The mechanism of cell entry and intracellular fate of a gene transfer vector composed of a receptor-targeting, DNA-condensing peptide, RGD-oligolysine, a luciferase encoding plasmid DNA (pDNA) and a cationic liposome was examined. We demonstrate by confocal microscopy, electron microscopy and subcellular fractionation that the major mechanism of entry of the vector is endocytic. The vector complex rapidly (5 min) internalizes into early endosomes, then late endosomes and lysosomes. Entry involves, at least in part, clathrin-coated pit-mediated endocytosis since different conditions or drugs known to influence this pathway modify both uptake of pDNA and its expression. The observed increase in expression with addition of a lip some

The relationships between force, electromyography (EMG), and muscle sounds recorded by acoustic myography (AMG) were investigated for both voluntary and stimulated isometric contractions in the adductor pollicis muscle. Voluntary activity was performed at 10, 25, 50, 75, 85, and 100% of maximal voluntary contraction (MVC) force. Stimulated contractions were produced by supramaximal electrical stimulation of the ulnar nerve at the wrist at frequencies of 10, 20, 30, 50, 70, and 100 Hz. Contractions lasted for 4 s each, and were performed in random order with a 3-min rest between each. The voluntary and stimulation studies were performed in random order between subjects. Simultaneous recordings were obtained for force, force oscillation (from the differentiated force signal), and raw and integrated AMG (IAMG) and EMG (IEMG). During voluntary contractions, IAMG increased with force up to MVC (r2 = 0.99, P less than 0.001) in a curvilinear fashion and a similar relationship was seen between force and IEMG (r2 = 0.99, P less than 0.001). Conversely, during stimulated contractions as stimulation frequency increased, IAMG decreased in a fashion mirroring the frequency-force curve. The frequency of the AMG signal matched stimulation frequency and declines in total IAMG were due to reductions in amplitude of the AMG signal. The stimulation frequency-oscillation of force relationship was identical to that seen for stimulation frequency and IAMG. Integrated EMG increased linearly with stimulation frequency (r = 0.99). The stimulation results suggest that muscle sounds reflect oscillation of muscle fibers and that AMG signal characteristics are determined by motor control mechanisms rather than intrinsic contractile processes.

Nonviral gene delivery systems consist predominantly ofExpression was further enhanced 30-fold (P Ͻ 0.01) with lipoplexes or receptor-targeting and nontargeting polylipofectamine and the ratio of DNA/peptide/lipofectamine plexes. We examined integrin-mediated gene delivery was critical for specificity and expression. Fluorescence using an Arg-Gly-Asp/oligo-L-lysine ([K]

Interleukin-6 (IL-6) concentrations in knee joint synovial fluids and paired plasma samples of arthritis patients were examined with respect to each other and parameters of the inflammatory response. Synovial fluid and plasma IL-6 concentrations were significantly higher in patients with inflammatory arthritis than those detected in patients with osteoarthritis (P less than 0.001). The IL-6 concentrations in synovial fluids were considerably higher than, but significantly correlated with (r = 0.65; P less than 0.001), those of plasma. Furthermore, synovial fluid IL-6 concentrations in bilaterally inflamed knees were significantly correlated (r = 0.79; P less than 0.001) and there was a significant correlation with the extent of inflammatory cell infiltrate (r = 0.75; P less than 0.001). In unselected rheumatoid arthritis patients there was only a weak correlation between IL-6 and plasma C-reactive protein (CRP) concentration, and no correlation between IL-6 and erythrocyte sedimentation rate (ESR). However, both ESR and CRP concentration were highly correlated with plasma IL-6 concentration in patients with other inflammatory arthritides, particularly psoriatic and HLA B27 positive spondyloarthritis (r = 0.72-0.94; P less than 0.005). These relationships suggest that IL-6 production in inflammed knee joints can be a significant determinant of acute phase protein responses in arthritis patients, although the situation in patients with rheumatoid arthritis is more complex and may be influenced by other disease-related factors.