Regulation of intracellular Ca2+ oscillation in AR42J cells.

Recordings of [Ca2+]i in single AR42J cells loaded with Fura 2 were used to study regulation of [Ca2+]i oscillation. Continuous stimulation with the cholecystokinin analogue, (t-butyloxycarbonyl-Tyr-(SO3)-norleucine-Gly-Trp-Nle-Asp-2-phenylethyl ester) or carbachol evoked long lasting oscillation in [Ca2+]i. Removal of CCK-JMV-180 after brief stimulation did not abruptly stop the oscillation. Rather, removal of CCK-JMV-180 resulted in time-dependent reduction in amplitude with little change in frequency of oscillation. The patterns of [Ca2+]i oscillation were affected by activation of protein kinase C and protein kinase A. However, down-regulation of protein kinase C activity did not prevent stimulation of [Ca2+]i oscillation. Hence, we conclude that an active protein kinase C pathway is not crucial for [Ca2+]i oscillation in this cell line. Variation in extracellular Ca2+ concentration (Ca2+out) was used to further characterize the oscillation. Reducing Ca2+out to approximately 10 microM resulted in a time dependent inhibition of [Ca2+]i oscillation. Subsequent step increases in Ca2+out up to 2-3 mM resulted in increased amplitude and frequency of oscillation. Further increase in Ca2+out or an increase in plasma membrane permeability to Ca2+, brought about by an increase in pHo, resulted in increased amplitude, decreased frequency, and modified shape of the [Ca2+]i spikes. These observations point to the existence of regulatory mechanisms controlling the duration of Ca2+ release and entry during [Ca2+]i oscillation.     

Acoustic signals from frog skeletal muscle.

Acoustic, force, and compound muscle action-potential signals were recorded simultaneously during maximal isometric twitches of frog gastrocnemius muscles. The onset of sound production occurred after the onset of muscle depolarization but before the onset of external force production. Acoustic waveforms consisted of oscillations that initially increased in amplitude, followed by decaying oscillations. The peak-to-peak acoustic amplitude increased with increasing temperature with a Q10 of 2.6 +/- 0.2 over a range of 7.0-25.0 degrees C. The acoustic amplitude increased with increasing muscle length up to approximately 90% of the optimal length for force generation. As length was increased further, the acoustic amplitude decreased. Microphones positioned on opposite sides of the muscle recorded acoustic signals that were 180 degrees out of phase. These results provided evidence that sound production is produced by lateral oscillations of muscle. The oscillation frequency may provide a measure of mechanical properties of muscle.     

GABAergic effects on the slow oscillatory neural activities in the procerebrum of Limax valentianus. Short communication

We examined GABAergic modulation on "slow" oscillation (<1.0 Hz) of the procerebrum in the terrestrial mollusk, Limax valentianus. Short application of GABA-receptor agonists slightly increased the frequency of a periodic oscillation in the procerebrum, whereas persistent application decreased it. GABA-receptor antagonists decreased the oscillatory frequency. The GABA-like immunoreactivities were found in the neuropil and the cell body layers of the procerebrum. Because GABAergic inhibition is known to be essential for the generation of "fast" synchronous neuronal oscillation in the CNSs in othre many animals, our present findings are first evidence suggesting that GABA modulates 'slow' oscillation in the CNS.     

Physicochemical effects enhance surfactant transport in pulsatile motion of a semi-infinite bubble

In this study, we investigate the sorption of pulmonary surfactant (Infasurf, Ony, Buffalo, NY) occurring at the air-liquid interface of a semi-infinite finger of air as it oscillates and progresses along a small rigid tube () occluded with a surfactant-doped solution of concentrations . This simple experimental model of pulmonary airway reopening is designed to examine how altering the fluid flow field may lower reopening pressures and lead to a reduction in airway wall damage that is associated with the mechanical ventilation of an obstructed pulmonary system in airways of the deep lung with depleted endogenous and little exogenous surfactant. We analyzed a range of pulsatile flow scenarios by varying the oscillation frequency (), the oscillation flow waveform, and the steady flow rate (). These experimental studies indicate that a high frequency (1 Hz, amplitude = 5 mm), fast-forward oscillation waveform superimposed onto a fast steady flow () substantially reduces mean reopening pressures (31%) as a consequence of the modified flow field and the commensurate increase in surfactant transport and adsorption. This result suggests that imposing high frequency, low amplitude oscillations during airway reopening will help to diminish ventilator-induced lung injury.     

In Vitro Dose-Dependent Inhibition of the Intracellular Spontaneous Calcium Oscillations in Developing Hippocampal Neurons by Ketamine

Spatial and temporal abnormalities in the frequency and amplitude of the cytosolic calcium oscillations can impact the normal physiological functions of neuronal cells. Recent studies have shown that ketamine can affect the growth and development and even induce the apoptotic death of neurons. This study used isolated developing hippocampal neurons as its study subjects to observe the effect of ketamine on the intracellular calcium oscillations in developing hippocampal neurons and to further explore its underlying mechanism using Fluo-4-loaded laser scanning confocal microscopy. Using a semi-quantitative method to analyze the spontaneous calcium oscillatory activities, a typical type of calcium oscillation was observed in developing hippocampal neurons. In addition, the administration of NMDA (N-Methyl-D-aspartate) at a concentration of 100 µM increased the calcium oscillation amplitude. The administration of MK801 at a concentration of 40 µM inhibited the amplitude and frequency of the calcium oscillations. Our results demonstrated that an increase in the ketamine concentration, starting from 30 µM, gradually decreased the neuronal calcium oscillation amplitude. The inhibition of the calcium oscillation frequency by 300 µM ketamine was statistically significant, and the neuronal calcium oscillations were completely eliminated with the administration of 3,000 µM Ketamine. The administration of 100, 300, and 1,000 µM NMDA to the 1 mM ketamine-pretreated hippocampal neurons restored the frequency and amplitude of the calcium oscillations in a dose-dependent manner. In fact, a concentration of 1,000 µM NMDA completely reversed the decrease in the calcium oscillation frequency and amplitude that was induced by 1 mM ketamine. This study revealed that ketamine can inhibit the frequency and amplitude of the calcium oscillations in developing hippocampal neurons though the NMDAR (NMDA receptor) in a dose-dependent manner, which might highlight a possible underlying mechanism of ketamine toxicity on the rat hippocampal neurons during development.     

Temporal tuning and nonlinearity of intraretinal pathways in turtle: Effects of temperature,stimulus intensity,and size

Flash responses, amplitude and phase transfer functions, and nonlinearities were measured in turtle retina for pathways with photoreceptor inputs and outputs from horizontal (HC), hyperpolarizing bipolar (HBC), sustained amacrine (AC), and on-off ganglion (GC) cells. Flash responses slowed and attenuated in all cells as temperature decreased. Whitenoise transfer properties of sustained-type cells (HC, HBC, AC) were of low- or bandpass type; highfrequency cut-off (f _c) and phase crossover frequency decreased with temperature. f _c increased as spot diameter was increased. Nonlinearity of these sustained-response pathways (distortion product frequencies in response to a sum-of-sinusoids input probe) increases with intensity and may depend on amplitude saturation limiting. On/off GC synaptic and spike activity increased as spot diameter decreased and intensity increased. Amplitude transfer functions had a low-frequency peak (PSP activity) and monotonically decreasing amplitude vs. frequency shape (spikes and transient PSP activity). Nonlinearity increased with stimulus intensity; it was maximal with 1 mm spot size, less with smaller (500 m) and larger (5 mm) spots. It may depend on the functional equivalent of full-wave rectification (on-off response).This work was supported by NEI grant R01 EY03383     

固态发酵的参数周期变化及对微生物发酵的影响

研究了压力脉动固态发酵反应器内环境参数的周期性变化以及这些周期性的环境刺激对于固态培养的斜卧青霉发酵的影响。研究结果显示:在这种反应器中,在空气压力脉动的带动下,反应器内的温度和空气湿度也会发生较大幅度的周期性变化,变化的周期和空气压力脉动的周期相同,变化的幅度随着压力脉动的幅度增大而增加;在外界周期刺激的条件下,较未加外界周期刺激斜卧青霉的生物量增加了104% ,二氧化碳的产生总量增加了229%和纤维素酶的产量增加了320 % ,数据显示外界周期刺激不仅增加了菌体的生物量同时增加了其代谢活性。     

Differential neural control of intrarenal blood flow

To test whether renal sympathetic nerve activity (RSNA) can differentially regulate blood flow in the renal medulla (MBF) and cortex (CBF) of pentobarbital sodium-anesthetized rabbits, we electrically stimulated the renal nerves while recording total renal blood flow (RBF), CBF, and MBF. Three stimulation sequences were applied 1) varying amplitude (0.5-8 V), 2) varying frequency (0.5-8 Hz), and 3) a modulated sinusoidal pattern of varying frequency (0. 04-0.72 Hz). Increasing amplitude or frequency of stimulation progressively decreased all flow variables. RBF and CBF responded similarly, but MBF responded less. For example, 0.5-V stimulation decreased CBF by 20 +/- 9%, but MBF fell by only 4 +/- 6%. The amplitude of oscillations in all flow variables was progressively reduced as the frequency of sinusoidal stimulation was increased. An increased amplitude of oscillation was observed at 0.12 and 0.32 Hz in MBF and to a lesser extent RBF, but not CBF. MBF therefore appears to be less sensitive than CBF to the magnitude of RSNA, but it is more able to respond to these higher frequencies of neural stimulation.     

Dominant side in single-leg stance stability during floor oscillations at various frequencies

Background

We investigated lateral dominance in the postural stability of single-leg stance with anteroposterior floor oscillations at various frequencies.

Methods

Thirty adults maintained a single-leg stance on a force platform for 20 seconds per trial. Trials were performed with no oscillation (static condition) and with anteroposterior floor oscillations (2.5-cm amplitude) at six frequencies: 0.25, 0.5, 0.75, 1.0, 1.25 and 1.5 Hz (dynamic condition). A set of three trials was performed on each leg in each oscillation frequency in random order. The mean speed of the center of pressure in the anteroposterior direction (CoP_ap) was calculated as an index of postural stability, and frequency analysis of CoP_ap sway was performed. Footedness for carrying out mobilizing activities was assessed with a questionnaire.

Results

CoP_ap speed exponentially increased as oscillation frequency increased in both legs. The frequency analysis of CoP_ap showed a peak <0.3 Hz at no oscillation. The frequency components at 0.25-Hz oscillation included common components with no oscillation and those at 1.5-Hz oscillation showed the maximum amplitude among all conditions. Postural stability showed no significant difference between left- and right-leg stance at no oscillation and oscillations ≤1.25 Hz, but at 1.5-Hz oscillation was significantly higher in the right-leg stance than in the left-leg stance. For the lateral dominance of postural stability at individual levels, the lateral difference in postural stability at no oscillation was positively correlated with that at 0.25-Hz oscillation (r = 0.51) and negatively correlated with that at 1.5-Hz oscillation (r = -0.53). For 70% of subjects, the dominant side of postural stability was different at no oscillation and 1.5-Hz oscillation. In the subjects with left- or right-side dominance at no oscillation, 94% or 38% changed their dominant side at 1.5-Hz oscillation, with a significant difference between these percentages. In the 1.5-Hz oscillation, 73% of subjects had concordance between the dominant side of postural stability and that of mobilizing footedness.

Conclusion

In static conditions, there was no lateral dominance of stability during single-leg stance. At 1.5-Hz oscillation, the highest frequency, right-side dominance of postural stability was recognized. Functional role in supporting leg may be divided between left and right legs according to the change of balance condition from static to dynamic.     

Effects of deuterium oxide and temperature on heart rate in Drosophila melanogaster

A non-intrusive optical technique has been developed to monitor heartbeat in late third-instar Drosophila larvae. Heartbeat in this insect is an oscillation that is not temperature compensated. Deuterium oxide lengthens the period of a number of high and low frequency oscillators and clocks in a variety of organisms. To determine whether deuterium affects heart rate, flies were raised on proteated and deuterated media and their heartbeat was monitored at four temperatures ranging from 18 to 33 degrees C. The rate of heartbeat increased linearly with increasing temperature, and decreased with increasing concentrations of deuterium. There was a significant interaction between temperature and deuterium: the higher the concentration of deuterium oxide the less temperature-sensitive was the heart rate. Raising temperatures also increased the amount of "noise" in the rhythm: signal-to-noise ratio, which characterizes the amount of power in a rhythmic signal, decreased with increasing temperatures. Deuterium oxide had no effect on signal-to-noise ratio.     

Antiphase oscillations of endothelium and smooth muscle [Ca2+]i in vasomotion of rat mesenteric small arteries

The mechanisms leading to vasomotion in the presence of noradrenaline and inhibitors of the sarcoplasmic/endoplasmic reticulum calcium ATPase were investigated in isolated rat mesenteric small arteries. Isobaric diameter and isometric force were measured together with membrane potential in endothelial cells and smooth muscle cells (SMC). Calcium in the endothelial cells and SMC was imaged with confocal microscopy. In the presence of noradrenaline and cyclopiazonic acid, ryanodine-insensitive oscillations in tone were produced. The frequency was about 1 min(-1) and amplitude about 70% of the maximal tone. The amplitude was reduced by indomethacin and increased with L-NAME. Vasomotion was inhibited by nifedipine and by 40 mM potassium. The frequency was increased and amplitude decreased by removal of the endothelium and by application of charybdotoxin and apamin. The vasomotion was associated with in-phase oscillations of membrane potential in endothelial cells and SMC and oscillations of [Ca2+]i that were in near anti-phase. We suggest a working model for the generation of oscillation based on a membrane oscillator where ion channels in both endothelial cells and SMC interact via a current running between the two cell types through myoendothelial gap junctions, which sets up a near anti-phase oscillation of [Ca2+]i in the two cell types.     

Postural muscle activity patterns during standing at rest and on an oscillating floor.

Postural muscle activity pattern was examined in the eyes-closed state after adequate adaptation to floor anteroposterior oscillation. Twenty-three subjects were grouped almost evenly according to dominance of anterior or posterior postural muscles in the trunk and thigh during quiet stance. In the posterior-dominant group, this dominance was maintained at every frequency in most subjects. In the anterior-dominant group, this dominance was maintained in most subjects at 0.1 and 0.5 Hz but changed to posterior dominance at 1.0 and 1.5 Hz in about half the subjects. Periodicity of muscle activity was evaluated by EMG amplitude spectrum at the floor oscillation frequency. Periodicity of posterior-dominant muscles in the trunk and thigh increased with increasing oscillatory frequency. In the trunk, the periodicity did not differ significantly between posterior-dominant and anterior-dominant groups. However, in the thigh, periodicity was significantly lower in the anterior-dominant muscles. This was considered to be caused by nonperiodic alternating action of the anterior and posterior muscles. In the lower leg, posterior dominance was observed in quiet stance and at all oscillation frequencies. Periodicity of soleus and gastrocnemius increased at higher frequencies and was higher in gastrocnemius than in soleus. The periodicity difference between both muscles decreased with increasing oscillation frequency.     

Actin Microfilament Involved in Regulation of Pacemaking Activity in Cultured Interstitial Cells of Cajal from Murine Intestine

The present study investigated the effect of actin microfilament structure on pacemaker currents and calcium oscillation in cultured murine intestinal interstitial cells of Cajal (ICCs) by whole-cell patch-clamp technique and calcium imaging technique. Cytochalasin B, a disruptor of actin microfilaments, decreased the amplitude and frequency of pacemaker currents from 491.32 ± 160.33 pA and 11.73 ± 0.79 cycles/min to 233.12 ± 92.00 pA and 10.29 ± 0.76 cycles/min. Cytochalasin B also decreased the amplitude and frequency of calcium oscillation from 0.32 ± 0.08 (ΔF/F0) and 2.75 ± 0.17 cycles/min to 0.02 ± 0.01 (ΔF/F0) and 1.20 ± 0.08 cycles/min. Phalloidin, a stabilizer of actin microfilaments, increased the amplitude and frequency of pacemaker currents from 751.79 ± 282.82 pA and 13.93 ± 1.00 cycles/min to 1234.34 ± 607.83 pA and 14.68 ± 1.00 cycles/min. Phalloidin also increased the amplitude and frequency of calcium oscillation from 0.26 ± 0.01 (ΔF/F0) and 2.27 ± 0.18 cycles/min to 0.43 ± 0.03 (ΔF/F0) and 2.87 ± 0.07 cycles/min. 2-Aminoethoxydiphenyl borane (2-APB), an IP₃ receptor blocker, suppressed both pacemaker currents and calcium oscillations. 2-APB also blocked the phalloidin-induced increase in pacemaker currents and calcium oscillation. Ryanodine, an inhibitor of calcium-induced calcium release, did not affect pacemaker current but suppressed calcium oscillations. Ryanodine had no effect on altering phalloidin-induced increases in pacemaker current and calcium oscillation. These results suggest that actin microfilaments regulate pacemaker activity via the IP₃-induced calcium release signaling pathway.     

The control of oscillatory movements of the forearm

The patterns of EMG activity in the biceps and triceps muscles were recorded during horizontal oscillatory movements of the forearm. Subjects showed increased frequency of oscillation as they voluntarily reduced movement amplitude. EMG burst duration was significantly correlated with wavelength of oscillation in every case. In almost half the cases burst intensity was also positively correlated with wavelength. Subjects seemed to be using one or both these methods to control amplitude. A model was developed in three stages which satisfactorily accounted for the data.     

An experimental study on in vivo dynamic mechanical properties of the knees of cats in valgus loading

In these experiments an attempt was made to investigate in vivo dynamic mechanical properties of the knee in valgus loading. Fifteen adult cats were used. Sinusoidal bending oscillation of 0.3, 1.0, 1.6, 3.3, 5.0, 8.6 and 10.0 Hz was transmitted to the knee-leg complex under each pre-bending strain in the valgus direction, and the bending moment was measured. Dynamic mechanical properties of the knee-leg complex were given as a complex bending elastic coefficient. Bending elastic and bending damping coefficient of the knee could be calculated by the least square method. As a first approximation, the knee was represented as a non-linear Kelvin model. Both bending elastic and bending damping coefficient of the knee increased as pre-bending moment was increased, and decreased as amplitude of bending oscillation was increased. Mechanical loss tangent of the knee-leg complex decreased within the range of pre-bending moment larger than 1.0 X 10(6) dyn cm as bending moment was increased, and decreased as amplitude of bending oscillation was increased.     

Modification of intramuscular pH oscillations in the isolated perfused rat heart by different interventions.

Changes in the intramuscular pH oscillations were examined by the use of an antimony electrode upon perfusing the isolated rat heart under different experimental conditions. The pH oscillations were decreased upon perfusing the hearts with Na+- or Ca2+-free medium and increased upon perfusing with K+-free medium. Increasing the temperature of perfusion medium from 25 to 40 degrees C or omitting glucose from the perfusing medium decreased the magnitude of oscillations. On the other hand, complete interruption of the perfusion flow resulted in an increase in the amplitude of pH oscillation. An initial increase followed by a decrease in the pH oscillation was seen when hearts were perfused with medium containing lactic acid at pH 6.6. These results suggest that pH oscillations reflect fluctuations in myocardial metabolism.     

Role of membrane potential in vasomotion of isolated pressurized rat arteries

Vasomotion, the phenomenon of vessel diameter oscillation, regulates blood flow and resistance. The main parameters implicated in vasomotion are particularly the membrane potential and the cytosolic free calcium in smooth muscle cells. In this study, these parameters were measured in rat perfused-pressurized mesenteric artery segments. The application of norepinephrine (NE) caused rhythmic diameter contractions and membrane potential oscillations (amplitude; 5.3 +/- 0.3 mV, frequency; 0.09 +/- 0.01 Hz). Verapamil (1 microM) abolished this vasomotion. During vasomotion, 10(-5) M ouabain (Na(+)-K(+) ATPase inhibitor) decreased the amplitude of the electrical oscillations but not their frequency (amplitude; 3.7 +/- 0.3 mV, frequency; 0.08 +/- 0.002 Hz). Although a high concentration of ouabain (10(-3) M) (which exhibits non-specific effects) abolished both electrical membrane potential oscillations and vasomotion, we conclude that the Na+-K+ ATPase could not be implicated in the generation of the membrane potential oscillations. We conclude that in rat perfused-pressurized mesenteric artery, the slow wave membrane type of potential oscillation by rhythmically gating voltage-dependent calcium channels, is responsible for the oscillation of intracellular calcium and thus vasomotion.     

Effect of the pacing interval and extrasystole on amplitude and damping of diastolic oscillations in the rabbit papillary muscle

Diastolic mechanical oscillations of right ventricular rabbit papillary muscles investigated at 15 degrees C in the perfusion chamber were analysed using a relaxation model with 6 parameters. From this analysis follows that the first diastolic oscillation amplitude plotted against the driving interval ("interval amplitude curve" of the first diastolic oscillation) shows tow maxima thus differing from the shape of the interval strength curve of the preceding driven contraction. It is concluded that the amplitude of diastolic oscillations is less determined by the Ca-amount released from the sarcoplasmic reticulum during the action potential than by the frequency of processes which effect the Ca release and the Ca-sequestration. The damping ratio varies within the range of tested driving intervals (0.36 to 10 s). After an extrasystole damping ratio and period of diastolic oscillation are diminished compared with the values after regularly driven contractions.     

Low-frequency oscillation of sympathetic nerve activity decreases during development of tilt-induced syncope preceding sympathetic withdrawal and bradycardia

Sympathetic activation during orthostatic stress is accompanied by a marked increase in low-frequency (LF, approximately 0.1-Hz) oscillation of sympathetic nerve activity (SNA) when arterial pressure (AP) is well maintained. However, LF oscillation of SNA during development of orthostatic neurally mediated syncope remains unknown. Ten healthy subjects who developed head-up tilt (HUT)-induced syncope and 10 age-matched nonsyncopal controls were studied. Nonstationary time-dependent changes in calf muscle SNA (MSNA, microneurography), R-R interval, and AP (finger photoplethysmography) variability during a 15-min 60 degrees HUT test were assessed using complex demodulation. In both groups, HUT during the first 5 min increased heart rate, magnitude of MSNA, LF and respiratory high-frequency (HF) amplitudes of MSNA variability, and LF and HF amplitudes of AP variability but decreased HF amplitude of R-R interval variability (index of cardiac vagal nerve activity). In the nonsyncopal group, these changes were sustained throughout HUT. In the syncopal group, systolic AP decreased from 100 to 60 s before onset of syncope; LF amplitude of MSNA variability decreased, whereas magnitude of MSNA and LF amplitude of AP variability remained elevated. From 60 s before onset of syncope, MSNA and heart rate decreased, index of cardiac vagal nerve activity increased, and AP further decreased to the level at syncope. LF oscillation of MSNA variability decreased during development of orthostatic neurally mediated syncope, preceding sympathetic withdrawal, bradycardia, and severe hypotension, to the level at syncope.     

Fluttering target detection in Hipposiderid bats

Summary Two species of Hipposiderid bats,Hiposideros speoris andH. lankadiva, which both emit short CF-FM echolocation calls, were trained in a two-alternative forced-choice procedure to discriminate between an oscillating target and a motionless one. Two different targets were used: (1) the membrane of a low-frequency loudspeaker, producing sinusoidal frequency- and amplitude modulations and (2) a small rotating propeller, which produced short acoustical glints. In both casesH. lankadiva learned to discriminate between the oscillating and the motionless target. When the loudspeaker was used, thresholds for minimal modulation depths at different oscillation frequencies were determined. At loudspeaker membrane oscillation frequencies of 10 to 100 Hz the 75% correct thresholds lay between 90 and 300 Hz (Fig. 3).H. speoris could not be trained to react to the moving membrane, even at very high oscillation amplitudes. When the rotating propeller was the positive target, however,H. speoris learned very quickly to discriminate it from a motionless one. By decreasing the rotation speed it was possible to measure the minimal detectable glint-frequency for each bat. It lay at 67, 44, and just under 15 glints/s for the three specimens (Fig. 4). During the discrimination task both bat species increased their duty-cycle just prior to a decision by emitting long sequences of echolocation calls with short inter-pulse intervals. The duration of individual pulses remained relatively constant (Figs. 6 and 7). Possible mechanisms for discrimination of the oscillating targets are discussed and the importance of glints in the echoes for fluttering target detection is emphasized.Abbreviations A₀ amplitude of loudspeaker membrane oscillation - AM amplitude modulation - CF constant frequency - DC duty-cycle - FM frequency modulation - f _max maximal frequency modulation depth in echo - PD pulse duration - PI pulse interval - f _osc oscillation frequency