Elevated Levels of Toxic Bile Acids in Serum of Cystic Fibrosis Patients with CFTR Mutations Causing Pancreatic Insufficiency.

Hepatobiliary involvement is a hallmark in cystic fibrosis (CF), as the causative CF Transmembrane Conductance Regulator (CFTR) defect is expressed in the biliary tree. However, bile acid (BA) compositions in regard to pancreatic insufficiency, which is present at an early stage in about 85% of CF patients, have not been satisfactorily understood. We assess the pattern of serum BAs in people with CF (pwCF) without CFTR modulator therapy in regard to pancreatic insufficiency and the CFTR genotype. In 47 pwCF, 10 free and 12 taurine- and glycine-conjugated BAs in serum were prospectively assessed. Findings were related to genotype, pancreatic insufficiency prevalence (PIP)-score, and hepatic involvement indicated by serum liver enzymes, as well as clinical and ultrasound criteria for CF-related liver disease. Serum concentrations of total primary BAs and free cholic acid (CA) were significantly higher in pwCF with higher PIP-scores (p = 0.025, p = 0.009, respectively). Higher total BAs were seen in pwCF with PIP-scores ≥0.88 (p = 0.033) and with pancreatic insufficiency (p = 0.034). Free CA was higher in patients with CF-related liver involvement without cirrhosis, compared to pwCF without liver disease (2.3-fold, p = 0.036). pwCF with severe CFTR genotypes, as assessed by the PIP-score, reveals more toxic BA compositions in serum. Subsequent studies assessing changes in BA homeostasis during new highly effective CFTR-modulating therapies are of high interest.

Cross-sectional analysis of universal vitamin D supplementation in former East Germany during the first year of life.

BACKGROUND: Universal vitamin D supplementation is controversial. Preventative examinations and public health initiatives in former East Germany that included vitamin D prophylaxis for children were regulated by official recommendations and guidelines. The aim of this study is to analyse the impact of a standardised nationwide guideline for universal supplementation with 400 International Units (IU) vitamin D3/day during the first year of life on clinical and biochemical parameters and the influence of surrounding factors. METHODS: This is a cross-sectional analysis looking at data from a field study of 3481 term-born children during their first year of life that was conducted in 1989. RESULTS: There were no significant clinical signs of rickets. 25 hydroxyvitamin D (25(OH)D) (mean and SEM, total analyses n=572) after birth (n=28) was 36(7) nmol/L, at 1 month 64(4) nmol/L (n=70, p<0.0001), 91(5) nmol/L at 3 months (n=95, p<0.0001), 65(8) nmol/L at 8 months (n=21, p=0.005) and ranged between 33 and 109 nmol/L until 12 months. Less than 0.2% of analyses revealed pathological levels for calcium or phosphate. Alkaline phosphatase (ALP) levels (n=690) were >1500 U/L (95th percentile) in 3.6%. Participants were on breastmilk or vitamin D-free formula, with solids added from 6 months of age. There were seasonal variations in 25(OH)D levels with a rise during spring and autumn. Thus this analysis is unique as sun exposure and supplementation can be considered as the only vitamin D sources. CONCLUSIONS: We conclude that universal supplementation with 400 IU of vitamin D3 during the first year of life is safe and provides sufficient 25(OH)D levels in Germany.

Expression of components of the IGF axis in childhood acute myelogenous leukemia.

BACKGROUND: Insulin-like growth factor (IGF) system as regulator for cellular proliferation is of particular interest in search for new prognostic approaches in cancer treatment. PROCEDURE: We analyzed the mRNA expression profile of IGF-I, -II, and IGFBP-2, -3 in 50 children with previously untreated AML (mean age 10.8 +/- 4.8 years; patients in CCR n = 20, patients with relapse during later course of disease n = 15). MNC samples from peripheral blood as well as bone marrow of healthy donors were used as controls. RESULTS: IGFBP-2 expression was significantly higher in AML cells than in healthy cells of peripheral MNC (P < 0.001) and of bone marrow cells (P < 0.01). Conversely, AML cells showed significantly lower IGFBP-3 and IGF-I gene expression compared to controls (P = 0.02; P < 0.001). Patients with relapse (median +/- range: 0.0929 +/- 0.049) during later course of disease demonstrated higher IGFBP-2 expression compared to patients in CCR (0.0121 +/- 0.047; P = 0.06) at time of diagnosis. A multivariate analysis identified the IGFBP-2 mRNA expression as an independent factor for the prediction of relapse. Furthermore, the probability of relapse-free survival (RFS) in patients with IGFBP-2 mRNA level >0.1000 was 28%; whereas, the probability of RFS in patients with IGFBP-2 mRNA level <0.1000 was 62% (P = 0.04, log-rank test). No prognostic influence could be found for the other investigated genes. CONCLUSIONS: Results identified different expressions of IGF components between normal and AML cells. Patients with IGFBP-2 mRNA levels up to 0.1000 (relative to KG1 cell line) more likely developed a relapse. Identification of these patients at diagnosis may allow more individualized treatment.

Effect of dexamethasone on the IGFBP-1 regulation in premature infants during the first weeks of life.

BACKGROUND: Early glucosteroid treatment in preterm infants has a negative impact on physical growth and development. So far, data on dexamethasone effect on the GH/IGF axis and the clinical outcome are conflicting. OBJECTIVE: Therefore, we studied the effect of dexamethasone treatment on parameters of somatic growth and on the secretion of insulin like growth factors (IGFs) and insulin like growth factors binding proteins (IGFBPs) in preterm infants. METHODS: In 75 preterm infants somatic development was assessed at birth and after 3 months of corrected age. IGF-I/II and IGFBP-1-3 were measured at the same time. According to their treatment regime the infants were assigned to the dexamethasone treated or the non-treated group. RESULTS: At 3 months the 13 infants with dexamethasone had a lower body weight, slightly lower body length and a lower head circumference. IGF-II (464.4 +/- 97.4 vs 638 +/- 201.4 mug/l, p = 0.001) and IGFBP-3 (1800 +/- 426 vs 2105 +/- 547 mug/l, p = 0.045) were significantly reduced under the influence of glucocorticoids, whereas IGFBP-1 was elevated (59.6 +/- 61.0 vs 21.1 +/- 21.7 mug/l, p = 0.002). The ratio IGFBP-3/(IGFBP-1 + 2) was reduced in the dexamethasone group (1.827 +/- 0.868 vs 3.098 +/- 1.898 mug/l, p = 0.016), implying a significant retardation in the somatic development. CONCLUSION: Dexamethasone impairs IGF and IGFBP secretion and stimulates IGFBP-1, an inhibitor of IGF-I. These pathways might contribute to alterations of the GH/IGF axis, particularly the ratio IGFBP-3/(IGFBP-1 + 2).

Associations between ghrelin levels in serum of preterm infants and enteral nutritional state during the first 6 months after birth.

BACKGROUND: Previous studies have suggested a possible influence of ghrelin on foetal growth. After birth, the regulation of this newly discovered orexigenic peptide is largely unknown. OBJECTIVE: To study the relationship between circulating levels of ghrelin and enteral nutritional state in preterm infants during the first months of life. METHODS: Ghrelin levels were measured in a cross-sectional study by radioimmunoassay on the second day after birth (n = 51), at 3 months (n = 63) and at 6 months (n = 53) of corrected postnatal age. Complete data sets of auxological parameters, biochemical values, perinatal diseases, nutritional management and therapy were determined. RESULTS: All infants showed levels of ghrelin in postnatal serum. In correlation analyses and multivariable linear regression models, ghrelin was strongly related to enteral caloric intake on the second day after birth (all P < 0.01). At 6 months of corrected postnatal age, infants who were exclusively breastfed/formula fed had significantly lower ghrelin levels than infants with solid foods (Mann-Whitney U-test: P = 0.027). CONCLUSION: Ghrelin levels were positively correlated with the enteral nutritional state in preterm infants on the second day after birth. The introduction of solid foods increased the ghrelin levels in a group of preterm infants at 6 months of corrected postnatal age.

Associations of the IGF/IGFBP axis and respiratory diseases in neonatal patients during the first 6 months of life.

OBJECTIVES: To analyse IGFs in respect to somatic growth and neonatal diseases during the first 6 months of life. METHODS: IGF-I, IGF-II and IGF binding proteins (IGFBP-1, -2, -3) were determined by immunoassays in neonatal patients after birth (n = 67) and at 3 (3 mo; n = 75) and 6 months (6 mo; n = 47) of corrected postnatal age. Data on growth were tested for associations to the gestational age, birth weight (bw) SDS (/-2 SDS), neonatal morbidity and therapeutic strategies. RESULTS: All IGFs and IGFBPs changed significantly between birth and 3 mo of corrected age (p < 0.05). Perinatal respiratory diseases influenced IGF-II at 3 mo, and bronchopulmonary dysplasia IGF-II at 3 and 6 mo (all p < 0.05). IGF-I differed between the subgroups bw /-2 SDS (p < 0.05). At 3 mo, IGFBP-1 was significantly increased in infants with glucocorticoid administration during the first four weeks of life. CONCLUSION: The first months of life are characterised by a pole reversal of the somatotropic axis: IGFBP-1 and -2 decrease and IGFBP-3 increases. Respiratory diseases with an origin in the neonatal period, glucocorticoid therapy and low birth weight have an impact on the IGF pattern up to 6 mo. Prospective studies are necessary to investigate, whether the described link between the IGF/IGFBP axis and respiratory morbidity in neonatal patients has an impact on development in later infancy.

The potential of digital X-ray radiogrammetry (DXR) in the assessment of osteopenia in children with chronic inflammatory bowel disease.

BACKGROUND: Loss of bone mass is a known complication of chronic inflammatory bowel disease (IBD) in children. The gold standard in the evaluation of bone mineral density (BMD) is dual energy X-ray absorptiometry (DXA). OBJECTIVE: In this preliminary study we evaluated digital X-ray radiogrammetry (DXR) which estimates BMD (DXR-BMD) from hand radiographs in children with IBD. MATERIALS AND METHODS: A total of 26 children with IBD (10 girls, 16 boys; age range 10-18 years) underwent DXR for the calculation of DXR-BMD and metacarpal index (DXR-MCI) using the Pronosco X-posure system. The results were compared with a local reference database and correlated with the results of DXA. RESULTS: DXR-BMD was 0.36-0.56 g/cm(2) (median 0.46 g/cm(2)) in Crohn disease patients and 0.38-0.63 g/cm(2) (median 0.48 g/cm(2)) in ulcerative colitis patients. DXR-MCI was 0.29-0.49 in Crohn disease patients and 0.28-0.53 in ulcerative colitis patients. The Z-scores were reduced to <-1 SD in five Crohn disease patients and in six ulcerative colitis patients. The coefficients (r) for the correlations between DXR-BMD and DXA-BMD were 0.78 for the lumbar spine and 0.61 for the proximal femur (P<0.01), and between DXR-MCI and DXA-BMD were 0.78 for the lumbar spine and 0.51 for the proximal femur (P<0.01). CONCLUSIONS: DXR seems to be able to estimate cortical osteopenia in children with chronic IBD. The DXR results showed a positive correlation with DXA results.

Does insulin-like growth factor 1 contribute in red blood cell transfusions to the pathogenesis of retinopathy of prematurity during retinal neovascularization?

BACKGROUND: Red blood cell (RBC) transfusions are associated with the development of retinopathy of prematurity (ROP). During the period of retinal neovascularization a rise of insulin-like growth factor 1 (IGF-1) may trigger rapid growth of new blood vessels. OBJECTIVES: To study endocrine factors in RBC transfusions that might be of importance for ROP. METHODS: IGF-1, IGF-2 and their binding proteins 1-3 (IGFBP-1-3) were determined by radioimmunoassays in 7 very-low-birthweight (VLBW) infants with ROP >or= stage 2 receiving a RBC transfusion, in 10 controls (VLBW infants with ROP

Assessment of skeletal age at the wrist in children with a new ultrasound device.

BACKGROUND: Determination of skeletal development in children is important. The most common method of evaluation uses the standards of Greulich and Pyle (G and P) to assess the left hand radiograph. Numerous assessments may be made during follow-up. OBJECTIVE: The aim of our study was to compare the accuracy of a new sonographic method with the standard radiographic method. MATERIALS AND METHODS: Seventy consecutive patients (age 6-17 years; 34 girls, 36 boys) underwent radiography of the left hand, followed by sonographic examination of the same hand using the BonAge system (Sunlight Medical Ltd., Israel). This system evaluates the relationship between the velocity of sound passing thorough the distal radial and ulna epiphysis and growth, using gender- and ethnicity-based algorithms. One experienced paediatric radiologist analysed the radiograph and assigned bone age scores based on the G and P atlas for the whole left hand and for the distal radius alone. The radiologist was blinded to the chronological age (CA), height of the patient and the BonAge result. Correlation between BonAge and G and P was undertaken. RESULTS: In 65 patients, BonAge measurement could be performed successfully. In five patients, the scanning process was impossible using the ultrasound device. The r(2) (r is the Pearson correlation coefficient) of the BonAge ultrasound measurement and the G and P method was 0.82. The averaged accuracy (i.e. absolute difference in years between G and P reading and BonAge ultrasonic results) was calculated. Results were similar for boys and girls: 1.0+/-0.8 years for the whole left hand and 0.8+/-0.7 year for the distal radius. On average, the difference between BonAge and CA is the same as the difference between G and P and CA, i.e. 1.4 years. CONCLUSIONS: The BonAge device demonstrates the ability of ultrasound to produce an accurate assessment of bone age. The results are highly correlated with skeletal age evaluated conventionally using the G and P method. Obvious advantages of the ultrasound device are objectivity, lack of ionizing radiation, and easy accessibility.

Conjugated linoleic acid modulation of cell membrane in leukemia cells.

This study compared the cellular uptake of pure conjugated linoleic acid isomers (CLA(9c,11t) and CLA(9c,11c)) to linoleic acid (LA) and their effects on polyunsaturated fatty acid (PUFA) synthesis, its metabolism into conjugated long chain fatty acids (FAs) by desaturation and chain-elongation as well as cell proliferation and the associated anticarcinogenic effects on various human leukemia cell lines (K562, REH, CCRF-CEM and U937 cells). Furthermore, selective effects of this individual isomers of CLA on desaturation steps involved in the biosynthesis of PUFAs associated with cell growth were investigated. CLA isomers supplemented in the culture medium was readily incorporated and esterified into phospholipids (PLs) in the four cell lines in a concentration- and time-dependent manner. The incorporation of the specific CLA isomers in PLs was similar to LA. All four incubating leukemia cells (40 microM CLA for 48 h) showed very high cellular CLA content in PLs (range: 32-63 g FA/100 g total phospholipid fatty acid) affected by the nature of CLA and the cell type. Supplementation with CLA or LA altered also cell membrane composition by n-6 PUFA synthesis. Accordingly, CLA metabolism interferes with LA metabolism. We were able to show that CLA isomers are converted by the leukemia cells of the same metabolic pathway into conjugated diene fatty acids (CDFAs) as LA into non-conjugated PUFAs. In this view, the gas chromatography-flame ionization detector detection of major CDFAs (CD-18:3, CD-20:2 and CD-20:3) in cell membrane of CLA-treated cultures resulted from successive Delta6-desaturation, elongation and Delta5-desaturation of CLA isomers. However, in comparison to LA, relatively lower amounts of elongation and/or desaturation metabolites were detected for CLA(9c,11t), and only minor amounts or trace CDFAs were observed for CLA(9c,11c). Furthermore, CLA(9c,11t) revealed only very low levels of CD-20:4 FA and no CLA(9c,11c)-conversion could be detected. The metabolization of CLA indicated that CLA(9c,11c)60 microM) had the CLA type dependent antiproliferative effects. Thus, the 9cis,11trans- and the 9cis,11cis-CLA isomers regulate cell growth and survival in different leukemia cell types through their existence alone and/or by their inhibitory effects of desaturase activity.

Thyroid hormone response to moderate hypothermia in severe brain injury.

OBJECTIVE: To examine the effect of controlled moderate hypothermia on thyroid response in head-injured patients. DESIGN: Prospective, controlled, randomized study. SETTING: University hospital intensive care unit (ICU). PATIENTS: Twenty-eight patients with severe blunt head injury (Glasgow Coma Scale < or =9). INTERVENTION: Patients were randomly assigned to a hypothermia or a normothermia group. Hypothermia (32-33 degrees C) was induced within 8 h after trauma and maintained for a mean of 36 h. All patients were sedated and mechanically ventilated. MEASUREMENTS AND RESULTS: Thyroid-stimulating hormone( TSH), free and total triiodothyronine (FT3/TT3), reverse triiodothyronine (RT3) and thyroxine (FT4/TT4) were measured during the hypothermia or corresponding normothermia period, after regaining normothermia and 4-6 days later. Of 28 patients included in the study, 11 subjects were treated with hypothermia and 13 patients with normothermia. Four patients had to be excluded. In both groups, serum concentrations of TT3 and FT3 were just below the lower normal range whereas RT3 serum concentrations were near the upper limit of the normal range. TSH serum concentrations were not increased. No statistically significant intra- or inter-group differences were observed. CONCLUSIONS: Thyroid hormone patterns during moderate hypothermia in head-injured patients did not differ from the well known "low T3 state" which is observed in other forms of severe illness.

Advanced glycation end products in children with chronic renal failure and type 1 diabetes.

Serum levels of advanced glycation end products (AGEs) are markedly elevated in adults with chronic renal failure (CRF) and diabetes mellitus. Accumulation of AGEs in tissues contributes to the development of long-term complications. Up to now little has been known about the formation of AGEs in childhood. We determined serum levels of the well known AGEs pentosidine and Nvarepsilon-carboxymethyllysine (CML) in children with CRF (n=12), end-stage renal disease (ESRD) (n=9), renal transplantation (n=12), and type 1 diabetes mellitus (n=42) and in healthy children (n=20). Pentosidine was measured by high-performance liquid chromatography (HPLC), CML by a competitive enzyme-linked immunosorbent assay (ELISA) system. Serum levels of pentosidine and CML were significantly higher in the children with CRF and ESRD than in controls (P< 0.001), but nearly within the normal range after transplantation. Both AGEs showed a significant negative correlation with creatinine clearance (P< 0.001). During a single session of low-flux hemodialysis, total pentosidine and CML levels did not change. Free pentosidine, however, was reduced by 78% (P=0.04). Diabetic children showed significantly elevated pentosidine levels (P< 0.001) despite normal renal function. We conclude that, similar to adults, increased formation and accumulation of AGEs also exist in children with CRF and type 1 diabetes mellitus. At present the best prevention of AGE-related complications is an early renal transplantation in children with ESRD, as well as a careful metabolic monitoring of diabetics.