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This paper describes the principle of a homogeneous indirect fluorescence quenching immunoassay that uses monoclonal antibodies. It is a carrier-free assay system that is performed completely in solution. The assay system was established for the determination of a low molecular weight substance (hapten), the herbicide diuron, used as a model analyte. A fluorescein-monuron conjugate together with a fluorescence-quenching monoclonal anti-fluorescein antibody and an anti-analyte antibody (here an anti-diuron/monuron monoclonal antibody) were used as central components of the assay. The fluorescein-monuron conjugate can be bound either by the anti-fluorescein monoclonal antibody or by the anti-diuron/ monuron monoclonal antibody. Due to steric hindrance, binding of both antibodies to the conjugate was not possible at the same time. By selecting the antibody concentrations appropriately, a dynamic equilibrium can be established that permits the preferential binding of the anti-diuron/monuron antibody to the conjugate, which allows the fluorescein in the conjugate to fluoresce. This equilibrium can be easily altered by adding free analyte (diuron), which competes with the conjugate to bind to the anti-diuron/monuron antibody. A reduction of anti-diuron/monuron antibody binding to the conjugate results in an increase in the binding of the anti-fluorescein antibody, which leads to a decrease in the fluorescence of the conjugate. The fluorescence is therefore a direct indicator of the state of equilibrium of the system and thus also the presence of free unconjugated analyte. The determination of an analyte based on this test principle does not require any washing steps. After the test components are mixed, the dynamic equilibrium is rapidly reached and the results can be obtained in less than 5 min by measuring the fluorescence of the fluorescein. We used this test principle for the determination of diuron, which was demonstrated for concentrations of approximately 5 nM.
KEPI is a protein kinase C-potentiated inhibitory protein for type 1 Ser/Thr protein phosphatases. We found no or reduced expression of KEPI in breast cancer cell lines, breast tumors and metastases in comparison to normal breast cell lines and tissues, respectively. KEPI protein expression and ubiquitous localization was detected with a newly generated antibody. Ectopic KEPI expression in MCF7 breast cancer cells induced differential expression of 95 genes, including the up-regulation of the tumor suppressors EGR1 (early growth response 1) and PTEN (phosphatase and tensin homolog), which is regulated by EGR1. We further show that the up-regulation of EGR1 in MCF7/KEPI cells is mediated by MEK-ERK signaling. The inhibition of this pathway by the MEK inhibitor UO126 led to a strong decrease in EGR1 expression in MCF7/KEPI cells. These results reveal a novel role for KEPI in the regulation of the tumor suppressor gene EGR1 via activation of the MEK-ERK MAPK pathway.
Active host-searching in bat ectoparasites. As a pilot study, this paper discribes the mobility of ectoparasites after removing from their hosts. Spinturnix acuminatus (Koch, 1836)(Acari, Spinturnicidae) a permanent stationary bat mite normally survives the removing only a few hours. This species moves easily on the patagium or on human skin. In contrast, on other surfaces the movements of this species seems to be less orientated and it is unlikely that the mite can reach a host in some distance. Nycteribia kolenatii Theodor & Moscona, 1954 (Diptera: Nycteribiidae) a bat fly, shows excellent movements in the fur of its host. The specimens survive the removing from the host several days. The ability to clamber up a wall was tested with a rough clay brick. The bat fly achieves only 9,6 cm on average. This result does not support good abilities for host searching in this species. Bat fleas (Siphonaptera: Ischnopsyllidae): Ischnopsyllus elongatus (Curtis, 1832), I. hexactenus (Kolenati, 1856), I. simplex (Rothschild, 1906)and other species survive the removing from their hosts several days. In case of a vertical barrier they showed a stereotype upside climbing, followed by a horizontal appetitive behavior. The fleas were climbing a distance in the upside of 6,45 m on average(1,2-21,68 m). As excellent climbers bat fleas are able to overcome edges and slopes and to crawl on the ceiling. Therefore, it seems to be no problem for a flea to parasite any bat in a resting room. Additionally, some bugs (Cimex lectularius Linnaeus, 1758) were examinated. They run with a speed of 63 cm per minute and were able to climb easily on the brick. On average their stretch of way achieved 2,85 m in ten minutes.
Die vorliegende Untersuchung von Fledermaeusen in Sommer- und Winterquartieren liefert neue daten zur Verbreitung und Haeufigkeit von Ektoparasiten aus Brandenburg und anderen Bundeslaendern. An 11 Fledermausarten wurden insgesamt 7 Floharten (Ischnosyllidae), zwei Lausfliegenarten (Nycteribiidae) und zwei Wanzenarten (Cimicidae) nachgewiesen. Die Floharten Ischnopsyllus intermedius und die Artengruppe I. simplex/mysticus ergaenzen die Liste der aktuellen Funde fuer Brandenburg. Nach dem milden Winter 2006-7 wurden in einigen Winterquartieren Veraenderungen im Artenspektrum der Parasiten festgestellt. Nycteribia kolenatii an Myotis daubentoni und Nycteribia eusarca an Nyctalus noctula traten bei der vorliegenden Untersuchung relativ haeufig auf. Die anderen ektoparasitischen Floehe und Lausfliegen waren sehr selten zu finden. Die meisten Parasitenarten und die hoechste Befallsintensitaet wies die Wasserfledermaus (Myotis daubentoni) auf.
Annual plants under cyclic disturbance regime : better understanding through model aggregation
(2008)
In their application for conservation ecology, 'classical' analytical models and individual-based simulation models (IBMs) both entail their specific strengths and weaknesses, either in providing a detailed and realistic representation of processes or in regard to a comprehensive model analysis. This well-known dilemma may be resolved by the combination of both approaches when tackling certain problems of conservation ecology. Following this idea, we present the complementary use of both an IBM and a matrix population model in a case study on grassland conservation management. First, we develop a spatially explicit IBM to simulate the long-term response of the annual plant Thlaspi perfoliatum (Brassicaceae), claspleaf pennycress, to different management schemes (annual mowing vs. infrequent rototilling) based on field experiments. In order to complement the simulation results by further analyses, we aggregate the IBM to a spatially nonexplicit deterministic matrix population model. Within the periodic environment created by management regimes, population dynamics are described by periodic products of annual transition matrices. Such periodic matrix products provide a very conclusive framework to study the responses of species to different management return intervals. Thus, using tools of matrix model analysis (e.g., loop analysis), we can both identify dormancy within the age-structured seed bank as the pivotal strategy for persistence under cyclic disturbance regimes and reveal crucial thresholds in some less certain parameters. Results of matrix model analyses are therefore successfully tested by comparing their results to the respective IBM simulations. Their implications for an enhanced scientific basis for management decisions are discussed as well as some general benefits and limitations of the use of aggregating modeling approaches in conservation.
Growing recognition of the importance of long-distance dispersal (LDD) of plant seeds for various ecological and evolutionary processes has led to an upsurge of research into the mechanisms underlying LDD. We summarize these findings by formulating six generalizations stating that LDD is generally more common in open terrestrial landscapes, and is typically driven by large and migratory animals, extreme meteorological phenomena, ocean currents and human transportation, each transporting a variety of seed morphologies. LDD is often associated with unusual behavior of the standard vector inferred from plant dispersal morphology, or mediated by nonstandard vectors. To advance our understanding of LDD, we advocate a vector-based research approach that identifies the significant LDD vectors and quantifies how environmental conditions modify their actions.
Sf6 belongs to the Podoviridae family of temperate bacteriophages that infect gram-negative bacteria by insertion of their double-stranded DNA. They attach to their hosts specifically via their tailspike proteins. The 1.25 Å crystal structure of Shigella phage Sf6 tailspike protein (Sf6 TSP) reveals a conserved architecture with a central, right-handed ; helix. In the trimer of Sf6 TSP, the parallel ; helices form a left-handed, coiled;; coil with a pitch of 340 Å. The C-terminal domain consists of a ; sandwich reminiscent of viral capsid proteins. Further crystallographic and biochemical analyses show a Shigella cell wall O-antigen fragment to bind to an endorhamnosidase active site located between two ;-helix subunits each anchoring one catalytic carboxylate. The functionally and structurally related bacteriophage, P22 TSP, lacks sequence identity with Sf6 TSP and has its active sites on single subunits. Sf6 TSP may serve as an example for the evolution of different host specificities on a similar general architecture.