Open tubular immobilized metal ion affinity chromatography combined with MALDI MS and MS/MS for identification of protein phosphorylation sites

Protein phosphorylation is one of the most important known posttranslational modifications. Tandem mass spectrometry has become an important tool for mapping out the phosphorylation sites. However, when a peptide generated from the enzymatic or chemical digestion of a phosphoprotein is highly phosphorylated or contains many potential phosphorylation residues, phosphorylation site assignment becomes difficult. Separation and enrichment of phosphopeptides from a digest mixture is desirable and often a critical step for MS/MS-based site determination. In this work, we present a novel open tubular immobilized metal ion affinity chromatography (OT-IMAC) method, which is found to be more effective and reproducible for phosphopeptide enrichment, compared to a commonly used commercial product, Ziptip from Millipore. A strategy based on a combination of OT-IMAC, sequential dual-enzyme digestion, and matrix-assisted laser desorption/ionization (MALDI) quadrupole time-of-flight tandem mass spectrometry for phosphoprotein characterization is presented. It is shown that MALDI MS/MS with collision-induced dissociation can be very effective in generating fragment ion spectra containing rich structural information, which enables the identification of phosphorylation sites even from highly phosphorylated peptides. The applicability of this method for real world applications is demonstrated in the characterization and identification of phosphorylation sites of a Na(+)/H(+) exchanger fusion protein, His182, which was phosphorylated in vitro using the kinase Erk2.     

Passively driven integrated microfluidic system for separation of motile sperm

This paper describes a self-contained integrated microfluidic system that can separate motile sperm from small samples that are difficult to handle using conventional sperm-sorting techniques. The device isolates motile sperm from nonmotile sperm and other cellular debris, based on the ability of motile sperm to cross streamlines in a laminar fluid stream. The device is small, simple, and disposable yet is an integrated system complete with sample inlets, outlets, sorting channel, and a novel passively driven pumping system that provides a steady flow of liquid; it requires no external power source or controls. The device fulfills a need in clinical settings where small amounts of sperm need to be sorted. It also opens the way for convenient bioassays based on sperm motility including at-home motile sperm tests.     

Synergistic science

Should computational materials science be recognized as a field with a role in the community comparable to computational physics or chemistry? With the emergence of multiscale modelling, the answer is a resounding 'yes'.     

Cancellable biometrics and annotations on BioHash

Lately, the once powerful one-factor authentication which is based solely on either password, token or biometric approach, appears to be insufficient in addressing the challenges of identity frauds. For example, the sole biometric approach suffers from the privacy invasion and non-revocable issues. Passwords and tokens are easily forgotten and lost. To address these issues, the notion of cancellable biometrics was introduced to denote biometric templates that can be cancelled and replaced with the inclusion of another independent authentication factor. BioHash is a form of cancellable biometrics which mixes a set of user-specific random vectors with biometric features. In verification setting, BioHash is able to deliver extremely low error rates as compared to the sole biometric approach when a genuine token is used. However, this raises the possibility of two identity theft scenarios: (i) stolen-biometrics, in which an impostor possesses intercepted biometric data of sufficient high quality to be considered genuine and (ii) stolen-token, in which an impostor has access to the genuine token and used by the impostor to claim as the genuine user. We found that the recognition rate for the latter case is poorer. In this paper, the quantised random projection ensemble based on the Johnson–Lindenstrauss Lemma is used to establish the mathematical foundation of BioHash. Based on this model, we elucidate the characteristics of BioHash in pattern recognition as well as security view points and propose new methods to rectify the stolen-token problem.     

Temburong Bridge,Brunei – Design of two cable stayed bridges

The Temburong Bridge Project is a 30 km long dual two‐lane highway crossing over the Brunei Bay. It will connect the relatively isolated Brunei district of Temburong to the other three Brunei districts. The main objective of the project is to stimulate economic growth in the Temburong region by connecting it to the country's airport and ports. The alignment crosses two navigation channels resulting in the need for two cable stayed bridges – the Brunei Channel Bridge (145 m main span) and the Eastern Channel Bridge (260 m main span). The design of these cable stayed bridges is one of the first applications of the Eurocode to a fully concrete cable stayed bridge. Both cable stayed bridges draw on strong Islamic architectural influences from the region to form a tower shape that is unique and instantly recognisable. Temburong Brücke, Brunei – Entwurf von zwei Schrägkabelbrücken Die Temburong Brücke ist eine 30 km lange Verbindung über die Brunei Bucht mit zwei Richtungsfahrbahnen mit je zwei Fahrstreifen. Sie wird den relativ isolierten Distrikt Temburong mit den anderen drei Distrikten Bruneis verbinden. Hauptanliegen des Projekts ist die Stimulierung von ökonomischem Wachstum in der Temburong Region, indem sie an den Flughafen und Seehafen des Landes angebunden wird. Die Trassierung erfordert die Überquerung zweier Schiffahrtsrouten. Dies wird mit der Anordnung zweier Schrägkabelbrücken ermöglicht, der Brunei Channel Brücke (145 m Hauptspannweite) und der Eastern Channel Brücke (260 m Hauptspannweite). Die Bemessung der beiden Brücken stellt eine der ersten Anwendungen des Eurocode auf Schrägkabelbrücken mit Betonüberbau dar. Optisch sind die beiden Brücken geprägt von der islamischen Architektur der Umgebung, die Pylonform ist ein besonderes Erkennungsmerkmal der Brücken.     

Hemi- and Homozygous Loss-of-Function Mutations in DSG2 (Desmoglein-2) Cause Recessive Arrhythmogenic Cardiomyopathy with an Early Onset

About 50% of patients with arrhythmogenic cardiomyopathy (ACM) carry a pathogenic or likely pathogenic mutation in the desmosomal genes. However, there is a significant number of patients without positive familial anamnesis. Therefore, the molecular reasons for ACM in these patients are frequently unknown and a genetic contribution might be underestimated. Here, we used a next-generation sequencing (NGS) approach and in addition single nucleotide polymor-phism (SNP) arrays for the genetic analysis of two independent index patients without familial medical history. Of note, this genetic strategy revealed a homozygous splice site mutation (DSG2–c.378+1G>T) in the first patient and a nonsense mutation (DSG2–p.L772X) in combination with a large deletion in DSG2 in the second one. In conclusion, a recessive inheritance pattern is likely for both cases, which might contribute to the hidden medical history in both families. This is the first report about these novel loss-of-function mutations in DSG2 that have not been previously identi-fied. Therefore, we suggest performing deep genetic analyses using NGS in combination with SNP arrays also for ACM index patients without obvious familial medical history. In the future, this finding might has relevance for the genetic counseling of similar cases.