Development of wheat lines carrying stem rust resistance gene Sr39 with reduced Aegilops speltoides chromatin and simple PCR markers for marker-assisted selection.

The use of major resistance genes is a cost-effective strategy for preventing stem rust epidemics in wheat crops. The stem rust resistance gene Sr39 provides resistance to all currently known pathotypes of Puccinia graminis f. sp. tritici (Pgt) including Ug99 (TTKSK) and was introgressed together with leaf rust resistance gene Lr35 conferring adult plant resistance to P. triticina (Pt), into wheat from Aegilops speltoides. It has not been used extensively in wheat breeding because of the presumed but as yet undocumented negative agronomic effects associated with Ae. speltoides chromatin. This investigation reports the production of a set of recombinants with shortened Ae. speltoides segments through induction of homoeologous recombination between the wheat and the Ae. speltoides chromosome. Simple PCR-based DNA markers were developed for resistant and susceptible genotypes (Sr39#22r and Sr39#50s) and validated across a set of recombinant lines and wheat cultivars. These markers will facilitate the pyramiding of ameliorated sources of Sr39 with other stem rust resistance genes that are effective against the Pgt pathotype TTKSK and its variants.

Which obese children should have a sleep study?

BACKGROUND: The UK government has recommended the development of obesity services for children. As obesity is common, studying every obese child for obstructive sleep apnoea (OSA) would be challenging and full paediatric sleep services are not available in every area in Europe. The purpose of this study was to consider how well clinical features predict significant OSA in obese children in order to help prioritise the need for sleep studies and subsequent treatment. METHODS: Consecutive children referred for obesity management aged 2-16 years with a body mass index (BMI) of >2.5 z scores for age were offered a sleep study using overnight oximetry and audiovisual recordings. Significant OSA was defined as > or = 5 dips/h of >4% oxygen saturation or > or = 5 respiratory-event related arousals/h. RESULTS: Forty-one of 158 (26%) children (mean BMI z score 3.7) had significant OSA and 95% of these had either reported apnoea, restless sleep or tonsillar hypertrophy (TH). Nineteen percent of all children had none of these features. BMI was not related to OSA. CONCLUSION: If only obese children with reported apnoea, restless sleep or TH have a sleep study, 95% of all obese children with significant OSA will be identified using this method.

Construction of comparative genetic maps of two 4Bs.4Bl-5Rl translocations in bread wheat (Triticum aestivum L.).

The physical length of the rye segment of a 4BS.4BL-5RL translocation derived from the Cornell Wheat Selection 82a1-2-4-7 in a Triticum aestivum 'Chinese Spring' background was measured using genomic in situ hybridization (GISH) and found to be 16% of the long arm. The size of this translocation was similar to previously published GISH measurements of another 4BS.4BL-5RL translocation in a Triticum aestivum 'Viking' wheat background. Molecular maps of both 4BS.4BL-5RL translocations for 2 different wheat backgrounds were developed using RFLP analysis. The locations of the translocation breakpoints of the 2 4BS.4BL-5RL translocations were similar even though they arose in different populations. This suggests a unique property of the region at or near the translocation breakpoint that could be associated with their similarity and spontaneous formation. These segments of rye chromosome 5 also contain a gene for copper efficiency that improves the wheat's ability to cope with low-copper soils. Genetic markers in these maps can also be used to screen for copper efficiency in bread wheat lines derived from the Cornell Wheat Selection 82a1 2-4-7.

Single nucleotide polymorphic marker enabling rapid and early screening for the homoeolocus of beta-amylase-R1: a gene linked to copper efficiency on 5RL.

This study describes the development of a PCR marker to detect the beta-amylase-R1 gene of rye. It provides an easy and rapid means for the identification of plants containing the beta-amylase-R1. Because rye chromosome segments do not normally recombine with wheat chromosomes, this marker provides a means for tracking all linked genes on that alien 5RL chromosome segment. Reaction conditions were optimised for an annealing temperature of 60 degrees C for a high stringency. The reaction was also optimised for low reaction volumes reducing the cost of the reagents required for the reaction. This PCR test can be used in breeding or mapping programs for the rapid screening of progeny containing translocations of 5RL and hence select for the copper efficiency trait of rye.

Development of PCR markers for the selection of wheat stem rust resistance genes Sr24 and Sr26 in diverse wheat germplasm.

The use of major resistance genes is the most cost-effective strategy for preventing stem rust epidemics in Australian wheat crops. The long-term success of this strategy is dependent on combining resistance genes that are effective against all predominant races of the pathogen, a task greatly assisted by the use of molecular markers linked to individual resistance genes. The wheat stem rust resistance genes Sr24 and Sr26 (derived from Agropyron elongatum) and SrR and Sr31 (derived from rye) are available in wheat as segments of alien chromosome translocated to wheat chromosomes. Each of these genes provides resistance to all races of wheat stem rust currently found in Australia . We have developed robust PCR markers for Sr24 and Sr26 (this study) and SrR and Sr31 (previously reported) that are applicable across a wide selection of Australian wheat germplasm. Wheat lines have recently become available in which the size of the alien segments containing Sr26, SrR and Sr31 has been reduced. Newly developed PCR-markers can be used to identify the presence of the shorter alien segment in all cases. Assuming that these genes have different gene-for-gene specificities and that the wheat industry will discourage the use of varieties carrying single genes only, the newly developed PCR markers will facilitate the incorporation of two or more of the genes Sr24, Sr26, SrR and Sr31 into wheat lines and have the potential to provide durable control to stem rust in Australia and elsewhere.

Skin-prick testing as a diagnostic aid for childhood asthma.

Diagnosing asthma is problematic when based solely on reported symptoms. The purpose of this study was to evaluate skin-prick testing as a diagnostic aid for asthma in children. Skin-prick testing (SPT) was undertaken in children aged 2-10 years with either no history of wheeze (n = 149) or recent doctor-observed wheeze which responded to treatment with a bronchodilator, the "gold standard" (n = 164). Children with moderate or severe asthma were excluded. SPT positivity increased sharply at age 5 years in wheezers. Data were therefore divided into two age groups: 2- < 5 years (57 controls, 97 wheezers) and 5-10 years (92 controls, 67 wheezers). The sensitivity, specificity, and likelihood ratios of SPT positivity for wheeze were 32%, 89%, and 2.9, respectively, in the younger children, and 82%, 85%, and 5.5, respectively, in the older children. For a prevalence of 30% for asthma, the positive predictive values of a positive SPT were 55% and 70% for the younger and older age groups, respectively. The test characteristics of SPT for helping diagnose asthma in schoolchildren are good. The prevalence of wheeze in preschool children is high, and so SPT should be helpful even in this group. We suggest that clinicians consider skin-prick testing as a diagnostic aid for asthma.

Diagnostic accuracy of bronchodilator responsiveness in wheezy children.

BACKGROUND: The aim of this study was to evaluate the diagnostic accuracy of bronchodilator responsiveness for asthma in 5-10 year old children. METHODS: Spirometric measurements were made in 142 children (58 wheezers) before and after 400 mug inhaled salbutamol. RESULTS: On a receiver operating characteristic curve, a 9% increase in predicted forced expiratory volume in 1 second was the cut off point that provided an acceptable balance of sensitivity and specificity for previous wheeze. This figure was 50% (95% CI 38 to 62) sensitive and 86% (95% CI 78 to 92) specific for detecting previous wheeze and multiplied the initial odds in favour of wheeze by a factor of 3.6 (95% CI 2.0 to 6.3). DISCUSSION: With an estimated pretest probability of wheeze of 10% in the community and 50% in a specialist clinic, the positive predictive values are 29% and 78%, respectively, for a 9% change. The value of bronchodilator responsiveness testing depends on the prevalence of wheeze in the population in which it is to be used.

Repeatability of airway resistance measurements made using the interrupter technique.

BACKGROUND: To be able to interpret any measurement, its repeatability should be known. This study reports the repeatability of airway resistance measurements using the interrupter technique (Rint) in children with and without respiratory symptoms. METHODS: Children aged 2-10 years who were healthy, had persistent isolated cough, or who had previous wheeze were studied. On the same occasion, three Rint measurements were made 15 minutes apart, before and after placebo and salbutamol given in random order. Results from those given placebo first were analysed for within-occasion repeatability. Between-occasion repeatability measurements were made 2-20 weeks apart (median 3 weeks). RESULTS: For 85 pairs of measurements before and after placebo the limits of agreement were 20% expected resistance and were unaffected by age or health status. The change in resistance following bronchodilator in one of 18 healthy children, 12 of 28 with cough, and 22 of 39 with wheeze exceeded this threshold. For between-occasion measurements the limits of agreement were 32% in 72 healthy subjects, 49% in 57 with cough, and 53% in 95 with previous wheeze. CONCLUSION: The measurement of airways resistance by the interrupter technique is clinically meaningful when change following an intervention such as the administration of bronchodilator is greater than its within-occasion repeatability. Between-occasion repeatability is too poor to judge change confidently.

Airway resistance measured by the interrupter technique: normative data for 2-10 year olds of three ethnicities.

BACKGROUND AND AIMS: The measurement of airway resistance using the interrupter technique (R(int)) is feasible in preschool children and other subjects unable to undertake spirometry. This makes it potentially useful for the measurement of lung function in these groups. Commercial devices use different algorithms to measure pressure and flow from which R(int) is derived. This study provides normative values for British children using devices from a single manufacturer. METHODS: R(int) was measured in 236 healthy children of three ethnic groups (Afro-Caribbean and black African, Bangladeshi, and white British) aged 2-10 years using Micro Medical devices. Software in the devices calculated R(int) from pressure measured by the two point, back extrapolation method from the pressure transient during valve closure, with flow measured just before valve closure. RESULTS: R(int) is related to both age and height, but when age is allowed for there is not a significant relation with height. Neither gender nor any of the ethnicities studied was significantly related to R(int). DISCUSSION: These measurements in healthy children using this technique may be used as reference data for similar populations.

Airway function at one year: association with premorbid airway function, wheezing, and maternal smoking.

BACKGROUND: Impaired growth and development of the respiratory system during fetal and early postnatal life may have important implications for lung development and later lung health. The aim of this study was to examine the association of diminished premorbid airway function, prior wheezing, and maternal smoking with airway function at 1 year of age. METHODS: Respiratory function was measured at the end of the first year in 100 of 108 healthy term infants (93%) in whom similar measurements had been undertaken prior to any respiratory illness at 8 weeks. Physician diagnosed wheezing episodes were identified retrospectively from medical records. RESULTS: At 1 year specific airway conductance during end expiration (sGawEE; /s/kPa) was significantly diminished in those infants with prior wheezing (95% CI wheeze/no wheeze -0.76 to -0.14), mothers who smoked (95% CI smoke/no smoke -0.81 to -0.27), a family history of asthma (95% CI family history/no family history -0.62 to 0.00), or diminished premorbid sGawEE (95% CI -0.13 to -0.43/s/kPa per unit reduction sGawEE at 8 weeks). In a multivariate model only maternal smoking and diminished premorbid sGawEE were independently associated with diminished sGawEE at 1 year. CONCLUSIONS: Diminished airway function at the end of the first year appears to be mediated by impaired airway development during early life as well as by exposure to maternal smoking. These findings are consistent with the hypothesis that, at a population level, diminished premorbid airway function provides the link between wheezing lower respiratory illness and diminished airway function at 1 year. Maternal smoking remains an important and avoidable cause of impaired airway development and function in infancy.

Respiratory function in survivors of the United Kingdom Extracorporeal Membrane Oxygenation Trial.

Extracorporeal membrane oxygenation (ECMO) improves survival in mature neonates with reversible lung disease. However, ECMO could result in survival of infants with severe respiratory dysfunction who would otherwise have died. Alternatively, infants receiving ECMO might be spared prolonged ventilation and consequent barotrauma, resulting in improved respiratory function. Our aim was to compare respiratory function at 1 yr of age in infants assigned to receive either ECMO or conventional management (CM). Seventy-eight surviving infants of the United Kingdom (UK) ECMO trial (51 in the ECMO group) were studied at 1 yr of age. Questionnaires provided details of respiratory symptoms, and laboratory measurements of respiratory function were made for respiratory rate, tidal volume, lung volume, airway conductance, specific airway conductance, and maximal expiratory flow at FRC (Vmax (FRC)). Data were exchanged on floppy disk for cross-analysis and to ensure that investigators were blinded to the status of the infants. There was a wide spectrum of respiratory function, from normal to markedly abnormal. There were few differences between the groups, but in the CM group lung volume was increased (95% confidence intervals [CIs] of the difference in ECMO versus CM subjects: -67; -4 ml), and inspiratory specific conductance was lower (95% CI: 0.03; 0.98 s(-)(1). kPa(-)(1)). There was a trend toward a lower V max(FRC) (95% CI: -2; 67 ml/s(-)(1) in the CM group. In addition to providing a survival advantage, ECMO did not worsen lung function in infants assigned to receive it. Indeed, their lung function appeared slightly better than that of infants treated conventionally.

Impaired airway function and wheezing in infancy: the influence of maternal smoking and a genetic predisposition to asthma.

This prospective community-based study of infants born in inner London was undertaken to examine the association between premorbid airway function and subsequent wheezing in the first year of life and to explore the influence on this association of a family history of asthma and maternal smoking during pregnancy. Healthy Caucasian term infants were recruited shortly after birth, and physician-diagnosed wheezing episodes were identified retrospectively from medical records. Specific airway conductance was determined from plethysmographic measurements of lung volume and airway resistance, before 13 wk and prior to any respiratory illness, in 101 infants, 28 of whom experienced at least one episode of wheezing during the first year. Mean (SD) specific airway conductance was significantly diminished in infants who subsequently wheezed: 2.02 (1.07) s-1. kPa-1 and 2.60 (0.93) s-1. kPa-1, respectively (p < 0.05), and in those with a first-degree relative with asthma: 1.98 (0.83) s-1. kPa-1 and 2.60 (1.0) s-1. kPa-1, respectively (p < 0.05), but not in those whose mothers smoked during pregnancy, in whom airway resistance was, however, significantly elevated (p < 0.05). The odds ratio (95% confidence interval [CI]) for wheezing was 2.1 (1.1 to 3.8) for every unit (s-1. kPa-1) decline in specific airway conductance (p = 0.02). After adjustment for premorbid airway function, the odds of wheezing were significantly increased in those with a family history of asthma (4. 3; 95% CI, 1.3 to 13.8; p = 0.016) and those exposed to maternal smoking during pregnancy (4.9; 95% CI, 1.6 to 15.0; p = 0.005). Our findings confirm previous reports that impaired premorbid airway function precedes and predicts wheezing in the first year. Among those with a genetic predisposition to asthma, alterations in airway geometry or tone may increase susceptibility to wheezing. Maternal smoking has important and potentially preventable adverse effects on somatic growth and respiratory morbidity in early life.

Distinct patterns of respiratory difficulty in young children with achondroplasia: a clinical, sleep, and lung function study.

AIM: Achondroplasia can result in respiratory difficulty in early infancy. The aim of this study was to document lung growth during infancy, together with the cause of any cardiorespiratory and sleep dysfunction. PATIENTS AND METHODS: Seventeen prospectively ascertained infants (14 boys and three girls) with respiratory symptoms starting before 1 year of age underwent clinical, sleep, and lung function studies. RESULTS: Three distinct groups were identified. Group 1 (n = 6) were the least symptomatic and only had obstructive sleep apnoea. Group 2 (n = 6) had obstructive sleep apnoea of muscular aetiology and, neurologically, hydrocephalus and a small foramen magnum were common. Group 3 (n = 5), the most severely affected group, all developed cor pulmonale, with three deaths occurring as a result of terminal cardiorespiratory failure. All five had obstructive sleep apnoea with a muscular aetiology (a small foramen magnum predominated) with severe or moderately severe gastro-oesophageal reflux. Initially, lung function studies found no evidence of restriction or reduced lung volumes standardised according to weight. However, with growth these infants had worsening function, with raised airway resistance and severe reductions in respiratory compliance. CONCLUSIONS: These groups appear to be distinct phenotypes with distinct anatomical aetiologies: "relative" adenotonsillar hypertrophy, resulting from a degree of midfacial hypoplasia (group 1); muscular upper airway obstruction along with progressive hydrocephalus, resulting from jugular foramen stenosis (group 2); and muscular upper airway obstruction, but without hydrocephalus, resulting from hypoglossal canal stenosis with or without foramen magnum compression and no jugular foramen stenosis (group 3). The aetiology of these abnormalities is consistent with localised alteration of chondrocranial development: rostral, intermediary and caudal in groups 1, 2, and 3, respectively.

A collaborative study of infant respiratory function testing.

The aims of this study were to compare inter-observer variability within and between two specialized infant lung function testing centres and to develop a strategy for performing and analysing infant respiratory function tests to facilitate future collaborative trials. A protocol for data collection and analysis was developed using similar equipment and identical software. All raw data were exchanged on disk and analysed, blind to infant status. All data were cross-analysed by both centres to assess inter-observer variability. Outcome measures were functional residual capacity (FRCpleth), airway resistance (Raw) and maximal expiratory flow at FRC (V'max,FRC). Subjects were recruited from the multicentre UK extracorporeal membrane oxygenation (ECMO) Trial and measured at around 1 yr of age. Forty-two infants attended the Institute of Child Health, London and 36 attended the Leicester Royal Infirmary. The proportion of infants treated with ECMO or conventional management at each centre was similar. There were no significant differences between any of the outcome measures for infants tested at either centre. During a cross-analysis, the agreement between the two centres, within infant, was closer for V'max,FRC and FRCpleth (within 10%) than for the more variable measurements of Raw (within 20%). A collaborative approach to trials with infant respiratory function as an outcome measure appears feasible, providing that close attention is paid to study design, and participants in such trials maintain a standard approach to data collection and analysis.

Infant respiratory function after RSV-proven bronchiolitis.

The mechanisms underlying the increased risk of wheezing in early childhood following acute bronchiolitis in infancy remain unclear. Previous studies have reported significant abnormalities in infant respiratory function after clinical recovery from bronchiolitis, but are difficult to interpret because of the frequent omission of a concurrent comparison group. Respiratory function was compared within pairs of previously healthy full-term caucasian infants admitted with a first episode of acute bronchiolitis to an inner London hospital, and age- and sex-matched control infants without prior wheezing, asthma, or lower respiratory illness who were recruited from local general practices. Respiratory function was measured in 29 control and 29 asymptomatic index infants, with measurements in the latter done at a median interval of 36 wk (range: 16 to 49 wk) after admission, when 16 (55%) had experienced subsequent wheezing. Index infants tended to be autumn-born and of shorter gestation than control infants, to have younger mothers, and to have been exposed to tobacco smoke. There were no statistically significant differences in plethysmographic FRC, initial inspiratory airway resistance (Raw), or respiratory system compliance (mean [index minus control] within-pair difference [95% confidence interval]: -11 ml [-29, 7 ml]; -0.2 kPa/L/s [-0.7, 0.4 kPa/L/s]; -8 ml/kPa [-21, 4 ml/kPa], respectively), but respiratory rate and time to peak tidal flow as a proportion of total expiratory time (tPTEF:tE) were significantly diminished in index as compared with control infants (-4.0 breaths/min [-7.6, -0.4 breaths/min], versus -0.035 [-0.066, -0.005], respectively). These findings suggest a better prognosis for infant lung function after acute bronchiolitis than reported previously. Longitudinal studies are needed to clarify whether subclinical alterations in airway function precede acute bronchiolitis.

A critical assessment of uncalibrated respiratory inductance plethysmography (Respitrace) for the measurement of tidal breathing parameters in newborns and infants.

We have compared results obtained with an uncalibrated respiratory inductance plethysmograph (RIP) with those of a face mask and pneumotachograph (PNT) for the computerized measurement of the time to reach peak tidal expiratory flow as a ratio of total expiratory time (tPTEF:tE). Simultaneous measurements were made in 32 healthy neonates aged 0-3 weeks, 35 healthy infants aged 5-82 weeks, and 28 infants aged 15-94 weeks with physician diagnosed recurrent wheeze. The group mean (+/- SD) values of tPTEF:TE determined using a PNT were 0.455 (+/- 0.129), 0.263 (+/- 0.077), and 0.232 (+/- 0.089) for the neonates, healthy infants and infants with recurrent wheeze respectively. RIP gave mean (+/- SD) values that were 0.055 (+/- 0.044) and 0.025 (+/- 0.104) lower than the PNT in healthy neonates and infants with recurrent wheeze respectively; RIP values were 0.002 (+/- 0.073) higher in the healthy infants over 4 weeks of age than measurements by PNT. Although the difference between the two measurements was not related to the thoracoabdominal phase angle, as measured from Lissajous figures, examination of the RIP ribcage and abdominal signals revealed that many healthy subjects, while appearing clinically in phase, had ribcage and abdominal signals that differed markedly from each other in terms of convexity/concavity during early expiration. This may explain the lack of agreement between the two methods. We conclude that uncalibrated RIP should be used with caution for the determination of tPTEF:tE, even in subjects whose ribcage and abdomen appear to move synchronously. The measurement of tPTEF:tE did not differentiate between the healthy infants and infants with recurrent wheezing.

The single breath test in neonates: does pressurization of the pneumotachograph make a difference?

The single breath test for the measurement of respiratory system resistance and compliance in newborns consists of an end inspiratory occlusion which is subsequently released, allowing expiration to proceed through a pneumotachograph (PNT). The measured flow is then integrated to give volume. The simplicity of the test is one of the major reasons for its popularity. However, some investigators have cautioned against the use of an occlusion distal to the PNT because pressurization of the PNT may introduce artifacts in the flow measurement. Despite this caution, many commercial systems use a pressurized PNT. This study investigated the errors that would result from pressurization of the PNT by providing a step function of flow to two infant PNTs, a Fleisch #0 and a Hans Rudolph 4500, in the unpressurized and pressurized state. In each case there was an initial rapid rise of the flow signal, followed by some overshoot and oscillations that rapidly died out. The overshoot and oscillations for the Hans Rudolph PNT were greater when pressurized whereas pressurization had little effect on the Fleisch PNT. Unpressurized, the two were similar. In either case, the artifact introduced by pressurization of the PNT died out so quickly that it would have little effect on any measurement in an infant.