Author: Bc. Marie Hejnová Title: Effect of Functional Electrical Stimulation-Assisted Ergometry on Muscle Cross-Sectional Diameter, Nitrogen and Fluid Balance in Critically Ill Objectives: The aim of this thesis was to investigate the effect of functional electrical stimulation-assisted cycling ergometry (FES-CE) on cross-sectional diameter of the quadriceps femoris (QF) muscle of both lower extremities in critically ill patients. Another objective was to evaluate if the measured values are responding to the changes in muscle tissue or are caused by an oedema. Methods: The intervention group received daily intensified physical therapy and FES-CE. We measured cross-sectional diameter of the QF muscle repeatedly by a diagnostic ultrasound. We recorded daily nitrogen balance to objectivize catabolism of muscle and fluid balance to objectivize amount of cumulative fluid. Results: The total of 115 patients were evaluated. Average decrease of cross-sectional diameter of QF muscle in the intervention group was 0.020 ± 0.070 cm/day, in the control group it was 0.017 ± 0.084 cm/day (p = 0.87). We registered an opposite result from the eighth day onwards, the intervention group had average decrease 0.025 ± 0.047 cm/day and the control group 0.040 ± 0.076 cm/day (p = 0.38). The nitrogen intake was...

Patients admitted to intensive care unit (ICU) are known to develop significant impairments in physical function. Patients with critical illness suffer up to 30% reductions in muscle size within the first ten days of admission to the ICU. Muscle strength testing, Medical Research Council-sum score, is current gold-standard to diagnosis ICU-acquired weakness and predicts risk of mortality and long-term physical function. Muscle power different from muscle strength in that it accounts for velocity of movement, is potentially a better independent predictor of function that has not been studied in this population. In addition, we hypothesize that muscle size and quality measured through ultrasound imaging has better applicability and prediction that strength testing. Therefore, we prospectively collected data surrounding these muscle parameters in patients admitted to the medicine ICU at University of Kentucky. Primary outcomes included physical function, muscle power with a novel assessment tool for the critically ill population, muscle strength, and muscle size and quality assess through ultrasound imaging. 36 patients admitted to ICU and 18 aged-matched controlled were enrolled. Patients had significantly lower scores on muscle power assessment at ICU discharge (33.6 ±19.0 W; t= 4.01, p < 0.001) and at hospital discharge (40.9 ±16.5 W; t= 4.81, p < 0.001) in comparison to controls (59.3± 14.7 W). Patients with better scores on muscle power assessment had significantly better scores on physical function measures (Six-minute walk test; rs = 0.548, p = 0.0001). Muscle size (cross-sectional area of rectus femoris muscle) and muscle power were strongly correlated (rs = 0.66, p < 0.0001). These data suggest that patients with critical illness have significantly reduced muscle power which directly related to deficits in physical function.

Transforming growth factor beta 1 (TGF-beta1) ist ein multifunktionales Zytokin, welches eine Rolle in der Sepsis und in der Sepsis-induzierten Myopathie spielen könnte. Weiterhin könnten erhöhte TGF-beta1-Level zur Muskelschwäche, die mit der Intensivpflege assoziiert ist (engl. intensiv care unit-acquired weakness, ICUAW), beitragen. Der TGF-beta1- Signalweg wurde in Skelettmuskelbiopsien von ICUAW-Patienten heraufreguliert. Secreted frizzled related protein 2 (SFRP2) wurde in einer Gen-Set-Anreicherungsanalyse als das am höchsten regulierte Gen identifiziert. Im Mausmodell führten Sepsis und Hunger zu einer verringerten Sfrp2-Expression, während dies in der Denervation-induzierten Skelettmuskelatrophie nicht festzustellen war. In differenzierten C2C12-Myotuben führte TGF-beta1 zu einer verringerten Sfrp2-mRNA- und Proteinexpression. Luciferase-Assays deuteten auf eine TGF-beta1-abhängige Herunterregulation von Sfrp2 hin, welche auf Promoterebene durch mögliche negative regulatorische Elemente im Sfrp2-Promoter vermittelt wurde. Weiterhin wurde eine TGF-beta1 induzierte Muskelatrophie durch transkriptionelle Repression der myosin heavy chain Gene beobachtet. Im Gegensatz dazu veränderte TGF-beta1 nicht den proteasomalen Abbau muskulärer Proteine. Die Genexpression von Tripartite motif containing 63 und F-box only protein 32 war hingegen leicht herunterreguliert. TGF-beta1-induzierte Atrophie in differenzierten C2C12-Myotuben wurde teilweise durch rekombinantes Sfrp2 aufgehoben. Weiterhin wurde eine direkte physikalische Interaktion zwischen Sfrp2 und TGF-beta1 gefunden, welche diesen Effekt verursacht haben könnte. Zusammengefasst lässt sich feststellen, dass der TGF-beta1- Signalweg eine wichtige Rolle in der ICUAW durch Inhibition der myosin heavy chain Expression spielt. TGF-beta1-abhängige Herunterregulation von Sfrp2 könnte zu einer Feedback-Antwort, die das Ausmaß der Atrophie durch TGF-beta1 verstärkt, führen. / Transforming growth factor beta 1 (TGF-beta1) is a multifunctional cytokine that may play a role in sepsis and in sepsis-induced myopathy. Our group speculated that increased TGF-beta1 could contribute to intensive care (ICU)-acquired weakness (ICUAW), a catastrophic muscle disease in critically ill patients. We found that TGF-beta1 signaling in skeletal muscle biopsies of ICUAW patients was upregulated. Secreted frizzled related protein 2 (SFRP2) was the most regulated gene identified by gene set enrichment analysis (GSEA). I then studied the regulation and function of SFRP2 in different skeletal muscle atrophy models. In three mouse models, downregulated Sfrp2 expression was observed in sepsis and starvation, but not in denervation-induced skeletal muscle atrophy. In differentiated C2C12 myotubes, TGF-beta1 downregulated Sfrp2 expression on both mRNA and protein levels. Luciferase assays suggested that TGF-beta1-dependent downregulation of Sfrp2 was mediated at the promoter level through possible negative regulatory elements in the Sfrp2 promoter. I also observed that TGF-beta1-induced muscle atrophy was accompanied by transcriptional repression of myosin heavy chain genes. In contrast, TGF-beta1 did not increase proteasomal degradation of muscular proteins since gene expression of Tripartite motif containing 63 (Trim63) and F-box only protein (Fbxo32) was not upregulated; instead, they were slightly downregulated. TGF- beta1-induced differentiated C2C12 myotube atrophy was partially reversed by recombinant Sfrp2. This inhibitory effect could have resulted from direct interaction between Sfrp2 and TGF-beta1, since I found a physical interaction between these two proteins. Taken together, TGF-beta1 signaling pathway could play an important role in ICUAW via inhibition of myosin heavy chain expression. TGF-beta1-dependent downregulation of Sfrp2 may establish a feedback loop augmenting the atrophic effect of TGF-beta1.