PubMed Health⌕ Search

Biomedical subjects

W Engels

Publications and source records attributed to W Engels.

At least 37 records · Page 2Linked to original sources
PubMed1993

Rapid and highly sensitive high-performance liquid chromatographic method for the determination of histamine and 3-methylhistamine in biological samples using fluorescamine as the derivatizing agent.

A highly sensitive and rapid high-performance liquid chromatographic assay for the determination of histamine and 3-methylhistamine in biological samples using 1-methylhistamine as the internal standard is described. Samples were purified and concentrated on cation-exchange columns and derivatized with fluorescamine. The lower detection limit was 20 pg on-column. Linearity was demonstrated up to 20 ng on-column. The samples could be derivatized simultaneously before injection and were stable for 7 days. The method was used for the determination of histamine and related compounds in coronary perfusates, extracts of homogenized rat hearts, and supernatants of stimulated peritoneal mast cells.

Animals↗
PubMed1993

Formation of prostanoids and hydroxy fatty acids by stimulated peritoneal mast cells: role of the dietary fat type in rat.

To study the influence of membrane fatty acid composition on the formation of prostanoids and hydroxy fatty acids by rat peritoneal mast cells (MC), animals were fed three different types of fatty acids: mackerel oil (MO), abundant in n-3 fatty acids; sunflower seed oil (SO), rich in linoleic acid; and hydrogenated coconut oil (HCO), mainly containing saturated fatty acids. The presence of n-3 fatty acids in the diet resulted in the incorporation of 20:5(n-3), 22:5(n-3) and 22:6(n-3) in MC phospholipids. A decrease of arachidonic acid, 20:4(n-6), was observed in MC-phospholipids of the MO-fed animals. Furthermore, increasing the relative amounts of 18:2(n-6) in the diet (SO group) led to an increased incorporation of linoleic acid, 18:2(n-6) in MC phospholipids when compared to both other dietary groups. The changes in MC phospholipid fatty acid composition were (partly) reflected in the formation of prostanoids and hydroxy fatty acids upon stimulation with the calcium ionophore A23187. The decrease in arachidonic acid content in MC phospholipids of MO-fed rats resulted in a decreased formation of PGD2 when compared to both other groups. Also, the increased amounts of 18:2(n-6) in MC phospholipids of SO-fed rats resulted in an increased formation of 9- and 13-HODE upon stimulation. The results show that modifications in the fatty acid composition of the diet influences MC membrane fatty acid composition which ultimately results in changes in prostanoid and hydroxy fatty acid synthesis by MC upon stimulation with the calcium ionophore A23187.

Animals↗
PubMed1993

DNA homology requirements for mitotic gap repair in Drosophila.

We used P transposable-element mobilization to study the repair of double-strand DNA breaks in Drosophila melanogaster premeiotic germ cells. Distribution of conversion tracts was found to be largely unaffected by changes in the length of sequence homology between the broken ends and the template, suggesting that only a short match is required. However, the frequency of repair was highly sensitive to single-base mismatches within the homologous region, ranging from 19% reversion when there were no mismatches to 5% when 15 mismatches were present over a 3455-bp span.

Alleles↗
PubMed1993

Production of arachidonic acid metabolites in adult rat cardiac myocytes, endothelial cells, and fibroblast-like cells.

Cells were incubated in the presence of the Ca2+ ionophore A23187 (10 microM) and arachidonic acid (AA, 80 microM). The release of eicosanoids from subcultivated cardiac endothelial and fibroblast-like cells amounted to 23.3 +/- 4.5 and 2.0 +/- 0.4 nmol/mg cellular protein per 30 min, respectively. The release from isolated cardiomyocytes remained below the detection limit of the high-performance liquid chromatography assay (< 0.00015 nmol/assay). When a very sensitive radioimmunoassay was applied, cardiomyocytes released 0.002 +/- 0.0001 nmol prostacyclin per milligram cellular protein per 30 min. Prostaglandin (PG) E2 and PGF2 alpha, 12-hydroxyheptadecatrienoic acid, 11- and 15-hydroxyeicosatetraenoic acid, and thromboxane B2 were the main eicosanoids released by endothelial cells. The stable product of prostacyclin, 6-keto-PGF1 alpha, contributed relatively little to the total amount of eicosanoids formed by endothelial cells. Fibroblast-like cells released predominantly PGE2 and 6-keto-PGF1 alpha and, to a lesser extent, 12-hydroxyheptadecatrienoic and 15-hydroxyeicosatetraenoic acids. Neither endothelial cells nor fibroblast-like cells released leukotrienes. A23187 stimulated eicosanoid release from endothelial cells when exogenous AA was below 40 microM. Addition of albumin reduced the amount of eicosanoids produced. Histamine and bradykinin did not influence 6-keto-PGF1 alpha and PGE2 production in cardiomyocytes. Histamine only gave rise to a slight but significantly higher release of 6-keto-PGF1 alpha in endothelial cells.

6-Ketoprostaglandin F1 alpha↗
PubMed1992

Effects of butylated hydroxyanisole on arachidonic acid and linoleic acid metabolism in relation to gastrointestinal cell proliferation in the rat.

In order to determine the effect of oral administration of 2(3)-tert-butyl-4-hydroxyanisole (BHA; dose-level: 1.5% BHA of the diet) on arachidonic acid (AA) and linoleic acid (LA) metabolism in correlation with changes in gastrointestinal cell kinetics, we coadministered two inhibitors of prostaglandin H synthase, acetylsalicylic acid (ASA) and indomethacin (IM), to rats. Coadministration of ASA (0.2%) and IM (0.002%) in the drinking water, resulted in a significant reduction of the BHA-induced enhancement of cell proliferation in forestomach and glandular stomach. ASA completely counteracted the effect of BHA on labeling indices in colon/rectum whereas IM exhibited no effect in this organ. Both inhibitors had no direct effect on cell kinetics in the control groups. ASA, and to a lesser degree IM, inhibited prostaglandin E2 release in all tissues examined. Whereas ASA did inhibit lipoxygenase-mediated metabolism of AA in forestomach tissue, ASA did not affect the release of AA- and LA-derived hydroxy fatty acids in glandular stomach and colon/rectum. IM did not affect lipoxygenase production. BHA, however, appeared to be a strong inhibitor of both routes of AA metabolism. While ASA nor IM affected LA metabolism, BHA inhibited both prostaglandin H synthase-mediated and lipoxygenase-mediated metabolism of AA and LA. A causal role of AA or LA metabolites in the process of cell proliferation enhancement induced by BHA, can therefore be excluded. Prostaglandin H synthase may, however, be involved in BHA activation by converting the hydroquinone metabolite of BHA to the corresponding quinone by redox cycling, which is probably accompanied by reactive intermediate production.

Animals↗
PubMed1992

The effect of viral infection on eicosanoid formation and procoagulant activity of rat peritoneal macrophages. Role of the dietary fat type.

The effect of dietary manipulation on eicosanoid formation in rat peritoneal macrophages was studied in relation to some of their effector functions: cellular procoagulant activity, production of reactive oxygen species (measured as chemiluminescence), and phagocytosis of antibody-coated erythrocytes. Rats were fed adequate diets for eight weeks containing mackerel oil (MO), sunflowerseed oil (SO) or hydrogenated coconut oil (HCO). The release of eicosanoids from resident macrophages stimulated by opsonized zymosan was significantly lower for the MO group as compared to the other dietary groups. Infection of the animals via intraperitoneal injection with rat cytomegalovirus resulted in a significant decrease in eicosanoid production in all groups, irrespective of dietary fat type. However, in the HCO group a partial restoration of TXB2 and HHT production could be observed at day 10 post infection. Resident macrophages obtained from the mackerel oil fed animals showed a significantly higher procoagulant activity than those from the other diet groups. Infection of the animals resulted in an increase in procoagulant activity in all groups. In contrast, no significant differences in chemiluminescence and in phagocytosis were detected between macrophages obtained from rats fed the different diet groups. It is concluded that peritoneal macrophages obtained from mackerel oil fed rats produce less eicosanoids and are more procoagulant than those from the other dietary groups, but a viral infection eradicates these differences. Therefore, a correlation between eicosanoid formation and effector functions studied could not be established.

Animals↗
PubMed1990

Ischemia and reperfusion induced formation of eicosanoids in isolated rat hearts.

Isolated, ejecting rat hearts, perfused with Krebs-Henseleit buffer, were exposed to various periods of global ischemia. Arachidonic acid (AA) accumulated significantly in the ischemic heart when the duration of ischemia exceeded 45 min. During 30 min of reperfusion, tissue levels of AA raised steadily to values of 10.5, 17.7, and 63.1 nmol/g, after 30, 45, and 60 min of ischemia, respectively. During reperfusion, significant amounts of AA metabolite prostacyclin (determined as stable metabolite 6-ketoprostaglandin F1 alpha, by radioimmunoassay and high-performance liquid chromatography) were released after 30, 45, and 60 min of ischemia. Beside prostacyclin, only small amounts of thromboxane B2 could be found during reperfusion. In contrast to increasing amounts of AA in reperfused tissue, prostacyclin release was maximal during the first 5 min of reperfusion and declined rapidly thereafter. Relatively small proportions of the accumulated AA are converted into prostacyclin, i.e., less than 1%. When hearts were treated with mepacrine, AA accumulation was almost completely abolished during 60 min of ischemia. The cumulative release of prostacyclin was found to be reduced to 134 pmol/g during 30 min of subsequent reperfusion. A close, rectilinear correlation could be established between AA accumulation and cumulative prostacyclin release during reperfusion. It is likely, however, that the site of bulk AA accumulation and that of conversion of AA into eicosanoids does not coincide in the ischemic and reperfused heart because of the low conversion rates of AA into prostacyclin and the different time courses of AA accumulation and prostacyclin production after reinstallation of flow.

Animals↗
PubMed1989

Influence of a cytomegalovirus infection on functions and arachidonic acid metabolism of rat peritoneal macrophages.

An intraperitoneal (rat) cytomegalovirus (RCMV) infection in the rat caused an influx of mononuclear cells, which have been altered in functions and arachidonic acid (AA) metabolism. Phagocytosis has been increased considerably 3 days postinfection (p.i.), whereas the release of prostacyclin, thromboxane A2, 12-hydroxyheptadecatrienoic acid (HHT), 5-hydroxyeicosatetraenoic acid (5-HETE), and leukotriene B4 (LTB4) was inhibited for more than 80%. The release of superoxide anions and the chemiluminescence response (CL) upon opsonized zymosan stimulation did not differ from those observed in resident peritoneal macrophages. Additionally, the levels of cyclic nucleotides (cAMP and cGMP) were low in both resident and influx macrophages (day 3 p.i.). In contrast, peritoneal macrophages harvested on day 10 p.i. still showed a high level of phagocytosis. However, the intracellular level of cyclic AMP had decreased fivefold, whereas CL response and superoxide anion release were inhibited significantly. Moreover, the production of prostacyclin, LTB4, and 5-HETE was still suppressed in contrast to thromboxane synthesis, which has selectively been restored in these macrophages. A direct regulatory role of AA metabolites in changes in macrophage functions that were due to a RCMV infection could not be demonstrated.

Animals↗
PubMed1988

Determination of prostaglandins and thromboxane in whole blood by high-performance liquid chromatography with fluorimetric detection.

A highly sensitive and specific assay for the quantitation of prostaglandins (PGs) such as PGE1, PGE2, PGF1 alpha, PGF2 alpha, 6-keto-PGF1 alpha, and including thromboxane B2, is described. The method involves the addition of PGF1 alpha and PGE1 as the internal standards, extraction from whole blood and purification by silica gel column chromatography. Following conversion into the methoximes, purification by reversed-phase chromatography and esterification with panacyl bromide, samples are analysed by high-performance liquid chromatography with fluorimetric detection. The lower limit of detection of the eicosanoids 6-keto-PGF1 alpha, thromboxane B2 and PGF2 alpha in blood is ca. 50 pg/ml and that of PGE2 is 100 pg/ml. Assay linearity is demonstrated over a range from 60 pg to 60 ng of eicosanoid injected. The method allows simultaneous assessment of prostaglandins and thromboxane extracted from complex biological fluids at picogram levels.

Chromatography, High Pressure Liquid↗
PubMed1987

Accumulation of lipids and lipid-intermediates in the heart during ischaemia.

The content of non-esterified fatty acids (NEFA) and their CoA and carnitine esters is low in normoxic cardiac tissue. The majority of fatty acids is esterified in the triacylglycerol and phosphoglyceride pool. During myocardial ischaemia beta-oxidation of fatty acids is inhibited. In addition, turnover of the esterified fatty acid pools is most likely disturbed. Accumulation of hydroxy fatty acids, acylCoA and acylcarnitine rapidly occurs after the onset of ischaemia. The accumulation of NEFA is a slower process. In addition to extracellular sources, NEFA originate also from intracellular lipid pools, most likely from phosphoglycerides. Although it has been suggested that activation of phospholipase A2 occurs in ischaemic tissue, the mechanism underlying the enhanced degradation of phosphoglycerides ist still incompletely understood.

Acyl Coenzyme A↗
PubMed1987

Influence of intracellular Ca2+-overload in eicosanoid synthesis of the myocardium.

Intracellular Ca2+-overload in the myocardium can be induced not only after readmission of Ca2+-containing fluid to rat hearts previously perfused with a Ca2+-free buffer, a phenomenon called "the calcium paradox", but also during administration of a Ca2+-ionophore to cardiac tissue. In rat hearts, the myocardial damage induced by the Ca2+ paradox was more pronounced than that after administration of the Ca2+-ionophore A23187, as indicated by the amount of lactate dehydrogenase released. The accumulation of NEFA, and especially arachidonic acid, was greater during the Ca2+ paradox than after the administration of the Ca2+-ionophore. Administration of the Ca2+-ionophore resulted in a considerable release of 6-keto-F1 alpha (the stable breakdown product of prostacylclin), and LTD4 and LTE4 (breakdown products of LTC4). In contrast, the formation of eicosanoids was absent during the Ca2+ paradox. It is concluded that the relation between Ca2+-overload and accumulation of arachidonic acid is ambiguous and that there is no close relation between the amount of arachidonic acid accumulated and the formation of eicosanoids in Ca2+-overloaded tissue. The absence of eicosanoid formation during the Ca2+ paradox might be explained by compartmentation of the arachidonic acid accumulation and its converting enzymes or impairment of the enzymatic machinery required for eicosanoid synthesis.

Animals↗
PubMed1987

Treatment of experimental cytomegalovirus infections with acyclovir.

Subcutaneous (s.c.) administration of Acyclovir (ACV) (100 mg/1 kg bodyweight) resulted in an ACV blood level of 80 microM at 2 h post infection. Thereafter the level declined rapidly reaching undetectable levels at 12 h post infection. Administration of pro-ACV (100 mg/kg body weight) by s.c. or intravenous (i.v.) route resulted in ACV levels of 75 microM and 160 microM respectively after 30 min. But here again the blood level of ACV declined rapidly and was completely disappeared after 12 h. Continuous administration of pro-ACV in daily doses of 100, 250 and 350 mg/kg body weight resulted in ACV blood levels of 15 microM, 19 microM and 39 microM, respectively. The effect of ACV and pro-ACV on the replication of CMV was measured in immunesuppressed rats. In rats inoculated with RCMV the daily administration of 25 to 50 mg ACV per kg body weight by s.c. injections twice daily, did not result in a reduction of virus titers in spleen and liver, but when the RCMV-infected rats were treated by 100 mg pro-ACV per kg body weight virus titers in the spleen and liver were significantly reduced as compared with those in sham-treated animals.

Acyclovir↗
PubMed1985

A quantitative method for assessing the third complement factor (C3) attached to the surface of opsonized Pseudomonas aeruginosa: interrelationship between C3 fixation, phagocytosis and complement consumption.

A direct enzyme-linked immunoassay is described for the determination of C3 that becomes attached to the surface of bacteria upon incubation in serum. The assay uses horseradish peroxidase-conjugated rabbit antiserum specific for human C3. In this study various Pseudomonas aeruginosa strains, Escherichia coli and Staphylococcus aureus were opsonized under different conditions and the amount of C3 fixed to the bacterial surface was measured directly. The extent of C3 fixation was compared with the percentage of phagocytosis by human polymorphonuclear leukocytes and with the complement consumption in the fluid phase. It was demonstrated that C3 fixation closely correlated with the percentage of phagocytosis but not with complement consumption. The method can be performed with relatively simple equipment and could be used in routine laboratories in order to determine the extent of opsonization of Pseudomonas aeruginosa and other bacteria and to detect opsonic defects in the sera of patients.

Complement Activation↗
PubMed1985

Role of lipopolysaccharide in opsonization and phagocytosis of Pseudomonas aeruginosa.

When the opsonization of various Pseudomonas aeruginosa strains--PAC 1, its O-chain-deficient mutant PAC 605, and an intermediate strain, P14--was measured either directly by determination of the amount of C3b attached to the bacterial surface or indirectly by assessing phagocytosis by human polymorphonuclear leukocytes and the responses of chemiluminescence, it was demonstrated that PAC 1 was opsonized and phagocytized to a lower extent than P14 and PAC 605. In contrast to PAC 605, PAC 1 showed an increased consumption of complement in the fluid phase and a rapid release of lipopolysaccharide antibodies bound to the bacterial surface due to the alternative pathway of the complement system. Furthermore, it was shown that with respect to PAC 1 and PAC 605, the lack of an O-chain resulted in increased sensitivity to serum and decreased virulence. From both in vivo and in vitro experiments, we concluded that the structure of the O-antigen polysaccharide chain of lipopolysaccharide is an important virulence factor of P. aeruginosa against the defense mechanisms of the host.

Complement Activation↗
PubMed1984

[A new treatment possibility for dogs with chronic or recurrent pyoderma].

Effective treatment of chronic or recurrent pyodermas in dogs is often found to be difficult. The disease apparently results from a change in the balance between the infecting microorganism: Staphylococcus aureus, and the host. This change in the normal equilibrium could be due to enhanced virulence of the infecting strain of staphylococci, but is more likely to have been caused by host factors such as disturbed non-specific defence mechanisms (of the skin) or malfunction of the immune response, e.g. hypersensitivity. When conventional forms of treatment such as grooming, skin-hygienic measures and antibiotics fail or relapses occur, immunotherapy with a staphylococcal vaccine or toxoid might be considered. In studies with experimental animals it was shown alpha-toxoid was superior to other staphylococcal toxoids or vaccines in preventing skin lesions. Four dogs with deep-seated chronic pyodermas were treated with staphylococcal alpha-toxoid emulsified in Freund's adjuvant. Three of these animals recovered, but because of severe unwanted side-effects, this form of therapy was abandoned. Ten dogs with deep-seated chronic primary pyodermas were treated with a commercially available alpha- and beta-toxoid preparation (Isopyos). Nine of these patients recovered. In two cases, however, relapses occurred several months later, but the symptoms disappeared again after a booster injection. One dog did not respond; its condition deteriorated and it had to be euthanised. As the dogs received toxoid intracutaneously as well as intramuscularly, the positive effect might be due to desensitization of the animals rather than to immunisation.

Animals↗
PubMed1984

Demonstration of an IgG-Fc receptor in ratcytomegalovirus infected cells.

Infection of rat embryonic fibroblasts (REF) and R2 cells with rat cytomegalovirus (RCMV) resulted in the formation of cytoplasmic and membrane receptors for the Fc region of immunoglobulin G. The Fc receptors were demonstrated by the immunocytochemical techniques using indirect fluorescence and peroxidase technique at the light microscope level and using the protein A adsorbed to colloidal gold technique for demonstration at the electron microscope level. The production level of demonstration at the electron microscope level. The production level of the Fc receptor in the cell was depending on the input of virus. Experiments with ara C, cycloheximide and actinomycin D revealed that the formation of the Fc receptor was dependent on RNA, DNA and protein synthesis. Furthermore, results indicate that redistribution of the Fc receptor to form a cap, takes place within the first 60 minutes.

Animals↗
PubMed1983

Activation of human prothrombin by stoichiometric levels of staphylocoagulase.

The activation of human prothrombin by the bacterial protein staphylocoagulase proceeds via the formation of a very stable equimolar complex. Unmasking of the active center in the prothrombin moiety of the complex is not caused by limited proteolysis. The kinetics of activation of human prothrombin by staphylocoagulase has been studied. The second order rate constant at pH 7.5, 37 degrees C, is 3.3 X 10(6) M-1 S-1. This reaction rate is close to reported diffusion-controlled rates of protein-protein interaction. The dissociation constant of the complex was too low to be measurable. From the kinetic data it is assumed that the first order rate constant for dissociation is orders of magnitude less than 10(-5) S-1. However, dissociation of the complex did occur in the presence of sodium dodecyl sulfate. Equimolar amounts of staphylocoagulase protect human thrombin, but not human factor Xa and bovine thrombin, against inactivation by antithrombin III. From these findings we postulate that tertiary structural changes in the thrombin region of prothrombin caused by a highly specific interaction between staphylocoagulase and that region unmask the active site.

Amino Acids↗