Mario Fournier

Exercise in Maintenance Hemodialysis Patients Induces Transcriptional Changes in Genes Favoring Anabolic Muscle

Exercise training increases exercise capacity of maintenance hemodialysis patients, but the cellular mechanisms responsible for this effect are unclear. We studied the effects of different forms of exercise training (endurance, strength, or a combination where patients underwent about one-half each

Development of a Diagnostic Method for Detecting Increased Muscle Protein Degradation in Patients with Catabolic Conditions

Muscle atrophy in catabolic illnesses is due largely to accelerated protein degradation. Unfortunately, methods for detecting accelerated muscle proteolysis are cumbersome. The goal of this study was to develop a method for detecting muscle protein breakdown and assess the effectiveness of anticatabolic therapy. In rodent models of catabolic conditions, it was found that accelerated muscle protein degradation is triggered by activation of caspase-3. Caspase-3 cleaves actomyosin/myofibrils to form substrates for the ubiquitin-proteasome system and leaves a characteristic 14-kD actin fragment in the insoluble fraction of a muscle lysate. Muscle biopsies were obtained from normal adults and three groups of patients: 14 who were undergoing hip arthroplasty, 28 hemodialysis patients who were participating in exercise programs, and seven severely burned patients. In muscle of patients who were undergoing hip arthroplasty, the 14-kD actin fragment level was correlated (r = 0.787, P < 0.01) with the fractional rate of protein degradation. In muscle of hemodialysis patients who were undergoing endurance exercise training, the 14-kD actin fragment decreased to values similar to levels in normal adults; strength training did not significantly decrease the actin fragment. Severely burned patients had increased muscle protein degradation and actin fragment levels, but the two measures were not significantly correlated. The experimental results suggest that the 14-kD actin fragment in muscle biopsies is increased in catabolic states and could be used in conjunction with other methods to detect and monitor changes in muscle proteolysis that occur in patients with mild or sustained increases in muscle proteolysis.

Neuromuscular transmission failure during postnatal development

Neuromuscular transmission failure in rat diaphragm muscle was examined during the first two weeks of postnatal development, and was found to contribute significantly more to diaphragm fatigue induced by repetitive activation than in adult muscle. Intracellular analysis of evoked end-plate potentials indicated that neuromuscular transmission failure was due to both a block of action potential propagation and reduced synaptic efficacy.

Time course expression of Foxo transcription factors in skeletal muscle following corticosteroid administration

Increased expression of forkhead box O (Foxo) transcription factors were reported in cultured myotubes and mouse limb muscle with corticosteroid (CS) treatment. We previously reported that administration of CS to rats resulted in muscle fiber atrophy only by day 7. The aim of this study, therefore, was to evaluate the time-course changes in the expression of Foxo transcription factors and muscle-specific ubiquitin E3 ligases in rat limb muscle following CS administration. Triamcinolone (TRI; 1 mg x kg(-1) x day(-1) im) was administered for 1, 3, or 7 days. Control (CTL) rats were given saline. Muscle mRNA was analyzed by real-time RT-PCR. Compared with CTL, body weights of TRI-treated animals decreased by 3, 12, and 21% at days 1, 3, and 7, respectively. Muscle IGF-1 mRNA levels decreased by 33, 65, and 58% at days 1, 3, and 7 in TRI-treated rats compared with CTL. Levels of phosphorylated Akt were 28, 50, and 36% lower in TRI animals at these time points. Foxo1 mRNA increased progressively by 1.2-, 1.4-, and 2.5-fold at days 1, 3, and 7 in TRI animals. Similar changes were noted in the expression of Foxo3a mRNA (1.3-, 1.4-, and 2.6-fold increments). By contrast, Foxo4 mRNA was not significantly changed in TRI animals. With TRI, muscle atrophy F box/Atrogin-1 increased by 1.8-, 4.1-, and 7.5-fold at days 1, 3, and 7 compared with CTL rats. By contrast, muscle RING finger 1 increased only from day 7 (2.7-fold). Gradual reduction in IGF-I expression with TRI over the time series paralleled that of Akt. These findings are consistent with a progressive stimulus to muscle protein degradation and the need to process/remove disassembled muscle proteins via the ubiquitin-proteasome system. Elucidating the dynamic catabolic responses to CS challenge is important in understanding the mechanisms underlying muscle atrophy and therapeutic measures to offset this.

Testosterone and resistance training effects on muscle nitric oxide synthase isoforms in COPD men

BACKGROUND Skeletal muscle dysfunction contributes to exercise limitation in COPD. The role of the nitric oxide synthase (NOS) system in muscle dysfunction is ill defined. Reduced levels of endothelial NOS (eNOS) and elevated levels of inducible NOS (iNOS) in the skeletal muscle of COPD patients have been recently reported. We hypothesized that resistance exercise training (R) and/or testosterone supplementation (T) would alter the transcription and expression of the NOS isoenzymes in COPD skeletal muscle. METHODS Vastus lateralis biopsies were obtained before and after a 10-week intervention in 40 men with severe COPD(age 67.7 ± 8.3, FEV(1) 41.4 ± 12.6% predicted): placebo + no training (P) (n = 11), placebo + resistance training (PR) (n = 8), testosterone + no training (T) (n = 11) and testosterone + resistance training (TR) (n = 10) groups. eNOS, nNOS and iNOS mRNA and protein levels were measured in each sample. mRNA and protein levels were measured using real-time PCR and enzyme-linked immunosorbant assay, respectively. RESULTS eNOS mRNA increased in the TR group compared to P and T groups (P < 0.001). eNOS protein was increased in TR and T groups after intervention (P < 0.05) but not in the PR group. nNOS protein increased in the PR, T, and TR groups (P < 0.05). iNOS protein decreased only in the TR group (P = 0.01). CONCLUSION Resistance training and testosterone supplementation increased eNOS and nNOS proteins and decreased iNOS protein in the skeletal muscles of men with COPD. These changes in NO system might explain some of the favorable effects of these therapies.

Metabolic and morphometric profile of muscle fibers in chronic hemodialysis patients

Muscle weakness and effort intolerance are common in maintenance hemodialysis (MHD) patients. This study characterized morphometric, histochemical, and biochemical properties of limb muscle in MHD patients compared with controls (CTL) with similar age, gender, and ethnicity. Vastus lateralis muscle biopsies were obtained from 60 MHD patients, 1 day after dialysis, and from 21 CTL. Muscle fiber types and capillaries were identified immunohistochemically. Individual muscle fiber cross-sectional areas (CSA) were quantified. Individual fiber oxidative capacities were determined (microdensitometric assay) to measure succinate dehydrogenase (SDH) activity. Mean CSAs of type I, IIA, and IIX fibers were 33, 26, and 28% larger in MHD patients compared with CTL. SDH activities for type I, IIA, and IIX fibers were reduced by 29, 40, and 47%, respectively, in MHD. Capillary to fiber ratio was increased by 11% in MHD. The number of capillaries surrounding individual fiber types were also increased (type I: 9%; IIA: 10%; IIX: 23%) in MHD patients. However, capillary density (capillaries per unit muscle fiber area) was reduced by 34% in MHD patients, compared with CTL. Ultrastuctural analysis revealed swollen mitochondria with dense matrix in MHD patients. These results highlight impaired oxidative capacity and capillarity in MHD patients. This would be expected to impair energy production as well as substrate and oxygen delivery and exchange and contribute to exercise intolerance. The enlarged CSA of muscle fibers may, in part, be accounted for by edema. We speculate that these changes contribute to reduce limb strength in MHD patients by reducing specific force.

Arterial injury in mice with severe insulin-like growth factor-1 (IGF-1) deficiency.

Background: Insulin-like growth factor-1 plays a significant role in wound healing. Injury to the arterial wall is followed by a marked increase in insulin-like growth factor-1 expression and inhibition of insulin-like growth factor-1 action is associated with diminished intimal thickening after injury. Methods and Results: The role of insulin-like growth factor-1 in arterial response to cuffinjury was investigated in genetically modified mice with severe insulin-like growth factor-1 deficiency (m/m mice). Tissue and serum insulin-like growth factor-1 was severely decreased, by 40% to 60% before the injury and by 50% to 60% following the arterial injury in insulin-like growth factor-1 m/m mice compared to control mice. Nevertheless, following the cuff induced injury to the carotid arteries, insulin-like growth factor-1m/m mice had a similar number of proliferating medial cells 3 days after injury and similar neointimal thickening (0.019 ± 0.015 C57BL/6J vs. 0.016 ± 0.014 mm2, P = 0.26) 21 days after injury compared to wild type C57BL/6J mice. The phases of the response to injury that are mediated by insulin-like growth factor-1 were studied with recombinant human insulin-like growth factor-1 in rats with balloon-injured femoral arteries. Treatment of rats with recombinant human insulin-like growth factor-1 increased neointimal thickening (0.0265 ± 0.0099 vs 0.0156 ± 0.0049 mm2, P = 0.03), intimal smooth muscle cell numbers (195.6 ± 40.2 vs 145.3±27.3; P = 0.03), and the ratio of proliferating intimal to medial smooth muscle cells (10.7 ± 6.9 vs 3.0 ± 2.1; P = 0.03) 7 days after injury compared to untreated rats. At 14 days neointimal area was similar in the 2 groups of rats. Conclusions: The data in insulin-like growth factor-1 deficient mice suggest a relatively low threshold tissue concentration for insulin-like growth factor-1 to exhibit its role in vascular response to injury. The findings in rats treated with recombinant human insulin-like growth factor-1 suggest that insulin-like growth factor-1 is primarily involved in the early phases of neointimal formation.

Respiratory work in elastase treated hamsters

Biomechanical adaptations of the diaphragm in the hamster model of emphysema are similar to those observed in skeletal muscle with exercise training. The aim of this study was to evaluate whether the dynamic pressure-volume (PV) work of breathing in hamsters with elastase-induced emphysema may contribute to these adaptations. PV work in elastase treated animals was compared to healthy controls. The studies were performed in adult hamsters 14-16 months following intratracheal administration of elastase (elastase treated group, ET) or saline (control group, CTL). Airway and esophageal pressures and air flows were measured during spontaneous breathing in anesthetized, supine animals. Pulmonary work (WL) was computed from transpulmonary pressures and airflows. Functional residual capacity (FRC) and total lung capacity (TLC; defined as volume at 25 cmH2O) in ET were increased 2 and 1.8 times, respectively, compared with CTL. Averaged tidal volume (VT) and inspiratory flows were comparable between groups. Total work of breathing (WT) normalized per ml VT was not significantly affected with elastase treatment but the pulmonary elastance work (WE) was significantly less in ET animals than controls (0.88 +/- 0.61 g cm(-2) vs. 1.63 +/- 0.32). Pulmonary resistive work was not significantly different between ET and CTL animals. These results suggest that biomechanical adaptations of the diaphragm observed in ET hamsters are caused by mechanisms other than the changes in dynamic mechanical properties of the lung following elastase treatment.

Exosome-Mediated Benefits of Cell Therapy in Mouse and Human Models of Duchenne Muscular Dystrophy

Summary Genetic deficiency of dystrophin leads to disability and premature death in Duchenne muscular dystrophy (DMD), affecting the heart as well as skeletal muscle. Here, we report that clinical-stage cardiac progenitor cells, known as cardiosphere-derived cells (CDCs), improve cardiac and skeletal myopathy in the mdx mouse model of DMD. Injection of CDCs into the hearts of mdx mice augments cardiac function, ambulatory capacity, and survival. Exosomes secreted by human CDCs reproduce the benefits of CDCs in mdx mice and in human induced pluripotent stem cell-derived Duchenne cardiomyocytes. Surprisingly, CDCs and their exosomes also transiently restored partial expression of full-length dystrophin in mdx mice. The findings further motivate the testing of CDCs in Duchenne patients, while identifying exosomes as next-generation therapeutic candidates.

Exercise capacity in hamsters with elastase-induced emphysema compared to normal controls.

The purpose of this study was to determine whether hamsters with elastase-induced emphysema (EMP) would demonstrate a reduction in exercise capacity compared to control (CON) hamsters and whether changes in activity levels, muscle function and structure could explain any changes in exercise capacity. Peak oxygen consumption and daily activity levels were measured on two occasions. Inspiratory capacity under deep anesthesia, in vitro measurements of muscle force and fatigability for the diaphragm (DIA) and extensor digitorum longus (EDL) and fiber proportions, muscle cross-sectional area and fiber specific SDH activity from the DIA, EDL and vastus lateralis (VLA) were obtained. Inspiratory capacity was 60% higher in the EMP compared to CON hamsters (p=0.0004). Activity levels and exercise capacity were not significantly different between EMP and CON hamsters. Muscle strength and fatigability, fiber proportions, muscle cross-sectional area and fiber specific SDH activity were similar between EMP and CON hamsters. In conclusion, in hamsters, elastase-induced emphysema did not reduce maximal exercise capacity.