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Biomedical subjects

D Wilkinson

Publications and source records attributed to D Wilkinson.

At least 289 records · Page 16Linked to original sources
PubMed1989

Intercellular adhesion molecule 1 is induced on isolated endocrine islet cells by cytokines but not by reovirus infection.

The selective destruction of the pancreatic islet beta cells in type 1 diabetes mellitus is thought to be mediated by a cellular autoimmune process, possibly triggered by virus infection in genetically susceptible individuals. Because of the potentially important role of cell-cell adhesion in the immune response, we investigated whether cytokine products of mononuclear cells, or virus infection, induced the expression of intercellular adhesion molecule 1 (ICAM-1) on human endocrine islet cells. By flow cytofluorimetry, control islet cells did not express detectable ICAM-1. However, after a 72-hr exposure of islets to interferon gamma (IFN-gamma) and/or tumor necrosis factor alpha (TNF-alpha) (each at 250 units/ml), ICAM-1 was induced on greater than 85% of islet cells. IFN-gamma was 50% more potent than TNF-alpha; together, their effects were additive. Class I major histocompatibility complex (MHC) protein expression, detected on control islet cells, was also stimulated by IFN-gamma and/or TNF-alpha. In contrast, infection with reovirus type 3 did not induce ICAM-1 on islet cells, although it stimulated the expression of class I MHC proteins. By double-label indirect immunofluorescence microscopy, ICAM-1 expression was identified on both beta (insulin-secreting) and delta (somatostatin-secreting) islet cells. Monoclonal antibody to ICAM-1 precipitated protein of Mr 97,000 from [35S]methionine-labeled islets exposed to IFN-gamma and TNF-alpha, but not from control islets. RNA blot analysis revealed a major species of 3.3 kilobases and a minor species of 2.2 kilobases induced in islets exposed to the cytokines. These findings have implications for the molecular mechanisms of beta-cell destruction in type 1 diabetes, in that expression of ICAM-1 by beta cells may facilitate adhesion of antigen-targeted immune cells.

Antibodies, Monoclonal↗
PubMed1989

The radiation dose to cells in vitro from intracellular indium-111.

Most of the radionuclides used in nuclear medicine emit low energy Auger electrons following radioactive decay. These emissions, if intracellular, could irreparably damage the radiosensitive structures of the cell. The resulting radiation dose, which is a measure of biological damage in the affected cell, could be many times the average radiation dose to the associated organ. In this series of experiments, the radiation dose to the nucleus of a chinese hamster V79 cell was determined for the intracellular radiopharmaceutical 111indium-oxine. Assuming the cell nucleus to be the radiosensitive volume, the radiation dose would be primarily due to the low energy Auger electrons. A much smaller dose would be absorbed from the penetrating X- and gamma-rays and internal conversion electrons released from other radiolabelled cells in the culture. The radiation dose to the cell from the intranuclear decay of 111In was empirically established from cell survival studies to be 3.5 mGy/decay, using cobalt-60 as a reference radiation. The average dose to V79 cells from extracellular 111In (i.e., from 111In located outside the target cell) was calculated to be 5.8 pGy/decay. This suggests that for an intracellular radiopharmaceutical, the radiation dose of consequence would be delivered by the low energy Auger electrons. In contrast, Auger electrons from an extracellular radiopharmaceutical could not directly damage the cell nucleus and therefore would not contribute to the radiation dose.

Animals↗
PubMed1989

Hip joint mobility and spinal angles in standing and in different sitting postures.

The spinal angles of 25 female subjects were measured in standing and in four different sitting postures with various trunk-thigh angles. Additionally, measurements of hip and lumbar mobility were made in an attempt to relate mobility to spinal curvature in the different sitting postures. Lumbar curvature was observed to decrease as the trunk-thigh angle decreased across the different sitting postures, and reduction in curvature was associated with hip mobility. An exploratory analysis of interrelationships among hip mobility, spinal mobility, and spinal curvature was also carried out. A significant correlation between lumbar and thoracic angular deviations in standing and hip flexion/extension range was obtained. The findings are discussed with reference to theories of sitting posture and the influence of hip joint mobility on postural adaptations to furniture. Further investigations, including studies of male subjects, are indicated.

Adult↗
PubMed1989

Controlled trial of cephradine versus cefuroxime in vascular surgery.

Two hundred and three consecutive patients undergoing acute or elective vascular reconstructions (N = 162) or amputations (N = 41) were randomized to receive either a single dose of cephradine 2 g intravenously or cefuroxime 1.5 g intravenously at induction of anaesthesia. Infective morbidity in both groups was assessed post-operatively as was therapeutic antibiotic prescribing. No significant differences in septic complications were found between patients receiving cefuroxime or cephradine. In addition, tissue penetration of each antibiotic was assessed by assay of serum and tissue specimens. Serum levels of cefuroxime were significantly less than cephradine 10 min after injection (median concentrations 115 micrograms/ml versus 182 micrograms/ml, p less than 0.01 Wilcoxon), but there were no differences in tissue penetration.

Amputation, Surgical↗
PubMed1989

The clinical application of forearm and hand blood flow measurements using a gamma camera.

A new method of measuring blood flow to forearms and hands during reactive hyperemia is described. Subjects are positioned with forearms and hands over a gamma camera, and the circulation below the elbows is isolated for 4 minutes with pneumatic cuffs. The remaining blood pool is labeled with technetium-99m-labeled human serum albumin given via a vein in the dorsum of the foot. The rate of rise of activity in the arms after cuff release is measured from the gradient of time activity curves, and these are calibrated for flow by counting a sample of venous blood. Blood flow per unit volume is obtained from measurements of the limb volume, done by water displacement. Results for eight normal control subjects showed a mean flow per unit volume to forearm plus hand of 32.9 +/- 6.4 ml/100 ml/min and to hand alone 36.5 +/- 6.9 ml/100 ml/min. Case studies of patients with a variety of large- and small-vessel diseases are presented, illustrating the use of the technique to aid diagnosis and measure response to treatment.

Adult↗
PubMed1989

Forearm blood flow measurements using technetium-99m human serum albumin following brachial arteriotomy.

A simple, low cost method for measuring forearm blood flow during reactive hyperemia has been developed. Subjects are seated with hands and forearms over a large field-of-view gamma camera. Blood pressure cuffs inflated above the elbows isolate the blood in the forearms and hands and induce a hyperemic response. The remaining blood pool is labeled with technetium. The rate of increase of activity following release of the cuffs is measured from the gradient of time-activity curves and is calibrated for flow by counting a venous blood sample. The technique has been applied to a group of normal controls and to symptomatic and asymptomatic patients following right brachial arteriotomy. Forearm blood flow in normal subjects was 32.9 +/- 6.4 ml/100 ml/min and for subjects with occlusion of the brachial artery was 6.4 +/- 2.1 ml/100 ml/min. The method is simple, widely available, and reproducible. The good signal to noise ratio allows it to be used in cases of very low flow either as an aid to diagnosis or to measure treatment response.

Adult↗
PubMed1988

Analysis of HLA-DR glycoproteins by DNA-mediated gene transfer. Definition of DR2 beta gene products and antigen presentation to T cell clones from leprosy patients.

We have used DNA-mediated gene transfer to express HLA class II molecules in mouse L cells for serological, biochemical, and functional analysis. cDNA clones encoding the DR2 beta a and DR2 beta b products of the DR2Dw2 haplotype were subcloned into a mouse Moloney leukemia virus-based expression vector (pJ4) and transfected separately into mouse L cells together with a HLA-DR alpha/pJ4 construct. These transfectants have allowed differential analysis of the two DR2 beta products in a manner normally prohibited by the concomitant expression seen in B cells. Two-dimensional SDS-PAGE analysis of the transfectants defines the more acidic beta chain as the product of the DR2 beta a sequence, and the more basic chain as the product of the DR2 beta b sequence. The LDR2a transfectants present antigen efficiently to M.leprae-specific T cell clones and are capable of presenting synthetic peptide, 65-kD recombinant mycobacterial antigen and M.leprae. Of the DR2Dw2-restricted T cell clones we have tested, all use the DR2 beta a chain as their restriction element. Inhibition studies with mAbs demonstrate the dependence of presentation by the transfectant on class II and CD4, while mAbs against LFA-1, which substantially inhibit presentation by B-lymphoblastoid cell lines, do not inhibit transfectant presentation.

Amino Acid Sequence↗
PubMed1988

Prediction and identification of an HLA-DR-restricted T cell determinant in the 19-kDa protein of Mycobacterium tuberculosis.

An allele-specific motif has been identified in the sequence of several peptides which are recognized by T cells in association with HLA-DR1. In order to test the predictive values of such a motif we analyzed the 19-kDa antigen from Mycobacterium tuberculosis and identified a sequence containing a pattern characteristic of DR1 restriction. Peripheral blood mononuclear leukocytes from every DR1 and 4 individual tested responded to the corresponding synthetic peptide. Nine other donors, constituting seven different DR alleles, failed to recognize this sequence. Recognition of the peptide in association with DR1 and DR4 was confirmed using T cell clones and transfected murine L cell lines expressing DR molecules.

Amino Acid Sequence↗
PubMed1987

A reliable and readily available method of measuring limb blood flow in intermittent claudication.

To evaluate isotope limb blood flow measurement in intermittent claudication we have assessed 58 non-diabetic patients comparing our new method with treadmill testing and Doppler assessment. Limb blood flow was applicable to all 58 patients; 25 patients were unable to walk on a treadmill and of the 33 who could 12 failed to walk for one minute, making a standard one minute exercise test inappropriate. In those patients who could perform exercise tests there was a significant correlation between maximum walking distance and limb blood flow (r = 0.35, P = 0.02). Resting, post-exercise and post-hyperaemic ankle-brachial systolic pressure indices bore no relationship to the maximum walking distance. Isotope limb blood flow measurement is reproducible (r = 0.97), can be applied to those patients who cannot walk on a treadmill and provides information about both legs. It correlated significantly with all the other tests and can be recommended for the minimally invasive assessment of intermittent claudication.

Adult↗
PubMed1987

Sequence and evolution of HLA-DR7- and -DRw53-associated beta-chain genes.

cDNA clones representing products of the DR7 and DRw53 beta-chain genes were isolated from the human B-lymphoblastoid cell line MANN (DR7,DRw53,DQw2, DPw2). The DRw53 beta sequence was identical to a DRw53 beta sequence derived from cells with a DR4 haplotype. In contrast, the DR7 beta sequence was as unrelated to DR4 beta sequence as it was to other DR beta-related genes, except at the 3'-untranslated region. These results suggest that the DR7 and DR4 haplotypes may have been derived relatively recently from a common ancestral haplotype and that the DR4 and DR7 beta-chain genes have undergone more rapid diversification in their beta 1 domains, most probably as a result of natural selection, than have the DRw53 beta-chain genes. Short tracts of sequence within the DR7 and DRw53 beta 1 domains were shared with other DR beta sequences, indicating that exchanges of genetic information between beta 1 domains of DR beta-related genes have played a part in their evolution. Serological analysis of mouse L-cell transfectants expressing surface HLA-DR7 molecules, confirmed by antibody binding and allelic sequence comparisons, identified amino acid residues that may be critical to the binding of a monomorphic DR- and DP-specific monoclonal antibody.

Amino Acid Sequence↗
PubMed1987

Molecular cloning and primary structure of myelin-associated glycoprotein.

Myelin-associated glycoprotein (MAG) may play a role in the cellular interactions leading to myelination. Using monoclonal antibodies and conventional antisera against MAG, we have isolated a cDNA clone from an expression library prepared from rat brain mRNA. The identity of the clone was confirmed by the exact match between its nucleotide sequence and two peptide sequences of 13 and 9 amino acids that we obtained by Edman degradation of two CNBr fragments of MAG. The cDNA clone hybridized to two size species of mRNA in rat approximately 3.5 kilobases in length. These mRNAs were present in brain but not liver and were expressed most abundantly at the time of active myelination (day 14). The mRNA for MAG was present at barely detectable levels in hypomyelinating jimpy mice compared to normal littermate controls. Therefore the MAG cDNA clone is both brain and myelin specific. DNA sequence analysis revealed that our MAG cDNA was derived from the same mRNA as clone p1B236, a randomly selected, brain-specific, partial cDNA isolated by Sutcliffe et al. [Sutcliffe, J. G., Milner, R. J., Shinnick, T. M. & Bloom, F. E. (1983) Cell 33, 671-682]. Analysis of the predicted protein sequence suggests that MAG has a long extracellular domain (499 amino acids), followed by a short transmembrane segment (20 amino acids) and an intracellular carboxyl-terminal domain (90 amino acids). The molecule has several glycosylation sites, three internal repeats homologous to a repeat in the neural cell adhesion molecule (N-CAM), and sites for phosphorylation near the carboxyl terminus. The primary structure reported here provides a molecular framework for further investigations into the function of the MAG molecule.

Amino Acid Sequence↗