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Nicolas Kalach, Nathalie Kapel,
Anne-Judith Waligora-Dupriet,
Marie-Christine Castelain,
Marie Odile Cousin,
Christine Sauvage,
Fatimata Ba,
Ioannis Nicolis,
Florence Campeotto,
Marie José Butel,
Christophe Dupont
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ABSTRACT: Abstract Background: Food allergy is a common problem in France involving 4%-6% of toddlers. As opposed to IgE-mediated cow's milk allergy (CMA), delayed-onset CMA, mostly, non-IgE-mediated, remains difficult to diagnose in toddlers. Our study assessed the diagnostic performances of intestinal permeability and of fecal markers, in comparison with the standard allergic work-up in children referred for CMA diagnosis. Methods: Twenty-five consecutive children, mean age (standard deviation) 6.3 months (4.8) with digestive and/or extra-digestive manifestations suggesting CMA, were prospectively studied based on a standardized allergic work-up (specific cow's protein IgE and IgG, skin prick test, atopy patch test and oral open cow's milk challenge) and digestive work-up including fecal microbiota analysis, intestinal permeability determination (urinary lactitol/mannitol ratio) and fecal markers measurement, i.e., α1-antitrypsin, tumor necrosis factor-α, calprotectin, β-defensin2, secretory IgA and eosinophil-derived neurotoxin (EDN). Receiver operating characteristic (ROC) curves were calculated for all markers in order to define cut-off levels. Results: The cow's milk challenge was positive in 11 children and negative in 14. The global test performances, i.e., the number of true positive+negative cases/the total number of cases, were 76% for intestinal permeability; 72% for fecal EDN; contrasting with atopy patch test, 68%; IgE, 60%; skin prick test, 55% and IgG, 52%. Conclusions: In this routine diagnosis allergy work-up for CMA in toddlers, the best efficacy was seen for intestinal permeability compared to IgE, IgG, skin prick test and atopy patch test. Moreover, fecal EDN in a single spot sample displayed a similar performance.
Clinical Chemistry and Laboratory Medicine 05/2012; · 2.15 Impact Factor
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Jean-Christophe Rozé,
Sébastien Barbarot,
Marie-José Butel, Nathalie Kapel,
Anne-Judith Waligora-Dupriet,
Inès De Montgolfier,
Magali Leblanc,
Nathalie Godon,
Pascale Soulaines,
Dominique Darmaun,
Montserrat Rivero,
Christophe Dupont
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ABSTRACT: The aim of the present study was to evaluate the safety, tolerance and preventive effect on atopic dermatitis of an experimental α-lactalbumin-enriched and symbiotic-supplemented infant formula. A total of ninety-seven non-breastfed term neonates were enrolled into a double-blind, multicentre, randomised controlled trial in which they received experimental (n 48) or standard formula (n 49) for 6 months. The primary outcome was weight at 6 months of age. Secondary outcomes were gastrointestinal tolerance and manifestation of atopic dermatitis. Faecal secretory IgA (SIgA) concentration and microbiota composition of forty-three infants were analysed at 1 and 6 months. Growth was similar in both groups. At 1 month, compared to those in the control group, infants in the experimental group exhibited less crying or agitation, and more quiet behaviour (P=0·03). At 6 months, atopic dermatitis was less frequently observed in the experimental group (P<0·05). Decrease of faecal SIgA concentration between 1 and 6 months was mainly observed in the control group. This decrease was significantly associated with atopic dermatitis (P<0·014) and negatively correlated to the level of colonisation by bifidobacteria (P<0·005). In conclusion, compared to the control formula, the experimental formula guaranteed a similar growth, was better tolerated at 1 month and had a protective effect against the development of atopic dermatitis.
The British journal of nutrition 11/2011; 107(11):1616-22. · 3.45 Impact Factor
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Florence Campeotto,
Antonia Suau, Nathalie Kapel,
Fabien Magne,
Vivian Viallon,
Laurent Ferraris,
Anne-Judith Waligora-Dupriet,
Pascale Soulaines,
Bernard Leroux,
Nicolas Kalach,
Christophe Dupont,
Marie-José Butel
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ABSTRACT: Intestinal bacterial colonisation in pre-term infants is delayed compared with full-term infants, leading to an increased risk of gastrointestinal disease. Modulation of colonisation through dietary supplementation with probiotics or prebiotics could decrease such a risk. The present study evaluated clinical tolerance, the effects on gut microbiota, and inflammatory and immunological mucosal responses to an infant formula adapted for pre-term infants that included in its manufacturing process a fermentation step with two probiotic strains, Bifidobacterium breve C50 and Streptococcus thermophilus 065, inactivated by heat at the end of the process. A total of fifty-eight infants (gestational age: 30–35 weeks), fed either the fermented pre-term formula or a standard pre-term formula, were followed up during their hospital stay. Clinical tolerance, faecal microbiota using a culture and a culture-independent method (temporal temperature gel electrophoresis), faecal calprotectin and secretory IgA were analysed weekly. No difference was observed regarding anthropometric data and digestive tolerance, except for abdominal distension, the incidence of which was lower in infants fed the fermented formula for 2 weeks. Bacterial colonisation was not modified by the type of feeding, particularly for bifidobacteria. Faecal calprotectin was significantly lower in infants fed the fermented formula for 2 weeks, and secretory IgA increased with both mother's milk and the fermented formula. The fermented formula was well tolerated and did not significantly modulate the bacterial colonisation but had benefits on inflammatory and immune markers, which might be related to some features of gastrointestinal tolerance.
The British journal of nutrition 06/2011; 105(12):1843 - 1851. · 3.45 Impact Factor
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ABSTRACT: The aim of this review was to examine the characteristics of the faecal calprotectin assay in neonates and the evidence for its use as a noninvasive marker of intestinal illnesses during the neonatal period.
Bibliographic searches were performed in the MEDLINE electronic database up to February 2010 looking for the following words (all fields): "infants" or "neonates" and "calprotectin." Twenty studies, in which 1180 neonates were enrolled, were selected.
During the neonatal period, calprotectin levels are characterized by significantly higher values in both healthy full-term and preterm infants during their first year of life compared with reference values established for children and adults. No difference was observed according to gestational age or birth weight, whereas a higher faecal calprotectin level was detected during intestinal distress in neonates with either inflammatory or patent digestive alterations. Despite high interindividual variations, cutoff levels are proposed to identify infants with a high risk of intestinal illnesses.
Compared with adults and children, healthy full-term and preterm neonates have high calprotectin levels. The measurement of calprotectin levels in faeces can be a promising noninvasive clinical screening test for intestinal distress in neonates.
Journal of pediatric gastroenterology and nutrition 11/2010; 51(5):542-7. · 2.18 Impact Factor
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ABSTRACT: The beneficial effects of Bifidobacterium are partly due to its immunostimulatory properties. These immunostimulatory properties may be linked to the presence of unmethylated CpG motifs specific to bacterial DNA, which may induce a TH1 response by activating Toll-like receptors (TLR). Using in silico analyses, PCR amplification, and dot blotting, we characterized the CpG content of various bifidobacterial strains and evaluated the immunostimulatory properties and genomic heterogeneity of these motifs in the genus. Our in silico study, based on entire genome sequences from five bifidobacterial strains, showed that Bifidobacterium genomes contain numerous CpG motifs, including 5'-purine-purine-CG-pyrimidine-pyrimidine-3' and 5'-purine-TCG-pyrimidine-pyrimidine-3' motifs, and biologically active sequences previously identified in lactic acid bacteria. We identified four CpG-rich sequences with Bifidobacterium longum NCC2705. Two sequences with a percent G+C of about 68% included 14 and 16 CpG motifs. Two sequences with a percent G+C of about 60% included 16 and 6 CpG motifs. These sequences induce the production of monocyte chemoattractant protein 1 (MCP-1) and tumor necrosis factor alpha (TNF-alpha) through a pattern of TLR9 stimulation on RAW 264.7 macrophages. No link could be established between their immunostimulatory properties, the number of CpG motifs, and percent G+C. We investigated inter- and intraspecies heterogeneity in 71 strains of various origins. These sequences were highly conserved in the genus. No link was found between the presence of the CpG-rich sequence and the origin of the strains (healthy, allergic, or preterm infants). The high frequency of CpG motifs in the DNA of Bifidobacterium may play an important role in the immunostimulatory properties of commensal or probiotic bifidobacterial strains.
Applied and environmental microbiology 03/2010; 76(9):2846-55. · 3.69 Impact Factor
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Florence Campeotto,
Mariella Baldassarre,
Marie-José Butel,
Vivian Viallon,
Flore Nganzali,
Pascale Soulaines,
Nicolas Kalach,
Alexandre Lapillonne,
Nicola Laforgia,
Guy Moriette,
Christophe Dupont, Nathalie Kapel
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ABSTRACT: This study aimed to determine cutoff levels for fecal calprotectin as a marker of intestinal distress in preterm neonates. A total of 126 infants born at a median gestational age of 33 weeks (range 25.7-35 weeks) were enrolled. Samples (n = 312) were collected weekly from the end of the first week of life until the end of the first month and if any gastrointestinal event occurred. Receiver operating characteristic curves analysis gave cutoff values of 363 microg/g (sensitivity 0.65, specificity 0.82) and 636 microg/g (sensitivity 0.72, specificity 0.95) for the development of mild or severe enteropathy.
Journal of pediatric gastroenterology and nutrition 05/2009; 48(4):507-10. · 2.18 Impact Factor
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ABSTRACT: Defensins, endogenous antibiotic peptides, are part of the intestinal epithelial barrier. In this pilot study we analyzed the possibility of measuring fecal beta-defensin-2 (HBD2) in comparing inflammatory and noninflammatory conditions. In samples from healthy control individuals, low levels of HBD2 were detectable, which markedly rose under inflammatory conditions (P = 0.0002 vs normal control individuals), the highest levels being observed in patients with ulcerative colitis (median 356 ng/g, range 40-527). Despite frank inflammation, Crohn disease patients with colitis had significantly lower, albeit enhanced, HBD2 levels than did ulcerative colitis patients. These data confirm the possibility of quantifying HBD2 in feces and indicate that colitis in Crohn disease and colitis in ulcerative colitis differ from each other with respect to their ability to secrete HBD2.
Journal of pediatric gastroenterology and nutrition 02/2009; 48(1):117-20. · 2.18 Impact Factor
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ABSTRACT: Cryptosporidium parvum is a protozoan parasite that causes intestinal malabsorptive syndrome and malnutrition. Considering the importance of di-tripeptide absorption for nutritional status, we previously investigated the regulation of PepT1 transporter in the suckling rat model of acute cryptosporidiosis and showed that PepT1 protein expression and activity were not modified in the parasitized intestine. Here we used confocal microscopy performed on intestinal villi to determine the subcellular localization of PepT1 together with f-actin and parasites. For this purpose, confocal microscopy using vibratome thick sections was developed on the distal small intestine, the preferential site of parasite implantation. Results showed major heterogeneity of apical PepT1 expression among enterocytes, which did not correlate with actin staining or parasite implantation. These results underscore the importance of considering the effect of C. parvum at the cellular scale and not only in the entire epithelium.
Parasitology Research 12/2008; 104(5):985-91. · 2.15 Impact Factor
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Acta Paediatrica 11/2007; 96(10):1531-3. · 2.07 Impact Factor
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ABSTRACT: Cryptosporidium parvum is a parasitic protozoa increasingly appreciated as a cause of intestinal malabsorptive syndrome leading to malnutrition and/or growth failure. Because a major mechanism for apical peptide absorption by small intestine is via the proton-coupled transporter PepT1, we investigated the expression and functionality of this transporter in our model of acute cryptosporidiosis. Four-day-old Sprague-Dawley rats were inoculated by gavage with 5 x 10(5) oocysts of C. parvum and killed at Day 12 (peak of the infection) or Day 21 (spontaneous clearance of the parasite). PepT1 expression and functionality were quantified in the distal small intestine, preferential site of C. parvum implantation, and in the proximal small intestine, free of parasite, using Western blot and Ussing chambers, respectively. No difference in total PepT1 protein expression or in glycyl-sarcosine fluxes was observed in C. parvum-infected rats compared with controls either on Day 12 or on Day 21, both in the proximal and in the distal small intestine. However, a significant decrease of apical membrane protein expression of PepT1 was observed in C. parvum-infected enterocytes compared with controls. This maintained dipeptide transport observed despite villous atrophy and decreased expression of the protein at the brush-border membrane strongly suggest a transient upregulation of PepT1 activity, probably related to gamma-interferon regulation.
Experimental Biology and Medicine 04/2007; 232(3):454-60. · 2.64 Impact Factor
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ABSTRACT: To date, there is no efficient treatment for cryptosporidiosis and parasite eradication relies on innate and acquired immunity. In this study, we investigated the effect of administration of probiotic bacteria on the development and progression of the experimental infection in suckling rats. Rats were fed daily with 2.10(7) CFU of Lactobacillus casei-containing mixture, starting 2 days before the infection until the spontaneous clearance of the parasite. Effects on weight gain, parasite burden, mucosal histology and production of mucosal cytokines (IFNgamma, IL10 and TNFalpha) were studied. Although a trend to a more rapid clearance of parasites was noted in rats treated with probiotics, no significant effect of probiotics administration was observed in terms of weight gain, parasite burden, mucosal damage, or kinetics of mucosal cytokines during the course of infection. Overall, our results showed that the daily administration of L. casei-containing mixtures was unable to eradicate the parasite in our model.
Parasitology Research 11/2006; 99(5):522-7. · 2.15 Impact Factor
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ABSTRACT: This study aimed to explore the metabolic consequences of cryptosporidiosis in an acute experimental model both at the peak of infection and after parasite clearance. Four-day-old suckling rats were infected with 10(6) oocysts of Cryptosporidium parvum. At the peak of infection (day 8 PI), C. parvum resulted in a dramatic reduction both in nutrient intake (-50%) and body weight (16.3+/-5.2 vs 27.3+/-1.0 g, P<0.01) with a decrease in both lean body mass and adipose tissue. Muscular fractional and absolute synthesis rate were reduced (-15 and -55%, respectively). After parasite clearance (day 17 PI), body weight remained reduced in formerly infected animals (37.8+/-8.0 vs 47.8+/-4.2 g, P<0.01) whereas nutrient intake normalized and fractional synthesis rate slightly increased (+22%) compared to controls. Overall, our results show that the impact and consequences of cryptosporidiosis are far greater than generally appreciated, leading to major malnutrition in suckling rats.
Parasitology Research 07/2005; 96(5):326-30. · 2.15 Impact Factor
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ABSTRACT: Controversy surrounds a 3-week treatment with a high-dose (0.14 mg. kg(-1). day(-1)) growth hormone (GH), glutamine, and high carbohydrate diet in home parenteral nutrition (HPN)-dependent patients with short-bowel syndrome (SBS). This study assessed treatment with low-dose GH in these patients.
Twelve adult HPN-dependent (duration, 7 +/- 1 years; mean +/- SEM) patients with SBS (small-bowel remnant length, 48 +/- 11 cm) who were on an unrestricted hyperphagic diet were randomized in a double-blind, placebo-controlled, crossover study. Patients received daily low-dose GH (0.05 mg. kg(-1). day(-1)) and placebo for two 3-week periods separated by a 1-week washout period. Net intestinal absorption of macronutrients was assessed using a duplicate diet; nutritional assessment and blood tests were performed. Data from each group were compared using Wilcoxon rank sum test.
Treatment with GH increased intestinal absorption of energy (15% +/- 5%, P < 0.002), nitrogen (14% +/- 6%, P < 0.04), carbohydrates (10% +/- 4%, P < 0.04), and fat (12% +/- 8%, NS). The increased food absorption represented 37% +/- 16% of total parenteral energy delivery. Body weight (P < 0.003), lean body mass (P < 0.006), D-xylose absorption (P < 0.02), insulin-like growth factor 1 (P < 0.002), and insulin-like growth factor binding protein 3 (P < 0.002) increased, whereas uptake of GH binding protein decreased (P < 0.01), without any major adverse effect.
Three weeks of low-dose GH significantly improved intestinal absorption in HPN-dependent SBS patients who were on a hyperphagic western diet.
Gastroenterology 02/2003; 124(2):293-302. · 11.68 Impact Factor
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ABSTRACT: The ontogenetic development of PepT1, NBAT and EAAC1 along the vertical and horizontal axes of the rat small intestine was evaluated using semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry. The proximodistal profiles of mRNA levels showed that PepT1 was evenly distributed, whereas NBAT had greater expression in the proximal part, and EAAC1 in the distal part. These regionalizations were the same from postnatal days 4 to 50. PepT1 and NBAT proteins were detected in the microvilli of enterocytes along the length of the villi. NBAT was also found in the cytoplasm. Surprisingly, EAAC1 was located exclusively in the microvilli of enterocytes in the crypt and the bases of the villi. These protein expression patterns were similar in all parts of the small intestine (proximal, median and distal), at all ages. We conclude that the expression of PepT1, NBAT or EAAC1 are differently regulated according to both the horizontal and vertical axes.
Journal of Nutrition 06/2002; 132(5):1009-11. · 3.92 Impact Factor
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ABSTRACT: This prospective study aimed to establish the effect of recombinant human growth hormone (rhGH) on intestinal function in children with short bowel syndrome (SBS). Eight children with neonatal SBS were included. All were dependent on parenteral nutrition (PN) for >3 years (range, 3.8-11.6 years), with PN providing >50% of recommended dietary allowance for age (range, 50%-65%). The subjects received rhGH (Humatrope) 0.13 mg/kg/d subcutaneously over a 12-week period. The follow-up was continued over a 12-month period after rhGH discontinuation. Clinical and biological assessments were performed at baseline, at the end of the treatment period, and 12 months after the end of treatment. No side effects related to rhGH were observed. PN requirements were decreased in all children during the course of rhGH treatment. Between baseline and the end of treatment, significant increases were observed in concentrations (mean ± standard deviation) of serum insulin-like growth factor 1 (103.1 ± 49.9 µg/L vs 153.5 ± 82.2 µg/L; P < .01), serum insulin-like growth factor-binding protein 3 (1.7 ± 0.6 mg/L vs 2.5 ± 0.9 mg/L; P < .001), and plasma citrulline (16.5 ± 14.8 µmol/L vs 25.2 ± 18.3 µmol/L; P < .05). A median 54% increase in enteral intake (range, 10%-244%) was observed (P < .001) and net energy balance improved significantly (P < .002). It was necessary for 6 children to be maintained on PN or restarted after discontinuation of rhGH treatment, and they remained on PN until the end of the follow-up period. A 12-week high-dose rhGH treatment allowed patients to decrease PN, but only 2 patients could be definitively weaned from PN. Indications and cost-effectiveness of rhGH treatment for SBS pediatric patients need further evaluation.
Journal of Parenteral and Enteral Nutrition 34(5):513-20. · 3.29 Impact Factor
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ABSTRACT: Capet, C., Kapel, N., Huneau, J. F., Magne, D., Laikuen, R., Tricottet, V., Benhamou, Y., Tomé, D., and Gobert, J. G. 1999.Cryptosporidium parvuminfection in suckling rats: Impairment of mucosal permeability and Na+-glucose cotransport.Experimental Parasitology91,119–125. Na+–glucose transport and transepithelial permeability were investigated during symptomatic acute cryptosporidiosis in newborn rats. The infection resulted in a significant (P< 0.01) decrease in the ileal short-circuit current and a nonsignificant fall in the transepithelial potential difference and conductance. In glucose-stimulated conditions, the rise in ileal short-circuit current and transepithelial permeability were significantly lower inCryptosporidium parvum-infected rats than in controls (ΔIsc = 3.24 ± 1.21 μA·cm−2vs ΔIsc = 5.09 ± 2.23 μA·cm−2in infected and control animals, respectively;P< 0.001; ΔPD = −0.35 ± 0.13 mV vs ΔPD = −0.44 ± 0.14 mV for infected and control animals, respectively;P< 0.01). Electrical parameters were not affected by addition of the cyclooxygenase inhibitor indomethacin in eitherCryptosporidium-infected newborn rats or controls. Horseradish peroxidase and mannitol flux studies demonstrated a significant decrease (P< 0.05) in transepithelial molecular permeability in infected enterocyte rats, HRP flux = 380, range 68–5570 ng·cm−2, and mannitol flux = 1.06, range, 0.34–1.44% ·cm−2·min−1, compared with controls rats, HRP flux = 4446 range, 1121–124,363 ng·cm−2, and mannitol flux = 1.99, range, 0.57–5.09%·cm−2·min−1;P< 0.05. These effects could originate fromC. parvum-induced alteration of intracellular trafficking of pinocytosis vesicles and therefore account for the decrease in permeability to solute and macromolecules, together with impaired transcellular nutrient transport, in suckling rats.
Experimental Parasitology 91(2):119-125. · 2.12 Impact Factor
