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Biomedical subjects

B J Smith

Publications and source records attributed to B J Smith.

At least 73 records · Page 4Linked to original sources

Use of 16S rDNA sequences as signature characters to identify Xylella fastidiosa.

The nucleotide sequences of 16S rDNAs (coding for the small subunit ribosomal RNAs) were used to identify Xylella fastidiosa, a nutritionally fastidious plant pathogenic bacterium. The near-complete 16S rDNAs from nine strains of Xyl. fastidiosa, including seven pathotypes and one strain of Xanthomonas campestris pv. campestris, were amplified through PCR with two conserved primers (forward primer 5'-AGA GTT TGA TCC TGG CTC AG-3' and reverse primer 5'-AAG GAG GTG ATC CAG CC-3') and sequenced. The 16S sequences were compared with all eukaryote and prokaryote DNA entries in GenBank database. A Xyl. fastidiosa 16S rDNA sequence, M26601, was determined to be the most similar to all the near-complete (1537 bp) and partial 5' end sequences from Xyl. fastidiosa, but not those from the Xanthomonas strain. A 20-bp oligonucleotide (5'-TTG GTA GTA ATA CCA TGG GT-3') was found to be highly characteristic of Xyl. fastidiosa. Since the 16S rDNA of Xyl. fastidiosa strains are highly homologous and characteristically different from other bacteria, including the most closely related Xanthomonas, 16S rDNA sequences can be used as signature characters to identify this bacterium.

Base Sequence↗

Heavy metal concentrations in surface sediments in a nearshore environment, Jurujuba Sound, Southeast Brazil.

Sixty-four surface sediment samples and seven cored samples were collected from the partially closed bay of Jurujuba Sound, an inlet of Guanabara Bay in Southeast Brazil. Analysis of metals, including Pb, Zn, Ni, Cu and Cr, shows levels consistent with those typically found in urbanised and industrialised estuarine environments. Metal enrichment is concentrated around the inshore margin of the Sound and is significantly in excess of background, geological concentrations observed in basal muds from the seven cores. In the absence of industrialisation within the steep, but limited catchment that feeds into the Sound, the metal enrichment, particularly of Pb, Zn and Cu, is ascribed to the uncontrolled discharge of untreated sewage waste and urban surface runoff. This has increased markedly since the beginning of rapid urbanisation following the linking of the area by bridge to Rio de Janeiro in 1974.

Journal Article↗

Pulmonary alveolar proteinosis: two contrasting cases.

Pulmonary alveolar proteinosis is a rare condition characterized by the abnormal accumulation of surfactant-like material within the alveolar spaces and distal bronchioles. Two cases with contrasting modes of presentation, course, and response to therapeutic whole lung lavage are described. Both cases were in hypoxaemic respiratory failure at the time the definitive diagnosis was made, and in both cases the diagnosis was made by segmental bronchoalveolar lavage following negative open lung biopsy. In neither was an underlying causative organism or agent identified. In one case the alveolar proteinosis developed in late pregnancy, a presentation that is previously unreported. Clinical improvement in this case required repeated whole lung lavages and was accompanied by a trend towards normalization of the ratios of surfactant protein-A and surfactant protein-B to disaturated phospholipid, ratios which may be useful as prognostic indicators. The response to therapeutic lavage was markedly different in the two cases, and it is postulated that this may relate to the fact that alveolar proteinosis is a heterogeneous disease and that the course and response to treatment may relate in part to the specific composition of the abnormal proteinaceous fluid.

Adult↗

Improved efficiency of site-specific copper(II) ion-catalysed protein cleavage effected by mutagenesis of cleavage site.

The peptide sequence (N)DKTH(C) was previously investigated as a site for efficient, specific cleavage of a fusion protein by cupric ions using a humanized gamma1 Fab' as a model protein. Here we show that conservative mutations to three of the residues in the introduced cleavage site resulted in cleavage sites that were significantly improved. They were cleaved more efficiently by Cu(2+), such that cleavage reactions could be shorter, of lower pH or at a lower temperature. Some were even found to be measurably cleaved by Ni(2+). Use of these new cleavage sequences along with cupric ions may provide a more rapid and less harsh method for cost-effective, large-scale proteolytic cleavage of fusion proteins and peptides.

Amino Acid Sequence↗

In vitro kinetics of coumarin 3,4-epoxidation: application to species differences in toxicity and carcinogenicity.

Coumarin, a natural product and fragrance ingredient, is a well recognized rat liver toxicant, and dietary administration at toxic dosages increased the incidence of rat cholangiocarcinomas and parenchymal liver-cell tumors in a chronic bioassay. Hepatotoxicity in rats is site- and species-specific, and is thought to result from the formation of coumarin 3,4-epoxide and its rearrangement product, o-hydroxyphenylacetaldehyde (o-HPA). The goals of the current study were to describe the in vitro kinetics of the metabolic activation of coumarin, and determine whether species differences in susceptibility to liver injury correlate with coumarin bioactivation determined in vitro. Coumarin 3,4-epoxidation was quantified via the formation of o-HPA in pooled hepatic microsomes from female B6C3F1 mice, male F344 rats, and individual humans (n = 12 subjects), and the apparent kinetic constants for o-HPA production were calculated using nonlinear regression and fitting to either a one-enzyme or two-enzyme model. Eadie-Hofstee analyses indicated that o-HPA formation was biphasic in both rat and mouse liver. Although the apparent high affinity K:(m) in rat and mouse liver microsomes was 38.9 and 47.2 microM, respectively, the overall rate of o-HPA formation was far greater in mouse than in rat liver microsomes. Furthermore, the total clearance (CL(int)) of coumarin via o-HPA formation in mouse liver microsomes was 4-fold greater than in rat liver microsomes. Since mice are relatively resistant to hepatotoxicity, the data indicated that rates of o-HPA formation in rat and mouse liver microsomes were not directly predictive of liver toxicity in vivo, and further suggested that o-HPA detoxification played a role in modulating coumarin-mediated toxicity. The current studies also indicated that coumarin 3,4-epoxidation in human hepatic microsomes was minimal. In human liver microsomes (n = 12), the kinetics of o-HPA formation were best described by a single enzyme model, with the K(m) for o-HPA formation ranging from 1320-7420 microM. In the most active human sample, the intrinsic clearance of coumarin via the 3,4-epoxidation pathway was 1/9 and 1/38 that of the rat and mouse, respectively. The in vitro kinetics of o-HPA formation, and in particular, the large quantities of coumarin required for o-HPA production in human liver microsomes, strongly suggest that humans are unlikely to produce toxicologically relevant concentrations of this metabolite following low level coumarin exposures.

Acetaldehyde↗

A direct brain interface based on event-related potentials.

Cross-correlation between a trigger-averaged event-related potential (ERP) template and continuous electrocorticogram was used to detect movement-related ERP's. The accuracy of ERP detection for the five best subjects (of 17 studied), had hit percentages >90% and false positive percentages <10%. These cases were considered appropriate for operation of a direct brain interface.

Adult↗

Ictal spiking patterns recorded from temporal depth electrodes predict good outcome after anterior temporal lobectomy.

PURPOSE: Investigators have shown that the presence of ictal spiking (IS) recorded from temporal depth electrodes is associated with mesial temporal sclerosis (MTS). We investigated the relation of IS to seizure control and pathology after anterior temporal lobectomy (ATL). METHODS: All patients undergoing intracranial ictal monitoring from a single institution since 1989 were identified. Those who did not undergo ATL or had postoperative follow-up of <1 year were excluded. All received at a minimum bilateral temporal depth electrodes. Ictal recordings were reviewed for the presence of IS, and the proportion of seizures with IS was determined for each patient. Outcome was determined by using Engel's classification. Surgical specimens were reviewed for pathology. Statistics used were chi2, Fisher exact test, and Wilcoxon rank sum. RESULTS: Forty patients with 571 seizures were reviewed. In 292 seizures from 32 patients, IS was seen. Outcomes were 24 class I (22 with IS), five class II (four with IS), three class III (one with IS), seven class IV (four with IS), and one lost to follow-up (with IS). Pathologic review revealed 25 with MTS, 22 of whom had IS. The presence of IS was associated with class I outcomes (p = 0.04), but not MTS (p = 0.06). Patients with class I outcomes had a significantly greater proportion of seizures with IS (mean, 0.58 +/- 0.3) compared with other outcomes (mean, 0.30 +/- 0.3, p = 0.02). CONCLUSIONS: The presence of IS and higher proportion of seizures with IS correlated with good seizure outcome after ATL. This information may be used in preoperative counseling.

Electrodes, Implanted↗

A 5-year prospective study of Astra single tooth implants.

AIM: To evaluate the AstraTech Implant ST (Molndal, Sweden) for single tooth replacement clinically and radiographically after 5 years in function. SUBJECTS AND METHOD: Fifteen patients (age range 16 to 48) with missing maxillary anterior teeth (6 central incisors, 8 laterals, 1 bicuspid) had four 13 mm and eleven 15 mm single tooth implants provided. All patients were seen at 4- to 6-monthly intervals for oral hygiene maintenance. Periapical radiographs using Rinn holders and a long cone technique were taken at the crown insertion and after 1 year, 3 and 5 years. RESULTS: No implant losses were observed in 14 of the 15 patients available for evaluation. No abutment screw loosening or soft tissue problems were observed. At crown insertion the mean bone level was 0.46 +/- 0.55 to 0.48 +/- 0.56 mm apical to the top of the implant neck and there were no statistically significant changes in the radiographic bone level over the 5 years of the study (0.36 +/- 0.37 to 0.43 +/- 0.46 mm at year 5). One crown was recemented after 18 months in function and 1 crown was replaced because of a fracture to the porcelain incisal edge. CONCLUSION: The Astra Tech Implants ST were highly successful in single tooth replacement and bone levels during 5 years of function were stable.

Adolescent↗

Promoting physical activity in general practice: a controlled trial of written advice and information materials.

OBJECTIVE: To investigate the impact of a simple written prescription for physical activity given by a general practitioner and the effect of supplementing this with mailed information materials about physical activity. METHODS: A controlled trial was conducted in 27 general practices in New South Wales, Australia. Subjects were sequential routine care patients between 25 and 65 years old. Controls (n = 386) were recruited first, and intervention subjects two weeks later. Intervention subjects were randomised to receive a prescription only (n = 380) or a prescription plus a mailed booklet (n = 376). Self reported physical activity levels were measured by interview at baseline, 6-10 weeks, and seven to eight months. RESULTS: By intention to treat, the average changes in minutes of total physical activity did not differ significantly between the groups. Inactive people in the prescription plus supplementary booklet group were significantly more likely than controls to report an increase in their physical activity by at least 60 min/week after 6-10 weeks (odds ratio 1.58, 95% confidence interval 1.06 to 2.35). No significant short term improvements in self reported activity were shown in the prescription only group. In the supplemented group, the proportion reporting an increase in physical activity to 3,344 kJ/week at 6-10 weeks was not significant, and neither intervention group showed significant increases in any of the outcome measures at seven to eight months by intention to treat. Treatment received analysis showed greater improvements in intervention groups, especially the prescription plus booklet group, in which the odds of inactive people in this group reporting increased activity became significant at seven to eight months. CONCLUSIONS: A prescription for physical activity from a general practitioner, supplemented by additional written materials, can lead to modest short term improvements in self reported physical activity levels among inactive patients. A prescription alone was found not to be effective.

Adult↗

Factors associated with hospital admissions and repeat emergency department visits for adults with asthma.

BACKGROUND: A small proportion of patients with asthma account for a disproportionate number of acute health service events. To identify whether factors other than severity and low socioeconomic status were associated with this disproportionate use, a prospective study was undertaken to examine management and psychosocial factors associated with increased risk for admission to hospital with asthma and repeat visits to the emergency department over a 12 month period. METHODS: A total of 293 patients with moderate or severe asthma managed at least in part at two teaching hospitals completed surveys of clinical status, acute events, sociodemographic, and psychological variables. RESULTS: Twenty three percent had a single admission to hospital and 16% had two or more hospital admissions. Twenty six percent had one emergency department visit and 32% had two or more visits to the emergency department. In a multiple logistic regression model, adjusted for age, sex, education and income, odds ratios (95% CI) for baseline factors associated with hospital admissions over the next 12 months were: moderate severity compared with severe asthma 0.6 (0.2 to 0.9); no hospital admissions in the past 12 months 0.1 (0.01 to 0.2); not possessing a written asthma action plan 4.0 (1.5 to 10.7); less use of an avoidance coping style 0.4 (0.3 to 0.7); lower preferences for autonomy in asthma management decisions 1.4 (0.96 to 2.0). Adjusted odds ratios (95% CI) for repeat emergency department visits were: moderate asthma severity 0.3 (0.1 to 0.8); current regular use of oral corticosteroids 10.0 (3.1 to 32.4); a hospital admission in the past 12 months 2.9 (1.8 to 4.8); not possessing a written asthma action plan 2.2 (1.1 to 5.6); less dislike of asthma medications 0.7 (0.5 to 0.9). CONCLUSIONS: In addition to factors relating to severity, not possessing a written asthma action plan, avoidance coping, and attitudes to self-management were related to acute use of health services in this at risk group. Interventions need to address or take these factors into account to reduce asthma morbidity.

Adolescent↗

Gross morphology and topography of the adult intestinal tract of the tilapian fish, Oreochromis niloticus L.

The intestinal tract of the Nile tilapia, Oreochromis niloticus L., follows a complex course involving multiple loops and coils arranged in a previously undescribed form. From cranial to caudal, five principal regions were identified and designated as the hepatic loop (HL), proximal major coil (PMC), gastric loop (GL), distal major coil (DMC), and terminal segment (TS). The first four of these regions each possessed a reversal flexure and thus could be divided into proximal and distal limbs. Only the terminal segment was straight and undivided. The PMC and DMC were disposed in a spiral, cone-shaped mass (spiral intestine) - their proximal and distal limbs are thus designated as centripetal and centrifugal limbs. These spiral limbs were arranged with each successive limb nested internal to the previous one. Beginning from the stomach, the complete course of the gut including designations of the subdivisions of the major regions was as follows: proximal limb of the HL, distal limb of the HL, centripetal limb of the PMC, centrifugal limb of the PMC, proximal limb of the GL, distal limb of the GL, centripetal limb of the DMC, centrifugal limb of the DMC, and the TS. Though the topographical relations of the various gut loops permitted ready identification of each, external surface features were so similar among the segments that extirpated segments of gut could not be identified as to region of origin. The nesting of successive intestinal loops of the spiral intestine in this fish is novel among patterns previously described, and also among the more intricate of those that have been described.

Animals↗

Health effects of daily indoor nitrogen dioxide exposure in people with asthma.

Household gas appliances produce nitrogen dioxide (NO2), which may be associated with an increase in symptoms in asthmatics. The relationship between indoor NO2 exposure, and respiratory symptoms in people with asthma was evaluated. Self-reported asthmatics (n=125) wore lapel badges that measured NO2 daily over 6 weeks at home. Outdoor pollutants, spores and meteorological parameters were measured daily, in addition to smoking status and demographic factors. Seven asthma symptoms were recorded in diaries, for analysis by same day and also with 1 day lag exposures, using a generalized estimating equation. Significant interactions were demonstrated between NO2 at age < or =14 yrs, with respect to the symptoms of chest tightness on the same day (odds ratio (OR): 1.29, 95% confidence interval (CI): 1.16-1.43) and with a 1 day lag (OR: 1.29, 95% CI: 1.14-1.46), breathlessness on exertion with a 1 day lag (OR: 1.13, 95% CI: 1.00-1.28), daytime asthma attacks on the same day (OR: 1.13, 95% CI: 1.02-1.26) night asthma attacks on the same day (OR: 1.16, 95% CI:1.03-1.30) and with a 1 day lag (OR: 1.15, 95% CI; 1.03-1.29) after adjustment for potential confounders. A significant interaction between NO2 and age 35-49 yrs was demonstrated for coughs with a 1 day lag (OR: 1.15, 95% CI: 1.01-1.31). Daily personal exposures to NO2 are associated with asthmatic symptoms in children.

Adolescent↗

Validity of a modified version of the Marks Asthma Quality of Life Questionnaire.

To produce a scale useful for individual clinical decision making, the Marks Asthma Quality of Life Questionnaire (AQLQ-M) was modified to a 22-item scale using a 7-point Likert response scale, and the validity of the new instrument was assessed. Adult asthma subjects with moderate to severe disease, recruited from two hospitals in Adelaide, Australia, were surveyed at baseline (n = 293), and at 3-month follow-up (n = 234). Cronbach's alpha for the Total scale of the modified AQLQ-M (MAQLQ-M) was 0.97 and all subscale values exceeded 0.90. Test-retest reliability values for all scales were between 0.88 and 0.93. All correlations between disease reference measures were statistically significant to at least the p < 0.01 level. Stronger associations were seen with symptom and self-rating scales than for lung function, medication usage, and health service utilization measures of outcome. The MAQLQ-M showed good discriminative ability for all asthma symptom categories and for different FEV1 values. Moderate, statistically significant associations were seen between changes in MAQLQ-M scores and clinical measures. Higher baseline MAQLQ-M scores were associated with lower risks over 12 months for hospital admissions (odds ratio, OR = 0.58) and repeated emergency department visits (OR = 0.47). The MAQLQ-M is a highly valid measure of asthma-related quality of life.

Adult↗

Asthma, inhaled corticosteroid use, and bone mass in prepubertal children.

The aim of this cross-sectional study was to describe the role of asthma, asthma severity, and medication usage in bone mineralization of prepubertal children. Asthma severity, medication usage, and physical activity were assessed by questionnaire and objective measures in 330 children. Bone densitometry and body composition were measured by dual-energy x-ray absorptiometry. Asthma ever was reported by 110 subjects (33%). A diagnosis of asthma was not associated with any deficit in bone mass, whereas usage of inhaled corticosteroids (ICS) in the last year (but not past use) was associated with deficits in bone in the total body (only after adjustment for confounders), particularly for doses of > or =400 microg/day. These observations support current recommendations with regard to ICS usage in children, but require confirmation in longitudinal studies.

Absorptiometry, Photon↗

APS, an adapter protein with a PH and SH2 domain, is a substrate for the insulin receptor kinase.

APS (adapter protein with a PH and SH2 domain) is the newest member of a family of tyrosine kinase adapter proteins including SH2-B and Lnk. We previously identified SH2-B as an insulin-receptor-binding protein and substrate [Kotani, Wilden and Pillay (1998) Biochem J. 335, 103-109]. Here we show that APS interacts with the insulin receptor kinase activation loop through its SH2 domain and insulin stimulates the tyrosine-phosphorylation of APS. Furthermore, the phosphorylation of activation-loop tyrosine residues 1158 and 1162 are required for this interaction.

Adaptor Proteins, Signal Transducing↗

Randomised controlled trial of general practice based asthma clinics.

OBJECTIVE: To compare the effects on asthma morbidity of asthma clinics based in general practice with standard general practice care. DESIGN AND SETTING: A randomised controlled trial in eight general practices. Patients, general practitioners and outcomes assessors were not blinded to treatment allocation. PARTICIPANTS: 195 patients with asthma aged 5-64 years; 191 completed the trial. INTERVENTION: Three asthma clinic sessions over six months involving nurse counselling, education about asthma management, spirometry and consultation with the general practitioner. MAIN OUTCOME MEASURES: Patients reporting days lost from work or school, number of days lost, the presence of morning or nocturnal asthma symptoms, use of an action plan, medication use, current smoking, hospitalisation, and emergency visits. RESULTS: Asthma clinics were associated with a greater reduction in nocturnal symptoms, an increase in the ownership of peak flow meters and an increase in the number of patients commencing or resuming smoking. Both control and intervention groups showed similar improvement in days lost from work or school, the presence of symptoms, use of an action plan and taking reliever medication. CONCLUSION: Our study does not show that asthma clinics are more effective than standard general practice care in reducing asthma morbidity. It is uncertain how much of the improvement in outcomes was due to the asthma clinic, the influence of the study itself upon patients and practitioners, or other factors, such as the tendency for a patient's asthma management to improve over time.

Absenteeism↗

Talin contains three similar vinculin-binding sites predicted to form an amphipathic helix.

Using recombinant talin polypeptides and an SDS/PAGE-blot overlay assay, we have previously identified three regions of talin that are involved in binding to vinculin [Gilmore, Wood, Ohanian, Jackson, Patel, Rees, Hynes and Critchley (1993) J. Cell Biol. 122, 337-347]. We have confirmed these observations by using a yeast two-hybrid assay and shown that talin residues 498-656, 852-950 and 1929-2029 are each capable of binding to vinculin residues 1-258. We have further defined the three vinculin-binding sites in talin to residues 607-636, 852-876 and 1944-1969; alignment of these sequences shows 59% similarity, although there are only two identical residues. Predictions of secondary structure indicate that this vinculin-binding motif forms an amphipathic alpha-helix. The hydrophobic face of helix 607-636 contains three aligned leucines (residues 608, 615 and 622), which show conservative substitutions in the other two sites. To test the possibility that this might constitute a leucine zipper involved in vinculin binding, we mutated each leucine residue to an alanine. The results showed that this leucine repeat is not essential to the interaction between talin and vinculin. We also used the yeast two-hybrid system to define further the talin-binding site within vinculin residues 1-258. C-terminal deletions made in accordance with exon boundaries showed that vinculin residues 1-167 are capable of interacting with each of the three vinculin-binding sites in talin. However, all N-terminal deletions abolished binding. The results suggest that the talin-binding site in vinculin has a relatively complex fold, whereas the vinculin-binding motif in talin is contained within a short linear peptide sequence that is repeated three times in the talin rod domain.

Amino Acid Sequence↗