Biomedical subjects

B J Simon

Publications and source records attributed to B J Simon.

33 records · Page 2Linked to original sources

PubMed1991

Biochemical aspects of H(+)-ATPase in renal proximal tubules: inhibition by N,N'-dicyclohexylcarbodiimide, N-ethylmaleimide, and bafilomycin.

Brush-border membranes from rat kidney cortex are transiently exposed to cholate to reorient ATP-driven H+ pumps to the outside of the vesicles. The carboxyl group reagent, N,N'-dicyclohexylcarbodiimide (DCCD), inhibits ATP-driven H+ uptake into cholate-pretreated vesicles irreversibly. Complete inhibition requires treatment of vesicles with 0.2 mM DCCD for greater than or equal to 15 min. ATP and ADP do not protect the H+ pump from inactivation suggesting that DCCD modifies pump subunits involved in H+ translocation, but not those related to ATP hydrolysis. With [14C]DCCD a 16 kDa protein is strongly labeled in brush-border and endosomal membranes, but not in basolateral membranes. Molecular mass of this protein and distribution similar to H(+)-ATPases suggest a role as H(+)-conducting subunit of the H+ pumps. The SH-group reagent, N-ethylmaleimide (NEM), also inhibits ATP-driven H+ uptake irreversibly. As opposed to DCCD, ATP and ADP protect the pump from irreversible inhibition indicating that NEM modifies SH-groups in the proximity of ATP hydrolysis sites. Finally, 15 nM of a potent inhibitor of vacuolar ATPases, bafilomycin B1, abolishes ATP-driven H+ uptake. Inactivation by DCCD and NEM, labeling of 16 kDa subunits by [14C]DCCD, and high sensitivity to bafilomycin indicate that the H+ pump (H(+)-ATPase) in rat renal brush-border membranes belongs to the class of vacuolar ATPases. Bafilomycin may prove a valuable tool for specific inhibition of the renal H(+)-ATPase in future studies.

Adenosine Triphosphate↗

PubMed1990

The ATP-binding site of the human placental H+ pump contains essential tyrosyl residues.

Transient exposure of human placental brush-border membrane vesicles to cholate reorients the ATP-driven H+ pump, enabling the pump to transport H+ into the vesicles upon addition of ATP to the external medium. H+ uptake can be measured in these vesicles by following the decrease in the absorbance of acridine orange, a delta pH indicator. We investigated the role of tyrosyl residues in the catalytic function of the H+ pump by studying the effects of tyrosyl group specific reagents on ATP-driven H+ uptake in cholate-pretreated membrane vesicles. The reagents tested were 7-chloro-4-nitro-2,1,3-benzoxadiazole (NBD-Cl), N-acetylimidazole, tetranitromethane, and p-nitrobenzenesulfonyl fluoride. Treatment of the membrane vesicles with these reagents resulted in the inhibition of the ATP-driven H+ uptake, and the inhibitory potency was in the following order: NBD-Cl greater than tetranitromethane greater than p-nitrobenzenesulfonyl fluoride greater than N-acetylimidazole. The inhibition of the H+ pump by NBD-Cl was reversible by 2-mercaptoethanol, and the inhibition by N-acetylimidazole was reversible by hydroxylamine. Since these reagents are not absolutely specific for tyrosyl groups and can also react with thiol groups, we studied the interaction of N-acetylimidazole with the H+ pump whose triol groups were masked by reaction with p-(chloromercuri)benzenesulfonate. The SH-masked pump was totally inactive, but the activity could be restored by dithiothreitol. On the contrary, the activity of the SH-masked H+ pump which was subsequently treated with N-acetylimidazole could not be restored by dithiothreitol, suggesting that thiol groups were not involved in the inhibition of the H+ pump by N-acetylimidazole.(ABSTRACT TRUNCATED AT 250 WORDS)

4-Chloro-7-nitrobenzofurazan↗

PubMed1990

An essential role for vicinal dithiol groups in the catalytic activity of the human placental Na(+)-H+ exchanger.

We examined the effects of phenylarsine oxide, a reagent specific for vicinal dithiol groups, on the catalytic activities, Na+ influx and H+ efflux, of the human placental Na(+)-H+ exchanger. Treatment of the placental brush-border membrane vesicles with the reagent markedly inhibited both the activities. The inhibition was partially reversible by dithiols. The effect of phenylarsine oxide was to reduce the maximal velocity of the exchanger without influencing its affinity for Na+. The exchanger was partially protected from this inhibition by amiloride but not by cimetidine even though both these compounds interacted with the Na(+)-binding site. The data demonstrate that vicinal dithiol groups are essential for the catalytic function of the placental Na(+)-H+ exchanger and that the critical dithiol groups are located at a site distinct from the Na(+)-binding site.

Amiloride↗

PubMed1990

Characterization of inside-out oriented H(+)-ATPases in cholate-pretreated renal brush-border membrane vesicles.

Exposure of porcine renal brush-border membrane vesicles to 1.2% cholate and subsequent detergent removal by dialysis reorients almost all N-ethylmaleimide (NEM)-sensitive ATPases from the vesicle inside to the outside. ATP addition to cholate-pretreated, but not to intact, vesicles causes H+ uptake as visualized by the delta pH indicator, acridine orange. The reoriented H(+)-pump is electrogenic because permeant extravesicular anions or intravesicular K+ plus valinomycin enhance H+ transport. ATP stimulates H+ uptake with an apparent Km of 93 microM. Support of H+ uptake and Pi liberation by ATP greater than GTP approximately ITP greater than UTP indicates a preference for ATP and utilization of other nucleotides at lower efficiency. ADP is a potent, competitive inhibitor of ATP-driven H+ uptake (Ki, 24 microM), Mg2+ and Mn2+ support ATP-driven H+ uptake, but Ca2+, Ba2+, and Zn2+ do not, 1 mM Zn2+ inhibits MgATP-driven H+ transport completely. NEM-sensitive Pi liberation is stimulated by Mg2+ and Mg2+ and, unlike H+ uptake, also by Ca2+ suggesting Ca2(+)-dependent ATP hydrolysis unrelated to H+ transport. The inside-out oriented H(+)-pump is relatively insensitive toward oligomycin, azide, N,N'-dicyclohexylcarbodiimide (DCCD) and vanadate, but efficiently inhibited by NEM (apparent Ki, 0.77 microM), and 4-chloro-7-nitro-benzoxa-1,3-diazole (NBD-Cl; apparent Ki, 0.39 microM). Taken together, the H(+)-ATPase of proximal tubular brush-border membranes exhibits characteristics very similar to those of "vacuolar type" (V-type) H(+)-ATPases. Hence, V-type H(+)-ATPases occur not only in intracellular organelles but also in specialized plasma membrane areas.

Acridine Orange↗

PubMed1990

Effects of caffeine on calcium release from the sarcoplasmic reticulum in frog skeletal muscle fibres.

1. Resting myoplasmic [Ca2+] and [Ca2+] transients (delta [Ca2+]) were monitored using Fura-2 fluorescence and Antipyrylazo III absorbance signals from voltage-clamped segments of cut frog skeletal muscle fibres in the presence and absence of 0.5 mM-caffeine. The rate of release (Rrel) of calcium from the sarcoplasmic reticulum was calculated from delta [Ca2+]. 2. delta [Ca2+] and Rrel were increased in caffeine for all pulses. The decline of delta [Ca2+] was slower after a given pulse in caffeine than without caffeine. Resting [Ca2+] was slightly elevated in caffeine. 3. The voltage dependence of the peak value of Rrel and of the steady level of Rrel at the end of a 60-120 ms pulse were both shifted towards more negative voltages in caffeine. For relatively small pulses the voltage at which a given release waveform was observed was also shifted to more negative voltages. 4. Intramembrane charge movements measured in the same fibres in which the above changes in Rrel were observed showed no significant changes in caffeine. 5. In caffeine calcium release continued for many milliseconds after the end of a short (10 ms) pulse. Continued release after a pulse was not observed without caffeine and was probably due to positive feedback of elevated [Ca2+] on calcium release resulting from calcium-induced calcium release in caffeine. 6. Intramembrane charge movements after short pulses showed no change in caffeine that could account for the continued calcium release after the pulse. 7. Continued release after short pulses in caffeine decreased as the pulse duration was increased and was absent for pulses of 60 ms or longer. Rrel also inactivated during such pulses. 8. Relatively large and long conditioning pulses in caffeine suppressed both the peak Rrel and the continued release after short pulses. Peak release and continued release after short pulses recovered in parallel with increasing recovery time following suppression by a conditioning pulse in caffeine. 9. These results indicate that in the presence of caffeine, charge movement and calcium-induced calcium release both contribute significantly to the activation of sarcoplasmic reticulum calcium release during fibre depolarization. Release activated by either mechanism appears to be inactivated by calcium-dependent inactivation. A significant contribution of calcium-induced calcium release during depolarization in the absence of caffeine is not ruled out by present observations.

Animals↗

PubMed1989

Caffeine slows turn-off of calcium release in voltage clamped skeletal muscle fibers.

Myoplasmic free calcium transients delta [Ca2+] were monitored with the calcium indicators antipyrylazo III and fura-2 in voltage clamped cut frog skeletal muscle fibers, in the presence and absence of 0.5 mM caffeine. Without caffeine delta [Ca2+] began to decline within a few milliseconds of fiber repolarization for pulses of all durations. In caffeine delta [Ca2+] continued to rise for 10-60 ms after 10 or 20 ms depolarizing pulses, indicating that the release of calcium from the sarcoplasmic reticulum (SR) continued well after repolarization of transverse tubular (TT) membranes in the presence of caffeine. Caffeine also increased the peak amplitude of delta [Ca2+] for all pulses and slowed the decline of delta [Ca2+] after pulses of all durations. The rate of calcium release from the SR calculated from delta [Ca2+] showed that for 10 ms pulses in caffeine release did not turn off abruptly on repolarization but instead declined to zero with a time constant essentially the same as the time constant for inactivation of SR calcium release during depolarizing pulses in the presence or absence of caffeine. The observed loss of TT membrane potential control of SR calcium release in the presence of caffeine suggests the appearance of a significant component of cytosolic Ca2+-induced calcium release in caffeine.

Animals↗

PubMed1988

Time course of activation of calcium release from sarcoplasmic reticulum in skeletal muscle.

Myoplasmic free calcium transients were measured with antipyrylazo III in voltage clamped segments of frog skeletal muscle fibers and were used to calculate the rate of release (Rrel) of calcium from the sarcoplasmic reticulum. Intramembrane charge movement was measured for the same pulses in the same fibers. During a depolarizing pulse Rrel rose to an early peak and then decayed relatively rapidly but incompletely due to calcium-dependent inactivation (Schneider M.F., and B.J. Simon. 1988. J. Physiol. (Lond.). 405:727-745). Two approaches were used to determine release activation independent of the effects of inactivation: (a) a mathematical correction based on the assumption that inactivation was a process occurring in parallel with and independently of activation; (b) an experimental procedure in which release was maximally inactivated by a large short prepulse and then the remaining noninactivatable component of release was monitored during a subsequent test pulse. Both procedures gave the same time course of activation of release. Release activation paralleled the time course of intramembrane charge movement but was delayed by a few milliseconds.

Animals↗

PubMed1988

Simultaneous recording of calcium transients in skeletal muscle using high- and low-affinity calcium indicators.

To monitor cytosolic [Ca2+] over a wide range of concentrations in functioning skeletal muscle cells, we have used simultaneously the rapid but relatively low affinity calcium indicator antipyrylazo III (AP III) and the slower but higher affinity indicator fura-2 in single frog twitch fibers cut at both ends and voltage clamped with a double vaseline gap system. When both dyes were added to the end pool solution the cytosolic fura-2 concentration reached a steady level equal to the end pool concentration within approximately 2.5 h, a time when the AP III concentration was still increasing. For depolarizing pulses of increasing amplitude, the fura-2 fluorescence signal approached saturation when the simultaneously recorded AP III absorbance change was far from saturation. Comparison of simultaneously recorded fura-2 and AP III signals indicated that the mean values of the on and off rate constants for calcium binding to fura-2 in 18 muscle fibers were 1.49 x 10(8) M-1 s-1 and 11.9 s-1, respectively (mean KD = 89 nM), if all AP III in the fiber is assumed to behave as in calibrating solution and to be in instantaneous equilibrium with [Ca2+]. [Ca2+] transients calculated from the fura-2 signals using these rate constants were consistent with the [Ca2+] transients calculated from the AP III signals. Resting [Ca2+] or small changes in [Ca2+] which could not be reliably monitored with AP III could be monitored with fura-2 with little or no interference from changes in [Mg2+] or from intrinsic signals. The fura-2 signal was also less sensitive to movement artifacts than the AP III signal. After a [Ca2+] transient the fura-2 signal demonstrated a relatively small elevation of [Ca2+] that was maintained for many seconds.

Animals↗

PubMed1988

Inactivation of calcium release from the sarcoplasmic reticulum in frog skeletal muscle.

1. The rate of calcium release (Rrel) from the sarcoplasmic reticulum (SR) in voltage clamped segments of frog skeletal muscle fibres was calculated from myoplasmic free calcium transients (delta[Ca2+]) measured with the calcium indicator dye Antipyrylazo III. 2. During a 100-200 ms depolarizing pulse Rrel reached an early peak and then declined markedly. The time course and extent of decline of Rrel were independent of membrane potential over a range of potentials where release activation varied severalfold. 3. For test pulses applied shortly after relatively large or long conditioning pulses Rrel completely lacked the early peak. The peak gradually recovered as the interval between the conditioning and test pulses was increased to 1 s. 4. A latency was often observed before the start of recovery of the peak in Rrel. The latency appeared to be correlated with the time for delta[Ca2+] to fall below a certain level, indicating that recovery of the peak might represent reversal of a calcium-dependent process. It was therefore proposed that the rapid decline in Rrel during a pulse was due to calcium-dependent inactivation of the SR calcium release channels. 5. Inactivation continued to develop during the interval between a relatively large 20 ms conditioning pulse and a test pulse applied 20 ms later. This was as expected for calcium-dependent inactivation of SR calcium release because of the elevated [Ca2+] between the conditioning and test pulses. It was not as expected for external membrane potential-dependent inactivation. 6. Small steady elevations in [Ca2+] due to relatively small 200 ms conditioning pulses produced marked inactivation of Rrel, indicating an apparent dissociation constant for calcium-dependent inactivation only slightly above resting [Ca2+]. 7. All observations could be well simulated by a two-step model for inactivation in which myoplasmic free calcium equilibrates rapidly with a high-affinity calcium receptor on the release channel and then the calcium-receptor complex undergoes a slower conformational change to the inactivated state of the channel. 8. An alternative model in which calcium binds to a soluble receptor (e.g. free calmodulin) and then the calcium-receptor complex binds to and directly inactivates the channel was shown to be formally identical to the preceding model. Either model could closely simulate all observations.

Animals↗

PubMed1987

Depletion of calcium from the sarcoplasmic reticulum during calcium release in frog skeletal muscle.

1. Free intracellular calcium transients (delta[Ca2+] were monitored in cut segments of frog skeletal muscle fibres voltage clamped in a double Vaseline-gap chamber and stretched to sarcomere lengths that eliminated fibre movement. The measured calcium transients were used to calculate the rate of calcium release from the sarcoplasmic reticulum (s.r.) as previously described (Melzer, Rios & Schneider, 1984, 1987). 2. Conditioning pulses were found to suppress the rate of calcium release in test pulses applied after the conditioning pulse. Various combinations of conditioning and test pulses were used to investigate the basis of the suppression of calcium release by the conditioning pulse. 3. Using a constant test pulse applied at varying intervals after a constant conditioning pulse, recovery from suppression of release was found to occur in two phases. During the fast phase of recovery, which was completed within about 1 s, the rate of calcium release was smaller and had a different wave form than the unconditioned control release. The early peak in release that is characteristic of the control release wave form was absent or depressed. During the slow phase of recovery, which required about 1 min for completion, the release wave form was the same as control but was simply scaled down compared to the control. 4. Conditioning pulses also slowed the rate of decay of delta[Ca2+] after a constant test pulse, probably due to an increased occupancy by calcium of slowly equilibrating myoplasmic sites that bind some of the calcium released by the conditioning pulse. Since calcium binding to these sites contributes to the decay of delta[Ca2+], their increased occupancy would slow the decay of delta[Ca2+] following the test pulse. This effect was used to estimate the calcium occupancy of the slowly equilibrating sites. 5. Comparison of the time course of the slow recovery from suppression of release following a constant conditioning pulse with the time course of the loss of calcium from the slowly equilibrating myoplasmic calcium binding sites indicated that the two processes occurred in parallel. 6. Using a set 1 s recovery period and a constant test pulse but varying the amplitude and/or duration of the conditioning pulse, the degree of slowly recovering suppression of release was found to be directly related to the amount of calcium remaining outside of the s.r. at the start of the test pulse. 7. Points 3, 5 and 6 above indicate that the slow recovery from suppression of release may be due to slow recovery from depletion of calcium from the s.r.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

PubMed1986

Intramembrane charge movement and calcium release in frog skeletal muscle.

Intramembrane charge movement and myoplasmic free calcium transients (delta[Ca2+]) were monitored in voltage-clamped segments of isolated frog muscle fibres cut at both ends and mounted in a double Vaseline-gap chamber. The fibres were stretched to sarcomere lengths of 3.5-4.6 micron to minimize mechanical movement and the related optical artifacts. The over-all calcium removal capability of each fibre was characterized by analysing the decay of delta[Ca2+] following pulses of several different amplitudes and durations. The rate of sarcoplasmic reticulum (s.r.) calcium release was then calculated for each delta[Ca2+] using the calcium removal properties determined for that fibre. The calculated calcium release wave form reached a relatively early peak and then declined appreciably during a 100-150 ms depolarizing pulse. The voltage dependence of the peak rate of calcium release was steeper and was centred at more positive membrane potentials than the steady-state voltage dependence of charge movement in the same fibres. A considerable fraction of the total intramembrane charge was moved at potentials at which delta[Ca2+] and calcium release were only a few per cent of maximum. This 'subthreshold' charge may correspond to charge moved in preliminary transitions that precede a final charge transition that activates release. A 'stepped on' pulse protocol was used to experimentally separate the subthreshold charge movement from the charge movement of the final transitions that may control calcium release. The stepped on pulse consisted of a set 50 ms pre-pulse to a potential just at or below the potential for detectable delta[Ca2+] followed immediately by a test pulse of varying amplitude and duration. For a wide range of test pulse amplitudes and durations in the stepped on protocol the peak rate of calcium release was linearly related to the charge movement during the test pulse. This result points to a tight control of activation of s.r. calcium release by intramembrane charge movement. The voltage dependence of both charge movement and of the rate of calcium release could be fitted simultaneously with a three-state, two-transition sequential model in which charge moves in both transitions but only the final transition activates s.r. calcium release. A model with three identical and independent charged gating particles per channel gave an equally good fit to the data. Both models closely fit the charge movement and release data except within about 10 mV of the voltage at which release became detectable, where release varied more steeply with membrane potential than predicted by either model.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

PubMed1985

Slow charge movement in mammalian skeletal muscle.

Voltage-dependent charge movements were measured in the rat omohyoid muscle with the three-microelectrode voltage-clamp technique. Contraction was abolished with hypertonic sucrose. The standard (ON-OFF) protocol for eliciting charge movements was to depolarize the fiber from -90 mV to a variable test potential (V) and then repolarize the fiber to -90 mV. The quantity of charge moved saturated at test potentials of approximately 0 mV. The steady state dependence of the amount of charge that moves as a function of test potential could be well fitted by the Boltzmann relation: Q = Qmax/(1 + exp[-(V - V)/k]), where Qmax is the maximum charge that can be moved, V is the potential at which half the charge moves, and k is a constant. At 15 degrees C, these values were Qmax = 28.5 nC/microF, V = -34.2 mV, and k = 8.7 mV. Qmax, k, and V exhibited little temperature dependence over the range 7-25 degrees C. "Stepped OFF" charge movements were elicited by depolarizing the fiber from -90 mV to a fixed conditioning level that moved nearly all the mobile charge (0 mV), and then repolarizing the fiber to varying test potentials. The sum of the charge that moved when the fiber was depolarized directly from -90 mV to a given test potential and the stepped OFF charge that moved when the fiber was repolarized to the same test potential had at all test potentials a value close to Qmax for that fiber. In nearly all cases, the decay phase of ON, OFF, and stepped OFF charge movements could be well fitted with a single exponential. The time constant, tau decay, for an ON charge movement at a given test potential was comparable to tau decay for a stepped OFF charge movement at the same test potential. Tau decay had a bell-shaped dependence on membrane potential: it was slowest at a potential near V (the midpoint of the steady state charge distribution) and became symmetrically faster on either side of this potential. Raising the temperature from 7 to 15 degrees C caused tau decay to become faster by about the same proportion at all potentials, with a Q10 averaging 2.16. Raising the temperature from 15 to 25 degrees C caused tau decay to become faster at potentials near V, but not at potentials farther away.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1985

The influence of transverse tubular delays on the kinetics of charge movement in mammalian skeletal muscle.

A model was developed to describe the kinetics of slow, voltage-dependent charge movement in the rat omohyoid muscle. To represent the electrically distributed nature of the transverse tubular system (t-system), we followed an approach similar to that described by Adrian and Peachey (1973 J. Physiol. [Lond.]. 235:103), and approximated the fiber with 12 concentric cylindrical shells. Incorporated into each shell were capacitative and conductive elements that represented the passive electrical properties of the t-system, and an element representing the mobile charge. The charge was assumed to obey a two-state scheme, in which the redistribution of charge is governed by a first-order reaction, and the rate constants linking the two states were assumed to depend on potential according to the constant field expression. The predictions of this "distributed two-state model" were compared with charge movements experimentally measured in individual fibers. For this comparison, first, the passive electrical parameters of the model were adjusted to fit the experimental linear capacity transient. Next, the Boltzmann expression was fitted to the steady state Q vs. V data of the fiber, thereby constraining the voltage dependence of the rate constants, but not their absolute magnitude. The absolute magnitude was determined by fitting the theory to an experimental charge movement at a single test potential, which in turn constrained the fits at all other test potentials. The distributed two-state model well described the rising and falling phases of ON, OFF, and stepped OFF charge movements at temperatures ranging from 3 to 25 degrees C. We thus conclude that tubular delays are sufficient to account for the rounded rising phase of experimental charge movements, and that it is unnecessary to postulate higher-order reaction schemes for the underlying charge redistribution.

Animals↗

PubMed1983

Charge movement in a fast twitch skeletal muscle from rat.

Voltage-dependent charge movement in the rat omohyoid muscle was investigated using the three microelectrode voltage clamp technique. The charge that moved during a depolarization from the holding potential (-90 mV) to the test potential, V, increased with increasing V, saturating around 0 mV. The charge vs. voltage relationship was well fitted by Q = Q(max)/{1 + exp[-(V - V)/k]}, with Q(max) = 28.5 nC/muF, V = -34.2 mV, and k = 8.7 mV. Repolarization of the fiber from the test potential back to the holding potential caused an equal but opposite amount of charge to move. The kinetics of ON charge movement could be well described by a model developed for frog muscle by Horowicz and Schneider (1981b), which suggests that rat and frog charge movements are similar. This model failed to describe the kinetics of OFF charge movement for steps in potential from 0 mV to test potentials of -10 to -90 mV. OFF-charge movement rose to a peak more slowly and decayed more slowly than predicted by the theory.

Animals↗

PubMedDate not supplied

The effect of varied electrical current densities on lumbar spinal fusions in dogs.

BACKGROUND CONTEXT: The use of electrical stimulation has been shown to increase the rate of successful spinal fusions. It is possible that increasing the current density of these stimulators may increase the speed and success rate of these fusions. PURPOSE: This study evaluated the effects of varied current densities on the speed and success rate of spinal fusion in dogs. STUDY DESIGN/SETTING: Three different current densities (0.83 microA/cm, 4 microA/cm and 10 microA/cm) were used to stimulate spinal fusions in a canine model over a 12-week period. OUTCOME MEASURES: Radiographic and histologic assessments were used to determine the degree of facet fusions at each time period. METHODS: Fifteen dogs underwent spinal facet fusion bilaterally at the level of L1-2 and L4-5. Each fusion site was electrically stimulated using one of three current densities. At 6, 9 and 12 weeks, the specimens were evaluated using high-resolution radiography and routine histology. The fusion masses were graded and then statistically evaluated. RESULTS: The results demonstrate a dose response of fusion mass scores to increasing current density. The highest current density (10 microA/cm) demonstrated a statistically higher fusion scores than the lowest currently density (0.83 microA/cm) at 6 weeks and statistically greater than both the middle (4 microA/cm) and lowest (0.83 microA/cm) current densities at 9 weeks. No differences were noted at 12 weeks, because all groups showed complete fusion. CONCLUSIONS: This controlled study suggests that speed of fusion may be further improved by increasing the current density of the electrical stimulation.

Animals↗