New phenotypic diversity associated with the mitochondrial tRNA(SerUCN) gene mutation.

We performed detailed clinical, histopathological, biochemical, in vitro translation and molecular genetic analysis in patients from two unrelated families harbouring the tRNA(SerUCN) 7472C-insertion mutation. Proband 1 developed a progressive neurodegenerative phenotype characterised by myoclonus, epilepsy, cerebellar ataxia and progressive hearing loss. Proband 2 had a comparatively benign phenotype characterised by isolated myopathy with exercise intolerance. Both patients had the 7472C-insertion mutation in identical proportions and they exhibited a similar muscle biochemical and histopathological phenotype. However, proband 2 also had a previously unreported homoplasmic A to C transition at nucleotide position 7472 in the tRNA(SerUCN) gene. This change lengthens further the homopolymeric C run already expanded by the 7472C-insertion. These data extend the phenotypic range associated with the 7472C-insertion to include isolated skeletal myopathy, as well as a MERRF-like phenotype.

Structural assessment of the urethral sphincter in women with urinary retention.

PURPOSE: The pathophysiology of urinary retention in women is generally unknown but a subgroup of women with urinary retention have been diagnosed as having so-called primary disorder of sphincter relaxation on the basis of an abnormal urethral sphincter electromyogram. It was suggested this sphincter overactivity could lead to work hypertrophy of the urethral rhabdosphincter and in this study we looked for any evidence of such muscle fiber hypertrophy. MATERIALS AND METHODS: In 9 women 18 to 45 years old (mean age 31.6) with urinary retention and overactive urethral sphincter electromyogram, light and electron microscopy were used to examine core needle biopsies of the urethral rhabdosphincter taken under transvaginal ultrasound control. Of the 9 patients only 5 biopsies processed for light microscopy and 4 processed for electron microscopy contained striated urethral muscle fibers. The results of these biopsies were compared to the morphology of a control specimen from a postmenopausal woman without a history of urinary retention. RESULTS: On light microscopy the urethral rhabdosphincter fiber diameter did not differ among patients (mean average 7.6 mum), was less than that reported in the literature (15 to 20), but did not differ from that of the control (mean 9.9). In all patients electron microscopy showed excessive peripheral sarcoplasm with lipid and glycogen deposition, and sarcoplasmic accumulation of normal mitochondria. These ultrastructural abnormalities were not seen in the control. CONCLUSIONS: To our knowledge this is the first morphological description of the urethral rhabdosphincter in a subgroup of women with urinary retention. Mean rhabdosphincter fiber diameter was approximately the same in patients and controls. This study does not support the previous theory that urethral sphincter overactivity in a subgroup of women with urinary retention leads to work hyperplasia of urethral rhabdosphincter fibers. An alternative hypothesis is suggested.

The role of the human bladder lamina propria myofibroblast.

OBJECTIVES: To describe the ultrastructure and relationship to nerves of the myofibroblast in the human bladder lamina propria, and discuss its possible role in bladder function, including sensing stretch, as the response of the bladder to stretch has been thoroughly investigated by afferent nerve recordings, but specialized stretch sensing organs have yet to be identified. MATERIALS AND METHODS: Flexible cystoscopic bladder biopsies were obtained from patients with detrusor hyper-reflexia and from controls. Systematic electron micrographs were obtained throughout the lamina propria, and the presence and location of cells with ultrastructural characteristics of myofibroblasts noted, together with their relation to surrounding nerves. RESULTS: Within the lamina propria there was a layer of cells with the cytological characteristics of both fibroblasts and smooth muscle cells, that included bundles of fine cytoplasmic filaments, dense bodies, linear arrays of subsurface vacuoles, and the presence of an interrupted basal lamina. This combination of features is characteristic of the myofibroblast. These cells had close contacts with unmyelinated axonal varicosities containing a mixture of clear and large dense-cored vesicles, or clear vesicles alone. CONCLUSIONS: There is a layer of cells with the ultrastructural characteristics of myofibroblasts within the human bladder lamina propria. Their close contacts with nerves containing both small clear, and small clear with dense-cored, vesicles implies they have both an efferent and an afferent nerve supply, possibly functioning as a bladder stretch receptor. Furthermore, because of their similarities with the interstitial cells of Cajal in the gut, which are claimed to modulate small intestinal function, we discuss other possible roles for bladder lamina propria myofibroblasts.

Human T cell leukaemia virus type I associated neuromuscular disease causing respiratory failure.

Polymyositis and inclusion body myositis have rarely been described in association with human T cell leukaemia virus type I (HTLV-I) infection. Most of such patients have coexisting HTLV-I associated myelopathy (HAM). Two patients with HTLV-I infection, myopathy, and respiratory failure are described. The muscle biopsy specimen of the first patient bore the histological features of inclusion body myositis and there was no evidence of concurrent myelopathy. The second patient had HAM, and her muscle biopsy showed non-specific myopathic and neuropathic changes. Both patients developed respiratory muscle weakness over eight years after diagnosis of myopathy, leading to hypercapnic respiratory failure requiring mechanical ventilatory support. Respiratory failure as a complication of HTLV-I associated myopathy has not previously been described.

A Pacinian corpuscle in the human bladder lamina propria.

The fortuitous finding of a complex Pacinian corpuscle within the lamina propria of the human urinary bladder is described. It consisted of a complex of encapsulated nerve endings within the areolar connective tissue of the lamina propria immediately adjacent to the inner aspect of the detrusor muscle. It showed no structural evidence of directional sensitivity and was associated on its outer aspect with small unmyelinated axons containing small clear and dense-cored vesicles. This appears to be the first report of an encapsulated nerve ending within the lining of the adult human urinary bladder.

Immunopathology of cerebral malaria: morphological evidence of parasite sequestration in murine brain microvasculature.

A murine model that closely resembles human cerebral malaria is presented, in which characteristic features of parasite sequestration and inflammation in the brain are clearly demonstrable. "Young" (BALB/c x C57BL/6)F(1) mice infected with Plasmodium berghei (ANKA) developed typical neurological symptoms 7 to 8 days later and then died, although their parasitemias were below 20%. Older animals were less susceptible. Immunohistopathology and ultrastructure demonstrated that neurological symptoms were associated with sequestration of both parasitized erythrocytes and leukocytes and with clogging and rupture of vessels in both cerebral and cerebellar regions. Increases in tumor necrosis factor alpha and CD54 expression were also present. Similar phenomena were absent or substantially reduced in older infected but asymptomatic animals. These findings suggest that this murine model is suitable both for determining precise pathogenetic features of the cerebral form of the disease and for evaluating circumventive interventions.

Fifty year follow-up of a patient with central core disease shows slow but definite progression.

The follow-up of a patient with central core disease (CCD) over 50 years showed that although initially the condition was moderately non-progressive, progression of a significant degree did eventually occur. Histopathological and electron microscopic data were available from muscle biopsies carried out at the ages of 19 and 55 years, and show a marked predominance of type 1 fibres with central cores in most fibres at both ages. The four mutations within the RYR1 gene described in association with CCD and three of the more common malignant hyperthermia-associated mutations within RYR1 were not present.

Nemaline myopathy: an unusual cause of ophthalmoparesis.

Ophthalmoparesis and ptosis are extremely rare in nemaline myopathy. A 45-year-old man with a long history of bilateral ptosis and a 1-year history of diplopia is reported. Leg and arm weakness and wasting had been present since childhood, with a very slow deterioration over time. On examination, there was nonfatigueable bilateral ptosis that was more marked on the right. There was diplopia on left gaze. Extraocular movements showed limitation of elevation and adduction of the right eye. There was bilateral facial weakness, as well as proximal and distal wasting and weakness in the arms and legs. Electromyography (EMG) showed a combination of myopathic and neurogenic changes. Triceps muscle biopsy showed small multiple collections of rod-like structures in > 50% of fibers. This patient presented with a clinical picture that did not primarily suggest nemaline myopathy. This case illustrates the heterogeneity of this disorder and the need for muscle biopsy to make an accurate diagnosis in patients with ptosis and progressive external ophthalmoparesis.

Myelination and remyelination of aggregate rat brain cell cultures enriched with macrophages.

We reported previously that accumulation of myelin basic protein (MBP) in foetal brain aggregate cultures is enhanced by supplementation with peritoneal macrophages. The present study demonstrates that the rate of MBP accumulation in macrophage-enriched cultures continues to increase over time unaccompanied by a matching increase in the oligodendrocyte marker cyclic nucleotide phosphodiesterase, while that of control cultures reaches a plateau. These observations are supported by electron microscopic evidence of cumulative numbers of myelinated axons in the aggregates over time and by enhanced expression of myelin protein genes in macrophage-enriched relative to control cultures. Aggregates demyelinate following short-term exposure to cytokines and antimyelin oligodendrocyte glycoprotein antibody, and MBP synthesis resumes following removal of demyelinating agents. Supplementation of cultures with macrophages influences the degree of myelin breakdown and remyelination, drawing attention to the role that macrophage-derived growth factors may play in myelinogenesis and myelin repair in inflammatory demyelinating disease.

A rapidly progressive adolescent-onset oculopharyngeal somatic syndrome with rimmed vacuoles in two siblings.

We describe 2 Greek siblings who developed a rapidly progressive oculopharyngeal somatic syndrome, at the ages of 11 and 14 years, with muscle biopsies showing rimmed vacuoles and, in 1 case, cytoplasmic and intranuclear tubulofilamentous inclusions 25 nm in diameter. Although a similar pattern of muscle involvement with rimmed vacuoles is described in autosomal dominant oculopharyngeal muscular dystrophy, the age of onset, the rapid progression of the symptoms, and the nature of the tubulofilaments distinguish this as a separate entity.

Ultrastructural localization of sympathetic axons in experimental rat sciatic nerve neuromas.

The ultrastructural localization of sympathetic axons was investigated in normal rat sciatic nerves and experimental sciatic nerve neuromas. The best ultrastructural localization of noradrenaline in the dense-cored vesicles of sympathetic axons was accomplished following pretreatment of rats with nialamide and 5-hydroxy dopamine, followed by fixation according to the modified chromaffin technique of Tranzer and Richards (1976). After such preparation, sympathetic axons containing 5-hydroxy dopamine-labelled dense-cored vesicles could be identified in normal sciatic nerve. Large accumulations of labelled dense-cored vesicles were also found in acute neuromas, up to 1 week after nerve section. Much smaller numbers of dense-cored vesicles could be identified in chronic neuromas from 2 to 3 weeks following nerve section. Sympathetic axons could also be identified following electron probe X-ray microanalysis of the tissue sections, using chromium detection as the marker for the noradrenaline-containing dense-cored vesicles. Unusual configurations of Schwann cell subunits, which enclosed myelinated fibres and sympathetic axon sprouts within the same basal lamina, were identified in the acute neuromas, 3-7 days after nerve section. Such configurations may be of relevance to the pathophysiological interaction which develops between sympathetic efferent and sensory fibres in peripheral nerve neuromas.

The proton NMR spectrum in acute EAE: the significance of the change in the Cho:Cr ratio.

We have studied by nuclear magnetic resonance spectroscopy the evolution of the proton spectral changes in acute experimental allergic encephalomyelitis. We found an in vivo elevation in the ratio between the peaks assigned to "choline containing compounds" (Cho) and creatine plus phosphocreatine (Cr). This was associated with an increase in choline, betaine, and phosphorylcholine (PC) as well as a reduction in N-acetylaspartate (NAA), aspartate, N-acetylaspartatylglutamate and inositol in vitro. Histological examination revealed inflammation with no evidence of demyelination or neuronal loss. We conclude that the increase in the ratio of Cho:Cr was due to an increase in the concentrations of PC, betaine, and choline in association with inflammation, and not as others have suggested, with demyelination. The reported reduction in NAA may be due to dysfunction of neurones rather than their loss.

Mitochondrial myopathy associated with chronic zidovudine therapy in AIDS.

One hundred and eighteen consecutively identified AIDS patients, 88 of whom received zidovudine (1000-1200 mg/day), were followed for 1 year to investigate prospectively the relationship between zidovudine and myopathy. Clinical and biochemical evidence of proximal myopathy was seen in 7 of 41 patients (17%) who had been receiving zidovudine for more than 270 days, but in none of those on short-term therapy and in none of the controls. Serum creatine kinase levels rose a mean of 76 days (range 34-187) before the onset of clinical signs. Creatine kinase returned to normal within 4 weeks of cessation of zidovudine and strength returned within 8 weeks, though loss of muscle bulk persisted. Chronic malaise, anorexia and nausea accompanied the myopathy and remitted within 8 weeks of stopping zidovudine. Muscle histology in four patients with myopathy showed fibre size variation with atrophic, necrotic and degenerating fibres and an absence of inflammation. Ultrastructural studies showed glycogen-packed sarcoplasm, lipid droplets and grossly giant mitochondria. These abnormalities improved substantially after stopping zidovudine. Similar but less marked changes were seen in a zidovudine treated patient without myopathy, but were absent in one AIDS patient not taking the drug. Long-term zidovudine therapy is associated with a mitochondrial myopathy and the constitutional features suggest that it is part of a wider disorder affecting cellular function in other tissues.

Metabolically dependent blood-brain barrier breakdown in chronic relapsing experimental allergic encephalomyelitis.

We have studied chronic relapsing experimental allergic encephalomyelitis (CREAE), a model of immune-mediated demyelination, using gadolinium (Gd)-enhanced magnetic resonance imaging in vivo and the blood-brain barrier (BBB) markers, lanthanum nitrate and Gd nitrate, histologically. In regions of the spinal cord showing Gd enhancement, there was evidence for vesicular transport as a mechanism of BBB breakdown in CREAE, shown by an increased number of endothelial vesicles containing lanthanide (lanthanum or Gd, whichever had been perfused) and deposition of tracer in the perivascular space; tight interendothelial junctions remained intact. Prior perfusion with 2,4-dinitrophenol, a metabolic inhibitor, suppressed the appearance of endothelial vesicles containing lanthanide and tracer in the perivascular space. We conclude that an important contribution to BBB breakdown in CREAE is mediated by a metabolic change in the endothelial cells associated with increased vesicular transport.

A fluorescence study of changes in noradrenergic sympathetic fibres in experimental peripheral nerve neuromas.

Experimental neuromas were produced in rats by sciatic nerve section and avulsion of the distal stumps. At intervals varying from 3 days to 8 weeks after nerve section, the developing neuromas were resected and processed for noradrenaline (NA) fluorescence microscopy by the sucrose-phosphate-glyoxylic acid (SPG) method. From serial longitudinal sections through the neuromas and the nerve proximally, counts of noradrenergic sympathetic axons were made, together with qualitative observations of axon sprouting and NA content. By 3 days after nerve section there was a massive sprouting of sympathetic axons, with increased NA content, particularly towards the distal tip of the neuroma. Axon counts remained high 1 week following section then fell to below normal levels at 2 weeks, returning towards normal 8 weeks after nerve section. These results are discussed in relation to the known pathophysiological interaction between sympathetic efferent and sensory afferent fibres, which develops in neuromas following nerve section.

Duration and selectivity of blood-brain barrier breakdown in chronic relapsing experimental allergic encephalomyelitis studied by gadolinium-DTPA and protein markers.

Gadolinium-DTPA (Gd-DTPA) enhancement seen with magnetic resonance imaging in chronic relapsing experimental allergic encephalomyelitis (CREAE) corresponded with sites of blood-brain barrier breakdown judged by traditional markers in areas of inflammatory demyelination. Duration of Gd-DTPA leakage for individual lesions in CREAE varied from 5 days to more than 5 wks. By contrast, in acute EAE leakage was of shorter duration (always less than 5 days). Selective enhancement was observed in CREAE lesions using Gd-protein markers. Gd-albumin enhancement was not always seen in areas of leakage of the smaller molecular weight compound Gd-DTPA. The addition of immunoglobulin to the gadolinium complex led to enhancement of lesions not seen with Gd-albumin alone. From the similarities between the histology and the patterns of Gd-enhancement in CREAE and multiple sclerosis, it is probable that Gd-enhancement reflects active inflammation (with or without demyelination) in the human disease.

The characterization of experimental gliosis by quantitative nuclear magnetic resonance imaging.

Quantitative nuclear magnetic resonance imaging techniques were used to study the development of astrocytic gliosis following a cortical freezing injury in 4 cats, and the findings compared with the histological and ultrastructural appearances of the affected white matter. In the first week after lesioning, the acutely oedematous white matter was apparent as a region of signal abnormality with very prolonged T1' and T2' relaxation times. Following resolution of this oedema, the images remained normal for a period of approximately three months. After this latent period they became abnormal once again, and the abnormal signal was most evident in images emphasizing differences between tissues in T1' and proton density, but not T2'. The light and electron microscopic appearances of the corresponding tissue were of astrocytic gliosis in each animal. Measurement of the relaxation times, T1' and T2', of the abnormal regions revealed an increase in T1' without a corresponding change in T2'. The T2' magnetization decay characteristics were consistently monoexponential, in contrast to the clearly biexponential T2 decay which has previously been demonstrated in the presence of acute vasogenic oedema. These findings suggest that the pattern of change of the relaxation times and T2 magnetization decay might provide a means of distinguishing between lesions in multiple sclerosis which are predominantly gliotic, and those containing significant amounts of oedema.