Therapeutic electrical stimulation and immune status in healthy men.

In physical and rehabilitation medicine, there are few reports on the effects of therapeutic low-frequency electrical stimulation on the immune response of the organism, even though electrical stimulation is used widely in clinical practice and sports medicine. The aim of our study was to examine the possible immunological consequences of moderate transcutaneous neuromuscular electrical stimulation (NMES) for quadriceps muscle strengthening in healthy individuals. The study included twelve healthy male adult volunteers (mean age 42 years) without contraindications for electrical stimulation. At the beginning and immediately after a 20-min session of NMES of quadriceps muscles, peripheral blood was collected to analyse the biochemical blood components (creatinine, creatine kinase, estimated glomerular filtration rate, cortisol), differential white blood cell count and immunological parameters. The intensity of NMES was set at maximum tolerance, eliciting on average about one-sixth of the maximum voluntary isometric contraction of the same leg. No statistically significant differences in the average group level were found in any of the measured biochemical blood components, white blood cell count or immunological parameters after the NMES session. On an individual level, the changes in creatine kinase, estimated glomerular filtration rate, basophils and some immunological parameters correlated with changes in the cortisol level. We can conclude that moderate transcutaneous low-frequency electrical stimulation for quadriceps muscle strengthening used in our study did not induce essential changes in immune status in healthy men.

The dual CCR2/CCR5 chemokine receptor antagonist Cenicriviroc reduces macrophage infiltration and disease severity in Duchenne muscular dystrophy (Dmdmdx-4Cv) mice.

Duchenne muscular dystrophy (DMD) is characterized by progressive muscle weakness which is ultimately fatal, most often due to involvement of the diaphragm. Macrophage infiltration of dystrophic muscles has been strongly linked to muscle damage and fibrosis in DMD. We hypothesized that cenicriviroc (CVC), a dual chemokine receptor (CCR2/CCR5) antagonist currently under clinical evaluation for other diseases, could prevent macrophage accumulation and blunt disease progression in the diaphragms of mdx mice (genetic homologue of DMD). Treatment with CVC (20 mg/kg/day intraperitoneally) or vehicle was initiated in mdx mice at 2 weeks of age (prior to the onset of muscle necrosis) and continued for 4 weeks. Flow cytometry to assess inflammatory cell subsets as well as histological and force generation parameters were determined in mdx diaphragms at the conclusion of the treatment. CVC therapy induced a major (3.9-fold) reduction in total infiltrating macrophages, whereas total numbers of neutrophils and T lymphocytes (CD4+ and CD8+) were unaffected. No changes in macrophage polarization status (inflammatory versus anti-inflammatory skewing based on iNOS and CD206 expression) were observed. Muscle fiber size and fibrosis were not altered by CVC, whereas a significant reduction in centrally nucleated fibers was found suggesting a decrease in prior necrosis-regeneration cycles. In addition, maximal isometric force production by the diaphragm was increased by CVC therapy. These results suggest that CVC or other chemokine receptor antagonists which reduce pathological macrophage infiltration may have the potential to slow disease progression in DMD.

The proinflammatory cytokines TNF-alpha and IL-1 beta impair economy of contraction in human myocardium.

Considerable experimental evidence has accumulated over the past years that proinflammatory cytokines, especially TNF-alpha and IL-1beta, impair myocardial function in different animal species. On the other hand, several prospective clinical trials studying TNF-alpha antagonist in patients with chronic heart failure were not able to demonstrate a benefit. As there might be a relevant species-related discrepancy, we intended to prove our previous results demonstrating impaired myocardial economy after exogenous administration of recombinant TNF-alpha in rat myocardium. In the present study, both TNF-alpha and IL-1beta not only revealed an immediate negative inotropic effect but also increased specific oxygen demand in human right-atrial myocardium. Enhanced oxygen consumption was not caused by an elevated basal metabolism but an impaired economy of contraction. Our results suggest that proinflammatory cytokines have a considerable effect on myocardial mechano-energetic parameters in human myocardium as well.

Roles of P2X receptors and Ca2+ sensitization in extracellular adenosine triphosphate-induced hyperresponsiveness in airway smooth muscle.

BACKGROUND: The release of adenosine triphosphate (ATP) from the airway epithelial cells during the inflammatory process is considered to play an important role in the pathophysiology of asthma and chronic obstructive pulmonary disease. OBJECTIVE: This study was designed to determine whether extracellular ATP is involved in the bronchial hyperresponsiveness as an interaction between epithelium and smooth muscle in the airways. METHODS: We examined the contractile response to methacholine (MCh) before and after exposure to low concentrations (< or = 10 microm) of ATP in isolated, epithelium-denuded guinea-pig tracheal smooth muscle by measuring isometric tension. Intracellular Ca2+ concentrations ([Ca2+]i) were assessed by fluorescent intensities of fura-2. RESULTS: MCh-induced contractile force was increased with no change in [Ca2+]i after exposure to 10 microm ATP for 15 min. The ability of ATP to enhance the MCh-induced contraction was markedly attenuated by suramin, a non-selective P2 receptor inhibitor. Pre-incubation with ATPgammaS, a non-hydrolysable analogue of ATP and alpha,beta-meATP, a P2X agonist, also enhanced the MCh-induced contraction. In contrast, uracil triphosphate, a P2Y agonist, did not affect the MCh-induced contraction. Y-27632, a Rho-kinase inhibitor, suppressed the ability of ATP to enhance the MCh-induced contraction. Moreover, PP1 and PP2, Src tyrosin kinase inhibitors, suppressed the enhancement of MCh-induced contraction by ATP. CONCLUSION: Pre-treatment with ATP induces hyperresponsiveness to MCh mediated by Ca2+ sensitization via the P2X receptor in airway smooth muscle. The present findings suggest the possible involvement of both the Rho-kinase and Src pathways in the intracellular mechanism of this phenomenon.

Essential role of myosin S-2 region in muscle contraction.

We studied the contraction characteristics and Mg-ATPase activity of glycerinated rabbit psoas muscle fibers in the presence and absence of polyclonal antibody directed against the subfragment-2 (S-2) region of myosin, to give information about the role of myosin hinge region in muscle contraction. The antibody was kindly supplied to us from Professor Harrington's laboratory. The antibody-induced decrease of Ca(2+)-activated isometric force development was always accompanied by a parallel decrease of muscle fiber stiffness, so that the stiffness versus force relation remained the same by the antibody treatment. Force-velocity curves, obtained by applying ramp decreases in load from steady isometric force to zero, indicated that the antibody had no effect on the maximum shortening velocity and the shape of the force-velocity curve. Simultaneous measurements of Mg-ATPase activity and Ca(2+)-activated isometric force showed that Mg-ATPase activity of the fibers remained unchanged despite the antibody-induced decrease of isometric force even to zero. These results indicate that, if the antibody attaches to the S-2 region of myosin molecules, their heads still hydrolyze ATP without contributing to both muscle force generation and muscle fiber stiffness.