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Biomedical subjects

M A Osborne

Publications and source records attributed to M A Osborne.

At least 19 recordsLinked to original sources
PubMed2006

Multi-element survey of wild edible fungi and blackberries in the UK.

A survey of 12 metals including lead (Pb), copper (Cu), cadmium (Cd), mercury (Hg), arsenic (As) and platinum (Pt) was carried out using ICP-MS in 34 samples of wild fungi and 48 samples of wild blackberries collected from sites across the UK. On a fresh weight basis (mg/kg) levels of Pb were in the range 0.003-5.990, Cu 0.596-34.800, Cd<0.001-19.6, Hg<0.001-4.150, As 0.001-0.972 and Pt (microg/kg) 0.006-0.200, with higher concentrations found in fungi than in blackberries. The results of the survey showed that the concentrations of the metals were consistent with previous studies, where available. Concentrations in wild fungi of Pt, tin (Sn), and titanium (Ti) were significantly higher at urban sites than at rural sites. Urban blackberries had significantly higher levels of Pb, Ti, and Cd than rural ones, but lower levels of manganese (Mn). Pb, Ti and Sn concentrations were significantly higher in blackberries sampled near main roads rather than in rural areas.

Environmental Monitoring↗
PubMed2001

The prevalence of Chlamydia trachomatis infection in male undergraduates: a postal survey.

OBJECTIVES: To determine the prevalence of Chlamydia trachomatis infection in male undergraduates and to investigate whether prevalence increases with time spent at university. To investigate the feasibility of screening men for C trachomatis by self sampling and posting of urine specimens. METHODS: The study design was a postal survey undertaken by the Department of Genito-Urinary Medicine (GUM) and Student University Health Service (SUHS) in SHEFFIELD: 2607 male undergraduates from the SUHS patient list were invited to participate in the study by providing a first void urine specimen and posting it to the laboratory. The main outcome measure was the detection of C trachomatis infection. RESULTS: 758 students participated in the study, a response rate of 29.1%. Nine students (1.2%) tested positive for C trachomatis. The prevalence of infection in the first, second, and third year of study was 0.7%, 1.5%, and 1.6% of participants respectively. There was no statistically significant difference in prevalence of infection between first and third year students (chi(2) test, p = 0.32). However, students with chlamydia had a higher median age (Mann-Whitney U test, p < or = 0.05). Contact tracing identified four further cases of C trachomatis infection. CONCLUSION: Screening for C trachomatis infection by postal survey is feasible. However, the response rate in this study was poor and the estimated sample size was not reached. Therefore, it has not been possible to determine the true prevalence of infection in this population or to accurately assess changes in prevalence with time spent at university.

Adolescent↗
PubMed2000

Single-molecule analysis of DNA immobilized on microspheres.

The formation and analysis of single molecules of fluorescently labeled DNA immobilized on polystyrene microspheres is described. Analysis by confocal fluorescence microscopy revealed single-step photobleaching, characteristic of a single fluorophore. Microspheres provide a means of locating single molecules by bright-field microscopy, prior to single-molecule detection. This allows the interrogation of single molecules without suffering the limitations of premature photobleaching. Statistical analysis of fluorescence intensities for >100 microspheres suggests attachment of DNA to micropsheres to be consistent with Poisson statistics.

DNA↗
PubMed2000

Acclimatizing the world: a history of the paradigmatic colonial science.

This paper examines the institutions, personages, and the theories that informed acclimatization activities in nineteenth-century France, England, and the two colonies of Algeria and Australia. Treating acclimatization as a scientific concept and activity the essay begins with the conditions of its emergence in Enlightenment France. Subsequent sections trace the growth of the acclimatization movement and its translation to the British context, and consider reasons for its decline in the last third of the nineteenth century. Efforts are made to show why many perceived acclimatization to be the paradigmatic colonial science with applications as diverse as agriculture, settlement schemes, field sports, and human health. Emphasis falls on the French and British cultural spheres, as these were the dual epicenters of both modern colonialism and organized acclimatization activity.

Acclimatization↗
PubMed2000

Groucho/TLE/R-esp proteins associate with the nuclear matrix and repress RUNX (CBF(alpha)/AML/PEBP2(alpha)) dependent activation of tissue-specific gene transcription.

The Runt related transcription factors RUNX (AML/CBF(alpha)/PEBP2(alpha)) are key regulators of hematopoiesis and osteogenesis. Using co-transfection experiments with four natural promoters, including those of the osteocalcin (OC), multi drug resistance (MDR), Rous Sarcoma Virus long terminal repeat (LTR), and bone sialoprotein (BSP) genes, we show that each of these promoters responds differently to the forced expression of RUNX proteins. However, the three RUNX subtypes (i.e. AML1, AML2, and AML3) regulate each promoter in a similar manner. Although the OC promoter is activated in a C terminus dependent manner, the MDR, LTR and BSP promoters are repressed by three distinct mechanisms, either independent of or involving the AML C terminus, or requiring only the conserved C-terminal pentapeptide VWRPY. Using yeast two hybrid assays we find that the C terminus of AML1 interacts with a Groucho/TLE/R-esp repressor protein. Co-expression assays reveal that TLE proteins repress AML dependent activation of OC gene transcription. Western and northern blot analyses suggest that TLE expression is regulated reciprocally with the levels of OC gene expression during osteoblast differentiation. Digital immunofluorescence microscopy results show that TLE1 and TLE2 are both associated with the nuclear matrix, and that a significant subset of each colocalizes with AML transcription factors. This co-localization of TLE and AML proteins is lost upon removing the C terminus of AML family members. Our findings indicate that suppression of AML-dependent gene activation by TLE proteins involves functional interactions with the C terminus of AML at the nuclear matrix in situ. Our data are consistent with the concept that the C termini of AML proteins support activation or repression of cell-type specific genes depending on the regulatory organization of the target promoter and subnuclear localization.

Animals↗
PubMed1999

Cardiorespiratory dynamics: sensitivity of the on-transition to endurance-training status.

This project investigated the sensitivity of oxygen uptake (VO(2)) dynamics to training-induced physiological changes, across a broad spectrum of endurance-training histories. Forty subjects participated: sedentary (n = 10), active healthy (n = 10), regular runners (n = 10), and competitive distance runners (n = 10). Subjects completed a cycle step-function protocol, to elicit a steady state at 60% maximal work rate. Breath-by-breath data were collected for VO(2) and cardiac frequency (f(c)), and modelled mathematically, and used to determine the average response times to attain 20%, 40%, 60%, 80% and 100% of the respective steady states. The between-group comparisons for both VO(2) and f(c) revealed significantly faster response times to 40%, 60%, 80% and 100% of the induced response, for the better trained subjects (P < 0.05). In general, this technique permitted differentiation between the VO(2) and f(c) response dynamics of non-elite subjects from a broad range of endurance-training histories, with differences becoming more pronounced as subjects approached the steady state.

Adolescent↗
PubMed1998

Use of fluorescence resonance energy transfer to investigate the conformation of DNA substrates bound to the Klenow fragment.

Fluorescence resonance energy transfer (FRET) has been used to investigate the conformation of the single stranded region for a series of fluorescent DNA template-primers bound to the Klenow fragment (KF) of Escherichia coli DNA polymerase I. Fluorescent derivatives of template-primer DNA, modified with tetramethylrhodamine (TMR), served as energy transfer acceptors to the donor fluorescein fluorophore used to modify cysteine 751 in the double mutant KF (S751C, C907S). Design of the template-primer allowed the probe's position within the DNA-protein complex to be varied by stepwise extension of the primer strand upon addition of the appropriate deoxynucleoside triphosphates (dNTP). The TMR acceptor probe occupied seven different positions in the template-primers, five in the single stranded region and two in the double stranded region. The efficiency of energy transfer was determined at each position by calculating the integrated area of the fluorescein emission peak in the presence and absence of acceptor. Results indicate that the FRET efficiency varied in a sinusoidal fashion with a periodicity of approximately 10 base pairs and that the data could be fitted to an equation derived from a simple model formulated on the basis of helical structure. The data support the conclusion that the single stranded template portion of a DNA template-primer adopts a helical conformation when bound to the KF. The results of this study further support FRET as a useful method for the determination of structure and conformation in protein-DNA complexes.

DNA Polymerase I↗
PubMed1997

Nonoperative estimation of the soleus musculotendinous junction using magnetic resonance imaging.

Although it is recognized as the muscle flap of choice for middle-third defects of the lower limb, the capability for even more distal transposition of the soleus muscle remains controversial. Such reach depends directly on the site of insertion of the muscle and previously has not been assessed convincingly without surgical intervention. Magnetic resonance imaging (MRI) may be a noninvasive alternative for determining the distal extent of the musculotendinous junction of the soleus muscle. In our last four patients, preoperative MRI scans were obtained prior to an elective soleus muscle transfer. The distance from the ankle joint to the most distal site of the soleus insertion was measured on the MRI scan and compared with the actual intraoperative measurement, which had a significant correlation (r = 0.98, p = 0.019). A retrospective review of 42 other sagittal ankle MRI scans predicted the mean of this distance to be 1.92 +/- 1.23 cm (range -0.4 to 4.5 cm), compared with gross anatomic dissections in 30 unrelated fresh cadavers, where this was 4.06 +/- 3.11 cm (range -0.7 to 12.5 cm). These additional data are pertinent because they reinforce recognition of the great variation in soleus anatomy, which would limit clinical applications for the distal third of the leg only for those individuals with very distal insertions. The MRI scan can reliably identify the soleus muscle and provides a nonoperative method for evaluation of potential feasibility for its use as a local muscle flap for distal lower extremity defects.

Ankle↗
PubMed1996

The inositol 5'-phosphatase SHIP binds to immunoreceptor signaling motifs and responds to high affinity IgE receptor aggregation.

Immunoreceptors such as the high affinity IgE receptor, FcepsilonRI, and T-cell receptor-associated proteins share a common motif, the immunoreceptor tyrosine-based activation motif (ITAM). We used the yeast tribrid system to identify downstream effectors of the phosphorylated FcepsilonRI ITAM-containing subunits beta and gamma. One novel cDNA was isolated that encodes a protein that is phosphorylated on tyrosine, contains a Src-homology 2 (SH2) domain, inositolpolyphosphate 5-phosphatase activity, three NXXY motifs, several proline-rich regions, and is called SHIP. Mutation of the conserved tyrosine or leucine residues within the FcepsilonRI beta or gamma ITAMs eliminates SHIP binding and indicates that the SHIP-ITAM interaction is specific. SHIP also binds to ITAMs from the CD3 complex and T cell receptor zeta chain in vitro. SHIP protein possesses both phosphatidylinositol-3,4,5-trisphosphate 5'-phosphatase and inositol-1,3,4,5-tetrakisphosphate 5'-phosphatase activity. Phosphorylation of SHIP by a protein-tyrosine kinase, Lck, results in a reduction in enzyme activity. FcepsilonRI activation induces the association of several tyrosine phosphoproteins with SHIP. SHIP is constitutively tyrosine-phosphorylated and associated with Shc and Grb2. These data suggest that SHIP may serve as a multifunctional linker protein in receptor activation.

Animals↗
PubMed1996

Inorganic fluoride concentration after long-term sedation with isoflurane.

We report on five patients in whom long-term sedation with isoflurane for up to 7 days was used successfully. Serum inorganic fluoride concentrations were measured daily. The concentrations ranged from 12 mmol l-1 to 29 mmol l-1. These were well below the described renal toxic level of 50 mmol l-1. There was no deterioration in renal function attributable to the use of isoflurane.

Adult↗
PubMed1996

Role of magnetic resonance imaging in the diagnosis of osteomyelitis in diabetic foot infections.

PURPOSE: The role of magnetic resonance imaging (MRI) in the diagnosis of osteomyelitis in foot infections in diabetics was investigated. The accuracy, sensitivity, and specificity of MRI, plain radiography, and nuclear scanning were determined for diagnosing osteomyelitis, and a cost comparison was made. METHODS: Twenty-seven patients with diabetic foot infections were studied prospectively. All patients underwent MRI and plain radiography. Twenty-two patients had technetium bone scans, and 19 patients had Indium scans. Nineteen patients had all four tests performed. Patients with obvious gangrene or a fetid foot were excluded. RESULTS: The diagnosis of osteomyelitis was established by pathologic specimen (n = 18), bone culture (n = 3), or successful response to medical management (n = 6). Osteomyelitis was confirmed in nine of the pathologic specimens. The diagnostic sensitivity, specificity, and accuracy for MRI was 88%, 100%, and 95%, respectively, for plain radiography it was 22%, 94%, and 70%, respectively, for technetium bone scanning it was 50%, 50%, and 50%, respectively, and for Indium leukocyte scanning it was 33%, 69%, and 58%, respectively. The data were analyzed statistically with the two-tailed Fisher's exact test. MRI was the only test that was statistically significant (p < 0.01). CONCLUSIONS: MRI appeared to be the single best test for the diagnosis of osteomyelitis associated with diabetic foot infections. It had a better diagnostic accuracy than conventional modalities and appeared to be more cost-effective than the frequently used Indium scan.

Adult↗
PubMed1995

Carotid artery stenosis: preoperative noninvasive evaluation in a community hospital.

PURPOSE: The purpose of this study was to determine whether noninvasive evaluation with duplex ultrasonography and magnetic resonance angiography of patients with carotid artery stenosis can replace contrast angiography at our institution. METHODS: This study consisted of a retrospective chart review of 40 patients (74 carotid arteries) in combination with a blinded reanalysis of original data. Contrast angiography was compared with duplex ultrasonography and magnetic resonance angiography. The overall diagnostic accuracy of duplex ultrasonography and magnetic resonance angiography was determined individually and concordantly in patients being evaluated for carotid artery stenosis. RESULTS: The overall sensitivity of duplex ultrasonography was 88.5%, and the specificity was 91.7% (Spearman correlation coefficient = 0.8456; p < 0.001). For magnetic resonance angiography the sensitivity was 92.3%, and the specificity was 97.9% (Spearman correlation coefficient = 0.9086; p < 0.001). In the presence of concordance, the noninvasive studies exhibited a sensitivity of 100%, (correlation coefficient = 0.9661; kappa value = 0.9655). No occlusions or severe lesions were missed by both studies. In only one vessel (1.52%) was a false-positive concordance noted. CONCLUSIONS: Carotid endarterectomy may be undertaken with a high degree of confidence that the operation will be appropriate if the noninvasive evaluations are concordant. In the absence of concordance of the noninvasive studies, contrast angiography should be considered.

Aged↗
PubMed1995

The yeast tribrid system--genetic detection of trans-phosphorylated ITAM-SH2-interactions.

Protein-protein interactions are often dependent on the post-translational modification of one component of a complex. To facilitate the study of these interactions in signal transduction, we have developed the yeast tribrid system, a modification of the yeast two-hybrid system. We demonstrate that the interactions are dependent upon the presence of a tyrosine kinase, an SH2 domain and a tyrosine containing substrate. Using the gamma subunit of the high-affinity IgE receptor, Fc epsilon RI, this approach has been used to isolate a novel SH2-containing family member. The mRNA encoding this novel protein is differentially expressed in rat tissues. The yeast tribrid system can be readily adapted for the characterization of novel tyrosine kinases or substrates, as well as the study of protein-protein interactions which involve other post-translational modifications.

Adaptor Proteins, Signal Transducing↗
PubMed1994

Nuf2, a spindle pole body-associated protein required for nuclear division in yeast.

The NUF2 gene of the yeast Saccharomyces cerevisiae encodes an essential 53-kd protein with a high content of potential coiled-coil structure similar to myosin. Nuf2 is associated with the spindle pole body (SPB) as determined by coimmunofluorescence with known SPB proteins. Nuf2 appears to be localized to the intranuclear region and is a candidate for a protein involved in SPB separation. The nuclear association of Nuf2 can be disrupted, in part, by 1 M salt but not by the detergent Triton X-100. All Nuf2 can be removed from nuclei by 8 M urea extraction. In this regard, Nuf2 is similar to other SPB-associated proteins including Nufl/SPC110, also a coiled-coil protein. Temperature-sensitive alleles of NUF2 were generated within the coiled-coil region of Nuf2 and such NUF2 mutant cells rapidly arrest after temperature shift with a single undivided or partially divided nucleus in the bud neck, a shortened mitotic spindle and their DNA fully replicated. In sum, Nuf2 is a protein associated with the SPB that is critical for nuclear division. Anti-Nuf2 antibodies also recognize a mammalian 73-kd protein and display centrosome staining of mammalian tissue culture cells suggesting the presence of a protein with similar function.

Amino Acid Sequence↗