Multi-dimensional HPLC/MS of the nucleolar proteome using HPLC-chip/MS

The proteome of the human nucleolus was investigated in a single analysis using off-line strong cation exchange chromatography and microfraction collection combined with HPLC-chip/MS. The analysis was conducted either as a 1-D workflow with HPLC-chip alone or as a 2-D workflow. Two hundred and six unique proteins were identified in the International Protein Index human database corresponding to 2024 unique tryptic peptides identified in the 2-D analysis. In contrast, only 34 proteins and 151 corresponding tryptic peptides were found by applying a 1-D separation strategy. This clearly indicated that the complexity of the samples required the combination of more than one orthogonal separation technique. Stringent database search criteria, including reversal of sequences and therefore better exclusion of false-positive identifications, were applied for reliable protein identification.     

Self-sustained localized structures in a boundary-layer flow

When a boundary layer starts to develop spatially over a flat plate, only disturbances of sufficiently large amplitude survive and trigger turbulence subcritically. Direct numerical simulation of the Blasius boundary-layer flow is carried out to track the dynamics in the region of phase space separating transitional from relaminarizing trajectories. In this intermediate regime, the corresponding disturbance is fully localized and spreads slowly in space. This structure is dominated by a robust pair of low-speed streaks, whose convective instabilities spawn hairpin vortices evolving downstream into transient disturbances. A quasicyclic mechanism for the generation of offspring is unfolded using dynamical rescaling with the local boundary-layer thickness.