Two episodes of vancomycin-resistant Enterococcus faecium outbreaks caused by two genetically different clones in a newborn intensive care unit.

In 2001 two outbreak episodes (January-March and June-July) caused by vancomycin-resistant E. faecium (VRE) of the VanA-type were observed at a neonatal intensive care unit (NICU) of a university hospital in south-west Germany. To identify the initial source and the route of transmission environmental samples were examined as well as stool samples from patients and the staff. VRE was not found in environmental samples. However, stool samples from 24 hospitalised children tested positive and bacterial clonality was assessed by Sma1-based macro restriction analysis. Furthermore, esp gene and vancomycin resistance gene carriage were examined as well as bacteriocin production. PCR analysis showed that all 24 isolates carried vanA gene cluster, encoding resistance to vancomycin and teicoplanin. However, five of the vanA-positive isolates were resistant to vancomycin but not to teicoplanin. Only these five isolates produced bacteriocin, but in none of the isolates esp gene was detected. PFGE revealed that both outbreaks were caused by two different clones. The patient initiating the first episode, was identified whereas the origin of the second episode remained unknown. From one of the 40 staff stool samples VRE was isolated. This strain was related to the clone of the summer outbreak. In conclusion there were two independent episodes of self limiting VRE outbreaks and transmission on the ward is highly probable.

DAP-like kinase, a member of the death-associated protein kinase family, associates with centrosomes, centromers, and the contractile ring during mitosis.

DAP-like kinase (Dlk) is a nuclear serine/threonine-specific kinase which has been implicated in apoptosis. However, induction of apoptosis by Dlk requires its relocation to the cytoplasm, particularly association with the actin cytoskeleton, which is achieved through interaction with pro-apoptotic protein Par-4. On the other hand, nuclear Dlk does not induce apoptosis and has rather been implicated in transcription. To further explore the biological functions of Dlk, we established a cell clone of MCF-7 cells stably expressing a GFP-Dlk fusion protein at low level. Ectopic expression of GFP-Dlk did not affect the growth properties of the cells. During interphase, GFP-Dlk showed a diffuse nuclear distribution with punctate staining in a subpopulation of cells. During mitosis, however, Dlk was associated with centrosomes, centromeres, and the contractile ring, but not with the mitotic spindle. Association with centrosomes, as confirmed by colocalization with gamma-tubulin and pericentrin persisted throughout mitosis but was also seen in interphase cells. Interestingly, GFP-Dlk and gamma-tubulin could be co-immunoprecipitated indicating that they are present in the same protein complex. Association of Dlk with centromeres, as verified by confocal fluorescence microscopy with centromere-specific antibodies was more restricted and discernable from prophase to early anaphase. Centromere association of Dlk coincides with H3 phosphorylation at Thr11 that is specifically phosphorylated by Dlk in vitro (U. Preuss, G. Landsberg, K. H. Scheidtmann, Nucleic Acids Res. 31, 878-885, 2003). During cytokinesis, Dlk was enriched in the contractile acto-myosin ring and colocalized with Ser19-phosphorylated myosin light chain, which is an in vitro substrate of Dlk. Strikingly, a C-terminal truncation mutant of Dlk generated multi-nucleated cells. Together, these data suggest that Dlk participates in regulation and, perhaps, coordination of mitotis and cytokinesis.