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Biomedical subjects

C Schroeder

Publications and source records attributed to C Schroeder.

105 records · Page 6Linked to original sources

The inhibitory effect of aspirin on human endothelial cells.

Human endothelial cell monolayers prepared from umbilical veins have been incubated with aspirin (1--2 mM) dissolved in Hepes modified solution and in platelet-rich plasma. They have also been incubated with plasma prepared from subjects before and after intake of aspirin giving a mean plasma concentration of 0.5 mM. The effects of the endothelial cells on ADP and collagen-induced platelet aggregation and malondialdehyde production in platelet-rich plasma have been tested. The endothelial cells had a spontaneous inhibitory effect on all three parameters. This effect was abolished when the cells were incubated with aspirin dissolved in MHS for 20 min and the increase in effect observed when platelet-rich plasma was incubated with endothelial cells for a period of 30 min was similarly inhibited when aspirin was dissolved in plasma or when plasma prepared from subjects who had taken aspirin were used. Aspirin had no inhibitory effect on prostacyclin (PGI2) with regard to the effect of PGI2 on platelets. On the contrary, the two compounds had an additive inhibitory effect on platelet aggregation induced by ADP and collagen. These findings should be considered with regard to the use of aspirin as an antithrombotic agent.

Aspirin↗

Active protection by bacteriophages T3 and T7 against E. coli B- and K-specific restriction of their DNA.

The bacteriophages T3 and T7 are not modified and restricted by E. coli strains with different host specificity (E. coli B, K, O) in vivo. The phages code for a gene product with the ability to overcome classical restriction (ocr): ocr- mutants are subject to modification and restriction via DNA methylation vs cleavage. The T3 genome possesses recognition sites for the restriction endonuclease R.EcoB which, unless the DNA is B-specifically modified, trigger 5-7 DNA cleavages. The ocr gene function of T3 and T7 is located within the gene 0.3 region of these phages and is not identical with the sam (SAMase) function of T3. The mechanism of ocr protection remains unclear, while it is certain that this protection by the gene 0.3 protein is exerted in the infected cell and not through "over-all" modification in the preceding growth cycle of the phage.

Adenosylmethionine Decarboxylase↗

Host-dependent modification of bacteriophage T7 and SAMase-negative T3 derivatives affecting their adsorption ability.

When passaging phage T7 and SAMase-negative T3 mutants between E. coli strains with identical (EcoB) or without (EcoO) DNA host specificity, phenotypically a host-controlled modification and restriction is observed. This phenomenon is not due to "classical" modification and restriction of the bacteriophage DNA but depends on the reversibly altered adsorption capacity of the phages on the different host strains.

Adenosylmethionine Decarboxylase↗

Effects of 3'-fluorothymidine, nalidixic acid, and mitomycin C on temperature-sensitive mutants of the DNA polymerase of phage T4.

The temperature-sensitive DNA polymerase mutator mutants A58 and L98 are less inhibited by 3'-Fluorothymidine than the T4 wild type and the antimutator mutant CB121, in consequence of the assumed differences between the polymerase-associated exonuclease activities. This interpretation is confirmed by results with nalidixic acid and mitomycin C. The use of systems of different temperature-sensitive mutants of one or more genes is proposed for 1.) investigating the mode of action of drugs, 2.) studies on the mechanism of enzyme action and the functions affected in temperature-sensitive enzymes, and 3.) for enzyme-specific drug screening.

Chloramphenicol↗

Avoidance of DNA methylation. A virus-encoded methylase inhibitor and evidence for counterselection of methylase recognition sites in viral genomes.

The ocr+ gene of bacterial virus T7 codes for the first protein recognized to inhibit a specific group of DNA methylases. The recognition sequences of several other DNA methylases, not susceptible to Ocr inhibition, are significantly suppressed in the virus genome. The bacterial virus T3 encodes an Ado-Met hydrolase, destroying the methyl donor and causing T3 DNA to be totally unmethylated. These observations could stimulate analogous investigations into the regulation of DNA methylation patterns of eukaryotic viruses and cells. For instance, an underrepresentation of methylation sites (5'-CG) is also true for animal DNA viruses. Moreover, we were able to disclose some novel properties of DNA restriction-modification enzymes concerning the protection of DNA recognition sequences in which only one strand can be methylated (e.g., type III enzyme EcoP15) and the primary resistance of (unmethylated) DNA recognition sites towards type II restriction endonuclease EcoRII.

Base Sequence↗

Hyperacute rejection is attenuated in GalT knockout swine lungs perfused ex vivo with human blood.

BACKGROUND: Hyperacute rejection (HAR) is one of the principal obstacles to successful xenotransplantation. Homozygous alpha-1,3-galactosyltransferase knockout (GalT-KO) miniature swine now offer the prospect of overcoming this barrier to xenotransplantation. In this study, the short-term function of GalT-KO swine lungs was evaluated in a well-established ex vivo model of swine-to-human lung xenotransplantation. METHODS: Lungs from homozygous GalT-KO swine (n = 3) and control lungs from pigs of the background strain used to create the GalT-KO pig line (n = 2) were perfused ex vivo with freshly collected heparinized human blood. Graft function was assessed by various physiologic measurements, serial histologic and immunohistochemical evaluation, and assays of complement and platelet activation. RESULTS: Xenoperfused control swine lungs exhibited HAR with graft survival times <5 minutes. In contrast, GalT-KO swine lungs retained their function for approximately 2 hours, on average. GalT-KO swine lungs showed decreased complement and platelet activation compared with controls. Nonetheless, activation of complement and coagulation cascades was not completely eliminated in the GalT-KO swine lungs. CONCLUSIONS: The survival of xenoperfused GalT-KO swine lungs was significantly prolonged, as compared with control lungs expressing Gal. This appears to have been due largely to substantially reduced complement activation. Nonetheless, the xenoperfused GalT-KO lungs still showed some evidence of complement fixation and intravascular coagulopathy by the time of graft demise.

Acute Disease↗

Evaluation of methodological and biological influences on the collection and composition of exhaled breath condensate.

The purpose of this inter-species comparison (calves and pigs) was to identify methodological and biological influences on the collection and composition of exhaled breath condensate (EBC). A total of 352 EBC samples were collected, whilst variables of ventilation were registered in parallel. Partial pressure of carbon dioxide (pCO2) and pH were analysed in non-degassed EBC samples. The concentration of total protein in EBC was measured colorimetrically. In both species, lung function was evaluated before and after EBC collection. Statistical analyses were performed to study the effect of EBC collection on lung function and to identify the influence of ventilatory variables on the collection and composition of EBC. Collection of EBC did not affect lung function. Despite the volume of EBC collected per unit time being primarily dependent on ventilation per unit time, species-specific conditions during the EBC collection process resulted in different dependences of EBC collection from other variables of ventilation (i.e. maximal airflow during expiration or expired tidal volume kg-1 body weight). The concentration of protein ml(-1) EBC increased with the expired volume per min and with peak expiratory flow. Although the pCO2 in fresh EBC was significantly negatively dependent on the duration of collection, comparable pHs (5.6 - 6.2) were measured in EBC of both calves and pigs. The obtained data may help one standardize EBC collection in different species.

Animals↗

Enhanced basal and stimulated PMN chemiluminescence activity in children with atopic dermatitis: stimulatory role of colonizing staphylococci?

In adults, intense staphylococcal skin colonization and hyperactivity of polymorphonuclear leukocyte (PMN) oxidative metabolism are characteristic features of atopic dermatitis. Precise data on childhood atopic dermatitis are lacking. In a prospective study we analysed the PMN chemiluminescence activity with special reference to staphylococcal stimuli in 19 children (mean age 6.2 years) with mild to moderate atopic dermatitis. Staphylococcus aureus was isolated from 17/19 (90%) of children with atopic dermatitis and 13/45 (29%) of healthy age-matched controls (p less than 0.001). The mean (SEM) chemiluminescence activity of unstimulated atopic dermatitis-PMN was 0.34 (0.009) (controls: 0.092 (0.003) x 10(6) cpm/10(6) PMN/min (p less than 0.02). Staphylococcal antigens (S. aureus, S. epidermidis, S. haemolyticus) induced a 1.9-3.1-fold higher peak chemiluminescence response in children with atopic dermatitis than in controls (p less than 0.05). The time interval until peak chemiluminescence activity was considerably shorter for all stimuli in atopic dermatitis. We conclude that PMN of children with atopic dermatitis are "primed", showing enhanced release of reactive oxygen metabolites even in the "resting" state, and are easily stimulated by staphylococcal antigens present on the skin of patients with atopic dermatitis from early childhood on. We speculate that PMN hyperreactivity may contribute to chronic skin damage in atopic dermatitis.

Adolescent↗

Mutual activation of two restriction endonucleases: interaction of EcoP1 and EcoP15.

Type III restriction endonucleases recognize nonsymmetric nucleotide sequences. A necessary condition for DNA cleavage is the presence of two unmethylated recognition sites which are inversely ('head-to-head') oriented in the DNA double strand. A DNA substrate possessing one EcoP1 and one EcoP15 site in the head-to-head configuration could not be cleaved by the individual enzymes, however, it was specifically digested in the simultaneous presence of both enzymes. In agreement with the tracking-collision model for the DNA interaction of type III enzymes cleavage could be abolished by Lac repressor bound between the two sites. We conclude that two different type III enzymes can functionally cooperate in the cleavage of DNA.

DNA Methylation↗