Firing properties of muscle spindles supplying the intrinsic foot muscles of humans in unloaded and freestanding conditions.

We recently developed an approach for recording from muscle spindles in the intrinsic muscles of the foot in freestanding humans by inserting a tungsten microelectrode into the posterior tibial nerve behind the medial malleolus of the ankle. Here we characterize the behavior of muscle spindles in the small muscles of the foot in 1) seated subjects with the leg horizontal and the foot naturally plantarflexed and 2) standing subjects. In the first study, recordings were made from 26 muscle spindle afferents located within flexor digiti minimi brevis ( n = 4), abductor digiti minimi ( n = 3), quadratus plantae ( n = 3), plantar interossei ( n = 4), flexor digitorum brevis ( n = 3), dorsal interossei ( n = 2), and lumbricals ( n = 2), with one each supplying abductor hallucis, adductor hallucis, and flexor hallucis brevis. The identity of another two muscle afferents was unknown. The majority of the units were silent at rest, only seven (27%) being spontaneously active. Because of the anatomic constraints of the foot, some spindles supplying muscles acting on the toes responded to movements of one or more digits. In the second study, 12 muscle spindle afferents were examined during standing. The ongoing discharge of eight spindle afferents covaried with changes in the center of pressure during postural sway. We conclude that the majority of spindle endings in the small muscles of the foot are silent at rest, which may allow them to encode changes in conformation of the foot when it is loaded during standing. Moreover, these muscle spindle afferents can provide useful proprioceptive information during standing and postural sway. NEW & NOTEWORTHY We have characterized the firing properties of muscle spindles in the intrinsic muscles of the human foot for the first time. The majority of the spindle endings are silent in seated subjects, and most fire tonically during standing, their discharge covarying with center of pressure during postural sway. We conclude that spindle endings in the intrinsic muscles of the foot provide useful proprioceptive information during free standing.

Microneurography from the posterior tibial nerve: a novel method of recording activity from the foot in freely standing humans.

The posterior tibial nerve, located behind the medial malleolus of the ankle, supplies the intrinsic muscles of the foot and most of the skin of the sole. We describe a novel approach for recording from this nerve via a percutaneously inserted tungsten microelectrode and provide examples of recordings from presumed muscle spindle endings recorded in freely standing human subjects. The fact that the angular excursions of the ankle joint are small as the foot is loaded during the transition from the seated position to standing means that one can obtain stable recordings of neural traffic in unloaded, loaded, and freely standing conditions. We conclude that this novel approach will allow studies that will increase our understanding of the roles of muscle and cutaneous afferents in the foot in the control of upright posture. NEW & NOTEWORTHY We have performed the first microneurographic studies from the posterior tibial nerve at the ankle. Stability of the recording site allows one to record from muscle spindles in the intrinsic muscles of the foot as well as from cutaneous mechanoreceptors in the sole of the foot during the transition from seated to standing. This novel approach opens up new opportunities for studying the roles of muscle and cutaneous afferents in the foot in the control of upright stance.

The vestibular system does not modulate fusimotor drive to muscle spindles in relaxed leg muscles of subjects in a near-vertical position.

It has been shown that sinusoidal galvanic vestibular stimulation (sGVS) has no effect on the firing of spontaneously active muscle spindles in either relaxed or voluntarily contracting human leg muscles. However, all previous studies have been conducted on subjects in a seated position. Given that independent vestibular control of muscle spindle firing would be more valuable during postural threat, we tested the hypothesis that this modulation would become apparent for subjects in a near-vertical position. Unitary recordings were made from 18 muscle spindle afferents via tungsten microelectrodes inserted percutaneously into the common peroneal nerve of awake human subjects laying supine on a motorized tilt table. All recorded spindle afferents were spontaneously active at rest, and each increased its firing rate during a weak static contraction. Sinusoidal bipolar binaural galvanic vestibular stimulation (±2 mA, 100 cycles) was applied to the mastoid processes at 0.8 Hz. This continuous stimulation produced a sustained illusion of "rocking in a boat" or "swinging in a hammock." The subject was then moved into a near-vertical position (75°), and the stimulation repeated. Despite robust vestibular illusions, none of the fusimotor-driven spindles exhibited phase-locked modulation of firing during sinusoidal GVS in either position. We conclude that this dynamic vestibular stimulus was insufficient to modulate the firing of fusimotor neurons in the near-vertical position. However, this does not mean that the vestibular system cannot modulate the sensitivity of muscle spindles via fusimotor neurons in free unsupported standing, when reliance on proprioceptive feedback is higher.

Neck proprioceptors contribute to the modulation of muscle sympathetic nerve activity to the lower limbs of humans.

Several different strategies have now been used to demonstrate that the vestibular system can modulate muscle sympathetic nerve activity (MSNA) in humans and thereby contribute to the regulation of blood pressure during changes in posture. However, it remains to be determined how the brain differentiates between head-only movements that do not require changes in vasomotor tone in the lower limbs from body movements that do require vasomotor changes. We tested the hypothesis that neck movements modulate MSNA in the lower limbs of humans. MSNA was recorded in 10 supine young adult subjects, at rest, during sinusoidal stretching of neck muscles (100 cycles, 35° peak to peak at 0.37 ± 0.02 Hz) and during a ramp-and-hold (17.5° for 54 ± 9 s) static neck muscle stretch, while their heads were held fixed in space. Cross-correlation analysis revealed cyclical modulation of MSNA during sinusoidal neck muscle stretch (modulation index 45.4 ± 5.3 %), which was significantly less than the cardiac modulation of MSNA at rest (78.7 ± 4.2 %). Interestingly, cardiac modulation decreased significantly during sinusoidal neck displacement (63.0 ± 9.3 %). By contrast, there was no significant difference in MSNA activity during static ramp-and-hold displacements of the neck to the right or left compared with that with the head and neck aligned. These data suggest that dynamic, but not static, neck movements can modulate MSNA, presumably via projections of muscle spindle afferents to the vestibular nuclei, and may thus contribute to the regulation of blood pressure during orthostatic challenges.

Stripping voltammetric quantification of the anti-diabetic drug glipizide in bulk form and pharmaceutical formulation.

The electrochemical behavior of glipizide at the hanging mercury drop electrode (HMDE) was studied in B-R universal buffers of pH 1.7-11. The voltammograms exhibited a well-defined 4-electron irreversible cathodic peak which attributed to reduction of the two C=N of the pyrazine ring of glipizide molecule. Glipizide was found to has an interfacial adsorptive character onto the mercury electrode surface. A monolayer surface coverage of 1.02x10(-10)mol cm(-2) was estimated and hence each adsorbed glipizide molecule occupied an area of 1.63 nm(2) onto the mercury electrode surface. A simple and precise square-wave adsorptive cathodic stripping (SWAdCS) voltammetric procedure was described for quantification of bulk glipizide with a limit of detection of 1.5x10(-10)M and a limit of quantitation of 5x10(-10)M. The proposed procedure was successfully applied for quantitation of glipizide in its pharmaceutical formulation (Minidiab tablets) without interference from excipients.

Voltammetric studies on the antibiotic drug cefoperazone quantification and pharmacokinetic studies.

A fully validated simple, sensitive and selective square-wave stripping voltammetry procedure was described for the trace quantification of cefoperazone in bulk form, formulations and human serum/plasma. The procedure was based on reduction of the adsorbed drug onto a hanging mercury drop electrode. The procedural conditions were optimized as: frequency=60Hz, scan increment=8mV, pulse amplitude=25mV, preconcentration potential=-0.3V (versus Ag/Ag/KCl(s)), preconcentration duration=60-150s and an acetate buffer of pH 4.2 as a supporting electrolyte. A limit of detection of 4.5x10(-10)M and a limit of quantitation of 1.5x10(-9)M bulk cefoperazone were achieved following preconcentration of the drug onto the hanging mercury drop electrode for 150s. The proposed square-wave adsorptive cathodic stripping voltammetric procedure was successfully applied for trace quantification of cefoperazone in human serum and plasma. The achieved limits of detection and quantitation of the drug in human serum were 6x10(-10)M (0.375ngml(-1)) and 2x10(-9)M (1.250ngml(-1)), respectively. The pharmacokinetic parameters of cefoperazone in plasma of hospitalized volunteers were successfully estimated.

Adsorptive stripping voltammetric quantification of the antipsychotic drug clozapine in bulk form, pharmaceutical formulation and human serum at a mercury electrode.

Using the cyclic voltammetry technique and the Britton-Robinson buffer (pH 2-10) as a supporting electrolyte, clozapine was found to reduce at the hanging mercury drop electrode in a single two-electron irreversible step corresponding to reduction of the azomethine group of the seven-member heterocyclic ring. Based on the interfacial adsorptive character of clozapine onto the hanging mercury drop electrode, a validated square-wave adsorptive cathodic stripping (SWAdCS) voltammetric procedure was described for the quantification of bulk clozapine with limits of detection and quantitation of 4.5 x 10(-10) and 1.5 x 10(-9) M, respectively. The proposed procedure was successfully applied to the quantification of the drug in pharmaceutical formulation (Leponex) and human serum without the necessity for samples' pretreatment or any time-consuming extraction or evaporation steps prior to the analysis. The limits of detection and quantitation of clozapine in spiked human serum were found to be 1 x 10(-9) and 3.3 x 10(-9) M, respectively. The proposed procedure for quantification of clozapine in bulk form, tablets and human serum has the advantage of being simple, rapid, sensitive, precise and inexpensive compared to most of the reported methods.

Behavior and quantification studies of amiloride drug using cyclic and square-wave adsorptive stripping voltammetry at a mercury electrode.

The cyclic voltammograms of amiloride at the hanging mercury drop electrode showed a single well-defined four-electron irreversible cathodic peak in Britton-Robinson (B-R) buffer of pH 2. At higher pH values (pH > or =3), two irreversible cathodic peaks corresponding to the transfer of four (first peak) and two (second peak) electrons, were obtained The peak potentials were shifted to more negative values on the increase of pH of the medium, implying the involvement of protons in the electrode reaction and that the proton-transfer reaction precedes the proper electrode process. The 4-electron single peak (pH 2) or the first peak (pH > or = 3) may be attributed to the cleavage of the -CH=NH double bond of the N-imidino amide group with the release of NH(3) molecule. While the second peak may be due to the saturation of the C?O double bond of the carboxamide moiety. Based on the interfacial adsorptive character of the drug onto the mercury electrode surface, a simple, sensitive and low cost square-wave adsorptive cathodic stripping (SWAdCS) voltammetric procedure was optimized for analysis of the drug. The optimal operational conditions of the proposed procedure were: accumulation potential E(acc)= -0.7 V, accumulation time t(acc)= 60-65s, scan increment= 10 mV, pulse-amplitude = 50-60 mV, frequency =120 Hz using a B-R buffer of pH 8 as a supporting electrolyte. The linear concentration range was found to be 2 x 10(-9) to 2 x 10(-7) M amiloride with limits of detection (LOD) and quantitation (LOQ) of 1.9 x 10(-10) and 6.3 x 10(-10) M, respectively. The procedure was successfully applied for determination of amiloride in pharmaceutical formulation and spiked in human serum. The LOD and LOQ of amiloride spiked in human serum were 5.7 x 10(-10) and 1.9 x 10(-9) M amiloride, respectively. The procedure did not require sample pretreatment or any time-consuming extraction or evaporation steps, other than deproteinization and then centrifugal separation of protein from serum sample prior to analysis of the drug.

A validated stripping voltammetric procedure for quantification of the anti-hypertensive and benign prostatic hyperplasia drug terazosin in tablets and human serum.

The electrochemical behavior of terazosin at the hanging mercury drop electrode was studied in Britton-Robinson buffer (pH 2-11), acetate buffer (4.5-5.5), and in 0.1M solution of each of sodium sulfate, sodium nitrate, sodium perchlorate and potassium chloride as supporting electrolytes. The square-wave adsorptive cathodic stripping voltammogram of terazosin exhibited a single well-defined two-electron irreversible cathodic peak which may be attributed to the reduction of CO double bond of the drug molecule. A fully validated, simple, high sensitive, precise and inexpensive square-wave adsorptive cathodic stripping voltammetric procedure was described for determination of terazosin in bulk form, tablets and human serum. A mean recovery for 1x10(-8)M terazosin in bulk form, following preconcentration onto the hanging mercury drop electrode for 60s at a -1.0V (versus Ag/AgCl/KCl(s)), of 99+/-0.7% (n=5) was obtained. Limits of detection (LOD) and quantitation (LOQ) of 1.5x10(-11) and 5x10(-11)M bulk terazosin were achieved, respectively. The proposed procedure was successfully applied to determination of the drug in its Itrin((R)) tablets and human serum samples. The achieved LOD and LOQ of the drug in human serum samples were 5.3x10(-11) and 1.8x10(-10)M THD, respectively. The pharmacokinetic parameters of the drug in human plasma were estimated as: C(max)=77.5ngml(-1), t(max)=1.75h, AUC(0-t)=602.3nghml(-1), K(e)=0.088h(-1) and t(1/2)=11.32h) which are favorably compared with those reported in literature.

Voltammetric behavior and assay of the contraceptive drug levonorgestrel in bulk, tablets, and human serum at a mercury electrode.

The cyclic voltammograms of levonorgestrel (LNG) in Britton-Robinson buffers of pH 2-11 at the hanging mercury drop electrode showed a single two-electron irreversible cathodic peak over the whole pH range. This peak may be attributed to the reduction of the 3-keto-delta-4-group in the A-ring of the LNG molecule. The interfacial adsorptive character of levonorgestrel onto the surface of the hanging mercury drop electrode was identified by means of both cyclic voltammetry and chronocoulometry techniques. A simple, sensitive, and selective square-wave adsorptive cathodic stripping voltammetric procedure was developed for the quantitation of levonorgestrel. Under the optimized operational conditions, the maximum developed stripping voltammetric peak current showed a linear response with concentration of the bulk LNG substance. The achieved limits of detection (LOD) and quantitation (LOQ) were 6.7x10(-10) and 2.2x10(-9)M, or 4.8x10(-10) and 1.6x10(-9)M, following accumulation onto the hanging mercury drop electrode for 90s or 150s, respectively. The developed procedure was successfully applied to the determination of levonorgestrel in tablets, in spiked human serum, and in real plasma samples of healthy female volunteers following an oral administration of a 30-mug LNG single dose. The pharmacokinetic parameters (C(max)=1.05ngml(-1), t(max)=2.4h and AUC(0-t)=16.5nghml(-1)) were estimated and favorably compared to those reported in literature for equivalent dose.

Simultaneous determination of Cd, Pb, Cu, Sb, Bi, Se, Zn, Mn, Ni, Co and Fe in water samples by differential pulse stripping voltammetry at a hanging mercury drop electrode.

A highly sensitive and selective voltammetric procedure is described for the simultaneous determination of eleven elements (Cd, Pb, Cu, Sb, Bi, Se, Zn, Mn, Ni, Co and Fe) in water samples. Firstly, differential pulse anodic stripping voltammetry (DPASV) with a hanging mercury drop electrode (HMDE) is used for the direct simultaneous determination of Cd, Pb, Cu, Sb and Bi in 0.1 M HCI solution (pH = 1) containing 2 M NaCl. Then, differential pulse cathodic stripping voltammetry (DPCSV) is used for the determination of Se in the same solution. Zn is subsequently determined by DPASV after raising the pH of the same solution to pH 4. Next, the pH of the medium is raised to pH 8.5 by adding NH3/NH4Cl buffer solution for the determination of Mn by DPASV. Ni and Co are determined in the same solution by differential pulse adsorptive stripping voltammetry (DPAdSV) after adding DMG (1 x 10(-4) M). Finally, 1 x 10(-5) M 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol (5-Br-PADAP) is added to the solution for the determination of Fe by DPAdSV. The optimal conditions are described. Relative standard deviations and relative errors are calculated for the eleven elements at three different concentration levels. The lower detection limits for the investigated elements range from 1.11 x 10(-10) to 1.05 x 10(-9)M, depending on the element determined. The proposed analysis scheme was applied for the determination of these eleven elements in some ground water samples.