Absolute deviation in median BMI-for-chronological-age and BMI-for-height-age between samples with tall or short stature from the sample with normal stature.

<p>A smaller deviation indicates that the median BMI of short or tall sample is more similar to the median BMI of the sample with normal stature. For boys with a short stature in the ages 4 up to 14 years of age, expressing BMI according to height-age resulted in a smaller deviation from the BMI distribution of children with a normal stature than when expressing BMI to chronological age, whereas in boys with a tall stature this was true in the ages 4-16 years. In girls, expressing BMI to height-age resulted in a smaller deviation in the ages 2-10 years if of being short stature and in the ages 2-17 years when being tall.</p

Massive parallel sequencing in steroid-resistant nephrotic syndrome (SRNS)

<p><strong>ABSTRACT</strong></p> <p><strong>Introduction</strong>. To date abnormalities in more than 20 genes have been associated with SRNS. Sequencing of all SRNS genes requires ~ 600 PCR amplicons, rendering conventional mutation testing unfeasible due to financial and time constraints. Hence, current screening algorithms usually include only the most common disease genes and/or use preselection according to additional phenotypic criteria. This practice typically allows for mutation detection in ~15% of patients. We have evaluated targeted NGS screening of SRNS patients enrolled in the PodoNet registry who were found negative for mutations in the first-line SRNS-associated genes.</p> <p><strong>Material and Methods</strong>. Molecular analysis of 31 known or plausible SDNS disease genes was performed by NGS using a custom-designed multiplex PCR kit (MASTR FSGS, Multiplicom). The pilot group consisted of 22 patients with 4.7 years median age at disease onset (range 0.5-20 years), positive family history in 68%, parental consanguinity in 18% and chronic renal insufficiency at last observation in 59%.</p> <p><strong>Results.</strong> Mean coverage was 1269x (median 1286, range 929-1826). 18/22 runs had at least 15x read depth covering 99% of the target sequences. One patient was diagnosed with hereditary SRNS due to a previousy described homozygous pathogenic mutation in SMARCAL1 gene. In addition, three novel sequence variants in the genes PLCE1 (homozygous), LAMB2 (homozygous) and WT1 (heterozygous) were detected. In silico studies support their classification as pathogenic, even though the patients do not present the characteristic clinical and/or histopathological features typically reported for patients with mutations in these genes.</p> <p><strong>Conclusions</strong>. Our detection of pathogenic mutations in 4 out of 22 SRNS patients screened negative by conventional selective screening approaches support targeted NGS testing in all SRNS patients, regardless of age at diagnosis, absence of extrarenal manifestations or histological subtype. We anticipate that systematic NGS screening of the SRNS cohorts collected in EURenOmics will allow re-evaluation of mutation incidence rates in SRNS and become the new standard of genetic diagnostics in this condition.</p

Digital scanning and diagnostic scoring of kidney biopsies from children with steroid resistant nephritic syndrome

<p><strong>Background:</strong> Digital pathology is an attractive new tool for multicenter research. Practical improvements on conventional pathology include parallel evaluation by several experts in distant locations and reproducible identification of individual lesions. Also, digital image analysis allows more objective measures of cell staining and 3D reconstruction of glomeruli. </p> <p> </p> <p><strong>Design:</strong> We are establishing a digital pathology archive within WP2 of EURenOmics. Kidney biopsies from children with steroid resistant nephrotic syndrome (SRNS) enrolled in the PodoNet registry are collected. Analysis will be harmonized with digital pathology scoring developed by the NEPTUNE study (Nephrotic Syndrome Study Network) in the US.</p> <p> </p> <p><strong>Methods:</strong> The material is scanned at high resolution in Heidelberg at the Hamamatsu Tissue Image and AnalysisCenter using a Hamamatsu Nanozoomer. Digital images of existing electron microscopy and immunofluorescence scans and original pathology reports are also collected. After anonymization and manual quality controls (e.g. checking correct focus in all areas) data is stored centrally for remote review.</p> <p>Digital images are first annotated, i.e. each glomerulus is given a number which can be tracked across different section levels. Histopathological scoring of each glomerulus in each section level will then be performed in an unbiased fashion by independent blinded pathologists. These scores will be correlated to clinical and genetic information, conventional histopathological diagnoses and molecular profiles obtained by multi-omics profiling. </p> <p> </p> <p><strong>Progress:</strong> So far 83 kidney biopsies have been collected and at least another 180 are expected. Test runs of different stain types have produced good quality images. Bulk scanning and annotations are commencing, so that pathology reviews will start in spring 2014. </p> <p> </p> <p><strong>Outlook:</strong> The new digital pathology archive for children with SRNS will enable standardized review and objective scoring of histopathological findings. We hope that this will improve correlations of histological findings with clinical outcomes and biomarkers. Setting up the relevant infrastructure will allow extension of the project to other registries within WP2.</p

Bland Altman plot comparing the composite measure of intima thickness and medial thickness (IT + MT) by ultra-high-frequency ultrasound with conventional cIMT.

<p>Bland Altman plot comparing the composite measure of intima thickness and medial thickness (IT + MT) by ultra-high-frequency ultrasound with conventional cIMT.</p

Mean changes in systolic and diastolic blood pressure according to pre- and perinatal variables, independently of current child’s BMI z-score.

<p>The values presented are β regression coefficients and 95% confidence intervals (shown in parenthesis). Model 1 is adjusted to each variable in the table and additionally for age (in months). Model 2 is similar to Model 1 but additionally adjusted for child’s BMI z-score.</p><p>β, regression coefficient; 95%CI, 95% confidence interval; BP, blood pressure.</p><p>Mean changes in systolic and diastolic blood pressure according to pre- and perinatal variables, independently of current child’s BMI z-score.</p

Hemodiafiltration is associated with reduced inflammation, oxidative stress and improved endothelial risk profile compared to high-flux hemodialysis in children

<div><p>Randomized trials in adults have shown reduced all-cause and cardiovascular mortality on hemodiafiltration (HDF) compared to high-flux hemodialysis (HD), but the mechanisms leading to improved outcomes are not clear. We studied biomarkers of inflammation, oxidative stress, anti-oxidant capacity and endothelial dysfunction in 22 children (13 female, age 8–15 years). All children received HD for at least 3 months, and were then switched to HDF, keeping all dialysis related parameters and dialysis time constant. All the biomarkers of inflammation (ß2-microglobulin, IL-6, IL-10, high sensitive C-reactive protein [hsCRP]), oxidative stress (nitrotyrosine, advanced glycation end-products [AGEs], oxidized low density lipoprotein [ox-LDL] and anti-oxidant capacity) and endothelial dysfunction (asymmetric dimethyl arginine [ADMA], symmetric dimethyl arginine [SDMA]), were comparable between incident and prevalent patients on HD, suggesting that even a short dialysis vintage of 3 months on HD increases inflammation and endothelial stress. After 3 months of HDF therapy there was a significant reduction in ß2-microglobulin (p<0.001), hCRP, ADMA, SDMA, AGEs, ox-LDL (p<0.01 for all) and an increase in total antioxidant capacity (p<0.001) compared to HD. All children were maintained on the same dialyser, dialysis water quality, dialysis time and blood flow speeds suggesting that improved clearances on HDF led to an improved biomarker profile. Even in children with residual renal function there was a significant reduction in ß2 microglobulin, hsCRP, SDMA, ox-LDL and AGEs on HDF compared to HD. Children with a lower blood flow had higher inflammatory status (higher IL-6/IL-10 ratio; p = 0.04, r = -0.43). Children who achieved a higher convective volume (≥median 12.8L/m²) had lower ox-LDL (p = 0.02). In conclusion, we have shown that a significant improvement in inflammation, antioxidant capacity and endothelial risk profile is achieved even within a short time (3 months) on HDF compared to HD treatment.</p><p><b><i>Trial Registration</i>:</b> ClinicalTrials.gov: <a href="https://clinicaltrials.gov/ct2/show/NCT02063776" target="_blank">NCT02063776</a>.</p></div

Changes in ultra-high-frequency ultrasound measures at one year follow-up.

<p><b>(Fig 4A</b>—Comparison of baseline and 1-year follow-up measures of common carotid artery (CCA) medial thickness (MT) in transplanted children, <b>Fig 4B</b>—Comparison of baseline and 1-year follow-up measures of dorsal pedal artery intimal thickness (IT) in transplanted children).</p

Correlation of ultra-high-frequency ultrasound measures with biochemical markers.

<p><b>(Fig 3A–</b>Association of serum phosphate with common carotid artery (CCA) medial thickness (MT), <b>Fig 3B–</b>Association of serum parathyroid hormone level (log transformed) with CCA MT, <b>Fig 3C–</b>Association of mean arterial pressure standard deviation score (SDS) with CCA MT, <b>Fig 3D–</b>Association of diastolic BP SDS with dorsal pedal artery MT).</p

Ultra-high-frequency ultrasound measurements of intima thickness (IT), medial thickness (MT) and the composite measure of IMT in carotid (Fig 2A) and dorsal pedal (Fig 2B) arteries in healthy controls and children with CKD4-5 and on dialysis (CKD4-5D) and transplanted patients.

<p>CCA–common carotid artery.</p

Hemodiafiltration is associated with reduced inflammation, oxidative stress and improved endothelial risk profile compared to high-flux hemodialysis in children - Fig 3

<p>Percent increase in TAC and percent decrease in SDMA from HD to HDF was more pronounced in patients with higher blood flow rate (A,B). Higher blood flow rate was also associated with lower IL-6 to IL-10 ratio on HDF (C). Percent change = [(HDF-HD)/HD]*100.</p