Bohdan Turyna

Biocompatibility of glass-crystalline materials obtained by the sol-gel method: Effect on macrophage function

The aim of this work was to confirm in vitro biocompatibility of a new gel-derived glass-crystalline material containing hydroxyapatite and wollastonite phases. For the purpose of comparison, studies were also carried out for a material of the same chemical composition obtained by the traditional melting method. We examined the behaviour and response of cells cultured in the presence of the studied materials. The level of activation of macrophages in culture was determined using three different methods: measurement of respiratory burst by chemiluminescence, nitrite assay and by bioassay of secreted cytokines after immunoelectrophoresis of acute phase proteins from hepatoma cells. All our results show a relatively low, close to control level, activation of macrophages exposed to the studied materials. This indicates a good biocompatibility of both the gel-derived material and the material obtained by the traditional melting method.

The ability of HDL to inhibit VCAM-1 expression and oxidized LDL uptake is impaired in renal patients.

OBJECTIVES This study examines the ability of HDL from hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) patients to suppress the expression of adhesion molecules in endothelial cells (ICAM-1, VCAM-1) and in monocytes (LFA-1, VLA-4) and to inhibit the uptake of oxidized LDL by macrophages. DESIGN AND METHODS Gene expression and the uptake of oxidized LDL were determined in 12 HD patients, 12 CAPD patients and 14 healthy volunteers. RESULTS HDL from renal patients were less effective than control lipoproteins in reducing VCAM-1 expression. HDL from CAPD patients inhibited LFA-1 expression to the highest extent. The ability of HDL from renal patients to reduce oxidized LDL uptake was lower compared to control group. CONCLUSIONS Decreased ability of HDL to suppress expression of VCAM-1 in endothelial cells and the uptake of oxidized LDL by macrophages can be one of the risk factors for atherosclerosis development in patients with renal failure.

Comparison of biocompatibility of gelderived bioactive ceramics in macrophage culture conditions

The behaviour of rat alveolar macrophages cultured in the presence of three new gel-derived ceramic biomaterials (CaO-P2O5-SiO2 system) with slightly different chemical compositions was examined. The abilities of these three materials to stimulate alveolar macrophages were compared. Non-treated and lipopolysaccharide-treated macrophages were used as control. The level of macrophage activation was determined by nitrite and prostaglandin E2 assay and respiratory burst measurement by chemiluminescence. The results of these studies showed different macrophage responses to these three relatively similar stimuli. Two of the studied materials were shown to be potent activators of respiratory burst and prostaglandin E2 secretion without any significant release of nitric oxide. On the contrary, the material characterized as the most surface reactive strongly affected only nitric oxide generation by the cells.

PREPARATION AND ELECTRON PARAMAGNETIC RESONANCE CHARACTERIZATION OF SPIN LABELED MONODERIVATIVES OF HORSE CYTOCHROME C

Horse cytochrome c was reacted with the spin label (succinimidyl-2,2, 5,5-tetra-methyl-3-pyrroline-1-oxyl-carboxylate) using optimized conditions and the reaction products were separated by a combination of cation-exchange chromatography and HPLC. The purified cytochrome c derivatives were digested with TPCK treated trypsin and the resulting peptides were separated by reverse phase HPLC. The modified Lys residues were subsequently characterized by Edman degradation and mass spectrometry. These analyses showed that five distinct cytochrome c derivatives had been produced which were modified at the specific Lys residues including Lys8, Lys25, Lys72, Lys86 or Lys87, respectively. The electron paramagnetic resonance (EPR) spectra for each cytochrome c derivative revealed that for the spin label attached to Lys8 and Lys87 only one component contributes to the spectrum whereas for Lys25, Lys72 and Lys86 the spectrum consists of two components. The highest mobility with the rotational correlation time, tauB, of 0.38 ns was observed for Lys87. The longest tauB of 1.84 ns was obtained for Lys72. An attempt to correlate the spin label mobility with the local protein structure is presented. These mono derivatized cytochrome c molecules provide a unique tool for EPR studying the interaction between cytochrome c and the lipid bilayer, as well as cytochrome c oxidase and reductase.

EPR studies of iso-1-cytochrome c: effect of temperature on two-component spectra of spin label attached to cysteine at positions 102 and 47

Abstract. Wild-type iso-1-cytochrome c from Saccharomycescerevisiae containing naturally occurring cysteine at position 102 and mutated protein S47C (derived from the protein in which C102 had been replaced by threonine) were labeled with cysteine-specific methanethiosulfonate spin label. Continuous wave (CW) electron paramagnetic resonance (EPR) was used to examine the effect of temperature on the behavior of the spin label in the oxidized and reduced forms of wild-type cytochrome c and in the oxidized form of the mutated protein. The computer simulations revealed that the CW EPR spectrum for each form of cytochrome c consists of at least two components [a fast (F) and a slow (S) component], which differ in the values of the rotational correlation times

Iron inhibits respiratory burst of peritoneal phagocytes in vitro.

\tau _{{\rm R}_\parallel }

LDL susceptibility to oxidation and HDL antioxidant capacity in patients with renal failure

(longitudinal rotational correlation time) and