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Biomedical subjects

V Campbell

Publications and source records attributed to V Campbell.

At least 19 recordsLinked to original sources
PubMed2006

Influence of strong static magnetic fields on primary cortical neurons.

Intense uniform magnetic fields, such as those used in magnetic resonance imaging (MRI), are thought to exert little influence at the cellular level. Here we report modifications of the signaling cascades in rat cortical neurons cultured for 1 h in magnetic fields of up to 5 Tesla. The activation of c-Jun N-terminal kinase (JNK) increases monotonically with field strength, with a maximal activation of approximately 10% at 5 T, whereas the activation of extra cellular-regulated kinase (ERK) shows a maximum at 0.75 T ( approximately 10%). Since ERK is involved in cellular differentiation, these results indicate a magnetic induction of the signaling events associated with differentiation. However, the cells respond to further increases in field strength by evoking a stress response, since JNK is a stress-activated protein kinase. Three possible mechanisms are discussed and of these, the most plausible is magnetic field induced change in the membrane rest potential, a microscale magnetohydrodynamic effect. This mechanism most likely involves the activation of voltage dependent Ca(2+) channel opening; since intracellular Ca(2+) concentration was also found to be modified by the static magnetic field.

Animals↗
PubMed2001

Delta 9-tetrahydrocannabinol induces the apoptotic pathway in cultured cortical neurones via activation of the CB1 receptor.

Delta 9-tetrahydrocannabinol, the principal psychoactive component of marijuana, exerts a variety of effects on the CNS, including impaired cognitive function and neurobehavioural deficits. The mechanisms underlying these neuronal responses to tetrahydrocannabinol are unclear but may involve alterations in neuronal viability. Tetrahydrocannabinol has been shown to influence neuronal survival but the role of the cannabinoid receptors in the regulation of neuronal viability has not been fully clarified. In this study we demonstrate that tetrahydrocannabinol promotes the release of cytochrome c, activates caspase-3, promotes cleavage of the DNA repair enzyme poly-ADP ribose polymerase and induces DNA fragmentation in cultured cortical neurones. These effects of tetrahydrocannabinol were completely abrogated by the CB(1) receptor antagonist AM-251. The findings of this study demonstrate that tetrahydrocannabinol induces apoptosis in cortical neurones in a manner involving the CB1 subtype of cannabinoid receptor.

Animals↗
PubMed2000

Lipopolysaccharide inhibits long term potentiation in the rat dentate gyrus by activating caspase-1.

Lipopolysaccharide, a component of the cell wall of Gram-negative bacteria, may be responsible for at least some of the pathophysiological sequelae of bacterial infections, probably by inducing an increase in interleukin-1beta (IL-1beta) concentration. We report that intraperitoneal injection of lipopolysaccharide increased hippocampal caspase-1 activity and IL-1beta concentration; these changes were associated with increased activity of the stress-activated kinase c-Jun NH(2)-terminal kinase, decreased glutamate release, and impaired long term potentiation. The degenerative changes in hippocampus and entorhinal cortical neurones were consistent with apoptosis because translocation of cytochrome c and poly(ADP-ribose) polymerase cleavage were increased. Inhibition of caspase-1 blocked these changes, suggesting that IL-1beta mediated the lipopolysaccharide-induced changes.

Animals↗
PubMed2000

The role of ceramide in the modulation of intracellular Ca2+ levels by interleukin 1 beta in rat cortical synaptosomes.

We recently demonstrated that the pro-inflammatory cytokine, interleukin 1beta (IL-1beta) elevates intracellular free Ca2+ levels ([Ca2+]i) in rat cortical synaptosomes in a manner involving activation of the IL-1 receptor and stimulation of p42 mitogen-activated protein (MAP) kinase. We now report that the effects of IL-1beta on [Ca2+]i are mimicked by the sphingolipid metabolite ceramide. In cortical synaptosomes ceramide elevates [Ca2+]i in a p42 MAP kinase-dependent manner, and we conclude that the effects of IL-1beta on Ca2+ homeostasis involve ceramide as an upstream component of the p42 MAP kinase pathway.

Animals↗
PubMed1999

Effect of polyethylene occlusive skin wrapping on heat loss in very low birth weight infants at delivery: a randomized trial.

OBJECTIVE: Significant evaporative heat loss in the very low birth weight infant can occur in the delivery room. We investigated the effect of polyethylene wrap applied immediately at birth (without drying) on rectal temperature measured at nursery admission. STUDY DESIGN: Sixty-two consecutive infants delivered at <32 weeks' gestation were stratified by gestational age and randomly allocated to resuscitation with polyethylene wrap. All infants were resuscitated under radiant warmers. Wraps were removed on nursery admission. Rectal temperature was taken by digital electronic thermometer. RESULTS: Fifty-nine of 62 recruited infants completed the study. Maternal temperature, delivery room temperature, transfer-incubator temperature, and time to admission were recorded. Use of occlusive wrapping resulted in a significantly higher admission rectal temperature in infants <28 weeks' gestation (difference in means = 1.9 C, P <.001). No significant difference was seen in admission rectal temperature in infants of 28 to 31 weeks' gestation (difference in means = 0.17 C, P =.47). All 5 deaths were in the nonwrap group (vs wrap, P =.04); their mean temperature was 35.1 C versus 36.5 C in survivors (P =.001). CONCLUSIONS: Occlusive wrapping of very low birth weight infants at delivery reduces postnatal temperature fall. This may result in a decreased mortality rate.

Apgar Score↗
PubMed1998

Biphasic modulation of intracellular Ca2+ concentration by interleukin-1beta in cortical synaptosomes: involvement of a pertussis toxin-sensitive G-protein and mitogen-activated protein kinase.

The modulation of intracellular Ca2+ concentration ([Ca2+]i) by the pro-inflammatory cytokine interleukin-1beta (IL-1beta) was assessed in synaptosomes loaded with the Ca2+-sensitive dye, Fura-2AM. IL-1beta was found to exert a biphasic effect on the KCl-induced rise in [Ca2+]i, extending an inhibitory effect at lower (3.5 ng/ml) concentrations, and a stimulatory effect at high (100 ng/ml) concentrations. The inhibitory action of IL-1beta on [Ca2+]i was sensitive to pertussis toxin (PTX; 2 microg/ml), indicating a role for a PTX-sensitive G-protein, but was unaffected by the p42 mitogen-activated protein kinase kinase (MAP kinase kinase) inhibitor, PD 098059 (2 microM). In contrast, the stimulatory action of higher concentrations of IL-1beta on [Ca2+]i was blocked by PD 098059 and unaffected by PTX. We conclude that the biphasic actions of IL-1beta on the KCl-induced rise in [Ca2+]i are mediated through activation of alternative second messenger pathways.

Animals↗
PubMed1997

Voltage-dependent binding and calcium channel current inhibition by an anti-alpha 1D subunit antibody in rat dorsal root ganglion neurones and guinea-pig myocytes.

1. The presence of calcium channel alpha 1D subunit mRNA in cultured rat dorsal root ganglion (DRG) neurones and guinea-pig cardiac myocytes was demonstrated using the reverse transcriptase-polymerase chain reaction. 2. An antipeptide antibody targeted at a region of the voltage-dependent calcium channel alpha 1D subunit C-terminal to the pore-forming SS1-SS2 loop in domain IV (amino acids 1417-1434) only bound to this exofacial epitope if the DRG neurones and cardiac myocytes were depolarized with 30 mM K+. 3. Incubation of cells under depolarizing conditions for 2-4 h with the antibody resulted in a maximal inhibition of inward current density of 49% (P < 0.005) for DRGs and 30% (P < 0.05) for cardiac myocytes when compared with controls. 4. S-(-)-Bay K 8644 (1 microM) enhanced calcium channel currents in DRGs by 75 +/- 19% (n = 5) in neurones incubated under depolarizing conditions with antibody that had been preabsorbed with its immunizing peptide (100 micrograms ml-1). This was significantly (P < 0.05) larger than the enhancement by S-(-)-Bay K 8644 that was seen with cells incubated under identical conditions but with antibody alone, which was 15 +/- 4% (n = 5). 5. These results demonstrate the presence of calcium channel alpha 1D subunits in rat DRG neurones and guinea-pig cardiac myocytes. They also show that amino acids 1417-1434 of the alpha 1D subunit are only exposed to the extracellular face of the membrane following depolarization and that the binding of an antibody to these amino acids attenuates calcium channel current and reduces the ability of S-(-)-Bay K 8644 to enhance this current, indicating that it is an L-type current that is attenuated.

Amino Acid Sequence↗
PubMed1997

Importance of the different beta subunits in the membrane expression of the alpha1A and alpha2 calcium channel subunits: studies using a depolarization-sensitive alpha1A antibody.

The plasma membrane expression of the rat brain calcium channel subunits alpha1A, alpha2-delta and the beta subunits beta1b, beta2a, beta3b and beta4 was examined by transient expression in COS-7 cells. Neither alpha1A nor alpha2-delta localized to the plasma membrane, either alone or when coexpressed. However, coexpression of alpha1A or alpha2-delta/alpha1A with any of the beta subunits caused alpha1A and alpha2 to be targetted to the plasma membrane. The alpha1A antibody is directed against an exofacial epitope at the mouth of the pore, which is not exposed unless cells are depolarized, both for native alpha1A channels in dorsal root ganglion neurons and for alpha1A expressed with a beta subunit. This subsidiary result provides evidence that either channel opening or inactivation causes a conformational change at the mouth of the pore of alpha1A. Immunostaining for alpha1A was obtained in depolarized non-permeabilized cells, indicating correct orientation in the membrane only when it was coexpressed with a beta subunit. In contrast, beta1b and beta2a were associated with the plasma membrane when expressed alone. However, this is not a prerequisite to target alpha1A to the membrane since beta3 and beta4 alone showed no differential localization, but did direct the translocation of alpha1A to the plasma membrane, suggesting a chaperone role for the beta subunits.

Animals↗
PubMed1995

Voltage-dependent calcium channel beta-subunits in combination with alpha 1 subunits, have a GTPase activating effect to promote the hydrolysis of GTP by G alpha o in rat frontal cortex.

The dihydropyridine-sensitive calcium channel agonist (-)-BayK 8644 was found to produce an enhancement of the intrinsic hydrolysis of GTP by Go in rat frontal cortex membranes. An anti-calcium channel beta-subunit antiserum abolished the (-)-BayK 8644-stimulated hydrolysis of GTP by Go and reduced the dihydropyridine binding capacity of the cortical membranes. A peptide which mimics the beta-subunit binding domain of the calcium channel complex, also attenuated (-)-BayK 8644 activation of GTPase. This study suggests that the calcium channel beta-subunit is the principal component of the channel complex involved in linking dihydropyridine agonist binding to enhanced hydrolysis of GTP by Go. This may be a mechanism by which calcium channels can normally act to limit the duration of a G-protein modulatory signal.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1995

Inhibition of the interaction of G protein G(o) with calcium channels by the calcium channel beta-subunit in rat neurones.

1. The beta-subunit has marked effects on the biophysical and pharmacological properties of voltage-dependent calcium channels. In the present study we examined the ability of the GABAB agonist (-) -baclofen to inhibit calcium channel currents in cultured rat dorsal root ganglion neurones following depletion of beta-subunit immunoreactivity, 108-116 h after microinjection of a beta-subunit antisense oligonucleotide. 2.We observed that, although the calcium channel current was markedly reduced in amplitude following beta-subunit depletion, the residual current (comprising both N- and L-type calcium channel currents) showed an enhanced response to application of (-) -baclofen. Therefore, it is possible that there is normally competition between activated G protein G(o) and the calcium channel beta-subunit for binding to the calcium channel alpha 1-subunit; and this competition shifts in favour of the binding of activated G(o) following depletion of the beta-subunit, resulting in increased inhibition. 3. This hypothesis is supported by evidence that an antibody against the calcium channel beta-subunit completely abolishes stimulation of the GTPase activity of G(o) by the dihydropyridine agonist S-(-) -Bay K 8644 in brain membranes. This stimulation of GTPase is thought to result from an interaction of G(o) alpha-subunit (G alpha o) with its calcium channel effector which may operate as a GTPase-activating protein. 4. These data suggest that the calcium channel beta-subunit when complexed with the beta 1-subunit normally inhibits its association with activated G(o). It may function as a GTPase-activating protein to reduce the ability of activated G(o) to associate with the calcium channel, and thus limit the efficacy of agonists such as (-) -baclofen.

Amino Acid Sequence↗
PubMed1995

Use of site-directed antibodies to probe the topography of the alpha 2 subunit of voltage-gated Ca2+ channels.

Polyclonal antibodies were raised against peptides corresponding to residues 1-15, 469-483 and 933-951 of the rabbit skeletal muscle L-type calcium channel alpha 2/delta primary translation product, for use as topological probes. Immunocytochemical comparison of the abilities of the antibodies to bind to the alpha 2 and delta subunits in intact and detergent-permeabilised rat dorsal root ganglion cells enabled the membrane orientation of these regions to be established. The resultant data indicate that the regions containing residues 1-15 and 469-483 of the alpha 2 subunit, and residues 1-17 of the delta subunit, are exposed on the extracellular surface of the membrane, findings consistent with a model that proposes alpha 2 to be entirely extracellular.

Amino Acid Sequence↗
PubMed1995

Antisense depletion of beta-subunits modulates the biophysical and pharmacological properties of neuronal calcium channels.

1. The role of the voltage-dependent calcium channel (VDCC) beta-subunit has been examined in cultured rat dorsal root ganglion neurones (DRGs). An antipeptide antibody was raised and this recognized proteins corresponding to beta-subunits in a number of preparations. Immunoreactivity for the VDCC beta-subunit in DRGs was concentrated on the internal side of the plasma membrane but was also present in the cytoplasm. 2. A twenty-six-mer antisense oligonucleotide with homology to all published VDCC beta-subunit sequences was microinjected into individual cells, and maximal depletion of VDCC beta-subunit immunoreactivity was observed after 108 h suggesting a half-life for the turnover of the beta-subunit greater than 50 h. No depletion was obtained with nonsense oligonucleotide. 3. The effect of depletion of VDCC beta-subunit immunoreactivity on calcium channel currents in these cells was a reduction in amplitude of the maximum current of about 47%, and a shift in the voltage dependence of current activation of about +7 mV. These effects are the converse of those observed following co-expression of cloned beta- with alpha 1-subunits in oocytes and other expression systems. 4. The ability of the 1,4-dihydropyridine (DHP) agonist Bay K 8644 to enhance calcium channel currents was greatly reduced following depletion of beta-subunit immunoreactivity. This result is in agreement with the finding in several systems that co-expression of the beta-subunit with alpha 1-subunits results in an increased number of DHP binding sites. 5. These results show that calcium channel beta-subunits form part of native neuronal calcium channels and modify their biophysical and pharmacological properties.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1995

Thoracic outlet syndrome after motor vehicle accidents in a Canadian pain clinic population.

OBJECTIVE: To record symptoms and signs, operative findings, and long-term outcome in operated and nonoperated patients with the diagnosis of thoracic outlet syndrome after a motor vehicle accident. DESIGN: Descriptive prospective study. SETTING: Pain clinic population in the Toronto Hospital (Western Division), Toronto, Ontario, Canada. PATIENTS: Thirty-two patients diagnosed as having thoracic outlet syndrome after injuries sustained in a car accident (based on specific symptoms and signs as well as exclusion of other disorders generating brachialgia). INTERVENTIONS: Conservative management versus transaxillary or supraclavicular exploration of the thoracic outlet. OUTCOME MEASURES: Pain/symptom relief. RESULTS: Most patients presented with pain and paresthesiae, but conspicuous discoloration of the symptomatic extremity was seen in 41%. Osseous anomalies were seen in 22% of the patients in simple neck x-rays. Vascular studies were abnormal in 24% of tested patients. During transaxillary first rib resection in 15 patients, 87% were found to have musculotendinous and less often osseous anomalies compromising primarily the lower trunk of the brachial plexus. Long-term follow up demonstrated very good pain relief (based on patients' estimates) only in 47% of the operated patients and 20% of the conservatively treated patients. Reoperation was necessary in six cases through a supraclavicular decompression with only one patient experiencing substantial improvement with long-term follow-up. The difficulties in diagnosis as well as reasons for failures of conservative and surgical management are discussed, and the authors' current protocol for diagnosis and management is presented.

Accidents, Traffic↗
PubMed1994

Effect of different dietary supplemental fats and oils on the tissue fatty acid composition and growth of female broilers.

1. A study was undertaken to examine the effect of supplementation of diets with fats of different chain length and degree of unsaturation on the performance, carcase characteristics and tissue fatty acid composition of broilers. 2. Three hundred and sixty 19-day-old female broilers were fed on diets containing supplemental fat/oil (50 g/kg) in the form of beef tallow (BT) (diet T-T), soyabean oil (SBO) (diet S-S), rapeseed oil (RSO) (diet R-R), marine oil (MO) (diet M-M) or binary mixtures (0.5:0.5 w/w) of these lipids (diets T-S, T-R, T-M, S-R, S-M, and R-M) to 54 d of age. Food intake, live weight, plucked weight and the lipid concentration and fatty acid composition of abdominal fat pad, liver and breast muscle were measured. 3. Food intake, plucked weight and live weight were greatest for diet T-T and lowest for diet R-R. Food conversion ratio was poorest for birds fed on diets containing BT. Lipid concentration in abdominal fat pad was significantly higher in birds fed on diets containing SBO. Liver lipid concentration was significantly reduced by diets containing RSO. 4. Abdominal fat pad fatty acid profile was most readily altered by dietary fatty acids. There was a strong correlation between dietary fatty acid composition and tissue fatty acid composition for all fatty acids except C14:0, C20:0, C20: 2n-6 and C20: 4n-6. The tissue P/S ratio ranged from 0.40 for diet T-T to 1.40 for diet S-S. The n-6/n-3 ratio was significantly increased by the inclusion of SBO and decreased by the inclusion of MO. 5. Liver fatty acid profile was least modified by dietary fatty acids. There was an inverse relationship between liver and dietary C20:4n-6 concentration. Tissue C18:2n-6 and C20:4n-6 were highly correlated, suggesting significant interconversion by delta-6 desaturase in this tissue. The n-6/n-3 ratio was significantly increased by inclusion of SBO and significantly decreased by the inclusion of MO. 6. In breast muscle MO-based diets increased the proportion of n-3 PUFA at the expense of n-6 PUFA. The tissue concentrations of C18:1n-9, C18:2n-6, C20:1n-9, C20:5n-3, C22:5n-3 and C22:6n-3 were strongly correlated with dietary fatty acid composition. Muscle and diet P/S and n-6/n-3 ratios were highly correlated.

Abdomen↗
PubMed1994

Molecular basis for somatostatin action: inhibition of c-fos expression and AP-1 binding.

The mechanisms by which somatostatin exerts its widespread inhibitory actions have been investigated extensively but understood only partially. Recent studies have shown that somatostatin can inhibit gene transcription directly. In view of the critical importance of early response genes in induction of gene expression, we examined whether the action of somatostatin might be mediated by inhibition of early response genes. The products of some of these genes, such as c-fos and c-jun, are known to form a heterodimeric transcription factor complex (AP-1) that binds specifically to the consensus sequence TGAC(G)TCA. Accordingly, we examined the effects of somatostatin on c-fos gene expression and on the binding of the AP-1 complex to its specific DNA element using isolated gastric parietal cells and the GH3 pituitary cell line. In both parietal and GH3 cells, c-fos-specific mRNA was increased by agents known to act via both adenosine-3',5'-cyclic monophosphate and Ca(2+)-dependent signaling mechanisms, and octreotide significantly inhibited this response. Pertussis toxin pretreatment (200 ng/ml) reversed the inhibitory effect of octreotide. AP-1 binding activity, assessed by gel shift assays using a 32P-labeled AP-1 oligonucleotide probe, was stimulated by dibutyryl adenosine 3',5'-cyclic monophosphate and serum and inhibited by octreotide treatment. Our observations support the notion that the universal inhibitory action of somatostatin may be mediated by inhibition of expression of early response genes via a pertussis toxin-sensitive inhibitory pathway. This effect appears to lead to decreased binding of regulatory nuclear proteins to their specific DNA elements resulting, presumably, in diminished gene expression.

Animals↗