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W Engels

Publications and source records attributed to W Engels.

At least 19 recordsLinked to original sources
PubMed2006

Is nitric oxide involved in the tolerance of Calomys callosus as a reservoir host towards Trypanosoma cruzi infection?

Trypanosoma cruzi, the agent of Chagas disease, is known to cause enhanced nitric oxide (NO) production, which might be involved in host resistance. The inducible nitric-oxide-synthase (iNOS) is assumed to be responsible for the NO increase after several infections. We studied the potential role of NO in Calomys callosus, a natural reservoir of this protozoan parasite. The concentration of NO was determined in spleen and liver of animals infected with two different T. cruzi strains, BOL and BOL-SB. Furthermore, the iNOS mRNA expression was quantified in the same cell types. NO production was detectable in both tissues exhibiting only slight differences compared to non-infected controls. All measured NO values were significantly lower than those reported for a number of different mouse strains, which displayed extremely enhanced NO levels after T. cruzi infection. Surprisingly, iNOS mRNA expression was induced in infected C. callosus but without subsequent increase of NO levels, indicating a post-transcriptional regulation mechanism. In summary, our results, indicate that the tolerance of C. callosus to T. cruzi is only accompanied by non-toxic NO intracellular concentrations.

Animals↗
PubMed2004

Tyramine functions as a toxin in honey bee larvae during Varroa-transmitted infection by Melissococcus pluton.

From wounds of honey bee pupae, caused by the mite Varroa destructor, coccoid bacteria were isolated and identified as Melissococcus pluton. The bacterial isolate was grown anaerobically in sorbitol medium to produce a toxic compound that was purified on XAD columns, gelfiltration and preparative HPLC. The toxic agent was identified by GC-MS and FTICR-MS as tyramine. The toxicity of the isolated tyramine was tested by a novel mobility test using the protozoon Stylonychia lemnae. A concentration of 0.2 mg/ml led to immediate inhibition of mobility. In addition the toxicity was studied on honey bee larvae by feeding tyramine/water mixtures added to the larval jelly. The lethal dosis of tyramine on 4-5 days old bee larvae was determined as 0.3 mg/larvae when added as a volume of 20 microl to the larval food in brood cells. Several other biogenic amines, such as phenylethylamine, histamine, spermine, cadaverine, putrescine and trimethylamine, were tested as their hydrochloric salts for comparison and were found to be inhibitory in the Stylonychia mobility test at similar concentrations. A quantitative hemolysis test with human red blood cells revealed that tyramine and histamine showed the highest membranolytic activity, followed by the phenylethylamine, trimethylamine and spermine, while the linear diamines, cadaverine and putrescine, showed a significantly lower hemolysis when calculated on a molar amine basis. The results indicate that tyramine which is a characteristic amine produced by M. pluton in culture, is the causative agent of the observed toxic symptoms in bee larvae. Thus this disease, known as European foulbrood, is possibly an infection transmitted by the Varroa destructor mite.

Animals↗
PubMed2004

Six Trypanosoma cruzi strains characterized by specific gene expression patterns.

The intracellular parasite Trypanosoma cruzi, the causative agent of Chagas disease, is known to comprise heterogeneous populations. One possibility to explain the obviously distinct phenotypes of different T. cruzi strains is differential expression of particular genes. This could result in environmental adaptations of the parasite within host organs, leading to distinct clinical symptoms. With the aim of identifying differentially expressed genes, we examined different T. cruzi strains by suppression subtractive hybridization analysis. The isolated clones were sequenced and Blasted for sequence-homology with known T. cruzi genes. A stage-specific glycoprotein (82gp), an 85-kDa protein with homology to heat-shock proteins, a beta-tubulin gene, a hexosetransporter, a dehydrogenase/ prostaglandin F2alpha-synthase and a cathepsin B-like protease were identified. The expression of these genes was analyzed by RT-PCR. Diverse expression patterns were detected for different T. cruzi strains, but no specific correlation between the gene expression and the classification of groups could be found. We discuss the presumed importance of these T. cruzi gene expression patterns for future strategies of molecular therapy of Chagas disease. For pathological studies, other parameters such as distinct gene/antigen expression could also be of interest, because they probably likewise correlate with distinct phenotypes.

Animals↗
PubMed2003

Ultrastructure and bacterial infection of wounds in honey bee ( Apis mellifera) pupae punctured by Varroa mites.

The damage to western honey bee, Apis mellifera, colonies caused by the originally Asian ectoparasitic mite Varroa destructor is mainly a consequence of the infestation of host bee pupae. In the capped brood cell, female mites puncture the host's integument at preferred sites in order to suck haemolymph. Due to repeated feeding by the mother mite and her progeny, these perforations are kept open until shortly before the imaginal moult of the bee. Thereafter scarring takes place, thus preventing microbial infection after the adult bee has emerged from the protected environment of the sealed brood cell. However, colonies of various bacteria were found in the open wounds of about 15-30% of all inspected host pupae with an abundance depended on the level of host brood cell infestation by the mite. The small punctures of the pupal integument are difficult to detect but, by vital staining with trypan blue, the wounds can be visualised. The ultrastructure of the pupal wounds, the bacterial colonies and the scarring process are documented by a series of scanning electron micrographs.

Animals↗
PubMed2002

Molecular genetic characterization of different Trypanosoma cruzi strains and comparison of their development in Mus musculus and Calomys callosus.

Trypanosoma cruzi populations are characterized by diverse morphology, heterogeneous biological behavior, high genetic variability, and distinctly different clinical courses. The first objective of this work was to characterize different strains of T. cruzi with various molecular markers [simple-sequence-repeat PCR, randomly amplified polymorphic DNA (RAPD)-PCR, mini-exon genes]. All examined strains could be divided into two major lineages. Only one strain showed a different banding pattern in RAPD-PCR, which could be a further indication of the existence of a third lineage. The second aim was to examine the biological behavior of the different strains. Two animal models, Calomys callosus and Mus musculus, were infected. The results provide strong evidence that the biological behavior of the strains is not only lineage-specific. It appears that all factors, such as the infecting strain belonging to a certain lineage, the predominant morphological form of the isolate, and the immune response of the respective infected host, play an important role in the course of this infection.

Animals↗
PubMed1999

Microsatellite instability in Drosophila spellchecker1 (MutS homolog) mutants.

We have cloned a mutS homolog from Drosophila melanogaster called spellchecker1 (spel1) and have constructed spel1 mutant flies. MutS proteins promote the correction of DNA mismatches and serve important roles in DNA replication, recombination, and repair. The spel1 gene belongs to a subfamily of mutS first characterized by the MSH2 gene of yeast and which also includes hMSH2, one of the two major hereditary nonpolyposis colon cancer loci of humans. Like msh2 mutants in other species, we find that flies lacking the spel1 gene suffer a highly increased rate of instability in long runs of dinucleotide repeats when analyzed after 10-12 fly generations. Using a new assay, we have also discovered that mutations in spel1 decrease the stability of a dinucleotide repeat when it is copied into the site of a double-strand break during gene conversion. Contrary to the case in mammalian cells, spel1 deficiency does not affect tolerance of flies to a methylating agent nor does it affect resistance to gamma-irradiation.

Adenosine Triphosphatases↗
PubMed1999

Cytochrome P450, peroxisome proliferation, and cytoplasmic fatty acid-binding protein content in liver, heart and kidney of the diabetic rat.

Diabetes mellitus generally results in an increased systemic fatty acid mobilization which can be associated with an increase in mitochondrial and peroxisomal beta-oxidation of fatty acids in selected tissues. The latter is usually accompanied by a concomitant increase in the tissue content of cytoplasmic fatty acid-binding protein (FABP) which functions in the intracellular translocation of fatty acids. It was previously found that in liver clofibrate-induced proliferation of peroxisomes and increase in FABP expression each are dependent on the induction by cytochrome P4504A1 -mediated (CYP4A1) formation of dicarboxylic acids. We studied whether peroxisome proliferation and an increase of FABP contents in liver, heart and kidney of streptozotocin-induced diabetic rats are also accompanied by an increase of CYP4A1 activity, as this would indicate a possible regulatory role for dicarboxylic acids in peroxisome proliferation and FABP induction in diabetic organs other than liver. In livers of the diabetic rat, a concomitant increase was observed of the activities of CYP4A1 and the peroxisomal key enzyme fatty acyl-CoA oxidase (FACO) and of the FABP content. In the diabetic heart FACO activity and FABP content also increased, but there was no induction of CYP4A1 activity. Conversely, in diabetic kidney there was no increase in FACO activity nor FABP content in spite of a marked induction of CYP4A1 activity. It is concluded that streptozotocin-induced diabetes leads to increased peroxisome proliferation and increased levels of FABP in both liver and heart, which only in liver is accompanied by an induction of the cytochrome P450 system. Consequently, it is not likely that dicarboxylic acids are involved in the induction of peroxisome proliferation in the heart.

Acyl-CoA Dehydrogenase↗
PubMed1998

The role of mast cells and histamine in leukocyte-endothelium interactions in four rat strains.

The objective of the present study was to determine the role of mast cells and histamine in leukocyte-endothelium interactions in mesenteric venules of four rat strains: Brown Norway, Lewis, Sprague-Dawley and Wistar. Intravital microscopy showed that the mast cell stabilizer cromoglycate (5 mg/kg i.v. just before exteriorization of the mesentery) did not affect the baseline level and velocity of leukocyte rolling in any of the four strains. This finding is in agreement with the observation that cromoglycate pretreatment only slightly influenced mast cell degranulation in all strains except the Brown Norway. After mast cell stabilization, only in Sprague-Dawley did topical administration of histamine (10(-4) M) result in a significant increase in the level of leukocyte rolling and a decrease in the rolling velocity compared with the time control. Histamine induced leukocyte adhesion only in the Brown Norway strain. In conclusion, the hypothesis presented in other studies, that degranulation of mast cells, and more specifically the release of histamine, is of major importance for the induction of leukocyte-endothelium interactions in rat mesenteric venules is not generally applicable; the present study shows a clear strain dependency.

Animals↗
PubMed1997

Antigen-evoked mast cell degranulation in the isolated rat heart: no effect on subsequent ischemia-reperfusion induced damage.

In the present study, the possible role of mast cells in ischemia/reperfusion-induced myocardial injury was evaluated in the isolated 'mast cell depleted' rat heart. Hearts isolated from sensitized and non-sensitized rats were perfused according to Langendorff. After 30 min of normoxic perfusion, hearts were challenged with antigen, a procedure which is known to result in a massive mast cell degranulation in sensitized hearts. After another 20 min, both 'mast cell depleted' and control hearts were subjected to 30 min of ischemia followed by 30 min of reperfusion. The release of lactate dehydrogenase (LDH) was determined, to quantitate the extent of irreversible injury of cardiomyocytes. Histamine release was measured to establish mast cell degranulation. Coronary flow (CF) and left ventricular developed pressure (LVDP) were monitored to study the consequences of the procedures on hemodynamic recovery. It was found that both CF and LVDP significantly increased during the first min after antigen challenge. These changes were accompanied by an almost complete degranulation of cardiac mast cells. The increase in CF and LVDP values were rapidly followed by a decrease, reaching minimal values of 159 +/- 4% and 85 +/- 4% of those before administration of antigen, respectively, at 2-3 min after antigen challenge. No effect of antigen challenge on LDH release were found indicating that mast cell degranulation did not compromise myocyte integrity. During reperfusion following 30 min of ischemia both the increase in CF and LVDP in 'mast cell-depleted' hearts were not significantly different from those in control (non-sensitized) hearts. Similarly, at the end of the reperfusion-phase, CF and LVDP values in sensitized hearts were comparable to those in control hearts. Reperfusion results in increased LDH release, which at no point in time was significantly different between sensitized and non-sensitized hearts. In non-sensitized hearts histamine release during the reperfusion phase was not detectable. Therefore, the results indicate that in the isolated rat heart, mast cells are most likely not involved in acute ischemia/reperfusion-induced myocardial injury.

Animals↗
PubMed1997

Dietary modulation of fatty acid composition of mast cell phospholipids does not affect histamine release induced by compound 48/80.

OBJECTIVE AND DESIGN: In the present study we determined the extent to which the degranulation process in mast cells was related to the fatty acid composition of membrane phospholipids. MATERIAL: Peritoneal mast cells were isolated from Wistar rats (3 groups of 18 animals each), fed for 6 weeks diets which differed in their fatty acid compositions: (i) genuine salmon oil, abundant in (n-3) fatty acids, (ii) sunflower seed oil, rich in (n-6) fatty acids, particularly linoleic acid, and (iii) hydrogenated coconut oil, rich in saturated fatty acids. METHODS: Mast cells (10(6)/ml) were stimulated with various concentrations of the mast cell-degranulating agent, compound 48/80 (0.1-10 micrograms/ml). The extent of mast cell degranulation was quantified by determination of histamine in the supernatants using HPLC techniques. RESULTS: No differences in compound 48/80-induced histamine release between the three dietary groups for any of the concentrations of compound 48/80 tested were found. Analysis of variance followed by Tukey's method for multiple comparisons was used to evaluate the effect of changes in the dietary fat type. CONCLUSION: These findings strongly suggest that in contrast to the formation of eicosanoids, the process of mast cell degranulation by a receptor-independent pathway is not controlled by the fatty acid composition of membrane phospholipids.

Analysis of Variance↗
PubMed1997

Mast cell-mediated induction of ICAM-1, VCAM-1 and E-selectin in endothelial cells in vitro: constitutive release of inducing mediators but no effect of degranulation.

Mast cell (MC)-mediated induction of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) and of E-selectin was studied in cultures of rat heart endothelial cells (EC) and human umbilical vein EC (HUVEC) respectively. MC induced VCAM-1 and E-selectin, but hardly any ICAM-1 in EC. Induction was not dependent on MC degranulation, but seemed to be provoked by constitutively released substances, other than histamine, from MC. Co-incubation of MC and EC, allowing for direct contact between the two cell types, was more potent in induction than MC co-incubated separately from EC using a permeable membrane. MC were less potent in induction than exogenous added cytokines or LPS. Induction of cell adhesion molecules in rat heart EC was MC-specific, since EC incubations with either rat cardiomyocytes or heart fibroblasts had no effect. The data show that rat MC, independent of degranulation, secrete mediators relevant for the induction of a specific set of EC adhesion molecules in vitro. This suggests a (supportive) role for MC in cell-adhesion molecule induction in the endothelium in settings of early or mild inflammation. The results are discussed in the context of inflammatory processes in the heart in vivo.

Animals↗
PubMed1996

Lack of evidence for a role of mast cell degranulation in acute hypoxia/reoxygenation-induced injury in the isolated rat heart.

In the present study, we evaluated the potential role of mast cell degranulation in acute hypoxia/reoxygenation-induced injury to cardiomyocytes in the isolated rat heart. Histamine release was determined to delineate the extent of mast cell degranulation, whereas the release of creatine kinase (CK) and lactate dehydrogenase (LDH) was assessed to quantitate the extent of irreversible injury to cardiomyocytes. The suitability of peroxidase (PO) as a marker for mast cell degranulation was also evaluated. Reoxygenation resulted in a release of histamine corresponding with 6.5% +/- 0.6% of total tissue content, whereas LDH, CK and PO release amounted to 30% +/- 2%, 28% +/- 2% and 32% +/- 3% of their respective tissue contents. Identical perfusion in the presence of the mast cell stabilizer lodoxamide tromethamine resulted in a reduced histamine release (2.8% +/- 0.1%) of total tissue content upon reoxygenation, but the release of LDH, CK or PO was not influenced. Cumulative histamine release did not correlate with the amount of LDH, CK or PO released. Treatment with consecutive bolus injections of the mast cell degranulating compound 48/80 during normoxic perfusion resulted in an almost complete histamine release, whereas PO release remained below detection limit. When the compound 48/80-treated hearts were subjected to hypoxia/reoxygenation, the release of LDH, CK or PO during reoxygenation again remained unchanged, whereas histamine release was negligible. Determination of PO activity of freshly isolated cardiomyocytes demonstrated that the bulk of PO in rat hearts was located in this particular cell type. Therefore we conclude that in the isolated rat heart, PO release is not a specific marker of mast cell degranulation. In addition, our data provide no firm evidence that in this experimental model, mast cell degranulation contributes to a significant extent to acute hypoxia/reoxygenation-induced injury to cardiomyocytes.

Acute Disease↗
PubMed1996

Production and characterization of spontaneous rat heart endothelial cell lines.

Endothelial cells (EC) are important regulatory cells in physiology and pathology. in vitro studies with rat EC from heart tissue are hampered by laborious isolation and purification procedures, low yield, and limited lifespan of the cells. Therefore, it is essential to obtain long-term heart EC lines that offer a more adequate in vitro system for studying rat heart EC. An ex vivo perfusion model was used to isolate EC from rat heart. Isolation and culture conditions were modified to allow spontaneous development of immortalized rat heart EC (RHEC) lines. Produced cell lines were tested for endothelial nature using a panel of markers. The selected RHEC lines were subsequently tested for a series of phenotypic and functional properties representative of EC in the context of physiologic and inflammatory functions in vivo. A series of three spontaneous RHEC lines was produced from 13 isolations from Lewis rat hearts: RHEC-3, RHEC-10, and RHEC-11. These lines were stable for more than 30 passages (RHEC-3 for more than 100). The cell lines were tumorigenic and developed hemangiomas on in vivo injection. All three lines expressed major histocompatibility complex (MHC) class I but no MHC class II. Intercellular adhesion molecule 1 was only expressed by RHEC-3. Cytokine stimulation induced vascular cell adhesion molecule 1 in RHEC-3 and RHEC-11 as well as MHC class II in all three lines in different quantities. The phenotypic characteristics of the different RHEC lines resembled the myocardial microvascular endothelium in situ. The three lines expressed angiotensin-converting enzyme, and they responded to histamine and ATP but not to thrombin and bradykinin. They constitutively produced small amounts of endothelin and high levels of tissue plasminogen activator; they produced little (after stimulation with phorbol-ester PMA) or no von Willebrand factor. The RHEC lines produced thromboxane A2 but no prostacyclin; on stimulation with arachidonic acid and A23187, they produced prostaglandin E2. Therefore, we conclude the following. 1) The described isolation and culture technique is successful for production of spontaneous stable EC lines from rat heart. 2) RHEC-3, -10, and -11 can be considered a series of different lines representative of the heterogeneity of heart microvascular endothelium in vivo. 3) The RHEC lines offer a series of valuable tools to study various heart EC functions and mechanisms in physiology and pathology.

Animals↗
PubMed1995

Differential release of histamine and prostaglandin D2 in rat peritoneal mast cells: roles of cytosolic calcium and protein tyrosine kinases.

We studied how the release of histamine and prostaglandin D2 (PGD2) were connected in stimulated rat peritoneal mast cells, and to what extent these processes were controlled by the cytosolic Ca2+ concentration, [Ca2+]i, and protein tyrosine kinases. In the presence of 1 mM CaCl2, the G-protein activating compound 48/80 (10 micrograms/ml) evoked a transient rise in [Ca2+]i and a relatively high secretion of histamine, but only a low release of PGD2. In contrast, 5 microM thapsigargin (an inhibitor of endomembrane Ca(2+)-ATPases) and 5 microM ionomycin evoked high and prolonged rises in [Ca2+]i, and stimulated the cells to release relatively small amounts of histamine and high amounts of PGD2. Stimulation of the cells with CaCl2 and 10 microM ATP4- gave only minor quantities of histamine and PGD2, despite of the micromolar level of [Ca2+]i reached. When CaCl2 was replaced by EGTA, rises in [Ca2+]i as well as release of histamine and PGD2 were reduced with each agonist, but the preference of agonists to release more histamine or PGD2 remained unchanged. In mast cells with depleted Ca2+ stores, the addition of CaCl2 stimulated the store-regulated Ca2+ entry resulting in a prolonged rise in [Ca2+]i. However, simultaneous addition of compound 48/80 and CaCl2 was required for release of histamine and PGD2. In cells with full stores, PGD2 release evoked by compound 48/80 was greatly reduced by genistein and methyl-2,5-dihydroxycinnamate, two structurally unrelated inhibitors of protein tyrosine kinases, whereas histamine secretion was not influenced by these inhibitors. Similarly, with thapsigargin or ionomycin as agonist, PGD2 release was more sensitive to the tyrosine kinase inhibitors than histamine secretion. We conclude that in activated rat peritoneal mast cells: (i) the influx of extracellular Ca2+ potentiates agonist-evoked rises in [Ca2+]i as well as histamine secretion and PGD2 release; (ii) the amplitude of the [Ca2+]i rise does not determine the preferential effect of agonists to release more histamine or more PGD2; (iii) the relatively high PGD2 release evoked by thapsigargin and ionomycin is probably due to their potency to evoke a prolonged rise in [Ca2+]i and to activate protein tyrosine kinases.

Animals↗
PubMed1995

Transmural changes in mast cell density in rat heart after infarct induction in vivo.

The cardiac distribution of mast cells was investigated after the induction of acute myocardial infarction in the rat. The left anterior descending coronary artery (LAD) was occluded by ligation in the infarct group, whereas in sham rats only a superficial ligature was placed beside the LAD. Rats of both groups were killed at 4, 7, 14, 21, 35, and 85 days following surgery. Hearts were excised and formalin-fixed. Mast cell densities were monitored in subepicardial and subendocardial layers of the left ventricle (LV) in 6 microns thick toluidine blue-stained cross-sections. In control (non-operated) animals, mast cell densities were comparable in the LV subepicardial and subendocardial layers (1.5-2.0 cells per mm2). Following infarction, the mast cell density at the subepicardial site of the infarction gradually increased, reaching a maximum of 25 cells per mm2 on day 21, while a non-significant increase was observed at the subendocardial site. In the non-infarcted regions, the mast cell density increased transiently to reach a maximum of 7 cells per mm2 on day 35 in the subepicardial layer. Again, changes in mast cell density in the subendocardial layer were non-significant. In the sham group, a gradual increase to 9 cells per mm2 on day 21 and a subsequent decrease to 5 cells per mm2 on day 85 were observed in the subepicardial layers. These findings indicate a massive accumulation of mast cells in the subepicardial layers of the infarcted region and a small but significant effect of the surgical procedure on cardiac mast cell deposition, especially in the outer layers of the left ventricle.

Animals↗
PubMed1995

Purification and Characterization of Cystathionine (beta)-Lyase from Lactococcus lactis subsp. cremoris B78 and Its Possible Role in Flavor Development in Cheese.

An enzyme that degrades sulfur-containing amino acids was purified from Lactococcus lactis subsp. cremoris B78; this strain was isolated from a mixed-strain, mesophilic starter culture used for the production of Gouda cheese. The enzyme has features of a cystathionine (beta)-lyase (EC 4.4.1.8), a pyridoxal-5(prm1)-phosphate-dependent enzyme involved in the biosynthesis of methionine and catalyzing an (alpha),(beta)-elimination reaction. It is able to catalyze an (alpha),(gamma)-elimination reaction as well, which in the case of methionine, results in the production of methanethiol, a putative precursor of important flavor compounds in cheese. The native enzyme has a molecular mass of approximately 130 to 165 kDa and consists of four identical subunits of 35 to 40 kDa. The enzyme is relatively thermostable and has a pH optimum for activity around 8.0; it is still active under cheese-ripening conditions, viz., pH 5.2 to 5.4 and 4% (wt/vol) NaCl. A possible essential role of the enzyme in flavor development in cheese is suggested.

Journal Article↗