Orthogonal Translation Meets Electron Transfer: In Vivo Labeling of Cytochrome c for Probing Local Electric Fields

Cytochrome c (cyt c), a redox protein involved in diverse fundamental biological processes, is among the most traditional model proteins for analyzing biological electron transfer and protein dynamics both in solution and at membranes. Studying the role of electric fields in energy transduction mediated by cyt c relies upon appropriate reporter groups. Up to now these had to be introduced into cyt c by in vitro chemical modification. Here, we have overcome this restriction by incorporating the noncanonical amino acid p‐cyanophenylalanine (pCNF) into cyt c in vivo. UV and CD spectroscopy indicate preservation of the overall protein fold, stability, and heme coordination, whereas a small shift of the redox potential was observed by cyclic voltammetry. The C≡N stretching mode of the incorporated pCNF detected in the IR spectra reveals a surprising difference, which is related to the oxidation state of the heme iron, thus indicating high sensitivity to changes in the electrostatics of cyt c.     

Structural invariance and age-related performance differences in face cognition.

Perceiving and memorizing faces swiftly and correctly are important social competencies. The organization of these interpersonal abilities and how they change across the life span are still poorly understood. We investigated changes in the mean and covariance structure of face cognition abilities across the adult life span. A sample of 448 participants, with age ranging from 18 to 88 years, completed a battery of 15 face cognition tasks. After establishing a measurement model of face cognition that distinguishes between face perception, face memory, and the speed of face cognition, we used multiple group models and age-weighted measurement models to explore age-related changes. The modeling showed that the loadings and intercepts of all measures are age invariant. The factor means showed substantial decrements with increasing age. Age-related decrements in performance were strongest for the speed of face cognition but were also salient for face perception and face memory. The onset of age decrements is apparent in the 60s for face perception, in the late 40s for face memory, and in the early 30s for speed of face cognition. Implications of these findings on a theoretical and methodological level are discussed, and potential consequences for applied settings are considered. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

MBE P-on-n Hg1−xCdxTe heterostructure detectors on silicon substrates

The capability of growing state-of-the-art middle wavelength infrared (MWIR)-HgCdTe layers by molecular beam epitaxy (MBE) on large area silicon substrates has been demonstrated. We have obtained excellent compositional uniformity with standard deviation of 0.001 with mean composition of 0.321 across 1.5″ radii. R₀A as high as 5 × 10⁷ ω-cm² with a mean value of 7 × 10⁶ Θ-cm² was measured for cut-off wavelength of 4.8 μm at 77K. Devices exhibit diffusion limited performance for temperatures above 95K. Quantum efficiencies up to 63% were observed (with no anti-reflection coating) for cut-off wavelength (4.8–5.4) μm @ 77K. Excellent performance of the fabricated photodiodes on MBE HgCdTe/CdTe/Si reflects on the overall quality of the grown material in the MWIR region.     

Nanobodies and Nanocrystals: Highly Sensitive Quantum Dot‐Based Homogeneous FRET Immunoassay for Serum‐Based EGFR Detection

Semiconductor quantum dot nanocrystals (QDs) for optical biosensing applications often contain thick polyethylene glycol (PEG)‐based coatings in order to retain the advantageous QD properties in biological media such as blood, serum or plasma. On the other hand, the application of QDs in Förster resonance energy transfer (FRET) immunoassays, one of the most sensitive and most common fluorescence‐based techniques for non‐competitive homogeneous biomarker diagnostics, is limited by such thick coatings due to the increased donor‐acceptor distance. In particular, the combination with large IgG antibodies usually leads to distances well beyond the common FRET range of approximately 1 to 10 nm. Herein, time‐gated detection of Tb‐to‐QD FRET for background suppression and an increased FRET range is combined with single domain antibodies (or nanobodies) for a reduced distance in order to realize highly sensitive QD‐based FRET immunoassays. The “(nano)²” immunoassay (combination of nanocrystals and nanobodies) is performed on a commercial clinical fluorescence plate reader and provides sub‐nanomolar (few ng/mL) detection limits of soluble epidermal growth factor receptor (EGFR) in 50 μL buffer or serum samples. Apart from the first demonstration of using nanobodies for FRET‐based immunoassays, the extremely low and clinically relevant detection limits of EGFR demonstrate the direct applicability of the (nano)^2‐ assay to fast and sensitive biomarker detection in clinical diagnostics.     

Construction of a Fuzzy and Boolean Logic Gates Based on DNA

Logic gates are devices that can perform logical operations by transforming a set of inputs into a predictable single detectable output. The hybridization properties, structure, and function of nucleic acids can be used to make DNA‐based logic gates. These devices are important modules in molecular computing and biosensing. The ideal logic gate system should provide a wide selection of logical operations, and be integrable in multiple copies into more complex structures. Here we show the successful construction of a small DNA‐based logic gate complex that produces fluorescent outputs corresponding to the operation of the six Boolean logic gates AND, NAND, OR, NOR, XOR, and XNOR. The logic gate complex is shown to work also when implemented in a three‐dimensional DNA origami box structure, where it controlled the position of the lid in a closed or open position. Implementation of multiple microRNA sensitive DNA locks on one DNA origami box structure enabled fuzzy logical operation that allows biosensing of complex molecular signals. Integrating logic gates with DNA origami systems opens a vast avenue to applications in the fields of nanomedicine for diagnostics and therapeutics.     

Layered Perovskite Nanofibers via Electrospinning for Overall Water Splitting

The (111)‐layered perovskite materials Ba₅Ta₄O₁₅, Ba₅Ta₂Nb₂O₁₅ and Ba₅Nb₄O₁₅ are prepared with nanofiber morphology via electrospinning for the first time. The nanofibers are built up from small single crystals, with up to several micrometers length even after calcination. The formation mechanism is investigated in detail, revealing an intermediate formation of amorphous barium carbonate strengthening the nanofiber morphology for high temperature treatment. All nanofiber compounds are able to generate hydrogen without any co‐catalyst in photocatalytic reformation of methanol. After photodeposition of Rh‐Cr₂O₃ co‐catalysts, the nanofibers show better activity in overall water splitting compared to sol–gel‐derived powders.     

Characterization of the major bovine brain Go alpha isoforms. Mapping the structural differences between the alpha subunit isoforms identifies a variable region of the protein involved in receptor interactions

Go is the major G protein in bovine brain, with at least three isoforms, GoA, GoB, and GoC. Whereas alphaoA and alphaoB arise from a single Goalpha gene as alternatively spliced mRNAs, alphaoA and alphaoC are thought to differ by covalent modification. To test the hypothesis that alphaoA and alphaoC have different N-terminal lipid modifications, proteolytic fragments of alphao isoforms were immunoprecipitated with an N terminus-specific antibody and analyzed by matrix-assisted laser desorption ionization mass spectrometry. The major masses observed in immunoprecipitates were the same for all three alphao isoforms and corresponded to the predicted mass of a myristoylated N-terminal fragment. Structural differences between alphaoA and alphaoC were also compared before and after limited tryptic proteolysis using SDS-polyacrylamide gel electrophoresis containing 6 M urea. Based upon the alphao subunit fragments produced under activating and nonactivating conditions, differences between alphaoA and alphaoC were localized to a C-terminal fragment of the protein. This region, involved in receptor and effector interactions, implies divergent signaling roles for these two alphao proteins. Finally, the structural difference between alphaoA and alphaoC is associated with a difference of at most 2 daltons based upon measurements by electrospay ionization mass spectrometry.     

Autosomal dominant medullary cystic kidney disease: evidence of gene locus heterogeneity

Autosomal dominant medullary cystic kidney disease (ADMCKD; synonym: medullary cystic disease, MCD) is an autosomal dominant kidney disorder, sharing morphological and clinical features with recessive juvenile nephronophthisis (NPH), such as reduced urinary concentration ability and multiple renal cysts at the corticomedullary junction. While in NPH end-stage renal disease (ESRD) occurs in adolescence, ADMCKD leads to ESRD in adulthood. Recently a gene locus for ADMCKD has been localized to chromosome 1q21 in two large Cypriot families. This prompted us to examine linkage in three ADMCKD-families, using the same set of polymorphic microsatellite markers spanning the critical region on chromosome 1q21. Haplotype analysis revealed that none of the three families showed linkage to this locus, thus demonstrating evidence for genetic locus heterogeneity. Additional linkage analysis studies need to be performed in order to identify further gene loci cosegregating with this autosomal dominant kidney disorder.