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M Raes

Publications and source records attributed to M Raes.

At least 55 records · Page 3Linked to original sources
PubMed1994

Induction of cyclooxygenase by interleukin 1: comparative study between human synovial cells and chondrocytes.

OBJECTIVE: To investigate the regulation of the prostaglandin (PG) synthesis by interleukin 1 (IL-1) in human synovial cells and chondrocytes. METHODS: Both cell types stimulated by human recombinant IL-1 synthesized PGE2, PGF2 alpha and 6-keto-PGF1 alpha. RESULTS: PGE2 was the major PG synthesized. When arachidonic acid was added exogenously at the end of the stimulation, an increase in the prostaglandin synthesis was observed after 6 and 24 h suggesting that cyclooxygenase is the limiting enzyme. Using actinomycin D and cycloheximide, PG synthesis was shown to be protein synthesis dependent. Inhibition of the constitutive cyclooxygenase by aspirin before the IL-1 stimulation confirmed that the increased prostaglandin synthesis was due to a de novo synthesis of cyclooxygenase. CONCLUSION: This enzyme induction by IL-1 was found to be similar in both cell types.

Arachidonic Acid↗
PubMed1993

Routine prostaglandin assay by GC-MS in multiwell tissue culture plates: application to human synoviocytes and chondrocytes.

Prostanoids can be assayed by the highly sensitive and reproducible gas chromatography-negative ion chemical ionization-mass spectrometry (GC-NICI-MS) technique. This paper describes a simplified method applied to the quantification of prostaglandins (PG) in the supernatants of limited amounts of cells, cultured in multiwell plates (30,000 cells/well). The culture medium was extracted on C2 columns and the derivatization was performed in three steps. Prostanoids were then analyzed by GC-NICI-MS within 10 min. The storage conditions at each step were tested in order to achieve maximal yields. This procedure allowed the determination and quantification of PG produced by cultured human synovial cells and chondrocytes at the basal level as well as after interleukin-1 stimulation. PGE2, PGF2 alpha, and 6-keto-PGF1 alpha, the stable form of PGI2, were the main PG detected. Derivatization and GC-MS analyses of 18 samples could easily be performed in 1 day. The simplified GC-NICI-MS method can thus be routinely applied for assaying PG in any cell line cultured in multiwells.

Calibration↗
PubMed1993

Differential effects of interleukin-1 alpha and beta on the arachidonic acid cascade in human synovial cells and chondrocytes in culture.

The effects of interleukin-1 alpha and beta were tested on the [3H]-arachidonic acid release and the prostaglandin synthesis by human cultured synovial cells and chondrocytes. Both forms of interleukin-1 stimulated the arachidonic acid release but interleukin-1 beta was more potent than IL-1 alpha. Human synovial cells and chondrocytes synthesized three types of prostaglandins upon stimulation with interleukin-1 alpha or beta: prostaglandin E2, F2 alpha and 6-keto-prostaglandin F1 alpha. Regarding the synthesis of these prostaglandins, IL-1 beta was again more potent than IL-1 alpha. A comparison between interleukin-1-stimulated synovial cells and chondrocytes revealed neither significant quantitative nor qualitative differences in both the arachidonic acid release and the prostaglandin synthesis.

Arachidonic Acid↗
PubMed1993

Lymphocyte subpopulations in healthy newborn infants: comparison of cord blood values with values five days after birth.

Significant differences (p < 0.0001) were demonstrated in lymphocyte subpopulations both in cord blood and in venous blood samples obtained at day 5 from the same healthy infants. Numbers of T lymphocytes increased, especially CD4+/CD45RA+ cells, whereas numbers of B lymphocytes and natural killer cells decreased without changes in CD8+ and other cytotoxic cells.

Female↗
PubMed1992

Importance of various antioxidant enzymes for cell stability. Confrontation between theoretical and experimental data.

A theoretical model was developed taking into account the production and destruction of oxygen-derived free radicals. The steady state of the system was derived by using the rate equations of these reactions, and the stability of the system was tested. In the simplified model, only one stable steady state was found. However, we know that glutathione peroxidase can be inhibited by hydroperoxides, and, when incorporated into the model, this effect led to a complex situation with the presence of some stable and some unstable domains according to the concentration of either the enzyme or the hydroperoxide. This qualitative description of the system was compared with experimental data on the protection given by three antioxidant enzymes, and concordance of data was found which allows some quantification of the system. A general view of the efficiency of the three antioxidant enzymes and of the stability of the system according to their concentrations could be produced.

Animals↗
PubMed1992

The importance of antioxidant enzymes in cellular aging and degeneration.

Aerobic cells contain various amounts of the three main antioxidant enzymes: superoxide dismutase (SOD), catalase and GSH peroxidase. These three enzymes are necessary for cell survival since inhibition of their activity leads to the arrest of cell mitosis and to cell death. Amongst them, GSH peroxidase was shown to be more efficient than catalase and much more than SOD. This result was obtained by comparing the cell protection against oxidative stress after their microinjection in the cytoplasm. With age, the level of these antioxidant enzymes does not change in several experimental models, so that it is not possible to explain the aging process by a lack of protection due to a decrease in the activity of these three enzymes. However, tissues and cells are more susceptible to free radical attacks with age. In order to understand the importance of free radicals in this process, we have to distinguish between their respective effects on cell mitosis, cell death and cell aging. The effects on mitosis and cell death are well described, and the results clearly show a threshold of response which is determined by the antioxidant content of the cell. There is now evidence that short free radical stresses can also speed up the aging of in vitro cultured human fibroblasts. However, such effects are not typical of free radicals but are also obtained with many other deleterious substances so that free radicals have to be considered as one amongst other factors responsible for influencing the evolution of a cell to an older stage or to cell death. The lowering of the general metabolism and of the free energy in old cells are probably the main factors responsible for the increased susceptibility of these cells to stresses such as oxidative stresses.

Aerobiosis↗
PubMed1991

Aging as a multi-step process characterized by a lowering of entropy production leading the cell to a sequence of defined stages.

The principles of the thermodynamics of irreversible processes which occur in biological cells considered as open systems, have been reviewed and applied in order to describe a possible evolution of cells during aging. The main feature of such an approach is that cells operating in a steady state, optimize their free energy production by lowering their entropy production which is kept to a minimum. Instabilities can however occur which can lead the cell from one steady state to another characterized by a lower production of entropy. Concomitantly, the level of errors or entropy of the system will increase. The process will continue and the cell will go from one state to another until a critical level is attained where the cell can not cope any more with keeping its organization and will die. The effect of stresses and mitosis have been also considered in this model. Such an approach stresses that cells can only subsist as a whole in certain states which are the result of the genetic constitution but also of the optimalization of cellular functions given their requirement in energy and the fluctuations from the environmental changes. In this respect, it reconciles both programmed and stochastic theories of aging.

Aging↗
PubMed1991

Association of antioxidant systems in the protection of human fibroblasts against oxygen derived free radicals.

The protection of human diploid fibroblasts against high oxygen tension was investigated using various combinations of the three major antioxidant enzymes: superoxide dismutase, catalase and glutathione peroxidase. alpha-Tocopherol, a well-known hydrophobic antioxidant, was also tested in combination with the different enzymes. Microinjection of solutions containing different combinations of the three enzymes was compared with the injection of each single enzyme. We observed that the protections given by catalase or superoxide dismutase on the one hand, and by glutathione peroxidase on the other hand, were additive. Surprisingly, the combinations of catalase and superoxide dismutase were less effective than catalase alone and was even toxic at low SOD concentrations. Addition of alpha-tocopherol following the injection of any of the three enzymes was highly beneficial, but the strongest synergistic effect was obtained with glutathione peroxidase. These results stress the importance of membrane protection by alpha-tocopherol and indirectly by glutathione peroxidase. They also showed that any injection leading to the decrease in the O2.- or H2O2 concentration combined with one of these two protectors is very beneficial for the cells probably by decreasing the OH concentration. This is also proven by the very good protective effect obtained with desferrioxamine.

Antioxidants↗
PubMed1990

Respiratory activity of isolated rat liver mitochondria following in vitro exposure to oxygen species: a threshold study.

Respiratory activity of isolated rat liver mitochondria was assayed following in vitro exposure to oxygen radicals. Our results show that mitochondrial respiration is more sensitive to O2.(-) than to H2O2. However, ferrous ions drastically enhance the toxicity of the enzymatic system generating H2O2 because of the production of the hydroxyl radicals. A protection against those oxygen species could be given by SOD in the xanthine/xanthine oxidase system and by catalase with the glucose/glucose oxidase system. The most damaging system was the combination of Fe2+ with H2O2. In this case, OH. is formed in a Fenton-like reaction. The fact that the OH. is the most damaging molecule accounts for the finding that catalase and desferrioxamine were efficient protectors in this system. Threshold levels of O2.(-) and H2O2 able to inhibit the mitochondrial respiration have been estimated. It is concluded that under normal respiration such thresholds are not reached in vivo and that the impairment of the mitochondrial respiratory activity does not seem to originate only from the natural free radical production in those organelles. However, if the production of free radicals is such to exceed the defense capability, like under oxidative stress, then the critical threshold can be surpassed and the respiration impaired leading to irreversible damages.

Aging↗
PubMed1990

Glutathione peroxidase, superoxide dismutase, and catalase inactivation by peroxides and oxygen derived free radicals.

Glutathione peroxidase (GPX), superoxide dismutase (SOD) and catalase are the most important enzymes of the cell antioxidant defense system. However, these molecules are themselves susceptible to oxidation. The aim of this work was to estimate to what extent this system could be inactivated by its own substrates. We tested the effect of hydrogen peroxide, cumene hydroperoxide, t-butyl hydroperoxide and hydroxyl and superoxide radicals on GPX, SOD and catalase. For GPX, a 50% inactivation was observed at 10(-1) M (30 min, 37 degrees C) for hydrogen peroxide, 3 x 10(-4) M (15 min, 37 degrees C) for cumene hydroperoxide and 5 x 10(-5) M (11 min, 37 degrees C) for t-butyl hydroperoxide. Unlike the hydroxyl radicals, superoxide anions did not inactivate this enzyme. Catalase was inactivated by hydroxyl radicals and by superoxide anions but organic peroxides had no effect. SOD was inactivated by 50% by hydrogen peroxide at 4 x 10(-4) M (20 min, 37 degrees C), but organic peroxides and hydroxyl radicals were ineffective on this enzyme. Since the three enzymes of the antioxidant system are susceptible to at least one of the oxidative reactive molecules, in the case of high oxidative stresses such an inhibition could take place, leading to an irreversible autocatalytical process in which the production rate of the oxidants will continuously increase, leading to cell death.

Catalase↗
PubMed1990

Importance of a threshold for error accumulation in cell degenerative processes. I. Modulation of the threshold in a model of free radical-induced cell degeneration.

Antioxidant enzymes (catalase, superoxide dismutase and glutathione peroxidase) have been injected into human fibroblasts exposed to 2 atm O2 in order to test if the threshold of oxidative damage versus antioxidant defenses could be modulated and if the damage remains reversible beyond the threshold. Cell damage was estimated by thymidine incorporation and cell survival curves. The proportion of dividing cells, measured by thymidine incorporation, rapidly decreased after O2 incubation: no cells could divide after 15 h of hyperoxia. However, cells incubated for a short time and injected with a high concentration of any of the three enzymes divided like non-oxygen-incubated cells: the enzymes could protect the cells against their loss of division potential. However, when cells were incubated for a longer period and/or when the injected enzyme concentration was lower, cells were either less or not protected and could no longer divide. These results suggest the presence of a threshold for the oxidative damage which cannot be totally repaired and which impairs the cell division; this threshold can, however, be modulated by supplementation of antioxidant enzymes, glutathione peroxidase being the most efficient.

Catalase↗
PubMed1989

Long-term effect of ipratropium bromide and fenoterol on the bronchial hyperresponsiveness to histamine in children with asthma.

We studied the effects of the anticholinergic ipratropium bromide (40 micrograms three times daily) and the beta-agonist, fenoterol (0.2 mg three times daily), both administered by powder inhaler, on bronchial hyperresponsiveness (BHR) to histamine in children, aged 7 to 15 years with mild stable asthma and limited bronchoconstriction who had a highly increased BHR. The double-blind, randomized, parallel study was conducted and performed in spring and early summer. BHR and FEV1 were measured on two occasions, before the start of treatment and monthly thereafter for 4 months. Symptoms, peak expiratory flow, and concomitant medication were registered daily. Nine of the 12 patients receiving ipratropium bromide and all eight patients receiving fenoterol completed the study. Patients completing treatment had few symptoms and were in a stable condition throughout the treatment period. Neither the administration of ipratropium bromide nor fenoterol resulted in a significant change of BHR. We concluded that long-term treatment with ipratropium bromide or fenoterol had no effect on BHR in children with mild stable asthma.

Adolescent↗
PubMed1988

Microinjection of antibodies against superoxide dismutase and glutathione peroxidase.

Antibodies were prepared against glutathione peroxidase, superoxide dismutase, and catalase. Inhibition of the enzyme activity was obtained with anti-Gpx and anti-SOD antibodies but not with anti-CAT antibodies. The antibodies were then injected into human fibroblasts and bovine chondrocytes in culture either under normal conditions or under 1 atm of oxygen. The injected anti-Gpx and anti-SOD antibodies increased the mortality rate of the fibroblasts incubated under 1 atm of oxygen. However, when cells were incubated under normal atmosphere, anti-Gpx antibodies inhibited the division while anti-SOD antibodies increased this capacity. Anti-Gpx antibodies injected into chondrocytes decreased their viability. Injection of control antiserum had no effect. These data stress the primary importance of Gpx as antioxidant under all conditions and the relative efficiency of SOD according to the balance between the radical production and the activity of the other antioxidant systems.

Animals↗
PubMed1988

Effect of procaine on cultivated human WI-38 fibroblasts.

Procaine is a local anesthetic, also used in experimental gerontology and has been tested in cultivated human WI-38 fibroblasts. This molecule was found to enhance growth rate and cell densities in actively dividing cultures. As the cells aged, however, this stimulatory effect diminished and finally vanished. In a long term experiment the enhancement of growth of procaine treated cultures was finally replaced by a toxic effect even at low concentration. The amount of the thermolabile enzyme found in phase III cells did not change when procaine was added to the culture medium. In this cellular aging model, procaine behaved like a metabolic stimulator of actively dividing cells but not as an "antiaging" molecule as it is sometimes assumed.

Cell Division↗
PubMed1987

Alteration of the microtubule organization in aging WI-38 fibroblasts. A comparative study with embryonic hamster lung fibroblasts.

The microtubule organization in human WI-38 fibroblasts subcultivated in vitro has been investigated using nocodazole, a reversible inhibitor of the microtubules. Two phenotypes were observed. The typical fibroblast cells, called Type 1 cells, showed, after nocodazole treatment, a centripetal depolymerization wave of the microtubules and the giant Type 2 cells which have a more heterogeneous behaviour. Some of the cells clearly showed a centrifugal depolymerization of the microtubules, others a mixed behavior and less than 1% displayed the same behavior as the Type 1 cells. Confirming previous data obtained with Hamster fibroblasts (Raes et al., 1983, 1984), these results suggest a modification in the microtubule organization which could account for the aberrant division of some WI-38 cells in aged cultures. The relevance of this observation for the emergence of the morphologically different Type 2 cells and for cell division impairment in serially in vitro cultivated cells is discussed.

Aging↗
PubMed1987

Comparative study of the enzymatic defense systems against oxygen-derived free radicals: the key role of glutathione peroxidase.

Human WI-38 diploid fibroblasts have been cultivated under high toxic O2 pressure, and their survival curves are reported. Superoxide dismutase, catalase, or glutathione peroxidase provided some protection when injected in the cells exposed to O2. This protective effect, recorded after 3 or 4 days of incubation, was the most pronounced when cells were injected just before oxygen exposure. Quantitative injection assays have been performed for the three enzymes. Surprisingly, glutathione peroxidase was found to be much more effective than both catalase and superoxide dismutase, the latter being particularly inefficient.

Catalase↗