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Biomedical subjects

I G Mayhew

Publications and source records attributed to I G Mayhew.

At least 19 recordsLinked to original sources

Interstitial cells of Cajal and electrical activity of smooth muscle in porcine ileum.

AIM: To identify the interstitial cells of Cajal in the porcine ileum for the first time immunohistochemically and to examine the electrical properties of intestinal smooth muscle in the same region. METHODS: In vitro intracellular microelectrode recordings were made from smooth muscle cells in cross-sectional preparations from abattoir-derived healthy porcine ileum. Immunohistochemical labelling of interstitial cells of Cajal was performed using an anti-Kit antibody. RESULTS: Slow waves were recorded in the circular muscle layer of all ileal preparations. The mean resting membrane potential of smooth muscle cells was -61.0 +/- 1.3 mV. Slow waves had a mean amplitude of 8.5 +/- 0.5 mV, a frequency of 9.9 +/- 0.1 cycles per minute and a duration of 5.6 +/- 0.1 s. A waxing and waning pattern of slow wave activity was occasionally observed. In addition, higher frequency spiking activity associated with contractions was observed in some recordings. The L-type calcium channel blocker nifedipine abolished both the spiking activity and the contractions, but had no significant effect on slow wave characteristics. Current-injection manipulation of the resting membrane potential had no effect on slow wave amplitude, frequency or duration. Kit-immunoreactive interstitial cells of Cajal were identified in the ileal samples and were present in the region of the myenteric plexus and in the circular and longitudinal muscle layers. CONCLUSION: This study recorded slow waves in vitro and demonstrated immunohistochemically the presence of interstitial cells of Cajal in the normal porcine ileum. This study forms a basis for future physiological and pathophysiological comparative studies of intestinal motility.

Animals↗

Horses on pasture may be affected by equine motor neuron disease.

REASONS FOR PERFORMING STUDY: Equine motor neuron disease (EMND) was diagnosed in 3 horses maintained on lush, grass-based pasture. This contrasted with North American studies which identified limited or no access to green herbage as an important risk factor for EMND. HYPOTHESIS: Grazing horses that have an apparently adequate intake of pasture herbage to meet normal equine vitamin E requirements can develop EMND. METHODS: Owners of 32 European horses diagnosed with EMND completed a questionnaire regarding intrinsic, managemental, nutritional and environmental factors that could potentially be risk factors for EMND, and also regarding clinical signs, treatments and case outcome. Plasma/serum vitamin E data for these horses were supplied by the veterinarians. No control population was studied. RESULTS: Thirteen of 32 horses (termed the 'grazing' group) had part- or full-time access to grass-based pasture at the onset of EMND (median duration at pasture 12 h/day, range 3-24 h). Five of these horses were at pasture for at least 235 h/day at the onset of EMND, 2 of which were at pasture for at least 23.5 h/day throughout the year. Despite grazing, all these horses had a low vitamin E status. The remaining 19 horses resembled those cases reported from North America, in that they had no or limited access to pasture. CONCLUSIONS AND POTENTIAL RELEVANCE: A diagnosis of EMND should not be discounted on the basis that a horse has access, even full-time, to lush grass-based pasture. Inadequate vitamin E intake was probably not the sole cause of either the EMND or the low vitamin E status in the grazing horses; the latter was probably the result of abnormal bioavailability or excessive utilisation of vitamin E.

Animal Feed↗

Investigation and management of a cluster of cases of equine retained fetal membranes in Highland ponies.

Four at-risk Highland ponies on the same premises all retained their fetal membranes in the same breeding season. The ponies were treated with a combination of oxytocin, infusion and distension of the allantochorionic sac with fluid, gentle traction in one case, and supportive therapy. The possible causes of the condition were investigated with clinicopathological tests and analyses of the herbage for its species composition and fungal contamination. No obvious possible causative abnormalities were identified apart from a low total serum calcium level in one case. No fescue grass was found and no pathogenic fungi were detected.

Animals↗

Observations on the interstitial cells of Cajal and neurons in a recovered case of equine dysautonomia (grass sickness).

This paper describes the histopathological and immunohistochemical changes in the central, autonomic and enteric nervous systems in a well-documented case of equine dysautonomia (ED), after the animal had recovered without significant residual clinical signs. Evidence of neuronal degeneration, such as neuronal chromatolysis, glial scars or a decrease in density of neurons, was not observed in the midbrain, pons, medulla oblongata or spinal cord, including the nuclei of cranial nerves III, V, VII, X and XII. In addition, no evidence of muscle denervation or re-innervation, such as group atrophy of clusters of cells or grouping of cells of similar size in the triceps muscle, was observed. These findings support the hypothesis that the chromatolytic motor neurons may not progress to cell death in ED. The autonomic ganglia showed evidence of previous neuronal loss. In the myenteric and submucosal plexuses of the ileum there was significant loss of enteric neurons. However, in the ileum, a continuous network of interstitial cells of Cajal (ICC) was observed by means of immunohistochemical techniques. It is possible that recovery without significant residual clinical signs was associated with lack of damage to, or more likely recovery of, the ICC, which have a pacemaker function in the intestinal tract.

Animals↗

Radiographic anatomy of the soft tissue attachments of the equine stifle.

REASONS FOR PERFORMING STUDY: Radiography is a very important aspect of equine stifle imaging. The precise radiographic anatomy of the soft tissue structures of the equine stifle has not been described previously. OBJECTIVE: To describe the anatomical relationship between sites of attachment of soft tissue structures of the equine stifle and their locations on standard radiographic views. METHODS: The sites of bony attachments of the tendons, ligaments and fibrous portion of the joint capsules of equine stifles were determined by gross dissection. These sites of attachment were transposed onto one set of bones deprived of soft tissue and mapped using radiopaque markers. This specimen was then radiographed in the standard radiographic projections (lateromedial, caudocranial and caudal 60 degrees lateral-craniomedial oblique) to determine the position of the attachment sites on the radiographs. RESULTS: Two radiographic maps were drawn per radiographic projection, one for the attachment sites of the ligaments and tendons and one for the attachment sites of the joint capsules. CONCLUSIONS AND POTENTIAL RELEVANCE: The radiographic maps of the precise position of the soft tissue attachments of the tendons, ligaments and joint capsules of the equine stifle should assist interpretation of equine stifle radiographs.

Animals↗

The development and distribution of the interstitial cells of Cajal in the intestine of the equine fetus and neonate.

This study set out to determine the pattern of development and distribution of the interstitial cells of Cajal (ICC) in the intestinal tract of the equine fetus and neonate. Intestinal tissue samples from 12 naturally aborted equine fetuses and three euthanized neonates were collected and fixed in formalin prior to applying standard immunohistochemical labelling techniques targeting the c-Kit protein of the ICC. At 6 months of gestation, a network of ICC was present in the myenteric plexus region of both the small and the large intestine. ICC were also present within the circular muscle layer. In the large intestine, a proximal to distal gradient of distribution was evident, with few ICC observed in the more distal parts of the large intestine in the younger fetuses compared with the near-term animals. A transmural gradient of distribution was also evident within the large intestine, with the most luminal part of the muscularis externa being the last area to be colonized by ICC. This region did not appear fully developed until the early neonatal period. An increased density of ICC was noted throughout the large intestine in the regions of the taenial bands in all animals. This study is the first to describe ICC development and distribution in the equine fetus and neonate.

Animals↗

Interstitial cells of Cajal (ICC) in equine colic: an immunohistochemical study of horses with obstructive disorders of the small and large intestines.

REASONS FOR PERFORMING STUDY: The gastrointestinal pacemaker cells, the interstitial cells of Cajal (ICC), have been implicated in several human gastrointestinal dysmotility syndromes. Recently, the involvement of these cells in equine gastrointestinal diseases has been investigated in cases of equine grass sickness where a significant reduction in ICC density was observed. OBJECTIVE: To investigate ICC density in equine obstructive gastrointestinal disorders using immunohistochemical labelling methods. METHODS: Intestinal samples were analysed from 44 horses undergoing exploratory surgery for colic and from 11 control animals subjected to euthanasia for conditions not related to the gastrointestinal tract. Immunohistochemical labelling of ICC was carried out using an anti-c-Kit antibody. Two independent observers assessed ICC density using a semiquantitative grading system. RESULTS: There was a significant reduction in ICC density in horses with large colon disorders compared to the controls (P<0.01). Horses with strangulating lesions of the small intestine showed no difference when compared to the controls. CONCLUSIONS: There was a reduction in ICC density in horses with large intestinal disorders. POTENTIAL RELEVANCE: The reduction in ICC density may be associated with the clinical findings as well as recurrent colic episodes observed in a number of these cases. This immunohistochemical study provides a basis for future functional electrophysiological investigations to determine the precise effect of ICC reduction on equine intestinal motility.

Animals↗

Clinical anatomy of the equine sphenopalatine sinus.

REASONS FOR PERFORMING STUDY: Disorders of the equine sphenopalatine sinus, including empyema and neoplasia, have been reported to cause damage to cranial nerves II and V. However, the clinical anatomy of these sinuses is not well described in horses. OBJECTIVE: To examine the anatomy of the sphenopalatine sinuses in a range of equidae and, in particular, to examine the relationship of these sinuses to adjacent major nerves and vessels. METHODS: The anatomy of the sphenoidal and palatine paranasal sinuses was examined in 16 equidae, primarily using transverse skull sections. Relevant structures were documented and photographed. RESULTS: There was much variation between individual horses in sphenopalatine sinus anatomy. The sphenoidal sinuses were small in young horses and appeared to become larger and more complex with age. Variation was present in the extent that the sphenopalatine sinus extended into the basisphenoid bone. The septum dividing left and right sphenoidal sinuses was frequently not midline, but was intact in all cases. The sphenoidal and palatine sinuses communicated in most horses. In such cases, what could accurately be termed the (combined) sphenopalatine sinuses usually drained directly into the caudal maxillary sinuses. Additionally, in 5 out of 16 cases, some compartments of the sphenoidal sinus also drained into the ethmoidal sinus. The dorsal and lateral walls of the sphenoidal sinus were very thin and directly adjacent to cranial nerves II, III, IV, V and VI and major blood vessels. CONCLUSIONS: The equine sphenoidal and palatine sinuses are very variable in their anatomy, but are always in close proximity to multiple cranial nerves and major blood vessels. POTENTIAL RELEVANCE: Many cranial nerves and blood vessels could be damaged with disorders involving the sphenopalatine sinus, potentially causing major and variable neurological syndromes, haemorrhage and extension of sepsis.

Age Factors↗

Clinical and pathological observations in English cocker spaniels with primary metabolic vitamin E deficiency and retinal pigment epithelial dystrophy.

Fifteen English cocker spaniels with confirmed vitamin E deficiency were examined physically, ophthalmologically and neurologically. Eleven of them had clinical signs of neurological dysfunction which included ataxia, proprioceptive deficits, abnormal spinal reflexes and muscle weakness. In the two dogs examined histopathologically there was central neuronal fibre degeneration with prominent neuroaxonal dystrophy, particularly within the sensory relay nuclei of the brainstem, and one of the dogs had severe intestinal lipofuscinosis.

Animals↗

Changes in nasal mucosal innervation in horses with grass sickness.

REASONS FOR PERFORMING STUDY: Equine grass sickness is a dysautonomia characterised by widespread destruction of autonomic ganglia, resulting in the clinical signs of dysphagia, constipation, profuse sweating, tachycardia, rhinitis sicca and high mortality rate. Rhinitis sicca is a common finding in horses with the chronic form and we have postulated that alterations in autonomic innervation of the nasal mucosa might underlie this clinical presentation. OBJECTIVES: In this study, the expression and distribution of nerve fibres immunoreactive for calcitonin gene-related peptide (CGRP), substance P (SP), the general neuronal marker protein gene-product 9.5 (PGP 9.5; ubiquitin) and the intermediate neurofilaments (PAN-N; neurorfilaments L, M and H) in the nasal mucosa of normal horses (n = 10) and horses with EGS (n = 18; acute n = 8, subacute n = 3, chronic n = 7) was assessed. METHODS: Innervation density and distribution was investigated in the different groups using standard immunohistochemical techniques. RESULTS: Significant differences were noted when comparing the density and distribution of nerve fibres immunoreactive for PGP 95 and PAN-N, with PGP 95 consistently giving better staining in all groups and at all sites in the nasal mucosa. An apparent increase in the density of innervation was noted for acute vs. normal cases. A significant reduction in the density of innervation was noted only with PAN-N when comparing normal horses and acute cases with the chronic group (P < 0.05). CGRP and SP immunoreactive nerve fibres were typically most abundant in the epithelial and subepithelial layers, but the quality of staining and nerve fibre density was greater for SP, achieving significant difference in several comparisons. The density of innervation for SP was significantly reduced in the chronic group compared to the normal and acute groups (P < 0.01). A significant decrease was noted for CGRP only for the acute and chronic groups (P<0.05). CONCLUSIONS: These results demonstrate a reduction in the expression of the sensory neuropeptides in nasal mucosal innervation as a consequence of equine dysautonomia, and may underlie the clinical presentation of rhinitis sicca noted with this disease. POTENTIAL RELEVANCE: Nasal biopsy may be of use in antemortem diagnosis of grass sickness and identification of mucosal denervation; and might also be useful in the treatment of rhinitis in EGS cases.

Acute Disease↗

Systemic concentrations of antioxidants and biomarkers of macromolecular oxidative damage in horses with grass sickness.

REASONS FOR PERFORMING STUDY: The aetiopathogenesis of equine grass sickness (EGS) is unknown. The role of free radical-mediated neuronal damage has not previously been investigated in this condition. OBJECTIVES: To investigate the potential contribution of oxidative damage and antioxidant status to neurodegeneration in EGS. METHODS: Systemic levels of surrogate biomarkers were determined in 10 horses with acute EGS and in 2 control populations; 10 healthy horses co-grazing with the 10 EGS horses at the onset of clinical disease, and 10 healthy mares grazing where EGS has not been reported. RESULTS: EGS horses had alterations in levels of several antioxidants, consistent with oxidative stress, the acute phase response and/or the secondary metabolic complications of EGS. EGS horses had elevated plasma dihydroxyphenylalanine (DOPA) levels. CONCLUSIONS: The elevated DOPA levels probably reflected a generalised disturbance of catecholamine metabolism rather than increased DOPA production via free radical-mediated oxidation of tyrosine. However, there was no evidence of systemic macromolecular oxidative damage. POTENTIAL CLINICAL RELEVANCE: Further work is required to determine whether macromolecular oxidative damage occurring at the neuronal level contributes to EGS.

Animals↗

In vitro microelectrode study of the electrical properties of smooth muscle in equine ileum.

Intracellular microelectrode recordings were made from smooth muscle cells in cross-sectional preparations of equine ileum, superfused in vitro. Membrane potential oscillations and spike potentials were recorded in all preparations, but recordings were made more readily from cells in the longitudinal muscle layer than from cells in the circular layer. The mean (se) resting membrane potential (RMP) of smooth muscle cells in the longitudinal muscle layer was -51.9 (1.2) mV, and the membrane potential oscillations in this layer had a mean amplitude of 4.8 (0.4) mV, a frequency of 9.0 (0.1) cycles per minute and a duration of 5.8 (0.2) seconds. The membrane potential oscillations were preserved in the presence of tetrodotoxin. A waxing and waning pattern of membrane potential oscillation activity was observed. Nifedipine abolished the spiking contractile activity of the smooth muscle, did not abolish the membrane potential oscillations but did alter their temporal characteristics.

Action Potentials↗

Central neuropathology of equine grass sickness.

Equine grass sickness (EGS) is an acquired disease of unknown aetiology affecting horses kept at grass. The disease is characterised by postganglionic sympathetic and parasympathetic neuronal pathology and is categorised as a dysautonomia. This study undertook a systematic examination of brain stem cranial nerve nuclei in 59 cases of EGS. Pathology consisting of neuronal chromatolysis was most consistently noted in the lower motor neurons of the general visceral efferent nucleus of CN III and X and the general somatic efferent nuclei of CN III, V, VII and XII. The prevalence of chromatolysis differed significantly between the diagnostic categories and was inversely related to the age of the animal. Duration of disease had a small but significant negative effect on the proportion of nuclei with chromatolytic neurons in chronic cases. The distribution of chromatolytic neurons is unlike that reported in any other equine or human disease; however, it appears to be equivalent to that in other animals with primary dysautonomias. EGS should be classified as a multisystem disease.

Age Factors↗