Topochemical factors in potentiation of contraction by heavy metal cations

In addition to the previously studied Zn²⁺, low concentrations (about 0.5 mM) of Be²⁺, Ba²⁺, Cd²⁺, Ni²⁺, Cu²⁺, Pt⁴⁺ and, outstandingly, 0.5 µM of UO₂ ²⁺, potentiate the twitch of frog sartorius and toe muscles by prolonging the active state of contraction. The degree of potentiation is a roughly S-shaped function of p(metal²⁺), suggesting that each metal binds to a ligand of the muscle fiber, representative apparent affinity constants being: UO₂ ²⁺, 5 x 10⁶; Zn²⁺, 2.8 x 10⁵; and Cd²⁺, 2 x 10⁴. UO₂ ²⁺ potentiation effects are rapidly reversed by PO₄, and Zn²⁺ and Cd²⁺ effects by EDTA, PO₄, and cysteine. The rapidity of these reversals by the nonpenetrating EDTA and PO₄, and the fact that heavy metal ions evidently potentiate by prolonging the action potential, indicate that the metal potentiators exert their primary action at readily accessible (i.e. plasma and T tubular) membrane sites. The relatively slow kinetics of development of potentiation, and the even slower reversal of it in pure Ringer''s solution, indicate that the metal ions are bound to connective tissue, as well as to muscle fibers. The binding effects at the readily accessible membrane sites evidently impairs delayed rectification and thus modifies the action potential and excitation-contraction coupling so as to cause potentiation. SH is excluded, and PO₄ and imidazole are possibilities, as the membrane ligand binding the potentiating metal ions.     

Torque potentiation and myosin light-chain phosphorylation in human muscle following a fatiguing contraction

Indices of electrically stimulated and maximal voluntary isometric muscle torgue and the phosphate content of myosin phosphorylatable light chains (P light chains) were studied during recovery following a 60-s maximal voluntary isometric contraction (MVC) in 21 human subjects. Analysis of muscle biopsy samples revealed that immediately after the 60-s MVC there were significant decreases in ATP (-15%) and phosphocreatine (-82%), and lactate concentration increased by 17-fold. All indices of muscle torque production were reduced by the 60-s MVC, but the twitch torque and torque at 10 Hz were relatively less reduced compared with the torque at 20 and 50 Hz or a 1-s MVC. Between 3 and 6 min of recovery, twitch torque and torque at 10 Hz stimulation were significantly potentiated, reaching peak values of 125 and 134%, respectively, compared with rest. Phosphate content of the fast and two slow P light chains was significantly increased over rest levels immediately after and 4 min after the 60-s MVC. These results suggest that myosin P light-chain phosphorylation could provide a mechanism to increase human muscle torque under conditions of submaximal contractile element activation following fatigue.     

Mechanism of potentiation by diazepam of adenosine response

Diazepam (3 microM) potentiates the adenosine-induced relaxation of caecum. Other benzodiazepine receptor ligands, such as inosine and hypoxanthine, failed to modify adenosine responses. Diazepam failed to further modify the dose-response curve to adenosine obtained in the presence of dipyridamole, and uptake inhibitor. Diazepam and dipyridamole did not affect the responses to 2-chloroadenosine. The relaxant effect of adenosine was not blocked by Ro 15-1788, a benzodiazepine receptor antagonist. These observations indicated that diazepam potentiated adenosine response by inhibiting the uptake of purine nucleotide and, that the benzodiazepine receptor is not purinergic in nature in the rat caecum.     

Postactivation potentiation, fiber type, and twitch contraction time in human knee extensor muscles.

In small mammals, muscles with shorter twitch contraction times and a predominance of fast-twitch, type II fibers exhibit greater posttetanic twitch force potentiation than muscles with longer twitch contraction times and a predominance of slow-twitch, type I fibers. In humans, the correlation between potentiation and fiber-type distribution has not been found consistently. In the present study, postactivation potentiation (PAP) was induced in the knee extensors of 20 young men by a 10-s maximum voluntary isometric contraction (MVC). Maximal twitch contractions of the knee extensors were evoked before and after the MVC. A negative correlation (r = -0. 73, P < 0.001) was found between PAP and pre-MVC twitch time to peak torque (TPT). The four men with the highest (HPAP, 104 +/- 11%) and lowest (LPAP, 43 +/- 7%) PAP values (P < 0.0001) underwent needle biopsies of vastus lateralis. HPAP had a greater percentage of type II fibers (72 +/- 9 vs. 39 +/- 7%, P < 0.001) and shorter pre-MVC twitch TPT (61 +/- 12 vs. 86 +/- 7 ms, P < 0.05) than LPAP. These data indicate that, similar to the muscles of small mammals, human muscles with shorter twitch contraction times and a higher percentage of type II fibers exhibit greater PAP.     

Mechanism of apoptosis induced by IFN-alpha in human myeloma cells: role of Jak1 and Bim and potentiation by rapamycin

Interferon-alpha (IFN-alpha) has been used for the last 20 years in the maintenance therapy of multiple myeloma (MM), though it is only effective in some patients. Congruent with this, IFN-alpha induces apoptosis in some MM cell lines. Understanding the mechanism of IFN-alpha-induced apoptosis could be useful in establishing criteria of eligibility for therapy. Here we show that IFN-alpha-induced apoptosis in the MM cell lines U266 and H929 was completely blocked by a specific inhibitor of Jak1. The mTOR inhibitor rapamycin mitigated apoptosis in U266 but potentiated it in H929 cells. IFN-alpha induced PS exposure, DeltaPsi(m) loss and pro-apoptotic conformational changes of Bak, but not of Bax, and was fully prevented by Mcl-1 overexpression in U266 cells. IFN-alpha treatment caused the release of cytochrome c from mitochondria to cytosol and consequently, a limited proteolytic processing of caspases. Apoptosis induced by IFN-alpha was only slightly prevented by caspase inhibitors. Levels of the BH3-only proteins PUMA and Bim increased during IFN-alpha treatment. Bim increase and apoptosis was prevented by transfection with the siRNA for Bim. PUMA-siRNA transfection reduced electroporation-induced apoptosis but had no effect on apoptosis triggered by IFN-alpha. The potentiating effect of rapamycin on apoptosis in H929 cells was associated to an increase in basal and IFN-alpha-induced Bim levels. Our results indicate that IFN-alpha causes apoptosis in myeloma cells through a moderate triggering of the mitochondrial route initiated by Bim and that mTOR inhibitors may be useful in IFN-alpha maintenance therapy of certain MM patients.     

Mechanism of TrkB-mediated hippocampal long-term potentiation

The TrkB receptor tyrosine kinase and its ligand, BDNF, have an essential role in certain forms of synaptic plasticity. However, the downstream pathways required to mediate these functions are unknown. We have studied mice with a targeted mutation in either the Shc or the phospholipase Cgamma (PLCgamma) docking sites of TrkB (trkB(SHC/SHC) and trkB(PLC/PLC) mice). We found that hippocampal long-term potentiation was impaired in trkB(PLC/PLC) mice, but not trkB(SHC/SHC) mice. BDNF stimulation of primary neurons derived from trkB(PLC/PLC) mice fully retained their ability to activate MAP kinases, whereas induction of CREB and CaMKIV phosphorylation was strongly impaired. The opposite effect was observed in trkB(SHC/SHC) neurons, suggesting that MAPKs and CREB act in parallel pathways. Our results provide genetic evidence that TrkB mediates hippocampal plasticity via recruitment of PLCgamma, and by subsequent phosphorylation of CaMKIV and CREB.     

Effect of conditioning contraction intensity on postactivation potentiation is muscle dependent

We aimed to examine whether the influence of conditioning contraction intensity on the extent of postactivation potentiation (PAP) is muscle dependent. Eleven healthy males performed both thumb adduction and plantar flexion as a conditioning contraction. The conditioning contraction intensities were set at 20%, 40%, 60%, 80%, or 100% of the maximal voluntary isometric contraction (MVC).Before and after the conditioning contraction, twitch torque was measured for the respective joint to calculate the extent of PAP. In plantar flexion, the extent of PAP became significantly larger as the conditioning contraction intensity increased up to 80% MVC (p < 0.05). In contrast, the extent of PAP in thumb adduction increased significantly only up to 60% MVC (p < 0.05), but not at higher intensities.These results indicate that the influence of the conditioning contraction intensity on the extent of PAP is muscle dependent. Our results suggest that a conditioning contraction with submaximal intensity can sufficiently evoke sizable PAP in the muscle where most of muscle fibers are recruited at submaximal intensities, thereby attenuating muscle fatigue induced by the conditioning contraction.     

Evaluation of human dynamic contraction by phonomyography.

Phonomyogram (PMG, or acoustic myogram) is known to increase with force in isometric contractions. We investigated this relationship for dynamic contractions against different inertias. PMG and surface electromyogram (EMG) from biceps brachii and brachioradialis muscles were simultaneously recorded with the angular acceleration of elbow flexions. These were self-initiated movements (30 degrees) toward a fixed target and performed against two different inertias. PMG and EMG were integrated from the onset of the signal to the end of the acceleration phase. Phono- and electromechanical delays were also measured. For integrated EMG (iEMG), there was a linear relationship between integrated PMG (iPMG) and force, the slope of which did not depend on inertia. There was also a linear relationship between iPMG or iEMG and angular acceleration, with a higher slope for the highest inertia condition. There was also a family of linear relationships between iPMG or iEMG and angular acceleration, and their slopes depended on inertia. Measurements of the phono- and electromechanical delays showed that onset of PMG followed that of EMG but preceded onset of acceleration. It is suggested that PMG expresses tension of the underlying muscle contractile elements. Given the simplicity of the PMG method, we conclude that PMG allows convenient evaluation of muscle tension during human dynamic contraction.     

Posttetanic potentiation of human dorsiflexors

O'Leary, Deborah D., Karen Hope, and Digby G. Sale.Posttetanic potentiation of human dorsiflexors.J. Appl. Physiol. 83(6):2131-2138, 1997.Twitch contractions of the ankle dorsiflexors were evoked before and after applied 7-s tetanic stimulation at 100 Hzin 20 young adults. Torque decreased 15% during the tetanus. At 5 safter tetanus, twitch peak torque had potentiated 45%. Potentiationdeclined to 28% after 1 min, rose slightly to 33% at 2 min, anddeclined slowly with potentiation still 25% after 5 min. There waslarge intersubject variation in the amount of potentiation(5-140%) and its persistence (5 to 20 min). The muscle compoundaction potential (M wave) did not change significantly (from pretetanicvalue) at 5 s after tetanus but increased sharply (26%) at 2 min andthen subsided. Twitch half relaxation time (23%) decreasedsignificantly more than twitch rise time (13%) 5 s after tetanus andrecovered more slowly. Twitch rates of torque development (75%) andrelaxation (71%) increased similarly 5 s after tetanus and were stillelevated (~25%) at 5 min. The extent of twitch torque potentiationwas significantly inversely correlated with pretetanic twitch rise time(r = 0.69), half relaxation time (r = 0.61), andtwitch-to-tetanus ratio (r = 0.66). The data indicate that posttetanic potentiation has agreater effect on twitch half relaxation time than on time to peaktorque and is more prominent in muscles with a short twitch time courseand small twitch-to-tetanus ratio.

     

Effect of different types of conditioning contraction on upper body postactivation potentiation

Muscle contractions preceding an activity can result in increased force generation (postactivation potentiation [PAP]). Although the type of muscular contractions could affect subsequent strength and power performance, little information exists on their effects. The purpose of this study was to examine PAP effects produced by isometric (ISO), concentric (CON), eccentric (ECC), or concentric-eccentric (DYN) conditioning contractions on upper body force and power performance. Ten male, competitive rugby players (mean ± SD: age 20.4 ± 0.8 years, height 177.0 ± 8.1 cm, body mass 90.2 ± 13.8 kg) performed a ballistic bench press throw (BBPT) followed by a 10-minute rest and one of the conditioning contractions. After a 12-minute rest, the subjects performed another BBPT (post-BBPT). The conditioning contractions, applied on separate days and in counterbalanced randomized order, were a 7-second isometric barbell bench press for ISO and 1 set of 3 bench press repetitions at 3 repetition maximum for CON, ECC, and DYN (each repetition lasting 2 seconds for CON and ECC, overall execution time <7 seconds for DYN). Peak power (Ppeak), peak force (Fpeak), maximum distance (Dmax) and rate of force development (RFD) were measured using a linear position transducer. Electromyography (EMG) of the pectoralis major and triceps brachii was also recorded. The ISO produced significantly higher Ppeak (587 ± 116 and 605 ± 126 W for pre- and post-BBPT, respectively; p < 0.05). No significant differences in Ppeak were revealed for CON, ECC, and DYN (p > 0.05), and no significant differences existed in Fpeak, Dmax, and RFD for ISO, CON, ECC, and DYN (p > 0.05). Finally, EMG was not significantly different between pre- and post-BBPT for any of the conditioning contractions (p > 0.05). Isometric contractions appear to be the only conditioning contractions increasing upper body power output after long resting periods.     

Mechanism of long-term potentiation of transmitter release induced by adrenaline in bullfrog sympathetic ganglia

A mechanism of the long-term potentiation of transmitter release induced by adrenaline (ALTP) was studied by recording intracellularly the fast excitatory postsynaptic potentials (fast EPSPs). The ALTP was produced during the blockade of K+ channels at the presynaptic terminals by tetraethylammonium (TEA). The synaptic delay, possibly reflecting a relative change in the duration of an action potential at the presynaptic terminal, was not changed during the course of the ALTP. By contrast, it was significantly lengthened by TEA and other K+ channel inhibitors (4-aminopyridine and Cs+) that markedly enhanced the evoked release of transmitter. The magnitude of facilitation of the fast EPSP, induced by a conditional stimulus to the preganglionic nerve, was decreased during the generation of the ALTP, but was unchanged during the potentiation of transmitter release caused by TEA. These results, together with theoretical considerations applying the residual Ca2+ hypothesis to the facilitation, suggest that the enhancement of transmitter release during the ALTP is not caused by an increased Ca2+ influx during a presynaptic impulse owing to the blockade of K+ channel or the modulation of Ca2+ channel, but presumably is induced by a rise in the basal level of free Ca2+ in the presynaptic terminal.     

High-throughput profiling of dietary polyphenols and their metabolites by HPLC-ESI-MS-MS in human urine

Polyphenols in biological fluids are generally estimated by HPLC with UV, electrochemical or fluorimetric detection. We describe herein a method specially developed to estimate biomarkers of polyphenol consumption in human urine. We simultaneously quantified 15 polyphenols and their related compounds in human urine using an HPLC coupled with electrospray ionization mass-mass (HPLC-ESI-MS-MS) method with an analytical run time of 6 min. The method has been validated with respect to linearity, precision, and accuracy in intra- and inter-day assays. It has been applied to human urine samples collected from volunteers after consumption of different polyphenol-rich beverages in controlled conditions.     

Simultaneous potentiation and fatigue in quadriceps after a 60-second maximal voluntary isometric contraction

Potential mechanisms of fatigue (metabolic factors) and potentiation (phosphate incorporation by myosin phosphorylatable light chains) were investigated during recovery from a 60-s maximal voluntary isometric contraction (MVC) in the quadriceps muscle of 12 subjects. On separate days before and for 2 h after the 60-s MVC, either a 1-s MVC or electrically stimulated contractions were used as indexes to test muscle performance. Torque at the end of the 60-s MVC was 57% of the initial level, whereas torques from a 1-s MVC and 50-Hz stimulation were most depressed in the immediate recovery period. At this time, muscle biopsy analyses revealed significant decreases in ATP and phosphocreatine and a 19-fold increase in muscle lactate. Conversely, isometric twitch torque and torque from a 10-Hz stimulus were the least depressed of six contractile indexes and demonstrated potentiation of 25 and 34%, respectively, by 4 min of recovery (P less than 0.05). At this time, muscle lactate concentration was still 16 times greater than at rest. An increased phosphate content of the myosin phosphorylatable light chains (P less than 0.05) was also evident both immediately and 4 min after the 60-s MVC. We conclude that the 60-s MVC produced marked force decreases likely due to metabolic displacement, while the limited decline in the twitch and 10-Hz torques and their significant potentiation suggested that myosin phosphorylation may provide a mechanism to enhance contractile force under conditions of submaximal activation during fatigue.     

Dietary polyphenols decrease glucose uptake by human intestinal Caco-2 cells

The effect of different classes of dietary polyphenols on intestinal glucose uptake was investigated using polarised Caco-2 intestinal cells. Glucose uptake into cells under sodium-dependent conditions was inhibited by flavonoid glycosides and non-glycosylated polyphenols whereas aglycones and phenolic acids were without effect. Under sodium-free conditions, aglycones and non-glycosylated polyphenols inhibited glucose uptake whereas glycosides and phenolic acids were ineffective. These data suggest that aglycones inhibit facilitated glucose uptake whereas glycosides inhibit the active transport of glucose. The non-glycosylated dietary polyphenols appear to exert their effects via steric hindrance, and (-)-epigallochatechingallate, (-)-epichatechingallate and (-)-epigallochatechin are effective against both transporters.