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Biomedical subjects

A Neil Turner

Publications and source records attributed to A Neil Turner.

5 recordsLinked to original sources
PubMed2004

Trace labeling of proteins with stable isotopes to identify fragments in complex mixtures.

We describe a novel nonradioactive protein-labeling technique that permits mass spectrometric identification of fragments of labeled proteins. Proteins are labeled by modulating their content of carbon-13 and labeled fragments identified from the distinctive isotope pattern observed on MALDI-TOF mass spectrometry. We show that carbon-13 enrichment to just 2.3% of total carbon (about twice the natural abundance of 1.1%) is sufficient for all fragments to be distinguishable from fragments of natural carbon-13-content proteins. Distinguishing labeled fragments is easily accomplished by visual inspection of spectra, but importantly, we show that labeled fragments can also be identified by computer analysis of spectra using novel parameters we have derived. The technique is demonstrated for identification of fragments of carbon-13-enriched glutathione transferase within a complex mixture of unlabeled peptides by visual and computer analysis of MALDI-TOF mass spectra, but it could be developed to mass spectrometrically identify and characterize fragments of labeled proteins recovered from biological systems.

Automation↗
PubMed2004

Teaching Web authoring: valuable skills, paperless courses.

The Internet has become a core resource in higher education but has predominantly been used by institutions to provide resources for teaching, and by students to find information, which they then present in more or less traditional ways. In medical practice, the Internet has advanced more slowly but its importance is advancing rapidly. Already, it is the single most important source of information for patients when they learn of a new disease. Little has so far been done to encourage physicians or students to provide information in this way. The authors explored the feasibility of combining such education with a simplified method for assessment of group projects. These were produced as websites, which were then assessed online by tutors and examiners. Feedback was strongly positive. Drawbacks were few, and often shared with other types of publication. The resources needed to set up such a course are significant but most higher education institutions already have expertise and resources for Internet publishing. The course becomes almost paperless. Students become practically familiar with the benefits and pitfalls of providing medical information on the Internet, and capable of doing so themselves.

Authorship↗
PubMed2004

Retransplantation in Alport post-transplant anti-GBM disease.

BACKGROUND: Post transplant anti-glomerular basement membrane (GBM) disease affects up to 5% of patients with Alport's syndrome. Defects in the COL4A5 gene are responsible for most cases, and alpha 5(IV)NC1 is the usual target for alloantibodies. Gene deletions are more commonly associated with this complication than are point mutations. The disease is severe in renal allografts and nearly always results in graft loss. METHODS: Three cases of retransplantation in Alport's syndrome are described here in detail. All cases were started on immunosuppressive therapy early in the course of their disease and one patient (case 2) received pre-emptive anti-T-cell therapy (Campath IH). Anti-GBM antibodies in these cases were investigated by standard anti-GBM enzyme-linked immunosorbent assay (ELISA), by indirect immunofluorescence, and by Western blotting using collagenase-digested human GBM and recombinant type IV collagen NC1 domains made in insect cells. RESULTS: All cases showed early antibody and complement fixation to human GBM. Target alloantibodies were to alpha 5(IV)NC1 domain predominantly. Cases two and three gave negative results on standard ELISA for anti-GBM antibodies. Pathologic examination revealed crescentic glomerulonephritis, which was rapid in onset in case 1, blunted and less aggressive in case 3, and case 2 developed segmental necrosis without crescent formation. Neutrophilic infiltrates were an early feature in all 3 cases. All cases are compared with a review of all retransplanted cases in the literature. CONCLUSION: Alport anti-GBM disease is a severe disease in retransplanted patients. Anti-T-cell therapy seemed to modify the pathologic findings but did not prevent graft loss. Longer term plasma exchange and mycophenolate mofetil may attenuate the illness, but in these cases did not prevent graft loss. Western blotting detected alloantibodies to alpha 5(IV) NC1 domain and is more sensitive and specific for this disease than standard ELISAs.

Adolescent↗
PubMed2003

Mass and composition matrix-assisted laser desorption ionization time of flight analysis enabling inference of the sequence of most peptides where the protein of origin is known.

Mass spectrometers combining matrix-assisted laser-desorption ionization (MALDI) and time-of-flight analysis are among the most widely used in peptide analysis. They excel at accurate mass determinations on complex samples but, compared with tandem instruments, have very limited capacity to determine amino acid sequence through daughter ion analysis. Here we have investigated the sequence information that can be inferred from the masses of peptides in the special circumstance in which the peptides are known to be sub-sequences of known parent sequences. We show how sequence can be inferred from the measured m/z of a peptide (mass analysis) and examine the parameters that influence the level of confidence that can be placed in "inferred sequences". We further describe how specific amino acid modifications can be used with MALDI-TOF analysis to obtain partial composition information and demonstrate that combined mass and composition (MAC) analysis enables the sequences of most peptide ions to be inferred with very high confidence.

Amino Acid Sequence↗