Efficiency of work performance and contraction velocity in isotonic tetani of frog sartorius

Mechanical work, ATP, ADP, PC, free creatine and lactate concentrations were determined on IAA poisoned frog sartorii tetanically stimulated in humidified N, at 10°C in isotonic conditions (0.25 or 0.45 P₀). Tetanus duration was 0.35 s, number of tetani was varied from 0 (rest) to 25 (exhaustion). The mechanical work performed per mole ATP+PC split (W _P ^* ) amounted on the average to 16.7 kJ/mol. It was observed, however, that w _p ^* increased from about 13 to about 24 kJ/mol with decreasing ATP concentration from about 2 (resting value) to about 1 mol/g and that this decrease in ATP was associated with a decrease of the shortening (and relaxation) speed of the muscle to about 30% of the values observed on the first tetanus. It is concluded that the thermodynamic efficiency of muscle contraction, calculated from the ratio of w _P ^* (measured) to the thermodynamic affinity (free energy change) of ATP hydrolysis (estimated) increases from about 0.3 to about 0.5 with decreasing ATP concentration and shortening speed.This work was supported by the Fonds National Suisse de la Recherche Scientifique, Grants 3.332.78 and 3.364.0.82     

Human T-lymphocyte subset production of immune (γ) interferon

Human T, T, and T lymphocyte subpopulations have the capacity to respond to phytohemagglutinin (PHA)in vitro with proliferation and the production of a pH 2 and heat-labile interferon. This occurs both when the subsets are isolated by direct rosetting techniques or by negative selection. Macrophages enhance the production of the interferon by each lymphocyte subset and do not themselves produce interferon in response to products of PHA-activated lymphocyte subsets. Thus our studies indicate that subpopulations of T lymphocytes known to differ with regard to morphology, surface receptors, RNA content, response to corticosteroids and X-irradiation, and other functional capabilities do not differ with regard to their capacity to produce interferon.     

Chemical energy usage during isometric twitches of frog sartorius muscle intoxicated with an isomer of creatine,β-guanidinopropionate

Frogs were injected for several weeks with -guanidinopropionate, an isomer of creatine. Their sartorius muscles were isolated, poisoned with iodoacetate and stimulated isometrically with 75 shocks/min in nitrogen until rigor. In comparison with sartorius muscles of untreated frogs, they contained more free creatine and less phosphocreatine, but the same content in total creatine and ATP. They also contained -guanidinopropionate both free and phosphorylated. However, muscles in rigor contained the same concentration of the phosphorylated form as resting muscles, i.e., phospho--guanidinopropionate was not split during contraction. The number of twitches performed before rigor was decreased. There was no change in the chemical energy usage (sum of phosphocreatine breakdown and twice ATP breakdown) per twitch.     

Energetics of anaerobic glycolysis in dog gastrocnemius

Thermally isolated gastrocnemii were stimulated to exhaustion, by rhythmic isotonic (70 N) tetanic contractions, during complete occlusion of blood flow. Enthalpy change (h=work + heat) and work output (w) (kJ/kg) were obtained from records of deep muscle temperature and shortening. The lactate produced (LA, mol/kg) was measured in the outflow after reestablishement of blood flow. The following relationships were obtained:h=76 LA+1.2, andw=19.8 LA+0.30. As the energy liberated at exhaustion by alactic energy sources (P and O₂ stores) is constant, h/ LA=76 (±10.5, S.E.) kJ/mol is the enthalpy change for lactate formation ( ). The neutralization heat was estimated on muscle homogenates at 12 kJ/mol, leaving 64 kJ/mol for of LA formation proper. The mechanical efficiencies of work related to LA formation (E _LA) and of that not related to LA formation (E _nonLA) were practically identical (0.25). From these values and from , the enthalpy change of P splitting was estimated in the range 52–62 kJ/mol, depending on the value of the ratio P/ LA assumed in the calculation.     

Effects of aging on the mechanical threshold of rat skeletal muscle fibers

Mechanical threshold was measured in vitro in extensor digitorum longus (EDL) muscle fibers from rats of 3–4 and 29 months of age, by means of a two microelectrode point voltage clamp. The potential needed for evoking a barly visible contraction was determined using depolarizing command pulses of 5–500ms duration. At each pulse duration, the EDL fibers from aged rats contracted at a significantly more negative potential than did those from the younger adult rats. Accordingly, the strength duration curve of the aged EDL was significantly shifted towards more negative potentials compared to that for adult rats. The rheobase voltages estimated from the fit of such curves were –62.6±0.81mV and –57.1±0.87mV in aged ana adult EDL fibers, respectively. The data suggest that changes in excitation-contraction coupling parallel the prolongation of contractile times observed during aging in mammalian skeletal muscle. These results are consistent with the known reduction in rate and extent of Ca⁺⁺ uptake by sarcoplasmic reticulum in aged rats.     

Cow milk angiogenin induces cytokine production in human blood leukocytes

Angiogenin isolated from cow milk induces the production of cytokines IL-1, IL-6, and TNF- in human leukocytes; the level of production of each cytokine depends on the concentration of the preparation, and the dynamics of production depends on the time from the beginning of induction. Simultaneous treatment with angiogenin and phytohemagglutinin had an additive effect on the production of cytokines, the time of this effect manifestation being individual for each cytokine.     

Glucocorticoids modulate TGF-beta production

TGF- is thought to play a central role in pulmonary fibrosis inducing fibroblast differentiation and extracellular matrix synthesis. In human lung fibroblasts, it is still unclear how various TGB- isoforms affect TGF- production and whether glucocorticoids, commonly used agents to treat fibrotic lung disease, modulate these processes. To this end, human fetal lung fibroblasts (HFL-1) were cultured with various concentrations of glucocorticoids (budesonide, dexamethasone or hydrocortisone) with and without TFG-1, -2, and -3. TGF- mRNA was assessed by real time RT-PCR. Smad 2, 3, and 4 and AP-1 complex (c-fos and c-Jun) cellular localization were evaluated by immunostaining. TGF-2 and -3 stimulated TGF-1 production significantly (p < 0.01 relative to control). TGF-1 stimulated TGF-2 production (p < 0.01 relative to control). TGF-3 was undetectable. Glucocorticoids significantly inhibited TGF-1 and -2 production and reduced expression of the upregulated TGF-1 and -2 mRNA induced by exogenous TGF-1, -2 or -3 (p < 0.01 for each) but had no effect on Smads. Although c-jun-related nuclear staining was not intensified in TGF--stimulated cells, it was reduced by glucocorticoids. Thus, TGF- isoforms may stimulate production of various TGF- isoforms in the lung. Glucocorticoids then may block TGF- production by modulating mRNA levels and c-Jun.     

Human arm stiffness characteristics during the maintenance of posture

Summary When the hand is displaced from an equilibrium position, the muscles generate elastic forces to restore the original posture. In a previous study, Mussa-Ivaldi et al. (1985) have measured and characterized the field of elastic forces associated with hand posture in the horizontal plane. Hand stiffness which describes the relation between force and displacement vectors in the vicinity of equilibrium position was measured and graphically represented by an ellipse, characterized by its size, shape and orientation. The results indicated that the shape and orientation of the stiffness ellipse are strongly dependent on arm configuration. At any given hand position, however, the values of these parameters were found to remain invariant among subjects and over time. In this study we investigate the underlying causes for the observed spatial pattern of variation of the hand stiffness ellipse. Mathematically analyzing the relation between hand and joint stiffness matrices, we found that in order to produce the observed spatial variations of the stiffness ellipse, the shoulder stiffness must covary in the workspace with the stiffness component provided by the two-joint muscles. This condition was found to be satisfied by the measured joint stiffness components. Using anatomical data and considering the effects that muscle cross-sections and changes in muscle moment arms have on the joint stiffness matrix, we found that these anatomical factors are not sufficient to account for the observed pattern of variation of joint stiffness in the workspace. To examine whether the coupling between shoulder and two-joint stiffnesses results from the coactivation of muscles contributing to these stiffnesses, EMG signals were recorded from shoulder, elbow and two-joint muscles. Our results indicated that, while some muscle coactivation may indeed exist, it can be found for only some of the muscles and in only part of the workspace.     

An analysis of postsynaptic potentials of tectal neurons of the frog: correlation with impulses recorded from the terminals of retinotectal afferents

Summary The purpose of this study is to examine the synaptic action between terminals of retinal ganglion cell axons and tectal neurons. To accomplish this, an extracellular single unit identified as retinotectal fiber was first isolated from the superficial layer of the optic tectum and intracellular responses were recorded from a tectal neuron in the vicinity of the extracellular recording electrode. On-off retinal fibers and both E-E (EPSP at on and off of diffuse light) and EI-EI type (EPSP-IPSP combination at on and off of diffuse light) tectal neurons were selected for the pre- and postsynaptic pair. Postsynaptic responses to a small moving square were averaged by triggering with the isolated presynaptic impulses. The latency of the resultant EPSPs indicated that most of the E-E and EI-EI type tectal neurons were monosynaptically activated by on-off retinal fibers. One of the E-E type tectal neurons was identified as a large ganglionic neuron in layer 8.     

Reduced intramembrane charge movement in the dysgenic skeletal muscle cell

Intramembrane charge movement in skeletal muscle cells has been proposed to underlie the process leading to Ca release from the sarcoplasmic reticulum. A number of recent studies suggest that the dihydropyridine receptor located in the transverse-tubular membrane is responsible for the generation of intramembrane charge movement. The skeletal muscle cell of the mutant mouse with Muscular Dysgenesis is characterized by absence of excitation-contraction coupling. Here we investigated the charge movement in freshly dissociated skeletal muscle cells from dysgenic mice. In 9 out of 34 dysgenic mouse cells the charge movement was completely absent, in the remaining cells the charge movement was never more than 30 % of control. The amount of maximum charge movement (Q_max) in mutant muscle cells was less than 30 % of Q_max in normal muscle. Nifedipine, a dihydropyridine derivative, reduced the amount of charge movement in normal muscle cells but it was less effective on charge movement in mutant muscle cells. We conclude that there is an alteration of nifedipine-sensitive charge movement in the skeletal muscle cells from the mutant mice.     

Skeletal muscle: Dependence of potassium contractures on extracellular calcium

Summary The effects of Ca-free salines were tested in frog skeletal muscle fibres. It was found that the decrease of the (Ca) in the saline from 1.8 mM to about 3.2×10^–10 M (with 4 mM Mg present) did not change the resting potential of the fibres and the depolarizations induced by 40 and 80 mM K. The action potentials, however, were significantly reduced in amplitude and duration by 24% and 33% respectively. K contractures induced with a 117 mM KCl saline had a peak and a slow component. The exposure of the fibres to a Ca-free saline [(Ca)ca. 1.6×10^–9M] for 40 sec reduced the slow component to 39±16.5% (mean±s.e; 6 fibres) of the control value. The peak component, on the other hand, was slightly decreased to 78±10.5%. These effects were reversible.The inhibition of the K contractures by extremely low (Ca)_o cannot be explained by an alteration of the electrical properties of the muscle fibres or by a depletion of the Ca content of the sarcoplasmic reticulum. It is postulated that the drastic removal of external Ca reduces the driving force for a Ca influx that may take place during the excitation of the muscle and that may be essential for the excitation-contraction coupling.This work was supported by grants from the Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina (CONICET) and, partially, from the USPHS (ROI-NS 06935-03NEUA).     

Synoviocytes in chronic synovitis in situ and cytokine stimulated synovial cells in vitro neo-express α1, α3 and α5 chains of β1 integrins

The expression of the 1 integrins was examined immunohistochemically in synoviocytes from normal synovial membrane and from chronic synovitis of different aetiology and intensity. Normal synoviocytes were 61-positive but lacked 1 through 5. In mild inflammation type A synoviocytes neo-expressed 1, 3, and 5 chains. In severe inflammation both type A and B synoviocytes expressed 3, 4, 5, and 6 chains. The effects of inflammatory cytokines, as single agents or in combination, on the 1 integrin expression in cultured normal synoviocytes was determined by immunocytochemistry and flow cytometry. The 1 chain, while absent in unstimulated synoviocytes, was induced by interleukin-1 (IL-1), tumour necrosis factor- (TNF-), and interferon- (INF-). This effect was enhanced by combining IL-1 and TNF-. Expression of the 3 chain was up-regulated by IL-1 and, more intensely, by IFN-. Transforming growth factor (TGF-) inhibited the up-regulating effect of IL-1 and antagonized the effect of IFN- on 3 chain expression. Expression of the 5 chain was up-regulated significantly by co-stimulation through IL-1 together with TGF- or TNF-. Thus, the 1 integrin profile of cytokine activated synoviocytes in vitro resembled that of synoviocytes in synovitis in situ. These data suggest that IL-1, TNF-, IFN-, and TGF- are likely to be among the effectors regulating 1 integrin expression in synoviocytes in vivo.     

The isolated frog skin epithelium: Presence of α and β adrenergic receptors regulating active sodium transport and water permeability

Summary The effects of stimulation of and adrenergic receptors on short circuit current (S.C.C.), Na⁺ and Cl^– fluxes and osmotic water permeability were studied on isolated frog skin epithelial layers separated from the dermis.Low norepinephrine doses (final concentrations in the incubation medium ranging from 5×10^–9 to 10^–8 M) produced increased water permeability and S.C.C. The latter was entirely accounted for by an increase in the active Na⁺ influx. Na⁺ outflux and Cl^– fluxes were not modified. Both these effects disappeared after treatment with the blocking agent, Propranolol. Higher norepinephrine doses (final concentrations: 10^–7 to 10^–6 M) produced: 1. an increase in water permeability lower than that produced by low doses, the highest doses failing to increase water permeability, and 2. a triphasic change in S.C.C.: after an initial increase, S.C.C. dropped to its resting value and then rose again to a sustained value. Na⁺ and Cl^– flux measurements showed that the variation in S.C.C. reflected variations in active Na⁺ transport. When the same high norepinephrine doses were applied after treatment with the blocking agent Phentolamine, the effects observed were identical to those obtained with low doses.On blocked preparations, large doses of norepinephrine inhibited the water permeability and sodium transport increases induced by theophylline or oxytocin but did not modify those induced by 35-cyclic AMP. The inhibition was suppressed after blocking receptors.From the foregoing, it was concluded that both and adrenergic receptors are present in frog skin epithelial cells and are involved in the regulation of water and sodium permeability.It is suggested that the inhibitory effect of stimulation resulted from the inhibition of cyclic-AMP generating system, the activity of which is under the positive control effect of oxytocin and stimulation.     

Statin treatment and a disease-specific pattern of β-amyloid peptides in Alzheimer’s disease

According to the amyloid cascade hypothesis, sporadic Alzheimers disease (AD) is caused by the production and aggregation of -amyloid (A), and the production of A has recently been linked to the metabolism of cholesterol. We have previously published clinical studies where the effect of statin treatment on A production has been investigated. No effect on A was found, which is in disagreement with cell and animal studies. In the present study we investigated the effect of statin treatment on a disease-specific pattern consisting of a C-terminally-truncated quintet of A peptides. Nineteen patients with AD were treated with simvastatin for 12 months and the quintet of A peptides were analysed in cerebrospinal fluid before and after treatment. Also included was a group of 15 untreated patients with AD. We found that the A peptide pattern at baseline was in agreement with earlier findings; however, we did not find any change in the A peptide pattern after statin treatment. We suggest that clinical studies with extended treatment periods are performed where higher dosages of statins are used. We also believe that the pleiotropic effects of statins should be investigated further in order to elucidate the connection between Alzheimers disease and statin treatment.