Presence of Segmented Filamentous Bacteria in Human Children and Its Potential Role in the Modulation of Human Gut Immunity.

Segmented filamentous bacteria (SFB) are commensal organisms that grow by anchoring a specialized holdfast structure to the intestinal walls of a variety of animals. Interaction of SFB with Peyer's patches in mice promotes the post-natal maturation of the immune system. We previously reported that the colonization of SFB in humans mainly occurs by 36 months of age, and is difficult to be detected afterward. In this study, we measured the level of SFB in intestinal fluids of human children. SFB were found via qPCR to represent a small fraction of the whole SFB-positive microbiota (105 SFB in 1011 total bacteria). Bacteria with filamentous segmented morphology were observed in intestinal fluids via fluorescent in situ hybridization, and from gut biopsies via scanning electron microscopy. SFB-specific DNA and peptide fragments were also identified via multiple displacement amplification PCR and mass spectrometry. There was an overall positive correlation between the presence of SFB and the titer of total secretory immunoglobulin A (sIgA), which is more apparent in intestinal fluids of the age group of 8-36 months. Afterward there was a decline of SFB in numbers correlated with a reduction of total sIgA. RT-qPCR analysis of the terminal ileal biopsies revealed that the expression of Th17 pathway genes were induced in SFB-positive samples, while the markers of T and B cell receptor signaling pathways were also upregulated. Collectively, these data suggest that SFB is a rare member of microbiota, and may play an important role in the development of human gut immunity.

Colon cleansing protocol in children: research conditions vs. clinical practice.

BACKGROUND AND STUDY AIMS: Colon preparation rates are the limiting factor for a successful diagnostic colonoscopy in children. Different colon cleansing protocols have been published for use in children. Unfortunately, the applicability of those published research protocols has not been formally evaluated in routine clinical practice. We investigated the success rate of our previously published colon cleansing protocol as utilized in our clinical practice. PATIENTS AND METHODS: This was a retrospective study. In the clinical practice, the colon cleansing protocol included PEG-3350 at a dose of 2 g/kg/day plus Dulcolax (Bisacodyl, Boehringer Ingelheim, TX USA) 5 mg/day for 2 days. Adequate colon preparation was graded between 1 - 5, as previously described, and grade ≥ 4.0 was considered an adequate preparation. Patients were instructed to complete a questionnaire that included PEG-3350 dose, number of stools per day, consistency of each stool, and side effects (vomiting, abdominal pain). Clinical and endoscopic results were compared between the protocol under research conditions and routine practice. RESULTS: The success rate of the colon preparation in our clinical practice was similar to the results observed under our research protocol (75 % vs. 73.6 %). Moreover, the total number of stools, stool consistency, and the intubation rate of the terminal ileum were also similar. We concluded, that in our experience, the colon cleansing protocol used under research conditions was effective and appropriate for use in routine clinical practice. CONCLUSION: We recommend testing each new protocol under the routine conditions of clinical practice to confirm its applicability for general practitioners.

Joint ESPGHAN/NASPGHAN Guidelines for the Management of Helicobacter pylori in Children and Adolescents (Update 2016).

BACKGROUND: Because of the changing epidemiology of Helicobacter pylori infection and low efficacy of currently recommended therapies, an update of the European Society for Paediatric Gastroenterology Hepatology and Nutrition/North American Society for Pediatric Gastroenterology, Hepatology and Nutrition recommendations for the diagnosis and management of H pylori infection in children and adolescents is required. METHODS: A systematic review of the literature (time period: 2009-2014) was performed. Representatives of both societies evaluated the quality of evidence using GRADE (Grading of Recommendation Assessment, Development, and Evaluation) to formulate recommendations, which were voted upon and finalized using a Delphi process and face-to-face meeting. RESULTS: The consensus group recommended that invasive diagnostic testing for H pylori be performed only when treatment will be offered if tests are positive. To reach the aim of a 90% eradication rate with initial therapy, antibiotics should be tailored according to susceptibility testing. Therapy should be administered for 14 days, emphasizing strict adherence. Clarithromycin-containing regimens should be restricted to children infected with susceptible strains. When antibiotic susceptibility profiles are not known, high-dose triple therapy with proton pump inhibitor, amoxicillin, and metronidazole for 14 days or bismuth-based quadruple therapy is recommended. Success of therapy should be monitored after 4 to 8 weeks by reliable noninvasive tests. CONCLUSIONS: The primary goal of clinical investigation is to identify the cause of upper gastrointestinal symptoms rather than H pylori infection. Therefore, we recommend against a test and treat strategy. Decreasing eradication rates with previously recommended treatments call for changes to first-line therapies and broader availability of culture or molecular-based testing to tailor treatment to the individual child.

Tissue Transglutaminase Levels Are Not Sufficient to Diagnose Celiac Disease in North American Practices Without Intestinal Biopsies.

BACKGROUND: Celiac serology is crucial for the diagnosis of celiac disease in children. The American guideline for celiac disease in children suggested that positive serology should be followed by confirmatory intestinal histology. The relationship between high tissue transglutaminase titers and celiac disease in children has not been well investigated in children from North America. AIMS: In the present study, we investigated whether different tissue transglutaminase titers in symptomatic children could predict celiac disease without the confirmation of intestinal histology. METHODS: Data from biopsy confirmed celiac children were collected from four different clinics in North America. Clinical, serological, histological, and follow-up data were collected. The accuracy rates of various tissue transglutaminase titers to predict celiac disease in children were calculated. RESULTS: The data from 240 children were calculated. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy rate of tissue transglutaminase titers at ≥10× upper limit of normal were 75.4, 48.8, 87.7, 29.0, and 70.8 %, respectively. Similar data were noted in the other tissue transglutaminase titers (≥3× upper limit of normal, >100 U/ml, or >100 U/ml and >10× upper limit of normal). CONCLUSIONS: The positive predictive value of tissue transglutaminase titers at ≥3× upper limit of normal or higher was too low to predict celiac disease in children. Our data suggested that in routine clinical practice, high titers of tissue transglutaminase are not sufficient to diagnose celiac disease in North American children without intestinal biopsies.

Role of Serum Biomarkers in Early Detection of Non-Alcoholic Steatohepatitis and Fibrosis in West Virginian Children.

BACKGROUND: Obesity, an epidemic among West Virginia children, as well as insulin resistance (IR), is well-established contributors to nonalcoholic steatohepatitis (NASH). Progression of NASH can lead to hepatic fibrosis and cirrhosis, making early detection imperative. The standard for diagnosing NASH is histologically via liver biopsy, which is highly invasive and generally contraindicated in children. By studying serum biomarkers associated with NASH, we aim to identify high risk children who can benefit from a less invasive, alternative approach to the early detection of NASH. METHODS: Seventy one children were prospectively recruited and divided into 3 groups: normal weight without IR (control), obese without IR, and obese with IR. Serum samples were drawn for each patient and biomarker levels were assessed via ELISA kits. RESULTS: Obese without IR and obese with IR patients had significantly elevated levels of lipid metabolism and accumulation markers (FGF-21, NEFA, FATP5, ApoB), oxidative stress markers (dysfunctional HDL, 8-Isoprostane), inflammatory markers(dysfunctional HDL, CK-18) and apoptosis markers (CK-18) compared to control patients (p<0.02). Bilirubin (an antioxidant) was significantly decreased in the obese without IR and obese with IR patients compared to control (p<0.02). CONCLUSION: This study showed a correlation between obesity, IR, and biomarkers associated with NASH in pediatrics patients from West Virginia, with obese with IR patients showing the strongest correlation. These findings support the clinical application of these serum biomarkers as a less invasive method for early detection of NASH and hepatic fibrosis.

Association between retinol binding protein 4 with atherosclerotic markers in obese children.

BACKGROUND: Childhood obesity increases cardiovascular risk during adulthood. Retinol-binding protein-4, a pro-inflammatory adipokine, associated with obesity and insulin resistance also plays a role in atherogenesis in adults. The goal of this study was to identify the relation between RBP4 and atherogenic markers in obese children. METHODS: In a cross-sectional study, obese and non-obese children (8-18 years) were prospectively recruited from a pediatric Appalachian population. Clinical markers such as lipid profile, HbA1c, markers of insulin resistance and plasma levels of RBP4, sVCAM-1 and oxidized-low-density lipoprotein (Ox-LDL) were measured. RESULTS: Compared to non-obese children, RBP4 (P=0.016) and Ox-LDL (P<0.001) were significantly higher in obese children and were positively correlated with Body Mass Index (P<0.001), BMI-SDS (Standard-Deviation Score) (P<0.001) and waist circumference (P=0.03). CONCLUSIONS: No significant correlation was found between inflammatory markers and Homeostatic Model Assessment-2, HDL, triglycerides, and HbA1c in obese children.

Rate of celiac disease in children: view from the endoscopy suite.

OBJECTIVES: The low rate of celiac disease diagnosed in children from the United States may be limited by the practice of "serology-led" diagnosis. The frequency of seronegative celiac disease is unknown, but is underestimated in children and may result in misdiagnosis of celiac disease. The aim of the present study was to investigate the rate of celiac disease after upper endoscopy (esophagogastroduodenescopy [EGD]) with no prior positive celiac serology compared with the rate of celiac disease followed by positive serology. METHODS: Charts of all of the first diagnostic EGDs in children (2009-2013) were retrospectively reviewed. Patients with confirmed celiac disease were divided into 4 groups: group A, positive EGD/positive serology (histology-led diagnosis); group B, positive serology/positive histology (serology-led diagnosis); group C, positive histology followed by negative serology (control 1); and group D, positive serology followed by negative histology (control 2). RESULTS: A total of 761 upper endoscopic charts were reviewed. Of these, 15 children were confirmed with celiac disease (1.97%). There was no significant difference in the demographic data or clinical symptoms between group A and group B. No significant difference was observed in the rate of celiac disease between histology-led celiac diagnosis (group A) and serology-led celiac diagnosis (group B) (1.18% vs 0.79%, P = 0.273). CONCLUSIONS: The rate of celiac disease in endoscopy-led diagnosis was comparable to that in the serology-led diagnosis, suggesting that to increase the detection of celiac disease in children, an adequate number of intestinal biopsies should be performed in every diagnostic upper endoscopic procedure.

Safety of infliximab in children with IBD: the experience of an academic center in WV.

BACKGROUND: The immune-modulating drug, infliximab, is approved for Inflammatory Bowel Disease (IBD) treatment in children. Chronic therapy with infliximab is associated with the development of early and delayed infusion reactions. We reviewed our experience with infliximab treatment and its side effects in a cohort of children diagnosed with IBD who were treated in our clinic. METHODS: A retrospective analysis of all IBD children treated with infliximab in our center from 2006-2011 was performed. The demographic, chronological and clinical data were recorded. The infiximab infusion was given at 5mg/ kg according to a standard protocol after pre-treatment with low dose steroid and diphenhydramine. RESULTS: Of 30 IBD patients (23 CD/7 UC) receiving 454 infusions (341 CD/113 UC), six (20%) patients experienced early infusion reactions. Two (6.7%) patients had a delayed reaction; of those, both required intestinal resection. CONCLUSION: Our study is the first to address the safety of infliximab infusion reactions in children in the state of WV. Our results lend support to the use and safety of infliximab in children with moderate to severe IBD.

Does Helicobacter pylori protect against eosinophilic esophagitis in children?

BACKGROUND: Helicobacter pylori infection and eosinophilic esophagitis (EoE) in children seem to have a reversed association with socioeconomic status (hygienic condition) and allergy conditions. While Hp infection (Hp) is highly associated with poor hygiene and/or poor socioeconomic status, but not with allergic conditions (asthma, rhinitis, etc.), EoE has the opposite epidemiological relationship (high association with allergy but low with low hygienic conditions). AIM: To investigate the association between Hp infection and EoE in children. METHODS: A retrospective chart review of all children who undergo the first upper endoscopy procedure in the gastroenterology clinic, between 2007 and 2012, was performed. Demographic, endoscopic and histological data were collected. The data was divided into 4 diagnostic groups: Hp infection, EoE, reflux esophagitis, and children who had normal histology. The relationship between Hp positive children and the other groups was performed. RESULTS: A total of 966 charts were available for review. Esophagitis, idiopathic gastritis, EoE, and Hp infection were detected in 268 (28%), 480 (49%), 62 (6%), and 31 (3%) children, respectively. The mean age of the EoE group was significantly lower compared to all reference groups (p < .002), but no significant different was detected among the reference groups (gastritis, GERD, and Hp infection; p = 1.00). Simple logistic regression analysis using Hp infection as a predictor for EoE did not find a significant relationship between these two variables (p-value = .471, OR = 0.478, 95% CI 0.06-3.56). However, multivariable logistic regression analysis between EoE and the reference groups indicated a significant negative relationship between Hp infection and EoE (p-value = .023, adjusted OR = 0.096, 95%CI 0.013-0.72). Neither gastritis nor GER showed significant relationship with EoE (p-values are 1.000 and .992, respectively). CONCLUSION: A reversed association between Hp and EoE was found in a cohort of West Virginia children. The possible explanations for these findings are discussed.

Upper endoscopy in children: do symptoms predict positive findings?

Upper endoscopy (esophagogastroduodenoscopy or EGD) is an important diagnostic tool for many gastrointestinal symptoms. In recent years, the number of EGDs has increased dramatically. Unfortunately, the rate of negative (normal) EGD in children is high, approximating 50% of all procedures. To decrease the cost of EGD procedures, it is important to assess which clinical symptom would detect positive findings. This information may also be valuable in improving the referral practices of the primary care physicians for EGD. In a retrospective study, we investigated the pathological yield of the first EGD in children referred for various symptoms. Abdominal pain was the most common referral symptom and the best predictor of positive EGD, reaching an accuracy level of 79.9%. All other investigated symptoms had less than 50% accuracy. We concluded that most gastrointestinal symptoms in children have a poor predictive value for positive EGD. A cost-benefit analysis of EGD in children is needed.

The diagnostic yield of upper endoscopy procedures in children- is it cost effective?

Upper endoscopy is an invaluable tool for the diagnosis and treatment of various gastrointestinal symptoms in children. Over the years, the number of endoscopic procedures performed in different medical centers has increased considerably and the cost associated with the procedure has become unsustainable. Recently, the US government has investigated this topic and has suggested steps to reduce the cost and use of endoscopic procedures in the adult population, changes that have not been accepted favorably by the American Gastroenterology Associations (AGA). In the present report, we evaluate the diagnostic yield of the procedure in children and suggest steps to reduce the annual number of upper endoscopic procedures in children. The diagnostic yield and the cost-effectiveness of the procedure are also discussed.

Etiology and management of esophageal impaction in children: A review of 11 years.

INTRODUCTION: Esophageal impaction in children is a medical emergency that needs an immediate medical attention. In the last years multiple objects have been detected and were removed endoscopically. In the last decade, Eosinophilic Esophagitis (EoE) has been associated with esophageal stenosis, leading to esophageal impaction with various objects including toys, food, coins and other. AIM: To determine the various objects and symptoms associated with impactions and to develop algorithm that guides Emergency Department Physicians and Primary care Providers when facing such problem in pediatric patient. MATERIAL AND METHODS: A retrospective chart review of all children referred to the pediatric gastroenterology service at Marshall University for FB impaction between 2000 and 2010 was performed. Foreign bodies treated by other specialties were excluded from this review. Clinical and demographic data were retrieved. RESULTS: A total of 291 charts with the diagnosis of foreign body reviewed of which only 126 met the inclusion criteria and considered for final calculation. Male/Female ratio and mean age were 1.17:1.0, and 3.65 ± 0.75 years, respectively. Time of impaction before ER evaluation was: < 24hrs in 75 patients, >24hrs in 17 and in 34 the time of ingestion was not known. Radiology was performed in 118 patients, of whom 1 had pneumo-mediastinum and 1 had pneumothorax. Endoscopic localization of the objects showed: 81- proximal, 23- middle esophagus, and 22- in the distal esophagus. The esophageal mucosa showed non-specific changes in 37 patients, 5 with EoE features, 1 patient with hiatal hernia, 1 patient with candidiasis, and in 5 pts had no mucosal description. CONCLUSION: Children with foreign body in the esophagus are commonly seen in our Hospital ER. Those children may harbor undiagnosed medical conditions that will need future therapy. Careful endoscopic assessment with mucosal biopsies is needed to diagnose those conditions earlier. Referral to a GI specialist may be warranted.

Immunoreactivity of p53 and Ki-67 for dysplastic changes in children with eosinophilic esophagitis.

Eosinophilic esophagitis (EoE) has been recognized as a chronic disease of the esophagus with significant involvement of the mucosal immune system. Similar conditions in other gastrointestinal diseases have led to a malignant transformation (ie, inflammatory bowel disease, celiac disease, etc). MIB-1 (Ki-67) and p53 are monoclonal antibodies that are used to detect early markers for dysplastic changes, possibly leading to cancer development in adult patients with chronic gastroesophageal reflux disease (GERD), Barrett's esophagus, and/or the adenocarcinoma sequence of the esophagus. Only limited studies of these cell markers have been published in children with EoE. The aim of this study is to examine p53 and Ki-67 cell markers in children with EoE before and after medical therapy. The immunohistochemical staining of cell markers p53 and Ki-67 was examined in esophageal biopsies of children diagnosed with EoE, GERD, and normal esophagus. In addition, biopsies from adults with EoE and adenocarcinoma were used as positive controls. In 10 children who were successfully treated for EoE, immunohistochemical staining was compared before and after medical therapy. The immunohistochemical staining of p53 and Ki-67 was increased in children with EoE compared with children with a normal esophagus but not in children with GERD. Children with EoE posttherapy had significantly lower immunohistochemical staining for both markers compared to pretreatment staining. p53 and Ki-67 markers are associated with cell proliferation in children with EoE but do not represent a premalignant (dysplastic) condition of the esophagus.

Polyethylene glycol 3350 based colon cleaning protocol: 2 d vs 4 d head to head comparison.

AIM: To compare between 2 and 4 d colon cleansing protocols. METHODS: Children who were scheduled for colonoscopy procedure (2010-2012) for various medical reasons, were recruited from the pediatric gastroenterology clinic at Marshall University School of Medicine, Huntington, WV. Exclusion criteria were patients who were allergic to the medication used in the protocols [polyethylene glycol (PEG) 3350, Bisacodyl], or children with metabolic or renal diseases. Two PEG 3350 protocols for 4 d (A) and 2 d (B) were prescribed as previously described. A questionnaire describing the volume of PEG consumed, clinical data, and side effects were recorded. Colon preparation was graded by two observers according to previously described method. MAIN OUTCOME MEASUREMENTS: Rate of adequate colon preparation. RESULTS: A total of 78 patients were considered for final calculation (group A: 40, group B: 38). Age and stool consistency at the last day was comparable in both groups, but the number of stools/day was significantly higher in group B (P = 0.001). Adequate colon preparation was reached in 57.5% (A) and 73.6% (B), respectively (P = 0.206). Side effects were minimal and comparable in both groups. There was no difference in children's age, stool characteristics, or side effects between the children with adequate or inadequate colon preparation. Correlation and agreement between observers was excellent (Pearson correlation = 0.972, kappa = 1.0). CONCLUSION: No difference between protocols was observed, but the 2 d protocol was superior for its shorter time. Direct comparison between different colon cleansing protocols is crucial in order to establish the "gold standard" protocol for children.