Radiation damage on Langmuir monolayers of the anionic 1.2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (sodium salt)(DPPG) phospholipid at the air-DNA solution interface.

The resilience of cells to ultraviolet (UV) irradiation is probably associated with the effects induced in biological molecules such as DNA and in the cell membrane. In this study, we investigated UV damage to the anionic 1.2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (sodium salt) (DPPG) phospholipid, which is an important component of cell membranes. In films cast from DPPG emulsions, UV irradiation induced cleavage of C-O, C=O and -PO(2-) bonds, while in Langmuir monolayers at the air/water interface representing the cell membrane this irradiation caused the monolayer stability to decrease. When DNA was present in the subphase, however, the effects from UV irradiation were smaller, since the ionic products from degradation of either DPPG or DNA stabilize the intact DPPG molecules. This mechanism may explain why UV irradiation does not cause immediate cell collapse, thus providing time for the cellular machinery to repair elements damaged by UV.

Radiation-induced free-radical transformation of phospholipids: MALDI-TOF MS study.

Under the action of free-radical reaction initiators on membrane phospholipids, complex processes are taking place in both hydrophobic and hydrophilic parts of the phospholipids. Realization of these processes results in a mixture consisting of the initial lipids and their peroxidation and fragmentation products. Identification of compounds in such mixtures requires analytical methods of high sensitivity, reproducibility and accuracy to be applied. These properties are characteristic of the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) method. In the studies of radiation-induced free-radical transformations of phosphatidylglycerol, the MALDI-TOF MS in combination with thin layer chromatography (TLC) has been shown to be able to detect and identify products of free-radical transformations taking place in both hydrophilic and hydrophobic parts of the phospholipid. Thus, the MALDI-TOF MS can serve as a suitable analytical tool to investigate free-radical transformations of lipids.

Gamma-irradiation of liposomes composed of saturated phospholipids: effect of bilayer composition, size, concentration and absorbed dose on chemical degradation and physical destabilization of liposomes.

Liposomes composed of dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylglycerol (DPPG), or mixtures of these two phospholipids were exposed to gamma-irradiation in an air environment. Disappearance of the mother compounds was monitored by HPLC analysis. Plotting of the logarithmic values of residual DPPC or DPPG concentration versus irradiation dose resulted in straight lines. The slopes of these lines (overall degradation constants) depended on the type of phospholipids, concentration of the liposomes and the size of the liposomes. Under the chosen conditions, addition of DPPG in DPPC-liposomes did not affect the degradation rate constant of DPPC and vice versa. The presence of phosphate buffer (pH 7.4), pH or presence of sodium chloride did not affect the irradiation damage either. Minor changes were found upon analysis of total fatty acids by GLC and upon measurement of water soluble phosphate compounds. These changes were less pronounced than the changes monitored by HPLC of phospholipids, because the HPLC analysis monitored the overall degradation of the liposomal phospholipids. Thin-layer chromatography/fast atom bombardment mass spectrometry (TLC/FAB-MS) analysis of irradiated and non-irradiated DPPC or DPPG provided information on the structure of several degradation products. Degradation routes which include these degradation products are proposed. Gamma-irradiation neither affected the size of the liposomes nor the bilayer rigidity as determined by dynamic light scattering and fluorescence anisotropy of the probe 1,6-diphenyl-1,3,5-hexatriene (DPH), respectively. However, upon gamma-irradiation, changes in the melting characteristics of the liposomes were found by differential scanning calorimetry (DSC) measurements. The pre-transition melting enthalpy of the liposomal bilayer decreased or disappeared and the main-transition broadened. The changes found in DSC scans correlated qualitatively well with the changes recorded after HPLC analysis of phospholipids.

A comparison of X-irradiation and ferrous ion-ascorbate on oxidation of phosphatidylglycerols in multilamellar liposomes.

The effect of ferrous ion-ascorbate and X-radiation on multilamellar liposomes, composed of either completely saturated, unsaturated or a mixture of saturated and unsaturated fatty acids, is reported. Lipid composition is shown to be of critical importance in determining the extent to which peroxidation occurs. Liposomes composed of the mixture of saturated and unsaturated fatty acids are peroxidized to a lesser extent by ferrous ion-ascorbate. The reduced peroxidation is apparently the result of an inhibition mechanism shown by the saturated lipid component. In contrast, liposomes composed of mixed lipids do not reduce the level of peroxidation induced by ionizing radiation. These results show that the composition of liposomes plays a role in determining the extent to which peroxidation occurs when a chemical oxidant is employed, but composition is a negligible factor when ionizing radiation is the oxidant.