Fecal S100A12 in healthy infants and children.

BACKGROUND AND AIMS: Fecal S100A12 is shown to be a useful noninvasive marker of gut inflammation. However, the studies to date have not characterised the patterns of expression in healthy young children. This study aimed to determine S100A12 levels in infants and children without symptoms of underlying gut disease. METHODS: Stool samples were collected from healthy infants (<12 months) and children without gastrointestinal symptoms. Faecal S100A12 was measured by immunoassay. RESULTS: Fifty-six children were recruited. Serial samples were obtained from seven term infants over the first 6 months of life. Single samples were obtained from 49 healthy children ranging from 0.16 to 13.8 years of age. Median S100A12 levels were 0.5 mg/kg (ranging from 0.39 to 25) in the healthy children, with high values (>10 mg/kg) in five infants only. There was no variation between gender. Median S100A12 levels in healthy infants remained below the established normal cut-off from birth to six months of age. CONCLUSION: S100A12 levels in well infants and children are almost exclusively lower than the standard cut-off. Transiently higher levels may be seen in early infancy. An elevated level of S100A12 in children older than 12 months of age is likely to represent organic gut disease.

Molecular characterization of bacterial colonization in the preterm and term infant's intestine.

OBJECTIVE: To further define patterns of colonising intestinal microflora in newborn infants utilising molecular methods. METHODS: Ten term and 5 preterm (<32 wk) infants born at the Royal Hospital for Women, Sydney, Australia were enrolled in the present study and followed for 6 mo post partum. Serial stools were collected, DNA was extracted and subjected to PCR-Denaturing Gradient Gel Electrophoresis using a range of primers and sequencing. The effect of gestational length, feeding and delivery method was compared to the pattern of bacterial acquisition. RESULTS: Intestinal bacterial diversity was lower in preterm compared with term infants. For term infants, bacterial DNA detection rates were not associated with feeding or delivery method, although Enterobacteria and Clostridia were commonly identified. The detection rate of Bifidobacteria was lower in preterm infants than term infants. Potential pathogens were detected in preterm infant samples. CONCLUSIONS: Preterm infants frequently have aberrant bacterial colonization of the intestine. Further research is now required to determine if this may contribute to adverse health outcomes.

Longer-term outcomes of nutritional management of Crohn's disease in children.

BACKGROUND: While the short-term benefits of exclusive enteral nutrition (EEN) for induction of remission in children with Crohn's disease (CD) are well documented, the longer-term outcomes are less clear. AIM: This retrospective study aimed to ascertain the outcomes for up to 24 months following EEN in a group of children with CD. METHODS: Children treated with EEN as initial therapy for newly diagnosed CD over a 5-year period were identified. Details of disease activity, growth, and drug requirements over the period of follow-up were noted. Outcomes in children managed with EEN were compared to a group of children initially treated with corticosteroids. RESULTS: Over this time period, 31 children were treated with EEN and 26 with corticosteroids. Twenty-six (84 %) of the 31 children treated with EEN entered remission. Children treated with EEN exhibited lower pediatric Crohn's disease activity index (PCDAI) scores at 6 months (p = 0.02) and received lower cumulative doses of steroids over the study period (p < 0.0001) than the group treated with corticosteroids. Height increments over 24 months were greater in the EEN group (p = 0.01). Although the median times to relapse were the same, the EEN group had a lower incidence of relapse in each time interval and survival curve analysis showed lower risk of relapse (p = 0.008). CONCLUSIONS: EEN lead to multiple benefits beyond the initial period of inducing remission for these children, with positive outcomes over 2 years from diagnosis. Of particular clinical relevance to growing children was the reduced exposure to corticosteroids.

Fecal M2-PK in children with Crohn's disease: a preliminary report.

BACKGROUND AND AIMS: Although active inflammatory changes in chronic Crohn's disease (CD) can be detected with serum inflammatory markers, these have low specificity and sensitivity. Stool markers of inflammation, such as M2-pyruvate kinase (M2-PK), permit more direct assessment of mucosal inflammation. The aim of this study was to assess levels of M2-PK in children with active CD and to compare to levels in healthy control children. METHODS: Fecal levels of M2-PK were measured by immunoassay using stored stool samples from children with untreated (active) CD and healthy control children. Correlations between M2-PK levels and disease activity scores and serum inflammatory markers were performed. Comparison was also made between M2PK and a second fecal inflammatory marker, S100A12. RESULTS: Mean fecal M2-PK levels were higher in the 17 patients with active CD than in the 21 healthy controls (p = 0.0007). M2-PK levels did not correlate with disease activity scores or serum inflammatory markers. There was a trend for children with ileocolonic disease to have higher levels of M2-PK in their stool compared to those with colonic disease or isolated ileal disease. Fecal M2PK did not correlate with fecal S100A12 in children with active CD. CONCLUSION: Fecal M2-PK is increased in children with active CD, indicating that this marker may be a useful non-invasive marker for gut inflammation. Further studies of M2PK are required in additional settings with larger cohorts of children with CD and with comparison to other stool markers.

Faecal calprotectin, lactoferrin, M2-pyruvate kinase and S100A12 in severe ulcerative colitis: a prospective multicentre comparison of predicting outcomes and monitoring response.

OBJECTIVE: To compare four faecal markers for their ability to predict steroid refractoriness in severe paediatric ulcerative colitis (UC). Construct validity and responsiveness to change were also assessed. METHODS: This was a prospective multicentre cohort study. Stool samples from 101 children (13.3 + or - 3.6 years; Pediatric UC Activity Index (PUCAI) at admission 72 + or - 12 points) were obtained at the third day of intravenous steroid therapy. Repeated samples at discharge were obtained from 24 children. Predictive validity was assessed using diagnostic utility statistics to predict steroid failure (ie, the need for salvage treatment). Concurrent validity was assessed using correlational analysis with the following constructs: PUCAI, Lindgren and Seo scores, physician's global assessment, albumin, erythrocyte sedimentation rate and C-reactive protein (CRP). Responsiveness was assessed using test utility and correlational strategies. RESULTS: Median values (IQR) were very high at baseline for all four markers (calprotectin 4215 microg/g (2297-8808); lactoferrin 212 microg/g (114-328); M2-pyruvate kinase (M2-PK) 363 U/g (119-3104); and S100A12 469 microg/g (193-1112)). M2-PK was numerically superior to the other three markers and CRP in predicting response to corticosteroid treatment (area under the receiver operating characteristic (ROC) curve 0.75 (95% CI 0.64 to 0.85; p<0.001) vs <0.65 for the others). However, it did not add to the predictive ability of the PUCAI (area under the ROC 0.81 (95% CI 0.73 to 0.89)). M2-PK also had the highest construct validity but with a modest mean correlation with all constructs (r=0.3; p<0.05). None of the markers was responsive to change (Spearman's rho correlation with change in the PUCAI <0.1; p>0.05, area under the ROC curve <0.65; p>0.05). CONCLUSIONS: The four markers were greatly elevated in severe paediatric UC. Only M2-PK had good construct and predictive validity, and none was responsive to change. The PUCAI, a simple clinical index, performed better than the faecal markers in predicting outcome following a course of intravenous corticosteroids in severe UC.

Nutritional status of children with coeliac disease.

AIMS: The main aim of this study was to assess the nutritional status of children with newly diagnosed Coeliac disease (CD)with comparison to matched controls. A further aim was to assess relationships between presentation patterns and nutrition in childhood CD. METHODS: The nutritional status of newly diagnosed CD was assessed by anthropometry, Bioelectrical Impedance and serum leptin levels, and contrasted to age and gender matched controls. RESULTS: Twenty-five children with CD (mean age of 8.2 +/- 4.5 years) and 25 control children (mean age 8.1 +/- 4.4.) were enrolled. Thirteen (52%) children with CD had gastrointestinal symptoms with 14 having a family history of CD. At presentation 8.7% were wasted, 4.2% were stunted and 20.8% overweight, although none were obese. Mean height and weight for age, other nutritional parameters and serum leptin did not differ between the groups. Serum leptin correlated with BMI in both groups. CONCLUSIONS: Children with CD more commonly present with atypical symptoms than with classical features. Variations in nutrition (under to overnutrition) may be seen at diagnosis, without relationship to the presence of symptoms. Leptin levels were not altered specifically in the setting of CD. Nutritional assessment remains important in the assessment and management of CD in children.

Sustained modulation of intestinal bacteria by exclusive enteral nutrition used to treat children with Crohn's disease.

BACKGROUND: The use of exclusive enteral nutrition to treat paediatric Crohn's disease (CD) is widely accepted, although the precise mechanism(s) of action remains speculative. AIM: To investigate the changes to key intestinal bacterial groups of Eubacteria, Bacteroides, Clostridium coccoides, Clostridium leptum and Bifidobacteria, during and after exclusive enteral nutrition treatment for CD in paediatric patients and correlate these changes to disease activity and intestinal inflammation. METHODS: Stool was collected from six children at diagnosis of CD, during exclusive enteral nutrition and 4 months post-therapy, and from seven healthy control children. The diversity of bacteria was assessed by polymerase chain reaction-denaturing gradient gel electrophoresis with changes to bacterial diversity measured by Bray-Curtis similarity, intestinal inflammation assessed by faecal S100A12 and the disease activity assessed by PCDAI. RESULTS: A significantly greater change in intestinal bacterial composition was seen with exclusive enteral nutrition treatment compared with controls. Further, the intestinal bacteria remained altered 4 months following exclusive enteral nutrition completion. Changes in the composition of Bacteroides were associated with reduced disease activity and inflammation. CONCLUSIONS: Exclusive enteral nutrition reduces bacterial diversity and initiates a sustained modulation of all predominant intestinal bacterial groups. Exclusive enteral nutrition may reduce inflammation through modulating intestinal Bacteroides species. The implications of these results for exclusive enteral nutrition therapy and CD pathogenesis should now be the subject of further investigation.