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A Gresham

Publications and source records attributed to A Gresham.

5 recordsLinked to original sources

Oxidative stress mediates synthesis of cytosolic phospholipase A2 after UVB injury.

UVB irradiation has previously been shown to significantly increase phospholipase activity and prostaglandin synthesis. Because UVB irradiation is a potent oxidative stress, the role of active oxygen species in regulating UV-induced cPLA2 synthesis and phosphorylation was examined. In the present study, irradiation produced a 3-fold increase in synthesis within 6 h following irradiation. Phosphorylation of cPLA2 was also increased to a similar extent. UVB-induced synthesis and phosphorylation of cPLA2 could be inhibited by pretreatment with the antioxidants 2,2,5,7,8-pentamethyl-6-hydroxychromane (50 microM) or N-acetylcysteine (10 mM). Treatment of unirradiated cultures with the potent oxidant tert-butyl hydroperoxide (500 microM) also increased cPLA2 synthesis and phosphorylation, suggesting that oxidative injury is an important regulator of cPLA2 synthesis. Increased synthesis of cPLA2 correlated well with increased [3H]arachidonic acid release, PGE2 synthesis and lipid peroxidation in epidermis after oxidant or UVB treatment. The results indicate that UVB-induced upregulation of cPLA2 synthesis is mediated by UVB-induced formation of free radicals.

Acetylcysteine

Increased synthesis of high-molecular-weight cPLA2 mediates early UV-induced PGE2 in human skin.

Ultraviolet light (UV) B-induced inflammation is characterized by dramatic increases in prostaglandin E2 (PGE2) synthesis due to enhanced arachidonate deacylation from the membrane. Therefore, the effect of UV on sythesis, mass, and distribution of the high-molecular-weight phospholipase A2 (cPLA2) in cultured human keratinocytes and human skin was studied. The 105-kDa cPLA2 was demonstrated to be the critical enzyme in UV-induced PGE2 synthesis and erythema in the first 6 h postirradiation. Immunoprecipitation of 35S-labeled protein showed cPLA2 synthesis increased three- to fourfold 6 h after irradiation. Immunoprecipitated 32P-labeled cPLA2 demonstrated phosphorylation of cPLA2 was concurrently induced, suggesting that UV also activates cPLA2. This increase in cPLA2 synthesis and activation also closely correlated with increased PGE2 synthesis and [3H]arachidonic acid release and was effectively blocked by both an S-oligonucleotide antisense to cPLA2 and methyl arachidonate fluorophosphate, a specific inhibitor of cPLA2. Biopsy and histochemical examination of erythematous sites expressed increased amounts of cPLA2 whereas nonerythematous irradiated sites did not. In contrast, cyclooxygenase-1 and -2 in cultures and skin explants were unaffected 6 h post-UV, and no change in cyclooxygenase activity was observed at this time. These results suggest that increased cPLA2 synthesis occurs only when skin is exposed to UV doses that are sufficient to cause erythema and indicate expression of cPLA2 participates in acute UV inflammation.

Amino Acid Sequence

Magnetic resonance imaging planes for the 3-dimensional characterisation of human coronary arteries.

We report a magnetic resonance imaging study which developed a consistent hierarchy of imaging planes for examination of the origins, courses and principal branches of the main coronary arteries of prepared human cadaveric hearts. The reference longitudinal axis was chosen between the aortic valve and the apex of the left ventricle. A series of transverse planes then successfully visualised the ostia of the left and right coronary arteries; the left main coronary, its bifurcation, and the left anterior descending artery for a distance 24 mm distal to its origin were clearly distinct in successively posterior sections as was the emergence and course of the right coronary artery. Further sections were derived from an axis that joined the posterior aspects of the left and right coronary artery ostia seen in cross-section, which demonstrated the origins of these arteries. They also traced the circumflex artery 30 mm beyond its point of emergence and demonstrated the course of the right coronary artery between the right ventricle and right atrium. The anatomical identifications were confirmed in selective 3-dimensional reconstructions of the cardiac anatomy around the aortic root and pulmonary artery origin. The orthogonal anatomical arrangements of the left and right coronary artery arterial trees thus permit a consistent set of imaging planes useful for the visualisation of all the major branches in a static heart in vitro. This may offer an approach useful for clinical imaging of human coronary vessels in vivo in the moving heart.

Aged

Electrogenic 2 Na/1 H antiport in crustacean epithelium is inhibited by a monoclonal antibody.

We have previously published evidence that suggests that Na/H exchange in crustacean and echinoderm epithelia occurs by an electrogenic antiporter protein with two external cation binding sites that accommodate Na, amiloride, or Ca and display a 2:1 monovalent cation antiport stoichiometry. The present study is an initial investigation into the molecular biology of this invertebrate electrogenic exchanger to ascertain its structural similarity to the analogous vertebrate electroneutral antiport system. A panel of monoclonal antibodies was prepared against components of lobster hepatopancreatic epithelial brush-border membranes and assayed immunohistochemically and by Western blotting. The antibodies were tested further in functional assays for their ability to interfere with electrogenic 2 Na/1 H antiport in isolated hepatopancreatic brush-border membrane vesicles. One cell line was identified producing an antibody that significantly inhibited the electrogenic exchange of cations by these membrane preparations and recognized a single protein band on Western blots of hepatopancreas, antennal gland, and gill epithelia corresponding to a molecular mass of 185 kDa. The existence of such an antibody probe may facilitate the purification of the electrogenic antiporter under denaturing conditions, in in vitro expression systems, or in prokaryotic expression libraries.

Animals