(13)C mixed triglyceride breath testing using infrared spectrometry: comparison of two devices in early infancy.

BACKGROUND/OBJECTIVES: The (13)C mixed triglyceride (MTG) breath test has been proposed for the non-invasive assessment of fat digestion and absorption. To evaluate whether reference values for the adequacy of fat absorption, set in the non-dispersive infrared spectrometry (NDIRS) system software proposed for healthy children and adults using the (13)C MTG breath test, are also applicable to infants of <5 months of age. SUBJECTS/METHODS: (13)C MTG breath testing with the NDIRS technique was performed in 54 healthy infants <5 months of age (38 breast-fed, 16 formula-fed) and six infants diagnosed with cystic fibrosis (CF) using two NDIRS devices, IRIS and FANci2. RESULTS: The IRIS results were slightly higher compared with those assessed by the FANci2 device. The minimum cutoff value for pancreatic sufficiency (PS) is set as a cumulative percentage dose of (13)C recovered (cPDR) after 5 h of 13.0%. Pancreatic function status of six CF infants, three with PS and three with pancreatic insufficiency (PI), according to the 72 h-faecal fat balance test could be correctly determined with the (13)C MTG breath test using two NDIRS techniques. However, if these reference values had been used to determine pancreatic function status in healthy infants, 26 out of 54 infants would have been misclassified as pancreatic insufficient. CONCLUSIONS: Although the (13)C MTG breath test with the MS technique has the potential to be a suitable assessment of fat absorption in infants, the technique of NDIRS appears too insensitive in an infant population group.

Nutritional care in children with cystic fibrosis: are our patients becoming better?

Over the last 30 years, major advances have occurred in our understanding of the disorder cystic fibrosis (CF) with the discoveries of the underlying chloride transport defect and the 'CF gene', the CF transmembrane conductance regulator gene. Equally important from a clinical and patient perspective are the improvements in median survival from less than 10 to 20 years prior to 1980, approaching 30 years during the 1980s and over 45 years more recently. Improved antibiotic regimens and lung clearance therapy contributed to the enhanced survival, but a key factor accredited as adding a further 10 years to the median survival was improving and then maintaining normal growth and nutrition. In the main, the latter were achieved by adherence to a 'high-fat high-energy' diet rather than the advocated virtually universal policy of the 'low fat', which was associated with wasting and linear growth failure. The high-fat diet in conjunction with better control of malabsorption due to microspheric pancreatic enzyme replacement therapy, attention to adequate fat-soluble vitamin supplementation and newborn screening has ensured that at least 80-90% of children with CF will achieve better health and survival through their adult years.

Novel mutations in abetalipoproteinaemia and homozygous familial hypobetalipoproteinaemia.

Abetalipoproteinaemia (ABL) and homozygous familial hypobetalipoproteinaemia (FHBL) are rare inherited disorders associated with low or undetectable levels of apolipoprotein B (apoB)-containing lipoproteins. Patients present with the symptoms and sequelae of fat malabsorption, including fat-soluble vitamin deficiencies. We describe two novel mutations: one an APOB gene mutation causing FHBL and the other a microsomal triglyceride transfer protein (MTP) gene mutation causing ABL. Two siblings of consanguineous parents were homozygous for an apoB mutation 4339delT causing an apoB-30.9 truncation. In another family, a boy born to consanguineous parents was homozygous for a 319 bp in-frame deletion of MTP exon 15 (c.2076-39_2303 + 52del319). All three children presented with malabsorption and liver dysfunction and had similar very low serum lipid, apoB, and fat-soluble vitamin levels. The FHBL parents had low serum lipid and apoB profiles distinguishing the disorder from the normal levels in ABL parents. Future patients presenting with FHBL or ABL should be genotyped to provide further insight into the varying clinical severity related to molecular heterogenicity in these two conditions.

Resting energy expenditure in females with cystic fibrosis: is it affected by puberty?

OBJECTIVE: The aims of this study were to determine the effect of puberty and the menstrual cycle on resting energy expenditure (REE) in females with cystic fibrosis (CF). DESIGN: Cross-sectional study. All participants had measurements of REE, anthropometry and pubertal staging. The measurements in the postmenarche group were carried out both in the follicular and luteal phases of their menstrual cycle. SETTING: CF outpatient clinic at the Children's Hospital at Westmead. SUBJECTS: Fifty-six females with CF and pancreatic insufficiency (13 postmenarche) were recruited from the hospital clinic and 63 controls (21 postmenarche) were recruited through families and friends of hospital staff. RESULTS: Females with CF had a higher REE than controls (111.6+/-12.8% of predicted from controls P<0.001). There was a significant effect of menarche on REE with a decrease in the postmenarche -470 kJ/24 h compared with premenarche after adjustment for fat-free mass, fat mass and group (control or CF). There was no difference in REE between the follicular and luteal phases for either CF or controls. CONCLUSIONS: Females with CF had raised REE that appeared to be independent of menarche. This study implies all females with CF and pancreatic insufficiency may need more intensive dietary management, owing to raised REE, to maintain growth and nutritional status, and possibly improve survival.

NMR measurements of the diffusional permeability of water in cultured colonic epithelial cancer cells.

The water residence time and diffusional water permeability in colonic epithelial T84 cancer cells was measured using (1)H NMR spectroscopy; the values estimated were 35.2+/-2.8 ms and (7.4+/-0.6)x10(-3)cms(-1), respectively. Water permeability was inhibited to approximately 10% of its original value by the mercurial diuretic, p-chloromercuribenzenesulfonate (PCMBS; 1mM), and fully restored by dithiothreitol (DTT; 1mM). The permeability was also inhibited reversibly to approximately 55%, by extracellular glibenclamide (1mM), an inhibitor of some ATP-binding cassette (ABC) transporters, including the cystic fibrosis transmembrane conductance regulator (CFTR). Addition of the phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine (IMBX; 0.1-1mM) and the adenylate cyclase activator, forskolin (0.1-1mM) did not alter water permeability. It is concluded that in T84 cells water diffuses through the membrane lipid bilayer and via channels that are inhibited by PCMBS, including the channels that are known to be inhibited by glibenclamide.

Iron status of young Vietnamese children in Australia.

OBJECTIVE: The aim of this study was to estimate the prevalence of iron deficiency in Vietnamese children living in Australia and to identify risk factors associated with iron deficiency. METHODS: A cohort of healthy term Vietnamese infants, were followed from birth (n = 210) to 18 months (n = 174) with anthropometry, dietary intake and feeding practices measured at seven time points. Socio-demographic data were collected from the parents at the first home visit. At 18 months iron status was examined by full blood count and plasma ferritin concentration in 129/152 (85%) of the eligible children. Iron depletion was defined as a plasma ferritin level < 10 microg/L. Iron deficiency without anaemia was defined as iron depletion plus MCV < 70fl and iron deficiency anaemia was defined as iron deficiency anaemia plus Hb < 110 g/L. RESULTS: The prevalence of iron deficiency was iron depletion 19.4% (95% CI: 13.0%, 27.3%), iron deficiency without anaemia 3.1% (95% CI: 0.9%, 7.8%) and iron deficiency anaemia 3.9% (95% CI: 1.3%, 8.8%). Multiple regression analysis showed three significant predictors of iron deficiency: cows milk intake (negative effect), meat, fish or poultry intake (positive effect) and weight gain (negative effect). A cows milk intake > or = 650 mL/day was a risk factor for iron deficiency. CONCLUSION: Prevalence of iron deficiency at 18 months was high despite appropriate infant feeding practices during the first year. Modification of the diet in the second year of life may decrease the risk of iron deficiency in Vietnamese children.

Growth and feeding practices of Vietnamese infants in Australia.

OBJECTIVE: To investigate the growth and feeding practices in first-generation Vietnamese infants living in Australia. DESIGN: Cohort study. SETTING: The study was conducted between 1999 and 2002 in Sydney. SUBJECTS: A total of 239 Vietnamese women were recruited randomly from antenatal clinics, and of these 210 were initially seen. During the first year, 20 cases (9.5%) were lost to follow-up. Data were collected at 0.5, 2, 4, 6, 9 and 12 months. RESULTS: Vietnamese infants were significantly longer and heavier than reference data (both P<0.0001). The Vietnamese infants had a significant decline in weight growth with age compared with reference data (P<0.001). The Vietnamese infants had marginally higher s.d. score for ideal weight for length than reference data (P=0.044). There was a significant decline in ideal weight for length with age compared with reference data (P=0.0065). Both parents were significantly shorter (mean s.d. height scores: -1.5+/-0.8 (mother) and -1.8+/-0.8 (father)) than reference data (P<0.001). The incidence of breast feeding was 79%, but half of the breast feeding women had stopped breast feeding by 3 months. A total of 162 (79.8%) infants were given infant formula within the first week, of whom 131 (80.1%) were fed infant formula within the first 24 h after birth. CONCLUSIONS: Vietnamese infants in this study had growth comparable with reference data despite their parents being shorter than reference data. Breast feeding duration was short with infant formula being introduced early.

The use of body mass index to predict body composition in children.

BACKGROUND: Simple anthropometric indices of body composition have particular appeal for use in children, and as such body mass index (BMI) has been used to predict percentage body fat in a number of studies. AIM: To evaluate the relationship between BMI and percentage body fat (%body fat) and a proposed, more appropriate relationship between BMI and fat mass/height(2) in a cohort of young children. SUBJECTS AND METHODS: Cross-sectional study of 109 children aged between 6 and 10 years residing in either Sydney or Brisbane, Australia. Weight and height were measured using standard methods. Body composition was measured using a stable isotope method to firstly determine total body water and subsequently fat free mass. RESULTS: The correlation between BMI and fat mass/height(2) was markedly greater than that between BMI and percentage body fat. In the entire group of children the R(2) (x100%) value for the relationship between BMI and fat mass/height(2) was 73.3% compared with 46.5% for the relationship between BMI and percentage body fat. CONCLUSIONS: We have shown that the use of BMI to predict fat mass/height(2), and consequently percentage body fat, is superior to the use of BMI to predict percentage body fat based directly upon the R(2) values of the above analysis.

Comparison of total energy expenditure and energy intake in children aged 6-9 y.

BACKGROUND: The accurate measurement of food intake in children is important for assessing nutritional status. OBJECTIVE: We sought to both compare measurements of energy intake (EI) from diet records and of total energy expenditure (TEE) by the doubly labeled water (DLW) method and to investigate misreporting of EI. DESIGN: Forty-seven children (22 boys and 25 girls) aged 7.4 +/- 0.8 y ( +/- SD) were recruited from 25 schools in western Sydney. TEE was measured by DLW over 10 d and EI by use of 3-d food records. Misreporting was defined as [(EI - TEE)/TEE] x 100%. RESULTS: Girls had a higher (P = 0.02) percentage of body fat (28.2 +/- 7.0%) than did boys (22.9 +/- 8.0%); otherwise there were no differences among sex. Although mean (+/-SD) values for EI (7514 +/- 1260 kJ/d) and TEE (7396 +/- 1281 kJ/d) were not significantly different, there was no significant correlation between EI and TEE. EI and TEE were 9% and 11% lower, respectively, than current World Health Organization recommendations for EI. The relative bias (mean difference, EI - TEE) was low at 118 kJ/d, but the limits of agreement (bias +/- 2 SD of the difference) were wide at 118 +/- 3345 kJ/d. Although the mean percentage of misreporting was low (4 +/- 23%), the high SD indicates large intraindividual differences between EI and TEE. The most significant predictor of misreporting was dietary fat intake (r(2) = 0.45, P < 0.0001). Misreporting was not associated with sex or body composition. CONCLUSIONS: In this age group, reported EI is not representative of TEE at the individual level. However, at the population level, 3-d food records may be used for surveys of EI by 6-9-y-old children.

Variations in the measurement of resting energy expenditure in children with cystic fibrosis.

OBJECTIVE: To investigate in children with cystic fibrosis (CF) and children without CF: (1) the test-retest reproducibility of a 20 min resting energy expenditure (REE) measurement; and (2) the long-term reproducibility of REE measurements in children with CF using longitudinal data. DESIGN: Cross-sectional study and longitudinal cohort. SETTING: A tertiary referral paediatric hospital. SUBJECTS: A total of 31 (11 male, 20 female) children (aged 12.8+/-3.6 y) with CF and 32 (14 male, 18 female) healthy children without CF (aged 12.2+/-2.3 y) were enrolled in the short-term reproducibility study. Long-term REE measurement reproducibility was assessed in another 14 children (5 male, 9 female) with CF, comparing their initial REE measurement with a subsequent measurement 1-2 y later. METHODS: All children had measurements of height, weight, skinfold thickness and indirect calorimetry. RESULTS: There was no statistically significant difference in REE between repeated measurements in children with CF (mean+/-s.d., 6240+/-1280 and 6220+/-1315 kJ/24 h) and in the children without CF (6040+/-956 and 6015+/-943 kJ/24 h). For the children with CF, the intraclass correlation coefficient was 0.99 and for children without CF the intraclass correlation coefficient was 0.97. The measurement errors were 119 and 177 kJ, respectively. Approximately 80% of the variation in REE in the CF group and 70% in the group without CF was explained by fat-free mass (FFM). Analysis of the longitudinal CF data show there was no difference in REE between a child's first measurement (5140+/-1140 kJ) and their subsequent measurement (5460+/-1190 kJ), after adjustment for changes in body size between the measurements. CONCLUSION: This study has demonstrated that a short-term 20 min REE measurement is reproducible and therefore valid in children with CF and children without CF. These results also indicate that in children with CF, long-term REE measurements are reproducible.

Total energy expenditure, body fatness, and physical activity in children aged 6-9 y.

BACKGROUND: The recent worldwide increase in the prevalence of childhood obesity may be due in part to a decrease in children's physical activity levels. OBJECTIVE: The current study of children in the years just before puberty aimed to 1) measure total energy expenditure (TEE) by use of the doubly labeled water (DLW) method, 2) determine the proportion of TEE related to physical activity, 3) investigate the relations between measures of physical activity and body fatness, and 4) investigate possible sex differences in these relations. DESIGN: The DLW technique was used to measure TEE over 10 d in 106 healthy children (52 boys) aged 7.8 +/- 0.9 y (x +/- SD). Fat-free mass, and hence fat mass, was derived from the (18)O dilution space. Resting energy expenditure (REE) was calculated with use of the Schofield equations. Physical activity level was calculated as TEE/REE. RESULTS: Mean TEE in both boys (7871 +/- 1135 kJ/d) and girls (7512 +/- 1195 kJ/d) was significantly different (P < 0.0001) from FAO/WHO/UNU recommendations (13% and 9% lower, respectively). There was no significant difference in physical activity level between boys (1.69 +/- 0.22) and girls (1.71 +/- 0.23). In boys but not girls, physical activity level was inversely correlated with BMI (r = -0.37, P < 0.01), fat mass (r = -0.46, P < 0.005), and percentage of body fat (r = -0.50, P < 0.0001). CONCLUSIONS: In boys but not girls, percentage of body fat is inversely associated with physical activity level. Physical activity is one factor contributing to body fatness in boys, but additional factors may influence the size of the fat stores in girls.

The changing pattern of diagnosis of infantile cholestasis.

OBJECTIVE: Cholestatic liver disease in infancy is caused by a wide range of conditions. This study reviews the pattern of diagnosis of infants with cholestasis presenting to a tertiary referral paediatric hospital in Sydney, Australia, during a 12-year period (1985-96). METHODOLOGY: Infants aged less than 6 months with cholestasis were identified retrospectively from hospital records and data retrieved from the medical records. RESULTS: There were 205 infants identified as having cholestatic liver disease. The aetiology of the cholestasis was idiopathic in 25%, metabolic/genetic in 23%, and due to obstruction in 20%, parenteral nutrition in 20%, infection in 9% and bile duct hypoplasia in 3%. CONCLUSIONS: This study highlights the changing patterns of diagnosis of cholestatic liver disease in infants at a tertiary paediatric facility, demonstrating that up to 50% of cases are now due to genetic/metabolic diseases or parenteral nutrition, and a high proportion are due to idiopathic disease.

Parallel secretion of pancreatic phospholipase A(2), phospholipase A(1), lipase, and colipase in children with exocrine pancreatic dysfunction.

The cosecretion of pancreatic lipase and colipase are important in normal fat digestion. As adsorption of phosphatidylcholine to the lipid substrate interferes with lipase activity, hydrolysis to lysophosphatidylcholine with subsequent desorption is also essential for fat digestion. There are some data regarding the secretion of pancreatic phospholipases in normal adults but none in children or patients with pancreatic disease. In the present study, we aimed a) to develop an accurate fast assay method to measure phospholipase A(2) and b) to determine the secretion rate of pancreatic phospholipase A(2) and whether it is cosecreted with lipase and colipase in children with exocrine pancreatic dysfunction. Nine male patients aged 0.5 to 16 y (seven with cystic fibrosis, two with malabsorption) underwent pancreatic stimulation tests. Their colipase and lipase secretion rates were measured by titrimetric methods and phospholipase A(2) and A(1) by phosphorus magnetic resonance spectroscopy ((31)P NMR). It was found that the phospholipases, colipase, and lipase were absent in the two patients with pancreatic insufficiency. In patients with normal absorption, there were marked inter-and intrasubject variations of lipase, colipase, and phospholipase secretion rates that were consistent with the degree of exocrine pancreatic dysfunction. However, in the three 20-min stimulation periods of the pancreatic function test, pancreatic phospholipase is cosecreted with lipase and colipase, and average colipase and phospholipase A(2) secretion rates follow a similar or parallel pattern. These findings are consistent with the important role of pancreatic phospholipases in intestinal phospholipid hydrolysis leading to the desorption of phospholipids from the lipid substrate and enhancing lipid hydrolysis and phospholipid absorption.

Mutation analysis and embryonic expression of the HLXB9 Currarino syndrome gene.

The HLXB9 homeobox gene was recently identified as a locus for autosomal dominant Currarino syndrome, also known as hereditary sacral agenesis (HSA). This gene specifies a 403-amino acid protein containing a homeodomain preceded by a very highly conserved 82-amino acid domain of unknown function; the remainder of the protein is not well conserved. Here we report an extensive mutation survey that has identified mutations in the HLXB9 gene in 20 of 21 patients tested with familial Currarino syndrome. Mutations were also detected in two of seven sporadic Currarino syndrome patients; the remainder could be explained by undetected mosaicism for an HLXB9 mutation or by genetic heterogeneity in the sporadic patients. Of the mutations identified in the 22 index patients, 19 were intragenic and included 11 mutations that could lead to the introduction of a premature termination codon. The other eight mutations were missense mutations that were significantly clustered in the homeodomain, resulting, in each patient, in nonconservative substitution of a highly conserved amino acid. All of the intragenic mutations were associated with comparable phenotypes. The only genotype-phenotype correlation appeared to be the occurrence of developmental delay in the case of three patients with microdeletions. HLXB9 expression was analyzed during early human development in a period spanning Carnegie stages 12-21. Signal was detected in the basal plate of the spinal cord and hindbrain and in the pharynx, esophagus, stomach, and pancreas. Significant spatial and temporal expression differences were evident when compared with expression of the mouse Hlxb9 gene, which may partly explain the significant human-mouse differences in mutant phenotype.

Clinical outcomes of newborn screening for cystic fibrosis.

AIM: To determine how early diagnosis of cystic fibrosis, using neonatal screening, affects long term clinical outcome. METHODS: Fifty seven children with cystic fibrosis born before neonatal screening was introduced (1978 to mid 1981) and a further 60 children born during the first three years of the programme (mid 1981 to 1984), were followed up to the age of 10. The cohorts were compared on measures of clinical outcome, including height, weight, lung function tests, chest x-ray picture and Shwachman score. RESULTS: Age and sex adjusted standard deviation scores (SDS) for height and weight were consistently higher in children screened for cystic fibrosis than in those born before screening. At 10 years of age, average differences in SDS between groups were 0.4 (95% CI -0.1, 0.8) for weight and 0.3 (95% CI -0.1, 0.7) for height. This translates to an average difference of about 2.7 cm in height and 1.7 kg in weight. Mean FEV1 and FVC (as percentage predicted) were significantly higher in the screened cohort at 5 and 10 years of age, with an average difference of 9.4% FEV1 (95% CI 0.8, 17.9) and 8.4% FVC (95% CI 1.8, 15.0) at 10 years. Chest x-ray scores were not different between the groups at any age, but by 10 years screened patients scored an average 5.3 (95% CI 1.2, 9.4) points higher on the Shwachman score. CONCLUSION: Although not a randomised trial, this long term observational study indicates that early treatment made possible by neonatal screening may be important in determining subsequent clinical outcomes for children with cystic fibrosis. For countries contemplating the introduction of neonatal screening for cystic fibrosis, its introduction to some areas in a cluster randomised design will permit validation of studies performed to date.

Phospholipid changes in children with pancreatic sufficiency and insufficiency.

The phospholipid secretion rates and phospholipase A1 and phospholipase A2 activities in biliary-pancreatic secretions of patients with pancreatic sufficiency and insufficiency were measured using 31P-NMR spectroscopy. It was possible to quantify conveniently the individual phospholipids without prior extraction of lipids or treatment of the samples with detergent. The reciprocal nature of the decrease in phosphatidylcholine concentration, compared with the increase in the concentration of 1-lysophosphatidylcholine and 2-lysophosphatidylcholine, suggested a substrate/product relationship consistent with the activities of phospholipase A1 and phospholipase A2, respectively. Although the secretion rates of total biliary phospholipids among the patients with pancreatic sufficiency were similar, the phospholipase A1 and phospholipase A2 activities varied considerably. The latter differences were similar to their pancreatic lipase and colipase secretion levels and hence their degree of pancreatic dysfunction. The biliary-pancreatic secretions from patients with pancreatic insufficiency showed no enzyme activities. Total biliary phospholipid secretions in patients with pancreatic insufficiency with common bile duct stenosis were significantly lower than those in patients with pancreatic sufficiency, and pancreatic insufficiency without common bile duct stenosis.

Accidental paracetamol overdosing and fulminant hepatic failure in children.

OBJECTIVE: To delineate clinical characteristics useful for identifying children with liver failure due to accidental paracetamol overdose. DESIGN: Retrospective review of medical records of all patients admitted from 1985 to 1998 with fulminant hepatic failure. SETTING: Royal Alexandra Hospital for Children, a tertiary referral centre for paediatric liver transplantation. MAIN OUTCOME MEASURES: Contribution of paracetamol to liver failure; other risk factors for liver failure; comparison of clinical features of paracetamol group and others. RESULTS: 18 patients were identified. Eight were considered to have accidental paracetamol hepatotoxicity. In a further three, other risk factors were present but paracetamol was considered a major contributor to liver failure. The seven remaining patients had other risk factors for liver failure. Patients with paracetamol-induced liver failure usually had an acute prodromal illness with prolonged fasting and, at presentation, had encephalopathy, coagulopathy, very high transaminase levels, but disproportionately low total bilirubin levels. Five patients had hypoglycaemia. End-stage liver failure occurred in 4/11 of the paracetamol group compared with 7/7 of the others. CONCLUSION: Accidental paracetamol overdose is associated with fulminant hepatic failure in infants and children. Patients present with high transaminase levels and liver synthetic failure out of proportion to the level of serum bilirubin. Prompt identification of such patients is important as many recover with supportive therapy.

Nutritional intervention and growth in children with chronic renal failure.

OBJECTIVE: To assess whether improving energy intake by tube feeding could prevent growth failure and improve growth rates in children with congenital renal failure. DESIGN: Prospective descriptive study. SETTING: Renal Units, Royal Alexandra Hospital for Children, and Westmead Hospitals. PATIENTS: All children with advanced chronic renal disease (glomerular filtration rate < 30 mL/min/1.73 m2) between 1992 and 1994. INTERVENTION: Tube feeding was commenced if height or weight standard deviation score (SDS) was below the normal range (> -2 SDS) or when height SDS was decreasing and oral intake was not meeting energy requirements. Energy requirements were calculated for median weight for chronological age and sex to provide for catch-up growth. MAIN OUTCOME MEASURES: Growth rate was measured by comparing height and weight SDS at the beginning and end of the study period. Normal growth rate is defined as no change in SDS over time, whereas catch-up growth is defined as an increase in SDS over time. RESULTS: Seven children, mean age 0.6 +/- 0.7 years, with advanced renal failure (mean glomerular filtration rate = 17 mL/min/1.73 m2) caused by congenital renal hypoplasia/dysplasia were studied. All subjects were eventually tube fed for a mean time of 18. 6 +/- 4.5 months. There was no significant change in height SDS (-0. 9 to -1.1) or weight SDS (-0.4 to -0.2). CONCLUSION: Optimizing nutritional intake by tube feeding children with advanced chronic renal failure from an early age resulted in no decline in growth rate; however, catch-up growth was not achieved.