High-dose Vitamin D Supplementation in Multiple Sclerosis - Results From the Randomized EVIDIMS (Efficacy of Vitamin D Supplementation in Multiple Sclerosis) Trial

Background: Epidemiological, preclinical, and non-interventional studies link vitamin D (VD) serum levels and disease activity in multiple sclerosis (MS). It is unclear whether high-dose VD supplementation can be used as an intervention to reduce disease activity. Objectives: The study aimed to compare the effects of every other day high- (20,400 IU) versus low-dose (400 IU) cholecalciferol supplementation on clinical and imaging markers of disease activity in patients with relapsing-remitting MS or clinically isolated syndrome. Methods: The EVIDIMS (efficacy of vitamin D supplementation in multiple sclerosis) trial was a multicentre randomized/stratified actively controlled explorative phase 2a pilot trial with a double-blind intervention period of 18 months, add on to interferon-β1b. Results: Fifty-three patients were randomized, and 41 patients completed the study. Cholecalciferol supplementation was well tolerated and safe in both arms. After 18 months, clinical (relapse rates, disability progression) and radiographical (T2-weighted lesion development, contrast-enhancing lesion development, brain atrophy) did not differ between both treatment arms. Post-study power calculations suggested that the sample size was too low to prove the hypothesis. Conclusions: The results neither support nor disprove a therapeutic benefit of high-dose VD supplementation but provide a basis for sound sample size estimations in future confirmatory studies. www.clinicaltrials.gov/NCT01440062

Early morbidity and dose–volume effects in definitive radiochemotherapy for locally advanced cervical cancer: a prospective cohort study covering modern treatment techniques

PURPOSE: Predicting morbidity for patients with locally advanced cervix cancer after external beam radiotherapy (EBRT) based on dose–volume parameters remains an unresolved issue in definitive radiochemotherapy. The aim of this prospective study was to correlate patient characteristics and dose–volume parameters to various early morbidity endpoints for different EBRT techniques, including volumetric modulated arc therapy (VMAT) and adaptive radiotherapy (ART). METHODS AND MATERIALS: The study population consisted of 48 patients diagnosed with locally advanced cervix cancer, treated with definitive radiochemotherapy including image-guided adaptive brachytherapy (IGABT). Multiple questionnaires (CTCAE 4.03, QLQ-C30 and EORTC QLQ-CX24) were assessed prospectively for patients treated with different EBRT techniques, including online adaptive VMAT. Contouring and treatment planning was based on the EMBRACE protocols. Acute toxicity, classified as general, gastrointestinal (GI) or genitourinary (GU) and their corresponding dose–volume histograms (DVHs) were first correlated by applying least absolute shrinkage and selection operator (LASSO) and subsequently evaluated by multiple logistic binomial regression. RESULTS: The treated EBRT volumes varied for the different techniques with ~2500 cm(3) for 3D conformal radiotherapy (3D-CRT), ~2000 cm(3) for EMBRACE‑I VMAT, and ~1800 cm(3) for EMBRACE-II VMAT and ART. In general, a worsening of symptoms during the first 5 treatment weeks and recovery afterwards was observed. Dose–volume parameters significantly correlating with stool urgency, rectal and urinary incontinence were as follows: bowel V(40Gy) < 250 cm(3), rectum V(40Gy) < 80% and bladder V(40Gy) < 80–90%. CONCLUSION: This prospective study demonstrated the impact of EBRT treatment techniques in combination with chemotherapy on early morbidity. Dose–volume effects for dysuria, urinary incontinence, stool urgency, diarrhea, rectal bleeding, rectal incontinence and weight loss were found

Dynamic adaptation of myocardial proteome during heart failure development

<div><p>Heart failure (HF) development is characterized by huge structural changes that are crucial for disease progression. Analysis of time dependent global proteomic adaptations during HF progression offers the potential to gain deeper insights in the disease development and identify new biomarker candidates. Therefore, hearts of TAC (transverse aortic constriction) and sham mice were examined by cardiac MRI on either day 4, 14, 21, 28, 42, and 56 after surgery (n = 6 per group/time point). At each time point, proteomes of the left (LV) and right ventricles (RV) of TAC and sham mice were analyzed by mass spectrometry (MS). In TAC mice, systolic LV heart function worsened from day 4 to day 14, remained on a stable level from day 14 to day 42, and showed a further pronounced decline at day 56. MS analysis identified in the LV 330 and in RV 246 proteins with altered abundance over time (TAC vs. sham, fc≥±2). Functional categorization of proteins disclosed the time-dependent alteration of different pathways. Heat shock protein beta-7 (HSPB7) displayed differences in abundance in tissue and serum at an early stage of HF. This study not only provides an overview of the time dependent molecular alterations during transition to HF, but also identified HSPB7 as a novel blood biomarker candidate for the onset of cardiac remodeling.</p></div

IPA based biological function activity analysis.

<p>For analysis all proteins of LV which showed differential abundance (TAC vs. sham, fc≥2) only in the time period between day 14 and 42 after TAC were used. A positive z-score predicts activation (orange), a negative score (purple) predicts inhibition of the biological process.</p

Heat map of abundance changes of proteins associated with cardiovascular diseases.

<p>Ratios (TAC/sham) of proteins in left (LV) and right ventricle (RV) with a ≥2 fold change on at least one examined time point. The changes of common HF markers like ANF (ANP), MYH7 (β-MHC) and ACTS (α-actin) were markedly higher in LV than in RV especially at the early time points. After 56 days in both ventricles comparable alterations regarding these proteins were observed. ECM (extracellular membrane), n.d. (not detected) * Labeling as exported from Rosetta Elucidator^® package.</p

Heat map of canonical pathways affected during disease progression in LV and RV.

<p>Canonical pathways with a significant enrichment of altered proteins (TAC vs. sham; fc≥2) are displayed for each examined time point for LV and RV. Significance values were assessed using Fisher’s exact test and are shown for LV from light blue (p<0.05) to dark blue (p<0.001) and for RV from light green (p<0.05) to dark green (p<0.001).</p

TAC-induced changes in protein abundance.

<p>The number of proteins with an at least 2-fold change in abundance TAC vs. sham are shown. (A<b>)</b> Over time, 330 different proteins in the left ventricle (LV) and 246 proteins in the right ventricle (RV) were altered after TAC, of which 82 displayed alterations in both ventricles. <b>(</b>B<b>)</b> The number of altered proteins per time point. The strongest changes in protein pattern in LV were observed in the early phase after TAC at day 14 and 21, whereas strongest differences in RV were seen at day 28, 42, and 56. Especially cytoplasmic and mitochondrial proteins were affected.</p

Abundances of small heat shock proteins.

<p>(A) Intensities (normalized to total protein) of tissue HSPB6 detected by Western Blot analysis on individual sample level (n = 6 per group/time point). Over all time points HSPB6 abundances are significantly higher in TAC mice compared to sham mice (two-way ANOVA, Bonferroni post-test, *** p<0.001; ** p<0.01). (B) Intensities (normalized to total protein) of HSPB7 detected by western Blot analysis on individual sample level (n = 6 per group/time point). Already at day 4 HSPB7 could be found in significantly higher levels in the heart tissue of TAC mice when compared to sham (two-way ANOVA, Bonferroni post-test, *** p<0.001). The high expression remained over all time points and is comparable to the proteome analysis. (C) Serum concentrations of HSPB7 analyzed by ELISA on individual sample level (n = 5–6 per group/time point) showed a significant (p<0.001, two-way ANOVA, Bonferroni post-test) increase in TAC mice after 4 days of pressure overload. Higher amounts of protein could be found also at all other time points in TAC mice, whereas HSPB7 was not detectable in the sham group.</p