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Biomedical subjects

G Murphy

Publications and source records attributed to G Murphy.

At least 361 records · Page 20Linked to original sources
PubMed1988

Degradation of glomerular basement membrane by purified mammalian metalloproteinases.

Neutral metalloproteinases degrade components of the extracellular matrix, including collagen types I-V, fibronectin, laminin and proteoglycan. However, their ability to degrade intact glomerular basement membrane (GBM) has not previously been investigated. Incubation of [3H]GBM (50,000 c.p.m.; pH 7.5; 24 h at 37 degrees C) with purified gelatinase or stromelysin (2 units) resulted in significant GBM degradation: gelatinase, 46 +/- 2.2; stromelysin, 59 +/- 5.8 (means +/- S.E.M.; percentage release of non-sedimentable radioactivity; n = 4). In contrast, 2 units of collagenase released only 5.6 +/- 0.52% (n = 3) of the [3H]GBM radioactivity compared with 2.0 +/- 0.15% (n = 7) released from [3H]GBM incubated alone. Sephadex G-200 gel chromatography of supernatants obtained from incubations of [3H]GBM with either gelatinase or stromelysin confirmed the ability of these enzymes to degrade GBM and revealed both high-(800,000) and relatively low-(less than 20,000) Mr degradation products for both enzymes. GBM degradation by gelatinase and stromelysin was dose-dependent (range 0.02-2.0 units), near maximal between pH 6.0 and 8.6, and was completely inhibited (greater than 95%) by 2 mM-o-phenanthroline. Collagenase (2 units) did not enhance the degradation of GBM by either gelatinase (0.02 or 0.2 unit) or stromelysin (0.02 or 0.2 unit). Our results indicate that metalloproteinase-mediated GBM degradation by neutrophils and glomeruli may be attributable to gelatinase (neutrophils) and/or stromelysin (glomeruli) and suggest an important role for these proteinases in glomerular pathophysiology.

Animals↗
PubMed1988

Plasma protein analysis by immunoblotting.

We have demonstrated the value of immunoblotting techniques for the examination of the variant forms of plasma proteins. Furthermore, by preincubation of samples with the appropriate enzymes or ligands it is possible, using immunoblotting, to examine the molecular basis and functional significance of plasma protein microheterogeneity.

Blood Proteins↗
PubMed1988

Pulsed dye laser (577 nm) treatment of portwine stains: ultrastructural evidence of neovascularization and mast cell degranulation in healed lesions.

Portwine stains were examined before, immediately after, and 1 yr after successful clearance by a pulsed dye laser (577 nm) using ultrastructural techniques. Dilated vascular channels and mast cell hypoplasia characterized lesional skin before treatment. Immediately after treatment, widespread selective vessel necrosis, similar to changes previously described, was observed. One year after laser irradiation, the abnormally ectatic portwine stain vessels had been replaced by small venules and arterioles, similar in number and diameter to blood vessels in normal skin; the only difference noted was that these new vessels were surrounded by easily identifiable mast cells. Many of these mast cells exhibited evidence of activation and degranulation. We conclude that mast cells may play an important role in the neovascularization of portwine stains treated by 577-nm dye laser irradiation.

Adolescent↗
PubMed1988

Effects of supplemental Bermuda grass hay or corn on intake, digestion and performance of cattle consuming endophyte-infected fescue.

Effects of supplemental Bermuda grass hay (BG) or ground corn on intake, digestion and performance of cattle consuming endophyte-infected fescue (I) were studied. In Exp. 1, a Latin square study, five growing Holstein steers (158.1 kg) consumed I ad libitum and were offered 0, .3, .6, .9 or 1.2% body weight (BW) of BG daily. Total dry matter (DM) intake rose linearly (P less than .05) with increasing BG, although intake was numerically similar with .6, .9 and 1.2% BW of BG. Digestibility was constant with diet (P greater than .10). Six growing Holstein steers used in Exp. 2, a Latin square with a 2 x 3 factorial arrangement of treatments, ingested I or noninfected (NI) fescue hay ad libitum with 0, .5 or 1.0% BW of ground corn. Total DM intake increased linearly as the level of corn rose (P less than .05). Total intake with I increased more with the first than with the second addition of corn, and the opposite occurred with NI (interaction between fescue infection and the quadratic effect of corn level, P less than .10). Organic matter digested (g/d) was greater for NI than for I and rose linearly with increasing corn ingestion (P less than .05). Ninety-six crossbred beef heifers and steers (184.2 kg avg initial live weight) were used in a 77-d fall grazing experiment (Exp. 3) with a 2 x 3 factorial treatment arrangement. Cattle grazed I or NI paddocks and were given no supplement or .34% BW of BG or .65% BW of ground corn on a daily basis (DM).(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗
PubMed1988

Pain in peripheral arteriography: an assessment of conventional versus ionic and non-ionic low-osmolality contrast agents.

Two low-osmolality contrast agents, ioxaglate meglumine/sodium and iohexol were compared with diatrizoate meglumine/sodium in a controlled double blind study of 126 patients undergoing arteriography for peripheral vascular disease to determine which caused the least pain. Discomfort was assessed by means of a visual analog scale rating pain from 0 to 100. Average values for pain were 39 +/- 27 for diatrizoate, 14 +/- 15 for ioxaglate and 21 +/- 22 for iohexol. We found that both low-osmolality agents caused significantly less pain in peripheral arteriography than the traditional agent. The p values were p less than 0.0005 for ioxaglate and p less than 0.005 for iohexol versus diatrizoate. In addition, ioxaglate was found to cause significantly less pain than iohexol (p less than 0.05) in this patient group.

Adult↗
PubMed1987

Stromelysin is an activator of procollagenase. A study with natural and recombinant enzymes.

The latent forms of stromelysin and collagenase from human gingival fibroblasts were purified to homogeneity. These latent proenzymes underwent serial small reductions in Mr upon activation by treatment with either 4-aminophenylmercuric acetate or trypsin. Similar shifts in Mr and activation kinetics were observed upon identical treatments of either recombinant prostromelysin or procollagenase. Prostromelysin showed a lag between activation and Mr fall, suggesting an initial activation by conformational change. Collagenase activity was enhanced up to 12-fold by either natural or recombinant stromelysin in the presence of trypsin or 4-aminophenylmercuric acetate. Stromelysin caused a further apparent decrease in the Mr of procollagenase. Since these important connective-tissue-degrading enzymes are usually co-ordinately produced by cells, a cascade mechanism is proposed in which collagenase is activated by stromelysin.

Collagenases↗
PubMed1987

Transforming growth factor beta modulates the expression of collagenase and metalloproteinase inhibitor.

Exposure of quiescent MRC-5 human fibroblasts to growth factors such as epidermal growth factor, basic fibroblast growth factor or embryonal carcinoma-derived growth factor resulted in the induction of mRNA transcripts encoding the metalloproteinases collagenase and stromelysin and the specific metalloproteinase inhibitor TIMP, whilst expression of collagen and fibronectin was relatively unaffected. Exposure of quiescent cells to growth factors in the presence of transforming growth factor beta (TGF-beta) resulted in inhibition of collagenase induction and a synergistic increase in TIMP expression. TGF-beta alone did not significantly induce metalloproteinase or TIMP expression. These effects on mRNA transcripts were reflected in increased secretion of TIMP protein and collagenase activity. Nuclear run-off analysis of growth factor-induced transcription revealed that the TGF-beta modulation of TIMP and collagenase expression was due to transcriptional mechanisms. The observations suggest that TGF-beta exerts a selective effect on extracellular matrix deposition by modulating the action of other growth factors on metalloproteinase and TIMP expression.

Cell Line↗
PubMed1987

Tissue inhibitor of metalloproteinases (TIMP) regulates extracellular type I collagen degradation by chondrocytes and endothelial cells.

A specific antiserum to purified rabbit tissue inhibitor of metalloproteinases (TIMP) was raised in sheep, characterized and used to investigate the role of TIMP in a model system. Chondrocytes and endothelial cells cultured on 14C-labelled type I collagen films and stimulated to produce collagenase were unable to degrade the films unless the anti-TIMP antibody was added. The degradation induced was inhibited by a specific anti-rabbit collagenase antibody. It was concluded that TIMP is a major regulatory factor in cell-mediated collagen degradation.

Animals↗
PubMed1986

Comparison of human stromelysin and collagenase by cloning and sequence analysis.

A comparison of the cDNA-derived amino acid sequences of human stromelysin and collagenase with the N-terminal sequences of purified enzymes reveals that these metalloproteinases are highly conserved and that they are secreted as proenzymes. A putative zinc-binding site was identified by its homology with the zinc-chelating sequence of thermolysin. These sequences permitted the identification of: transin, a protein induced in rat fibroblasts either exposed to growth factors or transformed by oncogenic viruses, as the rat homologue of stromelysin, and XHF1, a protein induced in human fibroblasts after treatment with tumourigenic agents, as collagenase.

Amino Acid Sequence↗
PubMed1986

Rapid purification of tissue inhibitor of metalloproteinases from human plasma and identification as a gamma-serum protein.

A rapid method is described for the purification of human tissue inhibitor of metalloproteinases (TIMP) from plasma which involves immuno-affinity chromatography and gel filtration. The purified plasma inhibitor is immunologically identical with the TIMP previously purified from human amniotic fluid, human synovial fluid and human fibroblast culture medium. It is proposed that this inhibitor is identical with the plasma inhibitor previously named 'B1 anticollagenase', although the plasma inhibitor was shown to migrate as a gamma-serum component.

Blood Proteins↗