Intrathecal application of short interfering RNA knocks down c-jun expression and augments spinal motoneuron death after root avulsion in adult rats.

The immediate-early gene, c-jun, is expressed in spinal motoneurons after spinal root avulsion. The expression of c-jun was suggested to be necessary for motoneuron survival and regeneration after avulsion. In the present study, a small interfering RNA (siRNA) was delivered intrathecally to the injured spinal segments immediately after root avulsion in rats to knock down expression of the c-jun gene in injured spinal motoneurons so as to explore the role of c-jun in the motoneurons in vivo. Our results showed that the siRNA not only inhibited the expression of both c-jun mRNA and protein but also augmented the death of injured motoneurons at day 14 post-injury. These findings indicated that induction of c-jun gene expression plays a pivotal role in the survival of injured motoneurons. Meanwhile, these results suggest that siRNAs applied intrathecally can effectively mediate the expression of the c-jun gene in injured motoneurons.

Unilateral low-frequency stimulation of central piriform cortex delays seizure development induced by amygdaloid kindling in rats.

Low-frequency stimulation of the kindling site interferes with the course of kindling epileptogenesis. The present study examined the effect of unilateral low-frequency stimulation of the central piriform cortex on seizure development induced by amygdaloid kindling in rats. The ipsilateral or contralateral central piriform cortex received low-frequency stimulation (15 min train of 0.1 ms pulses at 1 Hz and 50-150 muA) immediately after termination of once daily kindling stimulation (2 s train of 1 ms pulses at 60 Hz and 150-300 microA) in the right amygdala for 30 days. Low-frequency stimulation of either the ipsilateral or contralateral central piriform cortex significantly suppressed the progression of seizure stages and reduced afterdischarge duration throughout the course of amygdaloid kindling. The marked suppression induced by low-frequency stimulation of the central piriform cortex on either side was predominantly due to the significant retardation of progression from stage 0 to stage 1 and stage 3 to stage 4 seizures. In addition, the suppressive effect of low-frequency stimulation did not disappear when the stimulation was stopped; it could persist for at least 10 days. These findings indicate that brain areas other than the kindling focus, such as the central piriform cortex on both sides, can also be used as reasonable targets for low-frequency stimulation to retard seizure development induced by amygdaloid kindling. Secondly, like the ipsilateral central piriform cortex, the contralateral central piriform cortex may also participate in the progression and secondary generalization of focal seizures. The study suggests that unilateral low-frequency stimulation of the central piriform cortex may have a significant antiepileptogenic effect, and may be helpful for exploring effective and long-lasting therapies for human temporal lobe epilepsy.

Luteolin induces vasorelaxion in rat thoracic aorta via calcium and potassium channels.

The aims of the present study were to investigate the vasoactive effects of luteolin and its mechanisms of action on the rat thoracic aorta. Luteolin (4.5-36 micromol/L) caused a concentration-dependent relaxation of endothelium-intact or endothelium-denuded aortic rings precontracted with phenylephrine (PE, 10(-6) mol/L) or a high level of K+ (6 x 10(-2) mol/L). Luteolin induced a shift of the PE concentration-response curve to the right and downward. L-NAME and propranolol did not influence the vascular effect of luteolin. However, 5-hydroxydecanoate, tetraethylammonium, BaCl2 and 4-aminopyridine significantly attenuated the vasorelaxant effect of luteolin. In Ca2+ -free medium, medium with graded concentrations of Ca2+, or K+ -free solution, luteolin reduced PE-induced contraction. It is concluded that luteolin induces endothelium-independent relaxation in rat thoracic aorta. The mechanism involves the inhibition of sarcolemmal Ca2+ channels, release from intracellular Ca2+ stores and activation of K+ channels.

Dual inhibition of 5-LOX and COX-2 suppresses colon cancer formation promoted by cigarette smoke.

Previous studies indicate that the arachidonic acid-metabolizing enzymes COX-2 and 5-LOX are overexpressed during the process of colonic adenoma formation promoted by cigarette smoke. The aims of the present study were to investigate whether there exists a relationship between COX-2 and 5-LOX, and whether dual inhibition of COX-2 and 5-LOX has an anticarcinogenic effect in the colonic tumorigenesis promoted by cigarette smoke. Results showed that pretreating colon cancer cells with cigarette smoke extract (CSE) promoted colon cancer growth in the nude mouse xenograft model. Inhibition of COX-2 or 5-LOX reduced the tumor size. In the group treated with COX-2-inhibitor, the PGE2 level decreased while the LTB4 level increased. In contrast, in the 5-LOX-inhibitor treated group, the LTB4 level was reduced and the PGE2 level was unchanged. However, combined treatment with both COX-2 and 5-LOX inhibitors further inhibited the tumor growth promoted by CSE over treatment with either COX-2-inhibitor or 5-LOX-inhibitor alone. This was accompanied by the downregulation of PGE2 and LTB4. In an in vitro study, we found that the action of CSE on colon cancer cells was mediated by 5-LOX DNA demethylation. In summary, these results indicate that inhibition of COX-2 may lead to a shunt of arachidonic acid metabolism towards the leukotriene pathway during colonic tumorigenesis promoted by CSE. Suppression of 5-LOX did not induce such a shunt and produced a better response. Therefore, 5-LOX inhibitor is more effective than COX-2 inhibitor, and blocker of both COX-2 and 5-LOX may present a superior anticancer profile in cigarette smokers.

Early association of electrocardiogram alteration with infarct size and cardiac function after myocardial infarction.

OBJECTIVE: Myocardial infarction (MI) is the main cause of heart failure, but the relationship between the extent of MI and cardiac function has not been clearly determined. The present study was undertaken to investigate early changes in the electrocardiogram associated with infarct size and cardiac function after MI. METHODS: MI was induced by ligating the left anterior descending coronary artery in rats. Electrocardiograms, echocardiographs and hemodynamic parameters were assessed and myocardial infarct size was measured from mid-transverse sections stained with Masson's trichrome. RESULTS: The sum of pathological Q wave amplitudes was strongly correlated with myocardial infarct size (r = 0.920, P < 0.0001), left ventricular ejection fraction (r = -0.868, P < 0.0001) and left ventricular end diastolic pressure (r = 0.835, P < 0.0004). Furthermore, there was close relationship between MI size and cardiac function as assessed by left ventricular ejection fraction (r = -0.913, P < 0.0001) and left ventricular end diastolic pressure (r = 0.893, P < 0.0001). CONCLUSION: The sum of pathological Q wave amplitudes after MI can be used to estimate the extent of MI as well as cardiac function.

The mitochondrial calcium uniporter participates in ischemia/reperfusion injury and in cardioprotection by ischemic preconditioning.

The objective of the present study was to determine whether the mitochondrial calcium uniporter plays a role in cardioprotection by ischemic preconditioning (IPC). Isolated rat hearts were subjected to 30 min regional ischemia by ligation of the left anterior descending artery followed by 120 min reperfusion. We found that both IPC and inhibition of the mitochondrial calcium uniporter during reperfusion improved recovery of left ventricular developed pressure, maximal rise velocity and end-diastolic pressure, and reduced infarct size and lactate dehydrogenase release. These protective effects were attenuated by activating the mitochondrial calcium uniporter. We conclude that the mitochondrial calcium uniporter is involved in the cardioprotection of ischemic preconditioning.

Role of the mitochondrial permeability transition pore in TNF-alpha-induced recovery of ventricular contraction and reduction of infarct size in isolated rat hearts subjected to ischemia/reperfusion.

Pretreatment with tumor necrosis factor-alpha (TNF-alpha) is known to trigger cardioprotection. TNF-alpha can activate multiple downstream signaling cascades. However, it is not known whether the mitochondrial permeability transition pore (MitoPTP) is involved in TNF-alpha-induced cardioprotection. In the present study, we examined whether TNF-alpha inhibits MitoPTP opening. In isolated rat hearts subjected to 30 min regional ischemia and 120 min reperfusion, pretreatment with 10 U/ml TNF-alpha for 7 min followed by 10 min washout improved the recovery of left ventricular developed pressure (LVDP) and rate-pressure product (RPP = LVDP x heart rate) during reperfusion and reduced the infarct size. Administration of 20 micromol/L atractyloside, a MitoPTP opener, for 20 min (last 5 min of ischemia and first 15 min of reperfusion) and pretreatment with 1 mu inhibitor of the Ca2+-activated K+ mol/L paxilline, an channel, for 5 min before ischemia, attenuated the recovery of LVDP and RPP and the reduction of infarct size induced by TNF-alpha. The findings indicate that, in the isolated heart model, TNF-alpha protects myocardium against ischemia/reperfusion injury via inhibiting MitoPTP opening as well as by activating the Ca2+-activated K+ channel.

The use of artificial neural networks in the motor program.

Though it is commonly assumed that the brain creates "motor programs" which store the information essential to perform a motor skill, little direct evidence exists for such motor programs. Electromyography (EMG) provides a look into the motoneurons--level of a movement by measuring the electrical activity in relation to the muscle's involvement in the movement In this paper, artificial neural networks (ANNs) were applied to define the temporal patterns of EMG activity used by normal subjects in performing step-tracking tasks, and how such patterns change with practice. Our results demonstrate that ANNs could be trained to detect the input-output relationship between muscles' onset times and reaction times, and provided evidence to support the existence of a motor program.

Dorsal column is not involved in the mechanism of the hypotensive effect by simulating acupuncture on rat hindlimb.

The present study investigated the role of the dorsal column (DC) in the mechanism of the hypotensive effect induced by simulating acupuncture on rat hindlimb. The femoral arterial pressure and electrocardiogram (ECG) of rats were recorded when the hypothalamic paraventricular nucleus (PVN) was electrically stimulated with or without DC lesion. Stimulation of the deep peroneal nerve (DPN) decreased the pressor response elicited by electrical stimulation of the PVN. Thirty minutes after micro-dissection of the right DC, the inhibitory effect of stimulating the right or left DPN on the pressor response induced by stimulation of the contralateral PVN was not altered. Of 6 rats tested, the inhibitory effect of stimulating the right or left DPN could still be observed five days after the right DC was destroyed. The pain responses of both hindlimbs of the rats with the right DC destroyed showed no obvious difference when compared with the sham control rats. These data suggest that the DC is not involved in the inhibitory effect of stimulating the DPN on the pressor response induced by the PVN activation.

Influence of interleukin-2 on Ca2+ handling in rat ventricular myocytes.

In the present study, we examined the effect of interleukin-2 (IL-2) on cardiomyocyte Ca(2+) handling. The effects of steady-state and transient changes in stimulation frequency on the intracellular Ca(2+) transient were investigated in isolated ventricular myocytes by spectrofluorometry. In the steady state (0.2 Hz) IL-2 (200 U/ml) decreased the amplitude of Ca(2+) transients induced by electrical stimulation and caffeine. At 1.25 mM extracellular Ca(2+) concentration ([Ca(2+)](o)), when the stimulation frequency increased from 0.2 to 1.0 Hz, diastolic Ca(2+) level and peak intracellular Ca(2+) concentration ([Ca(2+)](i)), as well as the amplitude of the transient, increased. The positive frequency relationships of the peak and amplitude of [Ca(2+)](i) transients were blunted in the IL-2-treated myocytes. The effect of IL-2 on the electrically induced [Ca(2+)](i) transient was not normalized by increasing [Ca(2+)](o) to 2.5 mM. IL-2 inhibited the frequency relationship of caffeine-induced Ca(2+) release. Blockade of sarcoplasmic reticulum (SR) Ca(2+)-ATPase with thapsigargin resulted in a significant reduction of the amplitude-frequency relationship of the transient similar to that induced by IL-2. The restitutions were not different between control and IL-2 groups at 1.25 mM [Ca(2+)](o), which was slowed in IL-2-treated myocytes when [Ca(2+)](o) was increased to 2.5 mM. There was no difference in the recirculation fraction (RF) between control and IL-2-treated myocytes at both 1.25 and 2.5 mM [Ca(2+)](o). The effects of IL-2 on frequency relationship, restitution, and RF may be due to depressed SR functions and an increased Na(+)-Ca(2+) exchange activity, but not to any change in L-type Ca(2+) channels.

A phenomenological model for the responses of auditory-nerve fibers: I. Nonlinear tuning with compression and suppression.

A phenomenological model was developed to describe responses of high-spontaneous-rate auditory-nerve (AN) fibers, including several nonlinear response properties. Level-dependent gain (compression), bandwidth, and phase properties were implemented with a control path that varied the gain and bandwidth of tuning in the signal-path filter. By making the bandwidth of the control path broad with respect to the signal path, the wide frequency range of two-tone suppression was included. By making the control-path filter level dependent and tuned to a frequency slightly higher than the signal-path filter, other properties of two-tone suppression were also included. These properties included the asymmetrical growth of suppression above and below the characteristic frequency and the frequency offset of the suppression tuning curve with respect to the excitatory tuning curve. The implementation of this model represents a relatively simple phenomenological description of a single mechanism that underlies several important nonlinear response properties of AN fibers. The model provides a tool for studying the roles of these nonlinearities in the encoding of simple and complex sounds in the responses of populations of AN fibers.

Renewal-process approximation of a stochastic threshold model for electrical neural stimulation.

In a recent set of modeling studies we have developed a stochastic threshold model of auditory nerve response to single biphasic electrical pulses (Bruce et al., 1999c) and moderate rate (less than 800 pulses per second) pulse trains (Bruce et al., 1999a). In this article we derive an analytical approximation for the single-pulse model, which is then extended to describe the pulse-train model in the case of evenly timed, uniform pulses. This renewal-process description provides an accurate and computationally efficient model of electrical stimulation of single auditory nerve fibers by a cochlear implant that may be extended to other forms of electrical neural stimulation.

The effects of food fragmentation index on mandibular closing angle in human mastication.

Jaw movements were recorded using a three-dimensional magnetic sensing system (Sirognathograph, Siemens) in 10 human volunteers while chewing standardized volumes of 15 food types, ranging from soft cheeses to hard nuts. The maximum horizontal amplitude inside the chewing loop (width of the loop), the height of the loop (vertical amplitude) and the closing angle made by the jaw during the late closing phase of the cycle relative to the vertical were calculated. The most highly significant correlation was between the closing angle and the square root of the ratio of two mechanical properties of the food - the toughness (R) and modulus of elasticity (E)(r = -0.85, p<0.001). The width of the chewing loop was also significantly correlated with the above combination of food properties (r = -0.75, p<0.01) whereas the vertical amplitude was not. Thus, the mechanical properties of foods appear to influence the pattern of mandibular movements in human mastication similarly to their reported effect on food breakdown rates and anterior temporalis activity during mastication.

The effects of stochastic neural activity in a model predicting intensity perception with cochlear implants: low-rate stimulation.

Most models of auditory nerve response to electrical stimulation are deterministic, despite significant physiological evidence for stochastic activity. Furthermore, psychophysical models and analyses of physiological data using deterministic descriptions do not accurately predict many psychophysical phenomena. In this paper we investigate whether inclusion of stochastic activity in neural models improves such predictions. To avoid the complication of interpulse interactions and to enable the use of a simpler and faster auditory nerve model we restrict our investigation to single pulses and low-rate (< 200 pulses/s) pulse trains. We apply signal detection theory to produce direct predictions of behavioral threshold, dynamic range and intensity difference limen. Specifically, we investigate threshold versus pulse duration (the strength-duration characteristics), threshold and uncomfortable loudness (and the corresponding dynamic range) versus phase duration, the effects of electrode configuration on dynamic range and on strength-duration, threshold versus number of pulses (the temporal-integration characteristics), intensity difference limen as a function of loudness, and the effects of neural survival on these measures. For all psychophysical measures investigated, the inclusion of stochastic activity in the auditory nerve model was found to produce more accurate predictions.

A stochastic model of the electrically stimulated auditory nerve: single-pulse response.

Most models of neural response to electrical stimulation, such as the Hodgkin-Huxley equations, are deterministic, despite significant physiological evidence for the existence of stochastic activity. For instance, the range of discharge probabilities measured in response to single electrical pulses cannot be explained at all by deterministic models. Furthermore, there is growing evidence that the stochastic component of auditory nerve response to electrical stimulation may be fundamental to functionally significant physiological and psychophysical phenomena. In this paper we present a simple and computationally efficient stochastic model of single-fiber response to single biphasic electrical pulses, based on a deterministic threshold model of action potential generation. Comparisons with physiological data from cat auditory nerve fibers are made, and it is shown that the stochastic model predicts discharge probabilities measured in response to single biphasic pulses more accurately than does the equivalent deterministic model. In addition, physiological data show an increase in stochastic activity with increasing pulse width of anodic/cathodic biphasic pulses, a phenomenon not present for monophasic stimuli. These and other data from the auditory nerve are then used to develop a population model of the total auditory nerve, where each fiber is described by the single-fiber model.

A stochastic model of the electrically stimulated auditory nerve: pulse-train response.

The single-pulse model of the companion paper [1] is extended to describe responses to pulse trains by introducing a phenomenological refractory mechanism. Comparisons with physiological data from cat auditory nerve fibers are made for pulse rates between 100 and 800 pulses/s. First, it is shown that both the shape and slope of mean discharge rate curves are better predicted by the stochastic model than by the deterministic model. Second, while interpulse effects such as refractory effects do indeed increase the dynamic range at higher pulse rates, both the physiological data and the model indicate that much of the dynamic range for pulse-train stimuli is due to stochastic activity. Third, it is shown that the stochastic model is able to predict the general magnitude and behavior of variance in discharge rate as a function of pulse rate, while the deterministic model predicts no variance at all.

Food properties that influence neuromuscular activity during human mastication.

The rate of breakdown of food in mastication depends on the ratio of two mechanical properties of the food--the toughness and modulus of elasticity (Agrawal et al., 1997)--a result which can be predicted by an analysis of the energetics of fracture. The work input to produce food fragmentation is provided by the masticatory muscles, the activity levels of which depend on sensory feedback from the mouth. Here, we test the hypothesis that the activity of a representative of this musculature is modulated by the above combination of food properties. The surface electrical activity (EMG) of the anterior temporalis muscles of ten human subjects was recorded while subjects chewed standardized volumes of 15 food types. The integrated EMG in these muscles was highly significantly related to the square root of the ratio of the above two food properties. Significant correlations were found between this food property index and integrated EMG, both when data for all chews and all subjects were lumped together (r = -0.86; p < 0.0001) and when correlation coefficients between the index and EMG were plotted for each chew made by each subject. Except for two subjects in the first chew, these coefficients reached and maintained highly significant levels throughout the masticatory sequence. Thus, a clear relationship between the electrical activity of a jaw-closing muscle and the mechanical properties of food has been found for the first time.

Electromyographic activity in a distant muscle during simple voluntary movements: an unexpected hand-eye linkage.

PURPOSE: Supraspinal sensorimotor maps contain adjacent representations of the hand and face. These experiments were designed to determine whether interactions between the representations could be detected at the periphery. METHODS: Simultaneous EMG recordings were made from orbicularis oculi and wrist flexor during three conditions, with 50 repetitions each: during exposure to an auditory cue; responding to the auditory cue by wrist flexion; and performing a self-paced wrist flexion in the absence of an auditory cue. RESULTS: In 8 of 10 healthy volunteers, the rectified, averaged EMGs showed small but consistent increases in orbicularis oculi activity during wrist flexion in the two tasks involving that movement, but not in the auditory task alone. CONCLUSION: In one or more of the central sensorimotor maps, voluntary activation of the wrist flexor representation is accompanied by liminal activation of a nearby facial representation which projects to the motoneurons of orbicularis oculi.

Mechanical properties of foods responsible for resisting food breakdown in the human mouth.

The fragmentation of foods (breakage function) was measured in five humans on "bagged' single particles of 28 foods from three food groups. The change in the square root of the specific surface of the particles (the specific surface being the area of particle silhouettes, measured by image analysis, divided by original particle volume) produced by one bite, averaged for all participants, was inversely linearly related to the square root of the toughness of the foods divided by the square root of their Young's moduli(r = -0.86; p < 0.00001). This relation is predicted by an analysis based on food fragmentation within a limited jaw displacement. Thus, resistance to jaw movement appears to provide sensory information on the deformation fracture and fragmentation of foods. It is believed that this is the first time that a relation between the breakage of food particles by the teeth and their material properties has been found, and the finding has considerable implications for human masticatory studies, for the analysis of dentition and diet in mammals and for texture studies in food science.