Triamcinolone and prednisolone affect contractile properties and histopathology of rat diaphragm differently.

Diaphragm atrophy and weakness occur after administration of massive doses of corticosteroids for short periods. In the present study the effects of prolonged administration of moderate doses of fluorinated and nonfluorinated steroids were investigated on contractile properties and histopathology of rat diaphragm. 60 rats received saline, 1.0 mg/kg triamcinolone, or 1.25 or 5 mg/kg i.m. prednisolone daily for 4 wk. Respiratory and peripheral muscle mass increased similarly in control and both prednisolone groups, whereas triamcinolone caused severe muscle wasting. Maximal tetanic tension averaged 2.23 +/- 0.54 kg/cm2 (SD) in the control group. An increased number of diaphragmatic bundles in the 5-mg/kg prednisolone group generated maximal tetanic tensions < 2.0 kg/cm2 (P < 0.05). In addition, fatigability during the force-frequency protocol was most pronounced in this group (P < 0.05). In contrast, triamcinolone caused a prolonged half-relaxation time and a leftward shift of the force-frequency curve (P < 0.05). Histological examination of the diaphragm showed a normal pattern in the control and 1.25-mg/kg prednisolone group. Myogenic changes, however, were found in the 5-mg/kg prednisolone group and, more pronounced, in the triamcinolone group. Selective type IIb fiber atrophy was found in the latter group, but not in the prednisolone groups. In conclusion, triamcinolone induced type IIb fiber atrophy, resulting in reduced respiratory muscle strength and a leftward shift of the force-frequency curve. In contrast, 5 mg/kg prednisolone caused alterations in diaphragmatic contractile properties and histological changes without fiber atrophy.

The use of rhythmic auditory cues to influence gait in patients with Parkinson's disease, the differential effect for freezers and non-freezers, an explorative study.

PURPOSE: To study the effect of rhythmic auditory cues on gait in Parkinson's disease subjects with and without freezing and in controls. METHOD: A volunteer sample of 20 patients (10 freezers, 10 non-freezers) and 10 age-matched controls performed five randomized cued walking conditions in a gait-laboratory. Auditory cues were administered at baseline frequency, at an increased step frequency of 10 and 20% above baseline and at a decreased step frequency of 10 and 20% below baseline. Mean step frequency, walking speed, stride length and double support duration were collected. RESULTS: Rhythmical auditory cueing induced speed changes in all subjects. Stride length was not influenced by rhythmical auditory cues in controls, whereas patients showed a larger stride length in the -10% condition (p < 0.01). Freezers and non-freezers showed the same response to rhythmical auditory cues. Within group analysis for stride length showed different cueing effects. Stride length decreased at the +10% condition for freezers (p < 0.05), whereas it increased for non-freezers. CONCLUSIONS: This study points to fact that physiotherapists might need to carefully adjust the cueing frequency to the needs of patients with and without freezing. On the basis of the present results we recommend to lower the frequency setting for freezers, whereas for non-freezers an increase of up to +10% may have potential therapeutic use.

Non-participation in predictive testing for Huntington's disease: individual decision-making, personality and avoidant behaviour in the family.

Subjective risk perception, perceived impact of Huntington's disease (HD), perceived benefits and barriers of predictive testing and personality characteristics of persons withdrawing from the predictive test programme for HD and of siblings of test applicants were studied in a mailed survey. The belief that important decisions do not need to depend on a test result and the anticipated inability to cope with a bad result played an important role in the decision not to be tested. Nevertheless half of the group who ever considered testing, still planned to undergo a test in the future. A comparison of tested and untested persons revealed that the first group is more likely to overestimate the risk than the second group, but that both groups did not significantly differ from each other regarding anxiety, ego strength and coping strategies. An intrafamilial analysis of tested and untested siblings confirmed these findings. The problems during data collection and the reasons for the dropout are an illustration of the avoidant behaviour regarding HD and the predictive test in many individuals and families.

Limited expansion of the (CAG)n repeat of the Huntington gene: a premutation (?).

Huntington's disease (HD) is an autosomal dominant disorder with choreic movements, psychiatric manifestations and cognitive dysfunction. Recently the IT15 gene on chromosome 4p has been identified containing an unstable and expanded trinucleotide repeat in patients with HD. We report on the characteristics of this repeat in 248 individuals from 41 Belgian HD families. The length of the expanded repeat was defined precisely and reproducibly on an ALF sequencer and correlated well with the age of onset (r = -0.72). Paternal transmission of the expanded repeat resulted on average in a significantly longer repeat length (+2.79 repeats) than maternal transmission (-0.29 repeats). (CAG)n repeat of a premutation (?) size was observed in this population with subsequent expansion in the disease range. Presymptomatic or prenatal testing using only linked markers may be problematic in these cases.

alpha 2-Macroglobulin expression in neuritic-type plaques in patients with Alzheimer's disease.

Because it has been suggested that alpha 2M could be involved in the generation of amyloid peptide, attention was given to a possible association of alpha 2M expression and amyloid accumulation in the brain. Therefore, we investigated the presence of the proteinase inhibitor alpha 2-macroglobulin (alpha 2M) in the cerebra of 4 patients with Alzheimer's Disease (AD). One case of a patient with Down's syndrome, 2 cases of patients with Dementia of the Lewy Body type (DLB), 1 case of an aged, clinically nondemented person who displayed many amyloid plaques, and 3 normal aged control brains were also studied. The results obtained by immunocytochemistry with monoclonal antibodies directed against two different epitopes of human alpha 2M showed an association of alpha 2M, only with neuritic-type plaques in patients with AD. No alpha 2M immunoreactivity was found in either preamyloid-type plaques or burned out-type plaques in AD, DLB, or aged nondemented controls. The results do not support a direct role of this proteinase inhibitor in the formation of amyloid. Because alpha 2M is observed to be associated with reactive microglia in the outer border of the neuritic plaques, the data suggest that alpha 2M could be a marker for an inflammatory cellular process in these neuritic plaques.

Functional localization and cortical architecture in the nine-banded armadilli (Dasypus novemcinctus mexicanus).

A functional map of the armadillo neocortex was produced by cortical stimulation and recording evoked potentials following somatic, auditory and visual stimuli. The results obtained were then correlated with the cortical architecture as revealed by Nissl, Golgi and myelin-stained sections. Cortex rostral to the supraorbital sulcus has a wide layer IV and is mostly silent, except for a motor eye field and a part of the tongue sensory region in its caudal part. Two types of motor-sensory cortex are present caudal to the supraorbital sulcus. Postsupraorbital I is mostly motor and has prominent pyramidal layers. Layer V is particularly well developed and in rostral sections its superficial zone is broken up into clusters similar to the solid "barrels" seen in layer IV of other species. Postsupraorbital II has less prominent pyramidal layers and layers II and III are organized into clusters. This region corresponds to the sensory area for the limbs and trunk and the partially overlapping (surface recordings) sensory and motor areas for head, snout and tongue. Digits and limbs are rostral to the trunk representation in both the sensory and motor "homunculi." Even though surface recording was employed, potentials evoked by visual stimuli could only be recorded from a small caudal area with a very thin layer IV. Although striate and peristriate areas appear similar in Nissl stained preparations, they can be readily differentiated in Weil stained sections. The stellate character of neurons in layer IV of the visual cortex is particularly apparent in Golgi material. Auditory evoked surface potentials were recorded from a broad oval region in the caudal lateral cortex which has a wide layer IV and aggregates of neurons in layers II and III. A Weil stain demonstrates inner and outer bands of Baillarger in this same region. The presumptive insular cortex is electrically silent to sensory stimulation and presents as a narrow band just dorsal to the rhinal fissure with indefinite cell lamination and little myelin.

Observations on the development of brainstem-spinal systems in the North American oppossum.

The North American oppossum is born 12 to 13 days after conception and and is available for 90 days or more in an external pouch where it can be observed and experimentally manipulated. It is of particular interest that the hindlimbs of the newborn opossum are very immature and remain immobile for a week or more after birth. Degeneration techniques reveal that immature brainstem axons are present within the marginal zone of the lumbosacral cord before hindlimb movements begin (our state I) and material processed for formaldehyde induced fluorescence shows that some of them transport monoamines. Several lines of evidence suggest that part of the fluorescent axons arise within the nucleus locus coeruleus. At this early stage the electron microscope reveals that all brainstem-spinal axons are small (0.1--0.4 micrometer in diameter) and unmyelinated. By the time random hindlimb movements can be observed (stage II), brainstem axons, including those transporting monoamines, can be demonstrated to have grown into limited areas of the intermediate zone of the lumbosacral cord and to arise from most of the areas contributing to them in the adult animal (horseradish peroxidase technique). Such axons are still immature and it is not yet clear that they have formed synaptic terminals. Brainstem axons continue to grow into the intermediate zone of the lumbosacral cord for some time and come to occupy all of their adult territories before thoracic transection produces obvious change in hindlimb motility (beginning of stage III). It is still another 20 days or so before thoracic transection produces spinal shock comparable to that in the adult animal. The relatively mature use of the hindlimbs and the full expression of spinal shock correlate with changes in the technique and survival time needed to demonstrate degenerating brainstem axons in experimental material.

The inferior olivary nucleus of the opossum (Didelphis marsupialis virginiana), its organization and connections.

Although the inferior olivary nucleus of the opossum is small, sections stained either for Nissl substance, normal axons or cholinesterase activity reveal distinct medial, dorsal and principal nuclei. The medial nucleus contains three major subdivisions (labelled a, b, c after Bowman and Sladek, '73) and a group of neurons which is comparable to the cap of Kooy. In contrast to the cat and monkey, the major portion of the "medial" nucleus (subgroup a) lies lateral to the principal nucleus in rostral sections. The dorsal nucleus can also be subdivided, as can the principal nucleus which contains distinct dorsal and ventral lamellae. A small area is identified which based on position and connections may conform to the dorsal medial cell group. The experimental portion of the study provides evidence for an olivary projection from the motor-sensory cortex and a massive input from the midbrain (red nucleus, pretectum, midbrain tegmentum). In addition, the opossum inferior olive receives fibers from the deep cerebellar nuclei (cerebellar feedback loops), the spinal cord and the dorsal column nuclei. Of particular interest is the finding that fibers from the nucleus cuneatus and nucleus gracilis have distinctly different olivary targets and that those from the nucleus gracilis, but not the cuneate nucleus, overlap (in part, at least) with the direct spinal fibers. Other examples of overlapping fields of terminal degeneration are present and are discussed. In general our results reveal that although certain relationships between the nuclear divisions are different, the opossum olive conforms well to that of placental mammals and provides a basic mammalian model for future experimental electron microscopic and physiological studies.

Progressive pontobulbar palsy with deafness: clinical and pathological study of two cases.

Among the hereditary affections of the nervous system associated with deafness, a rare condition called "progressive pontobulbar palsy with deafness" has been described. In this slowly progressive condition, hearing loss and vestibular are-flexia are almost always the first symptoms, occurring in late childhood or early adulthood. Only 18 cases-some sporadic, several familial-have been published without a full report of pathological findings. The clinical and pathological data of two new cases-one familial, one sporadic-are described here. There are differences from other forms of bulbar paralysis, lower motor neuron diseases, and some spinocerebellar hereditary affections. In view of the homochrony and homotypy in familial cases and the pathological findings, progressive pontobulbar palsy with deafness appears to be an abiotrophic process wih autosomal recessive inheritance.

Correlations between triplet repeat expansion and clinical features in Huntington's disease.

OBJECTIVE: To investigate possible correlations between the length of the (CAG)n trinucleotide repeat in Hungtington's disease gene IT15 and clinical features (age at onset, symptoms at onset, and mode of progression) in Huntington's disease. DESIGN: In 59 patients with Huntington's disease, the expansion of the (CAG)n trinucleotide repeat was determined and clinical data were obtained retrospectively. SETTING: The Center for Human Genetics, affiliated with a university hospital. PATIENTS: All patients belonged to an initial group of 248 individuals tested in an indirect predictive testing procedure. RESULTS: A good correlation was found between the expansion of the (CAG)n trinucleotide repeat and the age at onset (r = -.71). No correlation was found between the repeat length of the normal allele and the age at onset. No correlations were found between repeat expansion and other clinical features, such as the nature of the symptoms at onset (neurologic, psychiatric/cognitive, or both) and the mode of progression. CONCLUSION: Factors that determine the nature of symptoms at onset and the mode of progression of Huntington's disease seem to be operating independently of the (CAG)n trinucleotide repeat in gene IT15.

Growth properties and in vitro life span of Alzheimer disease and Down syndrome fibroblasts. A blind study.

A blind study was set up to examine the in vitro growth characteristics of skin fibroblasts from 2 individuals with and 9 at risk for familial Alzheimer disease, 4 individuals with sporadic Alzheimer disease, 18 with Down syndrome as well as 5 younger and 6 older controls. Several variables (biopsy size, number of explants, medium, passage procedure) were standardized. Two growth characteristics were examined quantitatively: (i) the actual in vitro replicating life span was determined by counting the number of cells plated the previous week at 500,000 cells/flask (cumulative population doublings); and (ii) the growth potential was examined by a colony size distribution assay. A difference from the age-matched controls in the growth characteristics of skin fibroblasts was only observed for two patients with and one older individual at risk for familial Alzheimer disease. The growth properties of skin fibroblast cultures from patients with sporadic Alzheimer disease or Down syndrome were not at variance with their age-matched controls. The decrease in the growth potential observed in the familial Alzheimer disease fibroblasts is however modest and needs confirmation. It is clear that the growth properties of skin fibroblasts, as examined in this study, do not provide a good marker for any form of Alzheimer disease, nor do they provide an appropriate in vitro system to study factors which may contribute to the etiopathogenesis of Alzheimer disease or Down syndrome.

Neuropathology of Huntington's chorea. Studies of the ventrobasal complex of the thalamus.

In seven cases of Huntington's chorea, the ventrolateral thalamus was studied by quantitative cytometry. A selective 50 percent atrophy of microneurons (internuncial cells) was found while the macroneurons did not show significant atrophy. Thalamic microneurons might be presynaptic and postsynaptic inhibitory cells. Their specific atrophy in Huntington's chorea thus could be related to the known decrease of gamma aminobutyric acid (GABA) in Huntington's chorea.

Globoid cell leukodystrophy: a family with both late-infantile and adult type.

We present a patient with adult-onset globoid cell leukodystrophy (GBL) who had almost complete deficiency of galactosylceramide beta-galactosidase. A brother of the index patient deteriorated neurologically and died at the age of 4, probably from the late-infantile form of the disease. In this family, two clinical types of GBL are probably different expressions of an identical genotype.

Early-onset Alzheimer's disease in 2 large Belgian families.

Familial Alzheimer's disease (FAD) is a dominantly inherited condition that may present with an early onset, and myoclonus occurs frequently in the course of the disease. We report clinical and neuropathologic data on 2 large Belgian families with FAD in which we obtained 17 autopsies of the CNS. In family A, each of 11 autopsies had the typical neuropathologic features of Alzheimer's disease (AD), and there were a few cerebellar plaques in the molecular layer. In family B, in addition to the typical characteristics of AD in 6 autopsies, there were numerous amyloid plaques in the cortical cerebellar layers. In both families, we immunostained the amyloid deposits for the A4 protein, and they were negative for prion-associated protein immunoreactivity.

Diffuse fasciitis after bone marrow transplantation.

This report describes a patient in whom the clinical, laboratory, and histologic features of diffuse fasciitis with eosinophilia developed several months after allogeneic bone marrow transplantation for acute lymphoblastic leukemia. Numerous reports detail the association between diffuse fasciitis and hematologic diseases; a patient in whom diffuse fasciitis developed in the setting of chronic graft-versus-host disease following bone marrow transplantation for acute leukemia is discussed. Treatment of the chronic graft-versus-host disease improved the symptoms of the diffuse fasciitis.

Production of tumor necrosis factor in spinal cord following traumatic injury in rats.

Production of tumor necrosis factor (TNF) in the spinal cord following traumatic injury has been studied. In these experiments, the level of TNF was examined in the homogenate of the spinal cord, cerebrospinal fluid (CSF) and serum (n = 56). TNF could be detected in the injured spinal cord but not in the normal spinal cord. The TNF level increased in the spinal cord after the injury. At the lesion site, a maximal TNF concentration was observed 1 h after the injury, and the TNF concentration remained at this level until 8 h after the injury. Thereafter, it decreased gradually. However, TNF still could be detected 72 h after the injury. No TNF could be detected in the CSF and serum, collected from rats both with and without spinal cord injury (SCI). This study thus suggests that TNF is produced locally in the spinal cord following traumatic injury, and this TNF production is caused by the injury. The present results also demonstrate that TNF production is an acute and rapid reaction in the spinal cord following traumatic injury.

Leu-3+ lymphocytes account for increased CSF cellularity.

Inflammatory conditions of the central nervous system (CNS) are often marked by an increase in lymphocyte number in the cerebrospinal fluid (CSF). In order to determine if changes in CSF cell numbers can alter T-lymphocyte subset composition in CSF or in blood, cell surface markers were evaluated in 25 CSF and paired blood samples from a variety of neurologically affected patients. T-cell subset levels in peripheral blood did not reflect subset levels in paired CSF samples. However, CSF samples with elevated cell numbers (greater than 3 cells/mm3) had significantly increased levels of Leu-3+ T-cells (P less than 0.001), but not Leu-2+ T-cells relative to CSF samples with low cell counts. These data suggest a selective increase in the Leu-3+ T-lymphocyte subset in CSFs with increased cellularity in patients with acute neurologic signs.

The use of lead as an ionic tracer for investigating routes of passive fluid transfer.

Several inorganic ions, including lead, barium, silver, and thallium, have been tested as possible tracers for demonstrating fluid-accessible channels in functional epithelia at the ultrastructural level. The most useful of the ionic tracers examined was the lead (plumbous) ion, administered for short time intervals (less than 2 min) and "captured" with phosphate used as the buffer in the fixative. Passive fluid and ion-accessible channels of rat parotid salivary gland have been examined with this method. At short tracer infusion times (0.5-1.0 min), localization of the tracer was primarily extracellular, although intracellular deposits were observed in the following sites: smooth membrane-delimited endocytic vesicles of both epithelial and connective tissue cells, inner Golgi cisternae, and occasional cisternae of rough endoplastic reticulum. The lead tracer readily penetrated tight junctions between parotid acinar cells but rarely passed through the tight junctions between intercalated duct cells and did not penetrate junctions of striated duct cells. Fat cells observed in the stroma of this gland were the only cells that exhibited lead tracer in the cytosol, suggesting that the plasmalemma of this cell type is more permeable to exogenous ions than the plasmalemma of other cell types present in this gland.

Immunocytochemical localization of tissue kallikrein in brain ventricular epithelium and hypothalamic cell bodies.

A specific monoclonal antibody against rat tissue kallikrein was used as the primary antibody for indirect immunoperoxidase staining of rat hypothalamus. Kallikrein was localized in the epithelial cells (ependyma) lining the third ventricle as well as in cell bodies of arcuate, supraoptic, paraventricular, and ventromedial nuclei.

Corticosteroid treatment and nutritional deprivation cause a different pattern of atrophy in rat diaphragm.

Triamcinolone (TR) causes type IIb fiber atrophy in the rat diaphragm, which is associated with changes in contractile properties. We investigated whether this is a direct effect of TR or the result of an accompanying loss of body and diaphragm weights. For 6 wk, adult rats received saline intramuscularly, TR (0.5 mg/kg im), or nutritional depletion (ND) that resulted in a similar (approximately 40%) reduction in body weight as TR. In these animals, the half-relaxation time of the diaphragm bundles increased, the force-frequency relationship shifted leftward, and the resistance to fatigue was increased. No histological changes were found in the ND diaphragm, in contrast to severe myogenic alterations in the TR diaphragm. Type IIb fiber cross-sectional area (CSA) in the TR diaphragm was reduced by 51%, whereas type I and IIa CSAs were unaffected. In the ND animals, the CSAs of type I, IIa, and IIb fibers were reduced by 31, 33, and 52%, respectively. Similar changes occurred in the deep part of the m. gastrocnemius. In conclusion, myogenic changes and selective type IIb fiber atrophy were caused by TR, whereas ND induced generalized fiber type atrophy without histological changes.