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K Decker

Publications and source records attributed to K Decker.

At least 19 recordsLinked to original sources
PubMed1992

[Tumor necrosis factor alpha].

TNF-alpha (Cachectin) is a cytokine (tissue hormone) synthesized and secreted as a trimeric polypeptide; major producers are the cells of the RES after stimulation, e.g. by endotoxin. Its synthesis is strictly regulated. TNF elicits a great number of cell-specific responses. IL-1 and probably additional mediators cooperate in these effects. TNF is instrumental not only in the pathogenesis of inflammation, infections and some cell injuries, but also in the unspecific immune response, tumor toxicity and tissue homeostasis. TNF binding to specific receptors on cell surfaces initiates of secondary intracellular signals that, in turn, mediate the metabolic responses of the target cells. Pharmacological intervention may be envisaged at the level of synthesis (glucocorticoids, anti-oxidants) or interaction of the cytokine with its receptors.

Humans↗
PubMed1992

Eicosanoid-mediated contractility of hepatic stellate cells.

To approach experimentally the problem of contractility, stellate cells from rats were isolated and grown on a flexible silicone rubber substrate. Increases or decreases in the number of wrinkles of the silicone membrane beneath the cells that were easily observable by microscopy was employed as semi-quantitative measure of stellate cell motility. Contraction of stellate cells accompanied by diminution of cell body size was induced by U46619 (a thromboxane A2 analogue) and prostaglandin (PG) F2 alpha. Wrinkle formation became detectable 1.5 min after addition of 2 microM-U46619 and reached its maximum 10-15 min later. The effect of PGF2 alpha was not so striking, but lasted for a longer period of time. On the other hand, dibutyryl cyclic AMP, Iloprost (a PGI2 analogue) and PGE2 led to the disappearance or decrease in the number of wrinkles, indicating relaxation of contracted stellate cells. For instance, after addition of 2 microM-Iloprost, 47, 75 and 82% of contracted stellate cells had relaxed within 5, 10 and 20 min respectively. Moreover, dibutyryl cyclic AMP induced disappearance of alpha-smooth muscle actin stress fibres. This response became recognizable 10 min after addition of dibutyryl cyclic AMP; 40 min later, 97% of stellate cells were devoid of stress fibres. Thus stellate cells are able to undergo reversible contraction in primary culture, and the contraction of these cells may be mediated by eicosanoids that can be produced within the liver.

Animals↗
PubMed1992

Proteinkinase C beta-isoform triggers the formation of prostanoids and superoxide in liver macrophages.

The zymosan- and phorbolester-induced formation of prostanoids in cultured rat liver macrophages has been shown recently to be controlled by proteinkinase C (1). Using specific antibodies raised against the alpha-, beta-, gamma- and epsilon-isoforms of proteinkinase C, we show that proteinkinase-beta is the predominant isoform in rat liver macrophages. Northern blot analysis with a beta-isoform-specific c-DNA probe revealed the expression of m-RNA for proteinkinase-beta. In resting cells the beta-isoform of proteinkinase C is nearly equally distributed between the cytosolic and membrane fractions. Zymosan and phorbolester led to a translocation of proteinkinase-beta from the cytosol to the membranes, whereas exogenously added arachidonic acid and the calcium ionophore A23187 had no effect. These data indicate that the beta-isoform of proteinkinase C takes part in the prostaglandin and superoxide formation following PMA and zymosan treatment of rat liver macrophages.

Animals↗
PubMed1992

[Fatal course of peracute fatty liver of pregnancy].

A fatal case of acute fatty liver of pregnancy (AFLP) is reported. After admission, the patient was delivered within 3 hours. Routine laboratory investigation revealed acute liver insufficiency with advanced coagulopathy. Despite substitution therapy, the severe coagulation defect progressed to lethal intracerebral bleeding. Advanced AFLP can only be satisfactorily diagnosed in time, if non-specific symptoms or icterus lead to studies of blood chemistry, especially liver function tests, coagulation parameters (including platelet count, fibrinogen, AT III), blood glucose and renal function (including uric acid). This will enable an adequate management of the patient. The clinical problem of AFLP still remains that of early diagnosis.

Acute Disease↗
PubMed1992

Rat tumor necrosis factor-alpha. Transcription in rat Kupffer cells and in vitro posttranslational processing based on a PCR-derived cDNA.

A DNA fragment with a reading frame of 708 basepairs coding for TNF-alpha from rat liver was cloned by the polymerase chain reaction. Using this species-specific cDNA a biotin-labelled antisense (-)RNA was transcribed. This probe was used for Northern blot analysis of TNF-alpha gene activation. Exposure of rat Kupffer cells to LPS led to a time-dependent change of TNF-alpha-mRNA expression with a maximum between one and two hours after stimulation. In vitro translation was carried out with sense (+)RNA in the presence of microsomes. SDS gel electrophoresis of the immunoprecipitated proteins revealed the formation of polypeptides with estimated molar masses of 26 and 17 kDa. They correspond to the estimated molar masses of a precursor of TNF-alpha encoded by the entire reading frame and of the mature form of TNF-alpha, respectively. A sequence of the cDNA assumed to code for the leader peptide was deleted and the remainder used for the construction of an expression plasmid. Using this construct, biologically active mature TNF-alpha was expressed in E. coli (10(6) units TNF-alpha per liter of culture medium). The propeptide as well as the biologically active TNF-alpha possess a homology of 92 and 76% to mouse and human TNF-alpha, respectively.

Amino Acid Sequence↗
PubMed1992

Prostaglandin D2 and E2 syntheses in rat Kupffer cells are antagonistically regulated by lipopolysaccharide and phorbol ester.

Prostaglandin-synthesizing activities were demonstrated in cell-free extracts of rat Kupffer cells and characterized. The enzymatic properties of PGH2 synthase were found to be similar to those of synthases present in other organs or cell types. The specific activity of the enzyme was not changed by substances that stimulate prostanoid release by intact Kupffer cells; however, it was reduced by pretreatment of the cells with glucocorticoid hormones. On the other hand, the activities of PGD2 and PGE2 synthase were influenced differently by the kind of cell stimulation. While pretreatment of the intact cells with endotoxin and/or inhibition of protein kinase C led to an enhanced PGE2 formation in cell-free extracts, exposure to agents that enhance protein kinase C-dependent signalling pathways, e.g. phagocytotic stimuli or phorbol ester, suppressed PGE2 synthase activity and, therefore, led to enhanced PGD2 synthesis. It is in line with this observation that in vitro activation of protein kinase C of Kupffer cells resulted in a reduced PGE2 and an enhanced PGD2 synthase activity.

Animals↗
PubMed1992

Regulation by prostaglandin E2 of cytokine-elicited nitric oxide synthesis in rat liver macrophages.

Nitric oxide (NO), apart from its properties as a vasodilator, is a cytotoxic agent released from macrophages upon stimulation with immunomodulating agents such as interferon-gamma and endotoxin. In rat Kupffer cells endotoxin causes the release of NO as well as of tumor necrosis factor-alpha and prostaglandin E2 (PGE2). This eicosanoid and its second messenger, cyclic AMP, have been shown to increase nitric oxide formation in Kupffer cells treated with endotoxin (Gaillard et al. (1991) Pathobiology 59, 280-283). But not only added PGE2 but also the prostaglandin produced endogenously upon stimulation with endotoxin increases NO synthesis. Neither tumor necrosis factor-alpha nor interleukin-1 beta stimulate NO synthesis by themselves, but together with PGE2 they are as effective as lipopolysaccharide plus PGE2. To replace PGE2 in the combination with the cytokines, however, dibutyryl cAMP has to be present in higher concentrations than with LPS. Interleukin-6 alone or in combination with PGE2 or dibutyryl cAMP is without any effect. Anti-TNF-alpha as well as anti-PGE2 antibodies reduce the release of NO upon stimulation with LPS. Consequently, the effect of LPS on NO production seems to be in part due to the self-stimulating effect of PGE2 and some cytokines, both produced by Kupffer cells upon LPS stimulation.

Animals↗
PubMed1992

How and why are some riboflavin coenzymes covalently attached to proteins?

Covalent flavinylation as elucidated by the formation of the histidyl(N3)-8 alpha-flavin bond in 6-hydroxy-D-nicotine oxidase of A. oxidans proceeds by an non-enzymatic mechanism. Incubation of the apoenzyme, FAD and a three-carbon phosphate ester at neutral pH leads to the formation of an enzymatically fully active holoenzyme. The role of His71 in this process was illustrated by site-directed mutagenesis. Nevertheless, the question whether covalent attachment of the cofactor in a holoenzyme has prevailed throughout the evolutionary screening process because of biological significance or whether it is a chance event of neutral selective value is still open.

Animals↗
PubMed1991

Ca(2+)-induced reversible translocation of phospholipase A2 between the cytosol and the membrane fraction of rat liver macrophages.

In cell-free extracts of rat liver macrophages (Kupffer cells) phospholipase A2 was found to be rapidly associated with the particulate fraction in a Ca(2+)-dependent manner at Ca2+ concentrations of 0.1-1.0 microM. This is also the range of the levels of intracellular Ca2+ reported for basal and various stimulated conditions. After translocation, phospholipase A2 could be released from the membranes in the presence of Ca2+ chelators, increasing the specific activity of phospholipase A2 in the supernatant fraction. These findings support the view that translocation is a regulatory mechanism of phospholipase A2 by bringing the enzyme to its substrate. Unlike the situation with protein kinase C, Mg2+ exerted little effect on phospholipase A2 translocation, indicating that this process is regulated in vivo mainly by fluctuations of the intracellular Ca2+ content.

Animals↗
PubMed1991

Synergistic effect of magnesium and calcium ions in the activation of phospholipase A2 of liver macrophages.

In cell-free extracts of rat liver macrophages (Kupffer cells) phospholipase A2 was found to be strongly activated at free Ca2+ concentrations from 100 nM to 1 microM in the presence of 4 mM free Mg2+. This is within the range of intracellular free Ca2+ reported for basal and various stimulated conditions, respectively. Ca2+ alone increased phospholipase A2 activity at high Ca2+ concentrations (1 mM) whereas Mg2+ alone had only little stimulatory effect. Calmodulin does not seem to participate in the regulation of phospholipase A2 although it relieved the inhibition of phospholipase A2 activity by calmodulin antagonists.

Animals↗
PubMed1991

Regulation of nitric oxide production by stimulated rat Kupffer cells.

Macrophages have been described to release nitric oxide (NO) as a cytotoxic radical. This highly unstable substance is as well known as endothelium-derived relaxing factor produced by vascular endothelial cells. Because of its cytotoxic activity the synthesis of NO by rat Kupffer cells, the liver macrophages, upon stimulation with endotoxin (lipopolysaccharide; LPS) and tumor necrosis factor-alpha (TNF-alpha) in combination with prostaglandin E2 (PGE2) and dibutyryl cAMP (dBcAMP) was studied. Kupffer cells were stimulated after 48 h of primary culture. NO was quantified as NO2- in the cell medium 24 h after stimulation. LPS stimulated NO generation 5- to 10-fold over the basal level. This increase could be further enhanced by PGE2 and dBcAMP, especially when added 1 h after LPS. NO generation after stimulation with LPS or LPS + PGE2 depended on the simultaneous production of PGE2 by the stimulated Kupffer cells. It could be partly inhibited by anti-PGE2 antibody or acetylsalicylic acid. While murine TNF-alpha did not stimulate NO synthesis significantly, added PGE2 raised NO synthesis about 6-fold. The addition of dBcAMP to TNF-alpha in the same concentration as with LPS, however, had no effect. Thus, stimulation by LPS + PGE2 equals that of LPS + dBcAMP whereas TNF-alpha + PGE2 does not equal TNF-alpha + dBcAMP, indicating differences in the mode of action of PGE2 on LPS- or TNF-alpha-treated Kupffer cells.

Animals↗
PubMed1991

Endotoxin-refractory liver macrophages secrete tumor necrosis factor-alpha upon viral infection.

Rat liver macrophages (Kupffer cells) secrete tumor necrosis factor-alpha (cachectin) after exposure to Newcastle disease virus or bacterial endotoxin. Macrophages treated with endotoxin become refractory and fail to release tumor necrosis factor-alpha to a secondary challenge with endotoxin. The acquisition of the refractory state is dose-dependent, requires the continuous presence of endotoxin for a minimum of 8 h, is transient, and reversible. Endotoxin, however, renders Kupffer cells unresponsive only to itself. When endotoxin-refractory macrophages are activated by Newcastle disease virus, they still secrete tumor necrosis factor-alpha in amounts expected with this stimulus. Immunoprecipitation studies show that the precursor of tumor necrosis factor-alpha is found only in lysates of endotoxin-sensitive, but not in refractory macrophages, thus arguing against a post-translational regulatory process. Whereas prostaglandin E2 inhibits the production of tumor necrosis factor-alpha in response to endotoxin and viruses, it does not appear to mediate the refractory state.

Animals↗
PubMed1991

Signal transduction in endotoxin-stimulated synthesis of TNF-alpha and prostaglandin E2 by rat Kupffer cells. Role of extracellular calcium ions and protein kinase C.

Endotoxin is a well established elicitor of cytokine production in mononuclear cells. Nevertheless, the path of signal transduction between the crucial contact of the cells with endotoxin (lipopolysaccharide) and the synthesis and release of the mediators is yet poorly understood. In particular, the involvement of Ca2+ and protein kinase C in this process is still a matter of controversy. Here, it will be demonstrated that removal of extracellular Ca2+ by EGTA does not have a significant effect on the endotoxin-stimulated production of tumor necrosis factor-alpha (TNF-alpha) and on total protein synthesis in rat Kupffer cells. However, the release of prostaglandin E2 could not be raised above the basal level under these conditions. Treatment with inhibitors of protein kinase C such as the isoquinoline derivative, H-7, or staurosporin is without influence on TNF-alpha synthesis. The depletion of protein kinase C through preincubation of rat Kupffer cells with phorbol 12-myristate 13-acetate for 24 h was also without effect on TNF-alpha production. The effectiveness of these inhibitors under the conditions used was ascertained by measurement of the O2- release from the same cell batches. Superoxide production known as protein kinase C-dependent in Kupffer cells (Dieter et al. (1986) Eur. J. Biochem. 86, 451-457) was suppressed in a dose-dependent manner by staurosporin or after prolonged pretreatment with the phorbol ester. H-7 decreased superoxide production only slightly in high doses that severely harm the Kupffer cells. Prostaglandin E2 release, although clearly protein-kinase C-dependent in phagocytosing rat Kupffer cells, is not decreased following exposure to lipopolysaccharide in the presence of protein kinase C inhibitors.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1991

Fate of exogenous and endogenous prostaglandins D2 and E2 in the perfused rat liver.

The degradation of radiolabelled exogenous PGD2 and PGE2 was compared to that of endogenous labelled prostaglandins synthesized after stimulation with PMA in the perfused rat liver. With exogenous PGD2 and PGE2 the same degradation products were found in the perfused liver as in hepatocyte primary cultures. The major metabolite of PGD2 was dinor-PGD2 while tetranor-PGE1 was the main degradation product of PGE2. Some polar metabolites and tritiated water were also formed. The metabolites detected with endogenous prostaglandins were similar to those obtained with exogenous PGD2. Over 99% of the labelled prostaglandins were degraded in the recirculating perfusion within 40 min. In the open perfusion system, 95% of endogenous PGD2 was calculated to be degraded after a single passage through the liver, which suggests that hepatocytes play an important role in the removal of biologically active prostaglandins.

Animals↗
PubMed1990

Activation of GD3 synthase by sex steroid hormones in cultured rat hepatocytes.

The influence of sex steroid hormones on the activities of GM3 and GD3 synthases in isolated hepatocytes was studied. Progesterone (0.1 - 2.0 microM), beta-estradiol (0.1 - 1 microM), and testosterone (0.1 - 1 microM) activate GD3 but not GM3 synthase when added directly to hepatocytes cultured in modified William's E medium.

Animals↗