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Biomedical subjects

R W Clarke

Publications and source records attributed to R W Clarke.

At least 37 records · Page 2Linked to original sources

Paediatric acute mastoiditis: the Alder Hey experience.

Acute mastoiditis was a common condition in the pre-antibiotic era, but has become rare now with the widespread use of antibiotics. A retrospective study was carried out of patients with acute mastoiditis who were seen at Alder Hey Children's Hospital, Liverpool over a five-year period. Their case records were identified and details of gender, age at presentation, symptoms, signs and clinical management were noted and analysed. Twenty-one patients who presented with acute mastoiditis were identified. The patients' age at presentation ranged from three months to 14 years. Five cases out of 21 (23.8 per cent) were under one year of age. Eighteen cases (85.7 per cent) presented with post-aural swelling while 12 (57.1 per cent) had aural discharge. All patients were treated with intravenous antibiotics and only five patients (23.8 per cent) required surgical intervention in the form of a cortical mastoidectomy with, or without, myringotomy. A significant number of cases can be treated conservatively with intravenous antibiotics. Surgery in the form of cortical mastoidectomy can be reserved for complicated cases and in those in whom conservative treatment has failed.

Acute Disease↗

Differential ability of transition metals to induce pulmonary inflammation.

Transition metals are components of airborne particles and have been implicated in adverse health effects. The relative inflammatory potential of these metals is usually inferred from separate studies that focus on only one or a few individual metals. Comparisons of relative potency among several metals from these separate studies can be difficult. In one comprehensive study, we measured the pulmonary effects of equimolar doses of six metals in soluble form. Our purpose was to compare inflammatory potential and pulmonary toxicity among individual transition metals. Rats received saline, 0.1 or 1.0 micromol/kg of vanadium, nickel, iron(II), copper, manganese, or zinc as sulfates. Bronchoalveolar lavage (BAL) was performed at 0, 4, 16, or 48 h postinstillation. All treatments except V showed increased lactate dehydrogenase activity in BAL fluid; Cu- and Ni-exposed animals had the highest levels. Protein levels in BAL fluid were more than five times higher in Cu-exposed animals compared to other metal treatments at 16 and 48 h. At the 0.1 micromol/kg dose, only Cu induced significant neutrophilia at 16 and 48 h. For the 1.0 micromol/kg dose, all metals tested induced significant neutrophilia, with mean neutrophil numbers for Cu and Mn significantly higher compared to the other metals. At 48 h, neutrophil numbers were still elevated in all metal exposures. Only Mn caused substantial eosinophilia. At the 1.0 micromol/kg dose, only Cu induced macrophage inflammatory protein-2 (MIP-2) mRNA at 4 h. By 48 h, induction of MIP-2 mRNA was observed for all metal exposures except Cu, which subsequently returned to baseline levels. On an equimolar basis, Cu was the most proinflammatory metal, followed by Mn and Ni, while V, Fe(II), and Zn induced similar levels of inflammation. Overall, there were many similarities in the pulmonary responses of the metals we tested. However, we also observed divergent, metal-specific responses. These differential responses suggest that metals induce pulmonary inflammation by differing pathways or combinations of signals.

Actins↗

Endogenous adrenergic control of reflexes evoked by mechanical stimulation of the heel in the decerebrated rabbit.

In decerebrated rabbits, reflexes were evoked in medial gastrocnemius motoneurones by mechanical stimulation of the heel, using four pinch strengths from 183 (innocuous) to 4577 (noxious) mN. The alpha(2)-adrenoceptor antagonist idazoxan (1-156 microg intrathecal (i.th.) significantly increased responses to pinch strengths of 607 mN and above. Subsequent administration of the alpha(1) adrenoceptor selective antagonist prazosin (200 microg i.th.) decreased reflexes to 4577 mN pinches but had no other significant effects. The opioid antagonist (-)-quadazocine (25 microg i.th.) caused no further changes in reflexes. Spinal section at L1 in the presence of this drug combination enhanced gastrocnemius responses to 183 and 607 mN stimuli, had no effect on reflexes to 1866 mN and significantly decreased responses to 4577 mN pinches. These data confirm that reflexes evoked by 'natural' stimulation of heel mechanoreceptors are subject to powerful tonic descending inhibition mediated by alpha(2)-adrenoceptors. For the highest strength stimulus, the results of alpha(2) blockade involved enabling of descending facilitation as well as reduction of descending inhibition.

Adrenergic alpha-Antagonists↗

Adaptive changes in withdrawal reflexes after noxious stimulation at the heel and the toes in the decerebrated rabbit.

In decerebrated rabbits, reflexes evoked by electrical stimulation of the toes in the ankle flexor tibialis anterior were enhanced for > 30 min after application of 20% mustard oil to the base of the toes, whereas responses of the ankle extensor medial gastrocnemius to stimulation of the heel were depressed for > 20 min by the same stimulus. Applied to the heel, mustard oil had inconsistent effects on the flexor reflex but potentiated the extensor response for approximately 1 h. Intrathecal co-administration of naloxone (25 microg) with the selective alpha(2)-adrenoceptor antagonist RX 821002 (200 microg) enhanced both reflexes to more than twice pre-drug values and reduced or abolished all effects of mustard oil. These data confirm that the location of a noxious stimulus is an important determinant of the subsequent adaptive changes in reflexes, and indicate roles for endogenous opioids and noradrenaline in these processes.

Adaptation, Physiological↗

Cannabinoidergic and opioidergic inhibition of spinal reflexes in the decerebrated, spinalized rabbit.

The present experiments were designed to investigate the role(s) of cannabinoid receptors in modulating transmission in the sural-medial gastrocnemius withdrawal reflex of the decerebrated, spinalized rabbit and how, if present, cannabinoid-mediated control might interact with opioid-mediated inhibitions known to impinge on this reflex pathway. The selective CB(1) receptor antagonist SR 141716A enhanced reflexes by a factor of two after a cumulative dose of 100 nmol kg(-1) i.v., but had no effect on the endogenous opioid-mediated inhibition generated by repetitive electrical stimulation of the common peroneal nerve, or on the suppression of reflexes caused by i.v. administration of the synthetic opioid fentanyl. Given at a dose of 10 nmol kg(-1) i.v., the potent, CB(1)--CB(2) cannabinoid receptor agonist HU 210 inhibited medial gastrocnemius reflexes to approximately 30% of controls and significantly decreased both heart rate and blood pressure, but did not alter the inhibition of reflexes resulting from common peroneal nerve stimulation or i.v. fentanyl. The effects of HU 210 were reversed by SR 141716A. HU 210 was just as effective in inhibiting reflexes in the presence of the opioid antagonist naloxone (5 micromol kg(-1)) as it was in untreated animals. The data show that cannabinoids, acting through CB(1) receptors, are inhibitory in rabbit spinal cord and that there appears to be some endogenous cannabinoid tone under the conditions of the present experiments. The evidence of this study is that the inhibitory effects of opioids and cannabinoids in rabbit spinal cord are completely independent of each other, and are additive rather than synergistic.

Animals↗

Opioid and GABA receptors involved in mediation and modulation of tonic and stimulus-evoked inhibition of a spinal reflex in the decerebrated and spinalized rabbit.

The purposes of this study were to investigate: (i) the identity of the opioid peptide(s) mediating tonic and stimulus-evoked inhibition of the sural-medial gastrocnemius reflex of the decerebrated, spinalized rabbit and (ii) the modulation of these processes by endogenous GABA. The selective delta receptor antagonist naltrindole (100 nmol kg(-1) i.v.), the GABA(A) blocker bicuculline (300 nmol intrathecal, i.th.), and the GABA(B) antagonist CGP 35348 (1 micromol i.th.) increased gastrocnemius reflexes to 150-160% of pre-drug values, whereas a sub-maximal dose of naloxone (30 nmol kg(-1) i.v.) augmented reflexes to >500% of controls. Kelatorphan, an inhibitor of enkephalin metabolism (2 micromol i.th.), depressed gastrocnemius responses by 50% and potentiated the inhibitory effects of methionine enkephalin. Repetitive electrical stimulation of the superficial or common peroneal nerves inhibited reflexes for 15-20 min. This effect was significantly reduced by naltrindole and CGP 35348. It was not reduced by a low dose (30 nmol kg(-1) i.v.) of naloxone or by bicuculline. When naloxone and naltrindole were combined at 30 nmol kg(-1) each, stimulus-evoked inhibition was blocked. Given after bicuculline, naloxone at 100 nmol kg(-1) i.v. abolished peroneal-evoked inhibition, but a dose of 300 nmol kg(-1) was required to produce the same effect after CGP 35348. Kelatorphan augmented the depth and duration of inhibition evoked by peroneal nerve stimulation. These data are consistent with the involvement of enkephalin-like peptides in tonic and stimulus-evoked inhibition of the sural-gastrocnemius reflex. Tonic inhibition in rabbit spinal cord is dominated by opioids acting through mu receptors, whereas co-activation of delta, mu and GABA(B) receptors mediates stimulus-evoked inhibition. It is possible that GABA(B) receptors inhibit the release of spinal opioids while simultaneously supporting their actions at post-synaptic targets.

Animals↗

The spatial organization of central sensitization of hind limb flexor reflexes in the decerebrated, spinalized rabbit.

This study was designed to investigate the relationship between the location of a noxious stimulus and the magnitude and duration of the plastic effects induced by that stimulus in withdrawal reflexes acting about the knee and the ankle, in rabbits. Reflexes were evoked in the nerves to the anterior tibial and semitendinosus flexor muscles by electrical stimulation at the toes. Repetitive, high intensity electrical stimulation of nerve trunks (sural, medial gastrocnemius, superficial peroneal, tibial, 100 pulses, 20 V, 1 ms at 0.5 Hz) was generally found to be a poor method for inducing central sensitization in these flexor reflexes. 'Natural' noxious stimulation induced more reliable enhancement of both reflexes. Mechanical (clamp) or chemical (mustard oil) stimulation of the heel induced prolonged (median duration >30 min) increases in reflexes to both muscles. Mechanical (clamp and superficial pinch) or chemical (mustard oil) stimulation of the toes, and injection of bradykinin into the gastrocnemius muscles or into the soft tissues of the sole of the foot, also led to enhancement of both reflexes, with the median duration of potentiation between 7 and 30 min. The effects obtained from deep tissue stimulation were generally weaker than those obtained after stimulation of superficial structures. These data show that there were no major differences in the effects obtained from the heel vs the toes, or between the two reflexes. It appears that the spatial organization of the spinal mechanisms underlying central sensitization of flexor withdrawal reflexes is rather more crudely drawn than that pertaining to the reflexes themselves. Furthermore, the data indicate that in the present preparation, afferents from deep tissues are no more effective in generating central sensitization than those from superficial structures.

Afferent Pathways↗

Strain-related differences of nonspecific respiratory defense mechanisms in rats using a pulmonary infectivity model.

A number of animal studies have assessed pulmonary host defense mechanisms by inoculating the lungs with the bacterial agent, Listeria monocytogenes. Most studies use only a single strain of the animal to be tested; however, strain-related differences in responsiveness to pulmonary toxicants have been well documented. It was the goal of this current investigation to measure the pulmonary defense responses of two different strains of rats in a lung infectivity model. Fischer 344 (F344) and Sprague-Dawley (SD) rats were instilled intratracheally with 5 x 10(3) or 5 x 10(5) L. monocytogenes, and the effect on mortality, lung injury and inflammation, pulmonary bacterial clearance, and alveolar macrophage (AM) function was determined at 3, 5, and 7 days after bacteria treatment. Pulmonary inoculation with the higher (5 x 10(5) L. monocytogenes) dose proved to be highly pneumotoxic to the F344 rats as evidenced by an increase in mortality and more severe lung injury and inflammation when compared with the SD rats. After intratracheal instillation with the lower (5 x 10(3) L. monocytogenes) dose, pulmonary bacterial clearance was slowed and an increase in pulmonary responsiveness was observed for the F344 rats as compared to the SD rats. Specifically, the total number of neutrophils recovered from the lungs and tumor necrosis factor-alpha secreted by AMs were elevated for the F344 group throughout the 7 days, while cellular chemiluminescence, an index of reactive oxygen species production, and lung albumin and lactate dehydrogenase, indicators of injury, were increased at 3 and 5 days after bacterial instillation. This study demonstrated that respiratory defense function was compromised in F344 rats as evidenced by elevated mortality, slowed pulmonary bacterial clearance, and altered AM function. F344 rats may then represent a sensitive model for the examination of respiratory defense mechanisms after bacterial challenge.

Albumins↗

Effect of age on respiratory defense mechanisms: pulmonary bacterial clearance in Fischer 344 rats after intratracheal instillation of Listeria monocytogenes.

STUDY OBJECTIVES: To examine the lung defense mechanisms of both young and aged rats before and after pulmonary challenge with a bacterial pathogen. DESIGN: Male Fischer 344 rats, either 2.5 months or 20 months of age, were intratracheally inoculated with 5 x 10(3), 5 x 10(4), or 5 x 10(5) Listeria monocytogenes, and the effects on mortality, lung inflammation, pulmonary bacterial clearance, alveolar macrophage (AM) function, and T-lymphocyte characterization were determined. MEASUREMENTS AND RESULTS: In noninfected control animals, the older rats had lower numbers of AMs on lavage and a lower percentage of total T, CD4+, and CD8+ cells. No difference was observed between noninfected young and old rats in AM function, assessing both chemiluminescence and nitric oxide (NO) production. After bacterial challenge, aged rats exhibited an increase in mortality, pulmonary infection, and edema, and lung lesions, which were more extensive than those observed in the younger rats. Interestingly, AM chemiluminescence was enhanced, while AM NO, a highly important antibacterial defense product, was abrogated in the aged rats as compared to the young rats. CONCLUSIONS: This study demonstrated that advanced age is associated with alterations in lung defense mechanisms and increased susceptibility to pulmonary bacterial infection marked by elevated mortality, slowed pulmonary bacterial clearance, and altered AM function, specifically a decrease in NO production. These observations are indicative of reduced pulmonary defense function in an older population of rats.

Aging↗

Imidazoline I(2)-receptors and spinal reflexes in the decerebrated rabbit.

Idazoxan potentiates spinal reflexes in the decerebrated rabbit, an effect that has been attributed to antagonism of tonic noradrenergic inhibition. It is now known that this drug has a higher affinity for I(2)-imidazoline receptors than alpha(2)-receptors. The roles of I(2)-receptors in modulating transmission in spinal reflex pathways have been investigated using the selective ligands RX 821029 and RS-45041-190, and, as I(2)-receptors are closely associated with monoamine oxidase (MAO), the MAO inhibitors pargyline and clorgyline. In decerebrated rabbits with an intact spinal cord, intrathecal doses of 5-365 microg (cumulative) of the I(2)-ligands augmented, to 150-180% of pre-drug levels, the reflex responses of medial gastrocnemius motoneurones to electrical stimulation of the sural nerve, and significantly increased mean arterial blood pressure by approximately 10 mmHg over pre-drug values. Neither MAO inhibitor had significant effects on reflexes, but the highest dose of the MAO-A selective agent clorgyline (365 microg cumulative) caused a significant rise in blood pressure of 6 mmHg. Neither the I(2)-ligands nor the MAO inhibitors prevented the further enhancement of reflexes or blood pressure by subsequent administration of the selective alpha(2)-adrenoceptor antagonist RX 821002. In decerebrated, spinalized rabbits, intrathecal RS-45041-190 (60 microg, single dose) increased spinal reflex responses to 109% of pre-drug values, a significantly smaller effect than that seen in non-spinal preparations, and had no effect on blood pressure. These data show that imidazoline I(2)-receptors can influence somatic and autonomic motor outflows. These effects should be taken into account when interpreting the spinal effects of imidazoline-based adrenoceptor-active drugs.

Adrenergic alpha-Antagonists↗

The involvement of tachykinin NK2 and NK3 receptors in central sensitization of a spinal withdrawal reflex in the decerebrated, spinalized rabbit.

Repetitive electrical stimulation of the sural nerve at a strength sufficient to excite C-fibres results in prolonged (> 20 min) increases in the reflex responses of gastrocnemius motoneurones evoked by stimulation of myelinated axons in the sural nerve. We have tested the susceptibility of this effect to blockade of tachykinin NK2 and NK1 receptors. In the control state, iterative stimulation of sural nerve C fibres increased gastrocnemius reflexes to a peak of between 200 and 400% of pre-stimulus levels, an effect that recovered to baseline values over 23-30 min. Pre-treatment with selective antagonists for NK2 (SR 48968, 1 mg/kg i.v.) or NK3 (SR 142801, 0.1 and 1 mg/kg i.v.) receptors failed to alter the peak facilitation resulting from sural nerve stimulation. However, both drugs significantly reduced the duration of enhancement of reflexes, to 18 min after SR 48968 and to 5 min after SR 142801. The inactive enantiomers of these compounds (SR 48965 and SR 142806, both 1 mg/kg i.v.) did not reduce facilitation of reflexes after sural nerve stimulation. We conclude that activation of tachykinin NK3 receptors is a major factor in maintaining central sensitization of the sural-gastrocnemius reflex after electrical stimulation of sural nerve C-fibres. The effects of SR 48968 were surprisingly weak and may have resulted from an effect of this compound at NK3 receptors.

Animals↗

The role of 5-HT(1A)-receptors in fentanyl-induced bulbospinal inhibition of a spinal withdrawal reflex in the rabbit.

The sural to gastrocnemius withdrawal reflex is inhibited after injection of the OP(3) (micro)-receptor-selective opioid fentanyl into the fourth ventricle of decerebrated rabbits. This effect is abolished by complete section of the spinal cord but not by the selective alpha(2)-adrenoceptor antagonist RX 821002 (Clarke RW, Parry-Baggott C, Houghton AK, Ogilvie J. The involvement of bulbo-spinal pathways in fentanyl-induced inhibition of spinal withdrawal reflexes in the decerebrated rabbit. Pain 1998;78:197-207). We have now investigated the role of 5-HT(1A) receptors in mediating the descending inhibition activated by intraventricular fentanyl. In the control state, intraventricular fentanyl (3-30 microgram/kg) inhibited gastrocnemius reflex responses to a median of 34% of pre-drug levels. After intrathecal administration of the selective 5-HT(1A) receptor antagonist WAY-100635 (100 microgram), fentanyl reduced reflex responses to 83% of pre-fentanyl values, significantly less inhibition than in the control state. In a separate group of experiments, intravenous fentanyl (0.3-30 microgram/kg) depressed the sural-gastrocnemius reflex to 17% of pre-drug controls. This inhibition was not affected by intrathecal WAY-100635 (100 microgram), but combined administration of the 5-HT(1A) antagonist with RX 821002 (100 microgram) significantly reduced the effectiveness of i.v. fentanyl. After the highest dose reflexes were 37% of pre-fentanyl levels. These data show that the bulbospinal inhibition activated by fentanyl is mediated, at least in part, by activation of spinal 5-HT(1A) receptors. That blockade of these receptors failed to influence the inhibition induced by i.v. fentanyl might be taken to mean that the brain-stem action of fentanyl does not contribute significantly to the systemic actions of this opioid. A more probable explanation is that, in the preparation used in the present study, the bulbospinal and direct spinal actions of fentanyl occlude each other to produce an overall inhibition that is less than the sum of the two effects.

Adrenergic alpha-Antagonists↗

Effects of a selective 5-HT(1B/1D) receptor agonist on spinal and trigeminal reflexes in the anaesthetized rabbit.

The effects of the 5-HT(1B/1D) receptor agonist L-741,604 on a trigeminally-mediated (jaw depressor) reflex and a spinally-mediated (flexion withdrawal) reflex have been compared between spinalized and intact, anaesthetized rabbits. L-741,604 depressed the jaw depressor reflex dose-dependently in all animals, to a median of 5% (inter-quartile range, IQR, 3 - 28%, n=18) of pre-drug levels after a cumulative dose of 3.1 micromol kg(-1) i.v. This effect was reversed by the 5-HT(1B/1D) antagonist GR 127,935 (1 - 2 micromol kg(-1) i.v.). The flexion withdrawal reflex was depressed by L-741, 604 in non-spinalized animals, to a median of 22% (IQR 10 - 36%, n=10) of pre-drug levels after the highest dose, an action that was reversed by GR 127,935. In spinalized rabbits, L-741,604 up to 0.3 micromol kg(-1) i.v. cumulative increased the flexion reflex to a median of 189% (IQR 169 - 198%, n=8) of pre-drug controls. With higher doses the reflex decreased, so that after 3.1 micromol kg(-1) it was 75% (IQR 55 - 96%) of pre-drug levels. Subsequent GR 127,935 increased reflexes to a median of 180% (IQR 136 - 219%) of controls. L-741,604 increased arterial blood pressure and decreased heart rate in both preparations, effects that were reversed by GR 127,935. Thus, when the spinal cord was intact L-741,604 inhibited spinal and trigeminal reflexes in the same way. Although spinalization enabled a non-5-HT(1B/1D)-mediated excitatory effect of L-741,604 on spinal reflexes, there was a clear inhibitory effect of the drug at high doses. These data suggest that L-741,604 inhibits spinal reflexes by increasing descending inhibition and by a direct action in the cord. The same processes could apply to inhibition of trigeminally-mediated events.

Animals↗

Inhaled particle-bound sulfate: effects on pulmonary inflammatory responses and alveolar macrophage function.

Acid sulfate-coated solid particles are a significant environmental hazard produced primarily by the combustion of fossil fuels. We have previously described a system for the nascent generation of carbonaceous particles surface coated with approximately 140 microg/m(3) acid sulfate [cpSO(4)(2-); 10 mg/m(3) carbon black (CB) and 10 ppm sulfur dioxide (SO(2)) at 85% relative humidity (RH)]. The effects of inhaled cpSO(4)(2-) on pulmonary host defenses are assessed in the present work. Mice were acutely exposed (4 h) to either 10 mg/m(3) CB, 10 ppm SO(2), or their combination at 10% or 85% RH in a nose-only inhalation chamber. No evidence of an inflammatory response was found following any of the exposures as assessed by total cell counts and differential cell counts from bronchoalveolar lavage fluid. However, alveolar macrophage Fc receptor-mediated phagocytosis decreased only following exposure to 140 microg cpSO(4)(2-), significant suppression occurred after 24 h, maximal suppression occurred at 3 days postexposure, and recovery to preexposure levels required 7-14 days. Intrapulmonary bactericidal activity (IBA) was also suppressed only after exposure to 140 microg cpSO(4)(2-); suppression was maximal at 1 day postexposure and recovered by day 7. To assess the effects of lower cpSO(4)(2-) concentrations, mice were repeatedly exposed to 1 mg/m(3) CB and 1 ppm SO(2) at 85% RH ( approximately 20 microg/m(3) cpSO(4)(2-) for 4 h/day) for up to 6 days. A significant decrement in IBA was observed following 5 and 6 days of exposure. These studies indicated that acute or repeated exposure to cpSO(4)(2-) could alter pulmonary host defense mechanisms.

Administration, Inhalation↗

Interstrain variation in murine susceptibility to inhaled acid-coated particles.

Epidemiologic studies have demonstrated a positive correlation between concentration of acid aerosol and increased morbidity and mortality in many urban environments. To determine whether genetic background is an important risk factor for susceptibility to the toxic effects of inhaled particles, we studied the interstrain (genetic) and intrastrain (environmental) variance of lung responses to acid-coated particle (ACP) aerosol in nine strains of inbred mice. A flow-past nose-only inhalation system was used to expose mice to ACPs produced by the cogeneration of a carbon black aerosol-sulfur dioxide (SO(2)) mixture at high humidity. Three days after a single 4-h exposure to ACPs or filtered air, mice underwent bronchoalveolar lavage, and cell differentials and total protein were determined as indexes of inflammation and epithelial permeability, respectively. To determine the effect of ACPs on alveolar macrophage (AM) function, lavaged AMs were isolated from exposed animals and Fc receptor-mediated phagocytosis was evaluated. Compared with air-exposed animals, there was a slight but significant exposure effect of ACPs on the mean number of lavageable polymorphonuclear leukocytes in C3H/HeJ and C3H/HeOuJ mice. ACP exposure also caused a significant decrease in AM phagocytosis. Relative to respective air-exposed animals, Fc receptor-mediated phagocytosis was suppressed in eight of nine strains. The order of strain-specific effect of ACPs on phagocytosis was C57BL/6J > 129/J > SJL/J > BALB/cJ > C3H/HeOuJ > A/J > SWR/J > AKR/J. There was no effect of ACP exposure on AM phagocytosis in C3H/HeJ mice. The significant interstrain variation in AM response to particle challenge indicates that genetic background has an important role in susceptibility. The effects of ACPs on AM function, inflammation, and epithelial hyperpermeability were not correlated (i.e., no cosegregation). This model may have important implications concerning interindividual variation in particle-induced compromise of host defense.

Acids↗

Genetic linkage analysis of susceptibility to particle exposure in mice.

Particle-induced increases in respiratory morbidity and mortality have been observed worldwide in industrialized cities but the toxicologic mechanisms have not been elucidated. It is hypothesized that subpopulations including the elderly and individuals with cardiopulmonary disease are particularly at risk to the effects of exposure. Genetic background is another important host factor that may contribute to interindividual responsivity to particulate exposure. This study was designed to identify susceptibility loci for alveolar macrophage (AM) immune dysfunction induced by inhalation of sulfate-associated carbon particles in susceptible C57BL/6J and resistant C3H/ HeJ inbred mice. AMs were chosen for study because they represent an important component of host defense, and compromised host defense has been hypothesized to be an important factor in particle-induced respiratory morbidity. The quantitative phenotype for these studies was Fc receptor-mediated phagocytic function, an index of AM integrity. Analyses of macrophage dysfunction phenotypes of segregant and nonsegregant populations derived from these two strains indicate that two unlinked genes control susceptibility. A genome-wide linkage analysis of an intercross (F(2)) cohort identified significant and suggestive quantitative trait loci (QTLs) on chromosomes 17 and 11, respectively. Candidate susceptibility genes were identified for mice and humans by comparative mapping. Importantly, both QTLs overlap previously identified QTLs for susceptibility to another common pollutant, ozone. This is the first demonstration that genetic background is an important determinant of responsiveness to particle-induced immune dysfunction, and it has important implications for understanding the epidemiologic associations between particulates and morbidity and mortality.

Air Pollution↗

Inhaled concentrated ambient particles are associated with hematologic and bronchoalveolar lavage changes in canines.

Pulmonary inflammatory and hematologic responses of canines were studied after exposure to concentrated ambient particles (CAPs) using the Harvard ambient particle concentrator (HAPC). For pulmonary inflammatory studies, normal dogs were exposed in pairs to either CAPs or filtered air (paired studies) for 6 hr/day on 3 consecutive days. For hematologic studies, dogs were exposed for 6 hr/day for 3 consecutive days with one receiving CAPs while the other was simultaneously exposed to filtered air; crossover of exposure took place the following week (crossover studies). Physicochemical characterization of CAPs exposure samples included measurements of particle mass, size distribution, and composition. No statistical differences in biologic responses were found when all CAPs and all sham exposures were compared. However, the variability in biologic response was considerably higher with CAPs exposure. Subsequent exploratory graphical analyses and mixed linear regression analyses suggested associations between CAPs constituents and biologic responses. Factor analysis was applied to the compositional data from paired and crossover experiments to determine elements consistently associated with each other in CAPs samples. In paired experiments, four factors were identified; in crossover studies, a total of six factors were observed. Bronchoalveolar lavage (BAL) and hematologic data were regressed on the factor scores. Increased BAL neutrophil percentage, total peripheral white blood cell (WBC) counts, circulating neutrophils, and circulating lymphocytes were associated with increases in the aluminum/silicon factor. Increased circulating neutrophils and increased BAL macrophages were associated with the vanadium/nickel factor. Increased BAL neutrophils were associated with the bromine/lead factor when only the compositional data from the third day of CAPs exposure were used. Significant decreases in red blood cell counts and hemoglobin levels were correlated with the sulfur factor. BAL or hematologic parameters were not associated with increases in total CAPs mass concentration. These data suggest that CAPs inhalation is associated with subtle alterations in pulmonary and systemic cell profiles, and specific components of CAPs may be responsible for these biologic responses.

Air Pollutants↗

Mechanisms of morbidity and mortality from exposure to ambient air particles.

The studies reported here assessed pathophysiologic mechanisms that result from exposure to concentrated ambient particles (CAPs) in animals with and without cardiopulmonary compromise. These studies were carried out to determine the biologic plausibility of epidemiologic observations of increases in particulate air pollution associated with increases in human morbidity and mortality. Dogs were exposed two at a time to CAPs or filtered air via tracheostomy for six hours per day on three consecutive days. The electrocardiogram (ECG) and breathing pattern were recorded continuously, and indicators of inflammation were also assessed. In one experimental design, normal dogs were exposed in pairs to CAPs and subsequently to filtered air or to filtered air and subsequently CAPs (the double CAPs/double sham design). Comparisons were made between the CAPs measurements and each dog's own sham responses. In another design, one dog was exposed to CAPs while the chambermate received a sham exposure; these experiments were followed by crossover of the protocol the subsequent week (the crossover design). Comparisons were made between the CAPs exposure and both the chambermate's sham and each dog's own sham responses. The crossover experiments were conducted in normal animals and in animals who had undergone balloon occlusion of the left anterior descending (LAD) coronary artery to induce myocardial compromise. The effects of CAPs in animals with induced chronic bronchitis were part of the original specific aims; because these studies were not fully pursued, the results are presented only in Appendix A. In normal dogs, analyses of all double CAPs and crossover studies revealed low frequency (LF) and high frequency (HF) powers for heart rate variability (HRV) that were significantly higher for CAPs exposure compared to sham exposure. Variation in day-to-day exposure concentrations, aerosol composition, and pathophysiologic responses were also found. The crossover design, continuous measures of aerosol mass, and biologic responses were incorporated in the development of a statistical model that allowed isolation of changes associated with CAPs from changes due to animal variations. Comparison of individual exposures with this model revealed a range from no response in any measured parameter to statistically significant changes in cardiac autonomic balance, pulmonary air flow, and breathing pattern. On days in which dogs showed statistically significant changes in responses, the findings were consistent in both cardiac and respiratory parameters. Days associated with significant increases in LF and HF HRV, LF/ HF HRV ratio, and heart rate standard deviation (HR SD) were also associated with decreases in average heart rate. These same days had decreases in respiratory frequency, tidal volume, minute volume, and peak flows with corresponding increases in respiratory cycle times and enhanced pause (Pauenh), a measure of bronchoconstriction. These cardiac and respiratory changes suggest an effect mediated via both the sympathetic nervous system and the vagus nerve. Alternatively, days associated with increased heart rate had decreases in the HR SD; decreases or no change in HF and LF HRV; increases in respiratory flows and volumes; and decreases in breathing cycle times, all suggesting only sympathetic nervous system mediation. When all data from the crossover design experiments were assessed with this model, the heart rate and respiratory rate were significantly decreased in relation to both cumulative and actual exposure and the LF HRV, LF/HF HRV ratio, HR SD, and all other respiratory parameters were significantly increased (p < 0.0001 for all). When cardiac data were grouped by days in which the air mass trajectory came from the north or northwest (versus west, south, east, or northeast), significant increases in HR SD and HF HRV and significant decreases in average heart rate were associated with the northwest trajectory. (ABSTRACT TRUNCATED)

Air Pollutants↗