Pdx1 and controlled culture conditions induced differentiation of human amniotic fluid-derived stem cells to insulin-producing clusters.

This study investigated the differentiation of human amniotic fluid-derived stem cells (hAFSCs) into insulin-producing clusters in vitro. Adenovirally-delivered mouse Pdx1 (Ad-Pdx1) induced human Pdx1 expression in hAFSCs and enhanced the coordinated expression of downstream β-cell markers. When Ad-Pdx1-transduced hAFSCs were sequentially treated with activin A, bFGF and nicotinamide and the culture plate surface coated with poly-l-ornithine, the expression of islet-associated human mRNAs for Pdx1, Pax6, Ngn3 and insulin was increased.

Substrate elasticity controls cell proliferation, surface marker expression and motile phenotype in amniotic fluid-derived stem cells.

The physical cues presented to stem cells by the substrate on or in which they exist have been shown to play a crucial role in regulation of their behavior. Until recently, most research has focused on the effects of substrate elasticity on differentiation capability rather than maintenance of long-term proliferation and plasticity. The main goal of the present study is to study the interaction of amniotic fluid-derived stem (AFS) cells with growth substrata of different elasticity, which may extend their application potentials.

Outcome following aminosalicylate therapy in children newly diagnosed as having ulcerative colitis.

Objectives: Despite a paucity of published supporting data, 5-aminosalicylate (5-ASA) use in pediatric ulcerative colitis (UC) is common. The present study describes the use and outcome of a large multicenter inception cohort of children with UC treated with 5-ASA.

Methods: Data were obtained from the Pediatric Inflammatory Bowel Disease Collaborative Research Group Registry, a prospective North American observational study of children newly diagnosed as having inflammatory bowel disease ages 16 years or younger.

Budesonide use in pediatric Crohn disease.

Background And Aim: Budesonide (BUD) is being used in pediatric Crohn disease (CD) because it is believed to have the potential to reduce corticosteroid-related toxicity; however, few data are available describing its use. The aim of the present study was to describe BUD use in an inception cohort of pediatric patients with CD.

Methods: Data were derived from the prospective Pediatric IBD Collaborative Research Group Registry established in 2002 in North America.

Probiotics in inflammatory bowel diseases and associated conditions.

A complex set of interactions between the human genes encoding innate protective functions and immune defenses and the environment of the intestinal mucosa with its microbiota is currently considered key to the pathogenesis of the chronic inflammatory bowel diseases (IBD). Probiotics offer a method to potentially alter the intestinal microbiome exogenously or may provide an option to deliver microbial metabolic products to alter the chronicity of intestinal mucosal inflammation characterizing IBD. At present, there is little evidence for the benefit of currently used probiotic microbes in Crohn's disease or associated conditions affecting extra-intestinal organs.

Alterations in the gut microbiome of children with severe ulcerative colitis.

Background: Although the role of microbes in disease pathogenesis is well established, data describing the variability of the vast microbiome in children diagnosed with ulcerative colitis (UC) are lacking. This study characterizes the gut microbiome in hospitalized children with severe UC and determines the relationship between microbiota and response to steroid therapy.

Methods: Fecal samples were collected from 26 healthy controls and 27 children hospitalized with severe UC as part of a prospective multicenter study.

Spontaneous transformation of murine epithelial cells requires the early acquisition of specific chromosomal aneuploidies and genomic imbalances.

Human carcinomas are defined by recurrent chromosomal aneuploidies, which result in a tissue-specific distribution of genomic imbalances. In order to develop models for these genome mutations and to determine their role in tumorigenesis, we generated 45 spontaneously transformed murine cell lines from normal epithelial cells derived from bladder, cervix, colon, kidney, lung, and mammary gland. Phenotypic changes, chromosomal aberrations, centrosome number, and telomerase activity were assayed in control uncultured cells and in three subsequent stages of transformation.

Associations between variants in the ABCB1 (MDR1) gene and corticosteroid dependence in children with Crohn's disease.

Background: Corticosteroids (CS) effectively induce remission in patients with moderate to severe Crohn's disease (CD). However, CS dependence in children is a significant clinical problem associated with numerous side effects. Identification of molecular markers of CS dependence is of paramount importance.

C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) or both? A systematic evaluation in pediatric ulcerative colitis.

Background: There has not been an extensive comparison of CRP and ESR in ulcerative colitis (UC), and thus, we aimed to explore their utility in UC.

Methods: Four previously enrolled cohorts of 451 children with UC were utilized, all including laboratory, clinical and endoscopic data. A longitudinal analysis was performed on prospectively collected data of 75 children.

Isolation of mesenchymal stem cells from the mandibular marrow aspirates.

Objective: The aim of this study was to determine if mesenchymal stem cells (MSCs) could be obtained from mandible marrow aspirates.

Study Design: In 5 patients, 10 mL marrow aspirates were obtained from both the mandible and the iliac crest. Second passage MSCs were characterized by fluorescence-activated cell sorting (FACS) analysis using MSC-specific cell surface marker, and fifth-passage cells were differentiated into 3 mesenchymal tissues in vitro.

Maternal separation and gastrointestinal transit time in neonate rats.

Gastrointestinal transit times (GItts) were compared in separate litters of 10- and 15-day-old Sprague Dawley rats using barium sulphate. By tracking the leading front of the bolus on radiographs, the gastrocaecal transit times in pups were estimated. To measure the total GItt, the duration from orogastric gavage until an observable defecation of barium sulphate was recorded.

Fecal osteoprotegerin may guide the introduction of second-line therapy in hospitalized children with ulcerative colitis.

Background: Osteoprotegerin (OPG) is increased in inflamed colonic mucosa and has a role in immune regulation and apoptosis resistance. Fecal OPG may be useful in predicting corticosteroid resistance in hospitalized children with severe ulcerative colitis (UC). We aimed to determine whether fecal OPG predicts the need for second-line therapies in children hospitalized for UC.

Short-course ondansetron for the prevention of methotrexate-induced nausea in children with Crohn disease.

Objectives: Methotrexate (MTX) is an effective treatment for Crohn disease (CD); however, its application may be limited by the occurrence of nausea. We assessed whether a short course of ondansetron minimized this adverse event.

Patients And Methods: A retrospective case-control study of patients with CD who received MTX at the Children's Hospital of Eastern Ontario between 2001 and 2009 was conducted.

Use of C-reactive protein in children with newly diagnosed inflammatory bowel disease.

C-reactive protein (CRP), a marker for inflammation, was evaluated with other routine blood tests in children with newly diagnosed inflammatory bowel disease. Evaluation of CRP level helped identify additional patients found to have inflammatory bowel disease at endoscopy, although a sizeable number of patients with mild ulcerative colitis had a normal CRP level.

Variation in the glucocorticoid receptor gene (NR3C1) may be associated with corticosteroid dependency and resistance in children with Crohn's disease.

Objectives: In pediatric onset of Crohn's disease (CD), corticosteroid dependency (approximately 40%) and resistance (approximately 10%) are significant clinical problems. Given the known effects of the glucocorticoid receptor (GR/NR3C1) gene in corticosteroid metabolism, we investigated whether variation in the gene was associated with corticosteroid response.

Methods: A retrospective cohort study was carried out including patients with CD diagnosed before 18 years and treated with a first course of corticosteroids in two Canadian tertiary pediatric gastroenterology clinics.

NELL1, NCF4, and FAM92B genes are not major susceptibility genes for Crohn's disease in Canadian children and young adults.

Background: Genome-wide association studies (GWAS) and replication studies have shown conflicting associations between the NELL1, NCF4, and FAM92B genes and susceptibility for Crohn's disease (CD). We sought to examine whether these genes were associated with CD in Canadian children and young adults.

Methods: A case-control study was carried out at three pediatric gastroenterology clinics across Canada.

Solid-food introduction in infants diagnosed as having a cow's-milk protein-induced enterocolitis.

Objectives: In a prior retrospective analysis of 32 infants diagnosed with cow's-milk protein-induced enterocolitis (CMPIE), gastrointestinal (GI) symptoms were observed with the introduction of milk-free infant cereal. The aim of this randomized controlled trial was to compare the incidence of new GI symptoms with the introduction of either milk-free infant rice cereal or carrots as the first complementary food at 6 months of age in infants previously diagnosed as having CMPIE.

Patients And Methods: Thirty-nine infants were enrolled in the study.

Lactobacillus plantarum 299v Prevents Caspase-Dependent Apoptosis In Vitro.

Selective microbes used as probiotics can enhance epithelial cell protection. We have previously shown that a Lactobacillus plantarum strain 299v (Lp299v) has the ability to induce mucin genes. In the current study, we utilized a cytokine model of inflammation in cell culture to study the modulation of apoptosis by this probiotic.

Mathematical weighting of the pediatric Crohn's disease activity index (PCDAI) and comparison with its other short versions.

Background: The Pediatric Crohn's Disease Activity Index (PCDAI) has become the standard outcome measure in pediatric Crohn's disease (CD) clinical research. Other versions have been proposed but without systematic evaluation. The aim was to assess validity and responsiveness of the abbreviated PCDAI (abbrPCDAI), short PCDAI (shPCDAI), and modified PCDAI (modPCDAI) as measures of disease activity and to compare these with a mathematically weighted version developed here (wPCDAI).

Safety assessment of probiotics for human use.

The safety of probiotics is tied to their intended use, which includes consideration of potential vulnerability of the consumer or patient, dose and duration of consumption, and both the manner and frequency of administration. Unique to probiotics is that they are alive when administered, and unlike other food or drug ingredients, possess the potential for infectivity or in situ toxin production. Since numerous types of microbes are used as probiotics, safety is also intricately tied to the nature of the specific microbe being used.

Outcome following thiopurine use in children with ulcerative colitis: a prospective multicenter registry study.

Objectives: Despite little supporting data, thiopurine use is common in pediatric ulcerative colitis (UC). Our aim was to determine outcome following thiopurine use in a multicenter inception cohort of children diagnosed with UC.

Methods: Data were obtained from a prospective observational study of newly diagnosed children <16 years of age.

Genes involved in the metabolism of poly-unsaturated fatty-acids (PUFA) and risk for Crohn's disease in children & young adults.

Background And Objectives: Epidemiological evidence for the role of polyunsaturated fatty-acids (PUFA) in Crohn's disease (CD) is unclear, although the key metabolite leucotriene B4 (LTB(4)) is closely linked to the inflammatory process. We hypothesized that inherited variation in key PUFA metabolic enzymes may modify susceptibility for CD.

Methods And Principal Results: A case-control design was implemented at three pediatric gastroenterology clinics in Canada.

Pulse probiotic administration induces repeated small intestinal Muc3 expression in rats.

Upon ingestion, probiotics may act to protect the host through a number of protective mechanisms including modulation of genes involved in intestinal innate mucosal defense such as epithelial cell-derived mucin glycoproteins and inhibitor of apoptosis proteins. To determine the specificity of effect and sustainability of response in vivo, Lactobacillus plantarum 299v (Lp299v), Lactobacillus rhamnosus R0011 (LrR0011), and Bifidobacterium bifidum R0071 (BbR0071) were added repeatedly or intermittently to the drinking water of Sprague-Dawley rats. After killing the rats via CO2 suffocation, Muc2, Muc3, neuronal apoptosis inhibitor protein (NAIP), human inhibitor of apoptosis protein 1/cellular inhibitor of apoptosis 2 (HIAP1/cIAP2), and human inhibitor of apoptosis protein 2/cellular inhibitor of apoptosis 1 (HIAP2/cIAP1) mRNA and protein levels were analyzed via RT-PCR and immunohistochemistry.

(14)C radiolabeling of proteins to monitor biodistribution of ingested proteins.

The economical preparation of microgram quantities of (14)C-labeled proteins by in vacuo methylation with methyl iodide is described. The (14)C radiolabeling was achieved by the covalent attachment of [(14)C]methyl groups onto amino and imidazole groups by reaction in vacuo with [(14)C]methyl iodide. The method was tested by investigating the biodistribution of (14)C in rats that were fed (14)C-labeled human soluble cluster of differentiation 14 (CD14) protein, a receptor for bacterial lipopolysaccharide.

Gene expression changes associated with resistance to intravenous corticosteroid therapy in children with severe ulcerative colitis.

Background And Aims: Microarray analysis of RNA expression allows gross examination of pathways operative in inflammation. We aimed to determine whether genes expressed in whole blood early following initiation of intravenous corticosteroid treatment can be associated with response.

Methods: From a prospectively accrued cohort of 128 pediatric patients hospitalized for intravenous corticosteroid treatment of severe UC, we selected for analysis 20 corticosteroid responsive (hospital discharge or PUCAI ≤45 by day 5) and 20 corticosteroid resistant patients (need for second line medical therapy or colectomy, or PUCAI >45 by day 5).