First case report of rare congenital adrenal insufficiency caused by mutations in the CYP11A1 gene in the Czech Republic.

We characterized a case of congenital adrenal insufficiency caused by cholesterol side-chain cleavage enzyme (P450scc) deficiency. The patient presented after birth with cardiopulmonary instability, hyponatremia, hyperkalemia, hypoglycemia and metabolic acidosis. We confirmed primary adrenal insufficiency. There were no signs of the external genitalia virilism. The replacement therapy with glucocorticoids and mineralocorticoids led to normal laboratory results. At the age of 12 years, we confirmed hypergonadotropic hypogonadism, which revealed disorder of steroidogenesis in the adrenal glands and in the gonads. The enzymatic block was found at the beginning of steroidogenesis. The mutation was confirmed in the CYP11A1 gene. The patient is compound heterozygote for the novel CYP11A1 missense mutation c.412G>A (p.Gly138Arg) in exon 2 and frameshift mutation c.508_509delCT (p.Leu170Valfs*30) in exon 3. The CYP11A1: c.412G>A (p.Gly138Arg) was predicted as pathogenic by in silico analysis. So far, only 19 patients with CYP11A1 mutations causing P450scc deficiency have been reported worldwide. There are no related reports in the Czech Republic.

Parameters of oxidative stress, DNA damage and DNA repair in type 1 and type 2 diabetes mellitus.

OBJECTIVES: (i) to determine the extent of oxidative stress and DNA damage and repair using a panel of selected markers in patients with type 1 and type 2 diabetes mellitus (T1DM, T2DM), (ii) to find their possible relationships with diabetes compensation and duration, and finally (iii) to test for the effect of functional polymorphisms in the 8-oxoguanin DNA glycosylase (rs1052133), catalase (rs1001179) and superoxide dismutase (rs4880) genes on respective intermediate phenotypes. METHODS: A total of 207 subjects (23 children and 44 adults with T1DM, 52 adult patients with T2DM and 88 healthy adult control subjects) were enrolled in the study. The following markers of redox state were determined in participants: erythrocyte superoxide dismutase (Ery-SOD), whole blood glutathione peroxidase (WB-GPx), erythrocyte glutathione (Ery-GSH), plasma total antioxidant capacity (P-tAOC) and plasma malondialdehyde (P-MDA). Furthermore, the extent of DNA damage and repair was ascertained using the following parameters: DNA single strand breaks (DNAssb), DNA repair capacity (DNArc) and DNA repair index (DNRI). RESULTS: Comparison of T1DM vs. T2DM patients revealed significantly higher Ery-GSH content (P < 0.0001) and significantly lower Ery-SOD activity (P = 0.0006) and P-tAOC level (P < 0.0001) in T1DM subjects. T2DM diabetics exhibited a significant increase in DNAssb (P < 0.0001) and significant decrease in both DNArc (P < 0.0001) and DNRI (P < .0001) compared with T1DM patients. Patient's age (irrespective of DM type) significantly correlated with DNAssb (r = 0.48, P < 0.0001), DNArc (r = -0.67, P < 0.0001) and DNRI (r = -0.7, P < 0.0001). Allele frequencies of all studied polymorphisms did not exhibit any significant association with the investigated parameters. CONCLUSION: We demonstrated significant age- and DM type-related changes of oxidative DNA modification and capacity for its repair in subjects with T1DM and T2DM.

Evaluation of faecal calprotectin as a valuable non-invasive marker in distinguishing gut pathogens in young children with acute gastroenteritis.

AIM: The aim of the study is to evaluate faecal calprotectin (f-CP) in children ≤3 years of age with acute gastroenteritis (AG) as an early predictor of bacterial inflammation. METHODS: We prospectively analysed f-CP levels and diagnostic workup in 107 consecutive children (66 AG, 41 controls). RESULTS: Children with bacterial AG (BAG) was found to have higher diarrheal frequency (p < 0.01), fever (p < 0.01), erythrocyte sedimentation rate (p < 0.001), white blood count (p < 0.01) and C-reactive protein (CRP) (p < 0.001) compared with viral AG (VAG). Vomiting was frequent in VAG (p < 0.001). f-CP negatively correlated with age in controls (r = -0.5998). BAG demonstrated significantly higher f-CP levels [median, 219 µg/g, interquartile range (IQR): 119-350.2] compared with VAG (49.3 µg/g, IQR: 8.8-131.1) as well as controls (26.5 µg/g, IQR: 14.9-55.1) (p < 0.001). VAG and control f-CP levels were similar. f-CP was the best-rated marker of BAG with a diagnostic accuracy of 92%. Receiver-operator characteristic analysis revealed an area under curve of 0.95 for identifying BAG; sensitivity and specificity of f-CP were 93% and 88%, respectively, at an adjusted cut-off point of 103.9 µg/g faeces. Combined f-CP and CRP yield improved diagnostic accuracy of 94% for BAG. CONCLUSION: f-CP facilitates early discrimination between bacterial and viral causes of AG in young children. Combining f-CP with CRP increases the diagnostic power of diagnosing BAG.

Epidemiology of Helicobacter pylori infection in asymptomatic children: a prospective population-based study from the Czech Republic. Application of a monoclonal-based antigen-in-stool enzyme immunoassay.

BACKGROUND: Acquisition of Helicobacter pylori occurs mainly in childhood and is significantly influenced by geographical variations. The aim of this study is to evaluate the prevalence of H. pylori infection in a population-based sample of asymptomatic children in the Czech Republic. Furthermore, this study aims to identify potential risk factors associated with this infection. MATERIALS AND METHODS: A prospective, cross-sectional, population-based study was undertaken in 1545 asymptomatic Czech children (aged 0-15 years; male 49.3%). Active H. pylori infection was diagnosed by monoclonal antibody-based antigen-in-stool enzyme immunoassay. Socio-demographic details of each subject were analyzed using a self-administered standardized questionnaire. Multiple regression analysis was performed. RESULTS: Overall, 7.1% of asymptomatic children were diagnosed with H. pylori infection. Of the infected children, 5.8% lived in the general population. A positive association was found with increasing age, although not with gender. Independent risk factors associated with H. pylori infection in our pediatric population were: the number of children in a household (odds ratio [OR] 4.26; confidence interval [CI] 1.91-9.80); lack of formal education of fathers (OR 0.23; CI 0.18-0.64) and institutionalized children (OR 6.33; CI 2.25-26.50). CONCLUSIONS: This study of a large cohort of children demonstrated that, independent of gender, H. pylori infection in the Czech Republic is among the lowest reported in Europe. Socioeconomically disadvantaged children, unfortunately, are still at risk of harboring this potentially preventable infection in this low-prevalence region.

Cytokine tumor necrosis factor-alpha A promoter gene polymorphism at position -308 G-->A and pediatric inflammatory bowel disease: implications in ulcerative colitis and Crohn's disease.

OBJECTIVES: Our pilot study aimed to determine the effect of tumor necrosis factor-alpha (TNF-alpha) 308 G-->A promoter single-nucleotide polymorphism in pediatric inflammatory bowel disease (IBD), its influence on inflammatory activity and the clinical manifestations. METHODS: We obtained genomic DNA from 164 subjects, 82 with long-standing IBD aged 8 to 18 years: 46 with Crohn disease (CD) and 36 with ulcerative colitis (UC). Eighty-two healthy children served as the control population. Genotyping was determined by using a restriction enzyme-based assay. TNF-alpha 308 G-->A polymorphism was assessed in terms of inflammatory (C-reactive protein [CRP]) and disease activity. The latter was assessed by the Pediatric Crohn's Disease Activity Index (PCDAI) and the Truelove index for CD and UC, respectively. RESULTS: Significant differences in TNF-alpha 308 A polymorphism were found between the IBD group and controls (P < 0.05) and the UC group and controls (P < 0.001). No differences were noted between TNF-alpha 308 A polymorphism and clinical characteristics in UC. The frequency of the -308 A allele of TNF was not different in CD compared with that in the control group. The frequency of TNF-alpha 308 A genotype was significantly higher in CD patients with predominantly stenosing/penetrating disease compared with patients without complications (P < 0.001) and healthy controls (P < 0.01). In CD patients, those carrying TNF -308 A had a significant increase in CRP (P < 0.05) and the PCDAI (P < 0.05). In CD, CRP levels strongly correlated with the PCDAI (r = 0.6150, P < 0.001). In UC, significant differences among the mean levels of CRP (P < 0.05) and disease activity (P < 0.001) related to TNF-alpha 308 A polymorphism were found. Allele distribution (odds ratio, 12.9; CI, 1.18-140.81, P < 0.001) and CRP serum levels (odds ratio, 1.020; CI, 1.00-1.04, P < 0.001) were independently associated with CD complications. CONCLUSIONS: Although not necessarily dictating IBD initiation, the TNF-alpha 308 A polymorphism may play a role in modifying the CD phenotype. The polymorphism may influence disease activity as well as more intense inflammatory activity in both forms of IBD and may modify the progression of chronic digestive tract inflammation.

Effects of a specially designed fermented milk product containing probiotic Lactobacillus casei DN-114 001 and the eradication of H. pylori in children: a prospective randomized double-blind study.

GOALS: To determine the efficacy of triple therapy supplemented with a specially designed fermented milk product containing specific probiotic Lactobacillus casei (L. casei) DN-114 001 strain on Helicobacter pylori eradication in children. BACKGROUND: Lactobacillus species possess in vitro activity against H. pylori. There are no consistent data on the impact of eradication therapy supplemented with probiotics on H. pylori cure rates in childhood in vivo. STUDY: Multicenter, prospective, randomized, double-blind controlled study. Eighty-six symptomatic H. pylori-positive children were randomized either to receive the control treatment of omeprazole, amoxicillin, and clarithromycin (OAC) for 7 days or the test treatment of omeprazole, amoxicillin, and clarithromycin for 7 days supplemented with fermented milk (Actimel) containing L. casei DN-114 001 (OAC-LC), for 14 days. H. pylori status was assessed at 4 weeks following therapy using two noninvasive tests. RESULTS: Intention-to-treat (ITT) based eradication rates for the OAC-LC group were 84.6% (95% CI, 71.2%-95.5%), and 91.6% (95% CI, 76.9%-98.2%) by per-protocol (PP) analysis. Eradication in the OAC group was 57.5% (95% CI, 42.2%-72.3%) in the ITT set and 61.3% (95% CI, 44.4%-75.0%) in the PP group. Eradication success was higher in the OAC-LC group compared with the OAC group in both ITT (P=0.0045) and PP analysis (P=0.0019). Primary resistance for clarithromycin could be determined in 21.2%. Side effects were infrequent. Drug compliance was good throughout the study. CONCLUSION: Supplementation with fermented milk, containing live special probiotic L. casei DN-114 001, confers an enhanced therapeutic benefit on H. pylori eradication in children with gastritis on triple therapy.

Aspects of oxidative stress in children with type 1 diabetes mellitus.

Diabetes mellitus is considered to be one of a rank of free radical diseases. The existence of hyperglycemia produces increased oxidative stress (OS) via non-enzymatic glycation, glucose autoxidation, and alterations in polyol pathway activity with subsequent influences on the whole organism. In childhood, Type 1 diabetes prevails and is characterized by its autoimmune character with progressive destruction of beta cells and lack of insulin in genetically predisposed patients. Our study focused on diabetic children and their 1st degree relatives and confirmed increased oxidative stress in diabetic children as well as a similar tendency in their siblings. Following this, we carried out a one-year study comprising diabetic children supplemented with vitamins E and C. The vitamin treatment improved diabetes control and reduced markers of oxidative stress substantially when compared with non-supplemented diabetic children. As oxidative stress impairs not only lipids and proteins, but also DNA, we attempted to examine the level of DNA strand breaks as well as DNA repair processes using comet assay modifications. Though children with Type 1 diabetes demonstrated increased oxidative stress (lower SOD and GSH when compared with healthy children), their oxidative DNA damage (measured as DNA strand breaks) were not substantially altered compared with normals. On the other hand, their DNA repair capacity was significantly increased. This demonstrates a stimulated DNA repair process that is most certainly a response to the permanently elevated state of oxidative stress. Owing to the presented results, it is appropriate to ponder the increased influence of oxidative stress on children with Type 1 diabetes and to take into account this fact when considering their treatment.

Acquired angioedema and Helicobacter pylori infection in a child.

UNLABELLED: We describe a 10-year-old boy with acquired Helicobacter pylori infection and simultaneous angioedema which is a rare but life-threatening condition. Our patient was hospitalised with generalised angioedema and severe circulatory shock due to extreme loss of fluids and proteins into interstitial tissues (weight gain 10 kg within 2 days, extreme haemoconcentration--haemoglobin 206 g/l, haematocrit 0.570, leucocytosis 18,300 /microl, high lactate 13.8 mmol/l) and simultaneous failure of the complement system (C3 <0.16 g/l, C4 <0.13 g/l, CH50 45 U/ml, i.e. 50% of normal value, C1 inhibitor 0.21 g/l at the lower limit). All possible known causes of angioedema were excluded (infection, allergy, auto-immune disease, NSAIDs, lymphoproliferative disease) except for the simultaneous H. pylori infection which was proven serologically and histologically. Eradication therapy led to a complete remission of the H. pylori infection. An absence of angioedema and the restoration of the complement system was later observed. To the best of our knowledge, no similar case report of a child has yet been published. CONCLUSION: Helicobacter pylori infection should be considered in the development of angioedema in childhood.

Parameters of oxidative stress in children with Type 1 diabetes mellitus and their relatives.

Oxidative stress (OS) plays an important role in the pathogenesis of Type 1 diabetes mellitus (DM). The aim of the study was to compare OS parameters in diabetic children and their first-degree relatives. Fifty diabetic children from the West Bohemian Region were examined as well as their 32 siblings (12 Boys and 20 girls) and 65 of their parents during a period of 6 months. Thirty healthy sex- and age-matched children studied before planned surgeries were normal controls for children, 40 healthy adult volunteers were controls for parents. OS parameters were evaluated in all participants of the study (superoxide dismutase, SOD; glutathione peroxidase, GSHPx; plasma antioxidant capacity, AOC; reduced glutathione, GSH; and malondialdehyde, MDA) and also Type 1 DM-associated antibodies (ICA and GADA). The results in diabetic children showed significantly lower GSHPx and AOC and increased MDA when compared with healthy children. Similar findings were found in their siblings but without statistical significance. It is consequently evident that decreased antioxidative protection and simultaneous free radical (FR) overproduction occur in diabetic children and that there is a similar, but not significant, tendency in their siblings. The findings warrant reducing OS in diabetic children and postponing disease onset in susceptible relatives.