,2,3
1
Busselton Health Study, Sir Charles Gairdner Hospital, Nedlands, WA,
2
School of Population Health, University of Western Australia, Nedlands,
WA, 3Department of Respiratory Medicine, Sir Charles Gairdner Hospital,
Nedlands, WA, and 4West Australian Sleep Disorders Research Institute,
Nedlands, WA
Background Few longitudinal studies have examined the risk factors and
natural history of adult-onset asthma. This retrospective case-control
population-based study of a large cohort of adults examined lifestyle factors
and subject characteristics associated with adult-onset asthma.
Methods Longitudinal data on 1,554 adults from the Busselton Health Study
surveys in 1981 and 1994/95 who did not report ever having had asthma in
1981 including questionnaire measures of doctor-diagnosed asthma, allergic
rhinitis, respiratory symptoms, snoring and smoking. Lung function was
assessed by spirometry and atopy by skin-prick tests. Logistic regression was
used to identify baseline risk factors as well as changes that were associated
with the onset of asthma.
Results Wheeze, allergic rhinitis, chronic cough, smoking and reduced lung
function in 1981 predicted asthma diagnosis by 1994/95. The onset of asthma
was associated with new wheeze, new rhinitis, weight gain, new snoring, and
greater decline in lung function. Neither pre-existent nor new onset atopy were
associated with the onset of asthma.
Discussion The results of this study add to existing knowledge concerning
the links between upper and lower respiratory tract disorders, and demonstrate
that atopy as measured by skin-prick tests is not associated with adult-onset
asthma.
Conclusion These findings suggest that different factors give rise to the
onset of asthma in adults than those previously observed from studies in
children. Upper airway disorders such as rhinitis and sleep apnoea may be
more important than atopy in determining the risk of adult-onset asthma.
A38
Asthma & Allergy SIG: Poster Session 3. Physiology,

Environment, Investigation and Management
TP 053
TP 054
NON-RESPIRATORY COMORBIDITY IN ADULTS HOSPITALISED

WITH ASTHMA IN AUSTRALIA [267]
EXPIRATION TO RV ABOLISHES DEEP INSPIRATION

BRONCHOPROTECTION IN NON-ASTHMATICS
ANNE-MARIE WATERS, LEANNE M POULOS, WEI XUAN, GUY B MARKS

DAVID CHAPMAN1,2, NORBERT BEREND1, GREG KING1,

BRENT MCPARLAND2, CHERYL SALOME1,2
1
Woolcock Institute of Medical Research, Sydney, Australia and
2
Department of Pharmacology, University of Sydney, Australia
Background Asthma is often associated with comorbidity, however few

studies have investigated comorbidities among people with this common condition. The objective of this analysis was to describe patterns of non-respiratory
comorbidity among adults hospitalized with asthma in Australia.
Methods Data on hospitalizations for people aged 15 years and over with a
principal diagnosis of asthma (J45, J46) were obtained from the Australian
Institute of Health and Welfares (AIHW) National Hospital Morbidity Database
for the period 200506. Patterns of comorbidity were examined by investigating
additional diagnoses for non-respiratory disease according to ICD-10 diseasespecific chapters.
Results Among people aged 15 years and over hospitalized in 200506 with
a principal diagnosis of asthma (16,566 hospitalizations; 70% female; 47%
aged 3564 years), 33% had at least one non-respiratory comorbidity. Median
length of stay was higher among those with at least one comorbidity (4 days)
than among those with no comorbidities (2 days). Among people aged 1564
years, the most common comorbid condition was endocrine, nutritional and
metabolic diseases (19%), while among those aged 65 years and over it was
diseases of the circulatory system (32%).
Conclusions A large proportion of asthma hospitalizations in Australia are
associated with non-respiratory comorbidity and a longer length of stay.
Further, the pattern of non-respiratory comorbidity associated with asthma
hospitalizations varies by age. Given our rapidly ageing population, the level of
comorbidity associated with asthma has implications for coordinated health
care and demand on health services.
Keywords Comorbidity, Hospitalization, Asthma.
Background Deep inspirations to TLC (DI) reduce airway responsiveness to

methacholine (Mch) in non-asthmatic subjects. We hypothesize that expiration
to RV, rather than FRC, following DI may modify any protective effect of DI.
Aim To compare the effects of DI followed by expiration to FRC (DI-FRC) and
to RV (DI-RV) on Mch-induced airway closure and airway narrowing in nonasthmatic subjects.
Methods 10 non-asthmatic subjects had incremental Mch challenges (dose
range 0.8 to 200 mmol) to determine PD20FEV1. The PD20 dose or, if a 20%
fall (FEV1) was not reached, a dose of 102.2 mmol, was used in subsequent
single dose challenges. After 20 minutes of DI avoidance, subjects inhaled
single dose Mch immediately (no DI) or after 5 DI-FRC. In addition, 6 nonasthmatic subjects underwent a single dose Mch challenge after 5 DI-RV.
Airway narrowing was measured by FEV1 (% fall) and airway closure by FVC
(% fall)/FEV1 (% fall). All data are reported as mean 95% CI.
Results Compared to no DI, 5DI-FRC reduced both airway narrowing
(12 5% fall vs 23 11% fall, p = 0.04 ) and airway closure (0.24 0.10 vs
0.58 0.11, p < 0.0001). The protective effects of DI were abolished when DIs
were followed by expiration to RV, so there was no significant difference
between 5DI-RV and no DI either for airway narrowing (29 12% fall vs
23 11%, p = 0.95) or airway closure (0.54 0.19 vs 0.58 0.11, p = 0.16).
Conclusion In non-asthmatic subjects the protective effect of deep inspiration on airway narrowing and airway closure is abolished if deep inspiration is
followed by a full expiration to RV. We speculate that expiration to RV introduces ventilation heterogeneity and/or airway closure, which may occur
through the deactivation of surfactant compounds.
TP 056
AIRWAY WALL AREA MEASURED BY HRCT INCREASES AFTER

BRONCHODILATOR AND LUNG INFLATION IN ASTHMATICS
TP 055
DO VIRAL INFECTIONS INFLUENCE THE SEVERITY OF ACUTE

ASTHMA EXACERBATIONS IN NON-HOSPITALISED CHILDREN?
AB CHANG1, R CLARK2, JP ACWORTH2, H PETSKY1, TP SLOOTS3
Depts of Respiratory Med, 2Emergency Medicine, 3Qld Paediatric Infectious
Diseases Laboratory, Royal Childrens Hospital, Brisbane, Queensland
Background Asthma exacerbations are often triggered by viral respiratory

infections, yet the influence of respiratory infections on the morbidity of acute
asthma beyond the immediate period is unknown. We examined the influence
of nasopharyngeal (NPA) respiratory viral, Chlamydia and Mycoplasma detection on asthma morbidity in children presenting to the Emergency Department
for an acute exacerbation of asthma.
Methods A subset (n = 78) of the 201 children enrolled for a randomized
controlled trial (RCT) on the efficacy of 5 vs 3 days of oral prednisolone had an
NPA taken at presentation. NPA were examined for Chlamydia, Mycoplasma
and respiratory viruses (enteroviruses, coronaviruses, human metapneumovirus, adenovirus, parainfluenza, influenza, RSV, rhinoviruses) by PCR. Enrolled
children were aged 216 years with recurrent wheeze and required 600 ?g
(MDI/spacer) or 2.5 mg (nebulized) of salbutamol to reduce tachypnoea.
Parents filled validated diary cards for cough and asthma severity, and completed asthma QOL data at enrolment and end of weeks 1 and 2.
Results PCR for various viruses was positive in 42 (53.8%) children, with no
significant difference in the groups the children were randomized into. Rhinovirus PCR was positive in the NPA of 32 children, RSV in 7, hMPV in 2,
adenovirus, parainfluenza, influenza A and B in one each. Specimens were
negative for the other micro-organisms listed above. Children with a NPA viral
positive state were significantly (p = 0.002) younger than those with a negative
state. However, there was no difference in the any of the asthma outcomes of
children whose NPA was positive or negative for the micro-organisms tested.
Conclusions In children with an acute asthma exacerbation presenting to
emergency health facilities, a respiratory virus could be identified in >50% but
the presence of a respiratory virus did not influence the morbidity of the asthma
exacerbation at presentation or at the end of week-1 and week-2.
NATHAN J BROWN1,2, CHERYL M SALOME1,2, ANEAL CHANDRA1,2,

NORBERT BEREND1,2, GREGORY G KING1,2,3
1
Woolcock Institute of Medical Research, Camperdown NSW 2050,
2
The University of Sydney, NSW 2006, and 3Royal North Shore Hospital,
St Leonards, NSW 2065
Airway wall thickness measured using HRCT is reported to be increased in
asthmatic compared with control subjects. However, it is unknown whether wall
thickness is a fixed structural characteristic of the airways or if it responds to
transient changes in bronchomotor tone or airway size.
Aim To determine the effects of bronchomotor tone and lung volume on
airway wall area measured by HRCT.
Methods 8 patients with doctor-diagnosed asthma had partial chest HRCT
scans, before and after bronchodilator (BD), at FRC, TLC and a volume
midway between (MID-volume). Airway segments were identified between
branch points and matched between consecutive lung volumes both before
and after BD, and also at constant lung volume before and after BD. Mean
lumen areas and wall areas for each airway segment at each volume were
measured using automated analysis software. Paired t-tests were used to
determine changes due to BD and lung inflation.
Results 44 airways were matched before and after BD at FRC. Absolute
airway wall area (WA) was related to airway lumen diameter (Di). Proportional
airway wall area (WA%) was inversely related to Di. BD increased Di at FRC
from (Mean SD) 4.58 0.60 mm to 5.23 0.63 mm, p < 0.0001, WA from
31.15 4.33 mm2 to 34.71 4.33 mm2, p < 0.0001, and decreased WA%
from 64.8 2.7% to 61.0 2.6%, p < 0.0001. Lung inflation increased Di
before and after BD (p < 0.0001). Lung inflation from FRC to MID-volume
increase WA after BD (p = 0.003) but not before. Lung inflation from MIDvolume to TLC increased WA before BD (p = 0.013) but not after.
Conclusion In asthmatics, airway wall area increases in response to BD and
to increasing lung volume. Measures of wall thickness by HRCT may be
confounded by bronchomotor tone or lung volume.
Supported by The CRC for Asthma and Airways and The University of Sydney
Bridging Grant U1089.
Nomination None.
Conflict of Interest: None.
A39
TP 057
TP 058
ASSOCIATIONS BETWEEN WOOD SMOKE AIR POLLUTION AND

RESPIRATORY SYMPTOM PREVALENCE IN TASMANIA
FIXED AIRWAY OBSTRUCTION ASTHMA AND COPD:

DIFFERENCES IN FUNCTIONAL EXERCISE CAPACITY AND
LUNG FUNCTION
CM BENNETT1, S DHARMAGE2, MC MATHESON2, J GRAS3,

D MESAROS4, C WHARTON2, J MARKOS4, J HOPPER2, EH WALTERS1,2,4,
MJ ABRAMSON1
1
Department of Epidemiology & Preventive Medicine, Monash University,
VIC 3004, 2Centre for Molecular, Environmental, Genetic and Analytic
Epidemiology, University of Melbourne, VIC 3010, 3CSIRO Marine &
Atmospheric Research, VIC 3195, and 4Menzies Research Institute,
University of Tasmania, TAS 7000
S Turner1,2, K Hill1,3,4, P Eastwood1,4,5, A Cook1,2,6, P Thompson1,2,

S Jenkins1,2,3
1
School of Physiotherapy, Curtin University, Bentley, WA 6845, 2Lung
Institute of WA, Nedlands, WA 6009, Depts. of 3Physiotherapy and
4
Pulmonary Physiology, Sir Charles Gairdner Hospital, Nedlands, WA 6009,
and Schools of 5Anatomy and Human Biology and 6Population Health,
University of WA, Crawley, WA 6009
Wood smoke air pollution is of concern with respect to respiratory health due to
its complex chemical composition and potential to carry air toxics into the lower
respiratory system. Launceston has a long history of poor winter air quality,
primarily due to use of domestic wood heaters. Participants in Hobart had a
similar prevalence of wood heater use, but Hobart does not experience the
same wood smoke pollution (due to differences in regional geography). The
research aim was to investigate if the prevalence of respiratory symptoms
varied between Launceston and Hobart.
Methods Data on respiratory symptoms were gathered from the 2004 postal
survey of the Tasmanian Longitudinal Health Study, which began in 1968.
Participants are now aged 46. This analysis examined 601 residents of
Launceston and 1067 residents of Hobart. Symptom prevalence and associated factors were compared between the two cities using odds ratios and c2
tests.
Results Launceston and Hobart participants were similar with respect to
atopy, smoking, existing respiratory disease and type of home heating. There
was little difference in symptom prevalence between the cities, although cough
with phlegm (OR 1.37, 95% CI 1.111.69, p = 0.004) and nocturnal shortness
of breath (OR 1.45, 95% CI 1.012.08, p = 0.05) were paradoxically significantly more prevalent in Hobart.
Conclusions Slightly higher symptom prevalence in Hobart suggests that
other factors, such as local vehicular and industrial air pollution, are more
important than wood smoke in respiratory morbidity.
Supported by NHMRC, CSIRO, Clifford Craig Foundation, Tasmanian and
Victorian Asthma Foundations.
Functional exercise capacity was assessed in patients with fixed airway

obstruction asthma (FAOA) and chronic obstructive pulmonary disease
(COPD) with a similar magnitude of air trapping. The hypothesis was despite
similarities in air trapping severity and hyperinflation, the preservation of gas
transfer in FAOA patients would result in better exercise performance.
Methods Sixteen subjects (10 male) aged 68.7 8.2 yrs with FAOA (RV/
TLC 130 19% pred) were matched by gender, age (5 yrs) and degree of air
trapping (RV/TLC, 0.05%) to 16 subjects with COPD (68.0 8.3 yrs,
RV/TLC 132 20% pred). Subjects completed two six minute walk tests
(6MWT), resting lung function tests, handgrip dynamometry and quadriceps
strength measures. HR was continuously monitored throughout the 6MWT and
dyspnoea and SaO2 were recorded pre- and post-test. *p < 0.05.
Results 6MWT
Distance Test 1 and 2 (m)
%Pred 6MWD
Peak HR (bpm)
Post SaO2 (%)
Dyspnoea Score
Asthma
COPD
541 92, 569 90

90 10*
130 20
94.7 1.9*
2.7 1.8*
466 109, 485 113

75 16*
130 14
84.3 2.7*
5.9 2.5*
Conclusions Despite a similar degree of air trapping, FAOA subjects had a

significantly greater 6MWD and less dyspnoea than subjects with COPD.
Normal SaO2 in the FAOA cohort during exercise reflects well preserved gas
transfer. Individuals with FAOA are likely to tolerate a much higher intensity of
exercise than their counterparts with COPD.
Supported by the NHMRC and Sir Charles Gairdner Hospital Research
Foundation.
TP 059
EXERCISE TRAINING IMPROVES QUALITY OF LIFE IN

MIDDLE-AGED AND OLDER ADULTS WITH MODERATE TO
SEVERE ASTHMA
S TURNER1,2,3, P EASTWOOD1,4,5, A COOK1,2,6, P THOMPSON1,2,
SS JENKINS1,2,3
1
Institute of WA, Nedlands, WA 6009, Depts. of 3Physiotherapy and
4
Pulmonary Physiology, Sir Charles Gairdner Hospital, Nedlands, WA 6009,
and Schools of 5Anatomy and Human Biology and 6Population Health,
University of WA, Crawley, WA 6009
There is limited evidence of the effectiveness of supervised exercise training in
middle-aged and older adults with moderate to severe asthma.
Methods 34 asthmatic subjects (15 male) aged 67.8 10.6 yrs, FEV1
59 16% pred, participated in a randomized controlled trial of exercise training. Assessments of quality of life (QOL, [AQLQ]), functional exercise capacity
(six minute walk distance [6MWD]), asthma control and anxiety and depression
were completed at baseline, immediately following (6 wks), and 3 mths after
the intervention period.
Results Clinically and statistically (P < 0.05) significant improvements in
QOL were observed in the exercise group at 6 wks compared to the control
group. This difference was not maintained at 3 mths. 6MWD improved at 6 wks
and 3 mths in the exercise group (p < 0.01), however the difference between
groups was not significant. In the exercise group there was a trend towards
improved asthma control and a reduction in anxiety and depression that was
not observed in the control group. *P < 0.05, change at 6 wks vs baseline;
#
P < 0.05, Exercise vs Control
Change in QOL and 6MWD at
6 wk
AQLQ Total score
AQLQ Activity domain
AQLQ Symptom domain
6MWD (m)
Exercise (n = 19)
Control (n = 15)
0.78 0.56*#
1.01 0.64*#
0.62 0.53*#
36.6 39.5*
0.27 0.4*
0.58 0.51*
0.12 0.48
10.4 36.1
Conclusions Supervised exercise training is beneficial in this population,

however the improvements in QOL, 6MWD, and asthma control decline when
supervised training ceases. A high 6MWD at baseline in this cohort may have
reduced the responsiveness of this test for detecting improvements following
training.
Supported by the NHMRC and Asthma Foundation of WA.
TP 060
MEDICATION USE BETWEEN ASYMPTOMATIC AND

SYMPTOMATIC ASTHMA PATIENTS
R KANDANE1, M MATHESON1, J SIMPSON1, M TANG2, D JOHNS3,
D MESAROS3, R WOOD-BAKER3, I FEATHER4, S MORRISON5,
M ABRAMSON6, S DHARMAGE1, E WALTERS1,2,3
1
MEGA Epidemiology, Uni of Melbourne, 2Dept Allergy & Immunology, RCH
Melbourne, 3Menzies Research Institute, Tasmania, 4Gold Coast Hospital,
5
Uni of Queensland, and 6Epidemiology & Preventive Medicine, Monash
University, Melbourne
Introduction Asthma is a chronic inflammatory disorder, which has become
a major public health problem in Australia. The objective of this analysis was to
examine asthma symptoms and lung function in relation to medication use
within the Tasmanian Longitudinal Health Study (TAHS).
Methods Spirometry (pre- & post-bronchodilator) and medication history
were collected in a sub-sample of 1281 subjects who were part of the most
recent TAHS laboratory follow-up. Subjects were categorized according to the
presence of asthma symptoms in the last year (asymptomatic/symptomatic)
and lung function (FEV1% 80 or < 80 % predicted).
Results Lung function data were available for 1116 participants. A total of 55
subjects reported symptoms and had pre-BD FEV1 < 80% predicted, 118
reported symptoms with normal lung function, and 98 had FEV1 < 80% without
symptoms. Overall 93% of symptomatic subjects with FEV1 < 80% were on
asthma medication (any) in the last 12 months (44% relievers only; 9.1%
preventers only, 38% both), compared to 75% of those with symptoms and
normal FEV1 (38% relievers only; 7% preventers only, 29% both). Of those
without symptoms but a FEV1 < 80%, 12% had used asthma medication in the
last year (7% relievers only; 1% preventers only, 3% both).
Conclusion Our results demonstrate that a lack of symptoms does not
always indicate the absence of obstructive lung function. Use of appropriate
asthma medications, preventers in particular, was found to be inadequate.
Large numbers of asymptomatic subjects with impaired lung function highlights
the possible need for spirometry screening of middle-aged adults.
Grant Support NHMRC, Clifford Craig Foundation, Victorian, Tasmanian &
Queensland Asthma Foundations.
A40
TP 061
TP 062
COMPLETENESS OF WRITTEN PEAK FLOW AND SYMPTOM

DIARIES IN AUSTRALIAN CHILDREN
AGREEMENT BETWEEN WRITTEN DIARIES AND

ELECTRONICALLY RECORDED PEAK EXPIRATORY FLOW
PAOLA ESPINEL1, ADRIANA CORTS1, KATE HARDAKER1, BRETT TOELLE1,

ADRIAN BARNETT2, GUY MARKS1, GAIL WILLIAMS2
1
Woolcock Institute of Medical Research, Sydney, NSW 2050, 2 School of
Population Health, University of Queensland, Herston, QLD 4006
KATE HARDAKER1, PAOLA ESPINEL1, ADRIANA CORTS1, HELEN REDDEL1,

BRETT TOELLE1, ADRIAN BARNETT2, GAIL WILLIAMS2, GUY MARKS1
1
Woolcock Institute of Medical Research, Sydney, NSW 2050 and 2School of
Population Health, University of Queensland, Herston, QLD 4006
Home asthma monitoring is important for measuring day-to-day variation in

lung function and symptoms. This approach requires the availability of complete diaries for a comprehensive assessment. We assessed the completeness
of written diaries collected as part of a nation wide study of air quality and child
health.
Methods Children who had ever been diagnosed with asthma and had
respiratory symptoms in the last year were identified from a cross-sectional
study. These children were asked to record symptom scores and peak expiratory flows twice daily in diaries for a five week period. The diaries and peak flow
devices were explained at a face-to-face meeting with parents and children.
Each week diaries were mailed back and parents received a phone call to
encourage completion. Completeness was defined as no missing responses to
symptom questions or peak flow measurements in diaries from week two to
week five.
Results Data from the first 36 children (822 day records) were available for
analysis. The sample included (53%) girls, mean age 10 yrs. The overall
frequencies for complete records were; morning symptoms 85%, morning peak
flow 85%, evening symptoms 83% and evening peak flow 82%. There was a
significant trend for more complete morning peak flow records over the four
weeks (Cochrane-Armitage trend test p < 0.007). Agreement between morning
and evening symptom completeness and between morning and evening peak
flow completeness was fairly poor (Kappa < 0.30).
Conclusions The completeness of symptom and peak flow records collected
in this study was very high. The comprehensive follow-up protocol implemented is likely to have had an important impact on the completeness of
asthma diaries. The availability of complete data justifies the time and cost
involved.
Supported by Australian Research Council and National Environment Protection Council of Australia.
Daily peak expiratory flow (PEF) monitoring has been used in epidemiological
studies to assess changes in lung function over time. The value of written PEF
diaries has been questioned because of problems with completeness and
validity. This study aimed to compare stored electronic PEF data and a written
diary record of those data in a panel study in children with weekly reminders to
aid adherence.
Methods Children who had ever been diagnosed with asthma and had
respiratory symptoms in the last year were identified in a population study.
They were given electronic PEF devices with a digital readout (MiniWright
Digital, MWD, Clement Clarke, UK) and written symptom and peak flow diaries
and instructed in their use at a meeting with parents and children. Each child
was asked to complete three PEF manoeuvres every morning and evening for
five weeks and to record these in the written diary. The highest PEF was
automatically stored in the MWD device with date and time. The extent to which
written and electronic records agreed was assessed. Participants were not
informed of the recording capability of the MWD.
Results A mean of 24 days (range 831) had written data available for
analysis. The sample included 53% girls, mean age 10 years. For 708 written
records of morning PEF, there was no corresponding electronic record in 177
(25%). For 692 written evening PEF records, there was no electronic record in
149 (22%). For those sessions with both written and electronic records, there
were 58 (11%) morning sessions and 79 (15%) evening sessions where the
written and electronic records differed by >10 L/min. Overall, 67% of written
entries were accurate relative to the electronic record.
Conclusions Data fabrication and data entry errors can be avoided, and thus
more accurate data obtained, if electronic PEF devices are used in place of
paper diaries.
Support Australian Research Council and National Environment Protection
Council of Australia.
TP 063
EVALUATION OF A PSYCHOLOGICAL INTERVENTION IN

ADULTS WITH ANXIETY AND ASTHMA TARGETING
ASTHMA-RELATED QUALITY OF LIFE
VANDANA DESHMUKH1, BRETT TOELLE1, TIM USHERWOOD2,
BRIAN OGRADY3, CHRISTINE JENKINS1
1
Woolcock Institute of Medical Research, Sydney, NSW 2050, 2Western
Clinical School at Westmead Hospital, University of Sydney, Sydney, NSW
2145, and 3SWAMHS Anxiety Treatment and Research Unit, Sydney, NSW
2150
Background Previous research suggests that comorbid anxiety is associated with lower asthma-related quality of life (AQOL) in adults with asthma.
However, research is scant on the role of psychological interventions in these
patients.
Aim To evaluate the effectiveness of a four-session cognitive-behavioural
therapy (CBT) intervention, in improving the AQOL, in participants with anxiety
and asthma.
Method Participants identified with comorbid anxiety and asthma were randomly assigned to the CBT intervention group (n = 10) and the asthma monitoring control group (n = 8) and evaluated on AQOL measures, at various
intervals.
Results Nine participants, in the CBT group, completed the study. Seven
participants showed a clinically significant improvement in asthma-related
emotional functioning (EF) and six participants in total AQOL scores, at the
5-week post-intervention assessment. Additionally, six participants in the CBT
group indicated clinically significant improvement in EF and five participants in
total AQOL scores, at the 3-month follow-up assessment. Only three participants in the control group completed the study. None of these participants
showed any improvement in AQOL scores at the 5-week or 3-month
assessment.
Conclusion This pilot study suggests that a higher number of participants
in the CBT group showed clinically significant improvement in EF and total
AQOL scores with higher retention rates. Further research needs to confirm
these findings in a larger group, identifying the elements of a successful CBT
intervention and characteristics of participants who respond to the CBT
intervention.
TP 064
TREATMENT OF GASTRO-OESOPHAGEAL REFLUX DOES NOT

IMPROVE ASTHMA OUTCOMES: A SYSTEMATIC REVIEW
KE LEADBEATER1, BR DAUGHERTY1, PG GIBSON2
University of Newcastle, Callaghan, NSW 2308, Australia and 2Department
of Medicine, John Hunter Hospital, Locked Bag 1, Hunter Region Mail
Centre, NSW 2310, Australia
1
Gastro-oesophageal reflux disease (GORD) is a risk factor for uncontrolled

asthma. We conducted an update of a systematic review to assess whether
treatment of gastro-oesophageal reflux in subjects with asthma improved
asthma outcomes.
Methods Randomized controlled trials (RCTs) of GORD treatment in adults
or children that reported asthma health outcomes and had symptomatic GORD
were included and assessed in accordance with the standard Cochrane systematic review process. Subjects received pharmacological therapies compared with conservative management.
Results From 261 potentially relevant studies, 19 RCTs were included in the
review. When compared to placebo, morning peak expiratory flow did not
significantly improve (change from baseline WMD 10.43, 95% CI: -9.55 to
30.42) with proton pump inhibitor treatment (n = 7 trials involving 739 participants). Asthma exacerbations were not significantly less in the intervention
groups compared with the control groups (odds ratio 0.66; 0.411.08; n = 557).
Conclusions While some trials reported evidence of asthma improvement
with GORD therapy, overall there appears to be no statistically significant
evidence of a beneficial effect. It is clear that not all persons with GORD and
asthma will gain improved control over their asthma with GORD therapy;
this may be due to the heterogeneous pathophysiology of asthma. Future
large-scale trials would be required to demonstrate an effect on asthma
exacerbations.
KEL and BRD were supported by a Cochrane Airways Group Scholarship.
TP 065
SALBUTAMOL DELIVERED VIA METERED DOSE INHALER AND

SPACER VERSUS NEBULIZER TO INDUCE
BRONCHODILATATION DURING SPIROMETRY, A CROSSOVER
STUDY
A41
TP 066
THE EFFICACY OF USING LABA/ICS COMBINATION

THERAPY AS PART OF A WRITTEN ASTHMA ACTION PLAN
HEATHER POWELL1, PETER G GIBSON1,2
Department of Respiratory and Sleep Medicine, John Hunter Hospital and
Hunter Medical Research Institute, NSW. 2University of Newcastle,
Callaghan, NSW
1
MURAD IBRAHIM, ELEONORA DEL COLLE, FRANCIS THIEN

Department of Respiratory Medicine, Box Hill Hospital, Victoria 3128
Background The ATS/ERS task force recommend the use of metered dose
inhaler (MDI) and spacer for airflow limitation reversibility testing. Salbutamol
given via MDI & spacer has been shown to be equivalent to a nebulizer in the
clinical setting. This has not been well studied in respiratory laboratory setting.
Aim To compare the 2 methods of reversibility testing in a laboratory setting.
Methods We conducted a laboratory based crossover study in a secondary
hospital. Patients with asthma or COPD were eligible. The patients firstly
underwent spirometry and reversibility testing following a standard dose of
nebulized salbutamol. They were asked to return for a second set of spirometry
within the same week and at the same time of day when reversibility with an
MDI and spacer was recorded. We used an incremental dose of salbutamol
starting from 2 puffs and up to 8 puffs. Spirometry parameters were recorded
10 minutes after each intervention. The primary outcome was the percentage
change in FEV1 after each intervention. Side effects were monitored for.
Results Nine patients with asthma were recruited. The mean percentage
change in FEV1 was higher in the nebulizer group than after only 2 puffs via
MDI & spacer (15.4 7.4 vs 6.2 8 [mean SD], p = 0.67). However, there
were no differences between the 2 arms following higher doses of bronchodilator via MDI & spacer. The mean percentage change in FEV1 after 4, 6 and 8
puffs were 12.6 11.3, 15.4 12.3, and 17.7 13.6 respectively (p = 0.09,
0.05 and 0.07 respectively when compared to the nebulizer group).
Conclusion Using an MDI and spacer for bronchodilator reversibility is
equivalent to that of a nebulizer and should be the standard method of testing.
The dose of bronchodilator needs to be at least 4 puffs as recommended by the
ATS/ERS; however 6 puffs correlated best with a standard nebulizer route.
Further increments in bronchodilator dose provided little additional bronchodilatation. The study was limited by the small number of patients.
Asthma guidelines recommend a stepwise approach to treatment. The role of

inhaled corticosteroid (ICS) and long-acting beta-agonist (LABA) combination
therapy in asthma written action plans is not clear.
Objective To assess the efficacy of adjusting ICS/LABA combination therapy
in a written action plan compared to fixed dosing in people with asthma
requiring maintenance ICS.
Methods Cochrane systematic review of randomized controlled trials comparing ICS/LABA combination therapy in a single inhaler that is adjusted up or
down according to a written action plan (WAP) to comparison 1: budesonide/
formoterol given as a fixed maintenance dose (FD) (n = 9) or comparison 2:
fluticasone/salmeterol FD (n = 2).
Results 10 parallel randomized controlled trials describing 11 interventions
met the inclusion criteria. For the trials that compared WAP to FD budesonide/
formoterol there were significant reductions for the WAP group in exacerbations, (RR (95%CI): 0.82 (0.70 to 0.97)), severe exacerbations (RR (95%CI):
0.61 (0.37 to 0.99)) and study medications (WMD (95%CI): -1.18 (-1.23
to -1.14)) with no difference in asthma control or adverse events. The results
for the two trials reporting WAP budesonide/formoterol to FD fluticasone/
salmeterol were discordant and a homogenous pooled result could not be
determined.
Conclusion Inhaled budesonide/formoterol can be effectively used as part of
a written asthma action plan.
Funding The Centre for Clinical Research Excellence.
TP 068
PREDICTORS OF POOR ASTHMA CONTROL AND URGENT

HEALTH CARE FOR ASTHMA
TP 067
IMPROVING THE QUALITY AND CONTENT OF GENERAL

PRACTICE CONSULTATIONS FOR OLDER PEOPLE
WITH ASTHMA
DIANNE GOEMAN1, ROBYN OHEHIR1, LENA SANCI2, SIMON SCHARF3,
CHRISTINE JENKINS4, JO DOUGLASS1
1
Airmed, The Alfred Hospital, Monash University, Melbourne Vic 3004, 2Dept
of General Practice, The University of Melbourne, Vic 3000, 3Aged Care
Services, Caulfield General Medical Centre, Caulfield, Vic 3162, and 4The
Woolcock Institute, The University of Sydney NSW 2050
Of the 318 Australians who died from asthma in 2005, over two thirds were over
50 years of age. This trend resulted in the National Asthma Council of Australia
(NAC) calling for better management of asthma in the elderly. We designed
an educational intervention using evidence based educational strategies to
improve the content and style of general practice consultations for older people
with asthma.
Methods Randomized controlled trial of a multi-faceted program consisting
of a group educational session, a videotaped standardized Simulated Patient
Consultation, followed by an Academic Detailing session. Forty-two GPs were
randomized into an active or a control group. GPs provided the names of
patients who would be happy to participate in the study and the program was
evaluated by patient and GP outcomes.
Results GPs recruited into our program reported improvements in a range of
clinical areas. One hundred and ten patients were recruited, their outcomes are
under analysis.
Conclusion GPs were overwhelmingly positive about participation in this trial
and our intervention successfully improved the capacity and confidence of
GPs to deliver care to older people with asthma. Our study also developed
several tools that would enable dissemination of our findings.
Supported by an Asthma Targeted Intervention Grant (ATIG) provided by the
Commonwealth Department of Health and Ageing and the Co-operative
Research Centre for Asthma and Airways.
HK REDDEL1, SZ BOSNIC-ANTICEVICH2, P CORRELL1, BG TOELLE1,

CR JENKINS1, V KRITIKOS2, RD AMPON1, M DEPHOFF3, J THOMSON3,
GB MARKS1
1
Woolcock Institute of Medical Research, NSW 2050, 2Faculty of Pharmacy,
University of Sydney, NSW 2006, Asthma Foundation of NSW, NSW 2065
In studies where direct clinical assessment is not possible, urgent health care
utilization (HCU) is often used as an indirect measure of asthma control. This
study aimed to identify factors predicting urgent HCU and asthma control.
Methods Patients in NSW with a doctor diagnosis of asthma were recruited
from community pharmacies, a research volunteer database, and databases of
Asthma Foundation NSW, to complete a questionnaire about asthma. Poor
asthma control was defined as Asthma Control Questionnaire (ACQ) score
1.5. Urgent HCU was defined as hospitalization, ED visit, or urgent doctor
visit due to asthma. Multiple logistic regression was used to identify predictors
of poor control and urgent HCU.
Results Questionnaires were completed by 608 adults (61% female) with a
doctor diagnosis of asthma (Pharmacy 260, Woolcock 299, Asthma Foundation
87). 87% used inhaled corticosteroid (ICS) long-acting b2-agonist in the last
4 wks. Median age was 56 yrs (range 1287), and 9% were current smokers.
Mean ACQ score was 1.4 (95% CI 1.31.5), with 40% of participants having
poor asthma control (ACQ 1.5). 28% had urgent HCU for asthma in the
previous year. Significant independent predictors for poor asthma control were
younger age, current smoking, living in more disadvantaged areas, being
retired, having only primary education, and holding a concession card. Predictors for urgent HCU were younger age, being in full-time employment, having
only primary education, and being of non-English speaking background.
Neither ICS use nor possession of a written asthma action plan was associated
with lower risk for either poor asthma control or HCU.
Conclusions Poor asthma control is common in NSW even in patients using
inhaled corticosteroids. Although urgent HCU is often used as an indirect
measure of poor asthma control, it is affected by different factors, perhaps
because health care utilization represents a more complex balance between
need and access.
Funded by the Asthma Foundation of NSW.
A42
TP 069
AN ABBREVIATED MANNITOL CHALLENGE USING THE

FORCED OSCILLATION TECHNIQUE
CHERYL SALOME1,2,3, MELISSA MCCLEAN1,3, CHRIS HTUN1,3,4,
NORBERT BEREND1,2,3, GREG KING1,2,3,4
1
Woolcock Institute of Medical Research, 2University of Sydney,
3
Co-operative Research Centre for Asthma and Airways, Camperdown,
NSW 2050 and 4Royal North Shore Hospital, St Leonards, NSW 2065
Bronchial challenge tests with mannitol, to measure airway hyperresponsiveness, can take up to 30 minutes and require inhalation of up to 635 mg of
mannitol. Our aim was to determine if positive mannitol challenges can be
detected after half the maximal dose (315 mg) using the forced oscillation
technique (FOT) to measure response.
Methods 15 non-asthmatic subjects and 52 asthmatic subjects underwent
standard mannitol challenge, up to 635 mg mannitol. Respiratory system conductance (Grs) and reactance (Xrs) was measured by FOT at 6 Hz during
40 sec tidal breathing immediately after each dose of mannitol. FEV1 was
measured after FOT, within 90 sec of mannitol administration. Two point dose
response slope (DRS), was calculated for Grs (DRSGrs) and Xrs (DRSXrs)
for standard tests, up to 635 mg, and for short tests by excluding data
from doses above 315 mg. Ability to detect a positive test, defined as
PD15FEV1 < 635 mg, was determined by the area under the ROC curve
(AUC) and repeatability by intra-class correlation coefficient (ICC).
Results 32 asthmatic and 2 non-asthmatic subjects had positive tests, with
PD15 FEV1 values from 9.2 to 622 mg. AUC (95%CI) did not differ between
standard (std) and short tests for DRSGrs (p = 0.14) or DRSXrs (p = 0.29)
AUC
ICC
TP 070
PATTERN OF USE OF MEDICATIONS FOR ASTHMA IN 2006
DRSGrs std
DRSGrs short
DRSXrs std
DRSXrs short
0.81, 0.680.93
0.74
0.75, 0.620.88
0.58
0.91, 0.840.99
0.86
0.87, 0.810.97
0.85
Conclusion Using DRSXrs to measure the response to mannitol challenge

may allow the use of an abbreviated challenge without reduction in repeatability
or in the ability to detect a positive response.
Supported by Pharmaxis Ltd.
Nominations None.
CHRISTINE COWIE1, NECTARIOS ROSE2, ADRIANA CORTES1, WAFAA EZZ1,

GUY MARKS1
1
Woolcock Institute of Medical Research, Sydney, NSW 2050 and 2NSW
Health Department
Combined use of inhaled steroids (ICS) and long acting beta-agonists (LABA)
have an important role in asthma management. We used data from a 2006
population sample to examine medication use in adults and children.
Methods All adults (1875 years) and children (217 years) from within four
discrete zones in northern Sydney were eligible for an interview survey, as part
of a study investigating health effects associated with traffic-related air pollution. The prevalence of use of short-acting beta-agonists (SABA), any ICS
(alone or combination) and combined formulations of ICS/LABA in the previous
three months was estimated for the study population and those with diagnosed
asthma.
Results There were 806 children [mean (SD) age 8.7 (4.6) years and 50%
female] and 2184 adults [mean (SD) age 45.6 (14.9) years and 56% female]
interviewed in 1843 households, representing an overall response rate of 33%.
The prevalence of ever diagnosed asthma was 16.1% in children and 17.4% in
adults. Medication data were missing for 301 subjects.
Table 1
Use of asthma medications in the preceding three months by age group

Adults
SABA
Any ICS
ICS/LABA
Children
% of all
(n = 1965)
% of asthma
(n = 234)
172
115
81
8.8
5.9
4.1
68.0
54.3
32.9
% of all
(n = 724)
% of asthma
(n = 77)
73
36
15
10.1
5.0
2.1
79.2
59.7
19.5
Conclusions Just over half of adults and children with diagnosed asthma
reported that they had used ICS in the preceding three months. As expected,
among adults most of these had used the combined LABA-ICS formulation
whereas among children most used ICS alone.
Funding CRC Asthma & Airways; NSW Health.
TP 071
A REVIEW OF THE RECRUITING STANDARD CONCERNING

ASTHMA FOR THE AUSTRALIAN DEFENCE FORCE (ADF)
LOUIS IRVING1, MARIKO KOH1, PETER NASVELD2, RUTH MCLAUGHLIN2,
SANDRA ANDERSON3, CHRISTINE JENKINS3, KAREN DONALD1,
JENNY FIRMAN4
1
Royal Melbourne Hospital, Parkville, 2Centre for Military and Veterans
Health, University of Queensland, 3Sydney South West Area Health Service,
and 4Defence Force Recruiting
Background Asthma affects 1:9 adult Australians and is a leading cause of
rejection for recruitment into the Australian Defence Force (ADF). Within this
diagnosis there is a wide spectrum of disease activity and clinical outcomes.
Also asthma assessment and management has improved so that many asthmatics are now fully active without any significant disruption or risk to their lives.
Hypothesis: There is a subgroup of asthmatics who are at very low risk from
significant adverse effects from asthma and who could be considered for
recruitment to the ADF.
Aims 1. To identify the subgroup of asthmatics who could be considered for
recruitment to the ADF. 2. To develop an assessment process to identify this
subgroup (screening). 3. To develop a process to evaluate the outcomes of any
change to the recruitment standard for asthma (evaluation).
Methods 1. A literature review of the natural history, assessment, management and response to treatment of mild episodic and mild persistent asthma.
2. A literature review of asthma in the military. 3. A clinical review of the outcomes of known asthmatics in the ADF. 4. An expert group to review the above
and to develop a screening process and an evaluation of the program.
Results The literature review identified a subgroup of asthmatics, defined as
mild episodic and mild persistent, who with appropriate management, have a
low risk of significant adverse asthma outcomes. They can be identified by a
combination of questionnaire, spirometry and bronchial provocation testing. A
screening process has been developed which allows asthmatics to be recruited
with a negative mannitol or hypertonic saline challenge on 400 mg/day or less
of budesonide (or equivalent) without LABA. A methodology to evaluate the
impact of these changes on the recruitment standard has also been developed.
Conclusion The recommendations were approved by the Minister for
Defence on 27/7/2007 and are now ADF policy.
TP 072
IMPACT OF ALEXITHYMIA ON SELF-MANAGEMENT AND

COMMUNICATION WITH HEALTH CARE PROVIDERS
KELLY CHUGG1, CHRISTOPHER BARTON1, RAL ANTIC2, ALAN CROCKETT1
Discipline of General Practice, University of Adelaide, and 2Thoracic
Medicine, Royal Adelaide Hospital
1
Alexithymia is a personality trait associated with difficulty identifying and communicating emotional and physical feelings. It has been associated with poor
control of asthma and near fatal asthma. The primary objectives of this study
were to: (1) identify alexithymia in a cohort of Australian asthma patients; (2)
investigate the relationship between alexithymia and asthma control; (3) investigate the relationship between alexithymia and asthma management.
Methods Cross sectional study of 25 moderate to severe asthma patients
recruited from Royal Adelaide Hospital Outpatients. Participants were either
mailed the questionnaire pack or completed it after a clinic appointment.
Existing validated questionnaires were used. Statistical analyses were performed using SPSS.
Results 11 male (44%) and 14 female (56%) patients with moderate to
severe persistent asthma (mean age 44 years, SD = 11) participated.
= 48.3, SD = 13.2). 12% (n = 3)
Alexithymia scores ranged from 23.0 to 76.0 (X
of participants could be classified high alexithymia, 32% (n = 8) borderline
alexithymia and 56% (n = 14) were low alexithymia. Alexithymia mean scores
were not statistically different across sociodemographic variables. A positive
correlation/association was found between alexithymia score and asthma
control score (r = 0.57, p < 0.01), quality of life (r = -0.65, p < 0.01), and adherence (p = 0.03) but not satisfaction with communication (r = -0.27, p = 0.2) or
number of hospitalizations (p = 0.25).
Conclusions This is the first Australian study to identify alexithymia among
asthma patients and investigate relationship to control as well as management
and communication. Associations between alexithymia and asthma control
were confirmed. A larger sample size is needed to determine impact of
alexithymia on self-management and provision of clinical care for asthma.
Grant Support KC supported by PHCRED; CB supported by Lubims
fellowship.
A43
Cell Biology/Immunology SIG: Poster Session

TP 074
TP 073
EFFECTS OF PORT HEDLAND AND URBAN DUST ON

CYTOKINE PRODUCTION BY HUMAN AIRWAY EPITHELIAL
(A549) CELLS
1
S AGGARWAL , N MISSO , J SUTTON , P THOMPSON

Lung Institute of Western Australia, Centre for Asthma, Allergy &
Respiratory Research, The University of Western Australia, Nedlands, WA
6009, 2Department of Environment and Conservation, Kensington, WA 6151
T-CELL GRANZYME B IS RELATED TO SEVERITY OF AIRFLOW

OBSTRUCTION IN COPD
J AHERN, G HODGE, M HOLMES, P REYNOLDS, S HODGE
Department of Thoracic Medicine, Royal Adelaide Hospital and Lung
Research Laboratory, Hanson Institute, Adelaide, South Australia
Port Hedland is impacted by iron-containing dust particles (PM10) that may

activate lung cells when inhaled. Furthermore, the effects of Port Hedland PM10
may differ from the effects of urban PM10 impacting metropolitan areas. The
aim of this study was to assess the effects of Port Hedland PM10 on production
and release of the inflammatory cytokines, IL-6 and IL-8, by human airway
epithelial (A549) cells, and to compare these with the effects urban PM10 from
metropolitan areas.
Methods Human airway epithelial (A549) cells were exposed to PM10
collected at Port Hedland and at urban locations (Sydney, Perth). A549 cells
were exposed to a range of PM10 concentrations (20200 mg/ml) for 24 h.
Lipopolysaccharide (LPS) and phorbol myristate acetate (PMA) were used as
positive controls. Supernatants from cell cultures were assayed for IL-6 and
IL-8 using specific ELISA kits. RNA was extracted and reverse transcribed to
cDNA. IL-6 and IL-8 mRNA expression was quantified by duplex real-time PCR
using TaqMan primer/probes.
Results LPS stimulated a 2.7-fold increase in IL-8 release and PMA stimulated a 3-fold increase in IL-8 release and a 30-fold increase in IL-6 release.
However, neither Port Hedland PM10 nor urban PM10 stimulated concentration
dependent release of IL-6 or IL-8 by A549 cells. Expression of IL-6 or IL-8
mRNA was also not altered by Port Hedland or urban dust.
Conclusions Port Hedland PM10, at concentrations up to 200 mg/ml, does
not appear to stimulate inflammatory cytokine production by airway epithelial
cells. At these levels of exposure, Port Hedland PM10 may not activate lung
epithelial cells.
Supported by the Department of Environment and Conservation, Western
Australia
CD8+ T-cells may cause airway epithelial cell apoptosis via the granzyme
pathway. We have reported increased apoptosis of airway epithelial cells and
increased BAL T-cell expression of granzyme b in COPD, and a positive
correlation between the two. We hypothesized that the increased granzyme b
would also be related to smoking history (pack years - Pk/y), age and severity
of airflow obstruction (FEV1 %pred) in patients with COPD. We further hypothesized that the T-cell granzyme b expression would be higher in the airway than
the peripheral blood.
Methods We investigated T-cell intracellular granzyme b expression in blood
from COPD subjects (33 current and 24 ex-smokers) and 12 never-smoker
controls, and bronchoalveolar lavage (BAL) and bronchial brushing (intraepithelial T-cells) from a cohort of these subjects using flow cytometry. Correlations between granzyme b and Pk/y, age or FEV1 were performed using
Spearmans rank correlation. Granzyme b in T-cells from blood, BAL and
bronchial brushings were compared.
Results There were significant correlations between FEV1 and granzyme b
expression in blood and BAL (blood: r -0.444, p = 0.002; BAL: r -0.368,
p = 0.029). There was a significant correlation between Pk/y and granzyme b
expression in blood (r 0.362, p = 0.002), but not in BAL. There were
no significant correlations between granzyme b and age. There were no significant differences in granzyme b expression in blood, BAL or intra-epithelial
compartments.
Conclusion Granzyme b is expressed at similar levels in blood, BAL and
intra-epithelial compartments, supporting recent opinion that COPD is a systemic disease. T-cell granzyme b is related to severity of airflow obstruction and
smoking history in patients with COPD and may be one mechanism of apoptosis leading to lung injury and airflow obstruction in COPD.
TP 075
TP 076
CYCLIN D1 UPREGULATION IN AIRWAY SMOOTH MUSCLE

FROM ASTHMATICS IS INSENSITIVE TO CORTICOSTEROID
INHIBITION
CORTICOSTEROIDS INHIBIT S1P-INDUCED IL-6 SECRETION IN

HUMAN AIRWAY SMOOTH MUSCLE: ROLE OF MKP-1
JC ALLEN1, T SCHLOSSER, EE RAMSAY1, Q GE2, AJ AMMIT1

Respiratory Research Group, 1Faculty of Pharmacy and 2Department of
Pharmacology, University of Sydney, NSW 2006 Australia
J PARMENTIER1, T QUANTE1, EE RAMSAY1, S HENNESS1, JC ALLEN1,

Q GE2, CL ARMOUR1, AJ AMMIT1
Respiratory Research Group, 1Faculty of Pharmacy and 2Department of
Pharmacology, University of Sydney, NSW 2006, Australia
As development of remodelled airways is correlated with deterioration of lung

function, we require therapies that reduce and reverse structural changes in
remodelled airways. In asthma, corticosteroids can halt some, but not all,
aspects of airway remodelling. Therefore, in order to aid future design of
efficacious anti-remodelling agents we need a better understanding of the
molecular mechanism/s underlying the development of airway remodelling and
the effectiveness of corticosteroids. Hyperplasia of airway smooth muscle
(ASM) is a feature of the remodelled airway in asthmatics. In this study we
examined the effect of corticosteroids on a key regulator of G1 progression
cyclin D1. ASM cells from n = 8 non-asthmatics and n = 7 asthmatics were
pretreated for 1 h with vehicle or dexamethasone (0.1 mM). The temporal
kinetics of cyclin D1 mRNA and protein expression were measured up to 24 h
after stimulation with the mitogen platelet-derived growth factor-BB (PDGFBB). PDGF-BB induced a significant increase in cyclin D1 mRNA expression in
ASM from non-asthmatics (2.6 0.3-fold) and asthmatics (2.9 0.3-fold) after
24 h stimulation. In non-asthmatics, the corticosteroid dexamethasone significantly (P < 0.05) reduced the amount of cyclin D1 mRNA expressed (to
1.6 0.2-fold). In contrast, cyclin D1 expression in asthmatics was relatively
resistant to inhibition by dexamethasone; the amount of PDGF-BB-induced
cyclin D1 expression in the absence or presence of dexamethasone was not
significantly different (2.9 0.3-fold vs. 2.2 0.3-fold), respectively. Cyclin D1
protein results support the mRNA data. These results corroborate earlier evidence demonstrating that corticosteroids inhibited proliferation only in ASM
cells from subjects without asthma (Roth, M., et al. 2004. N Engl J Med
351(6):560574) and suggest that there are corticosteroid-insensitive proliferative pathways in asthmatics that warrant further investigation.
Sphingosine 1-phosphate (S1P), a bioactive sphingolipid found elevated in the

airways of asthmatics, modulates myriad airway smooth muscle (ASM) functions that promote inflammation and remodelling in asthma. In this study, we
uncover the molecular pathway/s underlying S1P-induced secretion of IL-6,
and investigate if, and how, corticosteroids inhibit IL-6 secretion. Using cultured
ASM cells from non-asthmatics, we found that S1P induces IL-6 secretion from
ASM cells via CRE, but not AP-1, C/EBP or NF-kB, transcriptional regulation
of IL-6 gene expression. CRE-dependence was supported by S1P-induced
CREB phosphorylation. Although the corticosteroid dexamethasone reduced
S1P-induced IL-6 secretion in a dose-dependant manner, this inhibition
appeared to occur via a pathway independent of CREB/CRE, suggesting the
existence of a parallel pathway. As we recently discovered that the antiinflammatory actions of corticosteroids in ASM can be mediated via the induction of the endogenous mitogen-activated protein kinase (MAPK) inhibitor,
MAPK phosphatase-1 (MKP-1), we investigated whether MAPK represents the
parallel pathway targeted by corticosteroids. We found that S1P can induce
activation of a variety of MAPK, however, only p38 MAPK phosphorylation was
inhibited by dexamethasone; importantly, the increase in MKP-1 after corticosteroid treatment appeared to mirror the decrease in S1P-induced p38 MAPK
phosphorylation. Furthermore, exogenous expression of MKP-1 inhibited S1Pinduced IL-6 secretion. Taken together, these results suggest that parallel
pathways exist to induce IL-6 secretion (transcriptional via CREB/CRE and
possibly post-transcriptional via p38 MAPK) and serve to underscore the
importance of MKP-1 upregulation as a mechanism of action of corticocosteroids in ASM.
Supported by the NHMRC, University of Sydney Sesqui R & D Scheme.
A44
TP 077
TP 078
ESTABLISHING PRIMARY SMALL AIRWAY EPITHELIAL CELL

CULTURES FROM LUNG TRANSPLANT RECIPIENTS
TUMSTATIN, AN ANGIOGENIC INHIBITOR, MODULATES AIRWAY

ANGIOGENESIS
B BANERJEE1, A KICIC2, B RHODES1, M MUSK1, E SUTANTO2, S STICK2,

DC CHAMBERS1
1
WA Lung Transplant Program, RPH, Perth, WA and 2TICHR, Perth, WA
S BOUSTANY1,2, BG OLIVER1,2, LM MOIR1,2,3, JL BLACK1,2,3,

PM HANSBRO1,4,5, NG HANSBRO1,4,5, PS FOSTER1,4,5, JK BURGESS1,2,3
1
CRC for Asthma and Airways, 2Discipline of Pharmacology, University of
Sydney, Australia, 3Woolcock Institute of Medical Research, Australia,
4
Centre for Asthma & Respiratory Disease, University of Newcastle,
Australia, and 5John Curtin School of Medical Research, Australian National
University, Canberra, Australia
Obliterative bronchiolitis, the predominant cause of chronic allograft dysfunction, is primarily a disease of small airways and yet to date in vitro & ex vivo
primary studies utilize large airway epithelial cells (AEC). Our aim was to
establish methods for the collection & culture of primary human small airway
epithelial cells (SAEC).
Methods Bronchial brushings (3) were collected during routine bronchoscopy from subsegemental (AEC) and then (using a new brush & radiologic
guidance, 13 cm from the pleural surface) SAEC. Cells were immediately
processed & cultures established. Epithelial lineage was confirmed using
immunohistochemistry (IHC).
Results 16 patients (6 female; age 1861; 2 COPD, 6 pulmonary fibrosis, 2
cystic fibrosis, 6 other) 3118 months post transplant underwent 32 bronchoscopies. AEC and SAEC sampling was well tolerated with no complications.
Mean cell recovery for AEC (1.501 0.083 106) was higher than for SAEC
(0.822 0.072 106, p < 0.01) as was the culture establishment rate (78%
(n = 32) vs 62% (n = 16) respectively, p < 0.05). Bacterial & fungal infections
were the factors limiting successful culture passage. Cultures reached confluence after a median 31 (2068) days and maintained a typical polygonal
cobblestone pattern to passage 3. No morphological differences were seen
between AEC & SAEC & epithelial lineage was confirmed by IHC.
Conclusions Collection and culture of primary transplanted small airway
epithelial cells in humans is feasible and is well tolerated. Further studies
utilizing this technique may provide new insights into the pathogenesis of
obliterative bronchioltis.
Supported by an ADA Bartholomew Medical Research Trust Grant and The WA
Heart and Lung Transplant Foundation.
Angiogenesis is a hallmark feature of asthma. Angiogenic promoters, such as

VEGF and TGFb are reported to be increased in airways of asthmatics.
Tumstatin, an endogenous angiogenic inhibitor, is the non-collagenous
domain-1 (NC1) of the alpha3 chain of collagen IV. Decreased levels of collagen IV have been reported in the airways of asthmatics. We investigated the
presence of tumstatin in the airway of asthmatics and its potential role as an
angiogenic inhibitor.
We detected the six a chain NC1domains of Col IV and the 7S domain of the
a3 chain using immunohistochemistry. The level of tumstatin in serum and
BAL-f was measured by dot blot. Western blots were used to identify the
association with the rest of the collagen IV molecule. A tube formation assay
using primary pulmonary endothelial cells (PPEC) was performed to evaluate
the role of tumstatin in the airway. The effect of intranasal tumstatin on airway
hyperresponsiveness and angiogenesis was studied in an ovalbumin mouse
model. Tumstatin was absent in the airways of asthmatics (n = 14) while the
remaining six collagen IV a chains were present. The 7S domain of the a3
chain was present in the asthmatic airway (n = 6). Tumstatin was detected in
both serum and BAL-f samples from asthmatic volunteers (n = 10), however
the level of expression was not significantly different from that in nonasthmatics (n = 7). In asthmatic serum tumstatin was part of the whole collagen
IV a3 chain. Tumstatin was able to inhibit PPEC tube formation in a dose
related manner. Tumstatin inhibited angiogenesis in the mice airways and was
associated with an improvement in AHR. The fact that tumstatin is absent from
asthmatic airways and inhibited airway hyperresponsiveness and angiogenesis
may indicate potential for therapeutic intervention in airway remodelling.
This work was supported by the CRC for Asthma and Airways and NH&MRC.
TP 080
TP 079
RHINOVIRUS USES EGFR/HER SIGNALLING NETWORKS TO

COORDINATE IL-8 AND ICAM-1 EXPRESSION IN BRONCHIAL
EPITHELIAL CELLS
ELEVATED SAA, BUT NOT IP-10, DISCRIMINATES BETWEEN

NON-PATHOGEN AND PATHOGEN-ASSOCIATED ACUTE
EXACERBATIONS OF COPD (AECOPD)
K LIU1, RC GUALANO1, ML HIBBS3, GP ANDERSON1,2, S BOZINOVSKI1

Departments of 1Pharmacology and 2Medicine, University of Melbourne,
VIC 3010, Australia, and 3Ludwig Institute for Cancer Research, Melbourne
Branch, Parkville, VIC 3050, Australia
S BOZINOVSKI1, M THOMPSON3, A HUTCHINSON3, R VLAHOS1,

J BLACK3, D SMALLWOOD3, LB IRVING3, GP ANDERSON1,2
Depts of 1Pharmacology and 2Medicine, University of Melbourne, Victoria
3010, Australia and 3Dept. Respiratory Medicine, Royal Melbourne Hospital,
Victoria 3050, Australia
Introduction Epithelial EGFR (epidermal growth factor receptor) expression

correlates with disease severity and neutrophil infiltration in asthmatic airways.
Acute exacerbations of asthma and COPD are also associated with steroid
refractory neutrophilic inflammation, with rhinoviruses being the most common
trigger.
Methods We explored the role of EGFR signaling in regulating the key
neutrophil mediators, IL-8 and ICAM-1 in rhinovirus (RV16 serotype) infected
bronchial epithelial cells, BEAS-2B.
Results RV16 infection stimulated IL-8 and ICAM-1 expression, which was
further elevated (2-fold) by transient upregulation of EGFR levels. Detection of
viral RNA by QPCR confirmed that enhanced expression was not associated
with increased viral replication. EGFR ligands (epiregulin, amphiregulin and
HB-EGF) were induced by RV16 infection and inhibition of metalloproteases
responsible for ligand shedding partially suppressed this response. The EGFR
inhibitor AG1478, completely blocked IL-8 and ICAM-1 expression to basal
levels, as did the specific Erk1/2 inhibitor U0126. The p38 MAPK inhibitor
SB203580 blocked IL-8 secretion but not ICAM-1 expression, whereas the
PI3K inhibitor Wortmannin was ineffective in both responses. Kinase inactive
K721R EGFR which is selectively deficient in STAT signaling reversed RV16
responses associated with EGFR over-expression.
Conclusions RV16 infection rapidly promotes induction of EGFR/HER
ligands. EGFR activity is required for IL-8 and ICAM-1 induction and elevated
EGFR levels augment this response. Erk1/2 is a major EGFR effector pathway
and STAT factors coordinate gene expression associated with EGFR overexpression. These results suggest that targeting EGFR may provide a selective
therapy that dampens neutrophil-driven inflammation without compromising
essential antiviral pathways mediated by pathogen recognition receptors such
as TLR3.
Supported by NHMRC and CRC-CID.
Introduction Since AECOPD cause local and systemic inflammatory

responses, the clinical utility of predictive blood biomarkers is currently under
evaluation. We recently identified SAA as a biomarker of AECOPD but there
are currently no markers that can discriminate the involvement of pathogens
or discern between bacterial or viral causes. IP-10 (IFN-induced protein 10)
has recently been suggested as a selective biomarker of rhinovirus-induced
asthma exacerbations (Wark, JACI 07). Therefore, we evaluated serum levels
of IP-10 compared to SAA, CRP and IL-6 during AECOPD.
Methods Fifty-one blood samples from the Melbourne Longitudinal Community Cohort (MLCC) were collected from 31 individuals with Moderate-Severe
COPD during (i) an AECOPD and (ii) when clinically Stable. Respiratory virus
detection was performed by multiplex PCR on nasal swabs, and sputum culture
was used for bacterial isolation and characterization, at both time-points.
Results All blood markers were significantly elevated during AECOPD vs.
Stable (median IP10: 165.3 vs. 72.7 ng/L, CRP: 8 vs. 4 mg/L, SAA: 29.4 vs.
6.7 mg/L and IL-6: 5.8 vs. 3.6 ng/L). Since IL-6 stimulates the acute phase
response, we correlated its levels with the other markers. Only CRP was
strongly correlated with IL-6 (Spearman r = 0.58, P < 0.0001), suggesting
differential regulation of SAA and IP10. SAA discriminated between NonPathogen (n = 10) vs. Pathogen-associated (n = 41) events (SAA: 9.4 vs.
44.1 mg/L P = 0.005), whereas no significant change was observed in the other
markers (IP-10: 139.8 vs. 170.5 ng/L, CRP: 4 vs. 10 mg/L, IL-6: 4.6 vs. 7.2 ng/
L). However when AECOPD marker levels were stratified on the basis of
pathogen type (Viral = 12, Bacterial = 21, Viral and Bacterial = 8), none of the
markers were significantly altered.
Conclusions IP-10 is significantly elevated during an AECOPD, however
only SAA differentiated non-pathogen from pathogen associated events.
Supported by NHMRC and CRC-CID.
No Conflict of Interest.
TP 081
A45
TP 082
AIRWAY REMODELLING IN VITRO IS NOT ALTERED BY

COMBINED LONG ACTING 3 AGONISTS AND
CORTICOSTEROIDS
NOD-CB17prkdscid MICE LACKING FUNCTIONAL T AND B

CELLS DISPLAY COPD-LIKE INFLAMMATION AFTER SMOKE
EXPOSURE
JK BURGESS1,2, BG OLIVER1, Q GE1, S BOUSTANY2, V YOUNG1,

LM MOIR2, JL BLACK1,2
1
Discipline of Pharmacology, University of Sydney, NS, 2006, and 2Woolcock
Institute of Medical Research, Sydney, NSW 2006
JS CHAN1, R VLAHOS1, D HAYLOCK3, S NILSSON3, I BERTONCELLO3,

GP ANDERSON1,2
Departments of 1Pharmacology and 2Medicine, University of Melbourne,
Parkville Vic 3010 and 3Australian Stem Cell Centre, Monash University,
Clayton, Vic 3800
Background Severe persistent asthma is characterized by structural

changes in the airwaysairway remodelling. Airway smooth muscle (ASM)
cells have the potential to play a key role in these processes through the
release of growth factors, cytokines and extracellular matrix (ECM) proteins.
We have previously studied the effects of budesonide and formoterol individually however, the effect of their combination on these characteristics of ASM
cells is not known.
Methods ASM cells from asthmatic (n = 6) and nonasthmatic (n = 6) individuals were stimulated with transforming growth factor (TGF) (1 ng/ml) with or
without budesonide (10-8 M) and formoterol (10-10 and 10-8 M) and fibronectin
levels and interleukin-6 (IL-6) release were measured by ELISA. Bronchial
rings from nonasthmatic individuals (n = 2) were incubated with TGF with or
without the drugs and ECM protein expression (fibronectin and collagen I)
measured using immunohistochemistry.
Results In nonasthmatic cells, budesonide alone induced fibronectin deposition whether TGF was present or not. Formoterol decreased fibronectin
induced by TGF and, when combined with budesonide, reversed the increase
in fibronectin. A similar pattern was observed in asthmatic cells, except that
budesonide did not further increase the TGF mediated fibronectin release. As
before [1], IL-6 was induced by formoterol but inhibited by budesonide. TGFinduced IL-6 was inhibited by both drugs and their combination in both cell
types. In bronchial rings the presence of either drug did not affect TGFinduced fibronectin or collagen I.
Conclusion Current asthma therapies are not able to prevent or reverse the
remodelling events regulated by ASM cells.
This work was funded by the NH&MRC Australia.
Reference
Burgess JK et al. 2006. J Allergy Clin Immunol, 118:64957.
Severe Combined Immune Deficiency (scid) spontaneous mutation specifically

impairs differentiation of stem cells into mature lymphocytes. NODCB17prkdscid (known as NOD-scid) lacked NK cells, hence is commonly used
in cell transfer experiments for transferring tissue and haematological
xenografts. The aim of this study was to establish lung inflamamtory model in
NOD-scid strain.
Methods Balb/c and NOD-scid Balb/c mice (n = 8) were exposed to cigarette
smoke for 4 days, 2 and 4 weeks (9 cigarettes/day; 5 days/week). Bronchoalveolar lavage fluid (BALF) and lung tissue were collected for inflammatory
profiling and analysis for cytokines, chemokines and protease expression
and/or activity.
Results NOD-scid have significant accumulation of macrophages in lung
after 4 days, 2 and 4 weeks smoking as compared to no smoke control
(P < 0.001) that was not different to Balb/c (P > 0.05). NOD-scid also have
increased neutrophil number after 2 and 4 weeks smoking (P < 0.001). Even
though myeloid cell differentiation isnt affected by scid phenotype, NOD-scid
have one fold less neutrophil than Balb/c mice (P < 0.001) that is also reflected
in the reduced expression of matrix metalloproteinase-9. Consistent with the
known lymphopenic phenotype, NOD-scid have significant but less lymphocytes recruitment as compared to Balb/c mice after 4 weeks smoking
(P < 0.001) despite the enhanced expression of inteferon inducible protein
10 (lymphocytes specific chemokine) in lung. Both mouse strains showed the
same elevation of net gelatinase and serine protease activity in lung. NODscid mice also demonstrated comparable transcriptional induction of proinflammatory cytokines (TNFa, IL-6), growth factors (GM-CSF, G-CSF) and
chemokines (MCP-1, MIP-2), indicating susceptibility to smoke-induced injury.
Conclusions NOD-scid mice are capable to mount smoke induced inflammatory response. This model may be useful to study localization and role of
immunocytes, including adoptively transfer human cells in the pathogenesis of
COPD.
TP 083
STUDIES OF INTRACELLULAR PAI-2 EXPRESSION

IS COMPLICATED BY ACTIVATION OF THE
INTERFERON RESPONSE
H DAGHER1, J GERN2, R GHILDYAL3, D LI4, P BARDIN1,4
Department of Medicine, Monash University, 2Department of Pediatrics,
University of Wisconsin, Madison, WI 53792, USA, 3Department of
Biochemistry and Molecular Biology, Monash University, Clayton, VIC 3800,
and 4Respiratory and Sleep Medicine, Monash Medical Centre, Clayton, VIC
3168
1
Rhinovirus (RV) is the cause of most common colds and up to 80% of asthma
attacks. In our previous studies, plasminogen activator inhibitor 2 (PAI-2) was
expressed at high levels and was induced in vivo and in vitro by RV infection.
PAI-2 may have antiviral properties suggested by antiviral activity in some
models, high PAI-2 expression levels and further upregulation by RV infection.
Methods To determine whether PAI-2 has antiviral activities following RV
infection, O-Hela, PAI-2 expression-deficient cells were first transfected with
PAI-2 or control genes. This was followed by infection with RV and effects on
viral replication were assessed by RT-qPCR for vRNA and by viral titration for
virus release. IFN expression was assessed by RT-qPCR.
Results IFN-a and -b mRNA expression were induced in response to RV
infection and to PAI-2 expression in cells. PAI-2 expression followed by RV
infection elicited a synergistic response and PAI-2 over-expression reduced
vRNA by >5 fold and viral titre by >3 log (p < 0.05). However, this effect was not
specific to PAI-2, as transfection of cells with control genes/plasmids reduced
viral titre to a comparableextent.
Conclusion Transfection of cells with PAI-2 reduces virus replication by
enhancing IFN activities. However, this effect is not specific as transfection with
control genes induced a similar antiviral effect. Hence, intracellular overexpression of genes as a means of evaluating viral replication may be influenced by switching on innate immune responsesduring the process of
transfection.
Funded by NHMRC (Aust) CJ Martin Fellowship ID 284400.
TP 084
DENDRITIC CELL MIGRATION INDUCED BY KININNS IN

ASTHMATICS AND NON-ASTHMATICS
M FOGEL-PETROVIC, S SHARMA, C BERTRAM, S BALTIC,
P THOMPSON
Lung Institute of Western Australia, Centre for Asthma, Allergy & Respiratory
Research, University of Western Australia, Nedlands, WA 6009
Introduction We reported that dendritic cells (DC) express both kinin B1 and
B2 receptors and that bradykinin (BK), acting through B2R increases concentration of intracellular Ca2+ and mediates immature DC (iDC) migration. Furthermore, we hypothesized that lys-des[Arg9]-BK, which is produced during
inflammation play a role in mature DC (mDC) migration and that both receptors
could be down-regulated and DC-migration decreased with HMG-CoA reductase inhibitors (statins). We also compared the migration index of DC from
healthy and asthmatic subjects.
Methods DC were derived from PBMC isolated from the blood of healthy and
asthmatic subjects. B1R and B2R expression was assessed by confocal microscopy and RT-PCR. Migration of Mo-DC was assessed in transwell chambers.
Chemoattractant used: BK, Lys-des[Arg9]-BK, RANTES and CCL19.
Results Lys-des[Arg9]-BK, which is produced during inflammation, acts
through the B1 receptor (B1R) and play a role in mDC migration, which is
decreased when DC are pre-incubated with kinin. Lovastatin reduced kininsmediated migration by ~50% and chemokine-mediated migration up to ~65%.
Interestingly, DC isolated from asthmatics showed greater motility and
increased sensitivity to inhibition by statin (up to 77%).
Conclusions Kinins may play a crucial role in the regulation of DC migration
in vivo. BK attract migration of iDC, whereas the increased formation of Lysdes[Arg9]-BK in inflammation may decrease the migration of mDC toward
certain chemoattractants and to the draining lymph nodes. Statins, which
down-regulate expression of kinin and chemokine receptors and decrease DC
migration, may potentially be useful as therapeutic agents in inflammatory
diseases.
A46
TP 085
TP 086
THE ROLE OF THE HEDGEHOG PATHWAY IN THE

DEVELOPMENT OF MALIGNANT MESOTHELIOMA (MM)
EFFECT OF STEROIDS ON TGF-1-INDUCED EPITHELIAL TO

MESENCHYMAL CELL TRANSITION (EMT)
L GENOVESI1, C THOMAS2, SE MUTSAERS1,2

Lung Institute of Western Australia, and 2PathWest Laboratory Medicine WA
T HARRIS, M SCHULIGA, A STEWART

Department of Pharmacology, University of Melbourne, Parkville, Vic. 3010
Malignant Mesothelioma (MM) is an aggressive tumour predominantly of the

pleura with a latency period of up to 40 years and is associated with poor
prognosis. Current research is focusing on the molecular mechanisms underlying MM development to identify new targets for therapy, with increasing
evidence pointing to the role of developmental pathways. The Hedgehog (HH)
signaling pathway is significant in a variety of tumours, with a high incidence of
mutations resulting in its constitutive activation, further depicting a clear role for
HH signaling. The role of HH signaling in MM and whether or not an aberration
in this pathway plays a role in the molecular pathogenesis of MM remains to be
determined. We hypothesize that the development of MM is determined, at
least in part, upon HH signaling. To test this hypothesis, the mRNA expression
levels of key HH signaling pathway proteins SHH, IHH, PTCH, SMO, GLI1 by
real time PCR in 2 human primary mesothelial cell cultures and 5 human MM
cell lines. All cells expressed mRNA for PTCH, SMO and GLI1. IHH was not
expressed. SHH was only expressed in one human MM cell line, suggesting
that in MM, HH pathway activation is likely to be predominantly paracrine
driven. In addition, there was significantly higher mRNA levels of most
HH-associated genes in malignant compared with normal mesothelial cells.
Interestingly, the downstream molecule GLI1 was expressed at significantly
high levels in human MM cell lines LO and NO. LO was the only cell line to
express SHH mRNA. NO did not express SHH but produced the highest levels
of transforming growth factor beta (TGF-b), a cytokine considered to be important in MM growth and the only other molecule shown to stimulate GLI expression independent of SMO. The interaction between the HH and TGF-b
signalling pathways are incidently being investigated. In conclusion, the
elevated mRNA levels of the HH signalling pathway components in MM cells is
strong evidence of the potential role of HH signalling in MM.
One of the pathological findings in idiopathic pulmonary fibrosis (IPF) is the

presence on fibroblastic foci comprising cells which exhibit mesenchymal phenotypic features such as myofibroblast-like morphology, increased aSMA
expression and collagen deposition. Currently steroid treatment in IPF has
shown limited efficacy. The cellular origins of these mesenchymal cells remain
unclear, but evidence from other studies suggests that epithelial cells may
undergo a transition to a mesenchymal cell phenotype (EMT). Transforming
growth factor has been implicated in promoting this EMT. In this study we
have induced a morphological change in A549 cells using TGF-1 and
assessed the influence of glucocorticoids, and the changes to the extracellular
environment of the cells, on EMT.
Methods A549 cells were grown on uncoated plastic cultures plates or those
coated with monomeric or fibrillar collagen and treated with 200500 pM TGF1. The influence of the glucocorticoid, dexamethasone (Dex, 11000 nM), or
collagen type, on EMT was assessed by microscopy, RT-PCR and Western
blotting for markers of myofibroblast phenotype.
Results TGF-1 induced an increase in mRNA expression of aSMA (1.5
fold), collagen (7.0 fold) and fibronectin (2.0 fold). Dex (100 nM) partially
inhibited the expression of collagen, but had no effect on aSMA levels.
However, Dex (100 nM) reduced aSMA and CTGF protein levels. Dex
(100 nM) also prevented the TGF-1-induced morphological changes, regardless of ECM matrix.
Conclusion Glucocorticoids appear to control some of the EMT phenotype
changes induced by TGF-1. However, the inability to fully inhibit these
changes may contribute to the resistance of IPF to glucocorticoids. The extracellular environment may also play a role in the development of fibroblastic foci
and their pharmacological responses.
TP 087
THE COLLECTIN PATHWAY AS A POSSIBLE TARGET

FOR THE PRO-PHAGOCYTIC EFFECTS OF AZITHROMYCIN
ON ALVEOLAR MACROPHAGES IN COPD
S HODGE, J AHERN, G MATTHEWS, G HODGE, H JERSMANN,
MD HOLMES, PN REYNOLDS
Research Laboratory, Hanson Institute, Adelaide
Defective alveolar macrophage (AM) phagocytic function in the airway may
perpetuate inflammation via secondary necrosis of uncleared apoptotic cells in
COPD. We have previously reported that low-dose azithromycin improved
macrophage function in vitro, although the mechanisms for this effect were not
identified. We explored the possible role of the collectin pathway in the
azithromycin-mediated improvement in phagocytosis as well as possible
defects in this pathway in COPD subjects.
Methods (1) Mannose binding lectin (MBL), mannose receptor (MR), surfactant protein D (SP-D) were measured in COPD subjects and controls. (2) The
in vitro effects of addition of rhMBL, and blocking MR with a specific antibody,
on AM phagocytic ability were assessed. In vitro effects of azithromycin on AM
expression of MR were also investigated. (3) Azithromycin (250 mg orally 2
weekly/12 weeks) was administered to 11 COPD subjects. Bronchoscopies
were performed prior to and 12 weeks following therapy. Ex vivo assessments
included AM phagocytic ability, levels of MBL, SP-D and MR and apoptosis of
bronchial epithelial cells.
Results AM MR expression and levels of MBL and SP-D were significantly
reduced in COPD subjects vs controls. Azithomycin (500 ng/ml) increased MR
expression by 31% in vitro. rhMBL induced a dose-dependent increase in AM
phagocytic ability (up to 148%). Blocking MR significantly decreased AM
phagocytic ability by 60%. In COPD patients following azithromycin therapy, we
observed improved AM phagcocytic ability, increased levels of MR and reduced
levels of bronchial epithelial cell apoptosis.
Conclusions These findings strongly implicate the MR in both the defective
phagocytic function of AM in COPD and as a target for the azithromycinmediated improvement in phagocytic ability.
TP 088
INCREASED INTRACELLULAR LEUCOCYTE

PRO-INFLAMMATORY CYTOKINE/CHEMOKINE
EXPRESSION IN OBSTRUCTIVE SLEEP APNEA
IS CPAP THERAPY WORKING?
G HODGE1,2, K SRIRAM1, S HODGE1, M HOLMES1, P REYNOLDS1
Department of Thoracic Medicine, Royal Adelaide Hospital, Adelaide,
and 2Haematology Department, Womens and Childrens Hospital,
North Adelaide
Obstructive sleep apnea (OSA) is associated with hypoxia and increased

cardiovascular morbidity. T cells and monocytes play a significant role in
atherogenesis via cytokine production. There have been reports of benefits of
continuous positive airway pressure (CPAP) therapy in OSA. The purpose of
this study was to characterize leucocyte inflammatory cytokine/chemokine
production by T cells and monocytes in a group of OSA patients and to
investigate the therapeutic effects of CPAP therapy.
Methods A comprehensive range of intracellular T-cell and monocyte proand anti-inflammatory cytokines/chemokines was investigated in peripheral
blood from 5 OSA patients and 5 aged-matched control subjects (with no
evidence of sleep problems) using multiparameter flow cytometry. OSA
patients were again studied following 7 days of CPAP therapy.
Results In OSA patients there was an increase in intracellular T-cell IFNg and
TNFa production but no change in IL-2, IL-4 or TGFb compared with control.
There was an increase in intracellular monocyte IL-1a, IL-8, TNFa, MCP-1 and
MCP-3 in OSA patients but no change in IL-10 or IL-12. Following CPAP
therapy, T-cell IFNg and TNFa production returned to normal levels. However,
although intracellular monocyte cytokine/chemokine production was decreased
following CPAP, levels were significantly elevated compared with control.
Conclusions OSA is associated with increased intracellular proinflammatory cytokine/chemokines, many of which are increased in atherosclerotic plaques. Although one week of CPAP therapy resulted in amelioration of
T-cell pro-inflammatory cytokines, longer CPAP use or alternative therapy may
be required to reduce monocyte pro-inflammatory mediators associated with
atherosclerosis in patients with OSA.
TP 089
HUMORAL IMMUNITY TO NONTYPEABLE HAEMOPHILUS

INFLUENZAE
A47
TP 090
THE ROLE OF THE IL-6 FAMILY OF CYTOKINES IN

BLEOMYCIN-INDUCED LUNG FIBROSIS AND
PROLIFERATION
P KING1,2, J NGUI1, M FARMER2, P HOLMES2, S HOLDSWORTH1

Monash University Department of Medicine, and 2Respiratory Medicine,
Monash Medical Centre/Southern Health, Clayton, VIC, 3168
Nontypeable Haemophilus influenzae (NTHi) is a major cause of respiratory

disease, particularly chronic bronchitis. It almost never causes systemic infection and has been shown to be capable of living intracellularly. The investigators have previously described a high prevalence of seroconversion to NTHi in
both controls and bronchiectasis subjects. The aim of this study was to assess
whether humoral immunity to NTHi may have a role in preventing systemic
infection.
Methods Serum was obtained from 10 control subjects and 10 bronchiectasis patients (with chronic NTHi infection). All 20 serum samples had detectable
antibody to NTHi. Ten different isolates of live NTHi were then incubated with
the 10% serum for 2 hours and the effect of killing assessed by the number
of colony forming units (compared with control). The effect of complement
depleted serum and rabbit serum (which lacked antibody to NTHi) was
assessed. The effect of serum killing of NTHi was also assessed by electron
microscopy.
Results The results demonstrated that all 10 strains of NTHi were killed by
the 20 different subject serum specimens; all 200 experiments demonstrated
significant killing of NTHi (with a mean reduction of >99.9% after 2 hours
compared to control). C5, 6 and 8 depleted serum did not kill NTHi but when C5
C6 and C8 was added to the depleted serum >99.9% reduction in NTHi
numbers occurred suggested that killing was mediated through the terminal
attack complex of complement. Electron microscopy demonstrated the presence of pores in bacterial membranes after the addition of serum confirming the
role of the terminal attack complex. Rabbit serum did not have any effect on
NTHi numbers. Blockage of the classical complement pathway by EGTA abolished serum killing.
Conclusion NTHi is very susceptible to extracellular killing by antibody and
the terminal attack complex of complement. The results may explain why NTHi
rarely causes systemic infection and the potential role of intracellular survival.
HL LAU1, R ODONOGHUE1, M ERNST2, D KNIGHT3, S MUTSAERS1,4

Lung Inst. of WA, 2Ludwig Inst. for Cancer Research, Victoria, 3University of
British Columbia, Vancouver, Canada, and 4Pathwest Laboratory Medicine
WA
1
Gp130 has been associated with the progression of fibrosis especially in

patients with idiopathic pulmonary fibrosis (IPF). Gp130 is the common subunit
of the receptor complexes for the IL-6 family of cytokines including IL-11 and
oncostatin M (OSM), where gp130-mediated signalling leads to activation of
the ERK or STAT pathways. We have previously demonstrated exaggerated
gp130-STAT signalling to be fundamental to the development of pulmonary
fibrosis in a murine model of bleomycin-induced lung fibrosis. The aim of this
study was to elucidate the role of the IL-6 cytokine family in the development
of pulmonary fibrosis by identifying which IL-6 family cytokines regulate fibrosis
in bleomycin treated mice, and determine the effects of these cytokines on cell
function. Bleomycin (0.05 U/mouse) or control saline was administered intranasally to wildtype mice (wt), genetically engineered mice containing point
mutations to prevent gp130 ERK signalling (gp130757F) or gp130 STAT signalling (gp130DSTAT), and duel IL-6 and IL-11 a-receptor knockout mice (IL-6-/-;IL11aR-/-). The effect of bleomycin on collagen production was examined in lung
tissue 30 days post treatment by HPLC. There was a significant increase in
collagen levels in bleomycin treated wt lungs which was further increased in
gp130757F lungs. The lungs of gp130DSTAT and IL-6-/-;IL-11aR-/- mice were
protected from fibrosis suggesting that gp130-STAT signalling is important in
inducing lung fibrosis which may be mediated through IL-6 and/or IL-11. Cell
proliferation was examined in lung fibroblasts isolated from wt, gp130DSTAT and
gp130757F mice. IL-6, IL-11 and OSM were significantly mitogenic for gp130DSTAT
cells but not for wt or gp130757F cells, reflecting different responses to the
different signalling pathways. Changes in cytokine profiles are currently being
examined in lung tissue and serum of control and bleomycin treated mice 030
days after treatment. In conclusion, IL-6 and IL-11 are likely to play a role in
bleomycin-induced fibrosis via the gp130-STAT-mediated pathway, however
this may not be due to regulation of proliferation induced by these cytokines.
TP 091
ASYMMETRIC DIMETHYLARGININE AND NITRIC OXIDE

METABOLISM IN A549 AIRWAY EPITHELIAL CELLS
EXPOSED TO CIGARETTE SMOKE AND AN ANTIOXIDANT
J LIU1,2, J WANG3, AS SIM3, DEL WILCKEN3, S CHOW4, DH YATES5,
PS THOMAS1,2,4
1
Prince of Wales Clinical School, Faculty of Medicine, UNSW, 2Department
of Respiratory Medicine, 3Cardiovascular Genetics Laboratory, Prince of
Wales Hospital, 4School of Medical Sciences, Faculty of Medicine, UNSW,
and 5Department of Thoracic Medicine, St Vincents Hospital, Darlinghurst,
NSW 2010
NG, NG-dimethyl-L-arginine (asymmetric dimethylarginine, ADMA) is an endogenous analogue of arginine and competitively inhibitis nitric oxide synthase
(NOS) activity, which is altered by smoking. N-acetylcysteine (NAC), a mucolytic and antioxidant, has been demonstrated to be capable of inhibiting
cigarette smoke solution (CSS) induced NO release from mast cell. It was
hypothesized that CSS would either inhibit NO generation by increasing ADMA
levels or increase NOx levels via an oxidation pathway, which could be inhibited by the antioxidant.
Methods A549 cells were cultured in F-12K Medium (Gibco, USA) with
control medium or 1.0% CSS or 0.8 mM NAC with CSS. Baseline L-arginine,
NOx and ADMA levels were measured in the media. Conditioned media were
then sampled at 1 hr, 6 hrs, 24 hrs, 48 hrs and 72 hrs after incubation. Univariate analysis was used to calculate p values.
Results CSS induced significantly higher NOx levels when compared with
control media (p < 0.0005). NAC pre-treatment partially reversed this effect
(p = 0.009). Percentage increase in ADMA levels per million cells was significantly higher in the CSS conditioned media compared with control media
(p = 0.02) while NAC pre-treatment did not affect ADMA media levels
(p = 0.07).
Conclusion CSS exposure generated significantly higher levels of NOx
when compared with control media which could be partially reversed by NAC
pre-treatment. ADMA levels were increased after CSS exposure. This suggests
that both NOx and ADMA were generated in response to CSS reflecting an
overall activation of the NO pathway and its regulatory mechanisms. This
activation is via an oxidative stress pathway and not via the NOS pathway.
TP 092
INDUCTION OF AN OSTEOBLAST PHENOTYPE IN MOUSE

MALIGNANT MESOTHELIOMA CELLS IN VITRO
B PEDERSEN1, SM LANSLEY1, RG SEARLES1, C THOMAS1,
S WILKOSZ1, PJ THOMPSON1, G STERRETT2, CM PRLE4,
SE MUTSAERS1,2
1
Lung Institute of Western Australia, 2PathWest Laboratory Medicine WA,
4
Telethon Institute for Child Health Research and Centre for Child Health
Research, UWA, Perth, Western Australia
Malignant mesothelioma (MM) is largely an untreatable tumour predominantly
of the pleura with a latency period of up to 40 years. A better understanding of
the biology of this disease is needed for the development of novel treatments
to improve survival. Features of osteogenic tissue have been observed in
tumours within a subset of patients with MM and also in experimental mouse
models, however its origin is unknown. We hypothesize that bone formed
within the tumour is derived from MM cells with the capacity to differentiate into
cells of an osteoblastic lineage. To test this hypothesis, murine MM cells were
applied to a functional in vitro assay of bone formation. Four MM cell lines were
grown in osteogenic medium (OM) to induce bone formation as well as standard culture medium. The expression of alkaline phosphatase, an early osteoblast phenotype marker, was present by 15 days in all cell lines examined with
variability in staining patterns observed between cell lines. Von Kossa staining
of cells demonstrated mineralized bone nodules in a single cell line, providing
conclusive functional evidence of bone formation in these cells. All murine MM
cells differentially expressed mRNA and protein of several key osteoblast
markers, including core binding factor alpha-1, osteonectin, osteopontin and
bone sialoprotein, as assessed by RT PCR and Western blotting respectively.
In conclusion, there is strong evidence to suggest that mouse MM cells have
the capacity to differentiate into cells of an osteoblast lineage and that this is
the likely source of bone found in MM tumours.
A48
TP 093
TP 094
HUMAN AND MOUSE TUMOURS EXPRESS TLR7: EVIDENCE

FOR IN-VIVO SENSITIVITY TO LOCAL TREATMENT WITH
IMIQUIMOD
GLUTATHIONE PEROXIDASE-1 PROTECTS AGAINST

CIGARETTE SMOKE-INDUCED LUNG INFLAMMATION
S BROOMFIELD, R VAN DER MOST, A PROSSER, S MAHENDRAN,

B ROBINSON, A CURRIE
Tumour Immunology Group, School of Medicine and Pharmacology,
University of Western Australia
Mimicking viral infection by application of various toll-like receptor ligands has

shown clinical promise in the treatment of persistent viral infections and more
recently with malignant tumours. Commercially available toll-like receptor 7
ligands (TLR7L), such as those of the Imidazoquinoline family have been
applied clinically for the treatment of a number of conditions including basal cell
carcinoma and HPV-induced genital warts. These compounds are known to
retard tumour growth indirectly by promoting activation and migration of DCs,
leading to a strong Th1 cellular response, and directly via release of proinflammatory cytokines and promotion of tumour cell apoptosis. Malignant
mesothelioma (MM), an aggressive tumour with a mean survival of 9 months,
is highly resistant to chemotherapy, radiotherapy and surgery and is therefore
an interesting candidate for immunotherapy in the form of TLR7 ligand treatment. Whilst TLR7 is known to be selectively expressed in immune cells and its
relative expression low amongst other cell and tissue types in mammals, its
expression on tumour cells and the consequences of such expression on
tumour growth are unknown. Here we describe the presence of TLR7 (mRNA
and protein) directly in a range of different tumours, including several murine
and human MM cell lines. In-vivo Imiquimod treatment of the murine MM
tumour AB1-HA significantly retarded tumour growth in all treated mice. Nude
mouse and depletion studies revealed a significant role for CD8 T cells and NK
cells in the local anti-tumour immune response. A systemic response was
observed from in-vivo CTL analysis and Ly6A/E staining of lymphocytes, suggesting the potential of a systemic vaccine response. Interestingly, TLR7L
synergized with systemic anti-CD40 treatment to significantly enhance local
tumour resolution. Inducing inflammation and apoptosis by targetting TLR7 in
the tumour environment may be a useful therapeutic approach for the treatment of malignant mesothelioma.
R VLAHOS1, S BOZINOVSKI1, P HERTZOG2, P CRACK1, G ANDERSON1

The University of Melbourne, Parkville, VIC 3010 and 2Monash Institute of
Medical Research, Clayton, VIC 3168
Reactive oxygen species (ROS) produced during the innate immune response
are important agents of anti-pathogen defense but may also cause oxidative
lung damage. Glutathione peroxidase-1 (gpx-1) is a detoxifying enzyme that
may protect lungs from such damage.
Methods Wild-type (WT) or mice deficient in glutathione peroxidase-1 (gpx1-/-) were placed in a perspex chamber and exposed to cigarette (cig) smoke
generated from 9 cigs per day for 4 days. On the fifth day, mice were killed,
the lungs lavaged with PBS and then harvested for proteomic and genomic
analysis.
Results WT mice exposed to cig smoke for 4 days had significantly more
macrophages (3.1 0.1(SEM) 105) and neutrophils (4.9 0.4 105) than
sham-exposed mice (2.2 0.2 105 and 0, respectively) (n = 6, p < 0.05).
However, gpx-1- mice exposed to cig smoke had significantly greater
macrophages (5.4 0.3 105) and neutrophils (1.2 0.1 106) than smokeexposed WT mice (n = 6, p < 0.001). Macrophage and neutrophil numbers in
sham-exposed gpx-1-/- mice (1.7 0.3 105 and 0.5 0.4 103) were similar
to those of sham-exposed WT mice (2.2 0.2 105 and 0). In addition, we
found that BALF of gpx1-/- mice exposed to cig smoke had an increased
proteolytic burden compared with smoke-exposed WT mice as assessed by
zymography and net gelatinase activity assay.
Conclusions These data suggest that gpx-1 protects the lung from cigarette
smoke-induced inflammation and that targeting gpx-1 may have therapeutic
utility in inflammatory lung diseases where cigarette smoke plays a role.
Funded by NHMRC.
COPD SIG: Poster Session 1

TP 096
TP 095
IMPAIRED RELEASE OF INFLAMMATORY MEDIATORS FROM

COPD BRONCHIAL EPITHELIAL CELLS (BECs) IN RESPONSE
TO RHINOVIRUS(RV) AND LIPOPOLYSACCHARIDE (LPS)
OPINIONS AND ATTITUDES TO A 12-MONTH EXERCISE

PROGRAM IN COPD PATIENTS
LISSA SPENCER1,2,3, JENNIFER ALISON1,2, ZOE MCKEOUGH1
The University of Sydney, Sydney, Australia 2006, 2Royal Prince Alfred
Hospital, Sydney, Australia 2050 and 3NSW Dept of Health, North Sydney,
Australia 2059
1
1
PAB WARK , B DAVIES , K PARSONS , T GRISSELL

Department of Respiratory and Sleep Medicine John Hunter Hospital, and
2
School of Medicine and Public Health, University of Newcastle, Newcastle,
Australia
The BECs from subjects with chronic obstructive pulmonary disease (COPD)
are exposed to frequent infectious and inflammatory stimuli. Infection with RV
is known to trigger acute exacerbations and subjects with COPD are particularly susceptible. We hypothesized that exposure of COPD BECs to these
stimuli would alter their response to RV infection.
Methods BEC were obtained by endobronchial brushing from subjects with
GOLD stage 3 COPD (n = 4, all ex-smokers), subjects with mild persistent
asthma (n = 4) and healthy controls (HC, n = 4). BECs were cultured and then
treated with Tumour Necrosis Factor (TNF)a 10 ng/ml or LPS 100 mg/ml for
24 hrs and then infected with RV-43, RV-1B. Response was measured by
release of IL-8, IL-6 and IP-10 mRNA and by ELISA. Virus replication measured by cell titration assay.
Results Infection with both RV strains led to increased release of IL-8 and
IP-10 in all groups. Exposure of HC and asthma BECs to both LPS and TNF led
to increased release of IL-8. In these BECs there was no increase in release of
IL-8 exposed to LPS and TNF and then infected with either RV. BECs from
subjects with COPD released significantly less IL-8 in response to all conditions
and RV infection compared to HCs and asthma. No differences were seen in
RV replication.
Conclusions BECs from subjects with moderate to severe COPD release
less IL-8 in response to RV infection and exposure to LPS and TNFa. Preexposure of BECs for 24 hr to TNF and LPS does not alter this response.
The aim of this study was to determine opinions and attitudes to exercise from
chronic obstructive pulmonary disease (COPD) subjects after completion of a
12-month maintenance exercise program.
Methods Following completion of a 12-month exercise study, which included
a supervised program (Intervention, n = 18) and control group (Control, n = 17),
COPD subjects [mean age (SD): 66 (8); mean FEV1 (% predicted) = 56% (19)]
were asked to complete a questionnaire. The questionnaire included closedended questions using visual analogue scales (100 mm).
Results Data from the questionnaire showed that the intervention group
exercised more regularly during the 12 months [Intervention: 64 mm (23);
Control: 42 mm (24), 95%CI: -35 to -3] and reported more enjoyment at being
involved in the study [Intervention: 96 mm (9); Control: 86 mm (16), 95%CI:
-17 to -2]. Both groups reported that exercise was important [Intervention:
97 mm (5); Control: 92 mm (14), 95%CI: -10 to 3] and their 12-month program
was beneficial in improving some aspect of their life [Intervention: 89 mm (11);
Control: 80 mm (25), 95%CI: -20 to 2]. Both reported that exacerbations were
a barrier to exercise [Intervention: 47 (27); Control: 35 (29); 95%CI: -30 to 8]
and that support was important for adherence to exercise [Intervention: 97 (6);
Control: 92 (14); 95%CI: -12 to 2].
Conclusion COPD subjects have positive attitudes towards both supervised
and unsupervised maintenance exercise programs and that asking patient
opinion may help health professionals design more acceptable long-term exercise programs.
TP 097
A49
TP 098
END OF LIFE ISSUES AND CHRONIC OBSTRUCTIVE

PULMONARY DISEASE (COPD)PATIENT AND CARER
OPINIONS
END OF LIFE ISSUES AND CHRONIC OBSTRUCTIVE

PULMONARY DISEASE (COPD) MEDICAL AND NURSING
OPINIONS
AL KEAN1, MM ROBERTS1, P DAVIDSON2, SR LEEDER1, TD ROBINSON1

Respiratory Medicine, 2Nursing Research Unit, Westmead Hospital, NSW
2145
MARY ROBERTS, ANNETTE KEAN, TRACEY ROBINSON

Department of Respiratory Medicine, Westmead Hospital, NSW 2145
Recent position papers have highlighted the importance of end-of-life discussions with patients with chronic disease. Disease trajectory in COPD is
unpredictable and so many patients miss the opportunity to participate in
end-of-life decision-making
Aims To assess the knowledge of patients with COPD and their carers about
end-of-life decision-making and to determine their desired level of participation.
Methods 4 focus groups of 5 to 9 participants including patients with severe
COPD (FEV1 37 11 %predicted [mean SD]) and carers plus 1 focus group
with carers of recently deceased patients were held over a 12 month period,
using a facilitated interview technique. Transcripts from focus group interviews
were interpreted using thematic content analysis.
Results Patients and carers had surprisingly poor insight into the progressive
nature of COPD and did not consider their disease to be immediately life
threatening. Their experiences in hospital were characterized by feelings of
powerlessness, based on uncertainty about prognosis, poor understanding of
hospital staff hierarchy (different messages from multiple, often unidentified
staff members) and the use of jargon particularly Not For Resuscitation (NFR).
Disturbingly, patients previously involved in resuscitation discussions in hospital felt bullied into accepting NFR. Some patients associated making end-of-life
plans with withdrawal of treatment and thought that NFR was an irreversible
decision. Patients wished to remain hopeful during a disease exacerbation
rather than discuss matters relating to the end-of-life. Interestingly, the carers
of recently deceased patients described a process whereby the patients initiated treatment withdrawal within 48 hours of death.
Conclusions Patients are unaware of or reluctant to consider the progressive nature of their disease and prefer to remain hopeful about current and
future life-saving therapies. Patients and carers need more education about
COPD prognosis before they will be able to acknowledge the importance of
participating in end-of-life planning.
NSW Health guidelines and position papers emphasize the importance of

end-of-life discussions with patients with chronic disease within the last 612
months of life. Patients with chronic obstructive pulmonary disease (COPD) are
frequently hospitalized with acute exacerbations but disease trajectory is
unpredictable and so determining the timing and venue of end-of-life discussions may be difficult.
Aims To examine the opinions and practice of hospital-based medical and
nursing staff regarding end-of-life discussions with patients with COPD.
Methods 4 focus groups of 4 to 7 participants were held using a facilitated
interview technique. Each group consisted of 1 professional group ie Respiratory physicians (RP), advanced trainees (AT), junior resident medical officers
(JRMO) and senior respiratory nurses (SRN). Transcripts from focus group
interviews were interpreted using thematic content analysis.
Results Common themes identified by all groups included patients poor
understanding of COPD, especially prognosis and treatment options, a reluctance to document and communicate end-of-life discussions and dealing with
cultural diversity. Both RP and AT identified the unpredictable course of COPD
and that Not For Resuscitation (NFR) was often interpreted as Not for Treatment. Only RP identified that most patients did not want to discuss end-of-life
issues and preferred to remain positive. RP saw themselves as advocates for
life-saving therapy for patients. SRN felt disempowered from the decision
process and felt they were inflicting distressing treatments on patients. RP
questioned the need to initiate end-of-life discussions early, in conflict with
other groups, who thought it was important for RP to discuss these issues
sooner rather than later with patients, and that the hospital setting was not an
appropriate venue.
TP 099
BILEVEL VENTILATION DURING EXERCISE IMPROVES

EXERCISE CAPACITY IN ACUTELY UNWELL PATIENTS WITH
HYPERCAPNIA
TP 100
PUBLISHED REFERENCE EQUATIONS UNDERESTIMATE 6

MINUTE WALK DISTANCE FOR MIDDLE AGED AND ELDERLY
AUSTRALIANS
C MENADUE1,2, ER ELLIS2, AJ PIPER1,2, D FLUNT1, JA ALISON1,2

Department of Respiratory and Sleep Medicine, Royal Prince Alfred
Hospital, NSW 2050, and 2Discipline of Physiotherapy, The University of
Sydney, NSW 2141
Exercise rehabilitation may be delayed in acutely unwell patients with hypercapnic respiratory failure and severe dyspnoea. Non-invasive ventilation (NIV)
during exercise can reduce dyspnoea and improve endurance time in patients
with stable severe chronic obstructive pulmonary disease. However the effect
of NIV during exercise in the acute care setting is unknown. Therefore, a
randomized crossover study with repeated measures was conducted to determine the effect of NIV on exercise capacity and dyspnoea in acutely unwell
patients with hypercapnic respiratory failure during walking and unsupported
arm exercise (UAE).
Method Twenty participants (mean SD PaCO2 60 10 mmHg) performed
six minute walk tests (6MWT) and seventeen participants (PaCO2
59 10 mmHg) performed UAE tests in random order with NIV and oxygen
(O2), and O2 alone.
Results
6MWT Total
distance (m)
Time to 1st rest (s)
Distance to 1st
rest (m)
Isotime dyspnoea
(Borg)
UAE Endurance
time (s)
Isotime dyspnoea
(Borg)
O2
NIV and O2
Mean (95% CI)
180 89
222 114
42 (1578)
0.004
197 (144360)
154 95
360 (249360)
212 120
58 (2096)
0.011
0.005
4.0 (3.05.0)
2.0 (1.04.0)
0.019
156 (87325)
189 (92394)
0.024
3.0 (2.57.5)
3.0 (2.05.0)
0.066
Conclusions NIV and O2 during exercise can allow acutely unwell patients
with respiratory failure to exercise for longer and may reduce dyspnoea compared to exercise with O2. Further research is needed to determine if NIV
during exercise rehabilitation can prevent deconditioning or cause a sustained
improvement in exercise capacity in acutely unwell patients with respiratory
failure.
Support Physiotherapy Research Foundation, APA.
S JENKINS1,2,3, N CECINS1,2,3, B CAMARRI1,2, C WILLIAMS1,2,

P THOMPSON1,2, P EASTWOOD1,4,5
1
Institute of WA, Nedlands, WA 6009, Departments of 3Physiotherapy and
4
Pulmonary Physiology, Sir Charles Gairdner Hospital (SCGH), Nedlands,
WA 6009, and 5School of Anatomy and Human Biology University of WA,
Crawley, WA 6009
Ethnic and regional differences in 6 minute walk distance (6MWD) exist. This
study developed regression equations to predict 6MWD in healthy Caucasian
Australians aged 4585 yrs and to compare measured 6MWDs with predicted
6MWDs obtained from studies in North America1,2,3 and Europe4.
Methods 109 adults (48 males) aged 62.1 8.3 yrs completed two 6MWTs.
Measurements of height, weight and habitual physical activity were obtained.
Results 6MWD (maximum of 2 tests) was 682 73 and 643 70 m in
males and females respectively (p < 0.01). Age, height and %predicted
maximum heart rate (%predHRmax) achieved during the 6MWT were independent contributors (p < 0.05) to 6MWD. In females, there was an inverse
relationship between BMI and 6MWD (r = -0.39, p < 0.01). Regression equations were developed using all significant variables and only age and anthropometric data (Table).
Predicted 6MWD (m) height (ht) in cm; M: males; F: females
M
M
F
F
748 (6.32 age) + (0.64 ht) + (2.69%predHRmax)

867 (5.71 age) + (1.03 ht)
541 (3.81 age) + (1.80 ht) (6.92BMI) + (2.41%predHRmax)
525 (2.86 age) + (2.71 ht) (6.22BMI)
R2
0.61
0.40
0.58
0.43
Predicted 6MWDs obtained from other studies underestimated 6MWD by

mean (95% CIs, m) 114 (98,130)1, 31 (14,47)2, 186 (156,216)3 and 27 (12,43)4
(females) and 107 (86,128)1 and 182 (153,211)3 (males) (p < 0.001).
Conclusions These findings highlight the importance of population specific
equations to predict 6MWD.
Supported by NHMRC and SCGH.
References
1. Am J Crit Care Med 1998;158;13847.
2. J Cardiopul Rehabil 2001:21:8793.
3. Chest 2003; 123:38798.
4. ERJ 1999;14:2704.
A50
TP 101
TP 102
INFLUENCE OF DISABILITY ON CHANGES IN 6 MINUTE WALK

DISTANCE FOLLOWING A FAMILIARIZATION TEST AND
TRAINING
ENDOTOXIN LEVELS ARE HIGHER IN THE

BRONCHOALVEOLAR LAVAGE OF EX-SMOKERS COMPARED
TO NEVER SMOKERS
S JENKINS1,2,3, P EASTWOOD1,4,5, K HILL1,2, N CECINS1,2,3

School of Physiotherapy, Curtin University of Technology, WA 6845, 2Lung
Institute of Western Australia, Departments of 3Physiotherapy and
4
Pulmonary Physiology, Sir Charles Gairdner Hospital, and 5School of
Anatomy and Human Biology, University of Western Australia, WA 6009
PETER AB WARK1, REBECCA OLDHAM1, JODIE L SIMPSON3,

JEROEN DOUWES2
1
Department of Respiratory and Sleep Medicine John Hunter Hospital and
School of Medicine and Public Health, University of Newcastle, Newcastle,
Australia, 2Centre for Public health Research, Massey University, Auckland,
New Zealand, and 3NHMRC Centre for Respiratory and Sleep Medicine, The
University of Newcastle
In COPD the 6 minute walk distance (6MWD) is known to increase with test
repetition (familiarization) and in response to exercise training. It is unknown
whether the magnitudes of these increases are related to the degree of disability of the individual patient.
Methods 6MWD was measured twice before and once after an 8 week
out-patient exercise program in 121 patients (82 males) aged 678.6 yrs,
FEV1 3715% predicted (meanSD) with stable COPD. The changes in
6MWD following a familiarization test and following training were compared
between patients grouped according to their degree of disability (defined as the
pre-training 6MWD [best of 2 tests] expressed as %predicted 6MWD).
*p < 0.05 Gp 3 vs Gp 1.
Results
Familiarization
Test 1 (m)
Test 2 (m)
Change m (%)
Post-training
6MWD (m)
Change m (%)
Gp 1 <60%
n = 32
Gp 2 6080%
n = 55
Gp 3 >80%
n = 34
275 67
418 49
514 56
301 50
28 31 (12 15)
351 66
455 51
37 31 (9 8)
495 57
559 57
44 29* (9 6)
582 64
48 51 (18 20)
39 42 (8 9)
23 22* (4 5*)
Endotoxin is a stimulant of the innate immune system and is a major component of cigarette smoke. Smokers have evidence of increased airway neutrophils and inflammation. We hypothesized that endotoxin levels would be higher
in the bronchial lavage (BL) of subjects who were former smokers and subjects
with chronic obstructive pulmonary disease (COPD).
Methods Subjects were all ex-smokers for at least 5 years (n = 10, 5 COPD,
5 healthy controls) or never smokers (n = 12, 6 asthma, 6 healthy controls). BL
was collected and analysed for cell count and differential, culture for microbiology. The supernatant was analysed for IL-8 by ELISA and endotoxin by
quantitative kinetic LAL assay.
Results Median endotoxin levels were significantly higher in ex-smokers 101
compared to never smokers 6.3 U/ml (p < 0.001). There were no differences
between subjects with COPD and HS. Subjects with COPD had higher median
endotoxin levels (80 U/ml), compared to asthma (5.2 U/ml) and HC (6.3 U/ml,
p = 0.03). There was no correlation between endotoxin levels and BL total cell
count, neutrophils (%) or FEV1 % predicted. There was a strong correlation
with previous packet years smoked and endotoxin levels (r = 0.72, p < 0.01).
Conclusions BL endotoxin levels are higher in ex-smokers, including subjects with COPD. Despite this there is no relationship to increased neutrophilic
inflammation.
Conclusions Before training, 6MWD increases following a familiarization

test irrespective of the level of disability. The magnitude of this increase is
similar in all groups when normalized for their pre-training 6MWD. Following
training, the increase in 6MWD is greatest in patients with the greatest disability
(lowest pre-training 6MWD). In less disabled patients, the relatively smaller
increase in 6MWD following training may reflect an inability to further increase
stride length, thereby reducing the responsiveness of the 6MWT in this group.
Supported by NHMRC.
TP 104
TP 103
LOW DOSE AZITHROMYCIN IMPROVES EFFEROCYTOSIS

(APOPTOSIS AND MACROPHAGE FUNCTION) AND REDUCES
INFLAMMATION IN COPD PATIENTS
SANDRA HODGE, JESSICA AHERN, GEOFF MATTHEWS, GREG HODGE,
HUBERTUS JERSMANN, MARK D HOLMES, PAUL N REYNOLDS
COPD is associated with inflammation associated with ineffective repair of the
injured epithelium and loss of structural integrity. We have shown that these
changes may result from dysregulated efferocytosis (increased apoptosis of
bronchial epithelial cells and defective clearance of these cells by alveolar
macrophages (AM)). We have also reported that azithromycin, at subbactericidal dose, improved AM phagocytic function ex vivo.
Methods We administered azithromycin at low dose (250 mg/ twice weekly
for 12 weeks) to 10 COPD subjects (7 male, age: 62 8 yr, 5 current/ 5
ex-smokers, FEV1: 63 9% pred, FEV1/FVC: 48 9%). The study was openlabel, uncontrolled and primarily focused on objective biological responses
obtained from the bronchoscopy samples taken. Phagocytic ability of AM (from
BAL), apoptosis of bronchial epithelial cells (from bronchial brushing), markers
of inflammation in blood, BAL and breath condensate (CRP, WCC and inflammatory cytokines), health status (St. Georges respiratory questionnaire), ECG
and lung function were assessed pre and post- administration of azithromycin.
Results Azithromycin significantly improved phagocytic ability of AM
(by 37%) and reduced bronchial epithelial cell apoptosis (by 34%). Antiinflammatory effects of azithromycin included significantly reduced blood WCC
and CRP. There were non-significant reductions in levels of pro-inflammatory
cytokines IL-8, IL-6 and TNF-a in blood, BAL and breath condensate, and a
trend for improved health status.
Conclusions Our findings indicate a novel approach to supplement existing
therapies in COPD that may improve clearance of accumulated apoptotic
material and reduce the risk of secondary necrosis and release of toxic cell
contents that perpetuate inflammation.
THE PREVALENCE OF GASTRO-OESOPHAGEAL REFLUX

DISEASE (GORD) IN COPD AND BRONCHIECTASIS
A LEE1, B BUTTON1,2, L DENEHY1, S ROBERTS3,6, S ELLIS4,
R STIRLING5,6, J WILSON5,6
1
School of Physiotherapy, The University of Melbourne, Vic 3010,
Departments of 2Physiotherapy, 3Gastroenterology, 4Radiology and
5
AIRMED, The Alfred, Vic 3004, and 6Department of Medicine, Monash
University, Vic 3800
Background The prevalence of gastro-oesophageal reflux disease (GORD)
across the disease spectrum in COPD and bronchiectasis is not well described.
The aim of this study was to determine the prevalence of symptomatic and
silent GORD in COPD and bronchiectasis and its effect on lung function and
quality of life (QOL).
Methods Patients and controls completed dual-probe 24 hr oesophageal pH
monitoring, measuring number of reflux episodes (NRE) and % reflux time (RI)
in the proximal and distal oesophagus. Disease severity (lung function and
HRCT scoring) and QOL (Short Form-36 [SF-36], St Georges Respiratory
Questionnaire [SGRQ] and Quality of Life in Reflux and Dyspepsia (QOLRAD)]
were measured.
Results Thirty patients with bronchiectasis (mean [SD] FEV1 74% predicted
[23.4]), 27 with COPD (mean [SD] FEV1 47% predicted [17.4]) and 18 healthy
controls were recruited. The prevalence of GORD in bronchiectasis was 33%;
37% in COPD; 17% in controls. In COPD and bronchiectasis, total NRE and RI
were increased in those with distal and proximal GORD compared to those
without GORD (all p < 0.05). There was no difference in extent or severity of
bronchiectasis in patients with or without GORD (all p > 0.05). In COPD, the
relationship between proximal GORD and FEV1 was small to moderate
(r = 0.383). SGRQ symptom scores were higher in patients with bronchiectasis
with increased RI (p = 0.02). Increased proximal NRE was associated with
reduced physical (p = 0.03) and mental health (p = 0.02) in the SF-36 in COPD.
Conclusions GORD is a co-morbidity in patients with COPD and
bronchiectasis. The impact of GORD on disease severity requires further
evaluation.
Funding source NHMRC, The University of Melbourne, Monash University,
Physiotherapy Research Foundation.
A51
TP 105
TP 106
AGE AND DISEASE RELATED CHANGES IN AIRWAY

INFLAMMATION IN OLDER PEOPLE WITH COPD
MICRORNA EXPRESSION PROFILING OF COPD SEVERITY

USING MICROARRAYS
JODIE L SIMPSON1,2, PHILIP M HANSBRO2, VANESSA M MCDONALD1,2,

PETER G GIBSON1,2
1
NHMRC Centre for Respiratory and Sleep Medicine, University of
Newcastle, NSW, Australia, and 2Centre for Asthma and Respiratory
Disease, Hunter Medical Research Institute, NSW, Australia
SM SAVARIMUTHU FRANCIS1,2, JE LARSEN1,2, NK HAYWARD3, RV

BOWMAN1,2, PV ZIMMERMAN1,2, KM FONG1, IA YANG1,2
1
The Prince Charles Hospital, 2University of Queensland, and 3Queensland
Institute of Medical Research, Brisbane, Australia
Chronic obstructive pulmonary disease (COPD) is prevalent among older

people, however little is known about the influence of ageing on airway inflammation. The aim of this study was to compare airway inflammation in older
people with obstructive airway disease to groups of older and younger healthy
controls.
Methods Participants (>55 years of age) with stable airway disease and
incomplete reversibility (FEV1% predicted <80% and FEV1/FVC < 70%; COPD
n = 71) and healthy controls (n = 45, 35 older >55 years and 10 younger <55
years) were recruited from the Respiratory Ambulatory Care Clinic or by advertisement. Participants underwent a clinical assessment, skin allergy test,
hypertonic saline challenge, sputum induction and gas diffusion studies.
Results Participants with COPD had moderate airflow obstruction (mean
(SD) FEV1% predicted 56 (19)) and 45 (63%) were current or ex-smokers with
a median (IQR) pack year history of 36 (2054) pack years. Ageing was
associated with an increase in airway neutrophils (p = 0.0001). Compared to
older controls, participants with COPD had increased airway eosinophils and
lymphopenia (p = 0.004, p = 0.003 respectively), but no difference in airway
neutrophils.
Conclusion Airway neutrophilia is a feature of ageing and is not further
increased in the presence of COPD. COPD is associated increased numbers
of airway eosinophils with reduced lymphocytes which may impact on the
ability of the immune system to combat infection.
Supported by NHMRC, The University of Newcastle.
Chronic Obstructive Pulmonary disease (COPD) is third leading cause of death

and fourth leading cause of disease burden in Australia. Mechanisms involved
in emphysema severity have not been fully understood. MicroRNAs are noncoding RNAs that regulate gene expression. We hypothesize that microRNA
expression differs between emphysema severity in COPD patients.
Methods miRNA profiling was performed using 15K Agilent Human Oligo
miRNA microarrays on total RNA extracted from non-tumour lung tissue from
30 COPD patients undergoing resection for lung cancer. The miRNAs were
quantile normalized and ANOVA was used to find differentially expressed
genes.
Results Demographic characteristics of the COPD patients (mean (SD))
were age 69 (6) years, FEV1 72 (17) % predicted and FEV1/FVC ratio (<70%).
ANOVA identified 31 miRNAs that were differentially expressed when stratified
into two classes according to KCO % predicted > or <75% (t-test, P < 0.05).
Discussion This miRNA analysis has identified miRNAs that may be important in emphysema severity in COPD patients. Further validation will be performed using qRT-PCR and miRNA assays on the training set and an
independent set, and target prediction and validation.
Supported by NHMRC Biomedical Scholarship and CDA, The Prince Charles
Hospital Foundation, Queensland Clinical Research Fellowship.
COPD SIG: Poster Session 2

TP 108
FORCED EXPIRATORY TIME VARIABILITY AND PREDICTION OF

AIRWAY OBSTRUCTION?
M SWANNEY1, L BECKERT1, J STANTON1, P KELLY1, C FRAMPTON2
Respiratory Physiology Laboratory, Christchurch Hospital, Christchurch,
New Zealand,8140, and 2 University of Otago, Christchurch, New Zealand
8140
TP 107
T HELPER-17 CELL RESPONSES IN CHRONIC OBSTRUCTIVE

PULMONARY DISEASE: A PILOT STUDY
Airway obstruction is defined as a FEV1/FVC ratio below the lower limit of

normal. Airway obstruction may prolong the forced expiratory time (FET).
Method Spirometry results from 467 patients were categorized as obstructive, restrictive or normal. The mean, range and coefficient of variation were
determined for FET in each diagnostic group. Receiver operator characteristic
(ROC) curves were used to determine if FET could predict a low FEV1/FVC.
The number of patients with airway obstruction in five FET groups: <9; 9;
1012; 1314; and >14 seconds were determined.
Results The coefficient of variation was high for all groups. Pair-wise comparisons showed a difference in mean FET between patients with normal lung
function versus those with airway obstruction (p < 0.001). The best cut-point in
the ROC analysis of 9.895 seconds had a sensitivity of 0.66, specificity 0.77
and area under the curve of 0.743 for predicting obstruction.
JENNIFER PERRET1, 2, NICOLE BROOKS1, 2, VASSO APOSTOLOPOULOS2,

CHRISTINE MCDONALD1, DODIE POUNIOTIS2*
1
Institute for Breathing and Sleep (IBAS) and 2Burnet Institute, Austin
Campus, Heidelberg Victoria 3084
FET
mean
(95% CI)
FEV1/FVC
CV%
FET CV%
Normal
Normal
Variant
(FEV1>LLN)
Mild
Moderate
Severe
8.9
(8.59.3)
8.5
(7.99.0)
11.5
(10.312.7)
11.9
(11.112.7)
12.4
(11.613.3)
11.3
(10.512.2)
2.53
2.73
12.3
3.42
11.6
12.4
3.72
13.3
4.29
11.2
7.01
12.4
LLN = Lower limit of normal.

Percentage of patients with Airway Obstruction
T-helper type 1 (TH1) and type 2 (TH2) lymphocyte responses have been well
recognized as being important pathways in inflammation. Recently another
form of inflammatory lymphocyte response has been described, the TH17
pathway. TH17 cells produce cytokines such as IL-17A to clear extra-cellular
bacteria and fungi and have been implicated in autoimmune and chronic
inflammatory diseases. The TH17 response in COPD is unknown.
Methods Subjects were patients with COPD (ex-smokers, FEV1 < 70% predicted who had not had an exacerbation for at least 1 month) and control
subjects (ex-smokers and normal spirometry). Serum samples were obtained
for measurement of C reactive protein (CRP) and IL-17A, the latter measured
using enzyme-linked immunosorbent assay (ELISA). Production of IL-17A by
T-cell subsets was also identified by intra-cellular cytokine staining and measured by flow cytometry.
Results The mean FEV1 of COPD subjects was 42 % predicted (6.1 SEM,
n = 6) and mean FEV1 of controls was 112 % predicted (3.0 SEM, n = 4). The
COPD group had a higher mean level of CRP 9.5 mg/l (3.9 SEM) compared to
the control group mean level of 4.6 mg/l (0.6 SEM). The mean level of the IL-17
in the COPD group as measured by ELISA was 22.3 pg/ml (16.9 SEM, range
087) whilst no IL-17 was measured in any of the control subjects.
Conclusions The findings of this pilot study suggest that IL-17 may be
elevated in association with CRP in stable COPD.
Airway
Obstruction
Restrictive
pattern
100
80
60
40
20
n=204
n=55
n=128
n=36
<9
10 - 12
13 - 14
n=44
>14
Forced Expiratory time (seconds)
Conclusion Although the coefficient

of variation for FET is high we could
distinguish those with airway obstruction based on FET. Further research is
required to investigate if FET has a role
in a screening strategy for predicting
airway obstruction.
A52
TP 109
HOMEBASED SPIROMETRIC AND INSPIRATORY CAPACITY

MEASUREMENTS IN SEVERE COPD ARE REPRODUCIBLE
NICK ANTONIADES1, PETER ROCHFORD1, JEFFREY PRETTO1,
ROB PIERCE1, JANETTE GOGLER2, JULIE STEINKRUG2,
CHRISTINE MCDONALD1
1
Institute for Breathing and Sleep, Department of Respiratory & Sleep
Medicine, and 2Department of Nursing Informatics, Austin Health, Victoria
3084
Remote in-home measurement of a variety of physiological variables (telemonitoring) provides an alternative management approach for patients with
chronic disease. There are few data demonstrating the reproducibility of such
measurements in COPD.
Aim To determine intra-patient reproducibility of unsupervised spirometry,
including measurement of inspiratory capacity (IC) in the home in severe
COPD.
Methods Observational study of patients with severe COPD performing daily
spirometry via a telemonitoring system (Medcare Systems, Australia). Measurements were performed, over a 4-week exacerbation-free period. Patient
characteristics (mean SD): n = 14, M/F: 6/8, age 69 11 yrs, FEV1
0.82 0.33 L, previous PC familiarity 64%. Initial supervised training was
provided to all patients. Spirometer calibration was checked at the beginning
and end of the study.
Results Results are presented as coefficients of variation (CoV).
CoV (%) (n = 14 subjects)
FEV1
FEV6
FVC
IC
PEF
RR
SpO2
Mean (SD) 7.0 (2.4) 6.6 (2.3) 7.5 (3.0) 7.6 (2.7) 12.4 (6.2) 10.1 (3.4) 1.7 (1.0)
Range
3.912.7 3.812.0 1.814.0 4.312.0 6.631.1 6.519.8 0.74.0
No calibration errors were detected.

Discussion The reproducibility of these data compares favourably with published data for laboratory-based measurements and provides reference data
for future studies involving serial monitoring in patients with COPD.
Conclusion Patients with severe COPD are able to perform home-based
spirometry with reproducibility similar to that achieved in laboratory
measurements.
TP 110
TOLERABILITY OF MUSCLE MICROBIOPSY DURING COPD

EXACERBATIONS IN THE MELBOURNE LONGITUDINAL COPD
COHORT (MLCC)
M HANSEN1, M THOMPSON1,2, D SMALLWOOD2, GP ANDERSON1,
L IRVING2
1
Pharmacology Department, CRC for Inflammatory Diseases, University of
Melbourne, Victoria 3050, and 2Dept. Respiratory Medicine, Clinical
Epidemiology and Infectious Diseases, Royal Melbourne Hospital, Parkville,
3050
The technique of skeletal muscle microbiopsy has previously been validated [1]
and shown to be minimally invasive and well tolerated in participants with
stable COPD.
Aim A study was undertaken to determine the feasibility and tolerability of
obtaining microbiopsy muscle samples from the patient admitted for acute
exacerbation of COPD patient.
Methods Written informed consent was obtained to collect the muscle, blood
and sputum samples for research purposes.
Local anaesthetic was injected prior to the insertion of a 16 gauge Bard Max
Core Disposable Biopsy Instrument through the associated guide needle.
Multiple passes (up to 6) were obtained. The patient was asked to evaluate the
experience by rating it on the modified Borg scale 010.
Results To date 5 patients and 3 controls have participated in this study. The
GOLD severity ranged from 24 and ATS exacerbation severity 23. The mean
age 75 years (range 6883 years), BMI mean 23.6 kg m-2 (range 17.2
27.1 kg m-2) and fat free mass was determined using single frequency bioimpedance. The sample mass obtained ranged from 27.2104.1 mg, with an
increasing yield occurring with increased experience of the operator.
The procedure has been well tolerated, the Borg scale rating ranged from
12/10. All patients were ambulant post procedure; no haematoma or bruising
was observed in any of the subjects.
Conclusion The microbiopsy technique allows the collection of muscle
tissue with minimal discomfort to the participant. Small tissue masses such as
these are sufficient to obtain measures of local markers of wasting and may
prove to be a useful adjunct to the collection of sputum and blood for the
measure of biomarkers in COPD research.
Reference
1. Hayot M, Michaud A, Koechlin C, Caron MA, LeBlanc P, Prefaut C, Maltais
F. Eur Resp J. 2005;25:431440.
TP 112
TP 111
A PROBLEM BASED ASSESSMENT OF ASTHMA AND COPD IN

AN OLDER POPULATION
VENTILATION HETEROGENEITY OF THE ACINAR AIRWAYS IS

RELATED TO VENTILATION HETEROGENEITY MEASURED
USING SPECT/CT IN COPD
VANESSA M MCDONALD1,3, JODIE L SIMPSON1,3, ISABEL HIGGINS2,3,

PETER G GIBSON 1,3
1
Respiratory & Sleep Medicine, HMRI, 2CPOD, John Hunter Hospital,
NHMRC Centre for Respiratory & Sleep Medicine, and 3The University of
Newcastle, Newcastle NSW 2300
CHANTALE DIBA1,2,5, CHERYL SALOME1,2,5, NORBERT BEREND1,2,

BEN HARRIS1,3,5, DALE BAILEY4,5,GREG KING1,2,3
1
Woolcock Institute of Medical Research, NSW 2050, 2CRC for Asthma,
NSW 2050, 3Department of Respiratory medicine Royal North Shore
Hospital, NSW 2065, 4Department of Nuclear Medicine Royal North Shore
Hospital, NSW 2065, and 5The University of Sydney, NSW 2050
Introduction Older people (OP) with obstructive airways disease (OAD)

experience multiple problems that may impact on their quality of life (QOL) and
disease management. These problems may relate to pathophysiology, symptoms, self management skills, psychological issues, lifestyle or other problems identified as important by the patient.
Aim The aim of this study was to determine the frequency of clinical problems
associated with OAD and to determine if a problem based assessment (PBA)
could adequately identify these problems.
Methods A multidimensional assessment tool was developed and the
content compared to Clinical Practice Guidelines. Participants over 55 years
with diagnosed OAD underwent this assessment.
Results Sixty-one consecutive patients, aged 5987 years, with mean (SD)
FEV1 of 51.4 (17.85) % predicted were assessed. The assessment tool identified a mean (SD) of 3.03 (2.13) current and significant co morbidities with an
additional 11 (3.37) clinical problems per patient. QOL was increasingly
impaired with an increasing number of problems (p < 0.0001). Regression
modelling identified that the number of identified clinical problems accounted
for 55% of the QOL impairment. The model demonstrated that every additional
patient problem was associated with a clinically significant change in QOL
impairment (4.22 units).
Conclusions OP with OAD experience multiple clinical problems and co
morbidities that adversely impact their QOL. A PBA of OP with OAD identifies
significant problems that may not be addressed in a diagnosis centred
approach. There is a need to identify and effectively manage this array of
problems in clinical practice.
Background In COPD ventilation heterogeneity, as measured by the multiple

breath nitrogen washout technique (MBNW), is increased in both the acinar
airways (Sacin) and the conducting airways (Scond) (Verbanck AJRCCM
1998).
Aim To determine if Sacin or Scond, measured by the MBNW correlates with
ventilation heterogeneity measured using single photon emission computed
tomography combined with computed tomography (SPECT/CT) imaging.
Methods Three subjects with moderate to severe COPD had measurements
of ventilation heterogeneity by MBNW and by SPECT/CT imaging. Ventilation
heterogeneity from MBNW was quantified by Sacin, for the acinar airways, and
Scond, for the conducting airways. Ventilation heterogeneity from the
SPECT/CT was quantified by the coefficient of variation (COV) of the voxel
frequency distribution from images.
Results Mean 95%ci age, % predicted FEV1, Sacin, Scond and COV
were 76.3 5.8 years, 42.0 7.1 %, 1.05 0.38, 0.02 0.04 and
5.59 5.42 respectively. There was a trend towards a relationship between
COV and Sacin (r = 0.95, p = 0.19) but not Scond (r = 0.03, p = 0.98).
Conclusions These preliminary data suggest that ventilation heterogeneity
of the acinar airways measured using MBNW can be detected by SPECT/CT.
However, since Sacin is likely to be determined by peripheral lung units that are
below SPECT/CT resolution, these findings suggest that Sacin in COPD is
influenced by ventilation heterogeneity in airways proximal to the acinar
airways.
Funded by CRC for Asthma and Boehringer Ingelheim.
Nominations for awards None.
TP 113
CHRONIC OBSTRUCTIVE PULMONARY DISEASE PHENOTYPES

CAN BE IDENTIFIED BY FEV1 AND CARBON MONOXIDE
TRANSFER FACTOR CORRELATION
JENNIFER PERRET1,2, PETER ROCHFORD1,2, CHRISTINE MCDONALD1,2,
ROBERT PIERCE1,2
1
Department of Respiratory and Sleep Medicine, Austin Hospital, and
2
Institute for Breathing and Sleep, Heidelberg, Victoria 3084
Amongst patients with COPD, those with predominant airways disease (AD)
may have a selective impairment of FEV1, and those with predominant emphysema (E) may have a selective impairment of gas exchange. We looked in
cross-section at relation-ships between FEV1 and CO transfer factor with and
without correction for effective alveolar volume (Kco, DLco) to see if groups
reflecting these phenotypes could be identified.
Methods 200 patients with COPD underwent spirometry and CO diffusing
capacity tests over an 18 month period, and data were analysed using linear
regression with a non-parametric smoother applied.
Results The mean FEV1 was 61 (19.5 SD) % predicted, DLco 55.6 (17 SD)
% predicted, Kco 62.5 (16 SD) % predicted and pack year history 51.8 (30.4
SD). Figures show the relationship of Kco/DLco with FEV1:
Co-efficient
with FEV1
Kco (95% CI, p-value)

0.104 (-0.010, 0.219; 0.075)
DLco (95% CI, p-value)

0.416 (0.309, 0.524; <0.001)
A53
TP 114
VARIABILITY OF FEV1 IN CLINICALLY STABLE NON-CF

BRONCHIECTASIS PATIENTS
ANNA TAI1, TAM EATON1, CHRIS LEWIS1, PAM YOUNG1,
WENDY FERGUSSON1, JOHN KOLBE1,2
1
Greenlane Respiratory Services, Auckland City Hospital, and 2Department
of Medicine, University of Auckland, Auckland
FEV1 is frequently used as an outcome measure in bronchiectasis studies. The
within-day variability of FEV1 in clinically stable non-CF bronchiectasis has not
been studied previously.
Aims (i) To determine the within-day and between-days variability of FEV1 in
clinically stable non-CF bronchiectasis patients. (ii) To determine the effect of
airways clearance on FEV1.
Methods FEV1 was measured pre and post standardized supervised chest
physiotherapy on 3 days over a week in clinically stable non-CF bronchiectasis
patients.
Results 36 clinically stable non-CF bronchiectasis patients were recruited
with baseline characteristics as follows: mean age 63 years (range: 3383);
gender female 67%; mean FEV1 (% predicted): 57.8% (range 24%106%).
The mean inter-day absolute change in pre-chest clearance FEV1 between
three days was 0.009 L (95% CI -0.19 to 0.19 L). The mean change in FEV1
(% predicted) was 0.23% (95% CO -7.4% to 8.2%). Airway clearance does not
significantly change FEV1 with a mean intrapatient difference pre and post
chest physiotherapy FEV1 of 0.003 L (-0.19 L to 0.19 L).The Coefficient of
Variation in between-day FEV1 was 5% reflecting minimal intertest variability in
FEV1. The Intraclass Correlation Coefficient for between-day FEV1 measurements was 0.9.
Conclusion Between-day FEV1 is highly repeatable in clinically stable
non-CF bronchiectasis patients. Chest clearance does not change FEV1 significantly. A change in FEV1 190 ml or a change in % predicted FEV1 of >
8% cannot be attributable to intertest variability alone and should be regarded
as potentially of clinical signficance. This information is an important prerequisite in the interpretation of longitudinal changes in lung function in this patient
group.
Discussion In this diverse group of COPD patients, there was a positive

correlation between DLco and FEV1, but not Kco and FEV1. The FEV1/ Kco
plot identifies substantial numbers of patients with the potential AD and E
phenotypes defined above. We intend to study inflammatory biomarkers in
these groups.
TP 115
MEASUREMENT OF TOTAL BODY WATER AND FAT FREE MASS

IN COPD: DIFFERENT METHODS DO NOT GIVE THE SAME
RESULT
ALISTAIR MILLER1, BOYD J STRAUSS2, ANDREW KYOONG1, PAUL FINLAY1,
PAUL GUY1, STIJN MOL3, PETER HOLMES1, PHIL BARDIN1
1
Department of Respiratory and Sleep Medicine, Monash Medical Centre
Melbourne, Victoria 3168, 2Department of Medicine, Monash University,
Melbourne, Victoria 3168, and 3Department of Respiratory Medicine,
University of Maastricht, The Netherlands
Fat free mass index (FFMI) is a marker of morbidity and mortality in COPD.
Measurement of FFM in the out-patient population is commonly undertaken
using Single Frequency Bioelectrical Impedance Analysis (BIA). However the
formulae to convert measured values to FFM are population dependent.
Schols et al (Am J Clin Nutr, 1991) suggested that formula used for the general
population may be inappropriate for patients with COPD, and derived a specific
formula from total body water (TBW) as measured by deuterium dilution. We
compare this method of measuring FFM with 5 others, along with TBW and
FFM hydration.
Methods TBW was measured in 31 outpatients with COPD by BIA and a
Difference Method (weight-(protein+bone mineral+fat+non-bone mineral+ glycogen)) and FFM hydration was calculated. FFMI was measured by skin fold
anthropometry (SFA), BIA (3 separate formulae), Dual Energy X-ray Absorptiometry (DEXA) and Total Body Potassium by g-counter (TBK). Comparison
between methods for TBW and FFMI was made by Bland-Altman analysis and
between methods of calculation of FFM hydration by paired t-test.
Results The two methods of assessment of TBW showed little difference
(bias -0.04, 95% limits of agreement -5.40 to 5.31). However there was a
significant difference in calculation of hydration of FFM (p = 0.0001). SFA, BIA
(Lukaski), BIA (Tanita) and TBK underestimated FFMI when compared to BIA
(Schols), with bias of -1.24, -3.87, -1.06 and -2.76 respectively. DEXA
however had a bias of only 0.05 and 95% LOA of -3.09 to 3.21.
Conclusions There are differences between methods of assessment of
TBW and FFMI and comparing values between methods must be done with
caution. This has implications for assessment of morbidity and mortality in
COPD.
TP 116
COPD OBSERVATION AND SURVEILLANCE TRIAL (COAST

STUDY): A PILOT STUDY OF COPD EXACERBATIONS
AMANUEL TESFAI, PETER HOLMES, ISABEL HAYES, JOANNE MCKENZIE,
HAYAT DAGHER, PHILIP BARDIN
Department of Respiratory and Sleep Medicine, Monash Medical Centre,
Clayton, Melbourne
Chronic obstructive pulmonary disease (COPD) has been identified as a major
health problem in Australia. Recent studies have suggested that respiratory
viral infections are the major cause of a worsening of COPD; however this has
not been studied in Australia.
Aim To characterize PEF changes and identify viruses during COPD
exacerbations.
Methods A pilot prospective longitudinal cohort study was done. Patients had
confirmed COPD with FEV1 <70% predicted and reversibility <10% and/or
200 ml. Patients recorded daily peak expiratory flow (PEF) measurements and
daily chest and cold scores over a period of 2 years. Sputum samples and
nasal aspirates were taken at 6-month review (control visit) and whenever they
had symptoms of an exacerbation (worsening of COPD symptoms Seemungal et. al. Am J Resp Crit Care Med, 2001). Nasal aspirates and sputum
samples were obtained and analysed by RT-PCR for rhinovirus (RV).
Result Five patients have finished 2 years of study. A total of 12 exacerbations were reported based on patient symptoms. Only 3 exacerbations were
associated with significant reductions in PEF and only one was linked to
increases in nasal cold scores. All samples taken at control visits and nasal
aspirates and sputum samples during exacerbations were negative for RV by
RT-PCR. Positive controls confirmed the accuracy of the assay.
Conclusion Our data suggest that a symptom-based definition of COPD
exacerbation is not always accompanied by significant reductions in lung
function parameters. These exacerbations are also not associated with the
commonest reported viral cause. Our findings suggest that variability of COPD
may mimic.
A54
TP 117
EFFECT OF INCREASING DOSES OF MANNITOL ON MUCUS

CLEARANCE IN PATIENTS WITH BRONCHIECTASIS
EVANGELIA DAVISKAS1, SANDRA D ANDERSON1, STEFAN EBERL2,
IVEN H YOUNG1
1
Departments of Respiratory & Sleep and 2PET & Nuclear Medicine Royal
Prince Alfred Hospital, Camperdown, NSW 2050, Australia
Bronchiectasis is characterized by hypersecretion of mucus and impaired
clearance that results in mucus accumulation, chronic cough, sputum production and recurrent infections. Inhaled mannitol (400 mg) improves clearance of
mucus by increasing the airway hydration and by reducing the viscoelastic and
surface properties of mucus. However, the effect of other doses of mannitol on
the clearance of mucus in patients with bronchiectasis is unknown.
Methods Fourteen patients, age: 63.3 5.7 yr, were studied on 5 visits.
Clearance of mucus was measured using 99mTc-sulphur colloid and imaging
with a gamma camera at baseline and with mannitol (160, 320 and 480 mg),
over 45 minutes and over a further 30 minutes involving 100 voluntary coughs.
A control study assessed the effect of cough provoked by mannitol during the
intervention.
Results Whole right lung clearance over 45 minutes was 4.7 1.2% and
10.6 2.6% on baseline and control days and increased to 16.7 4.2%,
22.8 4.2% and 31 4.7% with mannitol 160, 320 and 480 mg respectively
(p < 0.001). Clearance over 45 min with 480 mg mannitol was greater than
clearance with 320 (p < 0.02) and 160 mg (p < 0.0001). Total clearance over 75
min, after mannitol and voluntary coughs, was 36.1 5.5%, 40.9 5.6% and
46.0 5.2% with 160, 320 and 480 mg mannitol respectively, all significantly
different to baseline (24.1 6.0%) (p < 0.006) and control (13.1 3.0%)
(p < 0.0001). Total clearance over 75 min with 480 mg mannitol was greater
compared to 160 mg (p < 0.03).
Conclusions Mucus clearance increases with increasing doses of mannitol
and it can be further increased by cough in patients with bronchiectasis.
Supported by the NHMRC
Nomination Nil.
Conflict of Interest Yes.
TP 118
CIGARETTE SMOKE-INDUCED LUNG INFLAMMATION AND

SKELETAL MUSCLE WASTING ARE NOT ALTERED BY A
HIGH-FAT DIET IN MICE
MICHELLE HANSEN1, HUI CHEN3, JESSICA JONES1,
SHENNA LANGENBACH1, ROSA GUALANO1, ROSS VLAHOS1,2,
MARGARET MORRIS3, GARY ANDERSON1,2
CRC for Chronic Inflammatory Diseases and Departments of 1Pharmacology
& 2Medicine, University of Melbourne, Vic 3010, and 3Department of
Pharmacology, University of NSW, NSW 2052
Weight loss and skeletal muscle atrophy are major determinants of morbidity in
Chronic Obstructive Pulmonary disease (COPD), which are independent of
lung function impairment. Thus, we examined if a high-fat diet (HFD) protected
against the wasting associated with prolonged cigarette smoke exposure (SE)
in mice.
Methods Male BALB/c mice were exposed to the smoke of 4 cigarettes/day,
6 days/week for 7 weeks. Sham mice were handled identically without smoke
exposure. Mice consumed either standard laboratory chow (3.5 kcal/g, consisting of 12 % fat) or a HFD (4.3 kcal/g, 32% consisting of fat). We examined
the effect of SE and HFD on hind limb skeletal muscles, lung (tissue &
bronchoalveolar lavage (BALF)) and systemic inflammation in the 4 groups of
mice (n = 8/ group).
Results After 7 weeks of HFD, sham and SE mice were 12 and 13% heavier
(respectively, P < 0.05) than chow fed animals. Conversely, SE significantly
decreased body weight of chow and HFD fed mice by 16 and 15%, respectively, compared to sham animals (P < 0.05). The HFD did not protect against
the decrease in soleus, tibialis anterior and gastrocnemius skeletal muscle
weights induced by SE (P < 0.05). SE altered the mRNA expression of a
number of genes associated with the regulation of skeletal muscle mass
including insulin-like growth factor-I (IGF-I), atrogin-1 and interleukin (IL)-6. The
mRNA expression of pro-inflammatory cytokines and chemokines was significantly increased by SE in the lung, as were the number of inflammatory cells
in BALF (P < 0.05). On the other hand, although obesity has been linked to
systemic inflammation, the HFD exerted little direct effect on the skeletal
muscle and lung parameters measured. SE and HFD had no effect on two
markers of systemic inflammation, IL-6 and serum amyloid A, whereas SE
tended to reduce circulating IGF-I, an anabolic hormone.
Conclusions The HFD was not protective against the weight loss and
skeletal muscle wasting associated with cigarette smoke exposure.
Supported by the NHMRC and CRC for Chronic Inflammatory Diseases.
Cystic Fibrosis SIG: Poster Session

TP 120
TP 119
THE IMPACT OF PHYSIOTHERAPY INTERVENTIONS ON

GASTRO-OESOPHAGEAL FUNCTION IN COPD AND
BRONCHIECTASIS
A LEE1, L DENEHY1, S ROBERTS3,5, R STIRLING4,5, J WILSON4,5,
B BUTTON1,2
1
School of Physiotherapy, The University of Melbourne, Vic 3010,
Departments of 2Physiotherapy, 3Gastroenterology and 4AIRMED, The
Alfred, Vic 3004, and 5Department of Medicine, Monash University, Vic 3800
Background Patients with COPD and bronchiectasis undertake airway
clearance therapy (ACT) and exercise as part of physiotherapy management
but it is unknown whether these treatments provoke gastro-oesophageal reflux
(GOR). This study aimed to determine the impact of positive expiratory pressure (PEP) therapy and exercise on gastro-oesophageal function.
Methods During dual-probe 24 hour oesophageal pH monitoring, all patients
undertook a single session of PEP therapy, measures of exercise capacity
(six-minute walk distance [6MWD]) and upper limb function (Grocery Shelving
Task [GST]). The number of reflux episodes (NRE) and % reflux time (RI) were
recorded during each intervention and compared to equivalent background
time (BGT).
Results Fifty-seven patients (30 with bronchiectasis, 27 with COPD
[mean SD FEV1 61% predicted 24.6]) completed all interventions. Episodes of distal oesophageal reflux occurred in 31% of patients during PEP
therapy, 24% during 6MWD and 25% during GST. However, there was no
significant difference in RI during 6MWD or PEP therapy compared to BGT (all
p > 0.05). NRE were decreased compared to BGT during the GST (p = 0.001)
and 6MWD (p = 0.001). Those diagnosed with GORD had higher RI during
GST (p = 0.005) and 6MWD (p = 0.04) compared to those without GORD.
Conclusions Walking and upper limb function provokes distal GOR in
patients diagnosed with GORD, but the extent of GOR was not significant. ACT
using PEP in upright sitting was not associated with increased GOR episodes.
Funding source NHMRC, The University of Melbourne, Monash University,
Physiotherapy Research Foundation.
INFLAMMATION AND INFECTION IN INFANTS WITH CYSTIC

FIBROSIS IS ASSOCIATED WITH CHANGES IN LUNG
FUNCTION IN THE FIRST YEAR OF LIFE
G NOLAN1,2, S BRENNAN2, P SLY1,2, S STICK1,2, S RANGANATHAN3,
B LINNANE3, GL HALL1,2 for the Perth & Melbourne Detection of Early Lung
Disease in CF Program
1
Respiratory Medicine, PMH, 2Clinical Sciences, ICHR, Perth, and 3Thoracic
Medicine, RCH, Melbourne
Introduction Infection and inflammation is assessed with bronchoalveolar
lavage (BAL) in the clinical management of infants with cystic fibrosis (CF) but
is unsuitable for regular use. Lung function is routinely used in older children to
track changing clinical status in CF lung disease. We aimed to ascertain the
role of lung function (ILF) tests in assessing clinical status of infants with CF.
Methods Infants with CF diagnosed by new born screening underwent BAL
and ILF testing at diagnosis and again at 15 months of age. Tests of airway
(Raw) and respiratory tissue mechanics (dampingGrs and elastanceHrs)
using low frequency forced oscillations (LFOT) & ventilation distribution (Lung
clearance index-LCI and moment ratios-MDN) from multiple breath washouts
(MBW) were performed under sedation 12 days prior to BAL. Associations
between ILF variables and inflammation (IL8, total cell count (TCC) and neutrophil numbers) and infection were tested accounting for repeated testing and
body length.
Results Tests of ILF were obtained in 26 infants aged 13.5 weeks at visit 1
and 64 weeks at visit 2 (9 male), resulting in 17 then 22 (LFOT) and 21 then 21
(MBW) measurements at visits 1 and 2 respectively. Inflammatory and infection
status was confirmed in all infants. Peripheral but not airway mechanics worsened with infection (Grs & Hrs) and inflammation (Grs: TCC & neutrophils, Hrs:
IL8), while impairment of MDN but not LCI was associated with TCC and
neutrophils.
Conclusions These data demonstrate that changes in inflammation and
infection impact on peripheral lung function in infants with CF and that serial
testing may be useful adjunct to BAL in their clinical management.
Supported by NHMRC and US CF Foundation.
TP 121
DEVELOPING SALIVARY IMMUNOGLOBULIN A AS A USEFUL

MARKER FOR PSEUDOMONAS AERUGINOSA RESPIRATORY
INFECTION
L GARRATT1, T DOUGLAS1,2, S BRENNAN1, S STICK2,3, P RICHMOND3,
P SLY1,2
1
ICHR, Perth, WA 6008, 2Department of Respiratory Medicine, PMH, Perth,
WA 6008, and 3SPACH, UWAl, Perth, WA 6008
P. aeruginosa is a significant opportunistic lung pathogen in individuals with
cystic fibrosis (CF) and is associated with increased lung disease and morbidity. Early intervention is beneficial for the effective clearance of P. aeruginosa
and better long-term health outcomes. Currently, lung flora of CF patients is
monitored by regular culturing of sputum, however, children unable to expectorate are limited to annual bronchoalveolar lavages (BAL), which is invasive
and requires general anaesthesia. Saliva is useful for clinical assays as collection is simple, non-invasive. We are developing a standardized enzymelinked immunosorbent assay (ELISA) to detect respiratory infection of P.
aeruginosa in CF children who cannot expectorate.
Methods 18 children (718 years) with CF and recent P. aeruginosa lung
infection history and 16 non CF children (16 years) with no previous P.
aeruginosa infection history provided saliva as positive, negative controls
respectively. Saliva was obtained by spitting, or absorbed using cellulose
swabs and later extracted. These cell-free supernatant samples were used in
an ELISA anti-P. aeruginosa IgA using commercial antigen. All results were
standardized to account for flow using total IgA expression.
Results Median value was increased 9 fold in the recent P. aeruginosa lung
infection group (Mann-Whitney test, n = 34, p 0.001). There was no significance between mucoid and non mucoid samples, and detection was independent of cfu/ml.
Discussion Early findings support that P. aeruginosa respiratory infection
can be detected through specific analysis of salivary IgA expression. Larger
population sampling (30 positive, 90 negative) will aid selection of cut-off
values for specificity and sensitivity testing in the future to objectively determine
the utility of this assay as a means of monitoring for P. aeruginosa and for
determining effectiveness of treatment.
A55
TP 122
AIRWAY SURFACES IN MICE: SYNCHROTRON XRAY IMAGING

OF MCC, ULTRA-HIGH RESOLUTION CT, AND IMPROVED EDGE
DETECTION WITH MICROBEADS
DS PARSON1, E WONG1, RR BOUCHE2, K UESUGI3, N YAGI3, KKW SIU4
Womens & Childrens Hospital, & Dept of Paediatrics, University of
Adelaide, SA, 2University of Nth Carolina at Chapel Hill, US, 3Spring-8
JASRI, Japan, and 4School of Physics & Monash Centre for Synchrotron
Science, Monash University,Vic
Current non-invasive imaging methods for lung (e.g. HRCT, MRI, PET) are
valuable for detecting and monitoring gross change in human CF airways, but
resolution is limited e.g. HRCT detects airways > 1.5 mm dia. Detection of
airway structures in rodent airways demands significantly greater resolution
and rapid image capture. We have developed 2-D and 3-D methods able to
visualize the smallest mouse airways non-invasively using synchrotron phase
contrast Xray imaging (PCXI). Methods: Anaesthetized mice were imaged at
the SPring-8 synchrotron (Japan). Images were captured on CCD detectors
(1.1 mm or 0.45 mm square pixel arrays, 100 cm propagation distance, 25 keV,
repeated 100300 ms exposures for up to 45 mins. 3-D CT data (voxels
12 12 12 mm3) was obtained in post-mortem mice. Volume renderings were
produced using Volview or OsiriX software. Results: Live, 2-D nasal or tracheal
imaging captured airway-surface activity consistent with mucociliary clearance.
Resolution was ~2 mm. Instilled glass beads (<2 mm dia) imporved airwaysurface contrast. Mouse lung CT slices, and dynamic (fly through) 2D and 3D
sequences visualized airway branching to approx ~100 mm dia. Discussion:
Synchrotron PCXI provides new options for non-invasive imaging, able to
resolve small airways in mice. The higher airway-edge contrast when using
glass beads may assist detection of airway surface liquid. Volume reconstructions provide new option studies of structure-function relationships with airway
disease. The potential of this non-invasive method is yet to be realized; continued improvements in detector & digital technology should advance the
achieveable resolution and the modes of visualization.
Support NHMRC, USA CFF, philanthropic donations. DP, KS supported in
part by the AMRF Program.
TP 124
PSEUDOMONAS SEROLOGY AS A MARKER OF RESPIRATORY

INFECTION IN PRESCHOOL CHILDREN AND INFANTS WITH
CYSTIC FIBROSIS
TP 123
GENE TRANSFER IN MOUSE & SHEEP LUNG WITH A

LENTIVIRUS VECTOR
C LIU1, 2, E WONG1,2,3, CK TAN3, G SMITH1, D ANSON3,4, D PARSONS1,2,4
Department of Pulmonary Medicine, 2Child Health Research Institute,
3
Department of Genetic Medicine, Womens and Childrens Hospital, and
4
Dept of Paediatrics, University of Adelaide, Adelaide, SA
Background We have previously shown robust lentivirus-mediated gene

transfer is possible in mouse nasal airway after pretreatment of L-alysophosphatidylcholine (LPC). We have now tested this technique in mouse
and sheep lung.
Methods Different concentrations (0.03%1.0%) and volumes (15 mL30 mL)
of LPC were administered to the trachea of C57L/B6 mice trans-orally. In sheep
4.5 ml of 0.3%0.1% LPC was delivered to the main bronchus of right lung of
sheep via bronchoscope. One hour later 30 ml75 ml (mice), or 0.45 ml (sheep)
of LV-LacZ vector (3.12 109 TU/ml) was delivered in the same manner. After
7 days lungs were inflation-fixed in situ, stained using X-gal, sectioned and
stained with Saf-O or H/E.
Results Mice treated with low doses (concentration & volume) of LPC, and a
low volume of virus, showed scattered LacZ gene expression in the trachea.
When a high concentration (1.0%) or volume (30 ml) of LPC, or a high volume
of vector was used there was extensive gene transfer in trachea, carina &
bronchi in most lobes. In sheep, gene expression extended from branch 5 of
the right lung bronchus to peripheral small airways. The highest expression
was always near delivery site. Transduced epithelial cells included ciliated cells
and basal cells in both species.
Conclusions Strong gene transfer can be achieved in mouse lung with
appropriate doses of LPC and vector, and the data suggest dose-dependency
of gene expression for both LPC and the vector. Although modest in these
initial studies, gene transfer was also achieved using the lentivirus vector with
and LPC pre-treatment in sheep lung airways. We are continuing to seek
further improvement in efficiency of gene expression in lung airways.
Supported by NH&MRC, USA CFF, Ch7/Novita CRF, Philanthropic donations.
L BERRY1, S BRENNAN1, PD SLY1,4, C WAINRIGHT2, K GRIMWOOD3

ACFBAL study group, TA DOUGLAS1,2
1
Institute for Child Health Research, Perth, Western Australia 6008, 2Royal
Childrens Hospital, Herston, Queensland 4029, and 3Department of
Respiratory Medicine, Princess Margaret Hospital for Children, Perth,
Western Australia 6008
Background Effective eradication of Pseudomonas aeruginosa (Psa) in
young children is dependent on accurate and early detection of infection. In
preschool children detection of Psa may involve invasive or insensitive
methods such as bronchoalveolar lavage (BAL) and oropharyngeal (OP)
swabs. The value of Psa serology as potential markers of early respiratory
infection with Psa remains uncertain in very young children.
Aims To determine the value of Psa serology in young children with CF as
markers of early/initial Psa infection diagnosed by BAL and OP culture.
Methods A commercially available ELISA was used to determine serum IgG
levels to multiple pseudomonas antigens. Paired serum and BALF from a
discovery population of 76 preschool children and infants with CF in Perth, WA
was analysed to determine optimal serology cut off levels using a positive
culture of Psa BALF as gold standard. Sera from a test population of preschool
children recruited to the Australasian CFBAL Study were then assayed using
the ELISA and applied cut-off values in a blinded fashion. ROC curves were
generated to calculate the positive and negative predictive values of serology
in predicting both positive culture of Psa in OP swab and in BAL (paired
samples).
Results Paired serum/BAL (n = 74) and serum/OP swab (n = 28) from 83
children 0.15.5 years (mean 1.94) in the test population were analysed.
Positive serology poorly predicted the presence of Psa in BALF culture
(PPV = 39%) whilst negative serology produced a higher NPV of 86%. In
contrast positive serology better predicted the presence of Psa in upper airway
culture (PPV = 71%, NPV = 84%).
Conclusion Positive serology may be a better marker of upper airway
infection/colonization with Psa than lower respiratory tract infection in young
children. The value of serology specifically as a marker of lower respiratory
infection appears greatest in excluding Psa infection.
Funded by Australian CF Research Trust, USCF Foundation and NHMRC.
A56
TP 125
LUNG FUNCTION USING FORCED OSCILLATIONS IS WORSE IN

YOUNG CHILDREN WITH CYSTIC FIBROSIS INFECTED WITH
PSEUDOMONAS AERUGINOSA
CL GANGELL1, GL HALL1,2, SM STICK1,2, S BRENNAN3, PD SLY1,2,3
1
School of Paediatrics and Child Health, The University of Western Australia,
2
Respiratory Medicine, Princess Margaret Hospital, and 3Clinical Sciences,
Telethon Institute for Child Health Research
Background In cystic fibrosis (CF) infections with Pseudomonas aeruginosa
in adults is associated with worse lung disease. The aim of this study was to
determine the effect of an infection with P. aeruginosa on lung function in
preschool aged children with CF.
Methods Lung function using the forced oscillation technique (FOT) was
recorded in children with CF the morning of bronchoalveolar lavage (BAL).
Nine children with CF who never had a detectable infection at BAL formed the
control group. Lung function was measured in 11 children infected with P.
aeruginosa. Seven children (of 11) had FOT 3 months after P. aeruginosa
eradication. Respiratory resistance (Rrs) and reactance (Xrs) data at 6 Hz were
obtained and expressed as Z scores. Differences between control and infection
groups were analysed using an independent t-test, and changes in Rrs and Xrs
following eradication analysed using a paired t-test.
Results Children infected with P. aeruginosa had worse lung function compared to CF controls [Z scores: Rrs6: 1.22 1.68 vs. -0.26 0.56; p = 0.02.
Xrs6: -1.16 1.37 vs 0.19 0.66; p = 0.01, respectively]. Lung function had
not returned to normal within 3 months after eradication of P. aeruginosa.
Conclusion Infection with P. aeruginosa is associated with worse lung function using the FOT in young children with CF; however abnormalities do not
improve within 3 months following an eradication protocol.
Support The Australian Cystic Fibrosis Research Trust and US Cystic
Fibrosis Foundation.
TP 126
GENE TRANSFER EFFICIENCY IN PAEDIATRIC CYSTIC

FIBROSIS EPITHELIAL CELL CULTURES
E SUTANTO1, A KICIC1,2, S STICK1,2
Telethon Institute for Child Health Research, Western Australia, and 2School
of Paediatrics and Child Health, The University of Western Australia
1
Background CF is caused by mutations in the CF transmembrane conductance regulator (CFTR) gene that result in improper folding, defective translocation and subsequent loss of function to a variable extent. One of the many
methods to restore CFTR expression and function is via gene transfer. To date,
there have been many studies using viral and non-viral vehicles for transgene
delivery, however little data is available about the use of such vehicles in
paediatric primary epithelial cells and the effect this might have on the cells.
Thus, the aim of this study was to determine and compare gene transfer
efficiency between non-viral and viral delivery in the CF airway epithelial cells
(AECs).
Methods AECs were obtained from CF patients by non-bronchoscopic
brushing. Cells were transfected or transduced with a vehicle carrying green
fluorescent protein (GFP) as a marker. Gene transfer efficiency was determined by direct observation of GFP expression and compared to 16HBE cells
as controls.
Results Adenovirus (Ad) vehicle transduced both nasal and tracheal epithelial cells from children with CF with high efficiency (>95% total cells), with no
observed morphological cytotoxic effect. 16HBE cells however demonstrated
cytotoxicity with approximately 50% cell death following Ad transduction. Transfection of CF cells with plasmid DNA carrying GFP transgene resulted in
moderate transfer efficiency.
Conclusions CF primary cell cultures can be transduced efficiently via both
methods of delivery with no sign of cytotoxicity. This method has provided proof
of principal that the CF primary cells, which are known to be difficult to
transfect/transduce to express transgene effectively. In light of this, restoration
of CFTR expression via one of these vehicles is a useful technique to aid
investigation of CFTR pathophysiological processes in CF epithelial cells.
Funding sources CHRF.
TP 128
LONGER TERM OUTCOME OF LAPARASCOPIC

FUNDOPLICATION IN PATIENTS WITH CF WITH AND WITHOUT
LUNG TRANSPLANT
TP 127
SCREENING FOR CYSTIC FIBROSIS RELATED DIABETES IN

CYSTIC FIBROSIS CLINICS IN AUSTRALIA AND NEW ZEALAND
A SMITH1, P BERGMAN1, D ARMSTRONG2
Diabetes Ambulatory Care Service and 2Department of Respiratory and
Sleep Medicine, Monash Medical Centre, Clayton, VICTORIA 3168
1
Introduction Up to 25% of individuals with Cystic Fibrosis (CF), develop

Cystic Fibrosis Related Diabetes (CFRD). Poor glycaemic control in CFRD is a
major contributor to sub-optimal nutrition and deteriorating lung function in
these patients. There is no universally accepted protocol for screening, diagnosing, nor treating patients with CFRD. The aim of this study was to determine
the CFRD screening protocols of CF clinics in Australia and New Zealand
Methods All CF Units (paediatric and adult) across Australia and New
Zealand were surveyed using a postal questionnaire. Demographic data,
together with the presence and method of CFRD screening was obtained.
Results Fourteen of 17 (82%) surveys were completed. 13/14 (93%) centres
used CFRD screening. Triggers for screening were not standardized, and
varied between age (10 yrs to 18 yrs), reduction in FEV1, or development of
symptoms suggesting CFRD. There was also wide variability in the initial
screening method employed, being one or a combination of random blood
glucose level (BGL), fasting BGL, postprandial BGL, HbA1c or the Oral
Glucose Tolerance Test.
Conclusion Although CFRD screening is widely utilized in Australia and New
Zealand CF centres, the rate of CFRD is probably under estimated, due to lack
of standardization in screening methods.
P BURTON1, M LEE1, S ROBERTS2, A SMITH1, G SNELL3, J WILSON3,

W BROWN1, B BUTTON3
Departmenjt of 1Surgery & 2Gastroenterology, and 3AIRmed, The Alfred
Hospital, Melbourne, Vic 3004
Gastro-oesophageal reflux (GOR) is common in CF before and after lung
transplant (LTx). Laparascopic fundoplication (LapF) is used when conservative therapy fails. We reviewed longer term clinical outcomes of LapF in CF
before & after LTx.
Methods A 5 yr. review of patient records was undertaken. Patients completed an outcome questionnaire at the time of the review.
Results 37 patients underwent Lap.F; 21 transplanted (LTxGroup), mean
age 40 (2167) years, and 16 not transplanted (CF Group), 35 (2249) yrs.
Mean time between LTx & LapF was 654 (414; 1133) days. Complete (Nissen)
LapF was undertaken in 30% of patients and partial (Toupet) in 70%. Average
length of stay: LTxGroup: pre-op.7.81; post-op.7.82 days; CF Group 5.2 & 12.5.
At 6 months post-LapF there was no significant difference in FEV1 compared
to pre-op (both groups). FEV1 remained stable up to 300 days post LapF in the
CF Group, p = 0.65. There was a small, significant decline in FEV1 in the LTx
Group, p = 0.02. BMI fell in both groups in the first 100 days post-LapF,
remaining lower at 300 days post-LapF compared to 100 days pre-op,
p 0.001 in the LTx Group. BMI returned to baseline 300 days post-op in the
CF Group, p = 0.47. Patient satisfaction with LapF, improvement in symptoms
& quality of life was high in both groups.There was 1 failed LapF, 2 oesophageal dilatations, 1 LOS hypotonia.Four deaths on Day 19, 253, 284, 382
post-LapF were not attributed to LapF.
Conclusions In this series of patients there was a small but significant drop
in FEV1 in the LTx Group 300 days after LapF, but not in the CF Group. There
was a significant drop in BMI in both groups after LapF, but the CF Group
returned to pre LapF values at +300 days. Patient satisfaction with the procedure was high. A long term prospective study is warranted.
TP 129
USE OF THE AUSTRALASIAN CYSTIC FIBROSIS DATA

REGISTRY FOR RESEARCH: A QUALITY ANALYSIS OF THE
2003 DATA SET
B MARTIN, P COOPER
Dept of Respiratory Medicine, The Childrens Hospital at Westmead, NSW
2145
Introduction The Australasian Cystic Fibrosis Data Registry (ACFDR) was
instituted in 1996 and contains demographic, health and social data from 1998
for cystic fibrosis patients throughout Australia and New Zealand. Annual
summary reports have been produced but little research has been conducted
using the Registry. We sought to examine the 2003 data set for completeness
and quality to help assess its utility for research.
Methods Deidentified data from the ACFDR for 2003 were obtained from
Cystic Fibrosis Australia. Data fields were examined for completeness and
clear discrepancies in data were identified. Association of genotype with pancreatic insufficiency was examined to assess whether the data support current
knowledge. Analysis was carried out using Microsoft Excel and SPSS Version
15.
Results Data were available for 2374 subjects across 114 fields. Completeness of data ranged from 57.3% for fracture as a complication to 100% for
gender. Data were available for height in 78.8% of subjects, weight in 79.1%,
age in 81.6%, genotype in 86.4% and pancreatic status in 95.1%. FEV1 was
available for 65.0% of total subjects and 95.6% of subjects with age 5 years
recorded. Clear discrepancies between related fields, likely due to data entry
errors, were noted in < 1% of entries. The data supported the known association of genotype with pancreatic insufficiency (94.9% in DF508 homozygotes
vs 75.2% in DF508 heterozygotes vs 77.0% in non-DF508 heterozygotes;
p < 0.001).
Conclusion Completeness of data entry within the ACFDR varies across
fields which may have implications for research utility. Most important data
fields, however, are relatively well recorded with a low rate of clear discrepancies. Furthermore, the data support the expected association of genotype with
pancreatic insufficiency which implies adequate quality for further analysis.
A57
TP 130
NITROGEN WASHOUT LUNG CLEARANCE INDEX (LCI) IN

CYSTIC FIBROSIS
J MORTON, B ODONOVAN, B MASON
Department of Respiratory Medicine, Sydney Childrens Hospital, High
Street, Randwick, NSW 2031
In recent times many children with Cystic Fibrosis have minimal lung disease
with normal spirometry. More sensitive tests of peripheral airway function such
as Lung Clearance Index (LCI) measured by nitrogen washout may be clinically
helpful.
Aims To examine the relationship between LCI and spirometric measurements of airway function in mild CF lung disease.
Subjects Were 54 children with CF. 25 females mean age 10.2 yrs (524 yrs)
mean FEV1 87.2% (38134%) mean MMEFR 72% (30120%) and were
clinically well.
All patients performed spirometry and LCI measurements in one study session.
Results LCI measurements were reproducible (5%) within individuals.
39/54 patients had normal spirometry (80134% P) and 15 patients had
abnormal spirometry FEV1 (3880% P)
LCI varied from 613.4. The LCI correlated with both FEV1 and MMEFR
(r2 0.38 for both). Of the 39 patients with normal spirometry
17 had LCI within the normal range (mean 6.8) and 22 had LCI which was
abnormal (mean 9.4).
Summary LCI measurements identified abnormality in 55% of CF patients
with normal spirometry. This test may be more sensitive to early changes in
small airway function in CF.
TP 132
Lung Cancer/Bronchology SIGs:

Combined Poster Session
TREAMENT OF PRIMARY AND SECONDARY PNEUMOTHORAX

UTILIZING MEDICAL THORACOSCOPY
TP 131
AN AUDIT OF THE TRACHESTOMY TEAM SERVICES AT

DANDENONG HOSPITAL
P WONG1, S JUKES2, J BEST2, C ELLIS2, D MANSFIELD2
Department of Respiratory and Sleep Medicine, Royal Melbourne Hospital,
and 2Department of General Medicine, Dandenong Hospital
Background Care of the ward based tracheostomized patient is associated

with a high acute complication rate that may be reduced by care delivered from
a multidisciplinary team.
Aim To assess the impact on complications by specialist multidisciplinary
team approach in the care of patients with tracheostomy in the ward.
Methods A Retrospective audit of ward based tracheostomy care between
20034 was compared to a prospective evaluation of the impact of a specialist
multidisciplinary team (20056). The team comprised of a speech pathologist,
physiotherapist, ICU nurse liason/educator and was overseen by a respiratory
intensivist. The focus of care was around humidification/suctioning, swallowing
assessment and decannulation decision making. Primary outcomes were rates
of sputum plugging with documented consequence, rate of MET (Medical
Emergency Team) or code blue calls; and readmission to ICU. Secondary
outcomes include length of tracheostomy cannulation on the ward.
Results 25 retrospective and 37 prospective ward based tracheostomized
patients were included in the analysis. Major indications for tracheostomy
insertion were post oropharynx surgery, protracted ventilator wean, aspiration
risk and sputum toileting.Type of tracheostomy inserted included surgical,
percutaneous, permanent and mini-tracheostomy. Mean LOS in hospital was
34.6 3.3 days vs 29.1 3.3 (NS). Mean tracheostomy cannulation duration
on the ward was 19.1 3.1 days vs 18.9 3.5 (NS). Ward based complications included sputum plugging 5 (20%) vs 1 (3%), p = 0.01; MET calls 7 (28%)
vs 5(13%), p = 0.07; Code blue calls 4(16%) vs 0 (0%), p < 0.01; ICU readmissions 9(36%) vs 2 (5%), p < 0.01.
Discussion Multidisciplinary tracheostomy care teams can produce striking
reductions in adverse events when initial adverse event rate was high. Length
of stay was not improved in this analysis.
S VINCENT, G SIMPSON
Department of Thoracic Medicine, Cairns Base Hospital, Cairns, QLD 4870
Medical thoracoscopy is utilized widely throughout Europe and Northern
America by thoracic physicians for the management of pleural disease, including the undiagnosed pleural effusion, malignant effusions and less commonly
pneumothorax (PTX). Australia has limited experience in this modality. We
report the success of medical thoracoscopy in both primary and secondary
PTX requiring intervention.
Methods Data were collected from 2001 to 2007 in patients treated with
medical thoracoscopy for the treatment of PTX.
Results 11 patients, 7 male, 4 female. Average age 48 (range 1986). 1 first
episode primary spontaneous (PS) PTX, 2 third episodes of PS, 5 first secondary spontaneous (SS), 1 second SSPTX, 2 third SSPTX. Underlying pulmonary disease in secondary PTX included: 4 chronic obstructive pulmonary
disease, 1 lymphangioleiomyomatosis, 1 mesothelioma, 1 metastatic angiosarcoma and 1 was secondary to a motor vehicle accident. 7 had a history of
smoking, 5 were former smokers and 2 were current smokers, with a mean 24
pack year history (range 545). 7 PTX were large, 4 moderate. 5 patients had
an intercostal catheter (ICC) inserted prior to thoracoscopy, 1 had failed pleural
aspirate. There was evidence of bronchopleural fistula in 7 patients prior to the
procedure. There was a median of 9 days from PTX to thoracoscopy. Light
sedation was used for the procedure in 10 patients, 1 required a general
anaesthesia with a double lumen endotracheal tube due to anxiety. Single port
entry, dry Talc poudrage and a 16 gauge French ICC was used for all procedures. ICC was removed a mean of 2 days following thoracoscopy and patients
discharged on day 4. Pain was the most common complication, requiring
narcotic analgesia. One patient died on day 7, secondary to metastatic
angiosarcoma. There has been no recurrence of PTX in any patient.
Conclusion Medical thoracoscopy, performed by thoracic physicians is an
effective procedure for the treatment of pneumothorax requiring intervention,
including selected patients with evidence of bronchopleural fistula.
Funding Nil.
Nomination for young investigator award No.
A58
TP 133
ANATOMICAL OPTICAL COHERENCE TOMOGRAPHY

PROVIDES ACCURATE CALIBRE MEASUREMENTS IN
PHANTOM AIRWAYS
TP 134
ABSTRACT WITHDRAWN
J WILLIAMSON1,3, J ARMSTRONG2, S BECKER2, D SAMPSON2, J

KIRKNESS1,2, J WALSH3, M PHILLIPS4, D HILLMAN3, P EASTWOOD1,3
1
School of Anatomy & Human Biol, 2Optical and Biomedical Engineering
Laboratory, Univ of Western Australia, 3West Australian Sleep Disorders
Research Institute, and 4Dept Resp Medicine, Sir Charles Gairdner Hospital,
Australia
Background Measurements of regional airway size and function are difficult
to quantify in-vivo with existing bronchoscopic techniques. Anatomical Optical
Coherence Tomography (aOCT) is an imaging technique offering the ability to
provide quantitative calibre measurements of hollow cavities such as the
bronchial tree. We describe the validation of this technique in phantom airway
sections constructed from 9 aluminium circular tubes and 3 plastic rectangular
tubes.
Methods aOCT measurements were performed by passing a catheter (outer
diameter 2.24 mm) containing a rotating fibre-optic probe into each tube.
Lumen area (LA) and diameter was measured using ImageJ software and
compared to measurements obtained with digital callipers. Measurements of
inter- and intra-observer variability were also obtained. To simulate the in-vivo
conditions imposed on the probe, the catheter was also inserted through a
bronchoscope into the airways of a resuscitation mannequin.
Results Compared to measurements obtained by digital callipers (range
452 mm), values obtained by aOCT showed no significant difference in mean
diameter (-0.05 0.11 mm) or LA (-1.75 5.18 mm2). There was minimal
inter- and intra-observer variability [inter-observer mean diameter difference
0.001 0.08 mm, coefficient of variation CV = 0.1%; intra-observer mean
diameter difference -0.028 0.072 mm, CV = 0.2%]. Measurements performed through the mannequin did not differ significantly.
Conclusions In phantom airways of varying size, aOCT provides accurate
diameter and LA measurements, has minimal inter- and intra-observer variability and is not affected by simulating in-vivo bronchoscopic conditions. This
study suggests a role for aOCT in human lower airway calibre measurements.
Supported by NHMRC scholarship 463926.
No conflict of interest.
TP 135
LUNG CANCER MANAGEMENT IN SOUTHERN TASMANIA

T CAMPBELL1, R HARLE1, N HARKNESS2
Department of Diagnostic Imaging, Royal Hobart Hospital, TAS 7000, and
2
Department of Respiratory Medicine, Royal Hobart Hospital, TAS 7000
TP 136
Background Lung cancer incidence and mortality are high in Tasmania.
MARKERS OF OXIDATIVE STRESS IN EXHALED BREATH OF

SUBJECTS WITH LUNG CANCER
HP CHAN1,2, V TRAN1,2, C LEWIS1,3, P THOMAS1,2
Faculty of Medicine, University of New South Wales, and Departments of
2
Respiratory Medicine and 3Medical Oncology, Prince of Wales Hospital,
Randwick, NSW 2031
Australia (AIHW 2003)

Tasmania (Cancer Registry 2003)
Incidence
Mortality
85/100 000
102/100 000
72/100 000
89/100 000
Aims and Objectives (a) to determine patient demographics in Southern

Tasmania, (b) to determine compliance to identified measures of best practice
and (c) assess referral rates, clinical utility and potential delay to Positron
Emission Tomography (PET) in a regional setting.
Methods A prospective database collected information on local clinical practice. Cases presented at a multidisciplinary lung cancer meeting over a 12
month period (March 2006April 2007) were analysed. Data were available for
n = 121/161 (75%). Results are shown as mean SD.
Results 113 primary lung cancer cases were identified. The mean age was
71 11 years. 58% of patients were male and 95% were current or
ex-smokers. 81% were non-small cell lung cancers (NSCLC).
% compliance to guidelines
Tissue diagnosis
Time from diagnosis to surgery (27 15 days)
Macroscopically complete surgical resection (9/11)
PET for Stage IIIb before radical chemoradiotherapy
93%
82% < 42 days
82%
75%
62% of patients presenting with early or locally advanced disease underwent

further staging with PET (n = 34/55). Management was changed in 50% of
cases (17/34). The average time from PET referral to scan was 11 5 days.
Conclusion A disproportionate number of lung cancers occurred in women.
Although surgery was performed within recognized timeframes, 2 of 11 patients
had incomplete resections. PET influenced management decisions and was
performed in a timely fashion.
Exhaled breath condensate (EBC) is a simple, safe and non-invasive method

of sampling breath and has the potential to investigate lung cancer and the
associated neoplastic process in the lungs. Increased oxidative stress has
been implicated in the pathogenesis of lung cancer, and is characterized by
elevated hydrogen ions, and hydrogen peroxide (H2O2), which is formed from
the conversion of superoxide anions by superoxide dismutase. Airway pH has
already been shown to be decreased in EBC of patients with other respiratory
conditions, but not in lung cancer. Therefore the concentration of H2O2 and
hydrogen ions in the EBC of lung cancer subjects was compared with matched
controls.
Methods Six subjects with newly diagnosed lung cancer were recruited and
matched with control subjects: non-smokers, ex-smokers and smokers. EBC
was collected and H2O2 was then measured by an assay method based on
oxidation of 3,3,5,5-tetramethybenzidine by horseradish peroxidase and H2O2
while pH was measured using a pH meter.
Results There was a significant difference (p = 0.033, ANOVA) in H2O2 concentration between the 4 groups with the lung cancer group having elevated
mean H2O2 concentration of 23.68 mM (9.15 (SEM) compared to the controls:
non-smokers, 17.59 mM (6.53 (SEM); ex-smokers, 14.35 mM (3.79 (SEM); and
smokers, 5.21mM (0.69 (SEM). pH did not differ significantly (p = 0.659,
Kruskal-Wallis test) between the groups.
Conclusion These preliminary data suggest that there is significant difference in H2O2 concentration between the groups. The demonstration of an
elevated H2O2 level in those with lung cancer indicates an increase in oxidative
stress which implies that this may be part of the pathogenesis or response to
neoplasia.
Supported by None.
TP 137
IMMUNE ESCAPE OF HUMAN LUNG CANCER CELLS BY

INHIBITING TH1 RESPONSE AND INDUCING INFLAMMATORY
CYTOKINES
G HODGE1,2, G MATTHEWS1, M HOLMES1, P REYNOLDS1, S HODGE1
Department of Thoracic Medicine, Royal Adelaide Hospital, Adelaide, and
2
Haematology Department, Womens and Childrens Hospital, North
Adelaide
Pro-inflammatory Th1 cytokines produced by T cells and monocytes play an

important role in the immune response to malignant cells. However, tumours
may escape immune surveillance by inhibiting Th1 response and promoting
chronic inflammation at the tumour site.
Methods To investigate the effect of soluble factors released by lung cancer
cells on T cell and monocyte pro- and anti-inflammatory cytokines, culture
supernatants from several lung cancer cell lines and a normal epithelial cell line
(16HBE) were cultured with whole blood for 24 hours, then for a further 16 hrs
with and without stimuli. Intracellular cytokine / chemokine production was
determined using multiparameter flow cytometry.
Results In stimulated cultures, there was a significant decrease in T cell Th1
pro-inflammatory cytokines IFNg, TNFa and IL-2 and a decrease in monocyte
IL-1a, IL-8, IL-12, TNFa, MCP-1 and MCP-3 but an increase in antiinflammatory cytokine IL-10 compared with 16HBE and control media. In
non-stimulated blood cultures there was an increase in all monocyte inflammatory cytokines / chemokines in the presence of lung cancer supernatants.
Conclusions Lung cancers secrete soluble factors that inhibit the antitumour pro-inflammatory Th1 response by T cells and monocytes and upregulate monocyte anti-inflammatory cytokine IL-10 following antigenic challenge.
Lung cancer cells may also escape immune surveillance by secreting soluble
factors that cause newly recruited monocytes to release inflammatory cytokines promoting chronic inflammation at the tumour site.
A59
TP 138
IMMUNE EVASION BY LUNG CANCER CELLS: POTENTIAL

ROLE OF PROTEINASE INHIBITOR-9
GM MATTHEWS, J AHERN, G HODGE, M HOLMES, H JERSMANN,
PN REYNOLDS, S HODGE
Department of Thoracic Medicine, Royal Adelaide Hospital, and Lung
Cytotoxic T-cells (CTLs) are important barriers against tumour cells. CTLs
induce apoptosis of target cells by mechanisms that include the release of
pore-forming perforin and granule associated enzymes, such as granzyme B
and granulysin. Proteinase inhibitor-9 (PI-9) is the only known granzyme B
inhibitor and its expression has been observed in some cancers. We hypothesized that PI-9 would be differentially expressed in lung cancer cells and may
inhibit granzyme B-induced apoptosis in these cells.
Methods We investigated PI-9, granulysin and granzyme B expression in
various lung cancer cell lines (1299, 1466, 2009, SBC-1) and normal epithelial
cells obtained from bronchial brushing using flow cytometry. Peripheral bloodderived T-cells were then incubated with lung cancer cell line supernatants and
levels of PI-9, granzyme B and T-cell reactive oxygen species (ROS) were
assessed.
Results PI-9 expression was detected in all lung cancer cell lines, (1299
(54.2%), 1466 (90.2%), 2009 (85%), SBC-1 (81%)), at much higher levels than
in normal bronchial epithelial cells (8.5%). Granzyme B and granulysin levels
were undetectable or low in cancer cells (09.2%). Increased expression of
PI-9 and reduced levels of granzyme B were observed in CD8+ T-cells in the
presence of all cancer cell supernatants tested (p < 0.05). Interestingly, T-cell
ROS levels were significantly increased in CD8+ T-cells after incubation with
cancer cell supernatants (p < 0.05).
Conclusions High PI-9 expression in lung cancer cells combined with a
reduction in T-cell granzyme B expression and enhanced intracellular T-cell
ROS levels may be a mechanism of immune evasion of lung cancer cells to
granzyme B-induced cytotoxicity.
TP 139
SYNERGISM BETWEEN IMMUNOTHERAPY AND

CONVENTIONAL CHEMOTHERAPY AND SURGERY FOR LUNG
MALIGNANCIES
BWS ROBINSON, AJ CURRIE, M BROWN, R LAKE, A NOWAK
School of Medicine and Pharmacology, University of Western Australia,
Nedlands, WA 6009 & National Research Centre for Asbestos Related
Diseases, Nedlands, WA 6009
Immunotherapy for lung malignancies such as lung cancer and mesothelioma
is most likely to be successful it it can be combined with conventional tumour
debulking approaches such as chemotherapy and surgery. But they scientific
basis of such combinations is yet to be determined.
To study this we evaluated (1) the capacity of different lung chemotherapy
drugs to alter tumour antigen cross-presentation and immunogencity, (2) duration of antigen presentation and responsiveness to immunotherapy after
debulking surgery with/without lymphadenectomy, and (3) the pattern of TLR
agonism which best synergized with chemotherapy and surgery.
We used the AB1-HA murine model of lung malignancy in BALB/c mice.
Results (1) The antimetabolite drugs gemcitabine and pemetrexed were
most immunogenic compared to the cytotoxic antibiotics doxorubicin and mitomycin C and the alkylating agent cisplatin. Gemcitabine delived large amounts
of tumour antigen into the cross-presentation pathway. (2) Tumour antigen
cross-presentation persisted for only 10 days following resection. The optimal
window for immunotherapy following cancer surgery is 1 week for effector CTL
stimulation and 24 weeks for memory CTL stimulation. (3) The viral-like TLR
agonists TLR 3, 7 and 9 were the most effective adjuvant TLR molecules, with
TLR 7 agonists generating the strongest systemic anti-tumour responses.
Conclusion These results help explain previous lung immunotherapy failures and will inform new clinical trials.
TP 140
SCREENING FOR MESOTHELIOMA USING SERUM V URINE

MESOTHELIN LEVELS
J CREANEY, N BOUDVILLE, AW MUSK, A SEGAL, BWS ROBINSON
School of Medicine and Pharmacology and Australia and National Research
Centre for Asbestos Related Disease, University of Western Australia,
Verdun St, Nedlands, W Australia 6009
Background Mesothelioma is a highly aggressive tumour with an increasing
world wide incidence. The serum biomarker mesothelin is elevated in some
individuals prior to development of clinical symptoms of the disease and may
be useful for screening. We therefore studied the sensitivity and specificity of
urinary versus serum levels of mesothelin for mesothelioma patients and
evaluated the influence if renal function on the biomarker level.
Materials and Methods Concurrent sera and urine samples collected from
patients with and control populations. Mesothelin concentrations were determined by double-determinant ELISA using the MESOMARKTM assay (FDI, PA).
Their estimated glomerular filtration rate (eGFR) was also calculated.
Results Mesothelin levels correlated between serum and urine samples
(Pearsons correlation 0.791; p < 0.0001). Mesothelin levels were significantly
higher in patients with mesothelioma compared to those with asbestosis and/or
pleural plaques in serum (4 0.9 versus 0.9 0.05 nM; p < 0.0001, respectively), in urine (1.9 0.5 versus 0.3 0.03; p < 0.0001) and in urine following
normalization using creatine levels (0.2 0.05 versus 0.04 0.01). Age and
eGFR were significantly associated with mesothelin levels.
Conclusion The sensitivity and specificity of mesothelin in urine and in
serum were comparable. Urine mesothelin may prove to be a useful alternative
to serum mesothelin for mass screening of asbestos-exposed individuals.
A60
TP 141
INCIDENTAL LUNG FINDINGS ON CT CORONARY ANGIOGRAM

AND THEIR CLINICAL SIGNIFICANCE
T Sather1, M Hansen2, A Khoo2, Fong K1, Slaughter R2
Dept. of Thoracic Medicine, The Prince Charles Hospital, Chermside QLD
4034, and 2Dept. of Radiology, Prince Charles Hospital, Chermside QLD
4034
TP 142
ANALYSIS OF BIOMARKERS IN EXHALED BREATH OF

PATIENTS WITH LUNG CANCER
VH TRAN1,2, C LEWIS3, P JACKSON4, PS THOMAS1,2
School of Pathology, School of Medical Sciences, UNSW, and Departments
of 2Respiratory Medicine, 3Medical Oncology, and 4Oncology Research
Centre, Prince of Wales Hospital, Randwick, NSW 2031
Patients undergoing CT Coronary Angiogram (CTA) are often former or current

smokers with a high incidence of asymptomatic lung disease. Overseas reports
show a rate of lung abnormalities ranging from 6.7% to 19%. There are no
studies from Australia and local factors such as the higher incidence of atypical
Mycobacteria may influence the rate of benign findings. We are therefore
performing a prospective observational study to identify the prevalence and
characteristics of incidental lung findings in people undergoing routine CTA.
Methods Population: 100 patients undergoing routine CTA after informed
consent. Intervention: Radiologist evaluation of lung windows on diagnostic
standard workstations. Comparator: Uncontrolled observational study of consecutive patients. Outcomes: Primary: Prevalence and characteristics of
abnormal findings, final diagnosis (clinical judgment, biopsy or long term followup). Secondary: Number of downstream investigations and costs.
Results 25 CTAs have been studied to date. In 8/25 (32%), abnormalities
were noted on lung windows. In 2/25 (8%), there were lung nodules, in 2/25
(8%) there were hilar lymph node abnormalities, in 1/25 (4%), there was
hemidiaphragm elevation and in 3/25 (12%) there were pleural plaques (data
collection ongoing with study closure expected in February 2008).
Conclusions Preliminary data indicate a substantial number of incidental
pulmonary findings from CTA; full results will be presented. Further analysis is
required to determine the impact (benefits, costs and harms) that may result
from the concurrent examination of lung windows at routine CTA.
Aim Increased levels of nitrogen oxides (NOx) and inflammatory markers

have been found in bronchoalveolar fluid of lung cancer (LC) patients, but have
not been investigated in exhaled breath condensate (EBC).The aim of this
study was to compare NOx and total protein levels in EBC of LC patients with
control subjects.
Methods EBC was collected during tidal breathing through a glass collection
device cooled to 4C. EBC NOx concentrations were measured by a fluorescent modification of the Greiss method. Total protein in EBC was determined
employing the bicinchoninic acid (BCA) assay. EBC NOx data were log transformed. All data were analysed using ANOVA and expressed as mean SEM.
Results A total of 88 control subjects and 54 patients with primary LC were
recruited. NOx and protein concentrations are shown in Table 1.
EBC NOx
EBC Total Protein
Subject Group
NOx
Total
Non-Smoker
Ex-Smoker
Smoker
Respiratory Conditions
Lung Cancer
25
19
13
31
54
19.6 2.2 mmol/L

29.8 6.1 mmol/L
23.5 1.9 mmol/L
23.9 2.1 mmol/L
24.6 2.8 mmol/L
13
6
6
16
28
11.3 0.6 mg/mL

10.9 0.4 mg/mL
12.5 0.7 mg/mL
11.0 0.4 mg/mL
13.8 0.8 mg/mL
There was no significant difference in EBC NOx levels (p > 0.05), but in total
protein there was a significant difference between lung cancer patients and all
control groups (p = 0.04).
Conclusion Significantly increased EBC total protein levels were found in
patients with lung cancer. These data suggest that protein mediator secretion
or vascular leak may be present in those with lung cancer. Future studies will
focus upon the identification of these proteins.
TP 143
GENE BASED RISK SCORE IDENTIFIES SMOKERS AND

EX-SMOKERS AT HIGH RISK OF LUNG CANCER
OLIV SIG Poster Session
R YOUNG1,4, R HOPKINS1, B HAY1, M EPTON3, P BLACK1, H GARDNER1,

R SULLIVAN2, G GAMBLE1
Departments of 1Medicine and 2Oncology, Auckland Hospital, Auckland,
3
Department of Medicine, University of Otago, PO Box 4345, Christchurch,
and 4Synergenz Biosciences Ltd, PO Box 37-971, Auckland, NZ
LEFLUNOMIDE-INDUCED INTERSTITIAL LUNG DISEASE (ILD)

AND GLOMERULONEPHRITIS (GN)
Exposure to cigarette smoking is central to the development of lung cancer.

Other risk factors include increasing age, impaired lung function, family history
of lung cancer and asbestos exposure.
Methods In this two stage case-control study 446 lung cancer cases and 484
healthy smoker controls were recruited. 180 genetic markers (SNPs) implicated in lung cancer were screened in our test cohort of 439 smokers and
ex-smokers. 30 SNPs whose genotypes (co-dominant or recessive model)
were associated with either the healthy smokers (protective) or lung cancer
(susceptibility) phenotype were identified. After genotyping this 30 SNP panel
in a second cohort of 491 subjects 19 SNPs were chosen and assigned a
simple composite genetic score that was combined with scores for age, history
of COPD and family history of lung cancer, weighted according to our multivariate regression analysis (n = 930 total subjects).
Results The lung cancer risk score was linearly related to the likelihood of
lung cancer with odds ratios (referenced against the lowest score quintile)
ranging from 1 to 29 in the highest quintile. On receiver operator curve analyses, the AUC was 0.78 and the frequency distribution showed bimodal separation between healthy smokers and lung cancer cases. Utility of the score was
not affected by effects of age, smoking history or lung function.
Conclusion We suggest that genetic data may be combined with other risk
variables to define smokers or ex-smokers at risk of lung cancer for targeted
interventions such as smoking cessation and early detection of lung cancer.
Supported by Health Research Council, NZ.
TP 144
V AIYAPPAN1, A GRAHAM2
Department of Medicine, Maroondah Hospital, Melbourne, Australia, and
2
Department of Medicine, Huddersfield Royal Infirmary, Huddersfield, UK
The new disease-modifying anti-rheumatic drug (DMARD) leflunomide is being

used increasingly to treat inflammatory arthritis. Its association with interstitial
lung disease needs to be considered before combining it with methotrexate.
Case Report A 73-year-old male who was known to have Rheumatoid arthritis and was on methotrexate was admitted with progressive dyspnoea and
malaise. He had been recently started on leflunomide. Investigations revealed
interstitial lung disease and acute renal failure. He improved on conservative
treatment (stoppage of disease modifying drugs (DMARD), IV fluids and steroids).
Review of Literature An epidemiological study by Suissa et al has suggested that there is increased risk of ILD associated with leflunomide in
patients with a history of ILD or methotrexate use but they attributed this to
channelling bias. There has also been a report of leflunomide associated with
IgA glomerulonephritis.By this presentation we aim to increase the awareness
of this entity. We also suggest that any patient who is started on combination
DMARD (i.e. methotrexate and leflunomide) should have a baseline Chest
X-ray and be monitored for development of interstitial lung disease.
Conclusion We are reporting the first ever case of Interstitial lung disease
and Glomerulonephritis (in the same patient), due to usage of leflunomide. This
entity needs to be thought about in any patient on combination DMARDs.
TP 145
TP 146
BONE MORPHOGENETIC PROTEIN (BMP) RECEPTOR-II AND

BMPS ARE DOWN-REGULATED IN PATIENTS WITH
PULMONARY EMBOLISM
INTRAVENOUS CYCLOPHOSPHAMIDE IN PROGRESSIVE

FIBROTIC LUNG DISEASE: REVIEW OF A TREATMENT
STRATEGY
L BECKERT1, S GUNNINGHAM2, P CHIN1, B ROBINSON1

Depts. of 1Med and 2Path, University of Otago Christchurch, Chch 8001, NZ
Background Bone Morphogenic Protein Receptor II (BMPR-II) mutations are

associated with pulmonary artery hypertension. Failure of the growth inhibitory
effects of BMP may contribute to vascular obliteration and remodelling leading
to pulmonary artery hypertension (PAH) [1]. PAH has been observed following
venous thrombembolic disease (VTE), including pulmonary embolism (PE) and
deep venous thrombosis (DVT) [2]. Local markers of the pulmonary vascular
endothelium rather than traditional markers of thromobophilia are thought to be
involved [3].
Methods Plasma was collected from age and gender matched participants
within 24 hours of diagnosis of VTE and prior to commencement of Warfarin
therapy. Plasma samples were hybridized to individual human cytokine antibody arrays, to detect protein levels of BMP2, BMP4 and BMPR-II.
Results BMP2 and BMP4 levels were higher in patients with DVT than PE.
No difference in the BMP level was observed between patients with PE and
controls. Soluble BMPR-II receptor was lower in patients with PE than in
controls or patients with DVT.
Conclusion In patients with pulmonary artery stress during the time of a PE
the BMPR-II receptor is reduced, which may predispose patients to vascular
remodelling and obliteration. The BMP 2 and 4 levels are reduced at the same
time, suggesting a possible overriding regulatory mechanism. The physiological role of BMPs and BMP receptors in patients with VTE warrants further
investigation.
References
1. Jeffery, TK, et al. Pro Cardiovascular Disease 2002;45:173202.
2. Pengo V, et al. N Engl J Med 2004;350:22572264.
3. Lang I et al. Proc Am Thorac Soc 2006;3:568570.
Supported by Canterbury District Health Board, University of Otago and Lottery
Health.
Nomination None.
A61
TJ CORTE1, E RENZONI1,2, R DU BOIS1,2, R ELLIS1, AU WELLS1,2

Royal Brompton Hospital, 2Imperial College, London, United Kingdom
Historically, cyclophosphamide has had a variable role in interstitial lung

disease (ILD), the rationale for its use based on the benefit seen in vasculitis
and scleroderma, its rapid effect and low toxicity profile. In patients with severe
progressive ILD a rapidly effective, well-tolerated agent is desirable. For this
reason a treatment protocol for the use of intravenous (IV) cyclophosphamide
was implemented at our hospital.
Aim To review the indications, duration, tolerability and effect of intravenous
cyclophosphamide in ILD patients following the introduction of a treatment
protocol.
Methods Records of 92 patients [DLco was 40 15% and FVC 61 20%]
completing a course of IV cyclophosphamide during 20056 were reviewed
(excluding patients with systemic sclerosis). Data covering 18 months prior to
and following treatment were collected. Comparative analysis of paired pulmonary function data 6 months before and after treatment was performed. 61%
had underlying autoimmune disease.
Results Primary treatment indications included progressive disease(n = 67);
severe disease (n = 16); suspected vasculopathy (n = 11); bridging therapy to
transplantation (n = 10); and accelerated decline (n = 5). Patients received
600 mg/m2 [mean dose 1152 165 mg, median number of pulses 6 (112)].
Patients with paired pulmonary function data had a difference in median
change in DLco% predicted from -15.6% (-95.4 to 29.9%) before treatment to
+4.25% (-17.3 to 73.9%) following treatment (p < 0.0001). This remained significant with exclusion of vasculitis, or any autoimmune disease, and independent of prior immunosuppression. Therapy was well tolerated (4 withdrew from
treatment, 5 deaths within 1yr, none directly related to treatment).
Conclusion IV cyclophosphamide is well tolerated, and associated with functional stability or improvement in the majority of patients. It remains a viable
treatment alternative for consideration.
TP 148
TP 147
PULMONARY HYPERTENSION(PH) IN FIBROSING LUNG

DISEASE: NON-INVASIVE MEASURES COMPARE TO RIGHT
HEART CATHETERIZATION
TJ CORTE, SJ WORT, M GATZOULIS, AU WELLS
Royal Brompton Hospital, London, UK
Pulmonary hypertension is common in interstitial lung disease (ILD) and associated with a poor prognosis. As the gold-standard test, right-heart catheterization (RHC) is invasive, and resource-limited, reliable non-invasive measures
of PH are needed.
Methods All ILD patients referred for RHC during 19972007 were included
(n = 95; 54 male; age 56.5 12 yrs). All patients had concurrent echocardiography (TTE) and pulmonary function. The relationship of RHC mean pulmonary artery pressure (mPAP) to TTE variables, pulmonary function, exercise
capacity, as measured by six minute walk testing (6MWT, n = 58) and brain
natriuretic peptide (BNP, n = 36), was examined.
Results 64 of 95 patients had PH on RHC [mean mPAP 32.2 12.7 mmHg,
pulmonary vascular resistance (PVR) 5.7 4.3 WU, cardiac output
4.3 1.2 L/min] and TTE [RVSP 57 19 mmHg (n = 70), pulmonary acceleration time (PAT) 97.6 27.6 ms (n = 65)]. Patients had functional impairment (DLco 31.6 18.2%, PaO2 8.5 2.5 kPa, desaturation 88% on
6MWT in 81%). BNP levels were 19 pmol/L (1.4342).
mPAP correlated with TTE parameters [RVSP (r = 0.53, p < 0.0001), peak
tricuspid velocity (r = 0.46, p < 0.01), degree of tricuspid regurgitation (r = 0.33,
p < 0.01), PAT (r = -0.35, p < 0.01), resting pulmonary function [DLco/VA
(r = -0.22, p = 0.03), PaO2 (r = -0.35, p < 0.001)] and BNP (r = 0.40, p = 0.02).
No correlation was found with 6MWT variables. On stepwise linear regression,
RVSP (p < 0.0005) and PaO2 (p < 0.01) were independent determinants of
mPAP (equation r2 = 0.33).
Conclusion PH has important prognostic and potential therapeutic implications in patients with fibrotic lung disease. mPAP on RHC, correlates with TTE
measures of PH, as well as DLco/VA, PaO2 and BNP. These results have the
potential to guide physicians in the selection of patients referred for RHC.
MASS SPECTROMETRY: A NON-INVASIVE MEASURE OF

PULMONARY BLOOD FLOW IN PATIENTS WITH PULMONARY
HYPERTENSION (PH)
TJ CORTE, C HARRIES, D CRAMER, S WARD, M GATZOULIS,
AU WELLS
Respiratory Mass Spectrometry (RMS) is a simple, non-invasive and reproducible measure of pulmonary haemodynamics, including pulmonary blood
flow (QPF). In patients with PH where repeated right-heart catheterization
(RHC) is not routinely performed, a non-invasive measure of QPF is a desirable
tool for clinical practice.
Methods All patients with PH referred for RMS during 20067 were included
(n = 15). All patients also had concurrent pulmonary function, echocardiography, and submaximal exercise test data, and 14 had concurrent BNP levels.
Relationships between QPF and other non-invasive variables indicating pulmonary vascular dysfunction were examined.
Results All patients had PH on echocardiogram [mean tricuspid gradient
(TG, n = 8) was 90 27 mmHg, pulmonary acceleration time (PAT, n = 8) was
56.6 16.6 ms]. Underlying aetiology for PH included Eisenmenger syndrome
(n = 11), idiopathic pulmonary arterial hypertension (n = 2), chronic thromboembolic PH (n = 1), and HIV-related PH (n = 1). Mean RMS QPF was
2.9 0.7 L/min and DLco was 65.9 13.5%, FVC 79.9 20.8%, and median
BNP 23.5 (7.5 to 88.4). Exercise capacity was impaired: median World Health
Organization (WHO) functional class 2; and mean six minute walk testing
(6MWT) minimum oxygen saturation was 70 13%.
QPF correlated with DLco (r = 0.56, p = 0.03), resting pulse oximetry (r = 0.67,
p = 0.01), and 6MW minimum saturation (r = 0.78, p < 0.01). No significant
correlation was found between QPF and echocardiographic parameters, BNP
or 6MW distance.
Conclusion RMS measurement of QPF is a simple, reproducible and noninvasive, and correlates with other non-invasive indices of PH suggesting its
usefulness in clinical practice. Further testing is required to determine its
relationship with echocardiographic and right heart catheter measures of PH
severity.
A62
TP 149
BRAIN NATRIURETIC PEPTIDE (BNP) CORRELATES TO

ECHOCARDIOGRAPHIC INDICES OF RIGHT VENTRICULAR
FUNCTION IN PATIENTS WITH INTERSTITIAL LUNG DISEASE
(ILD)
TJ CORTE, SJ WORT, R ENGEL, G GIANNAKOULAS, AU WELLS
Pulmonary Hypertension (PH) contributes to increased mortality and morbidity
in ILD. In ILD, elevated BNP is associated with PH on right heart catheterization
(RHC), however the relationship with echocardiographic (TTE) PH indices is
not established.
Methods All patients with ILD with BNP concentrations performed during
20057 were included (n = 91, 51 male, age 64 16 yrs). All had concurrent
pulmonary function, 54 had 6minute walk tests (6MWT) and 16 had RHC. TTE
tapes were reviewed by an independent cardiologist blinded to patients other
results. Relationships between BNP concentrations and TTE measures of PH
were studied.
Results 42 had PH on TTE [tricuspid peak gradient (TRPG, n = 70)
40 19 mmHg, pulmonary acceleration time (PAT, n = 89) 98 27 ms]. BNP
was 10 pmol/L (1.4377). Patients had functional impairment [DLco
37.8 16.1%, 6MWT end SpO2 83 9%].
BNP correlated with TTE markers of right ventricular function: TRPG (r = 0.39,
p = 0.0008), PAT(r = -0.33, p = 0.001), right atrial pressure (r = 0.30,
p = 0.007), right atrial area (0.41, p = 0.0001), right ventricular inlet diameter
(r = 0.38, p = 0.003) and longitudinal anulus motion at the tricuspid valve
(r = -0.29, p = 0.008), but did not correlate with markers of left heart function.
BNP also correlated with DLco(r = -0.38, p = 0.0002), DLco/VA (r = -0.33,
p = 0.001), WHO functional class(r = 0.34, p = 0.001) and 6MW
distance(r = -0.34, p = 0.01). In the subgroup of patients with RHC, BNP correlated with mPAP (r = 0.56, p = 0.03).
Conclusion In patients with ILD, BNP concentrations correlate with echocardiographic indices of right heart dysfunction as well as other indirect measures
of vascular compromise. These non-invasive tools may be useful alone or in
combination in the diagnosis and monitoring of PH in ILD patients.
TP 150
ROLE OF ANTIOXIDANTS IN THE MANAGEMENT OF EARLY

IDIOPATHIC PULMONARY FIBROSIS: A PROMISING CASE
M DAULATZAI, K TONG
Camberwell Medical Clinic, Camberwell, Vic 3124
Case A 65 year old male, non-smoker for 25 years, retired professor of
anatomy (had chronic exposure to embalming fluids, formaldehyde, phenol,
antifungal and other solvents, for 20 years) presented with chronic cough and
phlegm production. These symptoms were worse at night (waking him several
times) and early morning. His pulmonary tests were stopped due to persistent
cough. A chest X-ray revealed features of longstanding interstitial lung disease.
The HRCT revealed widespread subpleural interlobular thickening, worse at
bases, in keeping with idiopathic pulmonary fibrosis (IPF). There was minimal
fibrosis and honeycombing, but no groundglass opacification, large bullae,
pleural calcification or pleural plaques. However, there was associated bronchiectasis at the lung bases considered to be due to traction. The BA lavage
showed 50% macrophages, 7% neutrophils, 3% Lymphocytes, and 40%, eosinophils and no infection. The patient declined to have a lung biopsy. As per his
past X-rays, the duration of his IPF is a little over one year. He maintains that
his symptoms started only after starting irbesartan (IRB).
Management and Comments The patient stopped taking IRB 6 months ago,
and concomitantly started taking a regimen of antioxidants (AO)Quercetin,
Curcumin, and Vitamins C and E. His BP is stable (140/88) on thiazide. For the
past 3 months, his cough and phlegm production are reduced by about 90%.
He now sleeps (8 hrs) undisturbed by cough. IPF is a chronic, progressive, and
often fatal form of interstitial lung disease. In IPF, the antioxidant/prooxidant
balance leads to increased oxidative stress and dysregulation of vital body
functions. AO quench free radicals, help regenerate other antioxidants, and
favourably affect anabolism and gene expression. This case underscores the
possibility of attenuation of this relentless fatal disease.
TP 151
TP 152
DOES HRCT PREDICT THE LIKELIHOOD OF A POSITIVE

TRANSBRONCHIAL BIOPSY IN SARCOIDOSIS?
PULMONARY HYPERTENSION COMPLICATING SARCOIDOSIS
S DE BOER1, DJ MILNE2, M OCARROLL1, ML WILSHER1

Green Lane Respiratory Services, and 2Radiology Department, Auckland
City Hospital, Auckland, New Zealand
B RHODES, M MUSK, D CHAMBERS, C JARY, E GABBAY

Advanced Lung Disease Unit, Royal Perth Hospital, Western Australia, 6008
Introduction Transbronchial lung biopsy (TBB) has a variable and unpredictable diagnostic yield in sarcoidosis. We hypothesized that the extent and
pattern of parenchymal disease on CT would predict the likelihood of a positive
TBB.
Methods Data relating to ethnicity, symptoms, pulmonary function and site
and results of TBB and bronchoalveolar lavage (BAL) from 70 sarcoidosis
patients were recorded. All had a CT scan within 6 weeks prior to the TBB
procedure. CXR stage was determined from radiology report. CT scans were
scored quantitatively for patterns of parenchymal disease (nodular, reticular,
consolidation, ground glass and mosaic attenuation) on a lobar basis.
Results 50% patients had a positive TBB (total 67% of cohort had histological confirmation). Symptoms, ethnicity, treatment, lung function and CXR stage
were not predictors of a positive biopsy. Positive biopsy was associated with
higher BAL lymphocyte count (p < 0.05) and female gender (p < 0.01). A reticular pattern (p < 0.05) and higher total lung score (excluding DA) (p < 0.05) on
CT scan predicted a positive biopsy. In those patients with TBB from right lower
lobe (53/70) the total RLL score on CT was predictive of positive biopsy
(p < 0.05). On multivariate analysis gender, BAL lymphocytosis and total lung
score were independent predictors of a positive TBB (area under ROC 0.82).
Conclusion The total extent of parenchymal disease on CT scan in addition
to the pattern and distribution predicts the likelihood of a positive TBB at
bronchoscopy.
Acknowledgements: Irene Zeng and Wendy Fergusson for statistical advice.
Supported by a grant from the Myrtle Martin Trust.
Pulmonary hypertension (PAH) is well documented in patients with advanced

sarcoid parenchymal disease, however it is increasingly recognized to occur in
those with minor parenchymal disease. A recent Australian review of sarcoidosis patients did not identify any patients with PAH.
Method We reviewed our population of patients at a PAH quaternary referral
centre and identified 6 patients who also had the diagnosis of sarcoidosis. The
pathway to diagnosis and effect on subsequent medical management was
reviewed.
Results Sarcoidosis was diagnosed before pulmonary hypertension in 3
patients. All had minor parenchymal disease with persistent dyspnoea leading
to the diagnosis of PAH. The degree of PAH was out of keeping with the degree
of parenchymal disease in all cases.
2 patients were diagnosed with pulmonary hypertension prior to the identification of sarcoidosis. One patient with exercise induced PHT underwent a lung
biopsy which demonstrated granulomatous infiltration and sarcoid angiitis.
A further patient underwent lung transplant for an end stage fibrotic lung
disease. PAH was identified at transplant work up and explant histology demonstrated sarcoidosis and pulmonary capillary haemangiomatosis.
3 out of 6 patients have been treated with specific PAH therapy resulting in
improvement in exercise tolerance and haemodynamics. 2 patients have yet to
commence PAH therapy and 1 patient underwent lung transplantation.
Conclusion This small cohort of patients represents the wide spectrum of
presentation of these dual conditions. Histopathology on 2 patients adds to the
emerging knowledge of the pathogenesis of PAH in sarcoidosis. Identification
of both conditions is important and relevant to tailoring medical therapy.
TP 153
A63
TP 154
FAILURE OF BOSENTAN IN PULMONARY ARTERIAL

HYPERTENSION
PRIMARY CUTANEOUS T CELL LYMPHOMA PRESENTING AS

PURITIC RASH, COUGH AND PROGRESSIVE DYSPNOEA
S HARCH, H WHITFORD, C MCLEAN

Department of Medicine, The Alfred Hospital, Victoria 3181
BENJAMIN CH KWAN1, SUSAN MACCALLUM2, GLENDA WOOD3,

DAVID MCKENZIE1
1
Department of Respiratory Medicine, Prince of Wales Hospital, Sydney,
NSW 2031, 2Department of Haematology, Prince of Wales Hospital, Sydney,
NSW 2031, and 3Department of Dermatology, Prince of Wales Hospital,
Sydney, NSW 2031
Pulmonary arterial hypertension has two histological variants; arterial-only

pulmonary arterial hypertension (artPAH) and pulmonary veno-occlusive
disease (PVOD). Bosentan, a dual endothelin receptor antagonist, has been
found to improve haemodynamics, functional capacity and survival in artPAH.
However, the response to Bosentan in clinically diagnosed artPAH is often
variable. It was hypothesized that a lack of response to Bosentan therapy in
clinically diagnosed artPAH can be explained by misdiagnosed PVOD. Aims
included to: (1) perform morphometric and qualitative pulmonary vessel analysis on normal controls and cases clinically diagnosed with artPAH who had
failed Bosentan therapy; (2) ascertain if PVOD is present within the case group;
(3) correlate clinical variables and vessel microanatomy to identify the pathologies driving pulmonary pressure elevation. This study reviewed 14 cases of
clinically diagnosed artPAH (idiopathic n = 12, associated with scleroderma
n = 2), who had failed Bosentan therapy and had available lung tissue. Controls
(n = 6) were obtained from explanted lungs for other causes and a prior
transthoracic echocardiogram excluded pulmonary hypertension. Vessel morphometry and qualitative analysis was performed with a novel technique of
smooth muscle actin immunohistochemistry counterstained with Verhoeffs
elastin. Baseline clinical data were retrieved. We found 86% of cases had
pathology confirmed PVOD. Only 14% of cases had artPAH, the original clinical
diagnosis. In PVOD, significant pathology was present in all vessel types. All
vessels had significant smooth muscle hypertrophy. The obstructive, collagenous, pauci-cellular intimal fibrosis of the venules (p < 0.0001) and arterioles
(p < 0.0001) was considerably different to the concentric laminar proliferation of
smooth muscle observed in the muscular arteries (p < 0.0001) and arterioles
(p = 0.001) in artPAH. artPAH also had muscular artery smooth muscle hypertrophy (p = 0.007). The median time to Bosentan failure was shorter in PVOD
than artPAH (290 vs. 657 days). In conclusion, PVOD is an under-diagnosed
cause of pulmonary hypertension, is commonly clinically misdiagnosed as
artPAH and may present with a poor Bosentan therapy response. Finally,
PVOD is a vaso-occlusive, not a veno-occlusive disease, and is an independent type of pulmonary hypertension, not a subtype of pulmonary arterial
hypertension.
Cutaneous T cell Lymphomas (CTCL) are a heterogenous group of lymphoproliferative disorders. They show various clinical manifestations and diverse
morphological, histological and immunological characteristics of the malignant
cells. They are caused by clonally derived, skin invasive T cells. Peripheral T
cell Lymphomas (PTCL) are generally more aggressive and have one of the
lowest overall and failure-free survival rates. Because of the rarity of these
disorders, diagnosis and treatment remain challenging. This case report
describes a 69-year-old woman presenting with progressive dyspnoea and
cough, together with a distressing generalized pruritic rash. She was initially
treated as left ventricular failure with the rash ascribed to a drug reaction as
suggested by initial skin biopsies. The diagnosis was made on a third skin
biopsy and flow cytometry of lymphocytes obtained by broncho-alveolar lavage
6 months after presentation. Despite an initial response to chemotherapy she
succumbed to the disease 20 months after diagnosis.
TP 156
TP 155
PRE-TEST CLINICAL PROBABILITY OF PULMONARY EMBOLUS

(PE) AND THE APPROPRIATE USE OF CTPA IN A TEACHING
HOSPITAL
V MALIPATIL1, P COUGHLIN2, D LEACH3, F THIEN1
Departments of 1Respiratory Medicine, 2Haematology and 3Emergency
Medicine, Box Hill Hospital and Monash University, Box Hill, Victoria 3128
Background Clinical risk stratification of PE is important to reduce indiscriminate investigation and improve interpretation of results. Increasing availability
of computed tomography pulmonary angiography (CTPA) has meant this
modality is a frequent first line investigation. Assessment with established
models of pre-test clinical probability for PE is often inadequate. Consequently
many patients undergo unnecessary investigation with CTPA.
Methods Retrospective audit of medical records for clinical reasoning in
ordering CTPA and assessment of pre-test clinical probability of PE according
to Wells score. Clinical risk according to objective criteria was recorded. For the
subjective component entitled An alternative diagnosis is less likely than PE,
the medical notes of clinical assessment were independently reviewed by three
senior clinicians and scored according to majority consensus. D-Dimer level
was also recorded.
Results Series of 154 consecutive CTPAs resulted in 15 (10%) positive for
PE. First 20 studies were reviewed. Patients from emergency department
(13/20), inpatients and outpatients were included. Applying Wells score, the
majority of patients had low pre-test clinical probability of PE (14/20). Only 1
case had high pre-test probability, but negative CTPA. One (1/20) CTPA was
positive for PE with intermediate pre-test probability. Additional observation
was that, when measured, an elevated D-Dimer was often the clinicians main
documented indication for ordering CTPA.
Conclusion Using Wells score, most patients investigated with CTPA have a
low pre-test clinical probability for PE. Often an elevated D-Dimer level has led
to investigation with CTPA despite low clinical risk. Application of clinical predictive models and better understanding of the D-Dimer assay may lead to
improved utilization of CTPA. Retrospective analysis of the rest of the cohort is
underway.
LUNG SINGLE PHOTON EMISSION COMPUTED TOMOGRAPHY

(SPECT)A USEFUL TOOL FOR DIAGNOSING PULMONARY
EMBOLISM (PE)
S MILES1, K ROGERS1, P THOMAS1, B SOANS1, J ATTIA1,2, C ABEL1,
E HOLT1, C DESTE2, M HENSLEY1,2
1
John Hunter Hospital, New Lambton, NSW 2310, and 2University of
Newcastle, Callaghan, NSW 2308
Improvement in nuclear medicine imaging of the lung through SPECT offers
the possibility of a better alternative to computed tomography pulmonary
angiography (CTPA) than planar ventilation-perfusion (V/Q) scan for the diagnosis of PE.
Methods This prospective study assessed the value of SPECT for the diagnosis of PE, by comparing SPECT to CTPA (using a 16-slice CT scanner) in
100 patients referred for investigation of suspected PE. The patients had CTPA
and V/Q lung scans (planar and SPECT) within 24 hours of each other. The
study assessed the value of SPECT by: (i) its sensitivity and specificity compared with a gold standard final diagnosis made by a clinician panel provided
with all information, including CTPA; (ii) direct comparison of SPECT and CTPA
results using kappa scores; (iii) comparison, using Kappa, of SPECT and CTPA
to a panel-based consensus clinical diagnosis of PE derived using all data
except SPECT and CTPA; (iv) comparison of inter-rater variability in SPECT
and in CTPA, each reported by two blinded readers.
Results (i) VQ imaging was performed for 87 patients, producing technically
adequate SPECT results in all cases, with a sensitivity of 0.83 and a specificity
of 0.98 compared to the gold standard.( ii) Agreement between CTPA and
SPECT was very good at 94% (Kappa = 0.87). (iii) Agreement between
the consensus clinical diagnosis and each test was similar: SPECT
(Kappa = 0.76); CTPA (Kappa = 0.77). (iv) There was better inter-rater agreement for duplicate SPECT reports (Kappa = 0.83) than for duplicate CTPA
reports (Kappa = 0.71).
Conclusion In this study, there is good evidence that SPECT has a similar
performance to CTPA as a test for the diagnosis of PE. SPECT has the
advantage of a lower dose of radioactivity and contrast but it does not provide
the radiological imaging of the lungs and chest obtained with CTPA.
No external support
A64
TP 157
EMERGENCY DEPARTMENT BASED CLINICAL

PATHWAY FOR SUSPECTED PULMONARY
EMBOLISM: A PROSPECTIVE STUDY
TP 158
A CASE OF RECURRENT SPONTANEOUS PNEUMOTHORAX IN

HIGH RISK PREGNANCY
B NG1, S LINDSTROM1, D DWYER2, C MACLEAN2

Department of Respiratory and 2Emergency Medicine, St George Hospital,
Kogarah, NSW 2217
Clinical pathways to guide the investigation of suspected pulmonary embolism

(PE) have been increasingly adopted by emergency departments (ED) worldwide. Compliance with these diagnostic algorithms is critical in ensuring good
patient outcomes. This study evaluated the compliance to the clinical pathway
used in our ED that combines risk assessment (Wells scoring system) with
D-dimer test, VQ scan or CTPA. The main objectives of this study were to
identify those factors which contributed to compliance and to assess patient
outcomes.
Methods A prospective observational study of 239 consecutive patients who
underwent investigation for PE in our ED. Patient demographics, pathway
parameters and patient outcomes at 3-month follow-up were collected.
Results Incidence of PE was 8.4% (n = 20). PE was diagnosed in 3 other
subjects (1.38%) within the follow-up period. Compliance to the clinical
pathway occurred in 120 subjects (50.2%). Non-compliance occurred in 71
subjects (29.7%). 48 subjects (20.1%) underwent appropriate risk assessment
but subsequent diagnostic tests did not conform to the stated pathway (partial
compliance). Compliance in documenting an initial risk assessment was greatest in subjects assessed by junior ED doctors (OR 1.97, p < 0.05). Noncompliance with this process was more likely to occur in those subjects with
active malignancy (OR 2.93, p < 0.01). Non-compliance to the pathway led to
unnecessary tests being performed more often (OR 2.36, p < 0.01) and CTPAs
being requested more frequently (OR 2.36, p < 0.005).
Conclusion Junior ED doctors were more likely to use a clinical pathway for
suspected PE. However, non-compliance with this pathway was common and
was associated with an excess of unnecessary investigations.
R PEARSON, E STONE
Department of Thoracic Medicine, St Vincents Hospital, Darlinghurst, NSW
2010
Case We report the case of a 37 year old woman who presented to the
Emergency Department with a three day history of dry cough and dyspnoea.
The patient was in her third pregnancy at 30 weeks gestation. She had no
fever, chest pain or coryzal symptoms. The patient had presented with a right
sided spontaneous pneumothorax seven months prior to the current presentation. Her past medical history included placental abruption, complicating her
previous two pregnancies. Her second pregnancy was complicated by placental abruption at 27 weeks and the foetus had not survived. Her first pregnancy
was complicated by placental abruption at 36 weeks with successful delivery of
the foetus. At presentation, significant findings included tachycardia, hypoxemia, tachypnoea and reduced breath sounds over the right side of the chest.
Chest x-ray demonstrated a large right pneumothorax. A right intercostal catheter was inserted resulting in right lung re-expansion. The catheter was
removed three days later. The patient returned to hospital twenty four hours
after catheter removal with a recurrent right sided pneumothorax. The patient
agreed to surgical intervention involving video-assisted thoracotomy and talc
pleurodesis. The patient had no further complications with the pregnancy. She
delivered a healthy baby at 38 weeks gestation.
Discussion Spontaneous pneumothorax in pregnancy is rare and there is
little evidence to provide guidelines for the management of recurrent pneumothorax in high risk pregnancy. Our case illustrates a successful outcome for
mother and foetus with surgical intervention at 32 weeks gestation.
TP 160
VARIATION IN CLINICAL PRACTICE BY RESPIRATORY

PHYSICIANS AND RHEUMATOLOGISTS MANAGING
SCLERODERMA LUNG DISEASE
TP 159
INTERSTITIAL LUNG DISEASE FOLLOWING TREATMENT WITH

OXALIPLATIN, 5-FLUOROURACIL AND LEUCOVORIN (FOLFOX)
F PICCOLO1, A TRIBE1, P THOMPSON1,2
Department of Respiratory Medicine, and 2Lung Institute of Western
Australia, Sir Charles Gairdner Hospital, Perth, WA 6009
1
FOLFOX is currently the standard adjuvant treatment for locally advanced

(stage III) colon cancer and increases disease free survival. Its toxicity is well
tolerated with common adverse effects being paraesthesia, bone marrow suppression and gastrointestinal disturbance. Pulmonary toxicity has rarely been
reported. Three clinical cases of acute dyspnoea following FOLFOX therapy
(20052007) for stage III colon cancer are reported. All had an anterior resection followed by 1112 cycles of FOLFOX. Each developed rapidly progressive
dyspnoea requiring hospital admission within one week of their last cycle. One
patient required invasive ventilation in ICU. High resolution computed tomography (HRCT) showed bilateral widespread honeycomb pattern with associated ground glass opacification consistent with pulmonary fibrosis. They had
reduced lung volumes and gas transfer. Transbronchial biopsy and bronchoalveolar lavage in one patient showed an acute eosinophilic pneumonitis. Other
causes of interstitial lung disease were carefully excluded. All three patients
received high dose corticosteroids with one receiving additional cyclophosphamide. The first patient showed complete recovery following an eight week
course of corticosteroids, with resolution of the HRCT changes and improvement in lung function. The second had symptomatic improvement of dyspnoea,
but a persistent moderate reduction in gas transfer. The final patient had
persisting radiographic changes and a reduced gas transfer. He remained
dependant on ambulatory oxygen 6 months after his initial presentation. These
patients interstitial lung disease appears due to FOLFOX with oxaliplatin being
the most likely causative agent. The use of oxaliplatin chemotherapy has
increased markedly over the last 3 years and although rare, physicians should
be aware of its potential for lung toxicity. Lung function testing at baseline,
during and towards the end of oxaliplatin treatment should be undertaken and
may allow early detection and intervention in cases of pulmonary toxicity.
S PROUDMAN1, E GABBAY2, M CONRON3 on behalf of the Pulmonary

Interstitial Vascular Organisational Taskforce (PIVOT)
1
Rheumatology Unit, Royal Adelaide Hospital, Adelaide, SA, 2Advanced
Lung Disease and Pulmonary Vascular Unit, Royal Perth Hospital, Perth,
WA, and 3Respiratory Unit, St. Vincents Health, Melbourne, VIC.
The pulmonary complications of interstitial lung disease (ILD) and pulmonary
arterial hypertension (PAH) represent the leading cause of death in systemic
sclerosis (SSc).
Aims To determine the screening, diagnosis and treatment practices for ILD
and PAH of rheumatologists and respiratory physicians managing patients with
SSc.
Methods Email survey of 270 rheumatologists and 600 respiratory
physicians.
Results 42 (16%) rheumatologists and 68 (11%) respiratory physicians
responded. Of these, 63% and 59% respectively worked in a PBS PAH treatment centre. The number (mean (range)) of SSc patients seen by rheumatologists was 30 (0210) and by respiratory physicians was16 (0200). 17% and
9% of SSc patients seen by rheumatologists had ILD and PAH compared with
31% and 22%. 58% of rheumatologists screened asymptomatic SSc patients.
There was consistency in the screening tests: pulmonary function tests (PFT)
(94% rheumatologists and 97% respiratory physicians), echocardiogram
(ECHO) (79% and 79%) and CXR (67% and 77%). For suspected ILD, respiratory physicians used more bronchoalveolar lavage (26% vs 2%) and lung
biopsy (27% vs 11%). For suspected PAH, both groups used ECHO (> 90%).
More respiratory physicians used right heart catheter (51% vs 36%) and 6MWT
(62% vs 46%). In ILD, more rheumatologists used IV cyclophosphamide (CYC)
(61% vs 28%) whereas more respiratory physicians used oral CYC (42% vs
30%) and azathioprine (42% vs 27%). For PAH, more respiratory physicians
used warfarin (66% vs 40%) but there was no difference in bosentan use (62%
vs 70%).
Conclusions Over 40% of rheumatologists do not regularly screen SSc
patients. RHC is used in only 50% of patients with suspected SSc-PAH.
Supported by Australian Lung Foundation.
A65
OLIV SIG: Poster Session

TP 162
TP 161
AUDIT OF CTPA IN A REGIONAL HOSPITAL

Y RAJE, S VINCENT, G SIMPSON
Department of Thoracic Medicine, Cairns Base Hospital, Cairns, QLD 4870
Since the introduction of computerized tomographic pulmonary angiograms
(CTPA) at our institution the number of requests for this investigation at our
institution has grown at an alarming rate. The purpose of this study was to
evaluate the clinical assessment of suspected pulmonary embolism (PE).
Methods 50 CTPA were reviewed.
Results 31 female, 19 male. Mean age 50 yrs (range 2187). 26 CTPA
requests came from Department of Medicine, 21 from Emergency Department,
2 from surgical teams and 1 from oncology outpatients. 36 patients presented
with chest pain (pleuritic in 20 cases), 25 had dyspnea, 7 presented with
collapse. 4 patients had haemoptysis. Hypoxaemia was recorded in 7. None
were clinically shocked and only one had a recorded tachycardia. D-dimer
requested in 10 patients and was elevated in 9. Arterial blood gases performed
in only 10 patients (20%). 47 patients had prior chest X-ray which was normal
in 24 (48%). 8 patients had consolidation on chest X-ray, 2 pleural effusions, 2
atelectasis and 1 fractured ribs.
Recorded risk factors included 4 patients with previous DVT or PE, 4 patients
with malignancy and 6 patients were immediately post-operative.
Only 6 CTPAs (12%) demonstrated evidence of PE. Of these 2 had recent DVT
and 2 were post-operative. 1 had a history of bowel cancer. There was no
formal record of pre-test clinical probability of PE (eg Wells score) for any of the
50 cases. Retrospective calculation of the cases of PE, 4 had a Wells score of
4.5 and 1 of 4 with the remaining patient with Wells score of under 2. Only 3
patients (one with clinically probable PE) had received fractionated heparin
prior to the CTPA.
Conclusion (1) CTPAs performed at our institution have a low yield (12%).
(2) Pre-investigation clinical assessment was poor and there was poor adherence to published guidelines, (3) This results in many unnecessary CTPA
examinations generating increased work and expense for the Medical Imaging
Department and exposes many patients to unnecessary and potentially
harmful radiation exposure.
Nomination for award No.
Funding Nil.
TP 163
A COMPARISON OF THE EVALUATION AND MANAGEMENT OF

HEREDITTARY HEMORRHAGIC TELANGIECTASIA WITH
INTERNATIONAL GUIDELINES
ZD SCOUNOS, WA OLIVER, R SLAUGHTER
(Department of Thoracic Medicine) The Prince Charles Hospital, Brisbane,
QLD 4032
The evaluation and management of Hereditary Hemorrhagic Telangiectasia
involves a multidisciplinary approach according to international guidelines. The
aim of this audit was to compare the assessment process in one centre with
that of the international recommendations.
Methods Retrospective comparison was made by medical chart review of all
patients with a diagnosis of HHT between the years 1994 to 2006. Demographic along with clinical data with diagnostic investigations, complications,
treatment and genetic evaluation, including family screening was collected.
The proportion of patients evaluated and managed as per the international
recommendations was determined.
Results The audit identified 26 patients with the diagnosis of HHT, with the
mean age 58 years. Diagnostic criteria were met in 77% of the cohort. Of the
known clinical features, 54% had a family history, and 81% epistaxis. Cutaneous telangiectasia was present in 85% and visceral involvement in 92%.
Pulmonary arterio-venous malformations (PAVM) were seen in 16 patients,
cerebral AVM in 4, gastrointestinal telangiectasia was documented in 8. One
patient had a spinal (cervical) AVM, and another had pulmonary hypertension
in association with this condition. Only 8 patients underwent diagnostic or
screening investigations in accordance with the international recommendations. Furthermore, one patient was referred for a genetic evaluation.
Conclusions This clinical audit found that 31% of patients referred to this
centre were evaluated in accordance with the international recommendations.
Genetic assessment was lacking. The study supports the need for a coordinated, multidisciplinary approach to the evaluation and management of HHT in
this centre.
Paediatric SIG: Poster Session
THE PREVALENCE AND PREDICTORS OF AIRWAY

HYPER-RESPONSIVENESS IN SARCOIDOSIS
LM YOUNG1, N GOOD1, D MILNE2, W FERGUSSON1, I ZENG1, J KOLBE1,
ML WILSHER1
Departments of 1Respiratory Services and 2Radiology, Auckland District
Health Board, Auckland, New Zealand
Background While airflow limitation is the most common physiological
impairment in sarcoidosis, there are limited data on airway hyperresponsiveness (AHR). Understanding the role of AHR in sarcoidosis, if any,
may help to identify individuals who might benefit from inhaled therapies.
Aims (1) To determine the prevalence of AHR in sarcoidosis.
(2) To determine the correlation between responses to direct (using histamine)
and indirect (using hypertonic saline) bronchial challenge.
(3) To determine the clinical, physiological and radiological predictors of AHR.
Methods Subjects with a diagnosis of sarcoidosis based on typical clinical
presentation and compatible HRCT features and/or tissue biopsy and with a
baseline FEV1>35% predicted were recruited. Subjects underwent standard
hypertonic (15% fall in FEV1) and histamine (20% fall in FEV1) challenge (>1
day but <7 days apart), lung function testing and high resolution computed
tomography (HRCT) of the chest.
Results The 52 subjects (48 11 years, 35% female, 92% European, 35%
Stage I, 25% Stage II, 40% Stage III, 0% Stage IV) had well preserved lung
function overall (FEV1 = 2.8L 0.7.87% predicted). AHR was detected in 5/47
(11%) to hypertonic saline and 19/43 (44%) to histamine challenge. On univariate analysis, response to histamine challenge was predicted by conglomerate
fibrosis (p = 0.002) and reticular pattern (p = 0.05) on HRCT. The baseline %
predicted FEV1 was significantly associated with AHR on univariate
(p = 0.004), and multivariate analysis (p = 0.01) when adjusted by HRCT
patterns.
Conclusions There is a high prevalence of AHR using histamine challenge in
this study of sarcoidosis subjects. AHR most strongly associates with baseline
% predicted FEV1 but also conglomerate fibrosis and reticular pattern on
HRCT. These findings may reflect the consequence of airway remodelling
following inflammation. Further studies are warranted to confirm these findings.
Supported by Auckland Medical Research Foundation.
TP 164
NO CLINICAL BENEFIT FOR THE USE OF RESPIRATORY

SYSTEM ADMITTANCE TO MEASURE LUNG FUNCTION IN
YOUNG CHILDREN
G HALL1,3, S STRASZEK2, S STICK1,3, P SLY1,4
Respiratory Medicine, Princess Margaret Hospital, Perth, 2Institute of Public
Health, Denmark, 3School of Paediatrics & Child Health, University of
Western Australia, Perth, and 4Clinical Sciences, Institute for Child Health
Research, Perth
1
Background Upper airway shunt represents a significant source of measurement artefact in the use of the forced oscillation technique (FOT), with increasing importance in young children. Changes in respiratory system admittance,
Ars (or Zrs-1), are theoretically independent of the upper airway shunt. This
study examines the possible clinical benefit of Ars in preschool children by
assessing any increased ability to differentiate responses to bronchial challenges in the routine clinical setting. We hypothesized the use of Ars would
provide improved sensitivity to clinically relevant obstruction, bronchodilator
responsiveness (BDR) and airway hyper-responsiveness (AHR) in young children with respiratory disease.
Method Previous FOT measurements were re-analysed and Ars calculated
to derive: (1) Ars reference equations in healthy young children (n = 158); (2)
BDR in Ars, respiratory system resistance (Rrs) and reactance (Xrs) in healthy
children (n = 78), children with cystic fibrosis (n = 39), neonatal chronic lung
disease (n = 49), asthma (n = 56) and wheeze (n = 66); (3) AHR to inhaled
adenosine-5-monosphate (AMP) in 19 children. Fishers exact tests were used
to assess changes in diagnostic outcomes between Ars and conventional FOT
outcomes (Rrs and Xrs).
Results Ars was no more sensitive to bronchodilator induced changes than
conventional FOT outcomes. AMP challenges resulted in equivalent responses
measured by relative changes in Rrs and Ars while absolute changes in Ars
were the least sensitive variable.
Conclusion This study does not support a clinical advantage in using Ars in
measuring responses to either inhaled bronchodilator or AMP.
A66
TP 165
The forced oscillation technique (FOT) with broadband signals has been
employed relatively rarely in the studies on respiratory mechanics. Recent work
from our laboratory [1] indicated that the cheek support and the neck angle
have minor influence on the impedance spectra around the first antiresonance
(far,1), which makes the use of the broadband FOT especially attractive in
young children.
Methods We studied 7 healthy children (C; female: 4) and 8 children with
bronchopulmonary dysplasia (BPD; female: 3), using multiple-frequency FOT
between 8 and 256 Hz superimposed on spontaneous breathing.
Results Groups C and BPD did not differ in age (4.9 1.4 vs 6.4 1.6 yr,
NS), weight (18.9 4.7 vs 21.4 3.6 kg, NS) or height (108 11 vs
117 10 cm, NS). Total respiratory resistance was not different between the
two groups at 8 Hz (10.0 2.1 vs 9.4 2.8 cmH2O.s/l, NS) or at far,1
(16.8 0.8 vs 16.2 2.1 cmH2O.s/l, NS). In contrast, the values of far,1 were
significantly lower in the healthy children (115.9 7.7 vs 128.2 8.1 Hz,
P = 0.011).
Conclusions The subtle changes in antiresonance frequency may reflect
alterations in airway wall and lung tissue behaviour in BPD; however, further
investigations involving larger populations and detailed clinical information are
needed to address the underlying mechanisms and the clinical impact of
antiresonance.
Supported by the NH&MRC grant #404141.
Reference
1. Thamrin et al., Annals of Biomed. Eng., 2007 Oct. 18 (Epub ahead of print;
PMID: 17943446).
BS VON UNGERN-STERNBERG1, A REGLI1, F PETAK2, W HABRE1

Anaesthesia, Geneva Childrens Hospital, 1205 Geneva, Switzerland, and
2
Medical Informatics, University of Szeged, 6720 Szeged, Hungary
Lung function impairment is common in children with cardiac defects associated with increases in pulmonary blood flow/pressure. To investigate the development of bronchial hyperreactivity (BHR), an aorto-caval shunt was created in
a model of precapillary pulmonary hypertension. Surgical shunt repair was
performed to assess the reversibility of BHR.
Methods 26 rats were divided into 3 groups: Group C (n = 10) with sham
surgery, group S (n = 8) where an aorto-caval shunt was created (follow-up
4 wks), group R (n = 8) with aorto-caval shunt but surgical correction of the
shunt at 4 wks (follow-up 8 wks). In all animals, respiratory input impedance
(Zrs) was measured at baseline and following increasing doses of methacholine (Mch 2, 4, 8, 12 mcg/kg). Airway resistance (Raw), inertance, tissue
damping (G) and elastance were estimated from the Zrs spectra by model
fitting. Measurements were repeated in all animals at 4 wks and at 8 wks for
groups R and C.
Results There was a significant increase in Raw and G in group S and Rat
4 wks at baseline and following Mch (Fig.) which was reversed after surgery.
300
250
200
150
100
50
0
CONTROL
BL
4 weeks
8 weeks
SHUNT
BL
4 weeks
50
8 weeks
% Change in G
Z HANTOS1,2, GL HALL3, G NOLAN3, PD SLY2,3

Department of Medical Informatics and Engineering, University of Szeged,
Hungary, 2Telethon Institute for Child Health Research & Centre for Child
Health Research, University of Western Australia, Perth, WA, Australia, and
3
Princess Margaret Hospital for Children, Perth, WA, Australia
TP 166
PRECAPILLARY PULMONARY HYPERTENSION INDUCED BY

AORTO-CAVAL SHUNT LEADS TO A REVERSIBLE BRONCHIAL
HYPERREACTIVITY
% Change in Raw
MEASUREMENT OF BROAD-BAND FORCED OSCILLATORY

IMPEDANCE IN PRESCHOOL CHILDREN
CONTROL
BL
4 weeks
8 weeks
SHUNT
BL
4 weeks
8 weeks
40
30
20
10
0
Conclusions Inducing precapillary pulmonary hypertension by creating a left

to right shunt led to the development of BHR. The reversal observed following
surgical repair of the shunt suggests that BHR may be consequent to vascular
engorgement rather than structural changes in the airways.
Supported by a MSD scholarship of the Swiss Paediatric Respiratory Research
group and by a Swiss National Fund (SNSF 105828/1).
TP 167
IMPACT OF PREOPERATIVE PULMONARY PERFUSION ON

LUNG FUNCTION CHANGES FOLLOWING CORRECTIVE
SURGERY
BS VON UNGERN-STERNBERG1, F PETAK2, Z HANTOS2, W HABRE1
Anaesthesia, Geneva Childrens Hospital, 1205 Geneva, Switzerland, and
2
Medical Informatics, University of Szeged, Szeged, Hungary
To characterize the factors contributing to lung function impairment following

cardiopulmonary bypass (CPB), functional residual capacity (FRC), lung clearance index (LCI) and respiratory mechanics were measured in children with
pulmonary hypoperfusion (Tetralogy of Fallot, TOF n = 12) and hyperperfusion
(ventricular septal defect, VSD n = 12) undergoing surgical repair of congenital
heart disease.
Methods FRC and LCI were measured using a SF6 washout technique and
respiratory mechanics using a low frequency oscillation technique in the perioperative period.
Results While chest opening led to a significant improvement of lung
volumes and respiratory mechanics in all patients (p < 0.001), a reduction in
pulmonary blood flow during CPB decreased lung volumes and airway resistance in parallel but significantly more in children with TOF compared with
those with VSD. Re-establishing pulmonary blood flow during CPB improved
respiratory function particularly in children with TOF (Figure).
Conclusions Sternotomy had a
great impact on lung function with parallel improvement in alveolar recruitment, ventilation inhomogeneity and
airway resistance. In contrast, onset of
CPB led to lung function impairment
with a significant drop in FRC especially in children with pre-existing
hypoperfused lungs. This suggest that
pulmonary blood flow enhances alveolar stability through a tethering effect
on the alveolar walls.
Supported by a MSD scholarship of the Swiss Paediatric Respiratory Research
group and by a Swiss National Fund (SNSF 105828/1).
TP 168
LOBAR LUNG TRANSPLANTATION: A NOVEL APPROACH TO

REDUCE WAITING LIST MORTALITY IN CHILDREN REQUIRING
LUNG TRANSPLANTATION
G WESTALL1, J BURTON1, S MARESCO2, J NEGRI2,3, M BUCKLAND4,
C ROBERTSON3, G SNELL1
1
Lung Transplant Unit, 2Department of Cardiothoracic Surgery and
3
Department of Anaesthesia, Monash University Medical School, Alfred
Hospital, Melbourne, Vic 3181, and 4Department of Respiratory Medicine,
Royal Childrens Hospital, Melbourne, Vic 3052
Children with advanced lung disease being considered for lung transplantation
are likely to spend disproportionately longer periods on transplant waiting lists
before appropriately sized donor organs become available. These longer
waiting times reflect the lower organ donation rates seen in children; rates that
are significantly lower than those reported in the adult population. We describe
two children with advanced lung disease who deteriorated whilst on the waiting
list for lung transplantation, and in the absence of appropriately sized donor
lungs, underwent lobar lung transplantation.
Methods We describe the clinical course of two children, aged 9 and 13
years old, with advanced lung disease secondary to post-mycoplasma obliterative bronchiolitis and cystic fibrosis-associated bronchiectasis, respectively.
Results Both children received a cutdown bilateral lobar transplant from
two oversized adult brain-dead organ donors. In both cases the transplant
operation involved implantation of the right middle and upper lobes, and of the
left upper lobe from the donor.
Conclusion Given the low organ donation rates in children, and in the
absence of appropriately sized donor lungs, novel strategies such as lobar
transplantation must be considered, particularly when children continue to
clinically deteriorate whilst on the lung transplant waiting list.
TP 169
A67
TP 170
MATERNAL FACTORS DO NOT PREDICT EMPHYSEMA

SEVERITY IN YOUNG ADULT SURVIVORS OF
BRONCHOPULMONARY DYSPLASIA
A RANDOMISED CONTROLLED STUDY ON EDUCATION

INTERVENTION FOR CHILDHOOD ASTHMA BY INDIGENOUS
HEALTH WORKERS IN THE TORRES STRAIT
DC CHAMBERS1, CP MURRAY2, J LOUW2, J NEWNHAM3, N FRENCH3,

A WILSON1,4
1
Royal Perth Hospital, Perth, Western Australia; 2Department of Diagnostic
Imaging, Royal Perth Hospital; 3Neonatal Clinical Care Unit, King Edward
Memorial Hospital, Perth. 4Princess Margaret Hospital, Perth
PC VALERY1,2, IB MASTERS3, B TAYLOR4, P OROURKE1,2, Y LAIFOO5,

AB CHANG3,6
1
Queensland Institute of Medical Research, 2Australian Centre for
International and Tropical Health and Nutrition, 3Royal Childrens Hospital,
4
The Asthma Foundation of Queensland, 5Thursday Island Primary Health
Care Centre, and 6Menzies School of Health Research
Data from the West Australian adult outcomes of extreme preterm birth study
suggest that adult survivors of bronchopulmonary dysplasia (BPD) may be left
with functional and structural pulmonary abnormalities, most notably emphysema. Animal data suggest that the antenatal administration of corticosteroids
may adversely affect lung development. We therefore sought to determine if
maternal variables, including administration of corticosteroid, could predict
emphysema severity in adulthood.
Methods BPD subjects (birthweight < 1500 g and oxygen dependence at 36
weeks post-menstrual age) born prior to 1988 were identified and recruited
prospectively via the statewide neonatal follow up program as previously
described. Pulmonary function tests and thin selective inspiratory and expiratory computerised (CT) images were acquired and scored for emphysema
severity (voxel index (%)). The obstetric history was obtained from retrospective review of case notes.
Results 21 adults (12 females, aged 1834) were studied, 2 declined CT. All
subjects had abnormal CT findings. Fifteen (79%) had areas of emphysema.
Emphysema score and FEV1 were not influenced by the administration of
antenatal corticosteroids, indication for delivery, maternal age or presence or
absence of chorioamnionitis.
Conclusion Maternal factors, including the administration of antenatal corticosteroids, do not predict the long term respiratory outcome of BPD. The
factors determining the severity of emphysema in this group remain unknown.
The prevalence of childhood asthma is high in the Torres Strait. Children have
generally more severe asthma and asthma knowledge is poor. However, there
is no culturally appropriate asthma education program for these children. We
are conducting a randomized controlled trial to examine the additional benefits
of an education intervention by Indigenous Health Care Workers (HCW) on
asthma outcomes. We describe the studys objectives, design and baseline
measurements.
Methods Children with wheeze were reviewed by two paediatric respiratory
physicians using a standardized protocol; children with asthma were eligible.
After obtaining informed consent children were randomly allocated to: (1) three
additional asthma education sessions with a HCW; or (2) no additional education from a HCW. Trained HCWs carried out the education sessions using
culturally appropriate tools. Primary outcome was the number of unscheduled
hospital/doctor visits due to asthma exacerbation. All children were
re-assessed at 12 months.
Results We enrolled 113 children aged 1 to 17 years, 81% were Torres Strait
Islanders and 12% Aboriginal and Torres Strait Islanders. The clinical spectrum
of asthma was: 51% infrequent episodic asthma, 22% frequent episodic
asthma and 27% chronic asthma. Eighteen percent of the children knew what
a written Asthma Action Plan was; 8.5% had one. Carers assessment of
knowledge of medications showed that 52% could not name any asthma
medication used by their child, 40% could not explain dosage, and 67% could
not explain how beta2 agonists worked.
Conclusions Asthma knowledge and possession of asthma action plans in
this cohort is poor at baseline. There is substantial room for improvement and
additional asthma education by HCWs potentially has significant benefits.
TP 172
SPIRO-GPA TRIAL OF SPIROMETRY IN GENERAL PRACTICE
Primary Care SIG: Poster Session

TP 171
AUSTRALIAN PREDICTIVE EQUATIONS FOR THE IMPULSE

OSCILLOMETRY SYSTEM
W NEWBURY1,3, A CROCKETT2, J NEWBURY3
Menzies School of Health Research, Royal Darwin Hospital Campus,
Casuarina, NT 0810, 2Primary Care Respiratory Unit, Discipline of General
Practice, University of Adelaide, SA 5005, and 3Spencer Gulf Rural Health
School, Universities of Adelaide and South Australia, SA 5005
Impulse Oscillometry System (IOS) measures respiratory function during

normal breathing by transmitting mixed frequency rectangular pressure
impulses down the airways and measuring reflected pressure. Computer
analysis calculates respiratory impedance and its components, airways resistance and reactance, at a range of frequencies from 0.1 Hz to 150 Hz. No
previous Australian normative data exists. The IOS software generates predictive normal values for each of the parameters measured including total airway
resistance (R5), the proximal airway resistance (R20) as well as peripheral
capacitive reactance (X5). However, they are based on German data.
Methods Cross-sectional study of 100 community dwelling adults, with 10
males and females per 10-year cohort. Inclusion criteria: age range 2574
years, apparently good respiratory health. Exclusion criteria: smokers, asthmatics and others with acute or chronic respiratory disease. Both IOS and
spirometry were conducted on all participants.
Results Australian predictive normal equations have been generated and
compared to the current published equations. The IOS parameters have been
correlated with the spirometric data. Results have been analysed by gender,
age, height and weight and compared with the predictive normal values for
each parameter provided by the German manufacturer of the IOS instrument.
Analysis includes calculation of mean range, and lower limit of normal.
Conclusions A preliminary set of Australian predictive equations have now
been produced for the IOS. These have been compared with international
equations. IOS has potential application in a range of respiratory disease
states and in population screening for occupational health (e.g. mining, & high
dust load environments).
Supported by PHC RED.
MJ ABRAMSON1, R SCHATTNER1, FCK THIEN2, ND SULAIMAN3,

E DEL COLLE4, R ARONI5
1
Department of Epidemiology & Preventive Medicine, Monash University,
2
AIRmed, The Alfred, Melbourne, Vic 3004, 3Department of General
Practice, University of Melbourne, Vic 3010, and 4Pulmetrics P/L, East
Doncaster Vic 3109, and 5Monash Institute of Health Services Research,
Clayton, Vic 3168
Rationale Although clinical practice guidelines for both asthma and COPD
recommend spirometry for diagnosis and monitoring, beneficial effects on the
management of chronic respiratory diseases in general practice have not been
established. We hypothesized that spirometry would improve health outcomes
compared to usual care.
Methods We are conducting a single masked RCT with 3 arms: Group A
receive 3 monthly spirometry and followup, Group B receive spirometry before
and after the trial and Group C usual care. 45 general practices were recruited
though Divisions of General Practice in Melbourne. Invitations were mailed by
31 of these practices to patients who had been prescribed inhaled medications
during the previous 6 months. Participants returned respiratory and generic
quality of life questionnaires and an asthma score card. Groups A and B were
tested on a MicroMedical turbine spirometer following ATS/ERS guidelines.
Results 351 eligible patients (275 adults, 50 children aged 813 and 26
youths aged 1417 years) entered the trial. 122 were randomized to Group A,
134 to Group B and 95 to Group C. The mean (SD) age of adult participants
was 54.3 (12.7), children 10.3 (1.7) and youths 15 (1.1) years. There were 130
males and 221 females. The adults were highly symptomatic in the previous 12
months: 82% reporting wheeze, 50% chest tightness on waking, 74% shortness of breath on exertion, 61% nocturnal cough, 46% morning cough and
75% sputum. Symptoms of chronic bronchitis were reported by 39% of adults
and a diagnosis of COPD by 19%. Asthma was reported by 84%, confirmed by
a doctor in 96% and 55% had experienced an attack in the last 12 months. Only
35% had a written asthma action plan. 37% of adults had ever visited a hospital
ED and 28% had been admitted.
Conclusion It is possible to recruit asthma and COPD patients from general
practice and to randomize them to spirometry or usual care. Whether spirometry is associated with fewer symptoms, changes in medication, uptake of
action plans or improvement in lung function or quality of life requires further
followup.
Supported by NHMRC.
A68
TP 173
STRATEGIES USED IN RECRUITMENT FOR A RCT IN PRIMARY

CARE
S SHAH1, JK ROYDHOUSE1, B TOELLE2, S SAWYER3, C JENKINS2 for
the PACE Australia Management Committee
1
University of Sydney, 2Woolcock Institute of Medical Research, Sydney,
NSW 2006, and 3Royal Childrens Hospital, Melbourne, VIC 3052
Respiratory Infectious Diseases SIG:

Poster Session
TP 174
CLINICAL PHENOTYPES OF BRONCHIECTASIS

P KING1,2, M FARMER1, P HOLMES1
Respiratory Medicine Monash Medical Centre/Dandenong Hospital, and
2
Monash University Department of Medicine, Clayton, VIC 3168
It is widely held that recruitment of general practitioners for research can be

challenging. In this paper, we discuss the recruitment experience from a current
study evaluating the impact of an educational asthma intervention on patient
outcomes. Our aim is to describe the two different strategies utilized to date: (1)
in-house through an academic department of GP and (2) outsourced to a
private GP organization.
Methods Initial interest was generated through faxes, presentations at GP
Divisional meetings and newsletter advertisements. GPs who expressed interest were visited by project staff to discuss the study further. A major difference
was recruiting one GP per practice in the first strategy versus multiple GPs per
practice in the second strategy. To assess the strategies, we examined participant characteristics, number of GPs recruited and number retained.
Results Participant characteristics: Under both strategies, 30% of recruits
had trained in Asia and 54% were women. The first strategy recruited more
GPs who spoke at least two languages at home (85% vs 42%) and the second
strategy recruited more recently graduated GPs (58% vs 50%). Recruitment:
The first strategy recruited 35 GPs over 6 months and the second recruited 34
GPs over 3 months. Retention: 19 GPs (54%) from the first strategy stayed in,
compared to 29 (85%) from the second.
Conclusions Whilst absolute numbers of GPs recruited were similar, retention was much higher under the second strategy. Recruitment in primary care
is difficult and requires a range of approaches which need to be re-evaluated
and adapted as necessary during the course of the study.
Supported by the Australian Government Department of Health and Ageing.
Bronchiectasis is a heterogeneous condition with a large number of causative

factors and range of symptoms. The classification of this condition is often
confusing and hard to remember. The aim of this study was to classify non-CF
bronchiectasis into different clinical phenotypes.
Methods 178 consecutive patients with non-CF bronchiectasis confirmed on
high resolution CT scanning had a detailed clinical, spirometric and laboratory
assessment performed by a respiratory physician (PK/MF/PW) and were then
followed up for an average of 9 4 years (mean and SD) for a total of over
2000 reviews.
Results 160 of the 178 patients (90%) could be classified as belonging to 3
phenotypic groups; 1) bronchiectasis arising in childhood, 2) bronchiectasis
occurring in smokers and 3) bronchiectasis occurring in the elderly. Each group
had different features which are listed in the table (results are mean & SD).
Phenotype of bronchiectasis
Childhood
Smokers
Elderly
Number of subjects and %

Age of onset of cough (yrs)
Duration of productive cough
(yrs)
Dyspnoea (MRC scale)
Sputum volume (mls)
FEV1 (% predicted)
FVC (% predicted)
98 (55%)
65
47 23
43 (24%)
52 10
10 7
19 (11%)
73 6
32
1.8 1
42 31
70 22
87 18
2.9 1.4
21 15
58 25
78 27
1.5 1.1
12 9
86 10
95 12
Conclusion Most subjects in this cohort of non-CF bronchiectasis patients

could be classified as belonging to 3 distinct phenotypic groups; bronchiectasis
arising in childhood, smokers and elderly.
TP 175
RAPID ANTIGEN TESTING (RAT) TO DIAGNOSE INFLUENZA IN

A HOSPITAL SETTING
BENJAMIN CH KWAN1, CAROL EMERSON2, SUZANNE RYAN2, PAUL S
THOMAS1, KATE CLEZY2
1
Department of Respiratory Medicine, Prince of Wales Hospital. Sydney,
NSW 2031, and 2Department of Infectious Diseases, Prince of Wales
Hospital, Sydney, NSW 2031
Influenza A or B can cause an acute respiratory illness, mainly during the winter
months. In the community the diagnosis of influenza can be made using clinical
criteria once an outbreak is established; whereas in hospital an accurate
diagnosis is essential for infection control measures. Rapid viral diagnostic
tests, e.g. immunofluorescence (IF) assays, enzyme immunoassays (EIA), and
polymerase chain reaction (PCR)-based tests may be appropriate alternatives
to the gold standard of laboratory viral cultures. Point of care RAT allows
rational use of antiviral therapy both for patient treatment and prophylaxis of
health care workers and family contacts.
Methods During the winter months July to September 2007, subjects were
recruited on the basis of fever with at least one respiratory and constitutional
symptom. In addition to standard clinical evaluation and chest x-ray, an anterior
nasal swap was collected. Using the QuickVue A+B test, a RAT kit using
monoclonal antibodies specific for influenza A and B virus antigens, a result
was available within 15 minutes.
Results We recruited 125 subjects through emergency, respiratory and infectious diseases departments. Of these, 12 results returned positive, with 11
being Influenza A and 1 being Influenza B. RAT costs $30 per testing, compared with $150 for a laboratory EIA/viral culture.
Conclusions Previous standard of care was for laboratory EIA with turnover
time of 2472 hours. Our rapid results allowed prompt clinical decisions and
early intervention such as antiviral therapy for patient and contacts and isolation of cases. It may also prove to be cost effective.
Supported in part by BioCryst pharmaceuticals, Inc.
TP 176
RECURRENCE OF CULTURE POSITIVE TUBERCULOSIS IN NEW

SOUTH WALES
CC DOBLER, ABH CRAWFORD, GB MARKS
Department of Respiratory Medicine, Liverpool Hospital, Sydney, NSW 2170
There are few data on the long term outcomes of treatment for tuberculosis
(TB) by directly observed therapy (DOT) in low-incidence settings. The aim of
this study was to assess the incidence of recurrent TB in NSW.
Methods Data linkage was performed within the NSW Department of Health
TB notifications database to identify cases that had more than one TB notification between 1994 and 2006. Recurrent tuberculosis was defined to include
all patients with two or more culture positive episodes at least 6 months apart,
where patients had received at least six months treatment for the initial
episode. In cases where data contained within the notification details was not
sufficient to allow us to distinguish between true cases of recurrent disease,
duplication notification for the same episode or persistent disease after incomplete treatment, additional information was obtained from the Area TB
coordinator.
Results There were 5723 TB notifications between 1994 and 2006 with 3731
being culture positive. 15 cases of recurrent culture positive disease after
completed treatment for the first episode were identified (recurrence rate:
0.4%).
Conclusions In a population with a low TB incidence, treatment of active
tuberculosis with DOT results in a very low rate of disease recurrence over a
long period of follow-up.
Support NHMRC CCRE in Respiratory and Sleep Medicine.
TP 177
A69
TP 178
NUCLEOCYTOPLASMIC LOCALIZATION OF RHINOVIRUS 3C

PROTEASE
CHRONIC NECROTIZING PULMONARY ASPERGILLOSIS

RE-VISITED
D LI, H DAGHER, M LINDSAY, D JANS, P BARDIN, R GHILDYAL

Department of Respiratory Medicine, Monash University and Medical Center
and Department of Biochemistry and Molecular Biology, Monash University,
Clayton, Vic 3800
V AIYAPPAN, A SARACINO, J HO, M HOWARD, P DANBY

Department of Medicine, Maroondah Hospital, Ringwood East, VIC 3135
Introduction Rhinoviruses (RVs) are the major cause of viral-induced exacerbation of asthma. To date, the molecular mechanisms of RV pathogenesis
are not understood. Recent findings suggest that RV pathology may involve
host cell nucleocytoplasmic trafficking, inhibiting key cell functions such as
transcription and translation. The study aims to investigate the mechanism of
RV 3C protease nuclear trafficking.
Methods HeLa cells were infected with RV or transfected with plasmids and
cellular localization of 3C analysed at various times thereafter using immunofluorescent confocal microscopy and Western blotting with specific antibodies.
Results 3C protease was predominantly present in nuclei of RV infected cells
up to 6 hours after infection, becoming increasingly cytoplasmic thereafter. The
nuclear membrane of infected cells became progressively indistinct with time.
Using a specific inhibitor we also found that 3C utilizes the CRM-1 nuclear
export pathway. 3C was predominantly in the form of 3CD in both cytoplasm
and nucleus of infected cells; mature 3C protease was also detected from 6
hours after infection. Deletion analysis indicats that the nuclear localization
domain and a nuclear export signal are most likely to be present within the N
terminal 64 amino acids. The nuclear export signal is inhibited in the full length
protein, via an unknown mechanism.
Conclusion Our data suggest that 3C and 3CD proteins localize to the
nucleus in infected cells where they may play a key role in RV pathogenesis by
disrupting cellular transcription and the nuclear transport machinery.
Chronic necrotizing pulmonary aspergillosis (CNPA) is a relatively uncommon,

sub-acute, locally destructive process due to Aspergillus invasion of the lung.
The incidence and prognosis of CNPA are poorly described.
Case report We present a case of CNPA in a patient on intermittent low dose
steroid therapy and recurrent refractory exacerbations of chronic obstructive
pulmonary disease (COPD).The patient presented with worsening shortness of
breath and productive cough requiring recurrent inpatient admissions. Following multiple Aspergillus positive sputum cultures, a suspicious chest CT and
review of the current literature, CNPA was suspected and the patient underwent bronchoscopy. Positive bronchial washings, raised serum IgE and
strongly positive serum Aspergillus precipitins eventually lead to the definitive
diagnosis of CNPA.
Progress The patient meanwhile developed MRSA pneumonia and septicemia (confirmed by positive blood and sputum cultures) and had to be intubated
for severe respiratory failure. She was treated with antibiotics and voriconazole. She made a remarkable recovery, was extubated and is now recuperating. She is currently on Voriconazole and further imaging has shown good
radiographic resolution.
Discussion This patients risk factors, clinical presentation and delayed and
difficult diagnosis raise many questions and possible failures in current clinical
approaches to CNPA.
TP 179
DISTRIBUTION OF SAa2,6GAL and SAa2,3GAL LINKED

RECEPTORS IN HUMAN RESPIRATORY TRACT AND
INFLUENZA VIRUS REPLICATION
A HSU, T GRISSELL, PAB WARK
Department of Respiratory and Sleep Medicine John Hunter Hospital and
the School of Medicine and Public Health, University of Newcastle, NSW
2305, Australia
Human influenza virus is found to bind preferentially to SAa2,6GAL receptors
found in the upper respiratory tract, while avian viruses bind to SAa2,3GAL
receptors expressed in lower airways. This is thought to affect the ability of
transmission to humans. Our aim was to study the ability of avian and human
influenza strains to infect bronchial epithelial cells and relate this to levels of the
sialic acid receptor expression.
Methods Calu-3 cells were used as a proximal airway cell and A549 were
used as distal airway cell. Human primary bronchial epithelial cells (pBECs)
were obtained from healthy, asthmatic, and COPD volunteers by endobronchial
brushing.
Epithelial cells were stained with Sambucus nigra lectin that binds SAa2,6Gal
receptor, and maackia amurensis lectin II that binds to SAa2,3Gal. The cells
was analysed by flow cytometry. Human influenza A/H3N2/Wellington strain
and low pathogenic avian influenza A/H11N9/Sandpiper were chosen and were
used at an MOI of 0.005 to infect cells. The supernatants were harvested at
48 hr post infection, of which was then analysed by plaque assay for virus
replication.
Results The Calu-3 showed greater expression of SAa2,6Gal linkage than
SAa2,3Gal linkage, and A549 displayed slightly higher expression of both
receptors compared to pBECs. Despite this human and avian influenza virus
replicated to similar titre at 15,000 pfu/ml in both cell lines, but showed low
replication in pBECs.
Conclusion Sialic acid receptor expression alone does not account for the
ability of influenza virus to infect and replicate in host cell. Intrinsic innate host
cell immunity also limits virus replication in pBECs. Supported by the Centre of
Asthma and Respiratory Disease.
TP 180
QUANTITATIVE REAL-TIME PCR FOR PNEUMOCOCCUS IN

PATIENTS WITH PNEUMONIA
G WATERER1, C KEE1, S PALLADINO1, I KAY1, T PRYCE1, T LISBOA2,
J RELLO2
1
Royal Perth Hospital, and 2Joan XXIII University Hospital, Tarragona, Spain
Background Treatment of community-acquired pneumonia remains based
on best guess empiric algorithms because of the poor utility of current pathogen tests. Furthermore our ability to stratify patients into risk groups is crude at
best, relying on scores such as the pneumonia severity index or the CURB-65
have major limitations. We have been slowly improving real-time PCR assays
for pneumococcus as a clinical tool in patients with pneumonia.
Methods Building on previous research we assesed two targets in the autolysin (lytA) gene and the pneumolysin (ply) gene of S.pneumoniae using the
LightCycler instrument and Fluorescence Resonance Energy Transfer (FRET)
probes. All common S. pneumoniae serotypes were detected while other
bacteria and viruses were not. The lytA target had the best sensitivity with a
detection range between 21 ng to 21 fg. Both assays were then applied to
whole blood samples from 400 adult patients with community-acquired pneumonia, all of whom had blood cultures prior to antibiotic administration and
urinary antigen testing for S.pneumoniae.
Results The lytA PCR had the best performance characteristics with a sensitivity more than twice that of blood cultures in the clinical samples. Most
PCR+ve/culture ve patients had positive urinary antigen tests. There was
clinical evidence that urinary antigen +ve/ PCR ve patients were false +ves.
Most significantly there was a strong correlation between quantitative bacterial
count and clinical outcome.
Conclusions Real-time quantitative PCR for pneumococcus has significant
potential as both a diagnostic and therapeutic tool in patients with pneumonia.
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TP 181
TP 182
TUBERCULOSIS PREVALENCE IN ANANGU PITJANTJATJARA

YANKUNYTJATJARA LANDS IN SOUTH AUSTRALIA
PATTERNS OF ANTIBIOTIC PRESCRIPTION FOR INPATIENT

TREATMENT OF COMMUNITY-ACQUIRED PNEUMONIA
A RETROSPECTIVE AUDIT
G FOX1, K GELL2, J REED3, R STAPLEDON3, R SCHULTZ4, P TORZILLO2

Department of Respiratory Medicine, Royal Prince Alfred Hospital, NSW
2050, 2Nganampa Health Council, South Australia, 3South Australian TB
Service, Royal Adelaide Hospital, Adelaide, SA 5000, and 4Centre for
Disease Control, Alice Springs, NT 0870
M HAYES, JR WHEATLEY, TD ROBINSON

Department of Respiratory and Sleep Medicine, Westmead Hospital, NSW
2145
The Pitjantjatjara Lands are situated in the north-western corner of South

Australia, occupying an area of over 120 000 square kilometres with a population of approximately 3000. The population lives in small communities or
homelands, and there is a high level of mobility between this region and other
Aboriginal communities in South Australia and the Northern Territory. Nganampa Health Council provides all health care services to the region. Specialized support for TB control comes from both the South Australia TB service
based at Royal Adelaide Hospital as well as a Centre for Disease Control in
Alice Springs.
The prevalence of tuberculosis (TB) in this predominantly indigenous community is thought to be significantly higher than the national rate. There are
considerable challenges in detecting and managing tuberculosis, relating to the
communitys geographical remoteness, migration of populations and access to
health services.
The aims of this study are to quantify the prevalence of tuberculosis in the
Pitjantjatjara Lands, and describe the significant barriers to TB diagnosis and
treatment.
Methods A retrospective study of all diagnoses of tuberculosis within the
Pitjantjatjara lands in the period 19952006. Outcomes include measures of
tuberculosis diagnosis, the rates of completed TB treatment and rates of
tuberculosis drug resistance.
The study will draw conclusions about the reasons for high levels of TB
prevalence in this community and identify barriers to effective tuberculosis
treatment.
Conflict of Interest NO.
TP 184
Sleep & Physiology SIG: Poster Session

TP 183
TIDAL BREATHING TEST WITH ULTRASONIC FLOW SENSOR

DETECTS AIRWAY OBSTRUCTION
1
Patients admitted to hospital with a diagnosis of community-acquired pneumonia (CAP) are usually treated with intravenous (iv) antibiotics irrespective of
pneumonia severity. Available guidelines vary in recommended timing and
indications for switching to oral antibiotics.
Aim To examine the patterns of antibiotic choice and delivery method (iv, oral
and time to switch) in patients admitted with CAP.
Methods A retrospective chart review of admissions to the Respiratory Unit
over a 12-month period with a Diagnostic-Related Group (DRG) coding of
pneumonia. 41 charts were reviewed. Data collected included patient demographics, clinical features at presentation (temperature, pulse rate, respiratory
rate, BP, oxygenation), initial investigations, initial antibiotic regime, time
to change (iv to oral), subsequent antibiotic regime and duration, time to
defervescence, length of stay and outcome. Pneumonia severity was
calculated using the revised British Thoracic Society system (CURB-65),
score 2 = severe.
Results 3 patients were excluded due to incorrect coding. Of the 38 patients,
age was 50 21 (mean SD) yrs and 25 (66%) were male. 28 patients (74%)
were febrile at presentation and the median CURB-65 score was 1 (range 04).
37 patients (97%) received iv antibiotics. The CURB-65 score was 0 or 1
(non-severe) in 25 patients and 22 of these patients received a combination of
iv ceftriaxone and a macrolide. Time to defervescence was 2.9 2.3 days.
Time from defervescence to switching to oral therapy was 3.4 2.8 days. In
non-febrile patients, time to switch was 4.74.3 days. Length of stay was
8.713.0 days.
Conclusions The time between defervescence and switch to an oral regime
was relatively long, possibly contributing to an increased length of stay. Many
patients received ceftriaxone even with a CURB-65 severity rating of 0 or 1.
Implementing local guideline-based treatment protocols may reduce length of
stay.
L BECKERT , C BUESS , H MAGNUSSEN

Department of Respiratory Medicine, Christchurch Hospita, Christchurch,
New Zealand, 2ndd Medizintechnik, Technopark, Zrich, Switzerland, and
3
Pulmonary Research Institute, Grosshansdorf, Germany
Ultrasonic flow sensors can determine flow, volume and molar mass (MM) of
the gas flow simultaneously. During tidal breathing the expired molar mass
curve can be used to compute CO2 over expired volume and a Capnography
Index (CPI) can be computed. The relationship between CPI and COPD
classification according to GOLD was investigated.
Methods Prospective, controlled trial. Consecutive patients who underwent
routine lung function were enrolled to participate in a tidal breathing test using
an ultrasonic flow sensor. Each test consisted of three tidal breathing recordings of 60 sec. Flow, volume and molar mass were measured at 200 Hz and
data were acquired using prototype WBreath data acquisition software. Mean
expirograms (MM over volume) were computed and the measurements were
analyzed to determine the slope of exhaled phase II (S2), the slope of phase
III (S3) and the relationship between S2 and S3 (CPI = S3/S2). GOLD stages
were determined from the lung function results and the ERS predicted values.
Results 53 volunteers participated in the study with a mean age of 62 (SD
14), 23 were male, mean BMI 26 (SD 5), 17 had never smoked. The mean
pack/year smoking history was 38. There was a clear relationship between
GOLD stage and CPI: GOLD stage normal had a mean CPI of 5.5 (SD 3.7,
n = 21), stage severe had a mean CPI of 13.7(SD = 3.9, n = 7).
Conclusion Computation of CPI based on tidal breathing analysis using an
ultrasonic flow and MM sensor correlates well with GOLD stages. It may
therefore be possible to use a simple tidal breathing test to determine the
severity of airways disease.
SLEEP HEALTH IN TETRAPLEGIA THE FEASIBILITY OF

CONTINUOUS POSITIVE AIRWAY PRESSURE (CPAP)
TREATMENT FOR OBSTRUCTIVE SLEEP APNOEA (OSA) IN
ACUTE TETRAPLEGIA
DJ BERLOWITZ1, J ROSS12, J SPONG1, D RILEY1, RJ PIERCE1,
DJ BROWN2
1
Institute for Breathing and Sleep, and 2Victorian Spinal Cord Service, Austin
Health, Victoria 3084
Background OSA is common in tetraplegia and appears within weeks of
injury. Although CPAP treatment is efficacious in able-bodied subjects, case
series suggest that CPAP is poorly tolerated in tetraplegia. No prospective
study has examined CPAP efficacy or adherence in tetraplegia.
Aim To determine the feasibility of CPAP use to treat OSA following acute
tetraplegia.
Methods All acute admissions who consented and fulfilled the inclusion and
exclusion criteria underwent full, portable polysomnography. Those found to
have an apnoea hypopnoea index of >10 events per hour (OSA) were offered
CPAP, delivered via an auto-titrating device.
Results To date, 25 patients have been admitted (11 excluded, 3 refused
consent). No significant, adverse events have been observed. Two patients did
not have OSA. Of the nine with OSA, four are mid-study, two had incomplete
follow-up (1 returned to UK and 1 refused 3 month assessment), two adhered
with CPAP and one did not due to severe, pre-existing nasal obstruction.
Preliminary analyses suggest that those who adhered to CPAP had a marked
reduction (80% compared with 1040%) in sleepiness and a greater reduction
in the functional outcomes of sleepiness compared to either those without OSA
or who were unable to use CPAP. Patient accrual, recruitment and completion
rates are consistent with our initial estimates. Study recruitment will be completed by end-October 2007.
Conclusion Initial data suggest that auto-titrating CPAP is a feasible treatment for OSA in acute tetraplegia. These data will be used to finalize planning
for a multi-national, multi-centre randomized controlled of therapy.
This research was supported by the Transport Accident Commission.
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TP 185
ABSTRACT WITHDRAWN
TP 186
ARTERIALISED VENOUS SAMPLING AN ALTERNATIVE TO

ARTERIAL BLOOD GASES IN CHRONIC HYPERCAPNIC
RESPIRATORY FAILURE?
C HOLLIER1,2, C MENADUE1,2, D FLUNT1,2, AJ PIPER1,2
Department of Respiratory and Sleep Medicine, Royal Prince Alfred
Hospital, NSW 2050, and 2Woolcock Institute of Medical Research, NSW
2050
Serial measurement of arterial carbon dioxide (PaCO2), pH and bicarbonate

(HCO3) is essential in the management of patients with hypercapnic respiratory failure (HRF). This information is usually obtained from a sample of arterial
blood (ABG). The procedure can be painful and distressing for patients, and is
sometimes technically difficult due to obesity or contractures. Our aim was to
determine the validity and feasibility of arterialized venous blood (AV) sampling
as an alternative to ABGs in measuring PaCO2, pH and HCO3- levels in patients
with chronic HRF.
Method Eighteen patients completed the study. Venous blood was arterialized by heating forearm skin to a temperature of 4245C with an electric
heating pad. An AV sample was taken from a cannula positioned in a vein of the
heated forearm simultaneously with an ABG. In addition, the reliability of AV
sampling within the recommended temperature range (4245C) was investigated in ten healthy volunteers placed on volume cycled ventilation in order to
maintain constant ventilation. AV samples were taken at 0.5C temperature
intervals from 42.545C
Results The table below summarizes results for validation of AV sampling:
Patients n = 18
ABG (mean SD)
AV Sample
Mean diff (95%CI)
ICC
PaCO2 (mmHg)
pH
HCO3-
49.1 12.0
7.40 0.03
29.1 5.1
47.9 7.6
7.38 0.03
27.7 3.3
1.2 (-2.04.3)
0.01 (00.02)
1.4 (0.22.7)
0.77
0.86
0.80
In healthy subjects, varying the temperature from 42.545C did not affect
CO2, pH or HCO3. Repeated measures of CO2, pH and HCO3- were highly
consistent [intraclass correlation coefficient (ICC(C,1)) 0.99, 0.97 and 0.92
respectively].
Conclusions Arterialized venous sampling was well tolerated and produced
valid measures of arterial CO2, pH and HCO3-. Determination of test-retest
reliability is required to establish AV sampling as an alternative to ABGs in
patients with chronic HRF. The role of AV sampling in acute HRF is yet to be
established.
TP 187
IDENTIFICATION OF ONSET AND TERMINATION OF

INDIVIDUAL SLEEP APNEA EVENTS FROM SINGLE LEAD ECG
RECORDINGS
C KARMAKAR, A KHANDOKER, M PALANISWAMI
Dept. of Electrical and Electronic Engineering, Melbourne University,
VIC 3010
Based on the evidence that cardiovascular dynamics are altered due to
obstructive sleep apnea, this study aims to identify the onset and termination of
each apnea event using power spectral density (PSD) and morphological
features of single lead ECG signal over 5 second period.
Methods ECGs from 4 patients overnight sleep studies were examined for
location of the pre-scored apnea events. Onset (n = 1995), maximum
(n = 6751) and termination (n = 1996) of each apnea event and normal events
(n = 11219) were annotated on 5 second windows. Features extracted were
PSD, amplitudes of R and T wave of 5 second ECGs. Receiver operating
Characteristics (ROC) analysis was used to gauge the event recognition ability
of all features.
Results The maximum areas under ROC curves for PSD values of ECG
during normal/onset, normal/maximum and termination/normal were 0.82 (frequency range: 5260 Hz), 0.86 (range: 4577 Hz) and 0.83 (range: 5162 Hz)
respectively. On the other hand, maximum ROC areas of mean amplitudes of
R and T wave for the same events were found to be 0.76, 0.88 and 0.80
respectively.
Conclusions The features PSDs (5162 Hz) and mean amplitude of T
waves over 5 seconds ECG signal turned out to have valuable information for
identification of onset and termination of an apnea event. The results from this
study will be useful in estimating correct apnea-hypopnea index (AHI) based
solely on shorter segments (only 5 seconds) of ECG signals.
Supported by the ARC Linkage grant.
TP 188
USE OF A COMPREHENSIVE SCREENING PROTOCOL IN

IDENTIFYING OSA PATIENTS FOR THE VERY LOW CALORIC
DIETA PILOT STUDY
ANNA TAI1, TAM EATON1, RINKI MURPHY2, WARWICK BAGG2
Respiratory Department, Auckland City Hospital, Auckland, and
2
Endocrinology Department, Auckland City Hospital, Auckland
Weight loss causes an improvement in the severity of OSA, however substantial weight loss is very difficult for obese patients. The Very Low Caloric Diet
(VLCD) has been shown to be successful in causing significant weight loss in
obese patients. This is a pilot study on the use of a formal screening protocol
to identify OSA patients who are potentially eligible for the supervised VLCD
program offered by the Endocrinology Department at Auckland City Hospital.
Method 344 consecutive patients who attended the sleep laboratory at ACH
between June to December 2006 were screened using the protocol. Patients
who are eligible to be considered for the VLCD program are identified as having
a combination of obesity (BMI > 30), OSA (AHI > 5 on sleep study) and being
residents within the Auckland District Healthboard region.
Results 243/ 344 patients screened did not fulfil the inclusion criteria: 171
lived outside the ADHB region; 71 had BMI < 30; 7 patients did not have OSA
(AHI < 5). 101 patients fulfilled the inclusion criteria. 54/101 patients (54%)
were excluded due to medical or psychiatric contraindications to VLCD.47
patients (47%) who did not have contraindications to VLCD were contacted. 33
patients were contacted successfully. 14 patients were either unavailable to
phone contacts on 3 separate days or were disconnected. 12/101 patients
consented to being referred (12%). 21/101 patients declined referral (21%).
Conclusion This pilot study is the first study using a formal comprehensive
screening protocol in the recruitment of obese OSA patients into a medically
supervised VLCD program. Only a small proportion (12%) of patients proceeded to being referred to the VLCD program.
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TP 189
TP 190
COLOUR CHANGE IN CYANOSIS: RELATIONSHIP TO

CONFUSIONS OF CONGENITAL COLOUR VISION DEFICIENT
OBSERVERS
REDUCED BAROREFLEX SENSITIVITY (BRS) DURING

WAKEFULNESS IS RELATED TO SEVERITY OF SLEEP
DISORDERED BREATHING IN PATIENTS WITH OBSTRUCTIVE
SLEEP APNOEA (OSA)
R MCNAMARA1, DK MCKENZIE1, CM TAYLOR2, M CORONEO3, SJ DAIN2

Department of Respiratory and Sleep Medicine, Prince of Wales Hospital,
NSW 2031, 2School of Optometry and Vision Science, University of New
South Wales, NSW 2031, and 3Department of Ophthalmology, Prince of
Wales Hospital, NSW 2031
J NARAYAN, JR WHEATLEY, TC AMIS

Ludwig Engel Centre for Respiratory Research, Westmead Millennium
Institute; University of Sydney at Westmead Hospital, NSW, 2145
Visual recognition of cyanosis is an important clinical activity. Cyanosis recognition is affected by lighting colour and there is anecdotal evidence that people
with significant colour vision deficiencies (CVDs) have particular difficulty.
Studies to date have centred on the colour change with oxygenation of isolated
blood but it is not clear how this extrapolates to cyanotic patients in vivo.
Methods Ten patients known to be chronically hypoxaemic and showing
signs of cyanosis were recruited from the chronic respiratory program. Ten
normal subjects were recruited as controls. The spectral reflectances of their
lips, nail beds and palm creases were measured using a Topcon SR-3
telespectroradiometer. The patients were measured at rest and after exercise
to lower their saturation by 510%. The chromaticities were calculated and
plotted.
Results Both groups showed a spread of colours but they fell into two distinct
ranges. The colour difference between the groups lies very close to the colour
confusions made by congenital CVDs. Within the cyanosed group, the colour
shift was not tightly related to decreasing oxygen saturation. This is most likely
due to interpersonal factors such as pigmentation and vascular perfusion that
affect colour and the difficulties in measuring the colour of heterogeneous
anatomical features.
Conclusions These results quantify the anecdotal difficulties in detecting
cyanosis and suggest that observers with CVD would have problems recognizing the condition. The photographs obtained from this study will be used to
compare the ability of subjects with and without CVD to detect cyanosis.
Supported by the NSW Ambulance Service.
Baroreflex sensitivity is depressed in OSA patients during sleep but effects

during wakefulness are less clear. We have now examined relationships
between awake BRS and severity of sleep disordered breathing (SDB).
Methods Immediately prior to overnight polysomnography, continuous (5
min) beat-to-beat arterial blood pressure was measured via finger plethysmography (Portapres) and heart rate via ECG in 20, supine, normotensive,
untreated OSA patients (17 males; age: 49 15 years (mean SD); BMI:
26 11 kg/m2). Spontaneous baroreflex sensitivity (BRS) was calculated
using the sequence technique. SDB was characterized as apnoea hyponoea
index (events/hour) and arousal index (AI). Data were analysed via mathematical modelling and unpaired t test.
Results BRS fell with increasing AHI. Patients with AHI > 30 events/hour
(n = 9) had a significantly lower BRS (8.1 1.5 ms/mmHg) than those with
AHI < 30 events/hour (19.8 8.7 ms/mmHg, p < 0.001). BRS was negatively
related to both AHI and AI via fitted exponential functions (r2 = 0.45 and 0.70,
respectively).
Conclusion We conclude that awake BRS is lower in patients with OSA
according to the severity of SDB, particularly the number of arousals. Blood
pressure control in OSA patients during wakefulness may be impaired in
proportion to the amount of sleep disruption.
Supported by NHMRC of Australia
Nomination John Read Prize for Sleep & Physiological Research.
TP 191
APPLICATION OF MULTIVARIATE ECG ANALYSIS FOR

EVALUATION OF APNOEA HYPOPNOEA INDEX IN
POLYSOMNOGRAPHIC STUDIES
K NILSEN1,2, E ZILBERG2, D BURTON2, AH KHANDOKER1,
M PALANISWAMI1
Dept. of Electrical and Electronic Engineering, Melbourne University, VIC
3010, and 2Medical innervations, Compumedics, 30-40 Flockhart Street,
Abbotsford, Melbourne, VIC 3067
It is hypothesized that the analysis of morphology of the ECG waveform in
combination with the heart rate patterns could lead to the possibility of detection of the start and duration of apnoea/hypopnoea events and consequently
estimation of the apnoea-hypopnoea index (AHI). To the authors knowledge
the published ECG based algorithms for detecting sleep disordered breathing
are only capable of minute by minute analysis rather than detection of individual respiratory events.
Methods Changes to ECG parameters were investigated during respiratory
events with no distinction made between apnoea and hypopnoea events. 632
isolated respiratory events and 1264 controls of identical duration were
obtained from 7 polysomnographic studies, using a randomized procedure.
Features such as the R wave amplitude, T wave amplitude, QRS area and the
R-R interval were extracted from the 2 lead ECG. A number of physiological
predictors based on these features were generated. A logistic regression model
was used to investigate the association between the predictors and true
events, using the statistical software, Stata.
Results Univariate and multivariate analyses were performed. Three multivariate models were developed; heart parameters only, ECG waveform morphology parameters only and the combinations of the two. The area under the
receiver operator characteristic curves (AUC) for these models were compared. The best results were obtained with the combination of morphology and
heart rate parameters (AUC = 0.8858 (0.0078 (SD))) compared to the morphology (AUC = 0.8169 (0.0121 (SD))) and heart rate (AUC = 0.7195 (0.0103
(SD))) models.
Conclusion The multivariate analysis has shown encouraging results indicating that an algorithm using a combination of heart rate and ECG morphological parameters could potentially be constructed that would enable the
determination of individual respiratory events and subsequently an AHI.
Supported by the ARC.
TP 192
GENDER INFLUENCES ON MANDIBULAR ENCLOSURE

VOLUMES
R PERRI1,2, JR WHEATLEY1,2, TC AMIS1,2
Ludwig Engel Centre for Respiratory Research, Westmead Millennium
Institute, and 2University of Sydney at Westmead Hospital, Westmead, NSW
2145
Both male gender and increased mandibular enclosure volume predict more
severe sleep disordered breathing in obstructive sleep apnoea patients. We
now examine gender/body size/mandibular enclosure volume relationships for
normal subjects.
Methods We measured body size and cranial dimensions in 103 awake,
seated, healthy volunteers (48 males; age: 1873 yrs; BMI: 1745 kg/m2).
Calliper measured distances between skin surface cephalometric landmarks
(lateral condyle, mastoid process, gonion, gnathion, sub-nasion) were used to
reconstruct (computer software, Matlab) two three-dimensional volumes: (1)
mandibular enclosure volume (MV); and (2) retromandibular volume (RMV,
posterior to mandibular rami). Stepwise multiple linear regression analysis was
used to model body size/enclosure volume interactions.
Results For the whole group, MV was 261.1 6.0 ml (mean SE) while
RMV was 205.1 4.9 ml. Head circumference (positive) and forehead height
(negative) were both independent predictors for MV and RMV (both p < 0.02),
while hip circumference was an additional positive predictive factor for RMV
(p < 0.04). After adjusting for these parameters, male MV and RMV were larger
than for females (50.9 10.4 ml and 47.2 8.6 ml, respectively; both
p < 0.001).
Conclusion These findings suggest that mandibular enclosure volumes are
relatively larger in males, even after adjusting for body size/cranial dimension.
Differing body size/mandibular enclosure volume interactions may contribute to
gender influences on the severity of sleep disordered breathing.
Supported by NHMRC of Australia
Nomination John Read Prize for Sleep and Physiological Research.
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TP 193
VARIATION IN TIDAL VOLUMES DURING MULTI BREATH

NITROGEN WASHOUT (MBNW) AFFECTS VENTILATION
HETEROGENEITY PARAMETERS IN NORMALS AND
ASTHMATICS
TP 194
PLEURAL EFFUSION MAY CAUSE SHORTENING AND

REDUCED MOBILITY OF THE IPSILATERAL DIAPHRAGM
DJ SMITH1, K BOWDEN2, T LLOYD2, J COUCHER2, L GARSKE1
Respiratory Medicine, and 2Radiology, Princess Alexandra Hospital,
Brisbane, Australia
M SANDEMAN3, M WAINSTEIN2, R SCHOEFFEL3, N BEREND1,2,5,

C SALOME1,2,5, S DOWNIE1,2,5, B THOMPSON4,5, G KING1,2,3,5
1
The Woolcock Institute of Medical Research, 2University of Sydney 2006,
3
Department of Respiratory Medicine, Royal North Shore Hospital, St
Leonards 2065, 4Department of Allergy and Respiratory Medicine, The Alfred
Hospital, Prahran, and 5Cooperative Research Centre for Asthma and
Airways, Camperdown 2050, Australia
Introduction Sacin and Scond are measures of ventilation heterogeneity in
acinar and conducting airways, derived from analysis of MBNW. Maintaining
tidal volumes of 1 L at 911 breaths/minute (bpm) is impossible for some. Our
aim was to examine the effect of different tidal volumes on Sacin and Scond in
normals and asthmatics.
Methods 10 normals (2341 yrs) and 12 asthmatics (2163 yrs) underwent
MBNW at tidal volumes of 500 ml at 2023 bpm, 1 L at 911 bpm, and 2 L at
57 bpm. Scond and Sacin, were determined from the normalized phase III
slopes of breaths between turnovers (cumulative ventilation/FRC) 1.5 & 6.
Results The mean SD %predicted FEV1 was 97.3 17% in normals and
88 11% in asthmatics. In normals, Sacin at TV of 0.5, 1 and 2 L were
0.195 0.105 L-1, 0.095 0.036 L-1 and 0.058 0.031 L-1, respectively
(p = 0.0003, ANOVA), while Scond were 0.098 0.047 L-1, 0.042 0.021 L-1
and 0.029 0.014 L-1 (p = 0.0002), respectively. In asthmatics, Sacin were
0.440 0.195 L-1, 0.181 0.087 L-1 and 0.100 0.047 L-1, respectively
(p < 0.01), while Scond were 0.204 0.111 L-1, 0.068 0.037 L-1 and
0.031 0.013 L-1, respectively (p < 0.0001).
Conclusion Increasing tidal volume while maintaining the same minute ventilation during MBNW led to large decreases in Scond and Sacin in both
asthmatics and normals. This may be due to reduced inter-regional differences
in specific ventilation with greater TV. The log-log relationship between Sacin
and TV allows an adjustment to be made for variations in tidal volume.
Funding CRC for Asthma and Airways and NHMRC Project Grant #547346.
Introduction We have shown diaphragmatic flattening and decreased diaphragmatic excursion qualitatively assessed on ultrasound is strongly predictive of dyspnea severity and lower lung inflation in patients with pleural effusion.
We sought to quantitatively measure diaphragm length and movement and
determine how closely these are related to dyspnea severity and lung inflation.
Methods patients with unilateral pleural effusions had CT imaging of their
diaphragm during a measured inspiratory capacity manoeuvre. Maximal sagittal length was measured at TLC, and FRC. Patients had a Baseline Dyspnea
Index (BDI: 012) and respiratory function measured.
Results 4 patients with unilateral effusion (all right side; 3 malignant
mesothelioma, 1 inflammatory) had a mean (SD) BDI of 5.5 (2.89), and TLC of
74% (3.91) predicted.
Length, mean (SD)
At TLC, cm
At FRC, cm
Change, cm
Effusion right Side
Non-Effusion left side
24.4 (3.8)
26.2 (3.5)
1.8 (2.2)
25.9 (2.34)
30.3 (2.9)
4.4 (1.3)
The right diaphragm on the side of the effusion tended to be shorter than the
left at FRC (P = 0.08), and had a trend to reduced shortening with inspiration
(P = 0.08).
Conclusions the right diaphragm is known to be longer than the left in health.
The strong trend to a shorter and less mobile right diaphragm associated with
effusion suggests this is a potential mechanism for dyspnea. Further recruitment will enable correlation between BDI, TLC and diaphragm length and
mobility.
TP 195
SLEEP HEALTH IN TETRAPLEGIAA VICTORIAN POPULATION

SURVEY
S SPONG1, D RILEY1, J ROSS1, RJ PIERCE1, DJ BROWN1,2,
DJ BERLOWITZ1,2
1
Institute for Breathing and Sleep and 2Victorian Spinal Cord Service,
Austin Health, VIC 3084, Australia
Background The prevalence of obstructive sleep apnoea (OSA) and other
sleep disorders is increased in people living with tetraplegia. Despite this, no
comprehensive population survey has been conducted examining sleep disorders in tetraplegia.
Aim Determine the prevalence, nature and any relationships between sleep
disorders of those living with tetraplegia in Victoria.
Methods A questionnaire was distributed to all people in Victoria with tetraplegia containing questions regarding sleep, breathing and associated sleep
disorders.
Results Information regarding participants demographics, medical history,
state sleepiness (Karolinska Sleepiness Scale (KSS)), daily functioning (Functional Outcomes of Sleep Questionnaire (FOSQ)), quality of life (Assessment of
Quality of Life), and the likelihood of OSA (Multivariate Apnoea Prediction
Equation (MAPI)) was obtained. To date, 137 from 507 questionnaires have
been returned. The mean age of the sample is 46.9 yrs (Standard deviation = 13.6), with the average body mass index indicative of being overweight
(25.7 (5.2) kg/m2). On average, participants were neither sleepy nor alert
(KSS = 4.1 (2.3) and had functioning attributable to sleepiness (Total
FOSQ = 17.7 (2.4)) that was slightly worse than an able bodied, control population (17.9 (3.1)), but better than an able-bodied population with untreated
OSA (14.5 (3.6)). The MAPI predicted that 14% of the sample were likely to
have OSA. These data will be complimented by full sleep studies to be
performed at the participants homes in late 2007, early 2008.
Conclusion Our interim data suggest that the rate of subjective sleep
complaints are not substantially different in the population with tetraplegia
compared with the able-bodied.
This research was supported by the Victorian Neurotrauma Initiative.
TP 196
CONDUCTIVE VENTILATION HETEROGENEITY DOES NOT

AFFECT DIFFUSING CAPACITY MEASUREMENT
B THOMPSON2, D SCHUERMANS1, S VANMALDEREN1, W VINCKEN1,
S VERBANCK1
1
Respiratory Division, University Hospital UZ Brussel, Vrije Universiteit
Brussel, 1090 Brussels, Belgium, and 2Department of Allergy, Immunology
and Respiratory Medicine. The Alfred Hospital, Melbourne, Australia 3004
It has long been assumed that the ventilation heterogeneity associated with
lung disease could in itself affect the measurement of carbon monoxide transfer factor. The aim of this study was to investigate the potential estimation
errors of carbon monoxide diffusing capacity (TLco) measurement that are
specifically due to conductive ventilation heterogeneity. We induced conductive
airway ventilation heterogeneity in 35 never-smoker normal subjects by histamine provocation, and related the resulting changes in ventilation heterogeneity (derived from the multiple breath washout test) to corresponding changes in
diffusing capacity, alveolar volume and inspired vital capacity (derived from the
single breath TLco method). Average conductive ventilation heterogeneity
doubled (p < 0.001), while TLco decreased by 6% (p < 0.001), with no correlation between individual data (p > 0.1). When dividing diffusing capacity by
alveolar volume, the resulting transfer coefficient was not significantly different
pre versus post histamine (p = 0.074). These findings can be brought in agreement with recent modelling work, where specific ventilation heterogeneity
resulting from different distributions of either inspired volume or end-expiratory
lung volume have been shown to affect TLco estimation errors in opposite
ways. The combination of these errors appears to largely cancel out in our
experimental situation of induced ventilation heterogeneity comparable to that
observed in lung disease. We conclude that conductive ventilation heterogeneity per se has a negligible effect on diffusing capacity measurement.
A74
TP 197
ABSENCE OF AIRWAY TONE IN BALB/c MICE

D TURNER1, A LARCOMBE1, G ZOSKY1, E BOZANICH1, Z HANTOS1,2,
P SLY1
1
Telethon Institute for Child Health Research, and Centre for Child Health
Research, The University of Western Australia, Perth 6008, and 2Department
of Medical Informatics and Engineering, University of Szeged, Hungary
An important determinant of airway function in humans is vagal-mediated
cholinergic tone in airway smooth muscle (ASM). This airway tone may be
altered in disease states. The use of mouse models for the study of airway
diseases, including asthma, pulmonary fibrosis and COPD is well established.
However, it is not known whether mice actually possess basal ASM tone or, if
it does exist, how this tone changes in disease models. This study was
undertaken to determine whether mice have detectable ASM tone in vivo.
Methods Respiratory system impedance (Zrs) was measured in female adult
BALB/c mice using a wave-tube modification of the forced oscillation technique. Zrs was measured during slow (~35 s) inflation-deflation manoeuvres
between the transrespiratory pressures of 0 and 20 cmH2O. Baseline lung
mechanics and thoracic lung volumes (TGV) were measured before and after
each mouse was allocated to one of four treatment groups: saline mice
received an i.p injection of saline, atropine mice received i.p. atropine sulphate, vagotomy mice had their left and right cervical vagus nerves isolated by
blunt dissection and cut, and sham mice had the area of the vagus nerves
exposed but the nerves were not cut.
Results There were no post-treatment changes in TGV, airway resistance,
tissue damping, tissue elastance, inertance or tissue hysteresivity in any of the
four groups.
Conclusions The lack of change in lung mechanics post-atropine or postvagotomy in BALB/c mice suggests that, unlike humans and many other
species, the airways of mice have no baseline ASM tone.
Supported by NHMRC grant#11488.
Nomination None.

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Respirology (2008) 13, (Suppl.

TSANZ Poster Sessions

EARLY LIFE PNEUMOVIRAL INFECTION INDUCES THE

ANTIGEN-SPECIFIC TH1 CELLS INDUCE AIRWAY

NICOLE HANSBRO1, SARAH BOUSTANY2, BRIAN OLIVER2, JANETTE

M YANG, C MA, PS FOSTER

ABSENCE OF SPHINGOSINE KINASE 1 LEAD TO

Journal compilation 2008 Asian Pacific Society of Respirology

STREPTOCOCCUS PNEUMONIAE VACCINE PREVENAR:

Respirology (2008) 13, (Suppl. 2)

A SHEEP ASTHMA MODEL FOR COMPARATIVE STUDIES ON

The underlying biological processes involved in airway remodelling are still

RELAXIN TREATMENT REVERSES AIRWAY REMODELLING

Airway remodelling is a set of structural changes in asthma that has been

A MOUSE MODEL OF SUBLINGUAL ALLERGEN

MEASURING SEVERE LUNG DISEASE IN MICE

CHARLES L HARDY, JUN YAO, JENNIFER M ROLLAND, ROBYN E OHEHIR

ALEXANDER N LARCOMBE1, GRAEME R ZOSKY1, DEBRA J TURNER1,

Background The flexiVent small animal ventilator (SCIREQ, Canada) allows

Journal compilation 2008 Asian Pacific Society of Respirology

NEUTROPHIL INFLUX DURING CHLAMYDIAL LUNG INFECTION

HAEMOPHILUS INFLUENZAE (Hi) INFECTION IN ASTHMA MAY

JC HORVAT, KW BEAGLEY, PG GIBSON, PS FOSTER, PM HANSBRO

AMA-TAWIAH ESSILFIE, JULIE PRESTON, JAY HORVAT, MARGARET

NA is characterized by the presence of neutrophilic lung inflammation. Hi

NOVEL BRONCHODILATOR RESPONSE TO ROSIGLITAZONE

PERSISTENCE OF AIRWAY HYPERRESPONSIVENESS IN MICE

JANE WARD1, YAN BAI2, MICHAEL SANDERSON2

The ligand-activated transcription factor proliferator activated receptor gamma

Journal compilation 2008 Asian Pacific Society of Respirology

Respirology (2008) 13, (Suppl. 2)

THE RELATIONSHIP BETWEEN AIRWAY SIZE AND

VASCULAR REMODELLING IN A SHEEP MODEL OF CHRONIC

JESSICA KERMODE1,2, BRIAN OLIVER1,2,3, BRENT MCPARLAND1,2,3

Asthmatic airways are highly sensitive to a number of stimuli. The reason

It is now recognized that long-term structural and functional changes to lung

J VAN DER VELDEN1, SJ HIRST2, KJ SNIBSON1

MAST CELL DENSITIES IN ENDOBRONCHIAL BIOPSIES AND IN

Journal compilation 2008 Asian Pacific Society of Respirology

MAST CELL DENSITIES IN ENDOBRONCHIAL BIOPSIES AND IN

AGEING ALTERS AIRWAY AND CIRCULATING NEUTROPHIL

INNATE IMMUNE RESPONSES OF AIRWAY NEUTROPHILS ARE

Age is an important factor in the development of neutrophilic airway diseases

Neutrophils play a crucial role in innate immunity and contribute significantly to

AIRWAY SMOOTH MUSCLE CELL VOLUME IS UNCHANGED IN

Conclusion These data suggest that although ASM area is increased in

HUMAN LUNG MAST CELL PRODUCTS MODULATE AIRWAY

Journal compilation 2008 Asian Pacific Society of Respirology

Respirology (2008) 13, (Suppl. 2)

TNF-a AND IFN-g DIFFERENTIALLY REGULATE AIRWAY

A MODEL OF IgE SENSITIZATION TO STUDY RHINOVIRUS

DAVID I KRIMMER1, MATAKITAU LOSELI1, J MARGARET HUGHES2,

M LINDSAY1,2, B THOMAS1,2, H DAGHER1,2, J MEANGER1,2, R GHILDYAL2,

Chronic airway inflammation in asthma is maintained by provision of survival

The technique of IgE passive sensitization reproduces IgE-related allergic

MMP AND TIMP PRODUCTION BY CULTURED AIRWAY SMOOTH

CIRCULATING EOSINOPHIL COUNTS ARE HIGHER AFTER A

PETER BLACK, SUSANNE BRODIE, CLAIRE ARANDJUS, FAY SOMMERVILLE,

Journal compilation 2008 Asian Pacific Society of Respirology

IMPAIRMENT OF AIRWAY MACROPHAGE PHAGOCYTOSIS OF

INTRAVENOUS IgG AMELIORATES ALLERGIC AIRWAY

CHRISTINE VAN DALEN1, ELIZABETH HARDING1, PRACHEE GOKHALE1,

M YAMAMOTO1, K KOBAYASHI1, Y ISHIKAWA1, K NAKATA1, E