The taxonomic status of two actinomycetes isolated from the wall of a hypogean Roman catacomb was established based on a polyphasic investigation. The organisms were found to have chemical and morphological markers typical of members of the genus Amycolatopsis. They also shared a range of chemical, molecular and phenotypic markers which served to separate them from representatives of recognized Amycolatopsis species. The new isolates formed a branch in the Amycolatopsis 16S rRNA gene sequence tree with Amycolatopsis minnesotensis NRRL B-24435 T , but this association was not supported by a particularly high bootstrap value or by the product of the maximum-parsimony tree-making algorithm. The organisms were distinguished readily from closely related Amycolatopsis species based on a combination of phenotypic properties and from all Amycolatopsis strains by their characteristic menaquinone profiles, in which tetra-hydrogenated menaquinones with 11 isoprene units predominated. The combined genotypic and phenotypic data indicate that the isolates merit recognition as representing a novel species of the genus Amycolatopsis. The name proposed for this novel species is Amycolatopsis nigrescens sp. nov., with type strain CSC17Ta-90 T (=HKI 0330 T =DSM 44992 T =NRRL B-24473 T ).The genus Amycolatopsis Lechevalier et al. 1986 is a member of the family Pseudonocardiaceae Embley et al. 1988 (as circumscribed by Warwick et al., 1994). Members of this family are characterized by the presence of meso-diaminopimelic acid, arabinose and galactose in whole-cell hydrolysates (wall chemotype IV sensu Lechevalier & Lechevalier, 1970), fatty acids that mainly consist of iso-and anteisobranched components and N-acetylated muramic acid and by the lack of mycolic acids (Takahashi, 2001). Amycolatopsis strains can be separated from members of the other genera classified within the family Pseudonocardiaceae by using a range of chemotaxonomic and morphological markers (Kim & Goodfellow, 1999) and by genus-specific oligonucleotide primers based on 16S rRNA gene sequences (Tan et al., 2006).Members of the genus Amycolatopsis are Gram-positive, non-acid-fast, non-motile actinomycetes that form branched vegetative hyphae that undergo fragmentation into rod-like and squarish elements. If present, aerial hyphae may be sterile or differentiate into squarish to ovoid sporelike structures (Chun et al., 1999). The genus encompasses alkaliphilic, mesophilic, thermophilic and pathogenic species, which can be distinguished from one another by using a range of phenotypic properties (SaintpierreBonaccio et al., 2005;Lee, 2006;Lee et al., 2006). At the time of writing, the genus Amycolatopsis comprised 32 recognized species and subspecies, although there is compelling evidence that the genus is grossly underspeciated and that Amycolatopsis fastidiosa should be classified within a different taxon (Tan et al., 2006). The present polyphasic study was designed to determine the taxonomic position of two Amycolatopsis-like strains that had been isolated from a h...

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Coxsackievirus B3 (CVB3) is a common factor in human myocarditis. The interplay between host factors and virus components is crucial for the fate of the infected cells. Despite that, host protein responses, which characterize CVB3-induced diseases, have not yet been determined in detail. To investigate the nature of modified protein patterns in infected human cells compared with uninfected cells, two-dimensional gel electrophoresis in combination with matrix-assisted laser desorption/ionization-mass spectrometry were used. The regulated proteins, e.g. nucleophosmin (nucleolar protein B23), lamin, the RNA-binding protein UNR and the p38 mitogen-activated protein kinase, were sorted according to their functional groups and interpreted in the context of the myocarditis process.Coxsackievirus B3 (CVB3), a member of the family Picornaviridae, is an important human pathogen that has been associated with serious diseases. Clinically, coxsackievirus infections are known to be associated with different forms of subacute, acute and chronic myocarditis as well as pancreatitis (Baboonian et al., 1997;Reyes & Lerner, 1985;Woodruff, 1980). CVB3 may cause cardiac arrhythmias and acute heart failure; chronic forms of this disease may occur, leading to dilated cardiomyopathy (DCM) (Frisk et al., 1984;Kandolf, 1998;Kandolf & Hofschneider, 1989). DCM results in a progressive heart insufficiency and a significantly increased death rate (Gillum, 1986;Sugrue et al., 1992). The pathogenesis of coxsackievirus infection has been studied extensively in different murine models, demonstrating that the outcome of this viral infection is determined by complex interactions among several variables of the virus and the host (Chow et al., 1991;Huber, 1997). In addition, the mechanisms for how CVB3 causes myocarditis are not very well characterized (Bowles & Towbin, 1998).The host elements responsible for the changes observed during the course of CVB3-mediated myocarditis have not yet been determined intensively. It is well documented that the picornavirus proteases 2A and 3C cleave cellular proteins, e.g. the translation initiation factor eIF4G (Haghighat et al., 1996) or the poly(A)-binding protein (Kerekatte et al., 1999), resulting in a host translation shut-off. However, not much is known about cellular proteins that are modified during the infection process and which are necessary furthermore for the virus to replicate. Previously, we demonstrated a direct interaction between the CVB3 capsid protein VP2 and the human protein Siva, overexpressed upon a CVB3 infection (Henke et al., 2000(Henke et al., , 2001, which is involved in the CD27/CD70-transduced apoptotic pathway (Prasad et al., 1997). To expand rapidly the portrait of host gene expression involved in the pathogenesis of viral myocarditis and particularly to examine the expression of proteins, we used a proteome-wide approach. Proteins of infected and non-infected HeLa cells as well as HepG2 cells were separated on two-dimensional (2D) gels and spots were analysed by peptide mass fi...

The identification of different Kitasatospora strains has been shown with a DNA-chip based on an electrical readout scheme. The 16S-23S rDNA internal transcribed spacer region of these Actinomycetes was used for identification. Two different capture probes per strain were immobilized on the chip. The capture probes were spotted on a DNA-chip with electrode structures for an electrical DNA detection. A biotinylated PCR product of the 16S-23S rDNA region was incubated on the chips and bound to its complementary capture sequences. Followed by a gold nanoparticle or enzyme labeling and a deposition of silver, the binding of the PCR product was detected by an increase of the measured conductivity on the chip. To show the applicability of this detection system, four strains of Kitasatospora were chosen for an identification using the DNA-chip with electrical detection. Each strain was clearly identified using the system. Concentrations of the polymerase chain reaction (PCR) products within the range of 1 ng/ml to 1 mug/ml were detected and identified. These tests are the first application of this novel electrical detection scheme for the identification and classification of microorganisms. The presented results show that the DNA-chip with electrical detection can be used for a robust and cost-efficient DNA analysis.

Three actinomycetes (strains HKI 0478 T , HKI 0479 and HKI 0480) isolated from the surfaces of rocks in the Feengrotten medieval alum slate mine (Thuringia, Germany) were examined in a polyphasic taxonomic study. The following morphological and chemotaxonomic features supported their classification as members of the genus Kribbella: the presence of LLdiaminopimelic acid in the cell-wall peptidoglycan; glucose together with minor amounts of mannose and ribose as the whole-cell sugars; polar lipids comprising phosphatidylcholine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and unknown phospho-and glycolipids; fatty acid profiles characterized by the predominance of anteiso-C 15 : 0 , iso-C 16 : 0 and C 16 : 0 9-methyl; and the presence of MK-9(H 4 ) as the main menaquinone. The isolates had almost identical 16S rRNA gene sequences (99.9-100 %) and were most closely related to the type strains of Kribbella jejuensis (98.9 % sequence similarity), Kribbella swartbergensis and Kribbella solani (both 98.8 %). A wide range of genotypic and phenotypic markers as well as the low levels of DNA-DNA relatedness between strain HKI 0478 T and the type strains of K. jejuensis (41.3 %), K. swartbergensis (18.6 %) and K. solani (14.2 %) distinguished the novel strains from their closest phylogenetic neighbours. On the basis of these results, strain HKI 0478 T represents a novel member of the genus Kribbella, for which the name Kribbella aluminosa sp. nov. is proposed. The type strain is HKI 0478 T (5DSM 18824 T 5JCM 14599 T ). Park et al. 1999 emend. Sohn et al. 2003 is a member of the family Nocardioidaceae, which was proposed by Nesterenko et al. (1985Nesterenko et al. ( , 1990 and the description of which was emended by Rainey et al. (in Stackebrandt et al., 1997). The genus Kribbella is represented by Gram-positive or Gram-variable, nonmotile actinomycetes that form an extensively branched vegetative mycelium and aerial hyphae that fragment into short to elongated rod-like or coccoid elements. Strains of the 11 Kribbella species with validly published names share the following chemotaxonomic characteristics (Park et al., 1999;Sohn et al., 2003): the presence of LL-diaminopimelic acid in the peptidoglycan (wall chemotype I sensu Lechevalier & Lechevalier, 1970), fatty acids that consist mainly of anteiso-and iso-branched components, MK-9(H 4 ) as the main menaquinone and polar lipids comprising phosphatidylcholine, diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol (phospholipid pattern III sensu Lechevalier et al., 1977). A combination of these chemotaxonomic markers serves to separate the strains of the genus Kribbella from members of the other genera within the family Nocardioidaceae. The genus KribbellaThree filamentously growing actinomycetes, strains HKI 0478 T , HKI 0479 and HKI 0480, were isolated from acidic and heavy-metal-containing rock surfaces in a small mining area behind the 'Märchendom', the third level of the Feengrotten medieval alum slate mine (Saalfeld, Thuringia,...