Printed three-dimensional airway model assists planning of single-lung ventilation in a small child.

BACKGROUND: Single-lung ventilation in infants and small children is challenging because suitable sizes of double-lumen cuffed tracheal tubes are not available. A 6-yr-old child required pulmonary saline washout for primary alveolar proteinosis, and therefore needed sequential single-lung ventilation in order to achieve safe oxygenation. Before undertaking this potentially hazardous procedure, we practised bronchial intubation on an anatomical model of her airway constructed from computed tomography (CT) data. METHODS: We created a full-scale, anatomically accurate, transparent plastic model of the trachea and main bronchi on a three-dimensional printer using data from a CT scan. We then performed several different airway approaches to identify those likely to be most suitable, ex vivo, before the clinical procedure was carried out on the patient. RESULTS: The model helped us to choose the type and size of bronchial tubes and to practise their insertion beforehand. Subsequently, during anaesthesia, the chosen technique was successful. CONCLUSIONS: Three-dimensional printing of a model of the airway of a small child aided planning of bronchial intubation and single-lung ventilation. Three-dimensional printing of airway structures may have wider application in anaesthesia practice.

The use of cuffed tracheal tubes for paediatric tracheal intubation, a survey of specialist practice in the United Kingdom.

BACKGROUND AND OBJECTIVE: For more than 50 yr, uncuffed tracheal tubes have been the gold standard for intubation in children under the age of 8 yr. However, recently there has been interest in the use of cuffed tubes in paediatric practice. This survey aimed to benchmark UK practice with regard to tracheal intubation within specialist paediatric centres, exploring current cuffed tracheal tube use in children. METHODS: A questionnaire was e-mailed to the paediatric intensive care unit and anaesthetic department clinical leads in all UK specialist paediatric hospitals with a paediatric intensive care unit (n = 30). Information was requested on the use of tracheal tubes across all paediatric age groups, as well as the reasons for non-use and the incidence of complications attributed to cuffed tubes. RESULTS: A total of 20 paediatric intensive care unit and 15 anaesthetic questionnaires were returned, equating to a response rate of 67% and 50%, respectively. Only 5% of the paediatric intensive care unit and 7% of the anaesthetic respondents routinely use a cuffed tube in children under the age of 8 yr. The commonest reason cited in both groups for non-cuff use was that there is minimal benefit to be gained over using an uncuffed tracheal tube. The most frequent specific indication for use of a cuffed tube was a reduced lung compliance (60% respondents both groups). In all, 45% of the paediatric intensive care unit respondents and 100% of the anaesthetists reported that they did not routinely monitor the intracuff pressure when using a cuffed tube. The incidence of observed complications attributed to the use of cuffed tubes was far higher amongst paediatric intensive care unit consultants (65% vs. 7% anaesthetists); however, the majority in both groups stated that such complications were no more common than when using an uncuffed tube (60% paediatric intensive care unit and 53% anaesthetists). CONCLUSION: Cuffed tracheal tubes are rarely routinely used in children, particularly in the under 8 yr age group, in specialist paediatric centres in the UK. When used, it is predominantly for a specific indication, and the monitoring of intracuff pressure is not routine. Current expert consensus is that complications are equally as common when using a cuffed as an uncuffed tube.

An evaluation of the sensitivity and specificity of energy expenditure measured by heart rate and the Goldberg cut-off for energy intake: basal metabolic rate for identifying mis-reporting of energy intake by adults and children: a retrospective analysis.

OBJECTIVE: To identify adults and children as under- (UR), acceptable (AR), or over-reporters (OR) of energy intake (EI) using energy expenditure measured by doubly labelled water (DLW) (EE(DLW)), and to use this as a reference to determine the sensitivity and specificity of (i) EE measured by heart rate (EE(HR)), and (ii) the Goldberg cut-off technique for classifying subjects into the same categories. DESIGN: Retrospective analysis of a dataset comprising concurrent measurements of EE(DLW), EE(HR), basal metabolic rate (BMR), and EI by weighed record (EI(WR)) on 14 adults and 36 children. EI by diet history (EI(DH)) was also measured in the children only. EI(WR):EE(DLW) provided the reference definition of subjects as UR, AR or OR. Three strategies for classifying mis-reporters based on EE(HR) and Goldberg cut-offs were then explored. Sensitivity and specificity were calculated respectively as the proportion of UR and non-UR correctly identified. RESULTS: Approximately 80% of all subjects were AR. For EI(WR) and EI(DH) respectively, the sensitivity of EE(HR) was 0.50 and 1.00, and specificity was 0.98 and 1.00. Although designating subjects as having low, medium or high activity levels (EE(HR):BMR(meas)) and calculating cut-offs based on appropriate WHO physical activity level PALs did not change sensitivity, specificity dropped to 0.98 (EI(WR)) and 0.97 (EI(DH)). Cut-offs based on a PAL of 1.55 reduced sensitivity to 0.33 (EI(WR)) and 0.00 (EI(DH)), but specificity remained unchanged. The sensitivity of all cut-offs based on physical activity level (PALs) for EI(WR) was 0.50 (adults) and 0.25 (children). CONCLUSIONS: If the precision of EE(HR) was improved, it may be useful for identifying mis-reporters of EI.

The genetic basis of plasma variation in adiponectin, a global endophenotype for obesity and the metabolic syndrome.

Here we present the first genetic analysis of adiponectin levels, a newly identified adipocyte-derived protein. Recent work has suggested that adiponectin may play a role in mediating the effects of body weight as a risk factor for coronary artery disease. For this analysis we assayed serum levels of adiponectin in 1100 adults of predominantly northern European ancestry distributed across 170 families. Quantitative genetic analysis of adiponectin levels detected an additive genetic heritability of 46%. The maximum LOD score detected in a genome wide scan for adiponectin levels was 4.06 (P = 7.7 x 10(-6)), 35 cM from pter on chromosome 5. The second largest LOD score (LOD = 3.2; P = 6.2 x 10(-5)) was detected on chromosome 14, 29 cM from pter. The detection of a significant linkage with a quantitative trait locus on chromosome 5 provides strong evidence for a replication of a previously reported quantitative trait locus for obesity-related phenotypes. In addition, several secondary signals offer potential evidence of replications for additional previously reported obesity-related quantitative trait loci on chromosomes 2 and 10. Not only do these results identify quantitative trait loci with significant effects on a newly described, and potentially very important, adipocyte-derived protein, they also reveal the emergence of a consistent pattern of linkage results for obesity-related traits across a number of human populations.

Radiation hybrid mapping of 11 alpha and beta nicotinic acetylcholine receptor genes in Rattus norvegicus.

Acetylcholine is the main neurotransmitter of the vestibular efferents and a wide variety of muscarinic and nicotinic acetylcholine receptors are expressed in the vestibular periphery. To date, 11 nicotinic subunits (alpha and beta) have been reported in mammals. Previously, our group [Brain Res. 778 (1997) 409] reported that these nicotinic acetylcholine receptor alpha and beta subunits were differentially expressed in the vestibular periphery of the rat. To begin an understanding of the molecular genetics of these vestibular efferents, this study examined the chromosomal locations of these nicotinic acetylcholine receptor genes in the rat (Rattus norvegicus). Using radiation hybrid mapping and a rat radiation hybrid map server (www.rgd.mcw.edu/RHMAP SERVER/), we determined the chromosomal position for each of these genes. The alpha2-7, alpha9, alpha10, and beta2-4 nicotinic subunits mapped to the following chromosomes: alpha2, chr. 15; alpha3, chr. 8; alpha4, chr. 3; alpha5, chr. 8; alpha6, chr. 16; alpha7, chr. 1; alpha9, chr. 14; alpha10, chr. 7; beta2, chr. 2; beta3, chr. 16; and beta4, chr. 8. With the location for each of these nicotinic subunits known, it is now possible to develop consomic and/or congenic strains of rats that can be used to study the functional genomics of each of these subunits.

Positional cloning of a novel gene on chromosome 16q causing Bardet-Biedl syndrome (BBS2).

Bardet-Biedl syndrome (BBS) is a genetically heterogeneous autosomal recessive disorder with the primary clinical features of obesity, pigmented retinopathy, polydactyly, hypogenitalism, mental retardation and renal anomalies. Associated features of the disorder include diabetes mellitus, hypertension and congenital heart disease. There are six known BBS loci, mapping to chromosomes 2, 3, 11, 15, 16 and 20. The BBS2 locus was initially mapped to an 18 cM interval on chromosome 16q21 with a large inbred Bedouin kindred. Further analysis of the Bedouin population allowed for the fine mapping of this locus to a 2 cM region distal to marker D16S408. Physical mapping and sequence analysis of this region resulted in the identification of a number of known genes and expressed sequence tag clusters. Mutation screening of a novel gene (BBS2) with a wide pattern of tissue expression revealed homozygous mutations in two inbred pedigrees, including the large Bedouin kindred used to initially identify the BBS2 locus. In addition, mutations were found in three of 18 unrelated BBS probands from small nuclear families.

The use of designer rats in the genetic dissection of hypertension.

The rat is a well-established model for hypertension research, in both physiologic and pharmacologic study. Quantitative trait loci (QTL) for blood pressure and related phenotypes have been described on every rat chromosome; therefore, more simplified models must be generated to identify and study the function of the gene(s) located by QTL analysis. Designer rat strains, such as congenic and consomic strains, which share phenotypic and genotypic characteristics with humans but with a greatly simplified genetic background, would yield a powerful platform for functional studies, especially when combined with microarray technologies. Development of these designer rats would result in better-defined disease models that can be used in physiologic and applied pharmacologic studies to better treat human essential hypertension.

Biased over- or under-reporting is characteristic of individuals whether over time or by different assessment methods.

Seven studies with repeated measurements of energy intake and/or nitrogen intake were examined to determine whether misreporting is characteristic of some persons or occurs randomly. Four of the studies were validated by doubly labeled water measurements of energy expenditure. Reporting validity was expressed as the ratio of energy intake to energy expenditure. Ratios were consistently below the expected value of 1.0 for some subjects and consistently above 1.0 for others, indicating characteristic reporting validity within subjects. Two year-long studies provided 4 to 12 measurements and a total number of days sufficient to measure individual habitual intake. Subjects mean energy intake to basal metabolic rate (BMR) ratios were < 1.35 in 45% and 47% and < 1.35 at every measurement in 25% of subjects. This indicated persistent underreporting over time, because 1.35 x BMR is the minimum energy expenditure compatible with a normally active lifestyle. Three of the studies used more than 1 assessment method (validated by doubly labeled water and/or urinary nitrogen excretion). There was a tendency for persons determined to be underreporters by 1 method to be also underreporters when tested by other methods. We conclude that biased over- or underreporting is characteristic of some persons. Thus, repeat measurements do not necessarily provide valid measures of individual intake, extreme intakes may reflect under- and overreporting rather than true low or high intakes, and subjects most prone to reporting bias may be repeatedly misclassified in quantiles of the distribution. This presents a challenge to dietitians nutritionists, and statisticians both for the design of surveys and the handling of flawed data.

High-throughput scanning of the rat genome using interspersed repetitive sequence-PCR markers.

We report the establishment of a hybridization-based marker system for the rat genome based on the PCR amplification of interspersed repetitive sequences (IRS). Overall, 351 IRS markers were mapped within the rat genome. The IRS marker panel consists of 210 nonpolymorphic and 141 polymorphic markers that were screened for presence/absence polymorphism patterns in 38 different rat strains and substrains that are commonly used in biomedical research. The IRS marker panel was demonstrated to be useful for rapid genome screening in experimental rat crosses and high-throughput characterization of large-insert genomic library clones. Information on corresponding YAC clones is made available for this IRS marker set distributed over the whole rat genome. The two existing rat radiation hybrid maps were integrated by placing the IRS markers in both maps. The genetic and physical mapping data presented provide substantial information for ongoing positional cloning projects in the rat.

Critical evaluation of energy intake using the Goldberg cut-off for energy intake:basal metabolic rate. A practical guide to its calculation, use and limitations.

OBJECTIVES: To re-state the principles underlying the Goldberg cut-off for identifying under-reporters of energy intake, re-examine the physiological principles and update the values to be substituted into the equation for calculating the cut-off, and to examine its use and limitations. RESULTS: New values are suggested for each element of the Goldberg equation. The physical activity level (PAL) for comparison with energy intake:basal metabolic rate (EI:BMR) should be selected to reflect the population under study; the PAL value of 1.55 x BMR is not necessarily the value of choice. The suggested value for average within-subject variation in energy intake is 23% (unchanged), but other sources of variation are increased in the light of new data. For within-subject variation in measured and estimated BMR, 4% and 8.5% respectively are suggested (previously 2.5% and 8%), and for total between-subject variation in PAL, the suggested value is 15% (previously 12.5%). The effect of these changes is to widen the confidence limits and reduce the sensitivity of the cut-off. CONCLUSIONS: The Goldberg cut-off can be used to evaluate the mean population bias in reported energy intake, but information on the activity or lifestyle of the population is needed to choose a suitable PAL energy requirement for comparison. Sensitivity for identifying under-reporters at the individual level is limited. In epidemiological studies information on home, leisure and occupational activity is essential in order to assign subjects to low, medium or high PAL levels before calculating the cut-offs. In small studies, it is desirable to measure energy expenditure, or to calculate individual energy requirements, and to compare energy intake directly with energy expenditure.

Validation of dietary intakes measured by diet history against 24 h urinary nitrogen excretion and energy expenditure measured by the doubly-labelled water method in middle-aged women.

A diet history method for estimating energy and N intakes was validated against 24 h urinary N excretion and energy expenditure measured by the doubly-labelled water (DLW) method. Forty-eight women aged 50-65 years were studied over 1 year. Weighed diet records from 4 d and two 24 h urine collections, for measurement of urinary N excretion, were obtained in each of four seasons. At the end of the year, a diet history was obtained, BMR was measured by whole-body calorimetry, and, in sixteen women, total energy expenditure (EE) was measured by DLW. Energy intake (EI) and N intake (NI) were calculated using food tables. Using weighed records and diet history respectively mean NI were 11.21 (SD 2.09) g and 11.47 (SD 2.40) g (NS) and EI were 8.08 (SD 1.54) MJ and 8.20 (SD 1.86) MJ (NS). Mean urine N:NI and EI:BMR values indicated bias to under-reporting by weighed record and diet history techniques in some individuals, but there was no significant difference between these measures at the group level. The Pearson correlation coefficient (r) for urine N v. NI was 0.81 for the weighed record and 0.38 for the diet history. The correlation of EE v. EI was r 0.48 for weighed record and r 0.11 for diet history. In this study the diet history gave the same estimate of mean intake, but the weighed record appeared to perform better in ranking individuals.

Within- and between-subject variation in energy expenditure measured by the doubly-labelled water technique: implications for validating reported dietary energy intake.

OBJECTIVES: To estimate the total (CVt), within-subject (CVw) and between-subject (CVb) variation in free-living energy expenditure as measured by the doubly-labelled water (DLW) technique. To examine the limitation of the DLW measurement of energy expenditure for evaluating reported energy intake. To estimate the probable minimum and maximum 'habitual' energy expenditures for a sustainable lifestyle. DESIGN: Review and analysis of individual data from 25 studies with repeat DLW measurements of energy expenditure (EE). RESULTS: Pooled mean CVw derived from 21 studies was 11.8% for EE and 12.3% for physical activity level (PAL). Multiple regression analysis of CVw in 25 studies found a positive association with time span between measurements. At zero time CVw for EE was 8.2% rising to 9.6% at 13 weeks and 15.4% at 52 weeks. At the same time points CVw for PAL was 9.1%, 10.0% and 13.4% respectively. Pooled mean CVt was 13.0% for EE and 10.7% for PAL. CVb calculated from pooled mean CVt and CVw was 20.6% for EE and 7.2% for PAL. 95% confidence limits of PAL in 11 age-sex groups averaged 1.2 to 2.2. CONCLUSIONS: The analysis supported previous estimates of 8% for within-subject variation in DLW measurements including analytic plus inherent biologic variation. Variation that included changes in weight, season and activity increased with increased time between measurements to about 15% at a time span of 12 months. Confidence limits of agreement between EE and reported energy intake were estimated to range from +/-15% to +/-32%. Estimates of the range of usual EE for normally active persons ranged from 1.3 to 2.2.

The sensitivity and specificity of the Goldberg cut-off for EI:BMR for identifying diet reports of poor validity.

OBJECTIVE: To explore the specificity and sensitivity of the Goldberg cut-off for EI:BMR for identifying diet reports of poor validity as compared with the direct comparison of energy intake with energy expenditure measured by doubly-labelled water. DESIGN: Twenty-two studies with measurements of total energy expenditure by doubly-labelled water (EE), basal metabolic rate (BMR) and energy intake (EI) provided the database (n=429). The ratio EI:EE provided the baseline definition of under- (UR), acceptable- (AR) and over-reporters (OR), respectively EI:EE <0.76, 0.76-1.24 and >1.24. Four strategies for identifying under- and over-reporters using the Goldberg cut-off were explored. Sensitivity of the cut-off was calculated as the proportion of UR correctly identified and specificity as the proportion of non-UR correctly identified. RESULTS: UR, AR and OR (by EI:EE) were 34, 62 and 4% respectively of all subjects. When a single Goldberg cut-off for the physical activity level (PAL) of 1.55 was used, for men and women respectively the sensitivity was 0.50 and 0.52 and the specificity 1. 00 and 0.99. Using a cut-off for higher PAL traded specificity for sensitivity. Using the cut-off for a PAL of 1.95, sensitivity was 0. 76 and 0.85 and the specificity 0.87 and 0.78 for men and women respectively. Using cut-offs for mean age-sex specific PAL did not improve sensitivity. When subjects were assigned to low, medium and high activity levels and cut-offs for three different PALs used, sensitivity improved to 0.74 and 0.67 without loss of specificity (0. 97 and 0.98), for men and women respectively. If activity levels for men were applied to the womens' data, sensitivity improved to 0.72. CONCLUSION: To identify diet reports of poor validity using the Goldberg cut-off for EI:BMR, information is needed on each subject's activity level.

Comparison of the incidence of complications at induction and emergence in infants receiving oral atropine vs no premedication.

We studied 120 patients less than 1 yr of age, allocated randomly to receive atropine 40 micrograms kg-1 orally 1 h before operation (group A) or no premedication (group B). All patients underwent a standardized anaesthetic, including inhalation induction with halothane followed by atracurium 0.5 mg kg-1, tracheal intubation and positive pressure ventilation. Monitoring during anaesthesia included heart rate, arterial oxygen saturation, temperature and airway conditions at induction and emergence. The incidence of a decrease in arterial oxygen saturation to 94% or less at induction and recovery was similar in both groups (30.5% at induction, 39% at extubation in group A; 31% at induction, 41% at extubation in group B). There were significantly more airway complications in group B both at induction and emergence (25% and 49%, respectively, compared with 9% and 25% in group A; P < 0.015). Mean heart rate at induction and in the peroperative period was significantly higher in the group receiving atropine (P < or = 0.001). There was an increased incidence of bradycardia (decrease in heart rate of > or = 20%) at induction in the non-premedicated group (23% in group B compared with 10% in group A), but this was not statistically significant. We conclude that the incidence of airway complications at induction and emergence was reduced by orally administered atropine premedication.

New target regions for human hypertension via comparative genomics.

Models of human disease have long been used to understand the basic pathophysiology of disease and to facilitate the discovery of new therapeutics. However, as long as models have been used there have been debates about the utility of these models and their ability to mimic clinical disease at the phenotypic level. The application of genetic studies to both humans and model systems allows for a new paradigm, whereby a novel comparative genomics strategy combined with phenotypic correlates can be used to bridge between clinical relevance and model utility. This study presents a comparative genomic map for "candidate hypertension loci in humans" based on translating QTLs between rat and human, predicting 26 chromosomal regions in the human genome that are very likely to harbor hypertension genes. The predictive power appears robust, as several of these regions have also been implicated in mouse, suggesting that these regions represent primary targets for the development of SNPs for linkage disequilibrium testing in humans and/or provide a means to select specific models for additional functional studies and the development of new therapeutics.

Intensity of bacteraemia associated with conservative dental procedures in children.

OBJECTIVES: To explore the individual dento-gingival manipulative procedures that together lead to the placement of a restoration and to estimate the associated intensity of bacteraemia. PATIENTS AND METHODS: Healthy children receiving dental treatment under general anaesthesia provided blood samples 30 seconds after one of four dento-gingival manipulative procedures: 1. Placement of rubber dam, 2. Use of the high speed drill, 3. Use of the slow speed drill, and 4. Placement of matrix band and wedge. Blood cultures were processed to give the percentage prevalence of bacteraemia, the intensity of organisms per millilitre of blood and the identity of the organisms cultured. RESULTS: A total of 257 children were recruited to the study. The percentage positive prevalence of blood cultures was baseline--9.3%, rubber dam placement--31.4%, slow drill--12.2%, fast drill--4.3%, and matrix band and wedge--32.1%. The intensity of bacteraemia was baseline--1.2 cfu, rubber dam placement--1,962 cfu, slow drill--0.3 cfu, fast drill--1.9 cfu, matrix band and wedge--4.8 cfu. CONCLUSIONS: These data indicate that dento-gingival manipulative procedures comprising a simple dental restoration can lead to a bacteraemia comparable to that from dental extractions. It is suggested that these data may indicate the need for antibiotic prophylaxis for some aspects of conservative dentistry.

The role of rats in functional genomics.

Two hundred years of physiological and pharmaceutical studies and a decade of transgenic technology and genomic resources have made the laboratory rat a major model for biomedical research.

Molecular cloning, tissue-specific expression, and chromosomal localization of a novel nerve growth factor-regulated G-protein- coupled receptor, nrg-1.

A novel and differentially expressed gene, named nrg-1, was identified by EST expression profiling and subsequently isolated as a 2.2-kb full-length clone from a rat PC12 cell cDNA library. Sequence analysis reveals that nrg-1 encodes a putative seven transmembrane spanning domain protein with structural features characteristic of receptors belonging to the G-protein-coupled receptor gene superfamily. The 400-amino-acid protein encoded by nrg-1 exhibits a high degree of sequence identity (40-44%) to the Edg receptor family; members include Edg-1, Edg-2, Edg-3, Edg-4, and H218. Both Northern analysis andEST expression profiling revealed that whole-tissue distribution of nrg-1 mRNA is restricted, found almost exclusively in brain. Transcripts of nrg-1 could be ubiquitously detected in different regions, with very prominent expression in lower brain regions such as the midbrain, pons,medulla, and spinal cord. In PC12 cells, nerve growth factor induces neuronal differentiation and repressed expression of nrg-1. Two other agents that differentiate PC12 cells, fibroblast growth factor and dibdutyryl cAMP, down-regulated nrg-1 mRNA levels. Epidermal growth factor, and agent that does not induce differentiation, did not repress nrg-1 mRNA levels. In a PC12 cell mutant that is deficient in protein kinase A activity (AB.11), all three differentiating agents were unable to down-regulate nrg-1 mRNA. Hence, protein kinase A appears to be an obligatory cellular component in nrg-1 mRNA regulation. Chromosomal mapping employing a rat somatic cell readiation hybrid panel demonstrated that nrg-1 is linked to marker D8Rat54 and tightly associated with H218 on chromosome 8.