Fungi: Friend or Foe? A Mycobiome Evaluation in Children with Autism and Gastrointestinal Symptoms

Gastrointestinal (GI) symptoms often affect children with autism spectrum disorders (ASD) and GI symptoms have been associated with an abnormal fecal microbiome. There is limited evidence of Candida species being more prevalent in children with ASD. We enrolled 20 children with ASD and GI symptoms (ASD + GI), 10 children with ASD but no GI symptoms (ASD - GI), and 20 from typically developing (TD) children in this pilot study. Fecal mycobiome taxa were analyzed by Internal Transcribed Spacer sequencing. GI symptoms (GI Severity Index [GSI]), behavioral symptoms (Social Responsiveness Scale -2 [SRS-2]), inflammation and fungal immunity (fecal calprotectin and serum dectin-1 [ELISA]) were evaluated. We observed no changes in the abundance of total fungal species (alpha diversity) between groups. Samples with identifiable Candida spp. were present in 4 of 19 (21%) ASD + GI, in 5 of 9 (56%) ASD - GI, and in 4 of 16 (25%) TD children (overall P = 0.18). The presence of Candida spp. did not correlate with behavioral or GI symptoms (P = 0.38, P = 0.5, respectively). Fecal calprotectin was normal in all but one child. Finally, there was no significance in serum dectin-1 levels, suggesting no increased fungal immunity in children with ASD. Our data suggest that fungi are present at normal levels in the stool of children with ASD and are not associated with gut inflammation

Elevated Lipase during Initial Presentation of Ulcerative Colitis in a Pediatric Patient: Do We Check for It

There are very few reports of elevated lipase in pediatric inflammatory bowel disease (IBD). Symptoms of pancreatitis may be masked by abdominal pain in pediatric IBD. During the initial presentation of IBD in our patient, lipase was elevated to more than 3 times the upper limit of normal. Normalization of values coincided with remission of IBD. This may be due to extraintestinal involvement of the pancreas as part of the inflammatory process or due to leakage of pancreatic enzymes from an inflamed gut or mediated by inflammatory cytokines. Checking pancreatic enzymes during initial presentation of IBD may, therefore, be important to determine if pancreatic involvement has resulted from the inflammation in IBD or as an adverse effect of therapy. If unchecked, recurrent subclinical pancreatitis may be masked by IBD symptoms and missed prior to starting IBD therapy. This may result in chronic pancreatic insufficiency as reported in 50% of adults with IBD. Early detection of elevated pancreatic enzymes in IBD may help direct the management strategy, as treatment of the underlying inflammation in IBD may be the most important management for resolution of pancreatitis instead of cessation of therapy for fear of iatrogenic medication-induced pancreatitis

Baseline clinical characteristics of the study groups.

<p>Abbreviations: Body Mass Index (BMI), White blood cell count (WBC), highly sensitive C-reactive protein (hSCRP), Blood urea nitrogen (BUN).</p>*<p>denotes p values obtained from non-parametric Kruskal-Wallis test.</p

Fecal calprotectin at various study points.

<p>Median fecal calprotectin at various study time points are shown. There is a significant difference between calprotectin in LR-treated vs. placebo groups after treatment. Nonetheless all calprotectin values remained in the normal clinical range (fecal calprotectin <160 µg/g).</p

Distribution (frequency, percentage) of the adverse events by study arm and type during treatment phase.

<p>One patient withdrew from study after randomization before receiving treatment.</p><p>None of the adverse events were identified to be related to treatment group by an independent data safety monitoring board (DSMB).</p><p>No significant difference in adverse events between groups (p = 0.46)</p

Baseline research laboratory characteristics of the study groups^*.

<p>Abbreviations: T regulatory cells (Tregs), myeloid dendritic cells (mDC); plasmacytoid dendritic Cells (pDC); interferon-gamma (IFNγ); interleukin (IL); tumor necrosis factor-alpha (TNFα); toll-like receptor (TLR); mean fluorescence intensity (MFI).</p>*<p>Mean, standard deviations with p-values based on t-tests reported for values following a normal distribution. Median, interquartile ranges (IQR) with p-values based on Kruskal-Wallis test reported for values with skewed distribution.</p>**<p>PBMC (1×10⁶) stimulated by PMA (50 ng/ml) and inomycin 1 (µg/ml) for 16 hours.</p

Representative DGGE images.

<p>Representative denaturing gradient gel electrophoresis images from six patients, three treated with LR (Treatment Group LR001, LR011, LR014) and three with placebo (Placebo group LR003, LR009, LR059). Each lane in a gradient gel shows bands of 16S rDNA detected in each of five stool specimens collected at 0, 1, 2, 3 and 6 months. Each band represents a different bacterial 16S rDNA sequence, i.e. a different bacterial taxon. Time 0 stools were collected before patients began consuming LR or placebo. Time 1 and 2 stools were collected after patients had been consuming LR or placebo for one and two months respectively. Time 3 and 6 stools were collected one month and four months after patients had discontinued consuming LR or placebo. For each patient, differences in band patterns across sampling time points indicate changes in the composition of bacteria, over time, in their stools. For example, Patient LR001 exhibited a notable change in stool bacterial composition between Time 0, a stool collected before consuming any LR, and Time 1, a stool collected after consuming LR for one month. Likewise, the placebo treated Patient LR059, exhibited a notable change in stool bacterial composition over the same interval. In contrast LR treated Patient LR011 exhibited no change in band pattern over that interval. We compared the total number of band pattern changes over each interval in the LR treated and placebo treated groups and found no significant difference in the influence of LR treatment compared to placebo treatment on changes in stool bacterial composition over any interval examined.</p