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W Engels

Publications and source records attributed to W Engels.

53 records · Page 3Linked to original sources

The nature of the competitive ability of spontaneous staphylocoagulase-negative mutants of Staphylococcus aureus with respect to growth of the parent strains in continuous culture.

During prolonged cultivation of S. aureus strains 104 and NCTC 8178 in continuous culture, staphylocoagulase-negative mutants arose and accumulated progressively in increasing proportions. The resulting loss of production of staphylocoagulase was accompanied by a simultaneous loss of production of alpha-haemolysin and PV-leucocidin. Characterization of the strains revealed no further difference in biotype, exoenzymes phage pattern and plasmid content. Cultivation in batch cultures showed that the maximal specific growth rates and specific oxygen-consumption rates of the mutant strains were slightly higher than those of the parent strains, whereas the production of total extracellular protein of the mutant strains had decreased significantly. From competition experiments between parent and mutant strains in chemostat cultures at different dilution rates and cultivation temperatures, it was concluded that the underlying mechanism of accumulation of staphylocoagulase-negative mutants in the chemostat is based on differences in affinity for the limiting substrate(s) rather than on differences in the production rates of total extracellular proteins. The complete repression of three exoenzymes, an partial repression of the total extracellular protein production, and an increased affinity for the limiting substrate(s) suggested that a mutation in a regulatory gene is involved. The possible role of a transposon in this mutation is discussed.

Culture Media↗

Secretion of staphylocoagulase be Staphylococcus aureus: the role of a cell-bound intermediate.

A cell-bound staphylocoagulase could be detected in chemostat cultures of Staphylococcus aureus 104 under magnesium-and oxygen-limited growth conditions. A distribution study revealed that 81% of the enzyme was membrane-bound and could be optimally released by Triton X-100. The remaining part was located in the periplasmic space and was released during protoplasting of organism. From inhibition studies with cerulenin, quinacrine, lincomycin an chloramphenicol, it was concluded that the cell-bound form was precursor in the secretion of extracellular staphylocoagulase. The involvement of a lipid intermediate/exoprotein-releasing protease system in the secretion of staphylocoagulase, and of exoproteins in general, is discussed.

Cell Membrane↗

Rapid and direct staphylocoagulase assay that uses a chromogenic substrate for identification of Staphylococcus aureus.

A rapid and direct assay of staphylocoagulase that uses chromogenic substrate Chromozym TH was developed for the routine identification of Staphylococcus aureus. False-positive and false-negative reactions were eliminated by measuring the activity of staphylocoagulase-prothrombin complex directly. The method was compared with three clotting assays. The results show that the new assay is specific, quantitative, and easily automated. If offers clinical laboratories a more reliable assay of staphylocoagulase and is a valuable aid for the rapid identification of S. aureus.

Blood Coagulation↗

Continuous culture studies on the production of staphylocoagulase by Staphylococcus aureus.

The production of staphylocoagulase was studied with continuous cultures of various S. aureus strains in a simple salts medium supplemented with mannitol, casein hydrolysate and three vitamins. Conditions of low oxygen availability and magnesium-limitation were required for optimal steady-state staphylocoagulase production. It was demonstrated that the specific rate of staphylocoagulase production was dependent on the growth rate. In two bovine strains, the production rate pattern was similar to that of an inducible enzyme sensitive to catabolite repression, although no specific inductor suppressor could be demonstrated. The human strain, on the other hand, produced staphylocoagulase constitutively. In all strains the specific rate of production of total extracellular protein was strictly proportional to the growth rate. The bovine strains produced 6 times more staphylocoagulase in chemostat culture as compared with batch cultures of the same organisms. It is likely that mannitol functioned as an energy source rather than as a carbon source because it was converted for a major part to acetate and for a minor part to lactate and not to new cell material. Repression and staphylocoagulase production by mannitol, acetate or lactate was not observed. The probable nature of the regulating mechanism(s) underlying staphylocoagulase production is discussed.

Animals↗

Influence of cultivation conditions on the production of staphylocoagulase by Staphylococcus aureus 104.

High yields of staphylocoagulase from Staphylococcus aureus 104 were obtained in a simple salts medium supplement with glycerol, casein hydrolysate and three vitamins. Conditions of oxygen-limitation (dissolved oxygen concentration less than 2%), a pH of 7.4, a temperature of 35 degrees C and a 1 in 10 inoculum of overnight culture were required for optimal yields of staphylocoagulase.

Caseins↗

Occurrence of intercellular bridges between follicle epithelial cells in the ovary of Apis mellifica queens.

Hitherto unknown intercellular bridges or fusomes between the follicle epithelial cells investing the oocytes of Apis mellifica queens have been observed both with light and electron microscopy. Usually each follicle cell has 2-3 intercellular bridges. In surfacial paraffin sections, the intercellular bridges can be seen to connect a series of follicle cells which may be branching. The intercellular bridges lie close to the egg cortex and this position is relatively constant. The width of the fusomal ring canal varies in different developmental stages. In stages 3 and 4 of oogenesis, which are the main vitellogenic stages, the intercellular bridges measure 0-5 micron, while in stages 1 and 2 they have a diameter ranging from 1-5 to 3-5 micron. In these stages the intercellular bridges are provided with numerous transverse microfilaments which disappear later. The fusomal lips are thickened and consist of electron-dense material and an additional layer of less electron-dense material both inside and outside. Ribosomes flow across the bridge. The intercellular bridges may serve to synchronize the differentiation and functional activity of the follicle epithelium during the course of oogenesis.

Animals↗

Significance of myocardial eicosanoid production.

The precise role of eicosanoids in the development of myocardial injury during ischemia and reperfusion is still a matter of debate. Enhanced local production of these bioactive compounds appears to be a common response to tissue injury. Most likely, the cardiac tissue has the capacity to generate prostaglandins, thromboxanes as well as leukotrienes. Prostacyclin (PGI2) is the major eicosanoid produced by the jeopardized myocardium. In addition, at sites of tissue injury activation of platelets and infiltrating leukocytes results in the formation of considerable amounts of thromboxanes and leukotrienes. The production of eicosanoids requires prior release of arachidonic acid (AA) from phospholipids. Both ischemia and reperfusion are associated with a rise in the tissue level of AA. The absence of a proportional relationship between the tissue level of AA and the amounts of PGI2 produced suggests that the sites of AA accumulation and PGI2 formation are different. It is conceivable that AA accumulation is mainly confined to myocytes, whereas the capacity to synthesize PGI2 mainly resides in vascular cells. Both beneficial and detrimental effects of eicosanoids on cardiac tissue have been described. Prostaglandins act as vasodilators. Besides, some of the prostaglandins, especially PGI2, are thought to possess cyto-protective properties. Thromboxanes and leukotrienes may impede blood supply by increasing smooth muscle tone. Besides, leukotrienes augment vascular permeability. Experimental studies, designed to evaluate the effect of pharmacological agents, like PGI2-analogues and lipoxygenase and cyclo-oxygenase inhibitors, indicate that eicosanoids influence the outcome of myocardial injury.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗