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Biomedical subjects

M Ennis

Publications and source records attributed to M Ennis.

At least 91 records · Page 5Linked to original sources
PubMed1998

In vitro effects of the pyrethroid S-bioallethrin on lymphocytes and basophils from atopic and nonatopic subjects.

Synthetic pyrethroids are increasingly used as insecticides and marketed as having relatively low human toxicity. The aim of this study was to examine the in vitro effects of the synthetic pyrethroid S-bioallethrin on human blood lymphocytes and basophils in atopic individuals and nonatopic control subjects. S-bioallethrin caused inhibition of lymphocyte proliferation after a 72-h culture period in a concentration-dependent manner. The inhibition of the lymphocyte proliferation by S-bioallethrin at the concentration 6.5 microM correlated well with the total serum IgE values (r = -0.89, P < 0.001). Samples from atopic subjects were more sensitive to this inhibition than those from nonatopic volunteers. The regulatory interleukin-4/interferon-gamma (JL-4/IFN-gamma) balance showed a significant difference between atopic and nonatopic subjects after a short-term culture period (24 h) in the presence of the same concentration range of S-bioallethrin (P < 0.001). Additionally, IFN-gamma secretion was consistently lower in cells from the atopic donors. Furthermore, S-bioallethrin induced histamine release from human basophils in a concentration-dependent manner. Although the effect was small compared to histamine liberators such as N-formyl-Met-Leu-Phe and anti-IgE, the response to S-bioallethrin was significantly different in atopic donors from nonatopic (P = 0.0431). These findings are the first demonstration of the immunotoxicologic properties of the synthetic pyrethroid S-bioallethrin by this combined in vitro approach with human lymphocytes and basophils. Further studies will investigate the responses of lymphocytes from patients who are sensitive to these agents.

Adolescent↗
PubMed1998

Neurokinin A is the predominant tachykinin in human bronchoalveolar lavage fluid in normal and asthmatic subjects.

BACKGROUND: Multiple sensory neuropeptides are present in human airways and may contribute to diseases such as asthma. This study quantified and characterised substance P (SP), neurokinin A (NKA), and calcitonin gene related peptide (CGRP) immunoreactivity in bronchoalveolar lavage fluid in asthmatic and normal subjects. METHODS: Using specific radioimmunoassay (RIA), SP, NKA and CGRP were measured in bronchoalveolar lavage fluid from asthmatic subjects (n = 5), normal subjects (n = 5), atopic non-asthmatic subjects (n = 6), and asthmatic subjects four hours after allergen challenge (n = 12). Peptide immunoreactivity was characterised using high performance liquid chromatography (HPLC) and RIA. RESULTS: No SP or CGRP immunoreactivity was detected in any of the fractions from samples after extraction, HPLC, and RIA. Non-specific binding resulted in spurious SP immunoreactivity being detected in bronchoalveolar lavage fluid when no extraction process was employed. NKA was detected in significant amounts in asthmatic (median 550, range 425-625 pg/ml) and normal subjects (median 725, range 350-1425 pg/ml). The level of NKA was significantly higher in the asthmatic subjects after allergen challenge (median 750, range 350-1250 pg/ml) than in unchallenged asthmatic subjects (median 600, range 425-600 pg/ml, p < 0.01). CONCLUSIONS: Extraction and characterisation of peptides from bronchoalveolar lavage fluid must be performed to ensure that the measured immunoreactivity represents target peptide. NKA is present in bronchoalveolar lavage fluid in high concentrations and is the predominant tachykinin. The concentrations of NKA are similar in normal subjects and subjects with mild asthma.

Adolescent↗
PubMed1998

Evaluation and outcome of patients with chronic non-productive cough using a comprehensive diagnostic protocol.

BACKGROUND: Asthma, post-nasal drip syndrome (PNDS), and gastro-oesophageal reflux (GOR) account for many cases of chronic non-productive cough (CNPC). Each may simultaneously contribute to cough even when clinically silent, and failure to recognise their contribution may lead to unsuccessful treatment. METHODS: Patients (all lifetime non-smokers with normal chest radiographs and spirometric measurements) referred with CNPC persisting for more than three weeks as their sole respiratory symptom underwent histamine challenge, home peak flow measurements, ear, nose and throat (ENT) examination, sinus CT scanning, and 24 hour oesophageal pH monitoring. Treatment was prescribed on the basis of diagnoses informed by investigation results. RESULTS: Forty three patients (29 women) of mean age 47.5 years (range 18-77) and mean cough duration 67 months (range 2-240) were evaluated. On the basis of a successful response to treatment, a cause for the cough was identified in 35 patients (82%) as follows: cough variant asthma (CVA) (10 cases), PNDS (9 cases), GOR (8 cases), and dual aetiologies (8 cases). Histamine challenge correctly predicted CVA in 15 of 17 (88%) positive tests. ENT examination and sinus CT scans each had low positive predictive values for PNDS (10 of 16 (63%) and 12 of 18 (67%) positive cases, respectively), suggesting that upper airways disease frequently co-exists but does not always contribute to cough. When negative, histamine challenge and 24 hour oesophageal pH monitoring effectively ruled out CVA and GOR, respectively, as a cause for cough. CONCLUSION: This comprehensive approach aids the accurate direction of treatment and, while CVA, PNDS and GOR remain the most important causes of CNPC to consider, a group with no identifiable aetiology remains.

Adolescent↗
PubMed1997

Soman-induced seizures rapidly activate astrocytes and microglia in discrete brain regions.

Neurons in the piriform cortex and the pontine nucleus locus coeruleus express elevated levels of the immediate early gene protein product, Fos, within 30-45 minutes of a seizurogenic dose of the anticholinesterase, soman (Zimmer et al., [1997] J. Comp. Neurol. 378:468-481). By 24 hours following soman injection, there is marked neuropathology in the piriform cortex. These findings suggest selective, regional vulnerability in response to the seizurogenic actions of soman. In the present study, we determined that soman-induced seizures also cause selective, rapid activation of astrocytes and microglia in the piriform cortex and other brain regions. Animals were killed at different intervals between 1 hour and 24 hours after a convulsive dose of soman. Brain sections were processed for immunocytochemical detection of astrocytes with antibodies against glial fibrillary acidic protein, and microglia and macrophages with antibodies against the complement receptor 3 protein, OX-42. The results demonstrate that following soman administration: (1) there is a rapid increase in glial fibrillary acidic protein staining in astrocytes of the piriform cortex (1 hour); (ii) reactive astrocytes are specifically restricted to layer II and the superficial boundaries of layer III of the piriform cortex. These are the same layers in which neurons express Fos within 30-45 minutes following soman administration; (3) between 1 and 4 hours, resting (ramified) microglia in the piriform cortex and the hippocampus alter their morphology to resemble active microglia. From 4-8 hours, active microglia undergo morphological changes characteristic of reactive microglia that resemble macrophages. Taken together, these observations indicate that astrocytes and microglia in brain regions susceptible to soman become rapidly "reactive" in response to seizures. The highly specific anatomical codistribution of reactive glia and Fos-expressing neurons suggests that intensely active neurons provide local signals that trigger reactive changes in neighboring glia.

Animals↗
PubMed1997

Anatomical localization and time course of Fos expression following soman-induced seizures.

Soman (pinacolymethylphosphonofluoridate), a highly potent, irreversible inhibitor of cholinesterase, causes intense convulsions, neuropathology and, ultimately, death. There is evidence that certain brain structures are selectively vulnerable to the pathological consequences of soman-induced seizures. A working hypothesis is that central nervous system (CNS) structures with the earliest and most severe signs of neuropathology may be key sites for the initiation of the seizures. Fos, the immediate-early gene product, increases rapidly in several animal seizure models. Thus, we reasoned that the earliest brain regions to express Fos might be involved in the initiation and maintenance of soman-induced convulsions. To assess this, rats were injected with a single, convulsive dose of soman (77.7 micrograms/kg, i.m.). The animals were euthanized and processed for immunocytochemical analysis at several time points. Robust Fos expression was seen in layer II of the piriform cortex and the noradrenergic nucleus locus coeruleus within 30-45 minutes. One hour following soman injection, staining was more intense in the piriform cortex layer II and in the locus coeruleus. In addition, Fos was evident in the piriform cortex layer III, the entorhinal cortex, the endopiriform nucleus, the olfactory tubercle, the anterior olfactory nucleus and the main olfactory bulb. By 2 hours, Fos staining was present throughout the cerebral cortex, thalamus, caudate-putamen and the hippocampus. At 8 hours and beyond, Fos expression returned to control levels throughout the CNS except for the piriform cortex and the locus coeruleus which still had robust labeling. By 24 hours, neuropathology was evident throughout the rostral-caudal extent of layer II of the piriform cortex. The rapid induction of Fos in the piriform cortex and the locus coeruleus, taken together with previous anatomical, eletrophysiological and neurochemical studies, suggests that prolonged, excessive exposure to synaptically released acetylcholine and norepinephrine triggers the production of soman-induced seizures initially in the piriform cortex and subsequently in other cortical and subcortical structures.

Animals↗
PubMed1997

Histamine release from human bronchoalveolar lavage mast cells by neurokinin A and bradykinin.

OBJECTIVE AND DESIGN: This study examined whether bradykinin and neurokinin A activate human pulmonary mast cells retrieved by bronchoalveolar lavage (BAL). SUBJECTS: BAL samples were obtained at routine bronchoscopy from 14 unpreselected patients. METHODS: Histamine release experiments were performed using substance P, neurokinin A, bradykinin (peptides 25 and 50 microM), compound 48/80 (0.75-10 micrograms/ml) and A23187 (1 microM). Statistical analyses were performed using the paired Student's t-test and Pearson's linear correlation coefficient. RESULTS: Compound 48/80 induced release was significantly lower than that induced by the other secretagogues (p < 0.05). Neurokinin A and bradykinin induced release correlated significantly with substance P induced release (p < 0.01), suggesting similar mechanisms of action. No correlations were observed between neurokinin A or bradykinin-induced release and the non-peptide stimuli studied. CONCLUSIONS: The mechanism of neurokinin A- and bradykinin-induced bronchoconstriction is not yet clear but our data suggest an indirect effect mediated by mast cell degranulation.

Bradykinin↗
PubMed1997

Glomerular synaptic responses to olfactory nerve input in rat olfactory bulb slices.

In olfactory bulb slices from young rats, the field potential evoked in the glomerular layer by stimulation in the olfactory nerve layer consisted of two negative components: an early component (N1) which was blocked by bath application of the kainate/amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM), and a late, prolonged component (N2; duration > or = 350 msec) which was unaffected by CNQX, was enhanced by reduction of Mg2+ in the medium, and was blocked by the N-methyl-D-aspartate receptor antagonist DL-2-amino-5-phosphonovalerate (50 microM). A comparison of the glomerular field potentials before and after knife cuts that isolated the glomerular layer from the deeper layers of the olfactory bulb indicated that both N1 and N2 were produced by currents generated, for the most part, within the glomeruli. A laminar analysis of the field potential profiles evoked by olfactory nerve stimulation in standard medium, or in the presence of CNQX, showed that N1 and N2 reversed polarity in the external plexiform and mitral cell layers, suggesting that both components reflected synaptic responses in the distal, apical dendrites of mitral/tufted cells. Simultaneous field potential recordings in the glomerular layer and intracellular recordings in the mitral cell layer showed that: (i) N1 is associated with a brief, short-latency spiking activity of mitral cells, and (ii) N2 is associated with prolonged mitral cell spiking, since N2 and the late cell firing had similar time-courses, and both were blocked by bath applied DL-2-amino-5-phosphonovalerate. Application of the GABA(A) receptor antagonist bicuculline methiodide (10 microM) to standard medium selectively enhanced N2. The enhanced N2 was significantly reduced by DL-2-amino-5-phosphonovalerate. Strychnine, an antagonist of glycine receptors, had similar effects to those of bicuculline, but only at high concentrations that have been previously shown to block GABA(A) receptors; at low concentrations strychnine had no effect. The effects of all drugs tested were reversible. In the rat olfactory bulb, activation of the olfactory nerve evokes a kainate/AMPA receptor-mediated response in the distal, apical dendrites of mitral/tufted cells, followed by a slow N-methyl-D-aspartate receptor-mediated response which triggers prolonged discharge of mitral cells. GABA(A) receptor-mediated inhibition appears to suppress, preferentially, this N-methyl-D-aspartate receptor-mediated component. The presence of prolonged N-methyl-D-aspartate receptor-mediated postsynaptic activity at the primary synapses of the olfactory system may play a key role in olfactory processing by facilitating synaptic integration and plasticity.

Animals↗
PubMed1997

Discrete subregions of the rat midbrain periaqueductal gray project to nucleus ambiguus and the periambigual region.

We investigated the organization of projections from the rat midbrain periaqueductal gray to nucleus ambiguus and the periambigual region using retrograde and anterograde tract tracing techniques. Retrograde tracing results revealed that neurons that project to nucleus ambiguus arise from three discrete, longitudinally organized columns of neurons located in the supraoculomotor central gray, lateral and ventrolateral periaqueductal gray. Anterograde tracing studies demonstrated that projections from these three columns of periaqueductal gray neurons terminate with topographic specificity in nucleus ambiguus and the periambigual region. Double-labelling studies demonstrated that periaqueductal gray neurons terminate in close contiguity to cholinergic neurons in the compact, semicompact, loose and external formations of nucleus ambiguus. The present results suggest that projections from periaqueductal gray to nucleus ambiguus may mediate, in part, certain cardiovascular adjustments and vocalizations produced by stimulation of periaqueductal gray.

Animals↗
PubMed1997

Bronchoalveolar lavage findings suggest two different forms of childhood asthma.

BACKGROUND: It seems plausible that children with atopy and persistent asthma symptoms will, like their adult counterparts, have chronic airways inflammation. However, many young children with no other atopic features have episodic wheezing that is triggered solely by viral respiratory infections. Little is known as to whether airways inflammation occurs in these two asthma patterns during relatively asymptomatic periods. METHODS: Using a non-bronchoscopic bronchoalveolar lavage (BAL) procedure on children presenting for an elective surgical procedure, this study has investigated the cellular constituents of BAL fluid in children with a history of atopic asthma (AA) non-asthmatic atopic children (NAA) or viral associated wheeze (VAW). RESULTS: A total of 95 children was studied: 52 with atopic asthma (8.0 years, range 1.1-15.3, 36 male), 23 with non-asthmatic atopy (median age 8.3 years, range 1.7-13.6, 11 male) and 20 with VAW (3.1 years, range 1.0-8.2, 13 male). No complications were observed during the lavage procedure and no adverse events were noted post-operatively. Total lavage fluid recovered was similar in all groups and the total cell numbers were higher in the VAW group. Eosinophil (P < or = 0.005) and mast cell (P < or = 0.05) numbers were significantly elevated in the group with atopic asthma. CONCLUSIONS: During relatively asymptomatic periods there is on-going airways inflammation, as demonstrated by eosinophil and mast cell recruitment, in children with asthma and atopy but not in children with viral associated wheeze or atopy alone. This strongly suggests that there are different underlying pathophysiological mechanisms in these two groups of children who wheeze.

Adolescent↗
PubMed1997

A comparison of three standard methods of identifying mast cells in endobronchial biopsies in normal and asthmatic subjects.

Reported mast-cell counts in endobronchial biopsies from asthmatic subjects are conflicting, with different methodologies often being used. This study compared three standard methods of counting mast cells in endobronchial biopsies from asthmatic and normal subjects. Endobronchial biopsies were obtained from atopic asthmatic subjects (n = 17), atopic nonasthmatic subjects (n = 6), and nonatopic nonasthmatic control subjects (n = 5). After overnight fixation in Carnoy's fixative, mast cells were stained by the short and long toluidine blue methods and antitryptase immunohistochemistry and were counted by light microscopy. Method comparison was made according to Bland & Altman. The limits of agreement were unacceptable for each of the comparisons, suggesting that the methods are not interchangeable. Coefficients of repeatability were excellent, and not different for the individual techniques. These results suggest that some of the reported differences in mast-cell numbers in endobronchial biopsies in asthma may be due to the staining method used, making direct comparisons between studies invalid. Agreement on a standard method is required for counting mast cells in bronchial biopsies, and we recommend the immunohistochemical method, since fixation is less critical and the resultant tissue sections facilitate clear, accurate, and rapid counts.

Adolescent↗
PubMed1996

Activation of locus coeruleus enhances the responses of olfactory bulb mitral cells to weak olfactory nerve input.

The main olfactory bulb (MOB) receives a dense projection from the pontine nucleus locus coeruleus (LC), the largest collection of norepinephrine (NE)-containing cells in the brain. LC is the sole source of NE innervation of MOB. Previous studies of the actions of exogenously applied NE on mitral cells, the principal output neurons of MOB, are contradictory. The effect of synaptically released NE on mitral cell activity is not known, nor is the influence of NE on responses of mitral cells to olfactory nerve inputs. The goal of the present study was to assess the influence of LC activation on spontaneous and olfactory nerve-evoked activity of mitral cells. In methoxyflurane-anesthetized rats, intracoerulear microinfusions of acetyicholine (ACh) (200 mM; 90-120 nl) evoked a four- to fivefold increase in LC neuronal discharge, and a transient EEG desynchronization and decrease in mitral cell discharge. LC activation increased excitatory responses of mitral cells evoked by weak (i.e., perithreshold) nasal epithelium shocks (1.0 Hz) in 17/18 cells (mean Increase = 67%). The discharge rate of mitral cells at the time that epithelium-evoked responses were increased did not differ significantly from pre-LC activation baseline values. Thus, changes in mitral baseline activity do not account for the increased response to epithelium stimulation. These findings suggest that increased activity in LC-NE projections to MOB may enhance detection of relatively weak odors.

Acetylcholine↗
PubMed1996

Olfactory nerve stimulation activates rat mitral cells via NMDA and non-NMDA receptors in vitro.

The neurotransmitter(s) and receptors mediating excitatory transmission at the mammalian olfactory nerve-mitral cell synapse were investigated using extracellular recordings in rat olfactory bulb slices. Single shocks applied to the olfactory nerve elicited both a short latency and a delayed excitatory response in mitral cells. Both responses were blocked after bath application of kynurenic acid, a broad-spectrum glutamate receptor antagonist, or DNQX, a preferential non-NMDA receptor antagonist. The specific NMDA receptor antagonist AP5 selectively attenuated the delayed, but not the initial excitation. These results suggest that glutamate is the major excitatory transmitter in the mammalian olfactory nerve, and excites mitral cells via NMDA and non-NMDA receptors.

2-Amino-5-phosphonovalerate↗
PubMed1996

Dendrites of locus coeruleus neurons extend preferentially into two pericoerulear zones.

The intrinsic cytoarchitecture and neurochemical organization of the nucleus locus coeruleus have been characterized extensively, but there is little information about the organization of locus coeruleus neuronal processes extending outside of the nucleus proper. Light and electron microscopic immunocytochemical techniques were used to investigate the distribution of dopamine-beta-hydroxylase- or tyrosine-hydroxylase-labeled extranuclear processes in the rat pericoerulear region. The vast majority of these processes extended preferentially into two zones: (1) the pontine tegmentum medial and rostral to locus coeruleus, here termed the rostromedial pericoerulear region; and (2) a narrow region adjacent to the IVth ventricle caudomedial to locus coeruleus, designated here as the caudal juxtaependymal pericoerulear region. Far fewer labeled processes extended into the lateral and ventral pericoerulear regions. Seventy-seven percent of the labeled profiles in the pericoerulear region were dendrites. All labeled profiles in the rostromedial pericoerulear region and 94% of the labeled profiles in the caudal juxtaependymal zone were dendrites. By contrast, in the rostroventral pericoerulear region, 25% of the labeled profiles were axons. Locus coeruleus extranuclear dendrites were never presynaptic to other structures but were often contacted by several unlabeled presynaptic terminals. These results indicate that the dendrites of locus coeruleus neurons extend preferentially into two pericoerulear zones. Extranuclear dendrites in all pericoerulear regions receive extensive, nonnoradrenergic synaptic contacts. Thus, pericoerulear dendrites, particularly in the rostromedial and caudal juxtaependymal zones, are important sites for the integration of inputs to locus coeruleus neurons.

Animals↗