[Nephrotic syndrome. Development, complications and treatment].
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Biomedical subjects
Publications and source records attributed to U Sester.
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Immunologic complications of chronic renal failure are associated with the overproduction of proinflammatory cytokines by monocytes. This is partly due to renal failure itself but is further enhanced by hemodialysis treatment with frequent contact between blood and dialyzer membranes. Previous studies have shown an imbalance of proinflammatory and regulatory monokines in these patients. This study examines monokine production in hemodialysis patients using for the first time a very sensitive method of cytokine detection at a single-cell level by flow cytometry ("cytoflow technique"). Monocytes were stained intracellularly for the production of interleukin-6 (IL-6) and IL-10 after 20 h of culture with lipopolysaccharide. It was shown that high levels of proinflammatory IL-6 in hemodialysis patients are due to an increased number of monocytes producing this cytokine, while IL-6 synthesis per cell remains unchanged. In contrast, elevated levels of regulatory IL-10 are due to an increased synthesis per cell. This study demonstrates that in healthy subjects there is a population of monocytes producing exclusively IL-10 after 20 h of stimulation by lipopolysaccharide. This distinct population of regulatory monocytes is infrequent in dialysis patients, in whom most of the IL-10-positive monocytes also produce IL-6. These findings indicate that overproduction of proinflammatory factors in dialysis patients is at least in part due to a loss of cytokine-specific differentiation in monocytes.
1. The patient, a 59 year old woman, developed a state of acute excitation several hours after the administration of 400 mg of the fluorquinolone pefloxacin in combination with 1000 mg paracetamol. Nine days later, after a total dosage of pefloxacin of 800 mg, she was admitted to our hospital with a psychotic disorder. 2. There was a full remission of symptoms after treatment with perazine up to a dosage of 500 mg/day. 3. Three years ago, the patient had developed a manic state under a medication with corticosteroids. 4. So far, the mechanism of--in this case--long-lasting central nervous side effects of fluorquinolones is not known. In patients with increased vulnerability of the CNS or in advanced age the application of fluorquinolones should be considered critically, in particular in combination with non-steroidal anti-inflammatory drugs.
Superoxide radical (SOR) formation in the brain and the liver of male Chinook salmon (Oncorhynchus tschawytscha) increased in mitochondrial and plasma membrane samples as they aged. In 2-year-old salmon, spawning also lead to a significant elevation in SOR formation in mitochondrial and plasma membrane samples. The rise in this free radical was associated with an increase in lipid peroxidation, a decrease in plasma membrane fluidity, and an elevation in cathepsin B activity in the brain and liver. In 2-year-old spawning salmon, the changes in these parameters was greater than in 2-year-old non-spawning salmon. These observations suggest that free radical levels increase with aging and during spawning and indicate that these changes may be involved in cellular degeneration. In addition, these results support the suggestion that cellular degeneration accelerates during the spawning process.
Plasma membrane samples from rat brain, heart, and liver were examined for biochemical changes with age. A rise in superoxide radical (SOR) levels was followed by increases in thiobarbituric acid reactive substances and decreases in membrane fluidity with age. The earliest rise in SOR formation appeared in the plasma membrane from the brain. With age, protein synthesis also decreased significantly in tissue homogenates from brain and heart but was unchanged in the liver. Exposure of plasma membrane samples to in vitro-elevated SOR levels stimulated formation of lipid peroxides, as indicated by the thiobarbituric acid test, and resulted in a decrease in membrane fluidity in each tissue and in a decline in protein synthesis in brain and heart. Changes in brain lipid peroxidation and in membrane fluidity in brain and heart as a result of SOR supplementation were further enhanced due to age. In addition, the mechanism of SOR formation was examined in plasma membrane samples from the brain. SOR generation was Ca(2+)-sensitive, blocked by superoxide dismutase or vitamin E and inhibited by both indomethacin, a cyclooxygenase inhibitor, and bromophenacyl bromide, a phospholipase A2 inhibitor. These results show significant increases in SOR formation and biochemical alterations in plasma membranes from brain, heart, and liver in aging rats. SOR formation appears to be enzyme-mediated and elevated levels of this oxygen radical could be involved in membrane breakdown in older rats.
The nuclear protein statin, detectable with specific monoclonal antibodies, is found mostly in nonproliferating cells (Wang, E. (1985) J. Cell Biol. 100, 545-551). In the rat liver a 57-kDa protein designated as rat liver protein 57 (RLp57) was recently identified to carry the epitope for the anti-statin-specific monoclonal antibody, S-44 (Sester, U., Moutsatsos, I. K., and Wang, E. (1989) Exp. Cell Res. 182, 550-558). To characterize further the RLp57 protein, in the present study a polyclonal antibody was raised to the RLp57 protein eluted from polyacrylamide gel. Similar to the anti-statin monoclonal antibody, this polyclonal antibody recognizes a nuclear antigen in nonproliferating fibroblasts and reacts with a 57-kDa protein in rat liver and nonproliferating cells strongly suggesting that RLp57 is a statin protein from rat liver. Two isoforms of RLp57 (isoelectric points between 6.5 and 7.0) were detected after two-dimensional gel electrophoresis and immunoblotting. RLp57 was purified using multiple chromatographic steps, including ion-exchange and affinity chromatography followed by chromatofocusing. These results show that RLp57, a statin protein found in liver, has two isoelectric variants and can be purified to apparent homogeneity by sequential steps of chromatographic procedures.
The nuclear protein statin, found uniquely in nonproliferating cells, has been previously identified by mouse monoclonal antibodies designated as S-30 and S-44. We report here the screening of various rat tissues for proteins cross-reacting with these antibodies. As revealed by immunoblotting, two polypeptides (mol wt 80 and 57 kDa) and a group of lower-molecular-weight proteins migrating between 34 and 38 kDa were found to react with the anti-statin antibodies. The most prominent immunoreactivity was observed with a 57-kDa protein present in rat liver. Upon further fractionation of the liver protein extract with ammonium sulfate [(NH4)2SO4] the 57-kDa protein, designated as rat liver protein 57 or RLp57, was detected independently with both anti-statin antibodies in the 30 to 60% (NH4)2SO4 fraction. In order to determine whether rat liver protein 57 is indeed specifically recognized by anti-statin antibody S-44, we used RLp57 transferred onto nitrocellulose paper as a specific substrate for the adsorption of the S-44 antibody from ascites fluid. As shown by indirect immunofluorescence microscopy the solution remaining after adsorption failed to stain human fibroblasts. The adsorbed immunoglobulin, however, upon elution revealed statin-specific nuclear staining activity on senescent fibroblasts. These findings suggest that rat liver protein 57 and the human fibroblast statin share similar antigenic determinants recognized by the statin-specific S-44 antibody. Our results indicate furthermore that the statin previously identified in fibroblasts may represent one member of a group of several antigenically related proteins detectable with specific anti-statin antibodies.
The ultrastructure of synapses in the stratum fibrosum marginale (SM) of the optic tectum of cichlid fishes (Tilapia mariae) was investigated following acclimation to extreme environmental temperatures (experimental temperatures 16 degrees C and 32 degrees C; preference temperature: 26 degrees C). Four different types of synapses can be differentiated, from which 90% of all synapses belong to axodendritic spine synapses. The general ultrastructural appearance is more distinct in case of cold -adapted fishes, which in particular is due to a more pronounced staining of the polysaccharide-surface coat of the membranes. Analyzing quantitatively the axo-dendritic spine synapses (600 of each collective) significant differences were found for the length of the synaptic contact zone (16 degrees C: 0,391 micron; 32 degrees C: 0,369 micron) and for the thickness of postsynaptic densities (16 degrees C 42,6 nm; 32 degrees C: 52,7 nm). The number of mitochondria per presynapse was rised for cold-adapted animals to a value of 0,29 as compared to 0,18 mitochondria/presynapse for warm-adapted animals. The vesicle density per micron 2 of synaptic area, however, remained unchanged (approximately equal to 89 vesicles/micron 2). The morphological synaptic plasticity thus demonstrated in the optic tectum of chichlid teleosts, is correlated with behavioural changes and with alterations in the biochemical composition of synaptic membranes.
Drosophila were examined to see if there is an increase in damage associated with free radical activity in older flies. The levels of superoxide radical and of lipid peroxides were higher in membrane samples from older flies. Also, in older Drosophila there was a significant decrease in membrane fluidity, as determined by fluorescence polarization, and an increase in ATP-dependent calcium uptake. In homogenates from whole flies, the concentration of inorganic peroxides and activity of the proteolytic enzyme, cathepsin B, increased with age. During their lifetime, vestigial wing Drosophila displayed a greater level of free radical activity than wild-type flies and a significantly shorter life span (26.7 +/- 0.7 days vs. 34.4 +/- 1.0, p < 0.01). These results indicate that the level of oxidative stress is closely coupled to cellular damage and to life span, and they indicate that free radicals may play a central role in the aging process in Drosophila.