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G Hofer

Publications and source records attributed to G Hofer.

13 recordsLinked to original sources

A new fluorescence method for the continuous determination of surface lipid oxidation in lipoproteins and plasma.

We report on a new method for the determination of lipid oxidation in lipoproteins and plasma. The biological lipid system is preloaded with a fluorescent analog of phosphatidylcholine containing diphenylhexatriene (DPH) propionic acid covalently linked to the sn-2 position. When externally added, the respective phospholipid label (DPHPC) localizes to the surface monolayer of a lipoprotein. Under oxidative conditions (e.g. in the presence of Cu2+ ions) the fluorophore undergoes decomposition, resulting in a continuous decrease of fluorescence intensity which reflects the oxidation of a chemically defined phospholipid molecule with well defined localization. When incorporated into LDL particles, the kinetics of the decrease in DPHPC fluorescence intensity upon exposure to Cu2+ us very similar to that of conjugated diene accumulation. Furthermore, our assay can be applied to follow the oxidation of lipids in diluted serum and may also be developed into a suitable test system for clinical studies of susceptibility of plasma lipids to oxidation.

Diphenylhexatriene

Specific and non specific stimulation of prostaglandin release by human skin fibroblasts in culture.--Are changes of membrane fluidity involved?

In order to study a bidirectional relationship between changes of membrane fluidity and prostaglandin synthesis, the arachidonic acid cascade was stimulated in cultured human skin fibroblasts by unspecific stimuli (hypotonicity, low calcium concentrations) and by the specific stimulus, bradykinin. Fluorescence anisotropy of trimethylammoniumdiphenylhexatriene was used to measure membrane fluidity in cell monolayers. Hypotonicity or low calcium concentrations induce membrane fluidisation and prostaglandin synthesis. However, after specific stimulation of prostaglandins with bradykinin (at normocalcic and isotonic conditions) a rigidification of plasma membranes was observed in living cells. Fluidisation of membranes and bradykinin activate phospholipase A2 and induce prostaglandin synthesis. Although in cell membrane preparations increased phospholipase A2 activity leads to fluidisation, in our model a membrane fluidisation was not observed after stimulation of phospholipase with bradykinin. This suggests that in living cells a fluidizing effect of lysolecithin resulting from phospholipase A2 activation may be rapidly counteracted by its removal. A decrease of phosphatidylcholin content and consequently a rigidification of the membrane may ensue. Thus, the cell culture model using two different ways of stimulating phospholipase activity, helps to define the directional relationship between changes of membrane fluidity and activation of phospholipase and the arachidonic acid cascade in living human cells.

Bradykinin

Radioimmunoassay for immunoreactive non-collagenous domain of type IV collagen (NC1) in serum: normal pregnancy and preeclampsia.

The use of a magnetic separation agent in a modified radioimmunoassay for the non-collagenous cross-linked region (NC1) of basement membrane collagen is described. The rabbit anti-NC1 serum employed in the assay revealed a binding behaviour similar to that reported recently (Mark et al. (1985) Eur. J. Biochem. 146, 555-562; Schuppan et al. (1986) J. Clin. Invest. 78, 241-248) with respect to the integrity of NC1-disulphide bridges, affinity to NC1 subunits and lack of reactivity with other determinants of type IV collagen. Immunoreactive serum NC1, which increased towards the end of gestation, showed a broad peak during the second trimester of pregnancy. In preeclampsia, immunoreactive serum-NC1 was slightly elevated.

Blotting, Western