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Current studies indicate a link between the intake of exclusive enteral nutrition (EEN) and the induction of complex changes in the intestinal microbiota, as well as the clinical improvement of Crohn's disease (CD). The first aim of this study was to test the ability of various commensal bacterial strains (

Enteral nutrition (EN) formulas of polymeric type ordinarily have similar content of intact macronutrients but may vary in prebiotic saccharides and micronutrients. These components can play an important role in the intestinal microbiota modulation. The aim of this study was to investigate microbial changes of faecal samples after their in vitro anaerobic cultivation in four polymeric EN formulas using plate technique method, metabolite analysis, and microbiota profiling using 16S rRNA sequencing. Detected cultivable commensal groups (bifidobacteria, lactobacilli, Escherichia coli) in faecal samples of donors were able to grow in EN formulas. However, their counts varied depending on the individual donor and the type of EN formula. Similar trend was found in detected metabolites such as acetate, lactate, and butyrate. Also, taxonomic composition and diversity of original and cultivated faecal microbiota of one individual on different EN formula indicate a possible effect of the prebiotics and micronutrients to modulate gut microbiota.

Exclusive enteral nutrition (EEN) has been recommended as the first-line therapy in children with active Crohn disease (CD). The primary aim of our study was to determine whether it is possible to use the difference between basal fecal calprotectin (F-CPT) and the value at week 2 of EEN to predict clinical response at week 6. We prospectively collected stool samples for F-CPT analysis and clinical and laboratory parameters during EEN from 38 pediatric patients (28 boys, median age 12.8 years) with newly diagnosed active luminal CD. The difference between F-CPT concentrations before EEN and at week 2 did not predict clinical non-response at week 6 (OR 0.9996 95% CI 0.9989-1.0002, p = 0.18); however, it predicted patients who did not achieve clinical remission at week 6 (OR 0.9993, 95% CI 00.9985-0.9998, p = 0.006) with sensitivity of 58%, and specificity of 92% for cut-off of F-CPT increase by 486 μg/g.Conclusions: An early decrease in F-CPT levels in children with newly diagnosed active luminal CD did not predict clinical response at week 6 of EEN induction therapy, and clinical remission was predicted with low accuracy. Therefore, F-CPT cannot be used as a predictor to select the patients in whom EEN should be terminated. What is Known: • The fecal calprotectin (F-CPT) is an important marker of intestinal inflammation. • Approximately 25% of pediatric patients with Crohn disease (CD) do not achieve clinical remission, and there is still no sufficient predictor of response to exclusive enteral nutrition (EEN) treatment. What is New: • The difference between the F-CPT concentrations before EEN treatment and at week 2 did not predict clinical response to treatment at week 6, even if it predicted clinical remission, however, with low accuracy. F-CPT is not a suitable predictor to select the patients for discontinuing of EEN induction therapy.

Systemic lupus erythematosus (SLE) is known to imitate various diseases. We present a patient with failure to thrive and recurrent cystitis who presented abruptly with severe gastrointestinal manifestations requiring surgical treatment. Crohn’s disease of stomach was the most likely diagnosis, although no granuloma was found in tissue biopsies. Despite intensive immunosuppressive therapy, deep ulcers did not fully heal. Five years after disease manifestation, the patient developed lupus nephritis with autoimmune haemolytic anaemia and finally a diagnosis of SLE was made. Adequate management led to significant improvements in the patient’s clinical condition and laboratory results. In conclusion, diagnosis of SLE may be difficult mainly in cases with only GI and urinary symptoms.

Introduction : Biliary cysts with an abnormal pancreatobiliary junction are one of the most common pancreatobiliary malformations. The main symptom is cholestasis; endoscopic retrograde cholangiopancreatography (ERCP) plays a key role in diagnosis. Aim : Retrospective evaluation of ERCP performed to diagnose abnormalities of the pancreatobiliary junction. Material and methods : We retrospectively evaluated ERCP performed to diagnose abnormalities of the pancreatobiliary junction, mainly choledochal cysts, in 112 children between 1990 and 2011. Results : We performed 112 examinations of 50 children with abnormal pancreatobiliary junction and choledochal cysts (15 males and 35 females, average age: 5 years, range: 1 month – 15 years). Cysts were associated with a common channel in 37 (74%) cases, were not associated with a common channel in 9 (18%) cases, and in 3 (6%) cases the common channel lacked cysts. We performed endoscopic papilla sphincterotomy on 33 (66%) patients; endoscopic drainage was performed 62 times, including 17 patients without papilla sphincterotomy. In 15 (30%) cases, we only performed sphincterotomy. Extraction of lithiasis was done in 2 (4%) cases. Both ERCP and magnetic resonance cholangiopancreatography (MRCP) were performed on 13 patients. There was concordance of the choledochal cyst and of the type of cyst in both methods; however, common channels could not be observed by MRCP. There was 1 serious complication (perforation after sphincterotomy) and 11 (9%) mild adverse events. Conclusions : When performed at an expert centre, ECRP is a safe and reliable procedure for children with choledochal cysts and/or an abnormal pancreatobiliary junction. Endoscopic retrograde cholangiopancreatography remains a reasonable alternative because MRCP has a limited ability to diagnose the precise anomalies of the pancreatobiliary junction and has no therapeutic capabilities.

Background This study aimed to assess the declared benefits of the new test using antibodies against tissue transglutaminase in complex with gliadin representing a neo-epitope in the IgA and IgG class of immunoglobulins compared with currently used tissue transglutaminase antibodies in the IgA class of immunoglobulins among children. Methods In the cross-sectional study (P1 study, n = 406) and two small-size prospective observational studies (P2 study, n = 59 and P3 study, n = 12), serum samples from all children were simultaneously tested for endomysial antibodies, IgA tissue transglutaminase antibodies, and antibodies against tissue transglutaminase in complex with gliadin in the IgA and IgG class of immunoglobulins. The exact McNemar test, Wilcoxon test, and Spearman's correlation coefficient were used to analyze the data. Results We found a significant asymmetry of the tissue transglutaminase antibodies test compared with the antibodies against tissue transglutaminase neo-epitope test (P1). More patients (1.5%) had tissue transglutaminase an¬tibodies positive and antibodies against tissue transglutaminase neo-epitope negative results, whereas no patients had tissue transglutaminase antibodies negative and antibodies against tissue transglutaminase neo-epitope positive results. Of 59 children with tissue transglutaminase antibodies and/or endomysial antibodies positive results (P2), one (1.7%) did not have celiac disease. In agreement with the P1 study, four patients (6.8%) with confirmed celiac disease were tissue transglutaminase antibodies positive and antibodies against tissue transglutaminase neo-epitope negative. In this group, the sensitivity of the antibodies against tissue transglutaminase neo-epitope test for diagnosis of celiac disease was 91.4% (95% confidence interval, 81.0 - 97.1%). Among children diagnosed with functional gastrointestinal disorder (P3), all had negative serological test results, and none was diagnosed with celiac disease. Conclusions The results do not indicate that antibodies against tissue transglutaminase neo-epitope test would be an unambiguously better test than the currently used tissue transglutaminase antibodies.

The perception of celiac disease (CD) has changed dramatically during the last two decades, with the introduction of new serological tests with high specificity and sensitivity. CD prevalence in recent years is increasing as it is for other autoimmune diseases. Due to the high prevalence of CD, it is one of the most important diseases of the digestive tract. The clinical manifestations of CD have changed significantly in the last two decades, and these are much more diverse than reported previously. The new European Society for Pediatric Gastroenterology, Hepatology, and Nutrition diagnostic criteria for CD indicate that biopsy can be avoided in strictly selected groups of pediatric patients. The first experiences with these new criteria are positive. A meta-analysis evaluating the relationship between early nutrition and the risk of CD recommends that both early ( 7 months) introduction of gluten to the diet should be avoided, and that gluten should be introduced while the infant is still breastfeeding. The prevalence rates of CD between two cohorts with different infant feeding practice were significantly different, and the prevalence of CD in 12-year-old children were significantly reduced, suggesting that prevention of CD is possible.

This article was originally published with all author names incorrectly listed. All author names have now been transposed and appear correctly above. The original article was corrected.

Young age and thiopurine therapy are risk factors for lymphoproliferative disease among patients with inflammatory bowel disease (IBD). The aims of this study were to evaluate the prevalence of seropositivity for the Epstein–Barr virus (EBV) and human cytomegalovirus (CMV) among children and adolescents with IBD, to assess the viral load of EBV, CMV, and BK and JC polyomaviruses (BKV, JCV) in these patients, and to assess the influence of different therapeutic regimens on seroprevalence and viral load. Children who had been followed in our center were tested for EBV, CMV, BKV, and JCV in a cross-sectional study. One hundred and six children were included who had Crohn’s disease (68 %), ulcerative colitis (29 %), and unclassified IBD (3 %). We found that 64 % of patients were EBV seropositive. The proportion of EBV seropositive patients increased during childhood. Azathioprine therapy (p = 0.003) was associated with EBV seropositivity in a multiple logistic regression model, after adjusting for gender, age, and disease activity at determination. We found a significant association between the number of polymerase chain reaction copies and infliximab dose (p = 0.023). We did not find any significant association between CMV serology and CMV, BKV, or JCV viral load, or any other therapeutic regimen or clinical characteristics. Treatment with azathioprine appears to be a risk factor for early EBV seropositivity in children with IBD, and the infliximab dose was associated with a higher EBV viral load.

Prebiotics are used for stimulating the growth of beneficial microorganisms in the gut. However, it is very difficult to find a suitable prebiotic mixture that exclusively supports the growth of beneficial microbes such as bifidobacteria and lactobacilli. We tested the effects of a prebiotic mixture in vitro by incubating it with fecal samples and in vivo by administration of the prebiotic supplement to healthy adult volunteers, followed by analysis of their fecal microbiota. The effect of the oligosaccharides on bacterial metabolism was studied by analyzing short-chain fatty acid (SCFA) production in vitro and the SCFA pattern for the stool samples of volunteers. In the in vitro test, a higher proportion of bifidobacteria (25.77%) was seen in the total bacterial population after cultivation on a prebiotic mixture than on the control medium (7.94%). The gram-negative anaerobe count significantly decreased from 8.70 to 6.40 log CFU/g (from 35.21% to 0.60%) and the Escherichia coli count decreased from 7.41 to 6.27 log CFU/g (from 1.78% to 0.44%). Administration of a prebiotic mixture in vivo (9 g of galactooligosaccharides [GOS]+1 g of maltodextrins; daily for 5 days) significantly increased the fecal bifidobacterial count from 9.45 to 9.83 log CFU/g (from 40.80% to 53.85% of total bacteria) and reduced the E. coli count from 7.23 to 6.28 log CFU/g (from 55.35% to 45.06% of total bacteria). The mixture comprising GOS and maltodextrins thus exhibited bifidogenic properties, promoting the performance of bifidobacteria by boosting their growth and inhibiting the growth of undesirable bacteria.

Aim To develop and validate high-sensitive (hs) ELISA method for detection of adipophilin (adipose differentiation-related protein, ADRP) in human breast milk (BM) and to analyze adipophilin levels in BM during 12 months of lactation. Methods ADRP levels were determined using hsELISA method (Biovendor-Laboratory Medicine, Inc.) in colostrum (D0) and BM of 72 mothers was collected 1, 3, 6, and 12 months following delivery (M1, 3, 6, 12). Results ADRP was detectable in BM up to 12 months of lactation. Mean levels at D0 were 1.98 ± 0.12; M1, 2.83 ± 0.21; M3, 2.39 ± 0.17; M6, 2.57 ± 0.16; and at M12 3.25 ± 0.21 μg/ml. Significantly higher levels of ADRP were found in M1 and M12 when compared to D0 and in M12 when compared to M3 (overall P = 0.0001). No significant correlation was seen between ADRP levels in BM and adiponectin, body weight of infants, their birth length, body weight gain during the first year of life, or BMI of mothers before pregnancy. Conclusions We developed and validated hsELISA for detection of ADRP in human BM. ADRP was detectable in human BM during the whole 12 months of lactation period and its levels were intraindividually well-conserved.

The aim of this study was to assess the accuracy of serological tests in combination with clinical symptoms for diagnosing celiac disease (CD) according to the new proposed European Society for Pediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN) criteria. We retrospectively assessed children and adolescents aged 16 months –19 years who were examined for suspicion of CD (n = 345). Evaluation of clinical symptoms and the presence of tissue transglutaminase (anti-TG-IgA) and endomysial antibodies (EMA-IgA) as well as intestinal biopsies was performed in all patients. Human leukocyte antigens (HLAs) were not included. Among 345 biopsied children, 213 (62 %) children had anti-TG titers >10 times the upper limit of normal (ULN) and positive EMA antibodies. Ninety-nine (29 %) children also had symptoms suggestive of CD in addition to EMA positivity and elevated titers of anti-TG >10 times the ULN. In patients who were asymptomatic, but positive for EMA, and had anti-TG antibodies >10 times the ULN, the specificity of tests for Marsh 2–3 was only 85 %, while in symptomatic patients with the same antibodies levels, the specificity was 99 %. Conclusion: Our results reveal that intestinal biopsies could be omitted in 28 % of patients when the new ESPGHAN guidelines are applied. Due to high accuracy of serological tests in combination with clinical symptoms for diagnosis of CD, the new guideline seems to be applicable even without the use of HLA testing.

Five strains of bifidobacteria of human origin and 2 strains of bifidobacteria of animal origin were tested for growth in 10 samples of human milk. Growth of Bifidobacterium bifidum in human milk was accompanied by a decrease in pH and production of acids. The number of viable cells of Bifidobacterium animalis was decreased from 6 log cfu mL −1 to 3 log cfu mL −1 after incubation in human milk. There were significant differences ( P B. bifidum and B. animalis in milk samples tested. Resistance to lysozyme and the ability to utilise human milk oligosaccharides (HMOs) were identified as the most important factors affecting the growth of bifidobacteria in human milk. Four out of 5 strains of human origin were resistant to lysozyme and utilised HMOs. In contrast, B. animalis was susceptible to lysozyme and did not utilise HMOs.

Background: The role of the IBD5 locus in development of Crohn's disease (CD) has not been clarified. In the Czech population we examined its genetic association using variants of the SLC22A4 (rs1050152), SLC22A5 (rs2631367), two single nucleotide polymorphisms (SNPs) shown to be associated with CD in genome-wide studies (rs6596075 and rs2188962), and four SNPs previously shown to tag the haplotype blocks 4, 7, 9, 10 of the IBD5 locus (IGR2063b_1, IGR2230a_1, IGR100Xa_1, IGR3236a_1). Methods: The genotype, phenotype, and allelic frequencies were compared between 469 unrelated patients with CD (177 pediatric-onset, 292 adult-onset) and 470 unrelated healthy controls, all Caucasians of Czech ancestry. Results: The most significant difference between patients and controls was found for the SNP rs6596075 (odds ratio [OR] = 0.70 for the G allele; 95% CI 0.52–0.94) in the dominant model and SNP IGR2063b_1 (OR = 1.38 for the G allele; 95% CI 1.14–1.67) in the log-additive model. We found a strong linkage disequilibrium across the IBD5 locus except rs6596075. The haplotype consisting of minor alleles of all tested SNPs except rs6596075 was carried by 31% patients and 23% control subjects (OR = 1.35, 95% CI 1.06–1.72). The association of variants in SLC22A4 and SLC22A5 was dependent on this risk haplotype, while the strong association of the rs6596075 was seemingly independent. In the analysis of subphenotypes we found only an association of the penetrating disease with rs6596075 (OR = 2.13; 95% CI 1.31–3.47). Conclusions: Our study confirms the importance of IBD5 in determining CD susceptibility, and demonstrates that two independent genetic factors may be responsible for the association observed within this locus. (Inflamm Bowel Dis 2010)

Oats in a gluten-free diet increase the diet's nutritional value, but their use remains controversial. Contamination with prolamins of other cereals is frequent, and some clinical and experimental studies support the view that a subgroup of celiac patients may be intolerant to pure oats. Thus, this issue is more complex than previously suggested. In order to produce oats that are safe for all celiac patients, the following topics should be addressed: selection of oat cultivars with low avenin content, research on such recombinant varieties of oats, development of assay methods to detect avenins in oat products, guidelines for the agricultural processing of oats and the manufacture of oat products, as well as guidelines for following up with celiac patients who consume oats.

Background and aims: Wilson disease (WD) is an inherited disorder of copper metabolism. When treated, the outcome can be excellent, although the long-term survival has yet to be well documented. The aim of this study was to describe the long-term outcome of a cohort of patients with WD and to assess those factors affecting the phenotypic manifestation of WD. Methods: The presence of mutations to the ATP7B gene, the clinical manifestations, treatments and the long-term outcomes were analysed retrospectively in 117 patients with WD (59 men and 58 women, aged at evaluation 38.5 ± 11, range 16–63 years). Results: Fifty-five patients with a neurological presentation, 51 patients with a hepatic presentation and 11 asymptomatic patients were followed up for an average of 15.1 ± 10 years (median 12 years, range 1–41 years). The H1069Q ATP7B gene mutation was the most frequent genetic variant (54.3%); the frequency of this mutation did not differ between patients with either the hepatic or the neurological presentation (P=0.099). d-penicillamine or zinc salts (81 and 17% respectively) were used for treatment, and three patients underwent liver transplantation. The majority of symptomatic patients became asymptomatic, or improved, during the follow-up (82% patients with hepatic presentation, 69% with neurological presentation). The long-term survival of patients with WD did not differ from that of the general Czech population (P=0.95). Conclusions: Long-term follow-up shows a satisfactory response in the great majority of adequately treated patients with WD and survival coincides with that of the general population.

Aim: The aim of our study was to analyse cytokine composition of human milk and its relationship to the development of eosinophilic colitis (EC). Methods: Cytokines were measured by ELISA method in breast milk of 20 mothers of infants who developed EC and 20 controls. Results: We found significantly higher concentrations of interferon-gamma (IFN-gamma) (Th1 cytokine) in breast milk received by EC infants compared to controls (p = 0.0004). In contrary, IL-18 (Th1-inducing cytokine) was significantly higher in breast milk received by healthy infants comparing to EC infants (p = 0.001). Regulatory cytokine transforming growth factor beta1 (TGF-beta1) showed higher concentrations in breast milk received by healthy infants, although the difference from EC group was not significant (p = 0.072). Conclusion: The results of our study showed that infants with EC were receiving breast milk with a possibly risky cytokine pattern indicating cytokine imbalance, impaired immunoregulation and the early Th1 shift.

Objectives An association of variants in the genes encoding the interleukin 23 receptor (IL23R, p.Arg381Gln, rs11209026), and the autophagy-related gene 16-like 1 (ATG16L1, p.Ala197Thr, rs2241880) with Crohn disease (CD) was identified by whole genome association studies, and subsequently confirmed by other works. The aim of this study was to assess this association in the Czech population. Subjects and methods In a case-control study 333 patients with CD (137 paediatric and 196 adult-onset) and 499 unrelated healthy controls were genotyped using TaqMan SNP assays. Results The IL23R p.381Gln allele was protective against CD in the Czech population (allelic frequency 3.2% in patients vs 5.5% in control subjects; OR 0.56, 95% CI 0.33-0.93, P=0.02). ATG16L1 p.197Ala allele conferred increased risk of CD (allelic frequency 60% in patients vs 51% in controls; OR 1.25, 95% CI 1.02-1.52, P=0.03). There was no appreciable difference in the effect of the associated alleles across the strata of CARD15-conferred risk. The IL23R and ATG16L1 variants did not influence the age at diagnosis, and in the genotype-phenotype analysis, the only detected association was a weak one between IL23R p.381Gln and involvement of the upper gastrointestinal tract (uncorrected P=0.031). Conclusions We confirmed the role of IL23R and ATG16L1 in the CD susceptibility in the Czech population, and found a weak protective effect of IL23R p.381Gln against upper gastrointestinal tract involvement.

Intestinal microbiota in exclusively breast-fed infants with blood-streaked stools and in healthy exclusively breast-fed babies was compared. Total anaerobes, bifidobacteria, lactobacilli, coliform bacteria, enterococci and clostridia were quantified by cultivation methods in feces of 17 full-term exclusively breastfed patients (aged 16.3 +/- 7.4 weeks) with blood-streaked stools and in the control group of 22 healthy fullterm exclusively breast-fed infants (13.7 +/- 6.4 weeks). Specific fluorescence in situ hybridization kits for Bifidobacterium spp. were used for the quantitative detection of bifidobacteria in samples. Control samples had significantly (p0.05) higher counts of total anaerobes. Bifidobacteria were not detected in patients' samples in 65 % and in controls in 36 % (p0.01). Bifidobacteria counts were also significantly higher in the control group (p0.01). Furthermore, clostridia strains were detected only in feces from bifidobacteria-negative infants reaching counts8 log CFU/g. Lactobacilli were not detected in 65 % patients and in 45 % control samples. However, this difference was not significant as well as the difference in lactobacilli counts. Eosinophilia was observed in 35 % of patients, low IgA concentration in 71 % and also low IgG concentration in 71 %. pANCA positivity was found in 53 % of patients. In conclusion a significant low proportion of bifidobacterial microbiota in patients with blood-streaked stools was shown in comparison with controls.

Background: Breastfeeding may protect children from developing metabolic syndrome and other diseases later in life. We investigated novel proteins in human breast milk that might play a role in this process.Methods: We used ELISA to measure adiponectin, adipocyte and epidermal fatty acid binding proteins (AFABP, EFABP), and leptin concentrations in human breast milk obtained from 59 mothers 48 h after initiation of lactation. Using a questionnaire and medical records, we collected information about the mothers and newborns.Results: Mean (SE) adiponectin concentrations in breast milk were 13.7 (0.8), range 3.9–30.4 μg/L; AFABP concentrations 26.7 (4.4), range 1.2–137.0 μg/L; EFABP concentrations 18.1 (1.4), range 0.8–47.0 μg/L; and leptin concentrations 0.50 (0.05), range 0–1.37 μg/L. We found a significant correlation between AFABP and EFABP concentrations (r = 0.593, P Conclusions: Concentrations of adiponectin, AFABP, and EFABP in human breast milk are related to nutritional variables of mothers and newborns and thus may play a role in the protective effects of breastfeeding.

Sixty-four breast-fed infants and 23 calves were investigated for bacteria and enzymatic activity in their faecal samples. The bacteria were measured using cultivation and fluorescence in situ hybridization. Enzymatic activity was also examined. Forty-seven (64%) infants and all the calves had high numbers of bifidobacteria (usually >9 log CFU g−1) in their faeces, but 17 infants (36%) did not have a detectable amount of the bacteria. Most of the bifidobacteria-negative infants had significant quantities of clostridia in their faecal flora. While the infants did not have significantly higher counts of bifidobacteria, the samples from calves contained significantly (P

Background The aim of this study was to assess the pediatric population that suffered from inflammatory bowel disease (IBD) in the Czech Republic and to determine the incidence of Crohn disease (CD) in children up to 15 years age between 1990 and 2001. Methods Diagnostic criteria for CD, ulcerative colitis (UC), and indeterminate colitis (IC) were defined. Medical records provided a source of basic information about the children. A standardized protocol was filled out and sent to the coordinator of the study. All protocols were checked to see whether the data corresponded to the defined criteria and then were processed further. The study was retrospective in character for the years 1990 to 1999 and prospective for the years 2000 and 2001. Results Diagnostic criteria were met in 470 patients with IBD; 201 of them turned 18 years old during the study period. CD was diagnosed in 223 patients. The incidence of CD in children up to 15 years of age increased from 0.25/100,000 in 1990 to 1.25/100,000 in 2001. Eighty-two percent of children with CD were treated with aminosalicylates in combination with corticosteroids; 29% of patients received azathioprine. Severe growth retardation was recorded in 6.4% of adolescents with CD at the age of 18. UC was diagnosed in 202 patients. Therapy with aminosalicylates only was sufficient for control of the disease in 23% patients; 68% children were treated with corticosteroids, 15 of them (23% of the whole group) received additional azathioprine. Criteria for IC were met in 9.8% of all patients with IBD. Conclusion This study confirmed an increase in incidence of CD in children younger than 15 years in the Czech Republic.

Interferon-γ release assay (IGRA) is widely used for screening of latent tuberculosis (TB) before and during biological therapy (BT). An indeterminate result of IGRA represents a limitation in the management of inflammatory bowel disease (IBD). Data on factors influencing IGRA results are scarce in children. The aim of the study was to identify factors influencing IGRA results in children with IBD.Seventy-two children with IBD (59 Crohn disease, 11 ulcerative colitis, 2 IBD-unclassified) indicated for BT were tested for TB infection (history, TB skin test, chest radiograph, IGRA; QuantiFERON-TB Gold in tube [QFT]) and consecutively retested using QFT in 1-year intervals.We recorded 165 results of QFT (3% positive, 87% negative, and 10% indeterminate results). During follow-up we identified 4 conversions of negative QFT to positivity (3%) and 4 reversions (4%). Patients with indeterminate results of QFT had significantly lower actual weight-for-height z score (P = 0.022), higher platelet count (P = 0.00017), and lower levels of serum albumin (P = 0.015) compared with patients with positive or negative QFT. Indeterminate QFT was associated with corticosteroid treatment, BT, and disease activity, but not with treatment by immunomodulators. In a subanalysis of patients with Crohn disease alone, Pediatric Crohn's Disease Activity Index was identified as single independent risk factor for indeterminate results (P = 0.00037).Although corticosteroid treatment is traditionally considered to be the main risk factor for indeterminate results of IGRA, the disease activity of IBD has even more profound effects on the results.

AIM: To develop and validate high‐sensitive (hs) ELISA method for detection of adipophilin (adipose differentiation‐related protein, ADRP) in human breast milk (BM) and to analyze adipophilin levels in BM during 12 months of lactation. METHODS: ADRP levels were determined using hsELISA method (Biovendor‐Laboratory Medicine, Inc.) in colostrum (D0) and BM of 72 mothers was collected 1, 3, 6, and 12 months following delivery (M1, 3, 6, 12). RESULTS: ADRP was detectable in BM up to 12 months of lactation. Mean levels at D0 were 1.98 ± 0.12; M1, 2.83 ± 0.21; M3, 2.39 ± 0.17; M6, 2.57 ± 0.16; and at M12 3.25 ± 0.21 μg/ml. Significantly higher levels of ADRP were found in M1 and M12 when compared to D0 and in M12 when compared to M3 (overall P = 0.0001). No significant correlation was seen between ADRP levels in BM and adiponectin, body weight of infants, their birth length, body weight gain during the first year of life, or BMI of mothers before pregnancy. CONCLUSIONS: We developed and validated hsELISA for detection of ADRP in human BM. ADRP was detectable in human BM during the whole 12 months of lactation period and its levels were intraindividually well‐conserved.

Human milk (HM) contains as the third most abundant component around 200 different structures of human milk oligosaccharides (HMOs). HMOs are the first and irreplaceable prebiotics for infants, supporting bifidobacteria as the most important bacterial group in an infant intestine. The aim of our study was to test the growth of bifidobacteria in HM and on HMOs. Bifidobacteria were isolated from two groups of infants. The first one (eight strains) were isolated from infants who had bifidobacteria in their feces but, after a short period of time (4 to 24 days), bifidobacteria were no longer detected in their feces (disappeared bifidobacteria [DB]). The second group of bifidobacteria (eight strains) originated from infants with continual presence of bifidobacteria in their feces (persistent bifidobacteria [PB]). There were significant differences (p

Adiponectin, adipocyte fatty acid-binding protein (AFABP), and leptin have been shown to be present in human breast milk (BM). We determined intraindividual changes of BM levels of these proteins during 12 months of lactation.Proteins were measured using a high-sensitivity enzyme-linked immunosorbent assay method in 72 healthy mothers after delivery (day 0, D0) and after 1, 3, 6, and 12 months of lactation.Adiponectin levels in BM on D0 were 22.8 ± 0.8 (mean ± standard error of the mean), in 1 month (M1) 22.0 ± 0.6, in 3 months (M3) 20.5 ± 0.6, in 6 months (M6) 21.4 ± 0.8, and in 12 months (M12) 25.7 ± 1.4 ng/mL. AFABP levels were 12.3 ± 2.0, 6.2 ± 1.3, 1.3 ± 0.2, 2.5 ± 1.0, and 4.6 ± 1.9 ng/mL, respectively. Leptin levels were 0.3 ± 0.04, 0.2 ± 0.03, 0.1 ± 0.01, 0.1 ± 0.02, and 0.2 ± 0.04 ng/mL, respectively. We found significantly higher levels of adiponectin in M12 in comparison to M3 and M6 (P = 0.0026), higher levels of AFABP in D0 and M1 when compared with M3, M6, and M12 (P0.0001), and higher levels of leptin on D0 than in M1, M3, M6, and M12 (P0.0001). AFABP levels correlated negatively with infants' body weight in M1, but there was no correlation throughout the lactation period between body weight and other proteins. We found positive correlation between adiponectin, AFABP, and leptin throughout the lactation.All of the hormones were detectable in BM up to 12 months of lactation, with decreasing trend until M3 and subsequent increase till M12. We speculate that higher levels in M6 and M12 may be caused by longer intervals between breast-feeding due to the introduction of complementary food.

Wilson disease (WD) is an inherited disorder of copper metabolism. When treated, the outcome can be excellent, although the long-term survival has yet to be well documented. The aim of this study was to describe the long-term outcome of a cohort of patients with WD and to assess those factors affecting the phenotypic manifestation of WD.The presence of mutations to the ATP7B gene, the clinical manifestations, treatments and the long-term outcomes were analysed retrospectively in 117 patients with WD (59 men and 58 women, aged at evaluation 38.5 ± 11, range 16-63 years).Fifty-five patients with a neurological presentation, 51 patients with a hepatic presentation and 11 asymptomatic patients were followed up for an average of 15.1 ± 10 years (median 12 years, range 1-41 years). The H1069Q ATP7B gene mutation was the most frequent genetic variant (54.3%); the frequency of this mutation did not differ between patients with either the hepatic or the neurological presentation (P = 0.099). d-penicillamine or zinc salts (81 and 17% respectively) were used for treatment, and three patients underwent liver transplantation. The majority of symptomatic patients became asymptomatic, or improved, during the follow-up (82% patients with hepatic presentation, 69% with neurological presentation). The long-term survival of patients with WD did not differ from that of the general Czech population (P = 0.95).Long-term follow-up shows a satisfactory response in the great majority of adequately treated patients with WD and survival coincides with that of the general population.

Objective: Orexin A (OXA) is a hypothalamic neuropeptide involved in regulation of food intake and nutritional status. There are multiple disturbances of neuropeptide signaling described in girls with anorexia nervosa (AN), but OXA levels have not been addressed in this population to date. Therefore, we analyzed OXA levels of AN girls in this study. Method: OXA (radioimmunoassay/RIA/method), leptin, insulinlike growth factor-1 (IGF-1), and insulinlike growth factor-1 binding protein-3 (IGFBP-3) levels were measured before and after 8 weeks of realimentation in 36 girls with AN and in 14 healthy controls (control group: CG). Results: Average weight increased significantly in AN during the study (p < .0001), while plasma levels of OXA decreased (before realimentation: 56.2 ± 2.4 pg/ml; after realimentation: 47.5 ± 1.4 pg/ml; p = .0025). OXA levels before realimentation differed from levels in the CG (47.15 ± 2.6 pg/ml, p = .034), but not afterward. We did not find any correlation between OXA and age, height, weight, BMI; or IGF-1, IGFBP-3, and leptin levels. Discussion: OXA levels in untreated AN patients differ significantly from healthy subjects and decrease during realimentation. These findings indicate that OXA may be involved in the nutritional regulation of malnourished children and adolescents. © 2010 by Wiley Periodicals, Inc. Int J Eat Disord 2011

The aim of our study was to analyse cytokine composition of human milk and its relationship to the development of eosinophilic colitis (EC).Cytokines were measured by ELISA method in breast milk of 20 mothers of infants who developed EC and 20 controls.We found significantly higher concentrations of interferon-gamma (IFN-gamma) (Th1 cytokine) in breast milk received by EC infants compared to controls (p = 0.0004). In contrary, IL-18 (Th1-inducing cytokine) was significantly higher in breast milk received by healthy infants comparing to EC infants (p = 0.001). Regulatory cytokine transforming growth factor beta1 (TGF-beta1) showed higher concentrations in breast milk received by healthy infants, although the difference from EC group was not significant (p = 0.072).The results of our study showed that infants with EC were receiving breast milk with a possibly risky cytokine pattern indicating cytokine imbalance, impaired immunoregulation and the early Th1 shift.

Twenty-eight exclusively breast-fed healthy infants and 16 infants also exclusively breast-fed with allergic colitis (aged 85 +/- 60 and 98 +/- 58 d, respectively) were screened for differences in fecal flora. Bifidobacteria were detected in 23 healthy infants and only in 4 fecal samples of infants with allergic colitis. All bifidobacteria-free infants possessed Gram-positive regular rods as a major group of their fecal flora. These bacteria were identified as clostridia using genus-specific FISH probe. Infants with allergy colitis possessed significantly lower counts of bifidobacteria and total anaerobes and significantly higher counts of clostridia in their feces. In healthy infants, Bifidobacterium longum was the most frequently found species (54.5% of the samples), followed by B. adolescentis (20.0), B. breve (18.2), B. bifidum (16.4), B. dentium (10.9) and B. pseudocatenulatum (1.80). Bifidobacterial isolates from two babies with allergic colitis were identified as B. longum, one child from patients group contained species B. dentium and one baby B. adolescentis. Our results suggest that there are significantly lower counts of bifidobacteria in infants with allergic colitis than in healthy infants.

Background The CTLA4 (cytotoxic T-lymphocyte antigen 4) gene is associated with several immunopathologic diseases and because of its important immuno-regulatory role it may be considered also a plausible candidate for a genetic association with inflammatory bowel diseases. Previously published studies found no association of CTLA4 with Crohn's disease itself, but some indicated an association with its subphenotypes. The aim of this study was to assess the association in the Czech population, using a set of markers shown to associate with other diseases. Methods Six polymorphisms within the CTLA4 region were investigated in 333 patients with Crohn's disease and 482 unrelated healthy controls, all Caucasians of Czech origin. The genotypes of the SNPs were determined using the TaqMan SNP genotyping assays. Haplotypes were reconstructed using an expectation-maximization algorithm, and their association with the condition was assessed using log-linear modeling. Then, potential interactions were tested between the CTLA4 variants and other genetic factors known to confer the disease susceptibility. Results No crude associations with Crohn's disease were found for the tested CTLA4 variants under the log-additive or dominant models. However, when stratified for the genetic risk conferred by the variants in the NOD2 (the p.Leu1007fsX1008, rs5743293) or the IL23R (p.R381Q, rs11209026), a significant negative association emerged for the minor alleles of CTLA4 CT60 (rs3087243), JO31 (rs11571302), JO27-1 (rs11571297) polymorphisms. This negative association with CTLA4 was apparent only in the strata defined by presence minor alleles at the NOD2 rs5743293 (here the CTLA4 CT60 A coffered an OR = 0.43, 95%CI 0.19 - 0.95 for the presence of CT60 A), or IL23R rs11209026 (here the OR for presence of CT60 A was 0.23, 95%CI 0.07 - 0.71). We observed this effect also for the haplotype consisting of minor alleles of the three tightly linked CTLA4 markers. Furthermore, this haplotype was associated with the younger age at diagnosis (OR 1.52, 95%CI 1.09 - 2.11, p = 0.014). Conclusions A protective effect of a CTLA4 haplotype was unmasked after stratification for the risk variants in the NOD2 and IL23R genes, and may point towards the biological relevance of the molecule in the pathogenesis of the disease.

The existence of a significant linear relationship between the concentration of chlorides and the activity of pepsinogen (PG) in the urine was ascertained in the case of full-term infants 3 days to 6 weeks of age. At the age of 4-6 weeks, a significant relationship was found between the urinary pepsinogen activity and the urinary creatinine concentration, and between the urinary pepsinogen activity in the urine and the urine osmolality. Immediately after birth, the Pg7 fraction of PG II in the urine was found in all cases and, at the age of 4-6 weeks, in 11% of cases. In regard to the time factor, the conspicuous drop in the occurrence of the Pg7 fraction corresponds to the new qualitative relationship between the pepsinogen activity in the urine and the creatinine concentration in the urine and between the former and the urine osmolality. In premature infants, the Pg7 fraction disappears more slowly. The spectrum of pepsinogens in the urine was examined in children suffering from various diseases. In a girl with lymphoma, we found the Pg7 fraction, but this finding was temporary only.