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Biomedical subjects

E B Casey

Publications and source records attributed to E B Casey.

At least 19 recordsLinked to original sources

Role of glycine 1 and lysine 2 in the glycation of bovine gamma B-crystallin. Site-directed mutagenesis of lysine to threonine.

To determine the role of Gly-1 and Lys-2 of bovine gamma B-crystallin in glycation and cross-linking, Lys-2 was changed to Thr by site-directed mutagenesis. A polymerase chain reaction was used to perform site-directed mutagenesis on the third codon (AAG-->ACG) of bovine gamma B-crystallin cDNA. The wild type and mutant cDNAs were cloned into pMON5743 and expressed in JM101 Escherichia coli cells, and the identity of gamma B-crystallin was confirmed by Western blotting after purification by cation exchange high performance liquid chromatography. Glycation of gamma B-crystallin by [14C]glucose was reduced significantly due to the mutation of Lys-2, supporting the view that Lys-2 is a major glycation site. Peptide mapping showed the presence of two major labeled peptides containing N-terminal sequences, and in the mutant these peptides had longer retention times and reduced radioactivity. Amino acid analysis, after glycation with [14C]glucose, revealed N-terminal glycine as the most predominant glycation site. Lys-2 was glycated slower than Gly-1 but faster than Lys-163. Glycation with DL-glyceraldehyde showed an important role for both Gly-1 and Lys-2 in the glycation-mediated gamma B-crystallin cross-linking.

Amino Acid Sequence↗

Leukocyte function-associated antigen 1 (LFA-1) and CD44 are signalling molecules for cytoskeleton-dependent morphological changes in activated T cells.

Signaling through the leukocyte function-associated antigen 1 (LFA-1) molecule has previously been shown to induce homotypic aggregation in T cells and to induce cytoskeletal changes in T lymphoma cells. In this study we describe the induction of a dendritic phenotype associated with cytoskeletal rearrangement in activated human peripheral blood T cells stimulated with monoclonal antibody SPV-L7 to LFA-1 alpha. Maximal expression of this phenotype required 72 h preactivation with phorbol myristate acetate and expression was abolished using the protein kinase C inhibitor staurosporine. Monoclonal antibody to CD18, the beta-chain of LFA-1, did not induce this phenotype. Monoclonal antibody MEM 83 to presumably a discrete epitope on LFA-1 alpha did not induce this phenotype but induced homotypic aggregation. However, a monoclonal antibody to CD44 induced a similar phenotype in activated lymphocytes. Induction of both homotypic in activated lymphocytes. Induction of both homotypic aggregation and the dendritic phenotype was abolished by preincubation with soluble intracellular adhesion molecule 1 (ICAM-1). Cytoskeletal inhibitors prevented the morphological changes in SPV-L7-activated lymphocytes. Preincubation with tyrosine kinase inhibitor, protein kinase C inhibitors, and inhibitors of new protein synthesis also prevented these morphological changes. These data suggest that discrete epitopes on LFA-1 alpha may be capable of inducing discrete signals either for homotypic aggregation or for a dendritic phenotype. As both LFA-1 and CD44 are involved in the migration of lymphocytes through high endothelial venules, these data could suggest that these molecules transduce signals resulting in cytoskeletal modification necessary for lymphocyte transmigration.

Alkaloids↗

Lymphatic obstruction in rheumatoid arthritis.

We describe four patients with rheumatoid arthritis and unilateral upper limb oedema. In all cases, qualitative lymphoscintigraphy showed lymphatic obstruction in the affected limb.

Adult↗

Expression of CD44 on rheumatoid synovial fluid lymphocytes.

OBJECTIVES: To investigate the involvement of the adhesion molecule CD44 in the homing of lymphocytes to synovial tissue, by examining the density of expression and molecular mass of CD44 on rheumatoid synovial fluid lymphocytes. METHODS: Twenty patients with rheumatoid arthritis were studied. Peripheral blood and synovial fluid lymphocytes were isolated by Ficoll-Hypaque sedimentation. CD44 expression was analysed by two colour flow cytometry of CD3 positive T lymphocytes with calculation of mean fluorescence intensity. Expression of activation markers M21C5, M2B3, interleukin (IL)-2 receptor and transferrin receptor was quantitated. In addition, CD44 molecular mass was examined by Western blot in six patients. RESULTS: CD44 expression was markedly increased on synovial fluid T lymphocytes of rheumatoid patients relative to peripheral blood lymphocytes from the same individuals. CD44 molecular mass on peripheral blood mononuclear cells was 88 kDa, but that on synovial fluid lymphocytes was only 83 kDa. CD44 expression correlated significantly with expression of activation markers M21C5, M2B3, and the IL-2 receptor. CONCLUSIONS: Alterations in density of expression or of the molecular mass of CD44 could contribute to local tissue injury, either directly by facilitating adhesion, or indirectly through effects on other adhesion molecules.

Arthritis, Rheumatoid↗

Eosinophilic fasciitis: a good response with conservative treatment.

A 52 year old man developed progressive painful swelling of both calves and difficulty walking. Physical examination showed asymmetrical localised swelling with induration and tenderness on palpation. Peripheral blood eosinophilia was noted. Biopsy of deep fascia and muscle showed typical features of eosinophilic fasciitis. He was treated with non-steroidal anti-inflammatory drugs and intensive physiotherapy. The clinical features had completely resolved six months later.

Anti-Inflammatory Agents, Non-Steroidal↗

Changes in levels of IgM RF and alpha 2 PAG correlate with increased disease activity in rheumatoid arthritis during the puerperium.

In this prospective study of 24 pregnant patients with rheumatoid arthritis, quiescent disease activity in 21 patients (88%) during gestation was followed by more active disease in the puerperium in 19 patients (79%). Increased disease activity was reflected in a deterioration in manual dexterity and this was found to correlate with a post-partum rise in IgM rheumatoid factor (IgM RF) (r = 0.86) and a post-partum decline in pregnancy associated alpha-2 glycoprotein (PAG) (r = 0.44). The increase in IgM RF also correlated with increased disease activity measured by a visual analogue scale (r = 0.44). Changes in IgA RF were not observed. These results suggest that PAG and IgM RF could contribute to the modulation and pathogenesis of rheumatoid arthritis during pregnancy.

Adult↗

Neurofilament expression in human T lymphocytes.

The expression of intermediate filaments in normal cells is mainly determined by their embryonal developmental origin. Flow cytometry using monoclonal antibody RT97 demonstrated that neurofilament was detectable in the human HuT 78 T-cell line and on resting T lymphocytes. Expression was greatly increased on lymphocytes activated for 3 days with phorbol ester. Western blotting confirmed the presence of the 200,000 MW form of neurofilament in T lymphocytes. Stimulation of peripheral blood T cells with phorbol myristate acetate (PMA) or with anti-CD3 monoclonal antibodies resulted in a marked increase in detection of phosphorylated neurofilament on Western blotting. Stimulation of HuT 78 cells with anti-LFA-1 resulted in redistribution of neurofilament from a perinuclear spheroid core into dendritic processes. These data indicate that T cells activated through the T-cell receptor associated complex express an intermediate filament usually associated with neurally derived cells. The finding that neurofilament expression and organization are regulated by T-cell surface molecules suggests a role for this intermediate filament in T-cell function.

Blotting, Western↗

Audit of full blood count monitoring in patients on longterm gold therapy for rheumatoid arthritis.

The use of injectable gold in the treatment of rheumatoid arthritis (RA) is hampered by the high incidence of adverse reactions of which myelotoxicity is potentially the most serious. Regular full blood count (FBC) monitoring prior to each injection, a practice which requires much time and effort, has never been fully evaluated. Of 154 patients who were started on gold and followed by the lifetable method for up to 10 years, five patients were withdrawn from treatment because of myelotoxicity, thrombocytopenia in all cases: the two most serious cases occurred early at 3 and 7 months after starting treatment; three mild cases occurred at 17, 37 and 60 months, were of slow onset and reversed spontaneously. Clearly, FBC monitoring is justifiable in the first two years of treatment when the incidence of adverse reaction is highest. Further investigation is required to justify this practice after two years of uncomplicated treatment.

Arthritis, Rheumatoid↗

Hepatic function in ankylosing spondylitis.

Values for alkaline phosphatase and gamma glutamyl transpeptidase (GGTP) and the prevalence of their elevation was significantly higher in 35 patients with ankylosing spondylitis (AS) than in 35 age and sex matched controls. The abnormal enzyme levels appeared to reflect a non-specific reaction to inflammation and could thus aid in assessment of disease status.

Adult↗

Recovery of conduction velocity distal to a compressive lesion.

Nerve conduction studies have been carried out pre- and postoperatively on 26 hands of patients with a carpal tunnel syndrome. When examined six to eight weeks postoperatively, there had been no significant change in conduction velocity or action potential amplitude in the fingers distal to the compression. Velocity had increased over the carpal tunnel segment. When the patients were seen between 12 and 18 weeks postoperatively, velocity and amplitude had increased in the fingers. The group with marked electrical abnormalities had improved more than the group in which digital amplitude was originally within the control range. The improvement occurred too early to be due to regeneration after relief of compression. It is suggested that either maturation of regenerating fibres was delayed by the compression or a reversible change had occurred in surviving fibres.

Action Potentials↗

Diabetic amyotrophy.

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Diabetes Complications↗

Diabetic amyotrophy: a follow-up study.

A clinical follow-up study of 12 patients with diabetic amyotrophy is reported. Re-examination after an interval indicated that improvement had occurred in all but one instance, and had been maintained over an average follow-up period of four and a half years. Improvement in the neurological syndrome appeared to follow improvement in diabetic control or institution of treatment in those whose diabetes had not previously been diagnosed.Seven patients made a good functional recovery, three no longer having any muscular weakness. Five showed significant residual disability.

Action Potentials↗

Digital nerve action potentials in healthy subjects, and in carpal tunnel and diabetic patients.

A technique is described for stimulating and recording from nerves in the finger using surface electrodes. A decrease in amplitude and velocity was found with increasing age. In control subjects the digital potential was approximately one and a half times larger than the potential recorded at the wrist. In patients with carpal tunnel syndrome there was some reduction in amplitude and velocity of the digital potential, but the changes were more marked at the wrist. In diabetic patients more uniform changes were found in the two segments. The technique was particularly useful in enabling conduction velocity to be calculated in the digital nerves when no potential could be recorded at the wrist.

Action Potentials↗