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Biomedical subjects

B Streit

Publications and source records attributed to B Streit.

At least 19 recordsLinked to original sources

Knipholone, a selective inhibitor of leukotriene metabolism.

Inhibition of leukotriene formation is one of the approaches to the treatment of asthma and other inflammatory diseases. We have investigated knipholone, isolated from the roots of Kniphofia foliosa, Hochst (Asphodelaceae), for inhibition of leukotriene biosynthesis in an ex vivo bioassay using activated human neutrophile granulocytes. Moreover, activities on 12-lipoxygenase from human platelets and cycloxygenase (COX)-1 and -2 from sheep cotyledons and seminal vesicles, respectively, have been evaluated. Knipholone was found to be a selective inhibitor of leukotriene metabolism in a human blood assay with an IC(50) value of 4.2microM. However, at a concentration of 10microg/ml, the compound showed weak inhibition of 12(S)-HETE production in human platelets and at a concentration of 50microM it produced no inhibition of COX-1 and -2. In our attempt to explain the mechanism of inhibition, we examined the antioxidant activity of knipholone using various in vitro assay systems including free radical scavenging, non-enzymatic lipid peroxidation, and metal chelation. Knipholone was found to be a weak dose-independent free radical scavenger and lipid peroxidation inhibitor, but not a metal chelator. Therefore, the leukotriene biosynthesis inhibitory effect of knipholone was evident by its ability either to inhibit the 5-lipoxygenase activating protein (FLAP) or as a competitive (non-redox) inhibitor of the enzyme. Cytotoxicity results also provided evidence that knipholone exhibits less toxicity for a mammalian host cell.

Animals↗

Eel parasite diversity and intermediate host abundance in the River Rhine, Germany.

European eels (Anguilla anguilla) from 2 sampling sites on the Rhine river (near Karlsruhe and near Worms) were investigated with respect to their parasite communities. Nine different metazoan species were found to live in and on the eels. The highest number of species was recorded from the intestine, which contained up to 6 different helminths. Among these, acanthocephalans were the most prevalent worms with the eel-specific parasite Paratenuisentis ambiguus as the dominant species of the intestinal component communities at both sites. Comparing the intestinal parasites from eels caught near Karlsruhe with those from Worms, the acanthocephalans showed a significantly lower abundance at Worms. A significantly lower mean number of intestinal helminth species as well as a significantly lower Brillouin's Index was found at Worms compared with Karlsruhe. This difference could be related to the abundance of the respective intermediate crustacean hosts. At the sampling site Worms the amphipod Corophium curvispinum was the dominant crustacean. Additionally, only the isopod Jaera istri and the amphipod Dikerogammarus villosus were found. All these crustacean species have only recently colonized the Rhine river system via the Main-Danube canal, built in the early 1990s. They are not known to act as intermediate hosts for any of the acanthocephalans found in the eels. The site near Karlsruhe exhibited a higher crustacean diversity, including Asellus aquaticus and different species of the genus Gammarus, which are all known intermediate hosts for the acanthocephalans found. Therefore, changes of eel parasite diversity can be correlated with the appearance of invading crustacean species (neozoans).

Anguilla↗

Isolation of Hox genes from the scyphozoan Cassiopeia xamachana: implications for the early evolution of Hox genes.

The isolation of Hox genes from two cnidarian groups, the Hydrozoa and Anthozoa, has sparked hypotheses on the early evolution of Hox genes and a conserved role for these genes for defining a main body axis in all metazoan animals. We have isolated the first five Hox genes, Scox-1 to Scox-5, from the third cnidarian class, the Scyphozoa. For all but one gene, we report full-length homeobox plus flanking sequences. Four of the five genes show close relationship to previously reported Cnox-1 genes from Hydrozoa and Anthozoa. One gene, Scox-2, is an unambiguous homologue of Cnox-2 genes known from Hydrozoa, Anthozoa, and also Placozoa. Based on sequence similarity and phylogenetic analyses of the homeobox and homeodomain sequences of known Hox genes from cnidarians, we suggest the presence of at least five distinct Hox gene families in this phylum, and conclude that the last common ancestor of the Recent cnidarian classes likely possessed a set of Hox genes representing three different families, the Cnox-1, Cnox-2, and Cnox-5 families. The data presented are consistent with the idea that multiple duplication events of genes have occurred within one family at the expense of conservation of the original set of genes, which represent the three ancestral Hox gene families.

Amino Acid Sequence↗

Short-term toxicity of lindane, hexachlorobenzene, and copper sulfate to tubificid sludgeworms (Oligochaeta) in artificial media.

The toxicity of lindane, hexachlorobenzene, and copper sulfate to Tubifex tubifex and Limnodrilus hoffmeisteri was determined using an easily applicable and standardizable 72-h short-term test system. It was designed for the quick assessment of sublethal and lethal effects of sediment-associated chemicals on the worms. An artificial sediment based on the Artificial Soil according to OECD Guideline No. 207 was used as test medium. The data confirm the common view that oligochaetes are highly tolerant of lethal effects. However, sublethal effects were detected at considerably lower concentrations than found for lethal effects. The EC50 values for autotomy (172 mg/kg dry wt sediment) and sediment avoidance (217 mg/kg) for T. tubifex exposed to lindane-contaminated sediment were, for example, more than five times lower than the LC50 value (> 1000 mg/kg). The no-observed-effect concentration for reworking activity (8 mg/kg) was more than 125 times lower than the LC50. Tubificids thus turned out to represent useful test organisms for the assessment of the ecotoxicological hazard potential of chemicals in the sediment compartment, because the sublethal effects not only affect the individual, but can influence the population levels and, consequently, the composition of the benthic community.

Animals↗

Bioaccumulation of contaminants in fish.

The term bioaccumulation is defined as uptake, storage, and accumulation of organic and inorganic contaminants by organisms from their environment. Bioaccumulation therefore results from complex interactions between various routes of uptake, excretion, passive release, and metabolization. For fish, the bioaccumulation process includes two routes of uptake: aqueous uptake of water-borne chemicals, and dietary uptake by ingestion of contaminated food particles. The contribution to bioaccumulation that results from aqueous exposure and is taken up by the gills is called bioconcentration. The contribution to bioaccumulation resulting from dietary exposure via uptake by intestinal mucosa is termed biomagnification. In both cases, important co-determinants for bioaccumulation are the various elimination mechanisms. This chapter presents a short historical survey of the problem of bioaccumulation with particular reference to fish and of the various approaches to study bioaccumulation. This is followed by an overview of our present knowledge about basic physico-chemical determinants that either increase or reduce the bioaccumulation potential of various chemicals, and about the physiological basis of gills, blood circulation and intestines, as far as they are crucial for our understanding of uptake and accumulation. Finally, selected quantitative data and modelings of bioaccumulation in fish will be discussed, with regard to such problems as the relative importance of aqueous and dietary uptake.

Animals↗

Induction of rapid IL-1 beta mRNA degradation in THP-1 cells mediated through the AU-rich region in the 3'UTR by a radicicol analogue.

A radicicol analogue (analogue A) was found to inhibit interleukin 1 beta (IL-1 beta) and tumour necrosis factor alpha (TNF-alpha) secretion from THP-1 cells. If added to cells activated by interferon gamma and lipopolysaccharide, radicicol analogue A not only inhibited the secretion of IL-1 beta but also induced an extremely rapid degradation of IL-1 beta, IL-6 and TNF-alpha mRNA to undetectable levels within 5-8 h. This degradation is independent of translation and of the signal inducing transcription. The common feature of these genes is the inclusion of one or more copies of the mRNA-instability sequence, AUUUA, in the 3' untranslated region. Indeed, no destabilizing effect of radicicol analogue A could be observed on mRNA derived from the expression of an IL-1 beta construct lacking the AUUUA motifs of the 3'UTR. The effect of radicicol analogue A on protein/mRNA interaction and on post-translational modifications of cytoplasmic proteins is described. This class of compound constitutes a valuable tool for the further elucidation of the mechanism of mRNA degradation of cytokines and proto-oncogenes.

Base Sequence↗

Homeobox genes in the cnidarian Eleutheria dichotoma: evolutionary implications for the origin of Antennapedia-class (HOM/Hox) genes.

In order to test current hypotheses on the early evolution of Antp-class (HOM/Hox) genes in metazoan animals, we surveyed the diploblastic hydrozoan Eleutheria dichotoma for the presence of these genes. By means of PCR with different sets of degenerate primers, three new homeobox sequences were identified, in addition to two previously found fragments. Following RACE amplification of the 3' and 5' flanking regions, we classified all five genes, Cnox-1 to Cnox-5, as belonging to the Antp-class. Homeodomain identity to the most similar HOM/Hox genes from Drosophila and human ranges from 58 to 72%. Each of the Cnox genes contains diagnostic residues of more than one Antp-class family known from triploblastic animals. These data are at odds with a previously proposed ancestral HOM/Hox cluster composition, with respect to both the number and types of Antp-class homeobox genes found.

Amino Acid Sequence↗

The effects of nested primer binding sites on the reproducibility of PCR: mathematical modeling and computer simulation studies.

The polymerase chain reaction (PCR) has become an indispensable tool in modern biological research. Although the application of PCR is a standard routine, we widely lack a theoretical understanding of the dynamic processes involved, especially with respect to the amplification of nonreproducible and/or unexpected amplification products. For one potential source of uncertainty, the presence of nested primer binding sites within an amplifyable DNA locus, we consider a simple stochastic model for the dynamics of PCR amplification of competing products. For commonly used thermostable DNA polymerases lacking a 5'-3'-exonuclease activity, we predict the relative amplification frequencies of competing PCR products dependent on the primer binding probability, the number of PCR cycles, and the number of initial DNA template molecules. At low primer binding probabilities and low numbers of initial DNA template molecules and PCR cycles, we expect the amplification of two products. At high primer binding probabilities and/or high copy numbers of initial template molecules only one main amplification product is predicted at increasing cycle numbers. Furthermore, by means of computer simulation studies we quantify the stochastic variation for the amplification frequencies of competing products.

Bacteriophage lambda↗

RAPD identification of microsatellites in Daphnia.

Simple sequence repeats (SSRs, or microsatellites) have been constantly gaining importance as single-locus DNA markers in population genetics and behavioural ecology. We tested a PCR-based strategy for finding microsatellite loci in anonymous genomes, which avoids genomic library construction and screening, and the need for larger amounts of DNA. In the first step, parts of a genome are randomly amplified with arbitrary 10mer primers using RAPD fingerprinting. Labelled SSR-oligonucleotides serve as probes to detect complementary sequences in RAPD products by means of Southern analyses. Subsequently, positive RAPD fragments of suitable size are cloned and sequenced. Using GA and GT probes, we applied this approach to waterfleas (Daphnia) and revealed 37 hybridization signals in 20 RAPD profiles. Thirteen positive RAPD fragments from three Daphnia species and two hybrid 'species' were cloned and sequenced. In all cases simple sequence repeats were detected. We characterized seven perfect repeat loci, which were found to be polymorphic within and between species.

Animals↗

Outcrossing rates and correlated matings in a predominantly selfing freshwater snail.

We estimated outcrossing rates and correlated matings in a natural population of the hermaphroditic freshwater snail Ancylus fluviatilis, using progeny arrays and polymorphic allozyme markers. This design circumvents drawbacks of earlier studies, and yields maximum likelihood estimates of outcrossing rates at both the individual and population level. Adult snails were sampled during the spring breeding period, allowing for prior copulations and sperm transfer under natural conditions. Once in the laboratory, they were kept isolated for 21 days and laid egg capsules, and emerging hatchlings from 42 families were scored for five polymorphic loci. 87 of 848 offspring exhibited non-maternal alleles, representing a minimum population-level outcrossing of 10.3%. Maximum likelihood estimates are in the range 13-15%. Importantly, there is significant among-family heterogeneity, as only 16 of 42 families are estimated to have outcrossed, in accord with high estimates of correlated selfing within progeny arrays. Moreover, the high proportion of full sibs among outcrossed sibs suggests a limited extent of multiple paternity. Our data reveal important heterogeneity in the mating system of an animal hermaphrodite, and indicate the potential for evolutionary change in the breeding system.

Animals↗

The evolutionary ecology of Daphnia.

In order to generate genetic markers from both nuclear and mitochondrial DNA, we used three PCR-based techniques (RAPD, mtDNA-RFLPs, and sequencing of an amplified mtDNA fragment) to illuminate various aspects of the population genetics of large-lake Daphnia species. Estimations of genetic diversity at different taxonomic levels integrated with ecological data revealed insights into the genetic components of the evolutionary process of interspecific hybridization in these Daphnia species, which had previously been documented with allozyme markers. Our new molecular data suggest the occurrence of recent hybridization and backcrossing events, and allow the identification of the maternal species of hybrid clones.

Animals↗

Natural hybridization in freshwater animals. Ecological implications and molecular approaches.

The number of cases where the phenomena of hybridization and gene introgression have been found in species interactions is steadily increasing, in both plant and animal taxa. During the last few years, many examples have been detected even in otherwise well-known freshwater animal taxa. We discuss the topic with respect to ecology and evolutionary processes and compare the main potentials and limitations of allozymes, mitochondrial DNA, and RAPD markers to address some important genetic issues of interspecific hybridization in natural populations of selected freshwater model systems.

Amphibians↗

Bioaccumulation processes in ecosystems.

The fate of environmental pollutants--the various isotopes of elements, and inorganic or organic compounds--is a fundamental aspect of ecology and ecotoxicology, and bioaccumulation is a phenomenon often discussed in this context. Human activities have drastically altered natural concentrations of many substances in the environment and added numerous new chemicals. An understanding of the processes of bioaccumulation is important for several reasons. 1) Bioaccumulation in organisms may enhance the persistence of industrial chemicals in the ecosystem as a whole, since they can be fixed in the tissues of organisms. 2) Stored chemicals are not exposed to direct physical, chemical, or biochemical degradation. 3) Stored chemicals can directly affect an individual's health. 4) Predators of those organisms that have bioaccumulated harmful substances may be endangered by food chain effects. While former theories on the processes of bioaccumulation focused on single aspects that affect the extent of accumulation (such as the trophic level within the food chain or the lipophilicity of the chemical), modern theories are based on compartmental kinetics and the integration of various environmental interactions. Concepts include results from quantitative structure-activity relationships (QSAR), pharmacokinetics, ecophysiology and general biology, molecular genetic aspects and selection, and finally the structure of communities and man-made alterations in them.

Animal Population Groups↗

Establishment of an enzyme-linked immuno-sorbent assay for urinary trypsin inhibitor by using a monoclonal antibody.

Monoclonal antibodies against inter-alpha-trypsin-inhibitor (ITI) were produced. One clone showing specificity for urinary trypsin inhibitor (UTI), a proteolytic fragment of ITI, which is excreted into urine, was selected for the establishment of an enzyme-linked immuno-sorbent assay (ELISA). The ELISA for the quantification of UTI was shown to work reproducibly in the range between 0.5 and 10 ng UTI/ml urine. Urines of several patients suffering from different lung diseases were screened for UTI using the established ELISA. Highest UTI levels were found in the urine of patients with lung empyema. A more moderate increase was observed in patients suffering from lung tuberculosis and from secondary and primary lung tumors.

Alpha-Globulins↗

Transurethral resection of the carcinomatous ureteral orifice (Analysis of 27 operations).

Transurethral electroresection for malignant involvement of the ureteral orifice was carried in 17 cases. In the light of published data the results may be regarded as favourable. It is stated that in case of operability resection of the orifice represents a radical procedure, superior to open techniques, and it has no adverse consequences in the majority of cases.

Adult↗