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Biomedical subjects

J M Walker

Publications and source records attributed to J M Walker.

At least 37 records · Page 2Linked to original sources
PubMed2000

Cerebral microembolism detected by transcranial Doppler during cardiac procedures.

BACKGROUND AND PURPOSE: Cerebral embolism with clinical sequelae may rarely complicate cardioversion and cardiac catheterization. Transcranial Doppler sonography has recently been introduced to monitor microemboli entering the middle cerebral artery in cardiac and carotid surgery. We therefore used this technique to evaluate the risk of asymptomatic embolism during common cardiac procedures. METHODS: Patients were monitored by transcranial Doppler while undergoing direct current cardioversion (n=15) and cardiac catheterization (n=17). RESULTS: Microemboli were detected in all patients having cardiac catheterization but in only 1 patient after cardioversion. CONCLUSIONS: Microembolism occurred frequently during cardiac catheterization and rarely during cardioversion. It is not yet known whether this has clinical relevance.

Cardiac Catheterization↗
PubMed2000

Nasal continuous positive airway pressure devices do not maintain the set pressure dynamically when tested under simulated clinical conditions.

STUDY OBJECTIVES: Nasal continuous positive airway pressure (CPAP) is standard therapy for obstructive sleep apnea syndrome. The effective nasal mask pressure may be adversely affected by factors that increase system resistance (eg, long tubing and/or water condensation) and by dynamic variables (breathing frequency [f] and tidal volume [VT]). The present study was conducted in order to assess the performance of CPAP machines throughout a range of simulated clinical conditions. DESIGN: Four currently used CPAP machines were tested at settings of 5, 10, 15, and 20 cm H(2)O using a pulmonary waveform generator to produce VTs of 0.4, 0.8, and 1.2 L at frequencies of 10, 20, and 30 breaths/min. Machines were tested under five conditions: 6-foot and 12-foot tubing, with and without an in-line humidifier, and 12-foot tubing with humidifier and water condensation. MEASUREMENTS: Maximum and minimum mask pressure measurements were obtained during five respiratory cycles for each dynamic variable under each of the five conditions and CPAP settings (180 experiments on each of four CPAP models). RESULTS: Using typical clinical parameters (VT, 0.4 L and 0.8 L; f, 10 breaths/min and 20 breaths/min; and CPAP, 5 to 15 cm H(2)O), mask pressure consistently varied above and below the set point when additional tubing and/or a humidifier were added to the system (0.7 to 2.9 cm H(2)O below and 0.5 to 1.0 cm H(2)O above the set pressure). Water condensation caused large pressure deviations (inspiratory pressure ranged from 3.5 to 5.6 cm H(2)O below set pressure, and expiratory pressure ranged from 0.7 to 3.5 cm H(2)O above set pressure). CONCLUSIONS: Therapy and compliance could be adversely affected because some CPAP machines in current use do not maintain constant continuous mask pressure when tested using simulated conditions, especially when water condenses in the tubing.

Equipment Design↗
PubMed1999

Pain modulation by release of the endogenous cannabinoid anandamide.

Synthetic cannabinoids produce behavioral analgesia and suppress pain neurotransmission, raising the possibility that endogenous cannabinoids serve naturally to modulate pain. Here, the development of a sensitive method for measuring cannabinoids by atmospheric pressure-chemical ionization mass spectrometry permitted measurement of the release of the endogenous cannabinoid anandamide in the periaqueductal gray (PAG) by in vivo microdialysis in the rat. Electrical stimulation of the dorsal and lateral PAG produced CB1 cannabinoid receptor-mediated analgesia accompanied by a marked increase in the release of anandamide in the PAG, suggesting that endogenous anandamide mediates the behavioral analgesia. Furthermore, pain triggered by subcutaneous injections of the chemical irritant formalin substantially increased the release of anandamide in the PAG. These findings indicate that the endogenous cannabinoid anandamide plays an important role in a cannabinergic pain-suppression system existing within the dorsal and lateral PAG. The existence of a cannabinergic pain-modulatory system may have relevance for the treatment of pain, particularly in instances where opiates are ineffective.

Analgesia↗
PubMed1999

Anatomical basis for cannabinoid-induced antinociception as revealed by intracerebral microinjections.

Cannabinoids suppress behavioral and neurophysiological responses to noxious stimuli in rodents when administered systemically. The purpose of this study was to extend previous studies of the site of cannabinoid analgesia. Rats were tested in the tail flick test before and after microinjections of the cannabinoid agonist WIN55, 212-2 (5 microg) into one of 17 different brain regions. WIN55,212-2 significantly elevated tail-flick latencies when injected into the amygdala, the lateral posterior and submedius regions of the thalamus, the superior colliculus and the noradrenergic A5 region. By contrast, pain behavior was unaffected by microinjections of the cannabinoid into the other 11 areas examined (prefrontal cortex, nucleus accumbens, lateral hypothalamus, substantia nigra, cuneiform nucleus, anterior pretectal, intralaminar, parafasicular, posterior, thalamic nuclei, as well as the ventral medial, ventral lateral nuclei in the posterior thalamus).

Amygdala↗
PubMed1999

Duplication of 7p12.1-p13, including GRB10 and IGFBP1, in a mother and daughter with features of Silver-Russell syndrome.

Silver-Russell syndrome (SRS) has been associated with maternal uniparental disomy (UPD) of chromosome 7 in approximately 10% of cases, suggesting that at least one imprinted gene on chromosome 7 is involved in the pathogenesis of the disease. We report a proximal 7p interstitial inverted duplication in a mother and daughter both of whom have features of SRS, including marked short stature, low birth weight, facial asymmetry and 5th finger clinodactyly. Fluorescence in situ hybridisation (FISH) with YAC probes enabled delineation of the duplicated region to 7p12.1-p13. This region of proximal chromosome 7 is known to be homologous to an imprinted region in the mouse chromosome 11 and contains the growth-related genes GRB10 (growth factor receptor-bound protein 10), EGFR (epidermal growth factor receptor) and IGFBP1 (insulin-like growth factor binding protein 1), all of which have been suggested as candidate genes for SRS. Molecular analysis showed that the duplication in both mother and daughter spanned a distance of approximately 10 cM and included GRB10 and IGFBP1 but not EGFR. The de novo duplication in the proband's mother was shown to be of paternal origin. In order to test the hypothesis that sub-microscopic duplications of 7p, whether maternal or paternal in origin, are responsible for at least some cases of SRS, we screened a further eight patients referred to our laboratory for SRS. None were found to have duplications of either GRB10 or IGFBP1. The hypothesis that sub-microscopic duplications including GRB10 and IGFBP1 is a cause of SRS remains a possibility and warrants further investigation. Importantly, in contrast to current thinking, our results suggest that imprinted genes may not underlie the SRS phenotype, and we propose an alternative hypothesis to explain the occurrence of maternal UPD 7 seen in some cases of SRS.

Abnormalities, Multiple↗
PubMed1999

The neurobiology of cannabinoid analgesia.

The discovery of cannabinoid receptors and their putative endogenous ligands raises questions as to the nature of the effects produced by cannabinoids on neural circuits that mediate pain and whether endogenous cannabinoids produced by the brain or in the periphery serve naturally to modulate pain. A sizable body of previous work showed that cannabinoid agonists suppress pain behavior in a variety of models of acute and chronic pain. However, at appropriate doses, cannabinoids also profoundly suppress motor behavior (see Sañudo-Peña et al., this volume), which complicates the interpretation of behavioral analgesia since a motor response is the endpoint of virtually all such studies. Studies conducted in this laboratory used biochemical and neurophysiological measures to determine whether cannabinoids suppress nociceptive neurotransmission. The results showed that cannabinoids suppress nociceptive neurotransmission at the level of the spinal cord and the thalamus. These effects are reversible, receptor mediated, selective for painful as opposed to nonpainful somatic stimuli, and track the behavioral analgesia both in time course and potency.

Analgesia↗
PubMed1999

Motor actions of cannabinoids in the basal ganglia output nuclei.

The levels of CB1 cannabinoid receptors in the basal ganglia are the highest in the brain, comparable to the levels of dopamine receptors, a major transmitter in the basal ganglia. This localization of receptors is consistent with the profound effects on motor function exerted by cannabinoids. The output nuclei of the basal ganglia, the globus pallidus (GP) and substantia nigra reticulata (SNr), apparently lack intrinsic cannabinoid receptors. Rather, the receptors are located on afferent terminals, the striatum being the major source. Cannabinoids blocked the inhibitory action of the striatal input in the SNr. Furthermore, cannabinoids blocked the excitatory effect of stimulation of the subthalamic input to the SNr revealing, along with data from in situ hybridization studies, that this input is another likely source of cannabinoid receptors to the SNr. Similar actions of cannabinoids were observed in the GP. Behavioral studies further revealed that the action of cannabinoids differs depending upon which input to the output nuclei of the basal ganglia is active. The inhibitory striatal input is quiescent and the cannabinoid action is observable only upon stimulation of the striatum, while the noticeable effect of cannabinoids under basal conditions would be on the tonically active subthalamic input. These data suggest that the recently discovered endogenous cannabinergic system exerts a major modulatory action in the basal ganglia by its ability to block both the major excitatory and inhibitory inputs to the SNr and GP.

Animals↗
PubMed1999

Cannabinoid CB1 receptors are localized primarily on cholecystokinin-containing GABAergic interneurons in the rat hippocampal formation.

Localization of cannabinoid CB 1 receptors on GABAergic interneurons in the rat hippocampal formation was studied by double-labeling immunohistochemistry with confocal microscopy. Virtually all CB1-immunoreactive neurons (95%) are GABAergic. CB 1 fluorescence showed a punctate pattern. In contrast, the GABA fluorescence was distributed homogeneously, suggesting that while CB 1 receptors and GABA exist in the same cells they are not localized in the same subcellular compartments. Although virtually all CB1 neurons were GABAergic, many GABAergic neurons did not contain CB1 receptors. GABAergic interneurons in the hippocampal formation can be further divided into subpopulations with distinct connections and functions, using cell markers such as neuropeptides and calcium binding proteins. CB1 receptors were highly co-localized with cholecystokinin and partially co-localized with calretinin and calbindin, but not with parvalbumin. This suggests that cannabinoids may modulate GABAergic neurotransmission at the synapses on the soma and at synapses on the proximal dendrites of the principal neurons, as well as at synapses on other GABAergic interneurons.

Animals↗
PubMed1999

Sequences within both the N- and C-terminal domains of phytochrome A are required for PFR ubiquitination and degradation.

Photoconversion of the plant photoreceptor phytochrome A (phyA) from its inactive Pr form to its biologically active Pfr from initiates its rapid proteolysis. Previous kinetic and biochemical studies implicated a role for the ubiquitin/26S proteasome pathway in this breakdown and suggested that multiple domains within the chromoprotein are involved. To further resolve the essential residues, we constructed a series of mutant PHY genes in vitro and analyzed the Pfr-specific degradation of the resulting photoreceptors expressed in transgenic tobacco. One important site is within the C-terminal half of the polypeptide as its removal stabilizes oat phyA as Pfr. Within this half is a set of conserved lysines that are potentially required for ubiquitin attachment. Substitution of these lysines did not prevent ubiquitination or breakdown of Pfr, suggesting either that they are not the attachment sites or that other lysines can be used in their absence. A small domain just proximal to the C-terminus is essential for the form-dependent breakdown of the holoprotein. Removal of just six amino acids in this domain generated a chromoprotein that was not rapidly degraded as Pfr. Using chimeric photoreceptors generated from potato PHYA and PHYB, we found that the N-terminal half of phyA is also required for Pfr-specific breakdown. Only those chimeras containing the N-terminal sequences from phyA were ubiquitinated and rapidly degraded as Pfr. Taken together, our data demonstrate that, whereas an intact C-terminal domain is essential for phyA degradation, the N-terminal domain is responsible for the selective recognition and ubiquitination of Pfr.

Amino Acid Sequence↗
PubMed1999

Use of ubiquitin fusions to augment protein expression in transgenic plants.

A major goal of plant biotechnology is the production of genetically engineered crops that express natural or foreign proteins at high levels. To enhance protein accumulation in transgenic plants, we developed a set of vectors that express proteins and peptides as C-terminal translational fusions with ubiquitin (UBQ). Studies of several proteins in tobacco (Nicotiana tabacum) showed that: (a) proteins can be readily expressed in plants as UBQ fusions; (b) by the action of endogenous UBQ-specific proteases (Ubps), these fusions are rapidly and precisely processed in vivo to release the fused protein moieties in free forms; (c) the synthesis of a protein as a UBQ fusion can significantly augment its accumulation; (d) proper processing and localization of a protein targeted to either the apoplast or the chloroplast is not affected by the N-terminal UBQ sequence; and (e) single amino acid substitutions surrounding the cleavage site can inhibit in vivo processing of the fusion by Ubps. Noncleavable UBQ fusions of beta-glucuronidase became extensively modified, with additional UBQs in planta. Because multiubiquitinated proteins are the preferred substrates of the 26S proteasome, noncleavable fusions may be useful for decreasing protein half-life. Based on their ability to augment protein accumulation and the sequence specificity of Ubps, UBQ fusions offer a versatile way to express plant proteins.

Amino Acid Sequence↗
PubMed1999

Cannabinoid suppression of noxious heat-evoked activity in wide dynamic range neurons in the lumbar dorsal horn of the rat.

The effects of cannabinoid agonists on noxious heat-evoked firing of 62 spinal wide dynamic range (WDR) neurons were examined in urethan-anesthetized rats (1 cell/animal). Noxious thermal stimulation was applied with a Peltier device to the receptive fields in the ipsilateral hindpaw of isolated WDR neurons. To assess the site of action, cannabinoids were administered systemically in intact and spinally transected rats and intraventricularly. Both the aminoalkylindole cannabinoid WIN55,212-2 (125 microg/kg iv) and the bicyclic cannabinoid CP55,940 (125 microg/kg iv) suppressed noxious heat-evoked activity. Responses evoked by mild pressure in nonnociceptive neurons were not altered by CP55,940 (125 microg/kg iv), consistent with previous observations with another cannabinoid agonist, WIN55,212-2. The cannabinoid induced-suppression of noxious heat-evoked activity was blocked by pretreatment with SR141716A (1 mg/kg iv), a competitive antagonist for central cannabinoid CB1 receptors. By contrast, intravenous administration of either vehicle or the receptor-inactive enantiomer WIN55,212-3 (125 microg/kg) failed to alter noxious heat-evoked activity. The suppression of noxious heat-evoked activity induced by WIN55,212-2 in the lumbar dorsal horn of intact animals was markedly attenuated in spinal rats. Moreover, intraventricular administration of WIN55,212-2 suppressed noxious heat-evoked activity in spinal WDR neurons. By contrast, both vehicle and enantiomer were inactive. These findings suggest that cannabinoids selectively modulate the activity of nociceptive neurons in the spinal dorsal horn by actions at CB1 receptors. This modulation represents a suppression of pain neurotransmission because the inhibitory effects are selective for pain-sensitive neurons and are observed with different modalities of noxious stimulation. The data also provide converging lines of evidence for a role for descending antinociceptive mechanisms in cannabinoid modulation of spinal nociceptive processing.

Analgesics↗
PubMed1999

Cannabinoid WIN 55,212-2 inhibits the activity-dependent facilitation of spinal nociceptive responses.

Cannabinoids suppress nociceptive processing of acute stimuli, but little is known about their effects on processes that lead to hyperexcitability of nociceptive neurons following prolonged noxious stimulation. Wind-up, the increasingly strong response of spinal nociceptive neurons to repetitive noxious electrical stimuli, results from a fast-rising cumulative depolarization and increase in intracellular calcium concentration. These processes produce central sensitization, the increased excitability of spinal nociceptive neurons that contributes to the hyperalgesia and allodynia associated with chronic pain. Intravenous injection of the potent, synthetic cannabinoid agonist WIN 55, 212-2, but not the inactive enantiomer, WIN 55,212-3, dose-dependently decreased the wind-up of spinal wide dynamic range and nociceptive-specific neurons independent of acute responses to activation of low- and high-threshold primary afferents. This is the first direct evidence that cannabinoids inhibit the activity-dependent facilitation of spinal nociceptive responses.

Animals↗
PubMed1999

The effect of earplugs on sleep measures during exposure to simulated intensive care unit noise.

BACKGROUND: Sleep deprivation may contribute to impaired immune function, ventilatory compromise, disrupted thermoregulation, and delirium. Noise levels in intensive care units may be related to disturbed sleep patterns, but noise reduction has not been tested in this setting. OBJECTIVE: To measure the effect of a noise reduction intervention on the sleep of healthy subjects exposed to simulated intensive care unit noise. METHODS: After digital audiotape recording of noise and development of the noise reduction intervention, 5 nocturnal 8-hour periods of sleep were measured in 6 paid, healthy volunteers at 7-day intervals in a sleep disorders center. Polysomnographic data were collected by experienced sleep disorders technicians and scored by certified raters. After the first 3 quiet nights, earplugs were randomly assigned to be worn on the fourth and fifth nights during exposure to the recorded noise. Sound pressure levels were measured during all 5 nights. RESULTS: Sleep architecture and sound measurements on quiet nights did not differ significantly. Sound levels were significantly lower on quiet nights than on noise nights. Exposure to the noise increased the number of awakenings, percentage of stage 2 sleep, and rapid eye movement latency and decreased time asleep, sleep maintenance efficiency index, and percentage of rapid eye movement sleep. Earplugs worn during exposure to the noise produced a significant decrease in rapid eye movement latency and an increase in the percentage of rapid eye movement sleep. CONCLUSION: The results provide a reasonable basis for testing the effects of earplugs on the sleep of critically ill subjects.

Adult↗
PubMed1999

Liquid chromatographic-mass spectrometric measurement of the endogenous cannabinoid 2-arachidonylglycerol in the spinal cord and peripheral nervous system.

AIM: To develop a sensitive method for measuring the putative endocannabinoid 2-arachidonylglycerol (2-AG) in the peripheral and central nervous system. METHODS: A method using atmospheric pressure chemical ionization (APCI) liquid chromatography/mass spectrometry (LC/MS) was developed to determine the levels of 2-AG in methanol extracts of the rat lumbar spinal cord, dorsal root ganglion (DRG), and sciatic nerve. RESULTS: 2-AG was detected with high sensitivity and minimal sample preparation. The levels in the tissues analyzed were < or = pmol/mg wet weight. Similar levels were found in the spinal cord and the DRG, whereas approximately 7-fold lower levels were observed in the sciatic nerve. CONCLUSION: 2-AG is present in the peripheral nervous system, and the levels are markedly higher in cell bodies than those in axons.

Animals↗
PubMed1999

CB1 receptor localization in rat spinal cord and roots, dorsal root ganglion, and peripheral nerve.

AIM: The localization of CB1 receptors in the spinal cord, spinal roots, dorsal root ganglion (DRG), and peripheral nerve of the rat was determined. METHODS: We studied the distribution of CB1 cannabinoid receptors by immunohistochemistry using an antibody raised against the N-terminal of the receptor. RESULTS: The spinal cord showed numerous transverse fibers labelled for CB1 receptors throughout and concentrated in the dorsal horn. Lightly-stained cells were observed throughout the spinal cord gray matter. The DRG also showed cells and fibers labelled for CB1 receptors. Labelled fibers were observed in both dorsal and ventral roots as well as in peripheral nerves. CONCLUSION: The presence of CB1 receptors in the DRG, the dorsal root, and the dorsal horn is in accordance with the analgesic effects of cannabinoids. The presence of labelled cells and fibers in the ventral horn and ventral root provides a substrate for cannabinoid-induced muscle relaxant and antispastic effects.

Animals↗
PubMed1999

Dopaminergic system does not play a major role in the precipitated cannabinoid withdrawal syndrome.

AIM: To determine the dopaminergic system involvement in precipitated cannabinoid withdrawal syndrome. METHODS: The dopamine D1 receptor antagonist SCH23390 or the dopamine D2 receptor antagonist sulpiride was administered to rats chronically treated with either delta 9-tetrahydrocannabinol (THC) or vehicle. Subjects were then injected with either SR141716A or vehicle and behavior was observed for 1 h. RESULTS: Administration of the cannabinoid receptor antagonist SR141716A to animals chronically treated with THC as described by Tsou et al (1995) produced a profound withdrawal syndrome. Treatment with dopamine antagonists did not attenuate cannabinoid precipitated withdrawal syndrome in THC tolerant animals while the agonists increased the syndrome. CONCLUSION: It is unlikely that the dopaminergic system plays a major role in mediating the behavioral aspects of the cannabinoid withdrawal syndrome.

Animals↗