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Effects of hypoxia and other injurious stimuli on collagen secretion and intracellular growth stimulating activity of bovine aortic smooth muscle cells in culture.

Bovine arterial smooth muscle cells in culture were incubated in hypoxia and total cellular DNA and collagen secretion during and after the hypoxic period was measured as well as the effect of conditioned medium from hypoxic cells on these parameters. Collagen secretion decreased by 16.4% compared to controls during hypoxia but was increased by 41.4% in the post-hypoxic period. Total cellular DNA was significantly lower after both periods. New cultures, receiving conditioned medium from hypoxic cultures, showed an increased collagen secretion by 32.2% compared to controls while total cellular DNA was not changed. Growth stimulating activity, previously shown to be released from lysed cultured SMC, was assayed by exposing SMC cultures to supernatant from lysed cells that had been incubated in hypoxia or exposed to other potential atherogenic stimuli. The growth stimulating activity per cell could be increased by incubating cells in hypoxia, or exposing them to low density lipoproteins or cigarette smoke condensate in concentrations high enough to cause a decrease in cell number. It was suggested that the described effects might contribute to increased cell proliferation and collagen formation in the development of atherosclerosis.

Animals↗

Effects of cortical stimulation on auditory-responsive thalamic neurones in anaesthetized guinea pigs.

In the present study, we investigated neuronal responses to acoustic stimuli and cortical stimulation in the medial geniculate body (MGB) through in vivo intracellular recordings in anaesthetized guinea pigs. Of the 54 neurones examined with acoustic stimuli, 36 showed excitatory postsynaptic potential (EPSP) responses and 19 showed inhibitory postsynaptic potential (IPSP) responses to acoustic stimuli. Of the 36 EPSP neurones examined with corticofugal modulation, 29 received corticofugal depolarization, 3 corticofugal inhibition, and 4 showed no effect. Of the 19 IPSP neurones, 17 received corticofugal inhibition and 2 were not affected. The mean amplitude of the EPSPs evoked by acoustic stimuli was similar to that evoked by the electrical cortical stimulation (9.19 +/- 5.55 mV versus 9.22 +/- 5.16 mV). There was a significant correlation between the parameters of the EPSPs evoked by an acoustic stimulus and those evoked by cortical stimulation. The mean amplitude of the IPSP evoked by electrical cortical stimulation was significantly greater than that evoked by acoustic stimuli (11.6 +/- 3.8 mV versus 9.1 +/- 3.7 ms, P < 0.05). Seven auditory EPSP and 7 IPSP neurones were examined with corticofugal modulation and labelled with Neurobiotin. Of the 7 EPSP neurones, 5 showed excitatory responses to cortical stimulation and 2 demonstrated no effects. Four of the 5 neurones that received corticofugal depolarization were located in the lemniscal MGB and 1 in the non-lemniscal MGB; of the remaining 2, 1 was located in the lemniscal and the other in the non-lemniscal MGB. Of the 7 IPSP neurones, 1 received an excitatory corticofugal input followed by an inhibitory input and 4 received only an inhibitory corticofugal input, while the remainder demonstrated no corticofugal effects. All 7 neurones were located in the non-lemniscal MGB. The result that both ascending and descending inputs caused similarly shaped EPSPs reflects a neuronal endogenous characteristic irrespective of the physical locations of the synapses. The IPSP responses to both acoustic stimuli and electrical cortical stimulation are likely to be caused by feedback from the thalamic reticular nucleus.

Acoustic Stimulation↗

Submaximal stimuli activate different nerve fiber populations at different sites.

Theoretically, the largest and fastest nerve fibers are preferentially stimulated with submaximal stimuli. However, it is also well known that intraneural fascicular topography changes substantially along a proximal to distal axis. Because of this change in fascicular topography, we hypothesized the percutaneous submaximal stimuli applied to a nerve at different locations would stimulate different subpopulations of large fibers. We performed a series of collision studies by stimulating the ulnar nerve submaximally at proximal and distal sites at varying levels of stimulation intensity from motor threshold to supramaximal stimulation. The results suggest that variation in intraneural topography at different sites allows different large diameter nerve fiber subpopulations to be activated at submaximal stimuli, and emphasizes the importance of supramaximal stimulation to determine a valid conduction velocity.

Action Potentials↗

Evaluation of the functional state of the somato-motor system using SEP and interfering stimuli.

The analysis of median nerve stimulated SEPs recorded during interfering stimuli like hand movements, provides a promising tool to illuminate the functional state of the somato-motor system. Prior studies in general observed altered amplitudes of frontal (N30) and/or parietal (P27) peaks caused by interfering stimuli, demonstrating functional changes of the somato-motor system. We performed a study to evaluate whether different interfering stimuli cause different patterns of SEP alterations. SEPs were recorded with 64-channels and analysed using source analysis, global field power and mapping. Interfering stimuli were: mental complex finger movements, acted simple and acted complex finger movements, explorative finger movements and brushing of fingers I to III. Statistical evaluation revealed: 1) No difference between median nerve SEPs at rest and during mental finger movements. 2) Different patterns of maps computed at maxima of global field power between 25 and 35 ms post stimulus clearly distinguished between the 3 types of interfering stimuli, finger movements, explorative movements and pure sensory interfering stimulation in 8 out of 10 subjects. Hence, the analysis demonstrated different functional states of the somato-motor system to motor, pure sensory and both motor and sensory inputs.

Adult↗

[Participation of the visual projection and association cortical inputs in the formation of responses of neostriatal responses to visual stimuli in the alert cat].

Extracellular responses of caudate neurons to local light stimuli and to electrical stimulation of specific (area 17) and associative (the Clare-Bishop area) cortical zones were studied in awake cats. Stimulation of the Clare-Bishop area was found to be more effective for caudate cell responses in comparison with stimulation of area 17; it influenced more caudate cells (47%), whereas responses to area 17 stimulation occurred only in 8% of investigated cells. The mean latency values also were lower in the first case. Only a few neurons responded to stimulation of the both areas. Responses to visual stimuli and to electrical stimulation of these cortical areas were compared in the same caudate cells. Neurons which responded to visual stimuli were also better activated by the Clare-Bishop stimulation than by stimulation of area 17. Such cells were distributed preferably in the body of caudate nucleus. The possible role of the Clare-Bishop area in transmission of visual information to neostriatum is discussed.

Animals↗

Perceptual integration of intramuscular electrical stimulation in the focal and the referred pain area in healthy humans.

The aim of the study was to investigate the perceptual integration of simultaneous stimulation in a focal and a referred pain area to investigate whether referred pain is mainly caused by facilitation of on-going input from the referred pain area by stimulation in the focal pain area or if referred pain is a consequence of misinterpretation of the origin of inputs from the focal pain area. Pain was induced in twelve healthy individuals by intramuscular electrical stimulation in the left infraspinatus muscle (MI) or the left dorsolateral upper arm (UA), i.e. the area of referral commonly reported from stimulation in MI. Conditioning stimulation consisted of, in a counterbalanced order, no stimulation (baseline) and pain intensity rated as 2/10 and 4/10, respectively, on a category scale. During conditioning stimulation in MI, sensitivity to test stimuli was assessed in UA and vice versa. The test stimuli consisted of i.m. electrical stimulation corresponding to the perception threshold to innocuous electrical stimulation, the electrical pain threshold (EPT), and pain intensity rated as 2/10, 4/10 and 6/10, respectively. Conditioning stimulation corresponding to 2/10 did not result in a statistically significant change in sensitivity to any test stimuli in either location. During conditioning stimulation corresponding to 4/10 in m. infraspinatus, all twelve subjects reported referred pain in the dorsolateral upper arm. Compared to baseline, EPTs decreased in the referred pain area (P<0.001), while no other statistically significant changes in sensitivity to test stimuli were seen. Conditioning stimulation corresponding to 4/10 in the dorsolateral upper arm gave rise to referred pain in one individual (area of m. biceps brachii), and no statistically significant changes were seen in the sensitivity to electrical stimuli in m. infraspinatus. In conclusion, an effect at pain threshold level only was documented during simultaneous stimulation in the focal and referred pain area, which does not support facilitation of inputs from the referred pain area as the main mechanism generating referred pain. Instead, referred pain is most likely a consequence of misinterpretation of the origin of input from the stimulated focal pain area, due to excitation of neurones somewhere along the neuroaxis with projected fields in the referred pain area. The fact that conditioning stimulation in m. infraspinatus generated referred pain in the dorsolateral upper arm, but not vice versa suggests that the divergence of the input is not reciprocally arranged.

Adult↗

Identification of an N-terminally truncated form of the chemokine RANTES and granulocyte-macrophage colony-stimulating factor as major eosinophil attractants released by cytokine-stimulated dermal fibroblasts.

Eosinophil (Eo) granule proteins and, rarely, intact Eos represent a characteristic histopathologic feature of the dermal part of affected tissue in atopic dermatitis and some allergic reactions. Dermal fibroblasts are a rich source of cytokines and inflammatory mediators; therefore, we have investigated whether these cells release Eo chemoattractants when stimulated with different stimuli. Eo-chemotactic activity was detected after stimulation of cells with TNF-alpha and IL-1, but not when phorbol ester, PHA, or medium alone was used. Biochemical characterization of Eo-chemotactic activity in supernatants of NF-alpha-stimulated cells revealed both heparin-binding and nonbinding activity. HPLC purification with subsequent N-terminal sequencing and mass spectrometric analysis showed that the heparin-binding Eo-chemotactic peak corresponded to the chemokine [Tyr-RANTES]66 that also contained [Ser-RANTES]68 as contaminant, whereas the nonheparin-binding activity was identified as granulocyte-macrophage CSF (GM-CSF) by the use of neutralizing Abs. [Tyr-RANTES]66 was found to show identical behavior in the chemotaxis assay system with respect to potency and efficacy as natural [Ser-RANTES]68. These findings support the hypothesis that dermal fibroblasts can play an important role in the recruitment of Eo by release of the chemokine RANTES together with GM-CSF.

Amino Acid Sequence↗

The digastric reflex evoked by tooth-pulp stimulation in the cat and its modulation by stimuli applied to the limbs.

The digastric reflex evoked by electrical stimulation of tooth pulp in anaesthetized cats was studied together with the effects on this reflex of stimulating other parts of the body. The threshold for the digastric reflex generally lay in the range of stimulus intensities which would excite a large proportion of the pulpal afferent fibres which suggested that a large amount of central summation was required to evoke the reflex. During the course of 25/27 experiments, the threshold for the reflex increased. It was also found that repeated application of suprathreshold stimuli produced first an increase and then a decrease in the reflex response. The application of noxious but not of non-noxious mechanical conditioning stimuli to the limbs produced strong, long-lasting depressions of the digastric reflex. Electrical conditioning stimuli applied to the limbs also depressed the reflex; this depression had a latency of onset of 20-50 ms and lasted for up to 500 ms. When conditioning stimuli were applied to the saphenous nerve, the depression of the reflex occurred only when the stimuli were of an intensity sufficient to excite fibres conducting at less than 40 m X s-1; it may be assumed that some of these fibres would have been high threshold mechanoreceptors or nociceptors. These results show that noxious stimulation of anatomically remote structures can depress the activity of a population of trigeminal brainstem neurones. The opiate antagonist, naloxone, had no detectable effect on either the digastric reflex or the depression of the reflex produced by stimulating other parts of the body. The serotonin antagonists, methysergide and cinanserin, strongly depressed the digastric reflex but it was not clear whether these drugs also affected the depression of the reflex by the conditioning stimuli.

Animals↗

Corneal and conjunctival sensitivity to air stimuli.

AIMS: To determine the repeatability of ocular surface sensitivity to mechanical stimulation using air stimuli and the effect of contact lens (CL) wear on sensitivity. METHODS: Repeatability: 14 subjects (24-39 years) participated. Mechanical sensitivity to warmed (34 degrees C) and ambient (20 degrees C) air was measured for the central cornea (CC), inferior cornea (IC), and inferior conjunctiva (ICON). Measurements were taken on 12 days; six morning and six afternoon measurements. Differences between sites, time of day, and stimulus temperature were evaluated. CL wear: 10 subjects (22-30 years) participated. Measurements were taken at the same time of day, either following no wear, wear of a CL of oxygen permeability [Dk] of 28 x 10(-9) [cm/s][ml O(2)/ml mm Hg] or wear of a CL of Dk 140 x 10(-9) [cm/s][ml O(2)/ml mm Hg]. Differences between sites and wear conditions were evaluated. RESULTS: Repeatability: Sensitivity varied between sites (p<0.01), time of day (p<0.05), and stimulus temperatures (p<0.01). There were no significant differences between days. Mean thresholds for eye temperature stimuli were; CC 64.4 (SD 28.6) ml/min; IC 84.6 (40.0) ml/min; ICON 120.6 (40.4) ml/min and for ambient temperature stimuli were CC 53.9 (16.0) ml/min, IC 59.0 (20.0) ml/min; ICON 72.6 (43.7) ml/min. CL wear: Sensitivity varied between sites and wear conditions (p<0.05). Conjunctival sensitivity was increased after wear of highly oxygen permeable CLs but unaffected by wear of low oxygen permeable CLs. CONCLUSIONS: The prototype gas aesthesiometer is able to repeatably measure ocular surface sensitivity and measurements are consistent with previously reported techniques.

Adult↗

Origin of the facial long latency responses elicited by magnetic stimulation.

With magnetic stimulation (MS) it is possible to elicit bilateral long latency facial motor responses (LLRs). Due to a relatively wide magnetic field, the site of neural activation may take place in many different structures. The purpose of this study was to determine the site of origin of facial LLRs. The motor long latency responses were recorded bilaterally on the naso-labial folds (NLFs) with reference electrodes on the nose, and on some subjects also with reference electrodes on the chin. The stimulating coil was placed in the right parietal area. LLRs obtained with MS were compared to LLRs elicited electrically at the right stylomastoid foramen, supraorbital foramen, as well as cutaneous sensory area V1 of the trigeminal nerve. In addition, right sided high intensity electrical stimuli, paired magnetic stimulation and electrical stimulation with interstimulus intervals ranging from 0 to 80 msec were also applied for comparison. LLRs recorded with reference to the nose were always elicitable with MS as well as with the other stimulation procedures. The responses elicited with MS did not differ from those elicited electrically at various extracranial stimulation sites. With paired stimuli the second LLRs were inhibited by the preceding stimulation, whether given magnetically or electrically. In subjects with elicitable LLRs with chin references, the responses were always bilateral. Based on the similar characteristics with extracranial electrical stimuli, bilateral distribution of the responses, and inhibition of the second response with paired stimuli, it is concluded that the neural origin of LLRs to MS is in the extracranial trigeminal or facial nerve branches.

Adult↗

Baroreceptor reflex during arousal induced by electrical stimulation of the amygdala or by natural stimuli.

Baroreceptor control of heart rate was studied in 14 unanaesthetized cats during rest and arousal induced by either electrical stimulation of the central nucleus of the amygdala or by natural stimuli. The baroreceptor reflex was elicited by i.v. injections of angiotensin II and the sensitivity of the vagal component of the reflex expressed as the regression coefficient of the relationship between the systolic blood pressures of successive arterial pulses and their pulse intervals. Baroreceptor reflex sensitivity was reduced by all types of arousing procedure studied; an effect assumed to be a part of integrated cardiovascular adjustments accompanying arousal. Further studies in slightly anaesthetized cats were performed in order to investigate the time-course of such changes in reflex sensitivity. Th inhibition of the vagal component of the reflex did not last for the entire period of the electrically elicited arousal and it is suggested, therefore, that this might reflect the activation of mechanisms which limit orienting behaviour. Overall, these results indicate that the central nucleus of the amygdala may be involved in a transient integration of cardiovascular and behavioural responses during arousal.

Amygdala↗

Pacemaker neurons of the forebrain medical septal area and theta rhythm of the hippocampus.

The hippocampus is of critical importance for the organization of selective attention and memory. The activity of its neurons is rhythmically modulated by the direct afferent input from cholinergic and GABAergic neurons of the medial septal nucleus and the nucleus of diagonal band (MS-DB). This modulation is expressed in the hippocampal electroencephalogram as a slow (4-9 Hz) sinusoidal theta rhythm. The paper presents a short review of the authors' data on the properties of MS-DB cells, their interactions in organizing the theta expression in the hippocampus, and the probable functional significance of this rhythm. Extracellular recordings of neural activity of the MS-DB and hippocampus were performed in chronic alert rabbits with transection of various afferent pathways, in brain slices, and in the embryonic tissue grafts developing in the anterior eye chamber and brain. Theta modulation disappeared in all structures deprived of the MS-DB afferent connections, but was retained by MS-DB neurons under isolation conditions. A limited group of MS-DB neurons (6-8%) discharge in regular bursts after complete synaptic blockade in high Mg(2+)-low Ca2+ medium, while other neurons in these conditions exhibited the highly regular single-spike activity. Thus, the MS-DB neurons can be regarded as bursting and regular pacemakers. The frequency of bursts and the number of neurons involved in the rhythmic activity are in parallel increased by afferent stimulation-either natural (sensory stimuli) or imitated by electrical stimulation of ascending reticular formation. An increase in the number of MS-DB neurons secondarily involved in theta bursts with the resulting expression of theta rhythm in EEG can be also obtained by up-regulation of endogenous acetylcholine (by physostigmine), but in this case the frequency of theta is not changed. The MS-DB neurons contain acetylcholine and GABA as neurotransmitters. Analysis of the effects of their agonists and antagonists suggests that the frequency of theta depends on the GABAergic mechanism, while its power is controlled by cholinergic influences. In the control state, input signals triggered in the hippocampal neurons an inhibitory reset followed by synchronous theta modulation, gradually habituating during repeated presentations of the stimulus. Against the background of continuous theta evoked by physostigmine, the responses were blocked or significantly depressed, while after theta suppression by scopolamine the efficacy of the stimuli was increased, and habituation was absent. It is suggested that the theta rhythm operates as a selective filter: it augments and prolongs the input signal by which it was triggered and simultaneously protects it from the interference of extraneous stimuli appearing during its processing and registration. The theta rhythm may be regarded as an important mechanism of selective attention, which is a prerequisite for memory trace formation.

Animals↗

Differential effects of transcranial magnetic stimulation of left and right posterior parietal cortex on mental rotation tasks.

A recently published study used the interference strategy of transcranial magnetic stimulation (TMS) to demonstrate the role of the right posterior parietal cortex (PPC) in the mental rotation of alphanumeric stimuli. We used similar stimulation parameters over the same left and right PPC regions, and examined the ability to rotate more complex 3D Shepard and Metzler (1971) images. There was reduced accuracy of performance with both right and left PPC stimulation for different angles of rotation of the visual stimuli. Right PPC stimulation led to reduced accuracy to rotate stimuli by 1200, whereas left PPC stimulation affected 180 degrees C rotation. We hypothesise that the two hemispheres make different contributions to the processing underlying visuospatial mental imagery: the right PPC is important for spatial rotations through smaller angles; the left hemisphere has a unique role when the stimuli to be compared are rotated through 180 degrees C, a task that engages verbal strategies due to the well-documented special nature of enantiomorphs.

Adult↗

A pharmacokinetic-pharmacodynamic model for neuromuscular blocking agents to predict train-of-four twitches.

The train-of-four (TOF) stimulation pattern consists of 4 stimuli (T1, T2, T3, and T4) at 2 Hz, and is used in daily anesthesiological practice to determine the degree of relaxation caused by muscle relaxants. At a surgical levels of relaxation the degree of relaxation can be estimated by counting the number of "measurable" or "visible" muscular reactions to the 4 stimuli in the TOF stimulation pattern (TOF count). During recovery relaxation can be estimated by calculating the TOF ratio (T4/T1). Bartkowski and Epstein described a pharmacokinetic-pharmacodynamic (PK-PD) model to predict TOF ratio by modifying and extending the PK-PD model as described by Sheiner to use a hypothetical distributed effect compartment described by a median equilibration rate constant and a dispersion parameter. We extended the Bartkowski and Epstein PK-PD model to simulate all four TOF twitches by including EC50 terms for T2 and T3. We fit this model to data from the pig and compared the results to fitted models using separate PD models for each TOF twitch (extended Sheiner model). The extended Bartkowski and Epstein model fit the twitch height data from all four TOF twitches better than the extended Sheiner model and has fewer parameters.

Anesthesia↗

Dexmedetomidine modulates cardiovascular responses to stimulation of central nervous system pressor sites.

UNLABELLED: Halothane attenuates the alterations in arterial pressure (BP) and heart rate (HR) produced by central nervous svstem (CNS) stimulation. We examined the effects of the alpha2-adrenergic agonist dexmedetomidine, with and without halothane, on cardiovascular regulation during CNS pressor site stimulation in chronically instrumented cats. Stimuli trains via bipolar stimulating electrodes in the hypothalamus and reticular formation elicited pressor responses. Dexmedetomidine-induced (15 microg/kg PO) bradycardia was greater in the presence of halothane. CNS stimulation increased BP and HR, which were dose-dependently attenuated by halothane (hypothalamic stimulation 71 +/- 9 mm Hg at control, 25 +/- 5 and 15 +/- 3 mm Hg at 1.0% and 1.5% halothane, respectively). Although dexmedetomidine alone did not alter pressor responses, halothane plus dexmedetomidine attenuated pressor responses in a potentially synergistic fashion (hypothalamic stimulation 67 +/- 8 mm Hg at control, 2 +/- 1 and 1 +/- 0.4 mm Hg at 1.0% and 1.5% halothane, respectively). These results suggest differences in the disruptive effects of CNS-mediated cardiovascular responses by halothane and dexmedetomidine, and that dexmedetomidine has an anesthetic-sparing effect on these CNS-mediated cardiovascular control mechanisms, potentiating the depressant effect of halothane. IMPLICATIONS: A new potential anesthetic adjunct, dexmedetomidine, does not attenuate brain-mediated increases in blood pressure, but the combination of dexmedetomidine and the anesthetic halothane acts to modulate central cardiovascular responses.

Adrenergic alpha-Agonists↗

Perceptual and response bias in visuospatial neglect due to frontal and parietal repetitive transcranial magnetic stimulation in normal subjects.

Recently some authors have challenged the conventional association of directional motor neglect with damage of frontal structures, showing that pure sensory perceptual neglect (classically associated with parietal lesion) can follow damage of right frontal cortex. The aim of the present study was to assess the type of defect in visuo-spatial attention consequent upon a virtual frontal or parietal lesion induced by transcranial magnetic stimulation in normal subjects. To this purpose eleven subjects performed a visuo-spatial task requiring judgement about the length of the two segments of asymmetrically bisected horizontal lines, presented for 50 ms on a computer monitor. After each visual stimulus, subjects made a binary forced choice decision according to two different response conditions: A and B. In condition A, they had to name (right or left) the longer segment and in B the shorter segment of the line. The task was given in baseline condition and during repetitive transcranial magnetic stimulation. Trains of 10 stimuli at 25 Hz of frequency were applied over right frontal premotor and right posterior parietal areas, synchronously with visual stimuli. Parietal and frontal magnetic stimulation gave rise to significant perceptual bias as compared to baseline performance (i.e. subjects made opposite errors in the two response conditions). No significant response bias (i.e. the tendency to name the same side of the line in the two response conditions) was induced by magnetic stimulation on parietal and frontal sites. The present study highlights both the relevant contribution of frontal cortex in the determinism of neglect and the predominant role of sensory perceptual factors in parietal and frontal neglect.

Adult↗

Tension development by isolated muscle spindles of the cat.

1. Tension developed by mammalian intrafusal muscle fibres stimulated directly with single or repetitive electrical pulses has been determined in sixteen isolated lumbrical muscle spindles of the cat's foot.2. On microscopic observation, contraction to single shocks was observed in ten preparations. In three of these preparations twitch-like records were obtained.3. In the preparations that ;twitched', the tetanic fusion frequency was 10 pulses/sec at 20 degrees C.4. In the ;twitching' intrafusal fibres the magnitude of contraction increased with increase of stimulus strength.5. In one of these spindles, only ;slow' contractions were obtained at low stimulus strengths when it was stimulated repetitively at 100 pulses/sec. With increase in stimulus strength, such ;slow' contractions gave twitch-like responses towards the end of a period of repetitive stimulation.6. Tension changes were recorded from all preparations stimulated repetitively. With supramaximal stimuli, the usual frequency of stimulation at which intrafusal tension was recorded was 10 pulses/sec at 20 degrees C. The magnitude of contraction increased progressively with increase in stimulus frequency.7. Larger intrafusal tensions and faster time courses of contraction were obtained when the electrodes were astride the capsular region than when they were over the inferior extracapsular region.

Journal Article↗