Erythropoietin Concentration in Boys With p.His63Asp Polymorphism of the HFE Gene.

The molecular mechanism that regulates iron homeostasis is based on a network of signals, which reflect on the iron requirements of the body. HFE-related hemochromatosis is characterized by excessive intestinal absorption of dietary iron, in particular cases resulting in pathologically high iron storage in tissues and organs. During childhood, HFE gene homozygosity or heterozygosity manifests exclusively in the form of biochemical abnormalities. Because of their mutual link, bioavailable iron and endogenous erythropoietin (EPO) are indispensable for effective erythropoiesis. We analyzed the impact of p.(His63Asp) polymorphism of the HFE gene on erythropoiesis taking into consideration endogenous EPO production in the developmental age. In the study we performed, we observed a significant, strong and negative correlation between the concentration of EPO, hemoglobin, and red blood cell count. A negative trend was also noted on the impact of iron concentration and transferrin saturation on EPO production. In conclusion, this preliminary study demonstrates an impaired impact of endogenous EPO on erythropoiesis in the presence of increased iron content in carriers of p.(His63Asp) (heterozygotes) variant of the HFE gene in developmental age.

HFE Gene Mutations and Iron Status in 100 Healthy Polish Children.

Iron participates in oxygen transport, energetic, metabolic, and immunologic processes. There are 2 main causes of iron overload: hereditary hemochromatosis which is a primary cause, is a metabolic disorder caused by mutations of genes that control iron metabolism and secondary hemochromatosis caused by multitransfusions, chronic hemolysis, and intake of iron rich food. The most common type of hereditary hemochromatosis is caused by HFE gene mutation. In this study, we analyzed iron metabolism in 100 healthy Polish children in relation to their HFE gene status. The wild-type HFE gene was predominant being observed in 60 children (60%). Twenty-five children (25%), presented with heterozygotic H63D mutation, and 15 children (15%), presented with other mutations (heterozygotic C282Y and S65C mutation, compound heterozygotes C282Y/S65C, C282Y/H63D, H63D homozygote). The mean concentration of iron, the level of ferritin, and transferrin saturation were statistically higher in the group of HFE variants compared with the wild-type group. H63D carriers presented with higher mean concentration of iron, ferritin levels, and transferrin saturation compared with the wild-type group. Male HFE carriers presented with higher iron concentration, transferrin saturation, and ferritin levels than females. This preliminary investigation demonstrates allelic impact on potential disease progression from childhood.

Repeated "all out" interval exercise causes an increase in serum hepcidin concentration in both trained and untrained men.

This investigation assessed effects of high-intensity interval exercise (HIE; triple Wingate anaerobic test) on inflammatory markers, iron metabolism and hepcidin concentrations. Group of highly trained judo athletes (TR) and non-trained control males (CG) completed a triple Wingate test separated by 4.5min rest. Venous blood samples were collected before, immediately after, 1h, 24h, and 5days following exercise and analysed for serum of IL-6, IL-10, iron, and ferritin. Physiological response to exercise was also determined. Concentration of IL-6 and hepcidin increased 1h after exercise in both groups (p<0.05). Hepcidin returned post testing 24h in TR, whereas in CG it remained elevated during 5days following exercise. Changes in hepcidin did not correlate with shifts in serum IL-6, iron and ferritin concentrations. Gathered data suggest that following HIE, hepcidin increased independently of IL-6 and neither blood nor storage iron affected this phenomena.

The Rehabilitation Program Improves Balance Control in Children with Excessive Body Weight and Flat Feet by Activating the Intrinsic Muscles of the Foot: A Preliminary Study.

BACKGROUND: determining the appropriate rehabilitation protocol is essential to influence the correction of flat feet, e.g., by activating the intrinsic muscles of the foot. Therefore, this study aimed to determine the impact of the exercises activating the intrinsic foot muscles for postural control in children with flat feet, with normal and excessive body weight. METHODS: Fifty-four children aged 7 to 12 were enrolled in the research. Forty-five children were qualified for the final evaluation. Each child in the experimental group was demonstrated an appropriate technique for performing a short foot exercise without compensation by extrinsic muscle. The participants then performed a supervised short foot training session once a week and on other days of the week under the supervision of caregivers for 6 weeks. Flat feet were scored on the foot posture index scale. A postural test was evaluated with a Biodex balance system SD. Statistical significance in the foot posture index scale and postural test were evaluated using an analysis of variance (ANOVA) with Tukey's post-hoc test. RESULTS: according to the six indices of the foot posture index scale, five indicators showed statistically significant improvement after rehabilitation. At the 8-12 platform mobility level, it was revealed that the excessive body weight group had significant improvements in the overall stability index and medio-lateral stability index, with eyes closed. CONCLUSION: our results indicate that a 6-week rehabilitation program based on the activation of the intrinsic muscles of the foot resulted in an improvement in the foot position. This, in turn, affected balance control, especially in children with excess body weight in conditions of closed eyes.

Exercise training enhances the skeletal muscle response to radiation-induced oxidative stress.

Overproduction of reactive oxygen species (ROS) can damage cellular macromolecules and lead to cellular dysfunction or death. Exercise training induces beneficial adaptations in skeletal muscle that may reduce cellular damage from exposure to ROS. To determine the response of exercise-conditioned muscle to acute increases in ROS, four groups of mice were used: non-trained (NT, n = 12); NT + high-dose radiation (HDR, n = 3); exercise-trained (EX, n = 13, 3 days/week for 10 weeks, 10 m/min to 18 m/min); and EX + HDR (n = 3/group). Quadriceps muscle was harvested 3-5 days following the last exercise bout in the training program for measurement of antioxidant enzyme and metabolic enzyme activity. Total superoxide dismutase (41%), and manganese sodium oxide dismutase (51%) activities were significantly increased in radiation-challenged EX mice as compared with unchallenged EX mice (all P ≤ 0.05). No such increase was observed in NT mice. Citrate synthase (42%) and cytochrome c oxidase (38%) activities were both elevated in radiation-challenged EX mice as compared with unchallenged EX mice (both P < 0.05), and no such increase was observed in NT. We demonstrate that preconditioning skeletal muscle with EX enhances the response of antioxidant and mitochondrial enzymes to radiation.

Early Rehabilitation Program and Vitamin D Supplementation Improves Sensitivity of Balance and the Postural Control in Patients after Posterior Lumbar Interbody Fusion: A Randomized Trial.

BACKGROUND: The introduction of early rehabilitation exercise is the foundation of treatment post-Posterior lumbar interbody fusion (PLIF) surgery, and the search for additional sources of reinforcement physiotherapy seems to be very important. METHODS: The patients were randomly divided into the vitamin D3 (n = 15; D3) supplemented group and received 3200 IU per day for five weeks before surgery and the placebo group (n = 18; Pl) received vegetable oil during the same time. The patients began the supervisor rehabilitation program four weeks after surgery. RESULTS: The limits of stability (LOS) were significantly improved in the D3 group after 5 and 14 weeks (p < 0.05), while in the Pl group, progress was only observed after 14 weeks (p < 0.05). The LOS were also higher in the D3 group than in the Pl group after five weeks of supervised rehabilitation (p < 0.05). In the postural stability (PST) test, significant progress was observed in the D3 group after 14 weeks (p < 0.02). In addition, neither rehabilitation nor supplementation had significant effects on the risk of falls (RFT). CONCLUSIONS: Vitamin D supplementation seems to ameliorate the effects of an early postoperative rehabilitation program implemented four weeks after posterior lumbar interbody fusion. Early physiotherapy treatment after PLIF surgery combined with vitamin D supplementation appears to be a very important combination with regard to the patients' recovery process.

The effects of gymnastics training on selected parameters of anaerobic capacity in 12-year-old boys.

BACKGROUND: One of the aims of the study was to describe the physiological factors of young boys participating in artistic gymnastic training and evaluate differences between the levels of aerobic and anaerobic efficiency in this group as compared to the control group. METHODS: The young male gymnasts selected to participate in the present study have been (G) participating in the training process since they were 6 years old (N.=12, age 11-12 y). In the control group (N.=12, age 11-12 y), boys were participating in physical education classes (C). Anaerobic efficiency was evaluated by using a 30 s Wingate Anaerobic Test (WAnT) for arms. Adjusted load was defined on the level of 50 g·kg-1 of body mass. Aerobic efficiency was defined using gradual effort to exhaustion for lower limbs on the cycle ergometer with simultaneous analysis of breathing gases. RESULTS: The test to exhaustion showed that group G achieved lower VO2peak results as compared to group C. The values were respectively: 48.1 mL·kg-1·min-1 and 55.6 mL·kg-1·min-1 (P<0.05). In the examination of anaerobic efficiency for upper limb parameters, total work and mean power were higher in group G than in group C, while the fatigue index (FI) was lower. CONCLUSIONS: It is likely that early specialization in young male gymnasts may influence proper aerobic metabolism development. Executing WAnT using arms in group G was more convenient and precise according to anaerobic efficiency.

hmSOD1 gene mutation-induced disturbance in iron metabolism is mediated by impairment of Akt signalling pathway.

BACKGROUND: Recently, skeletal muscle atrophy, impairment of iron metabolism, and insulin signalling have been reported in rats suffering from amyotrophic lateral sclerosis (ALS). However, the interrelationship between these changes has not been studied. We hypothesize that an impaired Akt-FOXO3a signalling pathway triggers changes in the iron metabolism in the muscles of transgenic animals. METHODS: In the present study, we used transgenic rats bearing the G93A hmSOD1 gene and their non-transgenic littermates. The study was performed on the muscles taken from animals at three different stages of the disease: asymptomatic (ALS I), the onset of the disease (ALS II), and the terminal stage of the disease (ALS III). In order to study the molecular mechanism of changes in iron metabolism, we used SH-SY5Y and C2C12 cell lines stably transfected with pcDNA3.1, SOD1 WT and SOD1 G93A, or FOXO3a TM-ER. RESULTS: A significant decrease in P-Akt level and changes in iron metabolism were observed even in the group of ALS I animals. This was accompanied by an increase in the active form of FOXO3a, up-regulation of atrogin-1, and catalase. However, significant muscle atrophy was observed in ALS II animals. An increase in ferritin L and H was accompanied by a rise in PCBP1 and APP protein levels. In SH-SY5Y cells stably expressing SOD1 or SOD1 G93A, we observed elevated levels of ferritin L and H and non-haem iron. Interestingly, insulin treatment significantly down-regulated ferritin L and H proteins in the cell. Conversely, cells transfected with small interfering RNA against Akt 1, 2, 3, respectively, showed a significant increase in the ferritin and FOXO3a levels. In order to assess the role of FOXO3a in the ferritin expression, we constructed a line of SH-SY5Y cells that expressed a fusion protein made of FOXO3a fused at the C-terminus with the ligand-binding domain of the oestrogen receptor (TM-ER) being activated by 4-hydroxytamoxifen. Treatment of the cells with 4-hydroxytamoxifen significantly up-regulated ferritin L and H proteins level. CONCLUSIONS: Our data suggest that impairment of insulin signalling and iron metabolism in the skeletal muscle precedes muscle atrophy and is mediated by changes in Akt/FOXO3a signalling pathways.

Low intensity training decreases markers of oxidative stress in skeletal muscle of mdx mice.

Reactive oxygen species may contribute to the pathogenesis of muscular dystrophy. High intensity exercise clearly induces muscle damage in mdx mice; however, the effects of low intensity exercise training (LIT) on mdx muscle are less clear. We examined the effect of LIT on markers of oxidative stress (malondialdehyde and protein carbonyls), antioxidant (superoxide dismutase, catalase, and glutathione peroxidase), and mitochondrial (2-oxoglutarate dehydrogenase and cytochrome oxidase) enzymes in skeletal muscle of mdx and wild-type mice. Mdx and wild-type mice were allocated to LIT and sedentary groups. Malondialdehyde levels were higher in white muscle from sedentary mdx as compared to both sedentary and LIT wild-type mice (P<0.001). Protein carbonyl content was higher in white and red muscle of mdx versus wild-type mice (P<0.05). LIT was associated with lower levels of malondialdehyde and protein carbonyls in white muscle of mdx mice (decreased 38 and 44%, P<0.001 and P<0.01, respectively). Antioxidant and mitochondrial enzyme activities were higher in white muscle of mdx than in wild-type mice (P<0.05). LIT in mdx mice induced physiological adaptation resulting in lower levels of markers of oxidative stress that were not different than those from wild type. These results are of relevance for therapeutic exercise in patients with dystrophinopathy where exercise prescription remains controversial.

Higher oxidative stress in skeletal muscle of McArdle disease patients.

McArdle disease (MCD) is an autosomal recessive condition resulting from skeletal muscle glycogen phosphorylase deficiency. The resultant block in glycogenolysis leads to an increased flux through the xanthine oxidase pathway (myogenic hyperuricemia) and could lead to an increase in oxidative stress. We examined markers of oxidative stress (8-isoprostane and protein carbonyls), NAD(P)H-oxidase, xanthine oxidase and antioxidant enzyme (superoxide dismutase, catalase and glutathione peroxidase) activity in skeletal muscle of MCD patients (N = 12) and controls (N = 12). Eight-isoprostanes and protein carbonyls were higher in MCD patients as compared to controls (p < 0.05). There was a compensatory up-regulation of catalase protein content and activity (p < 0.05), mitochondrial superoxide dismutase (MnSOD) protein content (p < 0.01) and activity (p < 0.05) in MCD patients, yet this increase was not sufficient to protect the muscle against elevated oxidative damage. These results suggest that oxidative stress in McArdle patients occurs and future studies should evaluate a potential role for oxidative stress contributing to acute pathology (rhabdomyolysis) and possibly later onset fixed myopathy.

Long-term caloric restriction abrogates the age-related decline in skeletal muscle aerobic function.

The purpose of this study was to determine the effect of long-term caloric restriction (CR) on the age-associated decline of skeletal muscle aerobic function. Skeletal muscle maximal aerobic performance (VO2max) was assessed in ad libitum (AL) and CR rats aged 8-10 months and 35 months using a pump-perfused hindlimb model to match oxygen delivery to muscle mass between groups. Whereas there was a 46% decline in muscle mass-specific VO2max between 8-10 mo (524+/-13 micromol x min(-1) x 100 g(-1); mean+/- SE) and 35 mo (281+/-54 micromol x min(-1) x 100 g(-1)) in AL rats, not only did CR rats begin at the same point in 8-10 mo old rats (490+/-42 micromol x min(-1) x 100 g(-1)), we found no decline in 35 mo old CR animals (484+/-49 micromol x min(-1) x 100 g(-1)). Interestingly, although most markers of oxidative capacity began at a lower point in young adult CR animals, CR rats exhibited a higher in situ activity of complex IV at VO2max. This activity allows the young adult CR animals to exhibit normal aerobic capacity despite the lower oxidative enzyme activities. In stark contrast to the 19-41% decline in activities of citrate synthase, complexes I-III, and complex IV in homogenates prepared from the plantaris muscle and mixed region of gastrocnemius muscle with aging in AL rats, no age-related decline was found in CR animals. Thus, our results showed that CR preserves aerobic function in aged skeletal muscles by facilitating a higher in situ function of complex IV and by preventing the age-related decline in mitochondrial oxidative capacity.

The effect of aging on anaerobic and aerobic enzyme activities in human skeletal muscle.

The effect of aging on metabolic enzyme activity remains controversial, possibly due to physical activity differences. We examined the effect of aging on the enzyme activity for anaerobic and aerobic pathways in nonweight-bearing human skeletal muscle from relatively sedentary males. The muscle obliquus internus abdominis was analyzed for anaerobic (creatine kinase, adenylate kinase, and lactate dehydrogenase) and aerobic (2-oxoglutarate dehydrogenase and carnitine palmitoyltransferase) enzyme activities in two groups: middle-aged (29-54 years) and older (61-74 years) adults. All enzyme activities were lower in older versus middle-aged adults when results were expressed as muscle wet weight (p <.05). When activity was expressed relative to the protein content, only lactate dehydrogenase remained significantly lower in older versus middle-aged adults (p <.001). In conclusion, some of the reduction in muscle performance in older adults may be due to lower activity of the anaerobic and aerobic enzymes as well as protein content, not solely due to a decrease in physical activity.

Antioxidant enzyme activity is up-regulated after unilateral resistance exercise training in older adults.

Cellular antioxidant capacity and oxidative stress are postulated to be critical factors in the aging process. The effects of resistance exercise training on the level of skeletal muscle oxidative stress and antioxidant capacity have not previously been examined in older adults. Muscle biopsies from both legs were obtained from the vastus lateralis muscle of 12 men 71 +/- 7 years of age. Subjects then engaged in a progressive resistance exercise-training program with only one leg for 12 weeks. After 12 weeks, the nontraining leg underwent an acute bout of exercise (exercise session identical to that of the trained leg at the same relative intensity) at the same time as the last bout of exercise in the training leg. Muscle biopsies were collected from the vastus lateralis of both legs 48 h after the final exercise bout. Electron transport chain enzyme activity was unaffected by resistance training and acute resistance exercise (p < 0.05). Training resulted in a significant increase in CuZnSOD (pre--7.2 +/- 4.2, post--12.6 +/- 5.6 U.mg protein(-1); p = 0.02) and catalase (pre--8.2 +/- 2.3, post--14.9 +/- 7.6 micromol.min(-1).mg protein(-1); p = 0.02) but not MnSOD activity, whereas acute exercise had no effect on the aforementioned antioxidant enzyme activities. Furthermore, basal muscle total protein carbonyl content did not change as a result of exercise training or acute exercise. In conclusion, unilateral resistance exercise training is effective in enhancing the skeletal muscle cellular antioxidant capacity in older adults. The potential long-term benefits of these adaptations remain to be evaluated.

Neutrophil engagement and septic challenge in acute experimental pancreatitis in rats.

AIM: To investigate the influence of neutrophil adhesion molecule blockade with monoclonal antibody (MoAb CD11b) and E. coli lipopolysaccharide (LPS) administration on experimental acute pancreatitis (AP). METHODS: AP was induced by four ip injections of cerulein (Cn) at 1-h intervals. MoAb CD 11b and LPS were administered at the beginning of the experiment. RESULTS: The neutrophil count and chemiluminescence were diminished at the beginning of AP. The oxidative stress parameters were found within the pancreatic gland. MoAb CD 11b used for AP resulted in a significant reduction of pancreatic infiltration and pancreatitis oxidative stress parameters. Serum interleukin-6 (IL-6) was not detected in AP animals, whereas high serum IL-6 concentration was noted only in animals receiving LPS. CONCLUSION: Neutrophils are involved in pancreatic damage in the early stage of AP. Neutrophil infiltration reduction protects the pancreatic gland from destruction during AP. LPS does not change the early course of Cn pancreatitis in rats.

Whole-body cryostimulation as an effective way of reducing exercise-induced inflammation and blood cholesterol in young men.

Inflammation may accompany obesity and a variety of diseases, or result from excessive exercise. The aim of this study was to investigate the anti-inflammatory effect of whole-body cryostimulation on the inflammatory response induced by eccentric exercise under laboratory conditions. The study also sought to establish if cold treatment changes the lipid profile and modifies energy expenditure in young people. Eighteen healthy and physically active, college-aged men volunteered to participate in the experiment. They were divided into two subgroups: CRY- submitted to whole-body cryostimulation, and CONT- a control group. Both groups performed eccentric work to induce muscle damage. Blood samples were collected before and 24 h after the exercise. Over the five days that followed, the CRY group was exposed to a series of 10 sessions in a cryogenic chamber (twice a day, for 3 min, at a temperature of -110̊C). After this period of rest, both groups repeated a similar eccentric work session, following the same schedule of blood collection. The perceived pain was noted 24h after each session of eccentric workout. A 30-minute step up/down work-out induced delayed-onset muscle soreness in both groups. The five-day recovery period accompanied by exposure to cold significantly enhanced the concentration of the anti-inflammatory cytokine IL-10. It also led to a pronounced reduction in levels of the pro-inflammatory cytokine IL-1ß, and reduced muscle damage. The values for IL-10 before the second bout of eccentric exercise in the CRY group were 2.0-fold higher in comparison to baseline, whereas in the CONT group, the concentration remained unchanged. Furthermore, blood concentrations of the pro-inflammatory cytokine IL-1ß fell significantly in the CRY group. The main finding of this study was that a series of 10 sessions of whole body cryostimulation significantly reduced the inflammatory response induced by eccentric exercise. The lipid profile was also improved, but there was no effect on energy expenditure during the exercise.

Effect of ethyl pyruvate on skeletal muscle metabolism in rats fed on a high fat diet.

Impaired mitochondrial capacity may be implicated in the pathology of chronic metabolic diseases. To elucidate the effect of ethyl pyruvate supplementation on skeletal muscles metabolism we examined changes in activities of mitochondrial and antioxidant enzymes, as well as sulfhydryl groups oxidation (an indirect marker of oxidative stress) during the development of obesity. After 6 weeks feeding of control or high fat diet, Wistar rats were divided into four groups: control diet, control diet and ethyl pyruvate, high fat diet, and high fat diet and ethyl pyruvate. Ethyl pyruvate was administered as 0.3% solution in drinking water, for the following 6 weeks. High fat diet feeding induced the increase of activities 3-hydroxyacylCoA dehydrogenase, citrate synthase, and fumarase. Moreover, higher catalase and superoxide dismutase activities, as well as sulfhydryl groups oxidation, were noted. Ethyl pyruvate supplementation did not affect the mitochondrial enzymes' activities, but induced superoxide dismutase activity and sulfhydryl groups oxidation. All of the changes were observed in soleus muscle, but not in extensor digitorum longus muscle. Additionally, positive correlations between fasting blood insulin concentration and activities of catalase (p = 0.04), and superoxide dismutase (p = 0.01) in soleus muscle were noticed. Prolonged ethyl pyruvate consumption elevated insulin concentration, which may cause modifications in oxidative type skeletal muscles.

Aberrant mitochondrial homeostasis in the skeletal muscle of sedentary older adults.

The role of mitochondrial dysfunction and oxidative stress has been extensively characterized in the aetiology of sarcopenia (aging-associated loss of muscle mass) and muscle wasting as a result of muscle disuse. What remains less clear is whether the decline in skeletal muscle mitochondrial oxidative capacity is purely a function of the aging process or if the sedentary lifestyle of older adult subjects has confounded previous reports. The objective of the present study was to investigate if a recreationally active lifestyle in older adults can conserve skeletal muscle strength and functionality, chronic systemic inflammation, mitochondrial biogenesis and oxidative capacity, and cellular antioxidant capacity. To that end, muscle biopsies were taken from the vastus lateralis of young and age-matched recreationally active older and sedentary older men and women (N = 10/group; female symbol = male symbol). We show that a physically active lifestyle is associated with the partial compensatory preservation of mitochondrial biogenesis, and cellular oxidative and antioxidant capacity in skeletal muscle of older adults. Conversely a sedentary lifestyle, associated with osteoarthritis-mediated physical inactivity, is associated with reduced mitochondrial function, dysregulation of cellular redox status and chronic systemic inflammation that renders the skeletal muscle intracellular environment prone to reactive oxygen species-mediated toxicity. We propose that an active lifestyle is an important determinant of quality of life and molecular progression of aging in skeletal muscle of the elderly, and is a viable therapy for attenuating and/or reversing skeletal muscle strength declines and mitochondrial abnormalities associated with aging.

Limb immobilization induces a coordinate down-regulation of mitochondrial and other metabolic pathways in men and women.

Advancements in animal models and cell culture techniques have been invaluable in the elucidation of the molecular mechanisms that regulate muscle atrophy. However, few studies have examined muscle atrophy in humans using modern experimental techniques. The purpose of this study was to examine changes in global gene transcription during immobilization-induced muscle atrophy in humans and then explore the effects of the most prominent transcriptional alterations on protein expression and function. Healthy men and women (N = 24) were subjected to two weeks of unilateral limb immobilization, with muscle biopsies obtained before, after 48 hours (48 H) and 14 days (14 D) of immobilization. Muscle cross sectional area (approximately 5%) and strength (10-20%) were significantly reduced in men and women (approximately 5% and 10-20%, respectively) after 14 D of immobilization. Micro-array analyses of total RNA extracted from biopsy samples at 48 H and 14 D uncovered 575 and 3,128 probes, respectively, which were significantly altered during immobilization. As a group, genes involved in mitochondrial bioenergetics and carbohydrate metabolism were predominant features at both 48 H and 14 D, with genes involved in protein synthesis and degradation significantly down-regulated and up-regulated, respectively, at 14 D of muscle atrophy. There was also a significant decrease in the protein content of mitochondrial cytochrome c oxidase, and the enzyme activity of cytochrome c oxidase and citrate synthase after 14 D of immobilization. Furthermore, protein ubiquitination was significantly increased at 48 H but not 14 D of immobilization. These results suggest that transcriptional and post-transcriptional suppression of mitochondrial processes is sustained throughout 14 D of immobilization, while protein ubiquitination plays an early but transient role in muscle atrophy following short-term immobilization in humans.

Increased PFK activity and GLUT4 protein content in McArdle's disease.

Inborn errors of metabolism represent an opportunity to conduct studies in order to understand compensatory adaptations to a defective metabolic pathway. We evaluated the molecular and biochemical adaptations in substrate metabolism (glycolysis, electron transport chain, tricarboxylic acid cycle, beta-oxidation) in response to myophosphorylase deficiency in skeletal muscle from 13 patients with McArdle's disease (MD) and 13 age-matched controls. MD muscle had higher phosphofructokinase protein content and activity as well as glucose transporter 4 (GLUT4) protein content and lower GLUT4 mRNA content than controls. At the protein level, skeletal muscle adaptations suggest an augmented glucose transport and glycolytic flux as a compensatory metabolic strategy to a chronic absence of muscle glycogen phosphorylase. These results support previous findings of increased glucose uptake during exercise and alleviation of symptoms with oral sucrose in patients with MD.

Effects of a CRF2R agonist and exercise on mdx and wildtype skeletal muscle.

Corticotrophin-releasing factor 2 receptor (CRF2R) agonists prevent muscle atrophy due to immobilization, denervation, and corticosteroid-induced muscle atrophy in wildtype mice. We hypothesized that a CRF2R agonist will increase skeletal muscle mass in mdx mice. Mdx (C57BL/10ScSn-Dmd(mdx)) and wildtype (C57BL/6) mice were divided into four groups: sedentary placebo, sedentary CRF2R agonist, exercised placebo, and exercised CRF2R agonist. Mice exercised on a treadmill twice weekly for 30 min (8-12 m/min, 8 weeks). Muscle and heart weights, serum creatine kinase, and gamma-glutamyltransferase activities were measured. The CRF2R agonist increased extensor digitorum longus and soleus muscle weights (P < 0.05) in wildtype and mdx mice. Sedentary mdx CRF2R and exercised mdx placebo mice had lower serum creatine kinase activity than sedentary mdx placebo mice. CRF2R-treated mice had decreased heart weights compared to placebo-treated mice. We conclude that CRF2R agonists should be further evaluated as a potential therapy for dystrophinopathies.