Effect of laterotrusive occlusal scheme on chewing duration, external intercostal muscular activity, heart rate, and oxygen saturation.

Objective: To evaluate the effect of the laterotrusive occlusal scheme on chewing duration, external intercostal (EIC) electromyographic (EMG) activity, heart rate (HR), and oxygen saturation (OS) during different tasks in the upright seated position.Methods: Fifty young participants, 25 with canine guidance and 25 with group function, were included. Chewing duration, bilateral EIC EMG activity, HR, and OS were recorded during the following tasks: 1) chewing until swallowing threshold; 2) laterotrusive teeth grinding.Results: Chewing duration, bilateral EIC EMG activity, HR, and OS showed no significant differences between the two laterotrusive occlusal schemes during the tasks studied.Conclusion: These results suggest that chewing duration, EIC muscle activity, HR, and OS were not significantly influenced by the laterotrusive occlusal scheme. Therefore, when a modification of the laterotrusive occlusal scheme is needed during oral rehabilitation or orthodontic treatment, canine guidance or group function should not significantly change EMG activity of EIC muscles.

Awake teeth grinding in participants with canine guidance or group function: Effect on diaphragm EMG activity, heart rate, and oxygen saturation.

Objective: To compare the effect of canine guidance or group function on diaphragm activity, heart rate, and oxygen saturation during awake teeth grinding at different body positions. Methods: Fifty healthy participants, 25 with canine guidance and 25 with group function, were included. Bilateral electromyographic (EMG) recordings of the diaphragm (DIA) during awake teeth grinding were performed in standing, seated upright, and right lateral decubitus positions. Simultaneously, heart rate and oxygen saturation were measured. Results: EMG activity of the DIA muscle was similar in the working side and non-working side between participants with canine guidance and group function in the different body positions studied. The heart rate and oxygen saturation showed no significant differences between the two groups. Conclusion: EMG activity of the DIA muscle, the heart rate, and oxygen saturation during teeth grinding are not significantly influenced by the type of laterotrusive occlusal scheme.

Effect of breathing type on electromyographic activity of respiratory muscles during tooth clenching at different decubitus positions.

OBJECTIVE: To compare the effect of breathing type on electromyographic (EMG) activity of respiratory muscles during tooth clenching at different decubitus positions. METHODS: Forty young men participants were included, 11 with upper costal, 9 with mixed, and 20 with costo-diaphragmatic breathing type. EMG recordings of diaphragm (DIA), external intercostal (EIC), sternocleidomastoid (SCM), and latissimus dorsi (LAT) muscles during tooth clenching in the intercuspal position were performed in dorsal, left lateral, and ventral decubitus positions. RESULTS: DIA EMG activity was higher in subjects with upper costal or mixed than with costodiaphragmatic breathing type (p = 0.006; 0.021, respectively), whereas it was similar between upper costal and mixed breathing types. EIC, SCM, and LAT activity was similar among breathing types. CONCLUSION: Higher DIA activity would be a risk factor to exceed the adaptive capability of healthy subjects with upper costal or mixed breathing type.

N-Acetylcysteine Prevents the Spatial Memory Deficits and the Redox-Dependent RyR2 Decrease Displayed by an Alzheimer's Disease Rat Model.

We have previously reported that primary hippocampal neurons exposed to synaptotoxic amyloid beta oligomers (AßOs), which are likely causative agents of Alzheimer's disease (AD), exhibit abnormal Ca2+ signals, mitochondrial dysfunction and defective structural plasticity. Additionally, AßOs-exposed neurons exhibit a decrease in the protein content of type-2 ryanodine receptor (RyR2) Ca2+ channels, which exert critical roles in hippocampal synaptic plasticity and spatial memory processes. The antioxidant N-acetylcysteine (NAC) prevents these deleterious effects of AßOs in vitro. The main contribution of the present work is to show that AßOs injections directly into the hippocampus, by engaging oxidation-mediated reversible pathways significantly decreased RyR2 protein content but increased single RyR2 channel activation by Ca2+ and caused considerable spatial memory deficits. AßOs injections into the CA3 hippocampal region impaired rat performance in the Oasis maze spatial memory task, decreased hippocampal glutathione levels and overall content of plasticity-related proteins (c-Fos, Arc, and RyR2) and increased ERK1/2 phosphorylation. In contrast, in hippocampus-derived mitochondria-associated membranes (MAM) AßOs injections increased RyR2 levels. Rats fed with NAC for 3-weeks prior to AßOs injections displayed comparable redox potential, RyR2 and Arc protein contents, similar ERK1/2 phosphorylation and RyR2 single channel activation by Ca2+ as saline-injected (control) rats. NAC-fed rats subsequently injected with AßOs displayed the same behavior in the spatial memory task as control rats. Based on the present in vivo results, we propose that redox-sensitive neuronal RyR2 channels partake in the mechanism underlying AßOs-induced memory disruption in rodents.

High-Fat-Diet-Induced Obesity Produces Spontaneous Ventricular Arrhythmias and Increases the Activity of Ryanodine Receptors in Mice.

Ventricular arrhythmias are a common cause of sudden cardiac death, and their occurrence is higher in obese subjects. Abnormal gating of ryanodine receptors (RyR2), the calcium release channels of the sarcoplasmic reticulum, can produce ventricular arrhythmias. Since obesity promotes oxidative stress and RyR2 are redox-sensitive channels, we investigated whether the RyR2 activity was altered in obese mice. Mice fed a high fat diet (HFD) became obese after eight weeks and exhibited a significant increase in the occurrence of ventricular arrhythmias. Single RyR2 channels isolated from the hearts of obese mice were more active in planar bilayers than those isolated from the hearts of the control mice. At the molecular level, RyR2 channels from HFD-fed mice had substantially fewer free thiol residues, suggesting that redox modifications were responsible for the higher activity. Apocynin, provided in the drinking water, completely prevented the appearance of ventricular arrhythmias in HFD-fed mice, and normalized the activity and content of the free thiol residues of the protein. HFD increased the expression of NOX4, an isoform of NADPH oxidase, in the heart. Our results suggest that HFD increases the activity of RyR2 channels via a redox-dependent mechanism, favoring the appearance of ventricular arrhythmias.

Effects of breathing type on electromyographic activity of respiratory muscles at different body positions.

AIM: To compare the effect of breathing type on the activity of respiratory muscles at different body positions. METHODOLOGY: Two groups of 20 subjects each, one with upper costal and the other with costodiaphragmatic breathing, were studied. Electromyographic activity of sternocleidomastoid (SCM), diaphragm (DIA), external intercostal (EIC), and latissimus dorsi (LAT) muscles was recorded at standing and lateral decubitus positions during swallowing and maximal voluntary clenching. RESULTS: All muscles showed higher activity during standing in upper costal breathing subjects except the SCM muscle. EIC activity was higher during standing in the costodiaphragmatic breathing group. Subjects with upper costal breathing showed higher DIA activity than subjects with costodiaphragmatic breathing at both body positions and higher SCM activity at lateral decubitus position, whereas, EIC activity was only higher during swallowing. CONCLUSIONS: Subjects with upper costal breathing presented higher respiratory effort than subjects with costodiaphragmatic breathing, being most prominent at the lateral decubitus position.

Comparison of muscle activity between subjects with or without lip competence: Electromyographic activity of lips, supra- and infrahyoid muscles.

AIM: This study compares the electromyographic (EMG) activity of the muscles from the lips and hyoid bone in subjects with or without lip competence. METHODOLOGY: Two groups of 20 subjects each, with or without lip competence were studied. EMG activity of the superior orbicularis oris (SOO), inferior orbicularis oris (IOO), suprahyoid (SH) and infrahyoid (IH) muscles was recorded with the subject seated in the upright position during the following tasks: (1) at rest; (2) speaking; (3) swallowing; (4) forced deep breathing; (5) maximal voluntary clenching; and (6) chewing. RESULTS: EMG activity was significantly higher in subjects without competent lips than with competent lips in the SOO and IOO muscles during tasks 3 and 4, SOO during task 2 and IOO during task 6. EMG activity was similar in the SOO and IOO muscles during tasks 1 and 5, SOO during task 6 and IOO during task 2. Activity of the SH and IH muscles was similar in both groups for all tasks. CONCLUSIONS: Higher activity in subjects without competent lips implies a higher muscular effort due to the requirement of lip sealing during functional activities. Hyoid muscular activity was not modified by the presence or absence of lip competence.

Natural mediotrusive contact: does it affect the masticatory and neck EMG activity during tooth grinding?

OBJECTIVES: There is scarce knowledge regarding the influence of a natural mediotrusive contact on mandibular and cervical muscular activity. The purpose of this study was to analyze the EMG activity of the anterior temporalis (AT) and sternocleidomastoid (SCM) muscles during awake grinding in healthy subjects with or without a natural mediotrusive occlusal contact. METHOD: Fifteen subjects with natural mediotrusive occlusal contact (Group 1) and 15 subjects without natural mediotrusive occlusal contact (Group 2) participated. Bilateral surface EMG activity of AT and SCM muscles was recorded during unilateral eccentric or concentric tooth grinding tasks. EMG activity was normalized against the activity recorded during maximal voluntary clenching in intercuspal position (IP) for AT muscles and during maximal intentional isometric head-neck rotation to each side, for SCM muscles. RESULTS: EMG activity of AT and SCM muscles showed no statistical difference between groups. EMG activity of AT muscle was higher in the working side (WS) than in the non-WS (NWS) in Group 1 during concentric grinding (0.492 vs 0.331, p = 0.047), whereas no difference was observed in Group 2. EMG activity of SCM was similar between working and NWSs in both groups and tasks. Asymmetry indexes (AIs) were not significantly different between groups. DISCUSSION: These findings in healthy subjects support the assumption that during awake tooth grinding, central nerve control predominates over peripheral inputs, and reinforce the idea of a functional link between the motor-neuron pools that control jaw and neck muscles.

Effect of natural mediotrusive contact on electromyographic activity of jaw and cervical muscles during chewing.

OBJECTIVE: This study evaluated the effect of a natural mediotrusive contact on the electromyographic (EMG) activity of the anterior temporalis and sternocleidomastoid muscles during chewing in healthy subjects. MATERIALS AND METHODS: The study sample included two groups of 15 subjects each (Group 1: with natural mediotrusive contact; Group 2: without natural mediotrusive contact). Bilateral surface EMG activity was recorded on anterior temporalis and sternocleidomastoid muscles during unilateral chewing of a half cookie and unilateral chewing of a piece of apple. Anterior temporalis and sternocleidomastoid muscle activity was normalized against activity recorded during maximal voluntary clenching in intercuspal position and maximal intentional isometric head-neck rotation to each side, respectively. The partial and total asymmetry indexes were also calculated. Data were analyzed using Mann-Whitney, Wilcoxon and unpaired t-test. RESULTS: EMG activity of anterior temporalis and sternocleidomastoid muscles showed no significant difference between the groups. EMG activity of anterior temporalis was similar between working and non-working sides during chewing in both groups. EMG activity of sternocleidomastoid muscle was higher in the working side than in the non-working side in Group 2 subjects. Asymmetry indexes were not significantly different between groups. CONCLUSIONS: The similar EMG pattern and asymmetry indexes observed suggest the predominance of central nervous control over peripheral inputs on anterior temporalis and sternocleidomastoid motor neuron pools.

Stimulation of NOX2 in isolated hearts reversibly sensitizes RyR2 channels to activation by cytoplasmic calcium.

The response of ryanodine receptor (RyR) channels to cytoplasmic free calcium concentration ([Ca(2+)]) is redox sensitive. Here, we report the effects of a mild oxidative stress on cardiac RyR (RyR2) channels in Langendorff perfused rat hearts. Single RyR2 channels from control ventricles displayed the same three responses to Ca(2+) reported in other mammalian tissues, characterized by low, moderate, or high maximal activation. A single episode of 5 min of global ischemia, followed by 1 min of reperfusion, enhanced 2.3-fold the activity of NOX2 compared to controls and changed the frequency distribution of the different responses of RyR2 channels to calcium, favoring the more active ones: high activity response increased and low activity response decreased with respect to controls. This change was fully prevented by perfusion with apocynin or VAS 2870 before ischemia and totally reversed by the extension of the reperfusion period to 15 min. In vitro activation of NOX2 in control SR vesicles mimicked the effect of the ischemia/reperfusion episode on the frequencies of emergence of single RyR2 channel responses to [Ca(2+)] and increased 2.2-fold the rate of calcium release in Ca(2+)-loaded SR vesicles. In vitro changes were reversed at the single channel level by DTT and in isolated SR vesicles by glutaredoxin. Our results indicate that in whole hearts a mild oxidative stress enhances the response of cardiac RyR2 channels to calcium via NOX2 activation, probably by S-glutathionylation of RyR2 protein. This change is transitory and fully reversible, suggesting a possible role of redox modification in the physiological response of cardiac RyR2 to cellular calcium influx.

Redox-sensitive stimulation of type-1 ryanodine receptors by the scorpion toxin maurocalcine.

The scorpion toxin maurocalcine acts as a high affinity agonist of the type-1 ryanodine receptor expressed in skeletal muscle. Here, we investigated the effects of the reducing agent dithiothreitol or the oxidizing reagent thimerosal on type-1 ryanodine receptor stimulation by maurocalcine. Maurocalcine addition to sarcoplasmic reticulum vesicles actively loaded with calcium elicited Ca²âº release from native vesicles and from vesicles pre-incubated with dithiothreitol; thimerosal addition to native vesicles after Ca²âº uptake completion prevented this response. Maurocalcine enhanced equilibrium [³H]-ryanodine binding to native and to dithiothreitol-treated reticulum vesicles, and increased 5-fold the apparent Ki for Mg²âº inhibition of [³H]-ryanodine binding to native vesicles. Single calcium release channels incorporated in planar lipid bilayers displayed a long-lived open sub-conductance state after maurocalcine addition. The fractional time spent in this sub-conductance state decreased when lowering cytoplasmic [Ca²âº] from 10 µM to 0.1 µM or at cytoplasmic [Mg²âº]≥30 µM. At 0.1 µM [Ca²âº], only channels that displayed poor activation by Ca²âº were readily activated by 5 nM maurocalcine; subsequent incubation with thimerosal abolished the sub-conductance state induced by maurocalcine. We interpret these results as an indication that maurocalcine acts as a more effective type-1 ryanodine receptor channel agonist under reducing conditions.

Age-dependent increases in apoptosis/necrosis ratios in human lymphocytes exposed to oxidative stress.

Unlike apoptosis, mechanisms leading to necrosis are less well understood. Moreover, changes in necrosis as a function of age have not been studied in human lymphocytes. H(2)O(2)-induced death of peripheral lymphocytes (56 healthy donors, 24-95 years) was evaluated by flow cytometry and propidium iodide staining, caspase activation, DNA laddering, and electron microscopy. H(2)O(2)-induced stress was associated with high levels of necrosis in young individuals (≤30 years), whereas progressively enhanced apoptotic death was observed in older donors, without changes in overall lymphocyte survival. Thus, apoptosis/necrosis ratios were inverted in young versus elderly (≥65 years) donors. Death was not accompanied by increased caspase activity and, accordingly, unaffected by caspase inhibition; however, it was almost completely prevented by poly ADP ribose polymerase inhibition. In summary, aging was associated with changes in the apoptosis/necrosis ratios, rather than susceptibility per se to H(2)O(2)-induced death, which was caspase independent but poly ADP ribose polymerase dependent. Understanding this switch in death modes may aid in understanding age-related disorders.

Modulation of cardiac ryanodine receptor activity by ROS and RNS.

Calcium release through cardiac ryanodine receptors (RyR2) triggers heart muscle contraction. Reactive oxygen/nitrogen species (ROS/RNS), normally produced in the heart, promote endogenous RyR2 S-nitrosylation and S-glutathionylation. These reversible redox modifications increase RyR2 activity in vitro, and presumably also in vivo. RyR2 S-glutathionylation increases under physiologically relevant conditions (tachycardia and exercise), suggesting that cardiac cells utilize this redox modification to increase RyR2 activity under increased demand. In contrast, in vivo changes in RyR2 S-nitrosylation in response to physiological stimuli remain uncharacterized. The number and identity of the highly reactive RyR2 cysteine residues and the nature of the redox modification they undergo are presently unknown. Likewise, the physiological sources of ROS/RNS responsible for functionally relevant RyR2 redox modifications have not been completely identified. The redox state of RyR2 is altered in heart failure leading to enhanced RyR2 activity, which presumably contributes to decrease SR calcium content and induce other calcium release abnormalities observed in heart failure. Greater understanding of RyR2 redox modulation is necessary to counteract the deleterious consequences of RyR2 activity deregulation caused by oxidative stress.

Ischemia enhances activation by Ca2+ and redox modification of ryanodine receptor channels from rat brain cortex.

Cerebral ischemia stimulates Ca2+ influx and thus increases neuronal intracellular free [Ca2+]. Using a rat model of cerebral ischemia without recirculation, we tested whether ischemia enhances the activation by Ca2+ of ryanodine receptor (RyR) channels, a requisite feature of RyR-mediated Ca2+-induced Ca2+ release (CICR). To this aim, we evaluated how single RyR channels from endoplasmic reticulum vesicles, fused into planar lipid bilayers, responded to cytoplasmic [Ca2+] changes. Endoplasmic reticulum vesicles were isolated from the cortex of rat brains incubated without blood flow for 5 min at 37 degrees C (ischemic) or at 4 degrees C (control). Ischemic brains displayed increased oxidative intracellular conditions, as evidenced by a lower ratio (approximately 130:1) of reduced/oxidized glutathione than controls (approximately 200:1). Single RyR channels from ischemic or control brains displayed the same three responses to Ca2+ reported previously, characterized by low, moderate, or high maximal activity. Relative to controls, RyR channels from ischemic brains displayed with increased frequency the high activity response and with lower frequency the low activity response. Both control and ischemic cortical vesicles contained the RyR2 and RyR3 isoforms in a 3:1 proportion, with undetectable amounts of RyR1. Ischemia reduced [3H]ryanodine binding and total RyR protein content by 35%, and increased at least twofold endogenous RyR2 S-nitrosylation and S-glutathionylation without affecting the corresponding RyR3 endogenous levels. In vitro RyR S-glutathionylation but not S-nitrosylation favored the emergence of high activity channels. We propose that ischemia, by enhancing RyR2 S-glutathionylation, allows RyR2 to sustain CICR; the resulting amplification of Ca2+ entry signals may contribute to cortical neuronal death.

Effects of ATP, Mg2+, and redox agents on the Ca2+ dependence of RyR channels from rat brain cortex.

Despite their relevance for neuronal Ca(2+)-induced Ca(2+) release (CICR), activation by Ca(2+) of ryanodine receptor (RyR) channels of brain endoplasmic reticulum at the [ATP], [Mg(2+)], and redox conditions present in neurons has not been reported. Here, we studied the effects of varying cis-(cytoplasmic) free ATP concentration ([ATP]), [Mg(2+)], and RyR redox state on the Ca(2+) dependence of endoplasmic reticulum RyR channels from rat brain cortex. At pCa 4.9 and 0.5 mM adenylylimidodiphosphate (AMP-PNP), increasing free [Mg(2+)] up to 1 mM inhibited vesicular [(3)H]ryanodine binding; incubation with thimerosal or dithiothreitol decreased or enhanced Mg(2+) inhibition, respectively. Single RyR channels incorporated into lipid bilayers displayed three different Ca(2+) dependencies, defined by low, moderate, or high maximal fractional open time (P(o)), that depend on RyR redox state, as we have previously reported. In all cases, cis-ATP addition (3 mM) decreased threshold [Ca(2+)] for activation, increased maximal P(o), and shifted channel inhibition to higher [Ca(2+)]. Conversely, at pCa 4.5 and 3 mM ATP, increasing cis-[Mg(2+)] up to 1 mM inhibited low activity channels more than moderate activity channels but barely modified high activity channels. Addition of 0.5 mM free [ATP] plus 0.8 mM free [Mg(2+)] induced a right shift in Ca(2+) dependence for all channels so that [Ca(2+)] <30 microM activated only high activity channels. These results strongly suggest that channel redox state determines RyR activation by Ca(2+) at physiological [ATP] and [Mg(2+)]. If RyR behave similarly in living neurons, cellular redox state should affect RyR-mediated CICR.

Redox regulation of RyR-mediated Ca2+ release in muscle and neurons.

Changes in the redox state of the intracellular ryanodine receptor/Ca2+ release channels of skeletal and cardiac muscle or brain cortex neurons affect their activity. In particular, agents that oxidize or alkylate free SH residues of the channel protein strongly enhance Ca(2+)-induced Ca2+ release, whereas reducing agents have the opposite effects. We will discuss here how modifications of highly reactive cysteine residues by endogenous redox agents or cellular redox state influence RyR channel activation by Ca2+ and ATP or inhibition by Mg2+. Possible physiological and pathological implications of these results on cellular Ca2+ signaling will be addressed as well.

Redox regulation of RyR-mediated Ca2+ release in muscle and neurons

Changes in the redox state of the intracellular ryanodine receptor/Ca2+ release channels of skeletal and cardiac muscle or brain cortex neurons affect their activity. In particular, agents that oxidize or alkylate free SH residues of the channel protein strongly enhance Ca2+-induced Ca2+ release, whereas reducing agents have the opposite effects. We will discuss here how modifications of highly reactive cysteine residues by endogenous redox agents or cellular redox state influence RyR channel activation by Ca2+ and ATP or inhibition by Mg2+. Possible physiological and pathological implications of these results on cellular Ca2+ signaling will be addressed as well.

SH oxidation coordinates subunits of rat brain ryanodine receptor channels activated by calcium and ATP.

We have reported that ryanodine receptor (RyR) channels display three different responses to cytoplasmic free Ca2+ concentration ([Ca2+]) depending on their redox state (Marengo JJ, Hidalgo C, and Bull R. Biophys J 74: 1263-1277, 1998), with low, moderate, and high maximal fractional open times (Po). Activation by ATP of single RyR channels from rat brain cortex was tested in planar lipid bilayers with 10 or 0.1 microM cytoplasmic [Ca2+]. At 10 microM [Ca2+], low-Po channels presented lower apparent affinity to activation by ATP [[ATP] for half-maximal activation (KaATP) = 422 microM] than moderate-Po channels (KaATP = 82 microM). Oxidation of low-Po channels with thimerosal or 2,2'-dithiodipyridine (DTDP) gave rise to moderate-Po channels and decreased KaATP from 422 to 82 microM. At 0.1 microM cytoplasmic [Ca2+], ATP induced an almost negligible activation of low-Po channels. After oxidation to high-Po behavior, activation by ATP was markedly increased. Noise analysis of single-channel fluctuations of low-Po channels at 10 microM [Ca2+] plus ATP revealed the presence of subconductance states, suggesting a conduction mechanism that involves four independent subchannels. On oxidation the subchannels opened and closed in a concerted mode.