Accelerated Barocycler Lysis and Extraction Sample Preparation for Clinical Proteomics by Mass Spectrometry.

We have developed a streamlined proteomic sample preparation protocol termed Accelerated Barocycler Lysis and Extraction (ABLE) that substantially reduces the time and cost of tissue sample processing. ABLE is based on pressure cycling technology (PCT) for rapid tissue solubilization and reliable, controlled proteolytic digestion. Here, a previously reported PCT based protocol was optimized using 1-4 mg biopsy punches from rat kidney. The tissue denaturant urea was substituted with a combination of sodium deoxycholate (SDC) and N-propanol. ABLE produced comparable numbers of protein identifications in half the sample preparation time, being ready for MS injection in 3 h compared with 6 h for the conventional urea based method. To validate ABLE, it was applied to a diverse range of rat tissues (kidney, lung, muscle, brain, testis), human HEK 293 cell lines, and human ovarian cancer samples, followed by SWATH-mass spectrometry (SWATH-MS). There were similar numbers of quantified proteins between ABLE-SWATH and the conventional method, with greater than 70% overlap for all sample types, except muscle (58%). The ABLE protocol offers a standardized, high-throughput, efficient, and reproducible proteomic preparation method that when coupled with SWATH-MS has the potential to accelerate proteomics analysis to achieve a clinically relevant turn-around time.

Protease sensing using nontoxic silicon quantum dots.

Herein is presented a proof-of-concept study of protease sensing that combines nontoxic silicon quantum dots (SiQDs) with Förster resonance energy transfer (FRET). The SiQDs serve as the donor and an organic dye as the acceptor. The dye is covalently attached to the SiQDs using a peptide linker. Enzymatic cleavage of the peptide leads to changes in FRET efficiency. The combination of interfacial design and optical imaging presented in this work opens opportunities for use of nontoxic SiQDs relevant to intracellular sensing and imaging.

Controlling self-assembly of diphenylalanine peptides at high pH using heterocyclic capping groups.

Using small angle neutron scattering (SANS), it is shown that the existence of pre-assembled structures at high pH for a capped diphenylalanine hydrogel is controlled by the selection of N-terminal heterocyclic capping group, namely indole or carbazole. At high pH, changing from a somewhat hydrophilic indole capping group to a more hydrophobic carbazole capping group results in a shift from a high proportion of monomers to self-assembled fibers or wormlike micelles. The presence of these different self-assembled structures at high pH is confirmed through NMR and circular dichroism spectroscopy, scanning probe microscopy and cryogenic transmission electron microscopy.

A Surgical Business Composite Score for Army Medicine.

Measuring surgical business performance for Army military treatment facilities is currently done through 6 business metrics developed by the Army Medical Command (MEDCOM) Surgical Services Service Line (3SL). Development of a composite score for business performance has the potential to simplify and synthesize measurement, improving focus for strategic goal setting and implementation. However, several considerations, ranging from data availability to submetric selection, must be addressed to ensure the score is accurate and representative. This article presents the methodology used in the composite score's creation and presents a metric based on return on investment and a measure of cases recaptured from private networks.

Biocompatible small peptide super-hydrogelators bearing carbazole functionalities.

For the first time we have introduced carbazole capping groups onto two short peptide sequences, namely a diphenylalanine dipeptide and a glycine-diphenylalanine tripeptide, giving compounds 1 and 2, respectively. Both molecules form hydrogels at low concentrations, as low as 0.03% (w/v) for 1- well within the range of supergelators. Both gelators are composed of small ca. 2 nm thick molecular fibres, as elucidated by AFM and are non-cytotoxic towards HeLa cells at concentrations which are at or above their minimum gelation concentration.

Exceptionally strong hydrogels through self-assembly of an indole-capped dipeptide.

The synthesis of a new hydrogelator with an indole capping group, 1, is reported. 1 forms exceptionally strong hydrogels in a variety of environments, with values for the storage modulus G' amongst the highest reported for supramolecular hydrogels. These gels exhibit strong bundling characteristics, which gives the high values for G' observed. Cell viability studies show that at low concentrations, 1 is biocompatible, however upon self-assembly at higher concentrations, cytotoxic effects are observed.