Search PubMed⌕ Search

Biomedical subjects

R H Brown

Publications and source records attributed to R H Brown.

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

Heterogeneity of subcellular localization and electrophoretic mobility of survival motor neuron (SMN) protein in mammalian neural cells and tissues.

Spinal muscular atrophy is caused by defects in the survival motor neuron (SMN) gene. To better understand the patterns of expression of SMN in neuronal cells and tissues, we raised a polyclonal antibody (abSMN) against a synthetic oligopeptide from SMN exon 2. AbSMN immunostaining in neuroblastoma cells and mouse and human central nervous system (CNS) showed intense labeling of nuclear "gems," along with prominent nucleolar immunoreactivity in mouse and human CNS tissues. Strong cytoplasmic labeling was observed in the perikarya and proximal dendrites of human spinal motor neurons but not in their axons. Immunoblot analysis revealed a 34-kDa species in the insoluble protein fractions from human SY5Y neuroblastoma cells, embryonic mouse spinal cord cultures, and human CNS tissue. By contrast, a 38-kDa species was detected in the cytosolic fraction of SY5Y cells. We conclude that SMN protein is expressed prominently in both the cytoplasm and nucleus in multiple types of neurons in brain and spinal cord, a finding consistent with a role for SMN as a determinant of neuronal viability.

Animals↗
PubMed1998

Generation of a 3-Mb PAC contig spanning the Miyoshi myopathy/limb-girdle muscular dystrophy (MM/LGMD2B) locus on chromosome 2p13.

Miyoshi myopathy (MM) and limb-girdle muscular dystrophy subtype 2B (LGMD2B) map to the same region on chromosome 2p13. To facilitate the cloning of the defective gene causing these two diseases, we used a combination of chromosome walking and expressed sequence tag (EST) screening and identified 864 P1-derived artificial chromosomes (PACs) whose inserts map to the MM/LGMD2B candidate region and surrounding areas. Among them, 139 are from a chromosome 2-specific PAC library and 725 are from a total genomic PAC library. A 3-Mb contig spanning the candidate region for MM/LGMD2B was assembled. This contig contains 200 PACs, 10 known genetic markers, 5 new polymorphic markers, 57 sequence tagged sites (STSs) generated from PAC end fragments, and 4 random STSs. In addition, we mapped 24 ESTs to this contig and excluded 37 ESTs from the contig, thus eliminating them as candidate MM/LGMD2B genes. The high-resolution, sequence-ready PAC contig for the MM/LGMD2B region provides a backbone for the identification of the disease gene(s) and for clarification of the relationship between the two diseases.

Chromosomes, Human, Pair 2↗
PubMed1998

Mutations in the glutamate transporter EAAT2 gene do not cause abnormal EAAT2 transcripts in amyotrophic lateral sclerosis.

Recently, variant mRNA transcripts for the astroglial glutamate transporter EAAT2 have been detected in brain tissues of 60% of patients with sporadic amyotrophic lateral sclerosis (SALS). We have tested the hypothesis that the gene for EAAT2 may be defective in some ALS cases. In 16 familial ALS (FALS) pedigrees without mutations in SOD1, we failed to detect genetic linkage to the EAAT2 locus. We next characterized the genomic organization of the EAAT2 gene and used single-strand conformation polymorphism analysis of genomic DNA to identify one novel mutation in a single SALS patient and two novel mutations in 2 affected FALS siblings. In the SALS patient, the mutation substitutes serine for an asparagine that might be involved in N-linked glycosylation of the EAAT2 protein. In the 2 affected individuals in the FALS family, we detected both a mutation in the 5' end of intron 7 and a silent G --> A transition at codon 234 in exon 5. It remains unclear whether this intron 7 mutation is related to the defective mRNA splicing. These studies indicate that germline mutations in the EAAT2 gene are infrequent and do not explain the presence of variant mRNA transcripts of EAAT2 in more than one-half of ALS cases.

Amyotrophic Lateral Sclerosis↗
PubMed1998

Limited corticospinal tract involvement in amyotrophic lateral sclerosis subjects with the A4V mutation in the copper/zinc superoxide dismutase gene.

We examined 11 subjects with inherited amyotrophic lateral sclerosis (familial amyotrophic lateral sclerosis, FALS) associated with the most common copper/zinc superoxide dismutase 1 (SOD1) mutation, an alanine for valine substitution in codon 4 (A4V). Autopsies were performed on 5 subjects. The clinical and pathological findings are described and compared with those of 9 sporadic ALS (SALS) subjects. There was no clinical evidence of upper motor neuron (UMN) involvement in 10 FALS A4V subjects. All subjects had lower motor neuron (LMN) signs and electrophysiological evidence of denervation in at least three limbs. All SALS subjects had signs of both UMN and LMN involvement. Pathological studies found severe abnormalities of LMNs in all FALS and SALS subjects. UMN involvement was either absent or mild in the A4V SOD1 FALS subjects and severe in the SALS subjects. Pathological abnormalities in systems other than the motor neurons were more frequent in the FALS A4V subjects. This information suggests that current diagnostic criteria for ALS, requiring dinical evidence for both upper and lower motor neuron involvement, should be modified; ie, the diagnosis should be deemed established when there is evidence of denervation in three or more limbs and a mutation in the gene for SOD1, even without dinical signs of UMN involvement.

Adult↗
PubMed1998

Identification of two mutations and a polymorphism in the chloride channel CLCN-1 in patients with Becker's generalized myotonia.

Myotonia congenita is an inherited muscle disorder characterized by muscle stiffness and hypertrophy. Its clinical phenotype depends, in part, on whether it is inherited as a dominant or recessive trait, respectively designated Thomsen's disease or Becker's generalized myotonia (BGM). In either case, it is associated with abnormalities in the muscle currents that are linked to the gene (CLCN-1) on human chromosome 7q35 encoding the skeletal muscle chloride channel. Single-strand conformation polymorphism analysis was used to screen two families with the BGM for mutations in the CLCN-1 gene. Two new mutations were found (G 201ins and A317Q). The latter mutation has been previously described in Thomsen's disease.

Amino Acid Substitution↗
PubMed1998

Genetic fine mapping of the Miyoshi myopathy locus and exclusion of eight candidate genes.

Miyoshi myopathy (MM) is an early adult-onset, autosomal recessive disorder characterized by weakness and muscular atrophy starting in the distal muscles. The disease locus has been previously mapped by linkage analysis to chromosome 2p using the microsatellite marker D2S291. Initial haplotype analysis of markers in families from three different origins (North American, Japanese, and Tunisian) suggested that the MM gene is located in a 4-cM region flanked by markers D2S292 on the telomeric side and D2S286 on the centromeric side. To delineate critical recombination events revealing a more refined localization of the MM gene, we have determined the pattern of segregation of 12 marker loci in two consanguineous families of Tunisian origin. In this study we have: (1) detected recombination events with the disease locus in one family, placing the MM gene most likely between markers D2S443 (CHLC.GGAA4D07.1876) and D2S2109; (2) generated a yeast artificial chromosome contig that spans approximately 3.8 megabases and extends from marker D2S358 to marker D2S286; (3) physically mapped 21 polymorphic markers, 5 genes, 3 STSs, and 1 EST within this contig; (4) detected and mapped a new polymorphism within this interval, allowing us to further reduce the MM locus to a 360-kilobase segment; (5) mapped the gene for the cytoskeletal protein beta-adducin within the MM candidate region, failing to find a consistent pattern of mutation of this gene in our MM patients; (6) excluded seven other candidate myopathy genes from the Miyoshi locus.

Calmodulin-Binding Proteins↗
PubMed1998

Lack of association between apolipoprotein E genotype and sporadic amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a neuro-degenerative disorder with both sporadic and familial forms. Approximately 20% of autosomal dominant ALS is caused by mutations in the Cu/Zn superoxide dismutase (SOD1) gene. The causes of the remaining forms of ALS are unknown. The apolipoprotein E (APOE) gene is a known genetic risk factor for Alzheimer disease (AD), another neuro-degenerative disease. The APOE-4 allele increases risk and decreases age at onset in AD. Studies examining ALS and APOE have failed to show a significant effect of APOE on overall risk in ALS. Studies examining the effect of APOE-4 on site of onset in ALS (bulbar or limb) have been contradictory, with some studies showing an APOE association with bulbar onset and others showing no effect. Sample size was limited in these previous reports, particularly with respect to the number of bulbar onset cases (n = 33, 34 and 53). The present study examines a large collaborative data set of ALS patients (n = 363; 95 with bulbar onset) and age-matched neurologically normal controls. The results for these data showed no significant differences in the percentage of subjects with the APOE-4/4 and APOE-4/X genotypes (X = APOE-2 or APOE-3) when comparing cases and controls in both the overall data set or in the data set stratified by site of onset. Similarly, logistic regression analysis in the overall and stratified data set while controlling for sex showed no increase or decrease in risk of ALS associated with the APOE-4 allele. In addition, there were no significant differences in age at onset between patients with APOE-X/X, and APOE-4/4 or APOE-4/X genotypes, overall or stratified by site of onset. We conclude based on these data that the APOE gene is not a major genetic risk factor for site of onset in ALS.

Adult↗
PubMed1998

Refined mapping and characterization of the recessive familial amyotrophic lateral sclerosis locus (ALS2) on chromosome 2q33.

Amyotrophic lateral sclerosis (ALS) is a progressive degenerative neuromuscular disease that shows familial, autosomal dominant inheritance in 10%-15% of cases. Previous genetic analysis of one large family linked a recessive form of familial ALS (FALS-AR type 3) to the chromosome 2q33-35 region. Using additional polymorphic markers, we have narrowed the size of the linked region to approximately 1.7 cM by linkage and haplotype analysis. We have also established a yeast artificial chromosome contig across the locus that covers an approximate physical distance of 3 million bases. Based on this contig, genes and expressed sequences that map near the 2q33 region have been examined to determine whether they are located within this ALS2 candidate locus. Five identified genes and 34 expressed sequence tags map within the region defined by crossover analysis and merit further consideration as candidate genes for this disease.

Chromosome Mapping↗
PubMed1998

Dental fear with and without blood-injection fear: implications for dental health and clinical practice.

The relation between dental, blood and injection fear and oral health was examined in 936 New Zealand 18-year-olds. Of the approximately ten percent (n = 96) of the sample who reported a dental fear, 1 in 10 also reported a fear of blood and 53% a co-morbid fear of injections. Study members with dental fear alone or co-morbid dental and blood or injection fear had significantly worse oral health (i.e. greater caries experience) than a no-fear comparison group or individuals with blood-injection fear only. Further, individuals with dental and blood or injection fear had a significantly higher level of recent tooth decay than individuals with dental fear alone. Time since last dental treatment also tended to be highest in this group. Implications for dental health and practice were discussed.

Adolescent↗
PubMed1998

New techniques and developments in physiologic imaging of airways.

Recently, noninvasive imaging methods for assessing regional and individual airway responsiveness have been developed in animal models and applied to humans with obstructive lung disease. This article examines the technique of high-resolution CT scan and MR imaging and their applications to the imaging of airways.

Animals↗
PubMed1998

Dysferlin, a novel skeletal muscle gene, is mutated in Miyoshi myopathy and limb girdle muscular dystrophy.

Miyoshi myopathy (MM) is an adult onset, recessive inherited distal muscular dystrophy that we have mapped to human chromosome 2p13. We recently constructed a 3-Mb P1-derived artificial chromosome (PAC) contig spanning the MM candidate region. This clarified the order of genetic markers across the MM locus, provided five new polymorphic markers within it and narrowed the locus to approximately 2 Mb. Five skeletal muscle expressed sequence tags (ESTs) map in this region. We report that one of these is located in a novel, full-length 6.9-kb muscle cDNA, and we designate the corresponding protein 'dysferlin'. We describe nine mutations in the dysferlin gene in nine families; five are predicted to prevent dysferlin expression. Identical mutations in the dysferlin gene can produce more than one myopathy phenotype (MM, limb girdle dystrophy, distal myopathy with anterior tibial onset).

Adult↗
PubMed1998

Metabolic dysfunction in familial, but not sporadic, amyotrophic lateral sclerosis.

Autosomal dominant familial amyotrophic lateral sclerosis (FALS) is associated with mutations in the gene encoding Cu/Zn superoxide dismutase (SOD1). Previous studies have implicated the involvement of metabolic dysfunction in ALS pathogenesis. To further investigate the biochemical features of FALS and sporadic ALS (SALS), we examined SOD activity and mitochondrial oxidative phosphorylation enzyme activities in motor cortex (Brodmann area 4), parietal cortex (Brodmann area 40), and cerebellum from control subjects, FALS patients with and without known SOD mutations, SALS patients, and disease controls (Pick's disease, progressive supranuclear palsy, diffuse Lewy body disease). Cytosolic SOD activity, predominantly Cu/Zn SOD, was decreased approximately 50% in all regions in FALS patients with SOD mutations but was not significantly altered in other patient groups. Marked increases in complex I and II-III activities were seen in FALS patients with SOD mutations but not in SALS patients. We also measured electron transport chain enzyme activities in a transgenic mouse model of FALS. Complex I activity was significantly increased in the forebrain of 60-day-old G93A transgenic mice overexpressing human mutant SOD1, relative to levels in transgenic wild-type animals, supporting the hypothesis that the motor neuron disorder associated with SOD1 mutations involves a defect in mitochondrial energy metabolism.

Adult↗
PubMed1998

"U" curve association of blood pressure and mortality in hemodialysis patients. Medical Directors of Dialysis Clinic, Inc.

BACKGROUND: Hypertension may play an important role in the pathogenesis of the excess cardiovascular and cerebrovascular (CV) morbidity observed in hemodialysis patients (HD). However, the optimal blood pressure (BP) range for HD patients has not been defined. We postulated that there is a "U" curve relationship between BP and CV mortality. To explore this hypothesis we studied 5,433 HD patients in Dialysis Clinic Inc., a large not-for-profit chain, over a five year period. METHODS: Cox regression, with fixed and time-varying covariates, was used to assess the effect of systolic blood pressure (SBP) and diastolic blood pressure (DBP), pre- and post-dialysis, on CV mortality, while adjusting for age, gender, ethnicity, primary cause of end-stage renal disease, Kt/V, serum albumin, and antihypertensive medications. RESULTS: The overall impact of BP on CV mortality was modest. Pre-dialysis, neither systolic nor diastolic hypertension were associated with an increase in CV mortality. Post-dialysis, SBP > or = 180 mm Hg (RR = 1.96, P < 0.015) and DBP > or = 90 mm Hg (RR = 1.73, P < 0.05) were associated with increased CV mortality. Low SBP (SBP < 110 mm Hg) was associated with increased CV mortality, pre- and post-dialysis. CONCLUSIONS: The results suggest the presence of a "U" curve relationship between SBP post-dialysis and CV mortality in HD patients.

Adult↗
PubMed1998

Prevalence of latex allergy among anesthesiologists: identification of sensitized but asymptomatic individuals.

BACKGROUND: Occupational exposure to natural rubber latex has led to sensitization of health-care workers. However, the prevalence of latex allergy among occupationally exposed workers in American hospitals has not been reproducibly determined. The objectives of the current study were to determine the prevalence of and risk factors for latex sensitization among a cohort of highly exposed health-care workers. METHODS: Participants were 168 of 171 eligible anesthesiologists and nurse anesthetists working in the Department of Anesthesiology and Critical Care Medicine. A clinical questionnaire was administered, and testing was performed using a characterized nonammoniated latex reagent for puncture skin testing, a Food and Drug Administration-approved assay to quantify latex-specific immunoglobulin E antibody in serum, and, when required for clarification, a validated two-stage (contact-inhalation) latex glove provocation procedure. RESULTS: The prevalence of latex allergy with clinical symptoms and latex sensitization without clinical symptoms was 2.4% and 10.1%, respectively. The prevalence of irritant or contact dermatitis was 24%. The risk factors identified for latex sensitization were atopy (odds ratio, 14.1; 95% CI, 1.8-112.1; P = 0.012); history of allergy to selected fruits, such as bananas, avocados, or kiwis (odds ratio, 9.8; 95% CI, 1.6-61.9; P = 0.015); and history of skin symptoms with latex glove use (odds ratio, 4.6; 95% CI, 1.6-13.4; P = 0.006). CONCLUSIONS: The prevalence of latex sensitization among anesthesiologists is high (12.5%). Of these, 10.1% had occult (asymptomatic) latex allergy. Hospital employees may be sensitized to latex even in the absence of perceived latex allergy symptoms. These data support the need to transform the health-care environment into a latex-safe one that minimizes latex exposure to patients and hospital staff.

Adult↗
PubMed1998

Dental caries and changes in dental anxiety in late adolescence.

Little is known about changes in dental anxiety with ageing and their association with changes in oral health. This study examined the relationship between changes in dental caries experience and dental anxiety from 15 to 18 years of age among adolescent participants in the Dunedin Multidisciplinary Health and Development Study. Dental anxiety was estimated using the Corah Dental Anxiety Scale (DAS), and individuals with a DAS score of 13+ were identified as being dentally anxious. Dental examinations were performed on 649 individuals at ages 15 and 18, and a DMFS score was computed for each. Caries prevalence among those who were dentally anxious at both 15 and 18 years was significantly higher than for those who were not at either age. Regression analysis revealed that dental anxiety predicted caries incidence between ages 15 and 18 years. Dental anxiety is likely to be a significant predictor of dental caries experience, and may be a risk factor for dental caries incidence.

Adolescent↗
PubMed1998

The myth of maximal airway responsiveness in vivo.

A sine qua non of hyperresponsive airway disease in asthmatic subjects is the lack of a maximal response with increasing doses of aerosol agonist challenge. Normal subjects, however, often appear to exhibit an airway response plateau effect even when challenged with high concentrations of agonist. To investigate this question of maximal narrowing in individual airways in vivo, we used high-resolution computed tomography to visualize canine airways narrowed by two routes of agonist challenge. We compared airway narrowing induced by methacholine (MCh) via the conventional aerosol route to that caused by local atomization of MCh directly to individual airways. Our results showed that, with aerosol challenge, airway responses never reached a truly flat plateau even at the highest possible nebulizer concentrations. Airway closure was never observed. However, when MCh was delivered directly to the airway luminal surface, airways could be easily narrowed to complete closure at modest (10 mg/ml) agonist concentrations. Thus neither the elastic recoil of the lung nor limitations of smooth muscle shortening can be responsible for the apparent plateauing of dose-response curves. We suggest that the plateau results from limitations associated with the delivery of high concentration of agonists via the aerosol route.

Aerosols↗
PubMed1998

Individual canine airways responsiveness to aerosol histamine and methacholine in vivo.

Inhalational challenges to histamine and methacholine (Mch) are commonly used to measure airway reactivity. In this study, we investigated the serial distribution of individual airway responsiveness in vivo following stimulation with histamine or methacholine by directly measuring airway area with high resolution computed tomography (HRCT). Anesthetized dogs were challenged on separate days to increasing aerosol concentrations of histamine or Mch. Airway areas of 13 or 14 individual airways in each dog ranging in size from 1.1 to 16.9 mm in diameter at baseline were measured. Dose response curves for each agonist showed a similar responsiveness to each agonist over all airway sizes measured. There was a slightly greater individual airway responsiveness to Mch throughout the airway tree, and this greater responsiveness was also present in those airways that are responsible for the decreased Cdyn observed with Mch challenge. We conclude that in vivo aerosol challenges with histamine or methacholine do not lead to preferential construction of large or small airways.

Aerosols↗