Chromosome Evolution in Bats as Revealed by FISH: The Ongoing Search for the Ancestral Chiropteran Karyotype

Chiroptera, the second largest order of mammals, comprises more than 1,000 species in 18 highly morphologically diverse families. Chromosome painting with human probes has been applied to 10 bat species from 8 families. Except for the combination 10/12pq/22q, all syntenic segmental associations proposed for the mammalian ancestor have been found in Chiroptera. Bat-specific painting probes, established from 4 species of 3 families, have been used in whole chromosome painting experiments in 29 species from 8 families. The results show that the prevailing mode of chromosomal evolution in bats is Robertsonian translocation with a large number of convergent events. Given our present knowledge of chiropteran karyotypes, only a few elements of the ancestral chiropteran karyotype can be reconstructed with confidence.     

Binding of complement inhibitor C4b-binding protein contributes to serum resistance of Porphyromonas gingivalis

The periodontal pathogen Porphyromonas gingivalis is highly resistant to the bactericidal activity of human complement, which is present in the gingival crevicular fluid at 70% of serum concentration. All thirteen clinical and laboratory P. gingivalis strains tested were able to capture the human complement inhibitor C4b-binding protein (C4BP), which may contribute to their serum resistance. Accordingly, in serum deficient of C4BP, it was found that significantly more terminal complement component C9 was deposited on P. gingivalis. Moreover, using purified proteins and various isogenic mutants, we found that the cysteine protease high molecular weight arginine-gingipain A (HRgpA) is a crucial C4BP ligand on the bacterial surface. Binding of C4BP to P. gingivalis appears to be localized to two binding sites: on the complement control protein 1 domain and complement control protein 6 and 7 domains of the alpha-chains. Furthermore, the bacterial binding of C4BP was found to increase with time of culture and a particularly strong binding was observed for large aggregates of bacteria that formed during culture on solid blood agar medium. Taken together, gingipains appear to be a very significant virulence factor not only destroying complement due to proteolytic degradation as we have shown previously, but was also inhibiting complement activation due to their ability to bind the complement inhibitor C4BP.     

A Site-Specific Approach for the Evaluation of Natural Attenuation at Metals-Impacted Sites

Consideration of monitored natural attenuation (MNA) as a remedy component for metals-contaminated sites can be achieved using a site-specific screening approach, followed by application of one or a series of sequential extraction measurements. Hazardous waste sites contaminated with metals can be screened for the implementation of monitored natural attenuation on the basis of contaminant-specific soil chemical characteristics (i.e., K_d's, solubilities, and nonexchangeable sorbed fraction). Field cases are used to demonstrate the screening approach and to outline the primary considerations involved in accurately applying sequential extraction procedures to support the of MNA for site remediation. The results of these case studies provide strong evidence that site-specific screening and the use of sequential extraction procedures are effective methods for evaluating natural attenuation for metals impacted sites.     

Time-dependent observation of individual cellular binding events to field-effect transistors

Electrolyte-gate field-effect transistors (EG-FETs) gained continuously more importance in the field of bioelectronics. The reasons for this are the intrinsic properties of these FETs. Binding of analysts or changes in the electrolyte composition are leading to variations of the drain-source current. Furthermore, due to the signal amplification upon voltage-to-current conversion even small extracellular signals can be detected. Here we report about impedance spectroscopy with an FET array to characterize passive components of a cell attached to the transistor gate. We developed a 16-channel readout system, which provides a simultaneous, lock-in based readout. A test signal of known amplitude and phase was applied via the reference electrode. We monitored the electronic transfer function of the FETs with the attached cell. The resulting frequency spectrum was used to investigate the surface adhesion of individual HEK293 cells. We applied different chemical treatments with either the serinpeptidase trypsin or the ionophor amphotericin B (AmpB). Binding studies can be realized by a time-dependent readout of the lock-in amplifier at a constant frequency. We observed cell detachment upon trypsin activity as well as membrane decomposition induced by AmpB. The results were interpreted in terms of an equivalent electrical circuit model of the complete system. The presented method could in future be applied to monitor more relevant biomedical manipulations of individual cells. Due to the utilization of the silicon technology, our method could be easily up-scaled to many output channels for high throughput pharmacological screening.