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Marine mammals are an important part of ocean ecosystems, of which, whales play a vital role in the marine food chain. In this study, the mucosa and contents from different intestinal tract segments (ITSs) of a stranded dwarf minke whale (Balaenoptera acutorostrata) were analyzed. The gut microbiota were sequenced using high‐throughput sequencing technology, based on a 16S rRNA approach. The microbial composition of the intestinal mucosa and its contents were similar in every single ITS. Large intestine microbiota richness and diversity were significantly higher when compared to the duodenum and jejunum. The dominant bacteria in the gut were Firmicutes and Actinobacteria; the former was enriched in the large intestine, whereas the latter was more abundant in the duodenum and jejunum. Our findings provide novel insights for microbiota in B. acutorostrata., This study has characterized the microbiota of five sampling sources (duodenum, jejunum, cecum, colon, and rectum) in a fresh stranded Balaenoptera acutorostrata carcass. Significant differences were observed in gut microbiota composition across different intestinal tract segments; the microbiomes of posterior compartments were more diverse than the foregut. The duodenum and jejunum were enriched for Erysipelotrichales, Selenomonadales, and Coriobacteriales, whereas Clostridiales were more abundant in the large intestine.

Rationale Bottom-up tandem mass spectrometry (MS/MS) is regularly used in proteomics to identify proteins from a sequence database. De novo sequencing is also available for sequencing peptides with relatively short sequence lengths. We recently showed that paired Lys-C and Lys-N proteases produce peptides of identical mass and similar retention time, but different tandem mass spectra. Such parallel experiments provide complementary information, and allow for up to 100% MS/MS sequence coverage. Methods Here, we report digestion by paired Lys-C and Lys-N proteases of a seven-protein mixture: human hemoglobin alpha, bovine carbonic anhydrase 2, horse skeletal muscle myoglobin, hen egg white lysozyme, bovine pancreatic ribonuclease, bovine rhodanese, and bovine serum albumin, followed by reversed-phase nanoflow liquid chromatography, collision-induced dissociation, and 14.5 T Fourier transform ion cyclotron resonance mass spectrometry. Results Matched pairs of product peptide ions of equal precursor mass and similar retention times from each digestion are compared, leveraging single-residue transposed information with independent interferences to confidently identify fragment ion types, residues, and peptides. Selected pairs of product ion mass spectra for de novo sequenced protein segments from each member of the mixture are presented. Conclusions Pairs of the transposed product ions as well as complementary information from the parallel experiments allow for both high MS/MS coverage for long peptide sequences and high confidence in the amino acid identification. Moreover, the parallel experiments in the de novo sequencing reduce false-positive matches of product ions from the single-residue transposed peptides from the same segment, and thereby further improve the confidence in protein identification. Copyright © 2016 John Wiley & Sons, Ltd.

Chromium nitride (CrN) coatings with different steps of intermediate argon plasma treatments were deposited with primary (200) orientation by multi-arc ion plating technique. By virtue of scanning electron microscopy, X-ray diffraction and high-resolution transmission electron microscopy, the influence of intermediate argon plasma treatments on the coating microstructures, mechanical properties and corrosion properties as well as tribological behaviors in artificial seawater solutions were systematically investigated. It was assumed that the mechanical properties, adhesion strength, corrosion and tribological performances of coatings depended on argon plasma treatment steps. High-performance coatings could be obtained by proper plasma treatment steps. The superior anti-corrosion ability of coating with appropriate treatment steps may be ascribed to the increased charge transfer resistance due to alternative interface and CrN layer and the compact microstructure. On the other hand, the excellent tribological performances in seawater conditions may be attributed to the enhanced mechanical properties. Otherwise, further increase in treatment steps was assumed to distinctly increase defects and deteriorate the coating integrity thus weakening coating properties and behaviors. Copyright © 2016 John Wiley & Sons, Ltd.

The potential epitope of a recombinant food allergen protein, cashew Ana o 1, reactive to monoclonal antibody, mAb 2G4, has been mapped by solution-phase amide backbone H/D exchange (HDX) monitored by Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Purified mAb 2G4 was incubated with recombinant Ana o 1 (rAna o 1) to form antigen:monoclonal antibody (Ag:mAb) complexes. Complexed and uncomplexed (free) rAna o 1 were then subjected to HDX-MS analysis. Five regions protected from H/D exchange upon mAb binding are identified as potential conformational epitope-contributing segments.

RATIONALE Paired Lys-N and Lys-C proteases produce peptides of identical mass and similar retention time, but different tandem mass spectra. Data from these parallel experiments provide constraints that are applied before data analysis. With this approach, we can find matched spectra before analysis, distinguish ion type, and determine residue level confidence. METHODS Aliquots are digested separately by Lys-N and Lys-C peptidases, and analyzed by reversed-phase nano-flow liquid chromatography, collision-induced dissociation, and 14.5 T Fourier transform ion cyclotron resonance mass spectrometry. Matched pairs of fragmentation spectra with equal precursor mass and similar retention times from each digestion are compared, leveraging single-residue transposed information with independent interferences to confidently identify fragment ion type, residues, and peptides. The paired spectra are solved together as a single de novo sequencing problem. RESULTS Two pairs of spectra of a de novo sequenced 18-mer are presented. In one example, the 18-mer has coverage of all residues except the N- and C- terminal lysines and their adjacent residues. The confidence level is high due to six pairs of transposed ions. In the other example, the coverage is incomplete. Nonetheless, nine pairs of transposed ions facilitate identification of two trimer sequence tags with high confidence, one with medium confidence, and additional sequence information with residue-by-residue confidence, thus demonstrating the value of residue-by-residue confidence. CONCLUSIONS Sequence identity and variability, such as post-translational modifications (PTMs), are essential to understanding biological function and disease. The present method facilitates discovery of new peptides with multiple levels of confidence, promises potential characterization of PTMs, and validates peptides from databases. Independent validation may be of interest for a number of applications. Copyright © 2015 John Wiley & Sons, Ltd.

The regeneration potential of mesenchymal stem cells (MSCs) diminishes with advanced age and this diminished potential is associated with changes in cellular functions. This study compared MSCs isolated from the bone marrow of rhesus monkeys (rBMSCs) in three age groups: young ( 12 years). The effects of aging on stem cell properties and indicators of stem cell fitness such as proliferation, differentiation, circadian rhythms, stress response proteins, miRNA expression, and global histone modifications in rBMSCs were analyzed. rBMSCs demonstrated decreased capacities for proliferation and differentiation as a function of age. The production of heat shock protein 70 (HSP70) and heat shock factor 1 (HSF1) were also reduced with increasing age. The level of a core circadian protein, Rev-erb α, was significantly increased in rBMSCs from old animals. Furthermore, analysis of miRNA expression profiles revealed an up-regulation of mir-766 and mir-558 and a down-regulation of mir-let-7f, mir-125b, mir-222, mir-199-3p, mir-23a, and mir-221 in old rBMSCs compare to young rBMSCs. However, there were no significant age-related changes in the global histone modification profiles of the four histone core proteins: H2A, H2B, H3, and H4 on rBMSCs. These changes represent novel insights into the aging process and could have implications regarding the potential for autologous stem cells therapy in older patients.