High lateral resolution vs molecular preservation in near-IR fs-laser desorption postionization mass spectrometry.

Ultrashort pulse length lasers operating in the near-infrared region show promise for submicrometer lateral resolution by laser desorption-based mass spectrometry (MS) imaging. However, these experiments must balance lateral resolution and molecular fragmentation since abundant atomic ions are observed at the high laser irradiances that can be generated by tightly focused ultrashort pulse laser beams. It is shown here that combining ultrashort pulse laser desorption with laser postionization (fs-LDPI) allows for a considerable increase of molecular ion signal while operating with lower laser irradiances, yielding the added benefit of reduced molecular fragmentation. This Letter presents several experimental results in support of the fs-LDPI approach for MS imaging. First, the lateral resolution for MS imaging of molecular species desorbed by ∼75 fs, 800 nm laser pulses was determined to be <2 µm for a simulated organic electronic device under vacuum. Next, the dependence of precursor ion survival on both desorption laser fluence and delay between desorption and photoionization laser pulses was observed for a small molecule desorbed from an organic multilayer that was originally devised as a model of a bacterial biofilm. When considered in light of recent results in the literature (Milasinovic et al. J. Phys. Chem. C 2014, DOI: 10.1021/jp504062u), these experiments demonstrate the potential for submicrometer spatial resolution MS imaging by fs-LDPI.

Multiple elements regulate nuclear/cytoplasmic shuttling of FOXO1: characterization of phosphorylation- and 14-3-3-dependent and -independent mechanisms.

FOXO1, a Forkhead transcription factor, is an important target of insulin and growth factor action. Phosphorylation of Thr-24, Ser-256 and Ser-319 promotes nuclear exclusion of FOXO1, yet the mechanisms regulating nuclear/cytoplasmic shuttling of FOXO1 are poorly understood. Previous studies have identified an NLS (nuclear localization signal) in the C-terminal basic region of the DBD (DNA-binding domain), and a leucine-rich, leptomycin-B sensitive NES (nuclear export signal) located further downstream. Here, we find that other elements in the DBD also contribute to nuclear localization, and that multiple mechanisms contribute to nuclear exclusion of FOXO1. Phosphorylation of Ser-319 and a cluster of nearby residues (Ser-322, Ser-325 and Ser-329) functions co-operatively with the nearby NES to promote nuclear exclusion. The N-terminal region of FOXO1 (amino acids 1-149) also is sufficient to promote nuclear exclusion, and does so through multiple mechanisms. Amino acids 1-50 are sufficient to promote nuclear exclusion of green fluorescent protein fusion proteins, and the phosphorylation of Thr-24 is required for this effect. A leucine-rich, leptomycin B-sensitive export signal is also present nearby. Phosphorylated FOXO1 binds 14-3-3 proteins, and co-precipitation studies with tagged proteins indicate that 14-3-3 binding involves co-operative interactions with both Thr-24 and Ser-256. Ser-256 is located in the C-terminal region of the DBD, where 14-3-3 proteins may interfere both with DNA-binding and with nuclear-localization functions. Together, these studies demonstrate that multiple elements contribute to nuclear/cytoplasmic shuttling of FOXO1, and that phosphorylation and 14-3-3 binding regulate the cellular distribution and function of FOXO1 through multiple mechanisms. The presence of these redundant mechanisms supports the concept that the regulation of FOXO1 function plays a critical role in insulin and growth factor action.

Cluster beam deposition of Cu(2-X)S nanoparticles into organic thin films.

Bulk-heterojunction films composed of semiconductor nanoparticles blended with organic oligomers are of interest for photovoltaic and other applications. Cu2-XS nanoparticles were cluster beam deposited into thermally evaporated pentacene or quaterthiophene to create bulk-heterojunction thin films. The nanoparticle stoichiometry, morphology, and chemistry within these all-gas phase deposited films were characterized by X-ray photoelectron spectroscopy (XPS) and electron microscopy. Cu2-XS nanoparticles were (at most) only slightly copper-deficient with respect to Cu2S; ∼2.5 nm diameter, unoxidized Cu2-XS nanoparticles formed in both pentacene and quaterthiophene, as the matrix was not observed to impact the nanoparticle morphology or chemical structure. Cluster beam deposition allowed direct control of the nanoparticle stoichiometry and nanoparticle:organic ratio. Chemical states or Wagner plots were combined with other XPS data analysis strategies to determine the metal oxidation state, indicating that Cu(I) was predominant over Cu(II) in the Cu2-XS nanoparticles.