Supranucleosomal organization of chromatin

A systematic study of the effect of different ionic conditions on the morphology of the 25–30 nm chromatin fiber from chicken erythrocytes has revealed that, as the ionic strength is increased, knobby fibers with a clear superbead structure are formed in the presence of either Mg⁺⁺ or Na⁺, or both. A further increase in ionic strength results in smooth chromatin fibers due to a tight packing of superbeads. Cross-linking such fibers with formaldehyde and reversal of the ionic conditions, demonstrates the superbead structures underlying the smooth fibers observed at high ionic concentrations. The average size of the superbeads is 34 nm along the length of the fibers, in agreement with the value found in embedded sea cucumber chromatin. A second class of superbeads has an average length of 25 nm and probably corresponds to partially disrupted structures.     

Outer pore topology of the ECaC-TRPV5 channel by cysteine scan mutagenesis

The substituted cysteine accessibility method (SCAM) was used to map the external vestibule and the pore region of the ECaC-TRPV5 calcium-selective channel. Cysteine residues were introduced at 44 positions from the end of S5 (Glu515) to the beginning of S6 (Ala560). Covalent modification by positively charged MTSET applied from the external medium significantly inhibited whole cell currents at 15/44 positions. Strongest inhibition was observed in the S5-linker to pore region (L520C, G521C, and E522C) with either MTSET or MTSES suggesting that these residues were accessible from the external medium. In contrast, the pattern of covalent modification by MTSET for residues between Pro527 and Ile541 was compatible with the presence of a alpha-helix. The absence of modification by the negatively charged MTSES in that region suggests that the pore region has been optimized to favor the entrance of positively charged ions. Cysteine mutants at positions -1, 0, +1, +2 around Asp542 (high Ca2+ affinity site) were non-functional. Whole cell currents of cysteine mutants at +4 and +5 positions were however covalently inhibited by external MTSET and MTSES. Altogether, the pattern of covalent modification by MTS reagents globally supports a KcsA homology-based three-dimensional model whereby the external vestibule in ECaC-TRPV5 encompasses three structural domains consisting of a coiled structure (Glu515 to Tyr526) connected to a small helical segment of 15 amino acids (527PTALFSTFELFLT539) followed by two distinct coiled structures Ile540-Pro544 (selectivity filter) and Ala545-Ile557 before the beginning of S6.     

Estimation of the quantity of bacteria encapsulated in Lentikats Biocatalyst via phospholipid fatty acids content: a preliminary study

The content of phospholipid fatty acids (PLFA) was determined in samples of polyvinyl alcohol lenses (Lentikats Biocatalyst, LB) with encapsulated Paracoccus denitrificans withdrawn during long-term denitrification experiments. The total PLFA content correlated highly with specific denitrification activities of LB as well as biomass estimation based on image analyses of microscopic photos. The results confirmed the applicability of PLFA determination for estimation of the amount of living encapsulated microbial biomass during biotechnological applications.     

Plasmid-encoded lysostaphin endopeptidase gene of Staphylococcus simulans biovar staphylolyticus

A 12.2-kilobase (kb) BclI fragment containing the lysostaphin endopeptidase gene was cloned from Staphylococcus simulans biovar staphylolyticus into Escherichia coli. The gene was expressed in E. coli and the gene product apparently was secreted into the periplasmic space. The gene was localized to a 3.3-kb region of the cloned fragment and this region was shown to contain a staphylococcal promoter for the endopeptidase gene. By hybridization analysis, the endopeptidase gene was shown to reside on the largest of five plasmids in S. simulans biovar staphylolyticus. No additional copies of this gene were detected in the genome.     

Inferring the sources of postglacial range expansion in two large European land snails

Exact locations of glacial refugia are relevant for the study of contemporary biodiversity, not only as places less disturbed during the climatic changes but also as sources of rapid expansion of the biota after the Last Glacial cycle. If continuously inhabited over several of the Quaternary glacial cycles, the refugia are readily identifiable by the accumulated genetic diversity. However, the sources of the Holocene range expansion, particularly important for the emergence of present-day bio- and phylogeographic patterns and for realistic estimation of species’ expansion rates, might have been located at the fringes of the glacial species ranges and lack unique lineages. This problem is pertinent when the variation is explored at slowly evolving genetic markers. We suggest that the location of such source refugia may be approximated by reconstructing the geographic location as a continuous trait evolving along the branches of a phylogenetic tree. We applied this approach, using the BEAST software, on two large southeast European land snail species: Caucasotachea vindobonensis and Helix thessalica. We found evidence for C. vindobonensis refugia in the western Balkans; notable is an apparently old refugium in Bosnia and Herzegovina. The plausible sources of the species’ Holocene range expansion, however, were located around the south-western end of the Carpathians. Although the source areas were likely similar in H. thessalica, some expansion sources suggested by the analyses (e.g., Podolia, Ukraine) appeared implausible and driven by sampling clustered in that area. The applied approach allows for additional exploitation of the mitochondrial data gathered during the past two decades of animal phylogeography studies.     

From Mesolithic hunters to Iron Age herders: a unique record of woodland use from eastern central Europe (Czech Republic)

Vegetation History and Archaeobotany - In a continuous, perfectly stratified sedimentary sequence which was discovered under a large sandstone overhang in northern Bohemia, Czech Republic, we...