Biomedical subjects

J VanSteveninck

Publications and source records attributed to J VanSteveninck.

31 records · Page 2Linked to original sources

PubMed1996

Synergistic interaction of photodynamic treatment with the sensitizer aluminum phthalocyanine and hyperthermia on loss of clonogenicity of CHO cells.

When CHO cells were exposed to hyperthermia and subsequently to photodynamic treatment, the combined effects were additive but in the reverse sequence the interaction was synergistic. The synergistic interaction comprised two quite different components: (1) photodynamically induced sensitization of cellular proteins and/or supramolecular structures for thermal inactivation and (2) a photodynamically induced inhibition of the cellular repair system for sublethal thermal damage. The first component of the synergistic interaction was reflected by a change of the Arrhenius parameters of thermal cell killing. A lowering of the activation energy of this process was responsible for the synergistic interactions, whereas a concomitant decrease of the frequency factor, opposing this effect, actually caused a much lower degree of synergism at higher temperatures. This component of the synergistic interaction did not respond to the insertion of an intermediate incubation period between the two treatments. The second component of the synergistic interaction, viz the interference with the ability of cells to survive sublethal thermal damage, was reversible, as an intermediate incubation between photodynamic treatment and hyperthermia resulted in its repair. The photodynamically induced inhibition of the ability of cells to survive sublethal thermal damage was not related to ATP or glutathione depletion, inhibition of de novo protein synthesis or impairment of degradation of damaged protein molecules. Restoration of the repair system for sublethal damage depended on a metabolic process and required free intracellular Ca2+, suggesting that a cell signaling pathway may be involved. Thus, in a practical sense the magnitude of the synergistic interaction between photodynamic treatment and hyperthermia depends on the length of the interval between the two treatments and on the temperature and duration of the subsequent thermal treatment. This may have significant consequences for the development of clinical protocols for the combined application of photodynamic therapy and hyperthermia in the treatment of tumors.

Animals↗

PubMed1995

Factors affecting the amount and the mode of merocyanine 540 binding to the membrane of human erythrocytes. A comparison with the binding to leukemia cells.

In the presence of albumin Merocyanine 540 (MC540) exhibits a very limited binding to the outer surface of the membrane of normal erythrocytes, whereas pronounced binding is observed to leukemia cells. To find out whether this difference is due to differences in the composition or structural organization of the cell membrane we analyzed effects of a number of covalent and non-covalent perturbations of the red cell membrane on the binding and fluorescence characteristics of membrane-bound MC540. It is shown that exposure of the cells to cationic chlorpromazine, neuraminidase or photodynamic treatment with AlPcS4 as sensitizer caused a limited increase (30-50%) of MC540 binding, together with a red shift of the fluorescence emission maximum and an increase of the relative fluorescence quantum yield of membrane-bound MC540. Other forms of perturbation of the membrane structure, like hyperthermia (48 degrees C) and treatments that produce a decrease of phospholipid asymmetry in addition to accelerated flip-flop, did not result in increased MC540 binding, but did cause a red shift of the fluorescence emission maximum and an increase of the relative fluorescence quantum yield. These changes in fluorescence properties indicate a penetration of the dye into more hydrophobic regions in the membrane. MC540, bound to Brown Norway myelocytic leukemia cells, exhibited a red shift of the fluorescence emission maximum and an increased relative fluorescence quantum yield as compared to MC540 bound to untreated erythrocytes. These changes were of the same order of magnitude as in photodynamically treated red blood cells. Dye binding per surface area, however, was about 3-times higher with these leukemia cells than with photodynamically treated red blood cells. This demonstrates that certain perturbations of the erythrocyte membrane evoked a MC540 binding that became qualitatively comparable to the dye binding to leukemia cells, although dye binding per surface area was still significantly lower.

Cell Membrane↗

PubMed1995

Interaction of photodynamically induced cell killing and dark cytotoxicity of rhodamine 123.

Loss of clonogenicity of Chinese hamster ovary (CHO) cells, murine L929 fibroblasts and human bladder carcinoma T24 cells caused by photodynamic treatment (PDT) with hematoporphyrin derivative (HPD) is synergistically enhanced by subsequent incubation with rhodamine 123 in the dark. For CHO and L929 cells this synergistic interaction can be explained by an increased uptake of rhodamine 123 as the result of the photodynamic treatment. With aluminum phthalocyanine (AIPC) as photosensitizer only additive effects were observed in the three cell lines. Incubation in the dark with rhodamine 123, followed by a photodynamic treatment with HPD, resulted in an antagonistic interaction with regard to loss of colony formation. With AIPc the combination of treatments resulted in an additive effect with L929 and T24 cells, whereas with CHO cells a slight antagonistic interaction was observed. An antagonistic effect was also observed in model experiments, treating histidine photodynamically with HPD and measuring oxygen consumption. A possible explanation of these results could be an interaction or complex formation of rhodamine 123 with HPD resulting in a diminished singlet oxygen production. With AIPc this does not take place.

Animals↗

PubMed1994

The role of DNA damage and inhibition of poly(ADP-ribosyl)ation in loss of clonogenicity of murine L929 fibroblasts, caused by photodynamically induced oxidative stress.

Reactive oxygen species are used to eradicate malignant cells in photodynamic therapy as well as in other cancer therapies. Despite many efforts, the pathways leading to cellular damage and cell killing due to the action of these species are poorly understood. In previous studies with hematoporphyrin derivative-sensitized L929 murine fibroblasts, the only parameter for which a relation with photodynamically induced reproductive cell death could not be excluded was inhibition of DNA excision repair. The present results show that loss of clonogenicity of these cells in fact is related to a series of effects, including the development of slight, irreperable DNA damage, a virtually complete inhibition of poly(ADP-ribosyl)ation activation, a transient elevation of the intracellular calcium concentration and, after a lag time of about 8 h, DNA fragmentation caused by endonuclease activity. This conclusion is supported by the observation that photodynamic treatment inhibited the repair of X-ray-induced DNA strand breaks and suppressed X-ray- and methyl methanesulfonate-induced enhancement of poly(ADP-ribosyl)ation. Our experimental results further suggest that in this cell line the photodynamically induced inhibition of enhanced poly(ADP-ribosyl)ation could well be involved in inhibition of repair of DNA strand breaks and in activation of endonuclease activity.

Animals↗

PubMed1994

Effects of the micro-environment on the photophysical properties of hematoporphyrin.

The photochemical degradation of histidine, cysteine and tyrosine with hematoporphyrin as sensitizer was potentiated by the presence of Sephadex, BioGel or Percoll particles. This effect could only partly be explained by binding of the sensitizer to the gel particles, leading a.o. to monomerization of aggregated sensitizer molecules in the aqueous environment. The hematoporphyrin triplet state life time increased from 250 microseconds in phosphate buffer to 1992 microseconds in the presence of 50% Percoll. Most likely the effect of the gel particles on the sensitizer triplet state is, at least partly, mediated by the solvent. A plausible explanation seems to be that the vicinal water structure at the particle interface stabilizes the sensitizer triplet.

Cysteine↗

PubMed1994

Characterization of exocytosis in electropermeabilized neutrophils by flow cytometric analysis: difference in sensitivity to calcium and guanosine-5'-[gamma-thio]triphosphate.

When rabbit neutrophils were subjected to two electrical discharges of 4.75 kV/cm, the cells became permeable to propidium iodide. Measurement of propidium iodide fluorescence using flow cytometry showed that all cells in the suspension were permeabilized. The cells remained permeable for > 20 min when the cells were stored at 0 degree C. When exocytosis was induced by Ca2+ alone, the orthogonal light scatter (a sensitive parameter for cell granularity) of the complete population changed depending on the concentration. All the cells were equally sensitive to Ca2+ and showed a similar degree of exocytosis at the same time. In the presence of a fixed concentration of Ca2+ and a variable concentration of guanosine-5'-[v-thio]triphosphate (GTP gamma S), a division of the cell population was observed in the orthogonal light scatter histogram. At low GTP gamma S concentrations, a part of the population showed complete exocytosis and a part of the population showed almost no exocytosis. With increasing GTP gamma S concentrations, the light scatter pattern of the population changed indicating that the cells were gradually sensitive to GTP gamma S. Electropermeabilized neutrophils showed an equal sensitivity to Ca2+ and a graded sensitivity to GTP gamma S. Flow cytometry is considered as an ideal tool to study such an effect on a cell-to-cell basis.

Animals↗

PubMed1994

Hematoporphyrin derivative-induced photodynamic inhibition of Na+/K(+)-ATPase in L929 fibroblasts, Chinese hamster ovary cells and T24 human bladder transitional carcinoma cells.

The photodynamically induced inhibition of Na+/K(+)-ATPase with hematoporphyrin derivative as photosensitizer was studied in murine L929 fibroblasts, Chinese hamster ovary (CHO) cells and T24 human bladder transitional carcinoma cells. In T24 cells the inhibition of the enzyme activity appeared to be caused by ATP depletion rather than by direct damage from the enzyme itself. In L929 and CHO cells, on the other hand, the inhibition was caused by photodynamic damage from the enzyme molecule. For all three cell lines it was shown that a causal relationship between photodynamically induced reduction in Na+/K(+)-ATPase activity and the loss of clonogenicity is highly unlikely.

Animals↗

PubMed1993

HPD-induced photodynamic changes in intracellular cyclic AMP levels in human bladder transitional carcinoma cells, clone T24.

Human bladder transitional carcinoma cells, clone T24, were incubated with hematoporphyrin derivative and subsequently exposed to red light. Immediately after light exposure a light-dose dependent increase in intracellular cyclic AMP levels was observed. The concentrations decreased to non-treated levels within 2 h after illumination. Inhibition of cyclooxygenase with indomethacin reduced the increase drastically, whereas the presence of PGE2 enhanced the production of cyclic AMP. PGE2, 8'-bromo-cyclic AMP and a direct stimulation of adenylyl cyclase by forskolin reduced the photodynamically induced cell killing. On the other hand, suppression of cyclic AMP by the cyclooxygenase inhibitor indomethacin, enhanced the photodynamically induced cell killing. Therefore, it is suggested that cyclic AMP production in T24 cells is a step in a cellular rescue pathway, resulting in protection against photodynamic cell killing.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1993

Neutrophil chemotaxis induced by the diacylglycerol kinase inhibitor R59022.

The diacylglycerol kinase inhibitor R59022 induced chemotaxis in neutrophils. The response to R59022 was primarily chemotactic and only very little chemokinetic. Pretreatment with the protein kinase C inhibitors staurosporine and AMG-C16 inhibited chemotaxis induced by R59022 indicating the involvement of protein kinase C. In contrast, chemotaxis induced by fMet-Leu-Phe was only slightly inhibited by staurosporine and AMG16. The effects of R59022 were comparable to the effects of the protein kinase C activators DiC8 and PMA and suggest an involvement of protein kinase C. Pretreatment with pertussis toxin inhibited R59022-induced migration, fMet-Leu-Phe-induced migration, and random migration. GTP gamma S, which stimulates migration of electropermeabilized neutrophils by itself, causes an additive increase of migration in electropermeabilized neutrophils stimulated with a suboptimal concentration R59022, but causes a synergistic increase of migration in cells stimulated with a suboptimal concentration fMet-Leu-Phe. The effects of GTP gamma S on migration are completely inhibited by AMG-C16. This suggests that the GTP-binding protein involved in R59022-activated migration is the G protein that is associated with random migration.

Animals↗

PubMed1993

Ca(2+)-mediated prostaglandin E2 induction reduces haematoporphyrin-derivative-induced cytotoxicity of T24 human bladder transitional carcinoma cells in vitro.

The effects of haematoporphyrin-derivative-mediated photodynamic treatment on arachidonic acid metabolism and its relation to clonogenicity have been studied in human bladder-tumour cells. Photodynamic treatment resulted in a transient release of arachidonic acid-derived compounds; prostaglandin E2 (PGE2) and thromboxane B2 (TXB2) especially were strongly increased. This release was reduced by chelation of intracellular Ca2+ with Quin-2 or by lowering the extracellular Ca2+ concentration in the medium with EGTA, presumably resulting in inhibition of phospholipase A2. A similar reduction was obtained when indomethacin, an inhibitor of the cyclo-oxygenase pathway, was added prior to light exposure. These three treatments enhanced the photosensitivity, as revealed by the clonogenicity assay. Incubation with PGE2 prior to light exposure, but not with TXB2, protected against reproductive-cell death. The results of these experiments suggest that Ca(2+)-mediated activation of cyclo-oxygenase, resulting in increased levels of PGE2, participates in a cellular-defence mechanism against photodynamic cell killing.

Calcium↗

PubMed1993

Strontium and barium induce exocytosis in electropermeabilized neutrophils.

Calcium, strontium and barium induced an exocytotic response in electropermeabilized rabbit neutrophils while magnesium was without any effect. The extent of enzyme release was found to depend upon the concentration of these cations. For all cations, an optimum concentration was found with the same maximum enzyme release. At concentrations higher than optimum a decrease in lysozyme release was observed. Efficiency to induce enzyme release was in the order: Ca2+ > Sr2+ > Ba2+. Enzyme release was significantly enhanced by guanosine-5'-[gamma-thio]triphosphate (GTP gamma S) resulting in a shift to the left of the dose/response curve. The enhancement by GTP gamma S was strongest with Ca2+, was less with Sr2+, and was very little with Ba2+. The time course of lysozyme release was the same for Ca2+, Sr2+, and Ba2+ in the presence and absence of GTP gamma S when suboptimal cation concentrations were used. A decrease in responsiveness to the effectors after electropermeabilization was observed with Ca2+, Sr2+ and Ba2+ in the presence and absence of GTP gamma S. The lysozyme release induced by the different cations was not inhibited by the protein kinase C inhibitor staurosporine and was slightly affected by pertussis toxin. Ca2+ and Sr2+, but not Ba2+, potentiated formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe) induced enzyme release in intact neutrophils. The divalent cation ionophore A23187 induced enzyme release in the presence of Ca2+ and Sr2+ but not in the presence of Ba2+. The results obtained with electropermeabilized neutrophils indicate that Sr2+ and Ba2+ can act as substitutes for Ca2+ in activating exocytosis, and that permeabilized neutrophils provide the best tool to investigate the effects of alkaline earth ions in exocytosis.

Animals↗

PubMed1993

GTP[S] stimulates migration of electropermeabilized neutrophils via a pertussis toxin-sensitive G-protein.

Electropermeabilized neutrophils were used to study the role of G-proteins in neutrophil migration. Rabbit neutrophils, under specific conditions, retained their ability to migrate after electropermeabilization. Introduction of guanosine-5'-[3-thio] triphosphate (GTP[S]) into the cell interior stimulated random migration and enhanced migration activated by a suboptimal concentration of formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe) (10(-11) M). GTP[S] had no effect on random migration by intact cells, or on migration of intact cells activated with a suboptimal concentration of fMet-Leu-Phe, indicating that the effect of GTP[S] was intracellular. The effects of GTP[S] were inhibited by pertussis toxin and by guanosine-5'-[2-thio] diphosphate (GDP beta S) indicating that a pertussis toxin-sensitive G-protein was involved. GTP stimulated random migration to the same extent as GTP[S], but had only a small effect on migration activated by a suboptimal concentration of fMet-Leu-Phe (10(-11) M). Several other nucleotides tested had no effect on random migration or migration activated with 10(-11) M fMet-Leu-Phe. The results show that neutrophil migration can be potentiated by direct activation of a pertussis toxin-sensitive G-protein, and the results obtained with GTP suggest that possibly more than one G-protein is involved in this process.

Animals↗

PubMed1992

Interaction of photodynamic treatment and either hyperthermia or ionizing radiation and of ionizing radiation and hyperthermia with respect to cell killing of L929 fibroblasts, Chinese hamster ovary cells, and T24 human bladder carcinoma cells.

Both hyperthermia and photodynamic therapy of cancer are frequently used in combination with other treatment modalities in order to improve tumor control with minimal damage to normal tissues. The present results indicate that the most effective combination of treatment modalities is different in different cell types. For instance, ionizing irradiation and hyperthermia exhibited additivity when applied to L929 fibroblasts, in contrast to the synergistic interaction described before in many other cell lines. This aberrant behavior of L929 cells could be explained by the relative insensitivity of DNA repair in these cells to hyperthermia. Conversely, a synergistic interaction between photodynamic treatment and ionizing irradiation was observed with L929 fibroblasts, whereas these treatments were additive with Chinese hamster ovary and T24 cells. The synergistic interaction with L929 cells could be explained by the high sensitivity of DNA repair in these cells to photodynamic treatment. Photodynamic treatment and hyperthermia exhibited a synergistic interaction in L929, Chinese hamster ovary, and T24 cells. The critical target for cell killing by the combined treatment protocol in these cell lines has not yet been elucidated. In all three cell lines, however, analysis of the results according to the Arrhenius equation revealed a photodynamically induced change of both the frequency factor and the activation energy of subsequent thermal cell killing. It is considered that this may indicate a basic mechanism, in which a particular protein is a common, critical target of the two modalities of treatment.

Animals↗