Furthermore, sections were stained with antibodies against myosin slow and fast (Novocastra, Milton Keynes, UK, 1:100), �� sarcoglykan (Novocastra, 1:200), and N-terminal utrophin (Novocastra, 1:200). Thick sections of 4 ��m were cut of the resin embedded tissue and stained with toluidine selleck chemical blue.The quantification of myopathy was based on characteristic features of acute myopathy in intensive care, namely type II fibre atrophy (numerous scattered angular, atrophic fibers identified as type 2 fibers by ATPase and myosin stains), muscle necrosis, and selective loss of myosin filaments and scored as follows [4,14,19]: no signs of myopathy (score = 0), signs of mild myopathy (score = 1), signs of moderate myopathy (score = 2), signs of severe myopathy (score = 3), and signs of very severe myopathy (score = 4).

Hence, the CIPNM severity sum score consisting of the CIP and CIM scores determined on days 0 and 14 ranged from 0 (no CIM, no CIP) to 8 (very severe CIP, very severe CIM).Secondary outcomes were to assess the effect of early IVIG versus placebo on mortality from any cause within a 28-day period and length of the ICU stay. Furthermore, we investigated the course of CIPNM from baseline to Day 14 in all patients.Sample sizeThe software PASS 11 (NCSS, Kaysville, UT, USA) was used for sample size calculation. Group sample sizes of 2 �� 30 patients achieve 81% power to detect a difference in CIPNM sum score of 1.5 between the intervention group (estimated score of 4.0) and the control group (estimated score of 2.5) given standard deviations of 2.

0 and at a two-sided significance level (alpha) of 0.05 using a Mann-Whitney test assuming that the actual distributions are equal.RandomizationThe software ��Randlist�� (University of Gottingen, Germany) was used for randomization. Patients were stratified by Acute Physiology and Chronic Health Evaluation III (APACHE III) scores (low risk: ��60; high risk: >60) and random permuted blocks within strata were generated (block size = 6). A person not otherwise involved in this study randomized patients 1:1.IVIG and human albumin were supplied in a form in which no differentiation between verum and placebo was possible. The study medication was linked to the patient numbers for identification according to the randomization list. Participants and care providers were blinded to the treatment.

Statistical methodsData are presented as mean �� standard deviation, median (25th to 75th percentile) or count and relative frequency. Differences between the study groups were assessed using the Fisher��s exact or the Student��s t-test, as appropriate. We performed a number of sensitivity analyses using different metrics for the CIPNM, including the difference from baseline to study end, yielding virtually unchanged results (data not shown). To assess the course of CIPNM we calculated the differences of the CIPNM severity sum scores regardless of the group and compared it versus 0 in a one-sided GSK-3 t-test.