Influence of urine volume on the assessment of intestinal permeability in affected children by multiple sugar probes.

BACKGROUND: In this study we have looked at the reliability of a multi-sugar test in a pediatric patient population and its accuracy at small urine volumes to evaluate intestinal permeability. METHODS: Out of 117 subjects enrolled, 31 were healthy and 86 were sick. A solution containing lactulose, rhamnose, sucrose, and sucralose was administered to subjects who were on fasting; the urine excreted during 5 h was collected and measured. Samples were analyzed by gas chromatography-tandem mass spectrometry and results were expressed as percentage of sugar recoveries and lactulose/rhamnose (L/R) ratio. RESULTS: The analyses showed a clear effect of low urinary volumes (≤240 mL) particularly affecting rhamnose excretion in healthy subjects and sucrose and sucralose recovery in diseased children. Despite the low rhamnose recovery, as lactulose is not similarly affected, the diagnostic reliability of L/R ratio is well preserved at low diuresis conditions. However, this ratio can be useful to discriminate acute conditions vs. clinical remissions only at high urine volumes. Data also suggest potential diagnostic applicability of sucrose and sucralose in children at high urine volumes. CONCLUSIONS: In conclusion, the multi-sugar test has a good predictivity in pediatric subjects but results must be carefully interpreted in the face of reduced diuresis.

Rapid anti-transglutaminase assay and patient interview for monitoring dietary compliance in celiac disease.

OBJECTIVE: The anti-transglutaminase antibodies (anti-tTG) play an important role in monitoring the celiacs' gluten-free diet (GFD). MATERIAL AND METHODS: The authors propose to use the rapid IgA anti-tTG assay based on a whole blood drop to evaluate the compliance to GFD at the clinical ambulatory setting. The rapid test results were compared with those of the conventional ELISA assay and with dietary compliance reported by patients' interview. CONCLUSIONS: The authors showed that anti-tTG rapid test is reliable and easy to perform in the ambulatory setting to evaluate dietary compliance. Moreover, they proved that celiacs' interview is more sensitive than serology in identifying patients who transgress.

Search for atoxic cereals: a single blind, cross-over study on the safety of a single dose of Triticum monococcum, in patients with celiac disease.

BACKGROUND: Cereals of baking quality with absent or reduced toxicity are actively sought as alternative therapy to a gluten-free diet (GFD) for patients with coeliac disease (CD). Triticum monococcum, an ancient wheat, is a potential candidate having no toxicity in in-vitro and ex-vivo studies. The aim of our study was to investigate on the safety of administration of a single dose of gluten of Tm in patients with CD on GFD. METHODS: We performed a single blind, cross-over study involving 12 CD patients who had been on a GFD for at least 12 months, challenged on day 0, 14 and 28 with a single fixed dose of 2.5 grams of the following (random order): Tm, rice (as reference atoxic protein) and Amygluten (as reference toxic protein) dispersed in a gluten-free pudding. The primary end-point of the study was the change in intestinal permeability, as assessed by changes in the urinary lactulose/rhamnose ratio (L/R ratio) measured by High Pressure Liquid Chromatography. We also assessed the occurrence of adverse gastrointestinal events, graded for intensity and duration according to the WHO scale. Variables were expressed as mean ± SD; paired t-test and χ² test were used as appropriate. RESULTS: The urinary L/R ratio did not change significantly upon challenge with the 3 cereals, and was 0.055 ± 0.026 for Tm Vs 0.058 ± 0.035 for rice (p = 0.6736) and Vs 0.063 ± 0.054 with Amygluten (p = 0.6071). Adverse gastrointestinal events were 8 for Tm, Vs 11 for rice (p = 0.6321) and Vs 31 for Amygluten p = 0.0016), and, in all cases events were graded as "mild" or "moderate" with TM and rice, and as "severe" or "disabling" in 4 cases during Amygluten. CONCLUSIONS: No definite conclusion can be drawn on the safety of Tm, based on no change in urinary L/R because even Amygluten, a toxic wheat protein, did not cause a significant change in urinary L/R indicating low sensitivity of this methodology in studies on acute toxicity. Tm was, however, well tolerated by all patients providing the rationale for further investigation on the safety of this cereal for CD patients. TRIAL REGISTRATION: EudraCT-AIFA n2008-000697-20.

The active Zot domain (aa 288-293) increases ZO-1 and myosin 1C serine/threonine phosphorylation, alters interaction between ZO-1 and its binding partners, and induces tight junction disassembly through proteinase activated receptor 2 activation.

Vibrio cholerae-derived zonula occludins toxin (Zot) is a multifunctional protein that reversibly disassembles intestinal tight junctions (tjs). Zot structure-function analysis has mapped this activity to aa 288-293, named AT1002. AT1002 reduced transepithelial electrical resistance across rat small intestine, ex vivo, as did Zot and its processed mature form, ΔG. AT1002 increased in vivo permeability to sugar tracers, whereas scrambled control peptides did not. Binding and barrier assays in proteinase activated receptor (PAR)(2)-expressing and PAR(2)-null cells established AT1002 activity to be PAR(2) dependent. Coincident with the increased intestinal permeability, confocal microscopy of AT1002-exposed rat intestinal IEC6 cells revealed displacement of ZO-1 and occludin from intercellular boundaries. In coimmunoprecipitation assays, AT1002 decreased ZO-1-occludin and ZO-1-claudin 1 interactions coincident with PKCα-dependent ZO-1 serine/threonine phosphorylation. Further, AT1002 increased serine phosphorylation of myosin 1C and, at the same time, transiently diminished its association with ZO-1. The COOH-terminal domain of ZO-1 was required for its association with myosin 1C. These data indicate that the NH(2)-terminal portion of active Zot contains a PAR(2)-activating motif, FCIGRL, that increases PKCα-dependent ZO-1 and myosin 1C serine/threonine phosphorylation. These modifications provoke selective disengagement of ZO-1 from its binding partners, occludin, claudin 1, and myosin 1C, coincident with opening of tjs.

Prevalence of breastfeeding in a baby-friendly pediatric practice in Trieste, Italy: follow up to 36 months of age.

BACKGROUND: A breastfeeding-friendly physician's office that applies the 13 recommendations of the Academy of Breastfeeding Medicine can help increase the exclusivity and duration of breastfeeding. Having already published the results up to five months of age of this intervention in our pediatric practice, we now report on the follow up to 36 months. METHODS: A cohort of 252 newborn infants was enrolled with our pediatric office in Trieste, Italy, between 1 January 2016 and 31 December 2016. The office implemented baby-friendly pediatric practices and a biological nurturing approach to the support of breastfeeding. In addition to the services offered by two pediatricians, support was provided by a peer counselor. Data on breastfeeding were collected at periodic healthy child visits up to 36 months of age. The outcome of interest for this follow up was the rate of any breastfeeding, defined as the percentage of infants and children who had received breastmilk in the previous 24 h. RESULTS: The rates of any breastfeeding at discharge and at 1, 3 and 5 months (n = 252) were 95.2, 95.8, 89.3 and 86.5%, respectively. At 8, 12, 18, 24 and 36 months of age, the rates of breastfeeding were 70.6% (163/231), 59% (135/229), 35% (78/224), 24.6% (55/224) and 7.2% (16/224), respectively. CONCLUSIONS: The rates of any breastfeeding recorded in our pediatric practice up to age 36 months, are much higher than those reported elsewhere in high income countries and are likely to be associated with our baby-friendly and biological nurturing approach.

Fasting increases tobramycin oral absorption in mice.

The pharmacokinetics of the aminoglycoside tobramycin was evaluated after oral administration to fed or fasting (15 h) mice. As expected, under normal feeding conditions, oral absorption was negligible; however, fasting induced a dramatic increase in tobramycin bioavailability. The dual-sugar test with lactulose and l-rhamnose confirmed increased small bowel permeability via the paracellular route in fasting animals. When experiments aimed at increasing the oral bioavailability of hydrophilic compounds are performed, timing of fasting should be extremely accurate.

Prevalence of breastfeeding in a baby-friendly pediatric practice: an experience in Trieste, Italy.

Background: In a pediatric practice in Italy, actions were undertaken to apply the recommendations for a breastfeeding-friendly physician's office and to promote the adoption of a semi-reclined or laid-back maternal position in breastfeeding. The aim of this study is to evaluate the effect of the actions implemented, in terms of prevalence of exclusive breastfeeding. Methods: A historical cohort study was carried out using administrative data routinely collected. All women who gave birth in 2016 and registered their newborns with the pediatric practice were included, only mothers of preterm newborns < 30 weeks gestational age were excluded. The main actions undertaken were: employment of a breastfeeding peer supporter; ensuring unlimited daily access in case of breastfeeding difficulties; provision of individual support to breastfeeding mothers in a dedicated room and advice on the laid-back position; scheduling of weekly meetings of small groups for breastfeeding support. Each infant was followed up for five months. The main study outcomes were duration of exclusive breastfeeding (only breast milk and no other liquids or solids, except for drops of syrups with nutritional supplements or medicines) and prevalence at five months. Results: A total of 265 newborn infants with a gestational age greater than 30 weeks were registered with the pediatric practice during the study period, about 18% of all infants born in Trieste in that period. Complete data were available for 252 of these (95.1%). The rate of exclusive breastfeeding at five months of age was higher than the one reported for the whole infant population of Trieste and of the Friuli Venezia Giulia Region (62.3% vs. 42.9% vs. 30.3%) in the same period. Conclusions: The implementation of breastfeeding-friendly pediatric practice and the application of laid-back breastfeeding may improve the rate and duration of exclusive breastfeeding.

Anti-α-enolase Antibodies in Serum from Pediatric Patients Affected by Inflammatory Diseases: Diagnostic and Pathogenetic Insights.

Human glycolytic enzyme α-enolase was associated with human diseases and with inflammation. An ELISA test was developed to measure anti-α-enolase AAE IgG and AAE IgA in the serum from patients affected by inflammatory diseases with the purpose to evaluate it as a novel diagnostic marker. 80 healthy blood donors and 194 paediatric patients affected by Juvenile idiopathic arthritis (JIA), celiac disease (CD), Crohn's Disease (CrD), hereditary periodic fever (HPF), and PFAPA syndrome were included in the study. HPF patients showed high levels of AAE antibodies, whereas JIA, CD, and CrD presented only partial results. Benign fevers such as PFAPA were almost negative for AAE Abs. These findings suggested that the genetic dysfunction of inflammasome associated with HPF could lead to the formation of AAE Abs that could be used for an early and easy diagnosis.

Speeding up coeliac disease diagnosis in cardiological settings.

INTRODUCTION: High prevalence of coeliac disease (CD) has been reported among patients with idiopathic dilated cardiomyopathy (DCM). We evaluated the feasibility and diagnostic accuracy of screening for CD by rapid test of anti-transglutaminase antibodies in the cardiology outpatients' clinic. MATERIAL AND METHODS: We screened the blood samples of 104 patients with DCM, 44 of their first-degree relatives, 63 diseased controls and 101 healthy controls for the presence of anti-transglutaminase antibodies in a drop of whole blood using a rapid assay. This test was compared to the enzyme-linked immunosorbent assay and the anti-endomysium antibody test. RESULTS: Our rapid test was positive in three (2.9%) DCM patients, in one (2%) relative and in one (1%) healthy control. These subjects were positive at both control assays. Two DCM patients had iron-deficient anaemia. The healthy relative was asymptomatic, while the healthy control experienced extreme asthenia. The relative refused intestinal biopsy, while the others showed histological evidence of CD. During the gluten-free diet, the patient with the worst left ventricular ejection fraction (LVEF) underwent heart transplant, and LVEF values improved in the other two. Anaemia and tiredness resolved in all patients. CONCLUSION: Early detection of CD in a cardiological setting allows prompt treatment with a gluten-free diet of gluten-dependent complaints with potential benefits for the course of DCM.