Computational protocol: Docosahexanoic Acid Plus Vitamin D Treatment Improves Features of NAFLD in Children with Serum Vitamin D Deficiency: Results from a Single Centre Trial

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Protocol publication

[…] The assessment of fibrillar collagen deposition within the liver biopsy was evaluated in Sirius Red (SR) stains, as previously [,]. Briefly, SR stained slides were scanned by a digital scanner (Aperio Scanscope CS System, Aperio Technologies, Inc, Oxford, UK) and processed by ImageScope. An image analysis algorithm has been used to quantify the proportion of SR-stained area. The algorithm was applied on the entire section (Part A and B of ). The extent of collagen deposition was expressed as the proportion (%) of SR-stained area with respect to the total biopsy area, providing a quantitative value on a continuous scale. Only biopsies containing at least 5 portal tracts were considered.In order to establish reference values for fibrillar collagen in normal liver samples, specimens from 6 lean, non-diabetic children (boys, 4; girls, 2; median age: 13 years, range, 12–16 years) without liver disease were used as controls, as previously []. These fragments were obtained from patients who underwent laparotomy or laparoscopic procedures (for cholecystectomy), from liver donors (orthotopic liver transplantation) or incidental “normal” liver biopsies (children exhibiting persistent/intermittent elevations of liver enzymes for >6 months). Informed consent in writing was obtained from next of kin, caretakers, or guardians on behalf of the children enrolled in this study [,]. [...] The data were analyzed using a STATISTICA (version 2010, Chicago, IL, USA). Continuous variables were expressed as mean ± standard deviation (SD). Data distribution was checked for normality by the Kolmogorov-Smirnov test. Data were analyzed using the intention-to-treat principle and the values recorded at baseline were compared to values recorded at 6 and 12 months in all patients, regardless of treatment duration. Baseline and follow up characteristics were tested for differences by Student’s t-test (p<0.05). The change of anthropometrical and laboratory values, between placebo and treatment groups, was evaluated using analysis of variance (ANOVA) with repeated measures. Difference between proportions were tested using the Chi-square test. Univariate correlations were investigated with Pearson’s correlation. Multivariable logistic regression analysis was used to test the independence of associations between end of study vitamin D concentrations as the key exposure and histological characteristics after adjusting for BMI, change in BMI between baseline and follow up and basal values of Vitamin D. […]

Pipeline specifications

Software tools ImageScope, Statistica
Applications Miscellaneous, Whole slide imaging
Organisms Homo sapiens
Chemicals Triglycerides, Vitamin D