Analysis of the causes of increased tremor in thyroid hyperfuction]

The authors present the results of examination of tremor (hand and trunk) of the amplitude of H- and T-reflexes and of the H-reflex inhibition during the contraction of antagonistic muscles and of facilitattion of T- and H-reflexes during the Yendrassik maneuvre in 33 children with hyperfunction of the thyroid gland and 20 normal subjects. It was established that the amplitude of tremor was greater and the frequency was less in these patients than in normal subjects; the amplitude of the H- and T-reflexes was enhanced, and the intensity of reciprocal inhibition of the soleus muscle motor neurons was less in the patients. Comparison of the amplitude of the T- and H-reflexes indicated that against the background of the Yendrassik maneuvre facilitation of the T-reflex was less intense and facilitation of the H-reflex was more intense than in normal subjects. The authors supposed that the enhanced tremor in the patients was due to the motor neurons pool depolarization, to decrease the efficacy of reciprocal inhibition and to diminished suprasegmental influences spread to the gamma-motor neurons.     

Mechanism of hypoprothrombinemia and hypoproconvertinemia in experimental thyrotoxicosis]

The changes in the thromboelastogram (TEG), the activity and disintegration of prothrombin and proconvertin were studied in experiments on rats with thyrotoxicosis induced by giving thyroidin per os. In marked thyroidin intoxication the majority of TEG indices shifted to hypocoagulation. The most distinct changes were: delayed thrombin formation, an increase in the general coagulation constant, and a decrease in the maximal amplitude. Besides, a decrease of the activity and an increase in prothrombin disintegration (particularly of proconvertin) was observed. It is supposed that increased disintegration of these procoagulants is of great importance in the pathogenesis of hypoprothrombinemia and hypoproconvertinemia in thyrotoxicosis.     

Chronic myocardial infarction induces phenotypic and functional remodeling in the guinea pig cardiac plexus

Chronic myocardial infarction (CMI) is associated with remodeling of the ventricle and evokes adaption in the cardiac neurohumoral control systems. To evaluate the remodeling of the intrinsic cardiac nervous system following myocardial infarction, the dorsal descending coronary artery was ligated in the guinea pig heart and the animals were allowed to recover for 7-9 wk. Thereafter, atrial neurons of the intrinsic cardiac plexus were isolated for electrophysiological and immunohistochemical analyses. Intracellular voltage recordings from intrinsic cardiac neurons demonstrated no significant changes in passive membrane properties or action potential configuration compared with age-matched controls and sham-operated animals. The intrinsic cardiac neurons from chronic infarcted hearts did demonstrate an increase in evoked action potential (AP) frequency (as determined by the number of APs produced with depolarizing stimuli) and an increase in responses to exogenously applied histamine compared with sham and age-matched controls. Conversely, pituitary adenylate cyclase-activating polypeptide (PACAP)-induced increases in intrinsic cardiac neuron-evoked AP frequency were similar between control and CMI animals. Immunohistochemical analysis demonstrated a threefold increase in percentage of neurons immunoreactive for neuronal nitric oxide synthase (NOS) in CMI animals compared with control and the additional expression of inducible NOS by some neurons, which was not evident in control animals. Finally, the density of mast cells within the intrinsic cardiac plexus was increased threefold in preparations from CMI animals. These results indicate that CMI induces a differential remodeling of intrinsic cardiac neurons and functional upregulation of neuronal responsiveness to specific neuromodulators.     

Direct and indirect assessment of gamma-motor firing patterns

The study of the patterns of gamma-motor activity which accompany natural contractions has been long and difficult, and has not as yet led to general agreement. In this review we have simplified matters by considering the case of locomotion in the cat only, and we have avoided discussion of the various hypotheses which have been advanced to provide general schemes of gamma control for a wide range of movements. The development of the subject is shown to depend very much on devising ingenious methods applicable to reduced and intact animals. Direct recording from gamma-motoneurones has only been possible in reduced preparations, whereas indirect assessment of gamma activity from spindle afferent recordings was used in these and in intact animals. At this point in time, we still have no direct recordings from gamma-motoneurones in normally behaving animals, but those obtained in decerebrate animals show distinct patterns of modulation for static and dynamic types with particular temporal relation to the stepping movements. The spindle recordings in intact animals potentially provide the most important information, and the problems of interpretation, which have previously caused difficulties, are beginning to be solved through the insights obtained from the reduced preparations.     

Restriction of stress-induced activation of lipid peroxidation by small doses of thyroid hormones]

The experiments on 57 female rats demonstrated that small doses of thyroid hormones (thyroidin) significantly (55-118%) restrict stress induced increase in the concentration of initial and terminal products of lipid peroxidation (LP) in the myocardium and in the blood plasma. After hormone injection stress decreases the activity of key antioxidant enzyme, superoxide dismutase of erythrocytes (SOD), to a lesser degree and increases the rate of malonyldialdehyde (MDA) production induced by Fe2+ in homogenates of the myocardium to the same degree as well in comparison with rats that had not been injected thyroidin. In normal rates thyroidin does not influence the concentration of products of LP, increases the activity of SOD and decreases increment of MDA induced by Fe2+ in homogenates of the myocardium. Thus, small doses of thyroid hormones restrict significantly stress induced activity of LP membranes, increasing the power of antioxidant systems both in the myocardium and in the organism.     

Functional Reinnervation of the Distal Hindlimb Muscles in Rats after Compression of the Sciatic Nerve

Chronic experiments (12 weeks long) were carried out on two groups of pubertate-age Wistar rats. The sciatic nerve in one hindlimb was compressed, and complex analysis of the dynamics of functional reinnervation of the distal hindlimb muscles was performed. We used videorecording of images of the feet in the course of locomotion for estimation of the level of functional reinnervation of the toe extensors (the technique was modified in our laboratory) and a tensometric technique (measuring of the contraction force of the toe flexors and ankle extensors in the course of realization of the burrowing instinct by the animals). In the first group of animals (n = 21), we studied the dynamics of recovery of the force developed by the toe flexors and ankle extensors and of the functional sciatic index after sciatic nerve traumatization with no additional pharmacological influences. In the second group (n = 29), the same indices were analyzed, but after single injections of gamma-hydroxybutyrate (GHB) into animals before compression of the sciatic nerve and within early and late (3–4 and 14 weeks, respectively) terms after such intervention. Within the first week after compression, the smallest loss of activity was observed in the toe flexors, as compared with that in the toe and ankle extensors. Significant functional reinnervation of the ankle and toe extensors (to 52 and 87%, respectively) was observed on the 3rd to 7th weeks after the nerve injury. Functional reinnervation of the toe flexors was characterized by a relatively greater initial force of concentrations and more rapid recovery than those of the ankle and toe extensors. Practically complete recovery of the functions of the toe flexors was observed on the 10th week, while that of the ankle extensors was found on the 12th week. To make clearer possible reasons for differences between the processes of recovery of functions of the flexor and extensor muscles, we analyzed the effects of GHB on the contractile activity of these muscle groups within different time intervals after traumatization. In the second animal group, injection of GHB before operation resulted in drops of the muscle contraction force. After injury of the right sciatic nerve and injection of GHB on the 3rd posttraumatic week, recovery of the contraction force of all the examined muscles on the side of operation was more considerable. The functional loss of the muscle force in the left (intact) hindlimb in this case was 10 and 7% for the ankle extensors and toe flexors, respectively. Single injections of GHB within a late posttraumatic period resulted in a relative decrease in the contraction force of both flexors and extensors, and the functional loss was the greatest (35%) in the toe flexors.     

Parameters of discharges of respiratory neurons of the ventrolateral medullary regions in early postnatal rats

In experiments on superfusedin situ semi-isolated medullo-spinal preparations (SIMSP) of newborn (1st day of life) and 4- to 5-day-old rats, we studied the parameters of extracellularly recorded spike activity of respiratory neurons of the ventrolateral medullary regions (VLMR). In SIMSP of 4- to 5-day-old rats, the frequency of discharges of pre-inspiratory, inspiratory, and expiratory neurons is shown to be significantly higher, while the dispersion of its values is considerably lower, as compared with the corresponding values for newborn animals. In the majority of pre-inspiratory and inspiratory neurons of SIMSP of newborn rats, irregular low-frequency discharges are usually generated within the interinspiration phase. The relative intensity of suppression of discharges of pre-inspiratory and expiratory neurons within an inspiration phase is much lower in SIMSP of newborn rats, as compared with that in 4- to 5-day-old preparations. The activity of most pre-inspiratory neurons manifests a trend toward transformation from a two-phase pattern in newborn rats (with two frequency peaks, pre- and post-inspiratory) to a monophasic pattern (with one pre-inspiratory frequency peak) typical of 4- to 5-day-old animals. The effects of electrical stimulation of the site of localization of pre-inspiratory neurons showed that in SIMSP of both age groups of rats an inspiratory response could be evoked in then. phrenicus only in the case when stimulation was applied within the second half of an interinspiratory phase. Therefore, it can be supposed that the respiratory network in newborn animals is to a considerable extent immature in the morphofunctional aspect. It seems probable that in early postnatal rats pre-inspiratory neurons are involved in the medullary mechanisms foron-off switching of the inspiratory and expiratory phases.Neirofiziologiya/Neurophysiology, Vol. 28, No. 4/5, pp. 207–217, July–October, 1996.     

The somatotopy of the masticatory neurons in the rat trigeminal motor nucleus as revealed by HRP study

Young adult albino rats of Wistar strain were used for the present study. 0.5 to 15 microliters of 20-50% of horseradish peroxidase (HRP) were injected into each individual muscle of mastication to label neurons in the trigeminal motor nucleus (TMON) for light microscopic study. The results reveal that: (1) Many HRP-labeled, multipolar neurons are observed in the motor nucleus in each jaw-closing muscle (JCM) with less in each the jaw-opening muscle (JOM). (2) The motor neurons innervating each masticatory muscle in the motor nucleus show a somatotopic arrangement: (a) those innervating the temporalis muscle are located in the medial and dorsomedial parts; (b) those innervating the masseter muscle are located in the intermediate and lateral; (c) those innervating the medial and lateral pterygoid muscles are located in the lateral, ventrolateral and ventromedial parts, respectively; and (d) those innervating the mylohyoid and the anterior belly of the digastric muscles are located in the most ventromedial part of the caudal one-third of the nucleus. Axons of most masticatory motor neurons run ventrolaterally in between the motor and the chief sensory nuclei of the trigeminal nerve. However, those of the mylohyoid and anterior belly of the digastric muscles ascend dorsally to the dorsal aspect of the caudal nucleus and then turn ventrolaterally to join the motor root of the trigeminal nerve. Furthermore, the dendrites of the motor neuron of JCM converge dorsocaudally to the supratrigeminal region. The diameters of neurons of each JCM display a bimodal distribution. However, an unimodal distribution is present in the motor neurons from each JCM. It is suggested that the motor nucleus innervating the JCM is comprised of comprised of alpha- and gamma-motor neurons. It, thus, may provide a neural basis for the regulation of the muscle tone and biting force.     

The background and evoked neuronal impulse activity of embryonic neurotransplant of somatosensory brain cortex in rats

The investigation was intended for studying the character of the background and evoked impulse activity of embryonic neurotransplant neurons 4 months after homotopical allotransplantation into the barrel field of somatosensory recipient's brain cortex of the rat. It is established, that the current average frequency of background impulse activity of transplant neurons is reduced in comparison with one of the control rats. It is shown that the evoked impulse activity of neurotransplant develops with the long latency than in somatosensory cortex of the control animals. Thus in patterns of the evoked activity of neurotransplant cells reactions, characteristic for the neurons of barrel field somatosensory recipient's cortex of control rats are registered: an increase of frequency of pulses' generating, or alternating of the activation and reduction periods of impulse frequency with its subsequent regeneration up to a pristine level.     

弱背景噪声对大鼠听皮层神经元频率调谐的影响

在自然环境中,人和动物常在一定的背景噪声下感知信号声刺激,然而,关于低强度的弱背景噪声如何影响听皮层神经元对声刺激频率的编码尚不清楚.本研究以大鼠听皮层神经元的频率反应域为研究对象,测定了阈下背景噪声对79个神经元频率反应域的影响.结果表明,弱背景噪声对大鼠初级听皮层神经元的听反应既有抑制性影响、又有易化性影响.一般来说,抑制性影响使神经元的频率调谐范围和最佳频率反应域缩小,易化性影响使神经元的频率调谐范围和最佳频率反应域增大.对于少数神经元,弱背景噪声并未显著改变其频率调谐范围,但却改变了其最佳频率反应域范围.弱背景噪声对63.64%神经元的特征频率和55.84%神经元的最低阈值无显著影响.神经元频率调谐曲线的尖部比中部更容易受到弱背景噪声的影响.该研究结果有助于我们进一步理解复杂声环境下大脑听皮层对听觉信息的编码机制.     

Physiological modifications of local haemodynamic conditions during bilateral isometric contractions

Nine subjects (five women and four men) simultaneously performed two isometric contractions sustained until exhaustion at different relative forces: 40% of maximum voluntary contraction (MVC) for the right elbow flexors; 50% MVC for the left elbow flexors. Contraction of the left elbow flexors commenced at 50% of the limit time (maximum maintenance time) of isometric contraction of the right elbow flexors. Increase in heart rate during concomitant contraction of the left elbow flexors led to an increase in blood flow to the right elbow flexors. Under these conditions, the limit time of isometric contraction of the right elbow flexors was prolonged with respect to the limit time obtained for an isolated isometric contraction at the same relative tension. The difference was more significant in the female (+40%, P less than 0.05) than in the male subjects (+20%, P greater than 0.05).     

Neuronal background of activation of estivated snails,with special attention to the monoaminergic system: a biochemical,physiological, and neuroanatomical study

Osmotic stimulation activates both estivated and inactivated specimens of Helix pomatia and increases their central arousal. High-pressure liquid chromatography has shown that, during activation, the level of both serotonin and dopamine decreases in the central nervous system (CNS) but increases in the foot and heart, organs that are involved in the eversion of the body. In isolated CNS from activated animals, the firing frequency of the heart-modulator serotonergic (RPas) neurons is significantly higher than that in the CNS of estivated or inactivated animals. These neurons innervate both the heart and the anterior aorta. In semi-intact preparations, distilled water (an osmotic stimulus) applied to the mantle collar increases their firing frequency, whereas tactile stimulation evokes their inhibition. Extracellularly applied monoamines mimic the effect of peripheral stimuli: serotonin (0.1–10 μM) increases the activity of the RPas neurons, whereas dopamine (0.1–10 μM) inhibits their activity. Tyrosine-hydroxylase immunocytochemistry and retrograde neurobiotin tracing have revealed similar bipolar receptor cells in the mantle collar and tail, organs that are exposed to environmental stimuli in estivated animals. Serotonin immunocytochemistry carried out on the same tissues does not visualize receptor cells but labels a dense network of fibers that appear to innervate neurobiotin-labeled receptor cells. The combination of neurobiotin-labeling of RPas neurons and immunolabeling suggests that RPas neurons receive direct dopaminergic inputs from receptor cells and serotonergic inputs from central serotonergic neurons, indicating that central serotonergic neurons are interconnected. Thus, the RPas neurons may belong to neuronal elements of the arousal system. This work was supported by Hungarian OTKA grants T037389, T046580, T037505, and K63451.     

Partial urethral obstruction enhances NADPH-diaphorase activity in the monkey (Macaca fascicularis) bladder: light and electron microscopic studies

The effect of partially obstructing the urethra on the nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) activity in neurons of the intramural ganglia of the monkey (Macaca fascicularis) bladder was examined by light and electron microscopy. Partial urethral ligation was done in adult male monkeys. The animals were sacrificed 2, 4 weeks after partial urethral obstruction. This was compared to controls (normal and sham operated). Urethral obstructed animals were observed to have increased urinary frequency and decreased urinary flow rate. Two weeks after urethral obstruction, the overall NADPH-d activity in the intramural ganglia of the bladder base was enhanced compared to control animals. The frequency of intensely stained NADPH-d positive neurons was increased compared to the control animals. About one-third of intensely stained NADPH-d positive neurons appeared to undergo degenerative changes. At 4 weeks after urethral obstruction, a wide occurrence of NADPH-d positive neurons in advanced stages of degeneration in the bladder base was observed. Cellular debris was strewn among normal looking ganglion cells and along the nerve processes. The proportion of intensely stained NADPH-d positive neurons was relatively lower than the controls. The total number of NADPH-d positive neurons and the nerve fibres in the entire bladder was significantly reduced when compared to control animals. Electron microscopy showed some NADPH-d activity in intramural ganglion cells in 2 weeks after partial urethral obstruction. NADPH-d reaction product (formazan) was deposited on the membranes of the rough endoplasmic reticulum, and the outer membranes of some mitochondria in the intramural neuron. At 4 weeks after urethral obstruction, NADPH-d was present in the membrane of the mitochondria and some mitochondria appeared swollen with disrupted cristae. Present results show that NADPH-d activity in neurons of the intramural ganglia of the monkey (Macaca fascicularis) urinary bladder was increased after two weeks and reduced after 4 weeks of partial urethral obstruction. It is speculated that the increased NADPH-d activity associated with partial urethral obstruction would lead to neuronal damage and death, which may contribute to detrusor overactivity. However, it warrants further investigation to understand the mechanism of neuronal cell death after partial urethral obstruction.     

Cosinor analysis of the seasonal changes in the cytoplasmic RNA level in the neurons of preoptic nucleus in Rana temporaria L. under the ultrasound energy

Authors studied the effects of single, total doses of ultrasound energy (frequency: 24.6 kHz; intensity: 0.35 W/cm2) on the cytoplasmic RNA level in the neurons of preoptic nucleus (PON) in Rana temporaria L., in the annual cycle. The cosinor analysis revealed that the applied ultrasound energy caused significant changes in the cytoplasmic RNA content in the PON neurons in Rana temporaria L. in the annual cycle. This was expressed by an increase in rhythm amplitude, except for the females of the second experimental group. Simultaneously, the rhythm acrophase in the animals exposed to ultrasound energy was shifted to later months (i.e. to the hibernation period), as compared to the control animals, where the acrophase occurred in August in females and in September in males (i.e. in their active land life period). In the females of the first experimental group the ultrasounds induced a disappearance of rhythmic changes in the cytoplasmic RNA of the PON neurons in the annual cycle.     

Late long-lasting discharges in hind limb motor nerves and their connection with the locomotor rhythm in thalamic immobilized cats

Effects of stimulation of flexor reflex afferents were studied in decerebrate immobilized cats. Stimulation of ipsilateral afferents evoked late long-lasting discharges in the nerves to the flexors, whereas stimulation of contralateral afferents led to similar discharges in nerves to both extensors and flexors. Compared with spinal animals, early segmental reflexes in thalamic cats were tonically depressed. Similar tonic inhibition of segmental reflexes took place in spinal animals after injection of dopa. Segmental reflexes were clearly modulated during late or rhythmic discharges. The possible central mechanisms of these changes in the segmental reflexes are discussed on the basis of data in the literature.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 11, No. 2, pp. 137–145, March–April, 1979.     

Activity of neurons of the cerebral A5 zone of rat induced by adequate stimulation of muscle afferents: On the control of arterial pressure and respiration during muscle activity

In experiments on 24 anesthetized rats with preserved spontaneous respiration, we first recorded the volley impulse activity of neurons (n = 30) in the brainstem A5 zone, which was induced by periodical stretchings of the forelimb flexors and hindlimb extensors. The frequency of action potentials in such volleys was, on average, 99.7 ± 19.6 sec⁻¹. In the course of this kinesthetic stimulation, along with the activation of “proprioceptive” neurons of the A5 zone, we observed transitory drops in the arterial pressure and increases first recorded the activity of baroceptive neurons in subpial parts of the A5 zone (n = 4); the frequency of their background impulsation was, on average, 25.1 ± 0.8 sec⁻¹. This activity in all cases was transitorily suppressed both upon increases of the blood pressure caused by constriction of the carotid arteries or nociceptive tail stimulation, as well as upon stretching of skeletal muscles. Therefore, we first obtained direct proof that neuronal systems of the A5 zone are involved in integration of visceral and somatic proprioceptive afferent influences. We hypothesize that the physiological role of this mechanism of integration of somatic and visceral information at the level of the A5 zone is directed toward lowering of the arterial pressure and intensification of respiration within the period of intensified motor activity. This mechanism is based on the interaction between “proprioceptive,” baroceptive, and, perhaps, multiceptive neurons within the A5 zone. Neirofiziologiya/Neurophysiology, Vol. 39, No. 6, pp. 443–452, November–December, 2007.     

Neuronal activity within the ventrolateral periaqueductal gray during simulated hemorrhage in conscious rabbits

The ventrolateral (vl) periaqueductal gray (PAG) has been proposed as a site responsible for the active process triggering the onset of hypotension (i.e., phase 2) during blood loss in conscious animals (Cavun S and Millington WR. Am J Physiol Regul Integr Comp Physiol 281: R747-R752, 2001). We recorded the extracellular activity of PAG neurons in conscious rabbits to test the hypothesis that vlPAG neurons change their firing frequency before the onset of hypotension during simulated hemorrhage. Arterial and venous catheters, an intrathoracic vena caval occluder, and midbrain microelectrodes on a microdrive were implanted in 10 rabbits. During simulated hemorrhage, the occluder was inflated until arterial pressure < or = 40 mmHg. We compared changes in neuronal activity during simulated hemorrhage with those during a similar length control period for 64 vlPAG and 29 dorsolateral (dl) PAG neurons. Arterial pressure pulse modulation of neuronal activity was present in 45 and 76% of vlPAG and dlPAG neurons, respectively. When we evaluated the absolute change in activity, thus accounting for both increases and decreases, simulated hemorrhage had a significant effect on activity of vlPAG but not dlPAG neurons. The majority (56%) of vlPAG neurons did not appear to respond to simulated hemorrhage. Of the 28 responsive vlPAG neurons, 11 showed an abrupt change in firing frequency during the time interval preceding the onset of hypotension; 13 responded after the onset of hypotension; and 4 showed a consistent direction of change across the entire simulated hemorrhage. Thus 24 (38%) of the vlPAG neurons recorded responded at a time consistent with a contribution to the hypotension associated with simulated hemorrhage.     

Age-specific morphological-functional characteristics of the rabbit's hippocampal neurons during conditioning

Formation of trace rhythm recruitment (an analogue of conditioned time reflex) was studies in CA3 hippocampal neurons of alert young (less than one year), old (54-65 months), and very old rabbits after a prolonged (10-20 min) electro-cutaneous stimulation of a forelimb with the frequency of 0.5-1 Hz. Comparative analysis of neuronal spike activity in young and old rabbits showed that in the late ontogeny the number of spontaneously active neurons was significantly decreased, the proportion of slowly firing neurons increased, the interspike intervals and intervals between spike groups became longer, the number of spikes in a group reduced. The ability of hippocampal neurons to acquire and reproduce the rhythm of the previous stimulation declined with age. No appropriate rhythms were found in neurons of very old animals. A nonspecific increase in neuronal baseline activity was observed in old rabbits after the stimulation. Deterioration of morphological structures of hippocampal neurons and glial cells may explain the impairment of mnestic processes in late ontogeny.     

Characteristics of electrical responses by cochlear nuclei in Vespertilionidae and Rhinolophidae to ultrasonic stimuli with various fill frequencies

The total electrical response and action potentials of separate neurons in the cochlear nuclei in Vespertilionidae and Rhinolophidae were investigated. Maximum sensitivity to ultrasound was recorded in Vespertilionidae in the frequency ranges 10–30 and 70–80 kc/sec, and in Rhinolophidae in the frequency ranges 10–30 and 84–86 kc/sec. Mininum off-response thresholds were observed in Vespertilionidae in the range 50–60 kc/sec, and in Rhinolophidae in the range 78–80 kc/sec. The areas of responses by neurons in the cochlear nuclei in both species of bats were similar in shape to those recorded in the same structure in other animals. An exception was provided by Rhinolophidae, in which three peculiar types of neurons were observed: 1) neurons whose response area lay in the frequency ranges up to 78 kc/sec or from 80 to 90 kc/sec; 2) neurons responding in the range 40–90 kc/sec, but not sensitive to stimuli with a fill frequency of 78–80 kc/sec; and 3) neurons whose response area lay in the range 78–80 kc/sec, but in which the character of the response changed from tonic to phasic when there was a change in the fill frequency of the stimulus. Maximum selectivity with regard to fill frequency of stimulus was observed in the neurons of Rhinolophidae in the frequency range 70–90 kc/sec.The term "fill frequency" can be rendered as frequency — Consultants Bureau.A. A. Zhdanov Leningrad State University. Translated from Neirofiziologiya, Vol. 3, No. 4, pp. 379–385, July–August, 1971.