Intact Human Subjects Research Articles

Biomechanics of human quadriceps muscles during electrical stimulation

The quadriceps muscles of neurologically intact and spinal cord injured (SCI) human subjects were stimulated with constant current pulses. Up to three, separately adjustable stimulating electrodes over the motor points for vastus medialis (VM), vastus lateralis (VL) and rectus femoris (RF) muscles were used to maximize torque generation while minimizing discomfort. The torque generated by stimulation increased as the knee was slowly flexed to about 1 rad (50–60°) and decreased beyond that point (a ‘negative slope’ on a torque-angle curve). Despite this region of negative slope the force generated by small osillations remained positively correlated to the angle changes. When the knee was slowly extended again from a flexed position, the torque continued to decline and therefore showed a large degree of ‘hysteresis’. Of the three heads studied, only stimulation of RF muscle generally produced this behavior. VL and VM had torques that increased monotonically with knee flexion over the range studied. The torques generated with electrical stimulation of normal subjects represented up to about 30% of maximum voluntary contraction. When subjects generated similar torques voluntarily, the negative slope region and substantial hysteresis were not observed. Thus, SCI subjects may be adversely affected by hysteresis during electrically-induced transitions from sitting to standing and vice versa, while normal subjects are not.

Spinal Cord-Evoked Potentials and Muscle Responses Evoked by Transcranial Magnetic Stimulation in 10 Awake Human Subjects

Transcranial magnetic stimulation (TCMS) causes leg muscle contractions, but the neural structures in the brain that are activated by TCMS and their relationship to these leg muscle responses are not clearly understood. To elucidate this, we concomitantly recorded leg muscle responses and thoracic spinal cord-evoked potentials (SCEPs) after TCMS for the first time in 10 awake, neurologically intact human subjects. In this report we provide evidence of direct and indirect activation of corticospinal neurons after TCMS. In three subjects, SCEP threshold (T) stimulus intensities recruited both the D wave (direct activation of corticospinal neurons) and the first I wave (I1, indirect activation of corticospinal neurons). In one subject, the D, I1, and I2 waves were recruited simultaneously, and in another subject, the I1 and I2 waves were recruited simultaneously. In the remaining five subjects, only the I1 wave was recruited first. More waves were recruited as the stimulus intensity increased. The presence of D and I waves in all subjects at low stimulus intensities verified that TCMS directly and indirectly activated corticospinal neurons supplying the lower extremities. Leg muscle responses were usually contingent on the SCEP containing at least four waves (D, I1, I2, and I3).

Double dissociation of processing temporal and spatial information in working memory

Based on the converging evidence supporting the view of domain specific object and spatial working memory processes, the question was addressed whether the property of domain specificity holds equally for temporal information. Using a selective interference paradigm the objective was to test a dissociation of the processing of temporal duration and spatial location information in working memory of intact human subjects. Subjects performed a temporal and a spatial memory task in which they were required to indicate whether the study and the test stimuli were the same or different in duration (temporal memory) or in location (spatial memory) as primary tasks. Both primary tasks were combined with three types of interference tasks, a spatial classification memory task, a temporal classification memory task and a non-interference baseline task—to be performed in-between the presentation of study and test stimuli. Memory for temporal duration was shown to be impaired by the temporal classification task but not by the spatial classification task; memory for spatial position showed the opposite pattern of impairment. These data thus provide evidence for the view that temporal and spatial working memory contents are subject to selective interference, reflecting a functional dissociation in the processing of temporal duration and spatial location information. The results are interpreted as evidence for the domain specificity in the processing of temporal information in working memory.

Amrinone: its effect on vascular resistance and capacitance in human subjects.

Reports evaluating amrinone's effects in normal (intact) human subjects are complex and difficult to interpret because of the drug's diverse effects on myocardial function, resistance vessels, and vascular capacitance. In this study, 1.5 mg/kg of amrinone was administered as an intravenous bolus to cardiac surgical patients during constant flow cardiopulmonary bypass to determine its isolated effects on venous capacitance and vascular resistance. We noted a significant decrease in vascular resistance up to 8 min after injection and a 460 +/- 160 ml (mean +/- SD) decrease in venous reservoir volume 10 min after injection. Amrinone has potent venous and arterial vasodilating properties that make it a unique drug for treating congestive heart failure in addition to its reported positive inotropic effects.

Diminished contractile response to increased heart rate in intact human left ventricular hypertrophy. Systolic versus diastolic determinants.

Experimental studies indicate that in addition to diastolic dysfunction, hypertrophied myocardium can display depressed contractile responses, particularly at rapid heart rates, compounding reserve limitations. This study tests whether such abnormalities exist in intact human subjects at physiological paced rates and, if so, whether they are linked to simultaneous rate-dependent deterioration in diastolic function. Ten subjects with left ventricular hypertrophy (LVH) and 8 normal control subjects were studied. Most LVH patients presented with dyspnea and/or pulmonary edema and had concentric hypertrophy. Since rapid pacing simultaneously alters cardiac filling volumes and pressures, pressure-volume relation analysis was used to better define changes in contractile response. Patients were instrumented with a conductance catheter and micromanometer for pressure-volume data recording and a balloon occluder at the right atrial-inferior vena caval junction to vary filling and thus generate function relations. Data were obtained at baseline and at three atrial pacing rates (100, 120, 150 min-1). In addition, single-beat force-interval data were used to indirectly examine calcium cycling kinetics. LVH subjects demonstrated baseline diastolic abnormalities, including prolonged relaxation, elevated end-diastolic pressure, and reduced chamber compliance. However, systolic function was similar to that in control subjects. With rapid pacing, normal subjects displayed a positive contractile response, whereas this was markedly diminished in LVH subjects. With abrupt termination of pacing and return to slower sinus rhythm, LVH subjects displayed greater initial potentiation followed by a more rapid decline than control subjects, suggesting abnormalities of calcium handling. Despite contractile abnormalities, diastolic function did not further deteriorate with rapid pacing and thus did not appear to be tightly linked to the systolic changes. Pacing stress in intact human LVH can result in systolic impairment superimposed on preexisting but not worsened diastolic dysfunction. Abnormal calcium handling probably contributes prominently to this response.

Multiple descending corticospinal volleys demonstrated by changes of the wrist flexor H‐reflex to magnetic motor cortex stimulation in intact human subjects

To study noninvasively multiple descending volleys on alpha motoneurons, we used a 30-50% maximum H-reflex in the wrist flexor muscle conditioned by a subthreshold magnetic stimulus of the contralateral motor cortex as a parameter of motoneuronal excitability in seven neurologically healthy subjects. The time interval of the median nerve (test) stimulus and the magnetic (conditioning) stimulus was varied. In all subjects the conditioned H-reflex significantly (P < 0.05) enhanced at conditioning-test intervals between -1 ms and +10 ms. Superimposed on the overall facilitation there were at least two short duration periods of enhanced facilitation in all but one subject. These periods were separated by 2-4 ms and suggest increased excitability at the alpha motoneuronal pool brought on by the descending volleys. We conclude that with subthreshold magnetic cortical stimuli it is possible to produce multiple descending corticospinal volleys, which probably represent descending I waves. This should be taken into account when analyzing motor evoked potentials (MEPs) in clinical studies.

Double dissociation of spatial and object visual memory: Evidence from selective interference in intact human subjects

A functional dissociation of the spatial and object visual systems was produced by selective interference in intact young adults. Subjects were instructed to remember the location of a dot in a spatial memory test, and the form of an object in an object memory test. As predicted by current notions of dissociable visual systems in the primate, spatial memory was selectively impaired by a movement discrimination spatial task, whereas object memory was selectively impaired by a color discrimination object task.

Use of [15N]glycine in the measurement of apolipoprotein B synthesis in perfused rat liver.

Rat livers were perfused with [15N]glycine and unlabeled sodium benzoate by the single-pass technique via the portal vein or in retrograde fashion via the inferior vena cava. Perfusate [15N]hippurate enrichment was significantly greater than that of hepatic free glycine from 15 to 90 min, regardless of the direction of the perfusion. This result implies that differential labeling by periportal versus perivenous hepatocytes is not likely. When fasted animals were compared to those fed a chow diet or a sucrose-enriched diet, the labeling ratio of medium hippurate/hepatic free glycine decreased by only 9% in spite of a 5-fold decrease in the concentration of intrahepatic free glycine. Administration of nembutal to the intact animal significantly increased the enrichment of medium hippurate by 24% but did not affect the enrichment of the hepatic free glycine. We conclude that the difference between hippurate and free glycine enrichment is related to intracellular compartmentation of glycine transport. We suggest that measurement of the enrichment of hippurate after the administration of [15N]glycine with benzoate in intact animals or human subjects can therefore be used to estimate the enrichment of the intracellular precursor pool of glycine with a correction factor that does not vary appreciably under fed or fasted conditions. When uniformly labeled deuteroglycine was used as the tracer, enrichment of hepatic free glycine was decreased fivefold compared with [15N]glycine. Isotopic enrichments of apoBH and apoBL from the d less than 1.063 g/ml lipoprotein fraction isolated from the perfusion medium between 30 and 90 min averaged 3.7 and 4.1% excess, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Modulation of motor cortical excitability by electrical stimulation over the cerebellum in man.

1. We have stimulated over the cerebellum of intact human subjects by applying single electrical stimuli through electrodes placed on the back of the head, approximately at the level of the inion. The intensity of stimulation used was below that required to produce direct EMG responses in pre-activated muscles of the hand. 2. In ten subjects the effect of the stimulus over the cerebellum was to reduce the size of the EMG response in first dorsal interosseous muscle evoked by a magnetic stimulus to the cerebral cortex. In all subjects the onset of the period of suppression occurred when the test magnetic cortical shock followed the conditioning cerebellar shock by 5 ms. The duration of the suppression lasted from 3 to 7 ms. 3. The amount of suppression was related to the intensity of stimulation over the cerebellum. At 15% below the threshold for direct motor activation there was no effect; increasing suppression was evident at 10, 5 and 0% below motor threshold. 4. With a conditioning-test interval of 5-6 ms the suppression was the same whether the target muscle was relaxed or active. With longer conditioning-test intervals (12 and 15 ms) the amount of suppression was greater in active than relaxed muscles. 5. The short-latency suppression was greatest when the stimulating anode was ipsilateral to the target muscle and contralateral to the stimulated sensorimotor cortex. The later period of suppression was insensitive to the polarity of stimulation. When the stimulating electrodes were moved 2 cm caudally or cranially the short latency suppression disappeared whereas the longer latency suppression was still observed with the electrodes in the lower position. 6. Different results were obtained when the test EMG response was produced by an electrical (rather than magnetic) stimulus over the sensorimotor cortex. The short latency effect was no longer visible whereas the longer latency effect was the same as when testing with a magnetic cortical stimulus. 7. We suggest that a single electrical stimulus across the base of the skull (particularly with the anode over one cerebellar hemisphere) produces a short latency (5-6 ms) disfacilitation of the contralateral motor cortex through activation of cerebellar structures. A later (12 and 15 ms), less specific suppression which is present when testing in active muscles is thought to be mediated by a different mechanism and probably produces its effect at the level of the spinal cord.

Phantom contours: A new class of visual patterns that selectively activates the magnocellular pathway in man

The visual system of primates has two anatomical pathways: magno (M) and parvo (P). Here we report a novel technique for selectively stimulating the magnocellular pathway in man. The stimulus was a texture border between black spots and white spots displayed on a uniform gray field. This stimulus (Frame 1) was followed by Frame 2, in which all the black spots were replaced with white spots and all the white spots with black. The procedure was repeated in a continuous cycle; that is, spots reversed polarity without changing positions. At high temporal frequencies (20 Hz), subjects could not see the difference between the flickering spots, but could see a phantom contour separating the two indiscriminable regions. The difference between the spots themselves could be discriminated only at about 17 Hz. Since the M pathway can follow high flicker rates but is insensitive to the sign of the border, we suggest that the phantom border is seen exclusively by the M system. The P system, on the other hand, can report the sign of the border, but only at low flicker rates. Phantom contours provide a psychophysical scalpel for producing a temporary “lesion” in the par-vocellular pathway of intact human subjects. One could therefore repeat all of classical psychophysics and physiology using phantom contours to determine whether or not a given visual process receives an M input. Also, these stimuli might provide a simple diagnostic test for revealing the loss of M-cell function that occurs in early glaucoma.

Positive Inotropic Effects Mediated by α1 Adrenoceptors in Intact Human Subjects

The role of alpha 1-adrenergic receptors (adrenoceptors) on cardiac contractility was investigated in human subjects. The effect of methoxamine, a selective alpha-adrenoceptor agonist, and angiotensin II, on cardiac contractility was determined by means of noninvasive assessment of the slope of the end-systolic pressure (ESP)/end-systolic dimension (ESD) relationship. The slope (m) of this ratio was significantly higher with methoxamine (17.0; SD = 9.0 mm Hg/mm) than with angiotensin II (4.8; SD = 1.9 mm Hg/mm) (p less than 0.05). Slopes with methoxamine were higher when heart rates (HRs) were reflexly reduced, and were significantly diminished when reflex bradycardia was prevented by atropine (p less than 0.05) or atrial pacing (p less than 0.01). Previous treatment with propranolol did not modify m values with methoxamine (m = 15.5; SD = 4.4 mm Hg/mm). Phentolamine, given at peak methoxamine effect, did not consistently modify m values, resulting in an average slope not significantly different from that obtained with methoxamine alone. However, the addition of phentolamine did not cause an increase in ESDs at each level of ESP with respect to methoxamine. In the same subjects, infusion of phentolamine after angiotensin did not modify ESDs at comparable ESP levels. These findings suggest the existence of a positive inotropic effect mediated by alpha 1 adrenoceptors in the intact human heart.

Functional organization of the trigeminal motor system in man. A neurophysiological study.

Transcranial stimulation (TCS) in intact human subjects was used to investigate the corticobulbar projections and the functional organization of the trigeminal motor system. Both electrical (with the anode overlying the face area of the motor cortex) and magnetic TCS (with the coil at the vertex) excite the upper motoneurons projecting to the trigeminal motor nucleus, evoking motor potentials (C-MEPs) in the jaw-closing and suprahyoid muscles, but only during voluntary contraction. At least 30% of jaw-closing motoneurons are reached by direct fast-conducting corticobulbar fibres; these projections are mainly crossed. Suprahyoid motoneurons are also reached by fast-conducting corticobulbar fibres; these projections are probably bilateral. In the masseter, electrical TCS also evokes an ipsilateral motor response (R-MEP), followed by a later wave (U), and bilateral inhibitory periods. The R-MEP is secondary to excitation of the motor trigeminal root; the U wave probably results from the simultaneous excitation of Ia afferents in the root and ipsilaterally projecting corticofugal fibres; the inhibitory periods are largely due to activation of exteroceptive afferents in the root. Magnetic TCS, avoiding spread of current to the trigeminal root, evokes C-MEPs but not R-MEPs or U waves. The masseter inhibitory period after magnetic TCS may be due to excitation of corticofugal inhibitory fibres and to mechanical activation of Golgi tendon organs.

Electromyographic Studies of Motor Control in Humans

Electromyography and electroneurography have proved to be useful in investigation and understanding of a variety of neurologic disorders. In most laboratories, however, these electrodiagnostic techniques have been used to help in the diagnosis of diseases that affect the peripheral nerves, neuromuscular junctions, or skeletal muscle fibers. Although major advances in electronic and computer technology have made it possible to study, quantitate, and document reflex activity in intact human subjects, most neurologists still rely on gross clinical observations and most electromyographers continue to use conventional techniques of EMG and nerve conduction studies to differentiate "myopathy" from "neuropathy." This article is a review of some of the electromyographic techniques that have been used in the authors' laboratory for the study of normal and abnormal motor control in man and the treatment of patients with disorders of motor control.

Characterization and control of muscle response to electrical stimulation.

The maintenance of upright posture in neurologically intact human subjects is mediated by two major nervous pathways. The first, leading from the cerebral cortex through the spinal cord to motor neurons, activates muscles which produce postural movements. The second, leading from various sensory organs to higher centers, provides sensory feedback regarding the postural state. The path through the spinal cord is no longer intact in victims of spinal cord injury and loss of normal control of muscle activity results. Functional neuromuscular stimulation (FNS) has been shown as a feasible method for obtaining muscle contraction in paraplegics and has been proposed as a means for control of antero-posterior sway to make upright posture possible for these individuals. Before muscle can be controlled through the use of FNS, the response of muscle to electrical stimulation must be understood. In past studies, linear control theory has been applied to the analysis of this response and to the testing of various controllers. The aim of this study was to demonstrate some control issues in FNS using linear control theory, as it applies to electrical stimulation of muscle for stabilization of posture. The linearity of the muscle response was improved through closed-loop control using pole compensation techniques. The excess phase shift of the system due to the time delay in the muscle response, however, limits the ability to increase the open-loop gain in order to obtain improved performance. A suggestion for further study is the application of this methodology for uses in posture control.

Number and affinity of insulin receptors in intact human subjects

A simple model of the distribution and metabolism of insulin in vivo has been evaluated using data from insulin infusion into a group of normal subjects. The major rate-limiting step for access to degradation pathways is assumed to consist of binding of the ligand to a single population of insulin receptor sites, except that provision is made for the possibility of linear non-receptor-mediated degradation and for the phenomenon of negative cooperativity. The model has been shown to accommodate the non-linearity of insulin metabolism, allows, evaluation of receptor association and dissociation constants and provides for the first time an estimate of total accessible receptor number in the intact organism. For normal fasting man the model predicts 1.00 +/- 0.05 nmol accessible binding sites/kg (mean +/- SD).

Sinoatrial function after cardiac transplantation

The function of both the denervated donor and innervated recipient sinus nodes of 14 asymptomatic cardiac transplant recipients was assessed. Tests of sinoatrial function were performed in 14 donor and 10 recipient atria. The mean spontaneous cycle length of the recipient atria was significantly longer than that of the donor atria (944 +/- 246 versus 663 +/- 158 ms, p less than 0.01). Donor sinus node recovery time was prolonged in four patients (greater than 2,500 ms in two) and recipient recovery time was prolonged in six patients. In those patients with normal sinus node function tests, the recovery time of the recipient sinus node was longer than that of the donor sinus node (1,170 +/- 207 versus 864 +/- 175 ms, p less than 0.02). The pattern of response of recovery times to increasing pacing rate was predictable and organized in the donor but chaotic in the recipient, and the longest sinus node recovery time occurred at the shortest pacing cycle length used in 12 of the 14 donor atria but in only 1 of the 10 recipient atria (p less than 0.001). Secondary pauses occurred in none of the normal donor atria and in all of the abnormal donor atria (p less than 0.001); however, they occurred in both normal and abnormal recipient atria. The recipient and donor atria were paced alone and synchronously in the same patients. Synchronous pacing had no effect on the recovery times of the donor sinus node but significantly lengthened those of the recipient (sinus node recovery time: 1,266 +/- 218 to 1,547 +/- 332 ms, p less than 0.02; corrected recovery time: 322 +/- 102 to 686 +/- 188 ms, p less than 0.01). In the donor atria, abnormal recovery time was invariably associated with abnormal sinoatrial conduction time. There was a strong correlation between sinoatrial conduction time measured by the methods of Strauss and Narula and their coworkers in the donor atria (r = 0.98, p less than 0.001) but not in the recipient atria (r = 0.72). In the absence of autonomic influences, tests of sinus node function of the donor atria produce predictable and consistent results and, therefore, may be more clinically reliable than in intact human subjects. There is a high incidence of recipient sinus node dysfunction in asymptomatic long-term survivors of cardiac transplantation.

Effects of motor cortex stimulation on spinal interneurones in intact man.

The effect of a descending corticospinal volley on a spinal inhibitory pathway, has been studied in five intact human subjects. Approximately 63% inhibition of the H-reflex evoked in wrist and finger flexor muscles, was produced by motor threshold stimulation of the radial nerve. When a submotor threshold cortical shock was given 2 to 4 ms before the H-reflex, this inhibition was reduced to approximately 38%. The timing of this effect is compatible with either a monosynaptic or disynaptic corticospinal tract projection onto the spinal inhibitory interneurone.

Accuracy of volume determination by two-dimensional echocardiography: defining requirements under controlled conditions in the ejecting canine left ventricle.

The accuracy of two-dimensional echocardiographic left ventricular volume measurement in an isolated heart preparation was tested using Simpson's reconstruction of progressively fewer short-axis cross sections of known location. Echocardiographic images from five ejecting hearts submerged in a special tank were obtained under conditions designed for maximal accuracy of echocardiographic volume assessment. Echocardiographic determinations of 52 volumes at various times throughout the cardiac cycle were compared, by least-squares linear regression, with simultaneous direct-volume measurements by volumetric chamber (range 9.4-44.8 ml). Echocardiographic and direct measurements correlated well for all numbers of cross sections from 1-19 (r = 0.84-0.97); however, variability of direct volume predicted from a given echocardiographic measurement increased nonlinearly as the number of cross sections per heart decreased, and was especially large when three or fewer cross sections were used (SEE = 4.6-7 ml). The accuracy of echocardiographic measures was compared for each number of cross sections per heart, varying from one to 19; accuracy was defined as the mean absolute difference between echocardiographic and direct measurements of volume, ejection fraction, and maximal rate of ejection. The accuracy of echocardiographic measurements was significantly reduced with fewer than four cross sections per heart for ventricular volume, three cross sections for ejection fraction, and five cross sections for maximal rate of ejection. In light of what appears to be required for accurate echocardiographic volume measurement in this controlled, ejecting, noninfarcted, in vitro preparation, additional cross sections may be required in intact animals and human subjects, especially in those with diseases that cause ventricular asymmetry or regional dysfunction.

Premature atrial stimulation as a key to the understanding of sinoatrial conduction in man. Presentation of data and critical review of the literature.

Since recording potentials directly from the sinoatrial node (SAN) is not yet possible, the electrophysiologic evaluation of this structure in intact human subjects must be accomplished with indirect technics. Two technics have been used to study SAN function in man: premature atrial stimulation (PAS) and rapid atrial pacing. Recent discussions of data using these technics have emphasized their role in determining SAN automaticity, but their role in evaluating conduction from atrium to SAN or SAN to atrium has not been fully explored. Using the technic of PAS, we have studied five patients with sinus bradycardia and symptoms of dizziness or syncope. Our analysis of the results obtained from these studies discloses the unique ability of this technic to evaluate conduction into and out of the SAN. An atrial premature depolarization (APD) elicited late in atrial diastole is followed by a compensatory pause (nonreset of the SAN pacemaker). An APD elicited earlier in atrial diastole is followed by a pause that is less than compensatory (SAN reset). From these responses estimates of sinoatrial conduction time were made. In one patient reset was never seen, suggesting markedly prolonged sinoatrial conduction. With these results in mind, the literature was reviewed and an alternate interpretation posed for existing data. PAS is not only a means of determining SAN automaticity, but also a very useful means of unmasking sinoatrial conduction abnormalities.

Direct study of left atrial P waves

Direct electrical stimulation of the left atrium in intact human subjects was performed in 11 patients. The resultant P waves did not conform to the strict left to right direction suggested by previously published studies. Marked variability was observed in the P waves resulting from electrical stimulation of the same area of the left atrium in different subjects. The existing criteria for diagnosis of left atrial P waves based on preconceived notions could not be corroborated. No new criteria could be established, and none is proposed. Recording of an esophageal or intracavitary atrial lead simultaneously with surface leads should be regarded as the first, indispensable prerequisite in any future study of left atrial P waves. It is suggested that stimulation of a given point on the walls of the left atrium may give rise to varying P waves probably because of the interplay of slow, wave-like intramyocardial conduction with the more rapid, non-wave-like spread via the specialized interatrial pathways. P waves resulting from electrical stimulation of the right lower aspect of the interatrial septum at or near the coronary sinus were virtually indistinguishable from left atrial P waves.