Synthesis and monolayer characterization of phosphorylated amino acid analogs

The synthesis of a series of thiols containing phosphorylated and non-phosphorylated serine, threonine, and tyrosine amino acid residues is described. The synthesized molecules, based on 3-mercaptopropionic acid, were assembled onto gold and subsequently characterized using infrared reflection-absorption spectroscopy, ellipsometry, X-ray photoelectron spectroscopy, and contact angle goniometry. The ellipsometric analysis indicates that they form densely packed and well-oriented monolayers on gold, with thicknesses that are in good agreement with estimated values from space-filling models. The bulky and space-demanding phosphorylated threonine analog was, however, found to be an exception. The increase in layer thickness when adding a phosphate group to the threonine is only 35% of that observed for the two other analogs. A detailed infrared examination of the influence of cation coordination to the phosphorylated serine analog using calcium and magnesium reveals structural similarities to those of the inorganic phosphate compound calcium hydroxy apatite. We furthermore discuss the application of these monolayers as soft templates for biomineralization.     

Complex formation in aqueous medium of partially hydrolysed oat cereal proteins with sodium stearoyl-2 lactylate (SSL) lipid surfactant and implications for bile acids activity

Sodium stearoyl-2 lactylate (SSL) lipid surfactant molecules specifically bind partially hydrolysed oat proteins in aqueous medium and significantly enhance the dispersion stability of oat cereal preparations. The proposed complexation is composition dependent and a greater understanding of the role of both oat proteins and lipid surfactant in the effect was gained with data from high performance liquid chromatography (HPLC–UV), viscometry and differential scanning micro calorimetry. The effect of the lipid surfactant on the degree of association is primarily governed by the conformational activity of oat protein molecules related to the extent of protein hydrolysed state, as well as protein unfolded and subsequent aggregated structures. SSL does not dissociate oat proteins into subunits or destroy important hydrophobic contacts already stabilising the protein molecules. Although the exact mode of association is unknown, the present study demonstrates that such interactions occur in a specific manner and suggest selectivity of oat proteins for individual fatty acids. The effect of various amounts of bile acids on SSL–oat protein interaction was also investigated, as a first attempt to investigate the role of lipid surfactant molecules in the known cholesterol-lowering action of oat cereal ingredients and to elucidate favourable conditions by which oat cereal can elicit hypocholesterolemic effects.     

Multilevel Monte Carlo method for parabolic stochastic partial differential equations

We analyze the convergence and complexity of multilevel Monte Carlo discretizations of a class of abstract stochastic, parabolic equations driven by square integrable martingales. We show under low regularity assumptions on the solution that the judicious combination of low order Galerkin discretizations in space and an Euler–Maruyama discretization in time yields mean square convergence of order one in space and of order 1/2 in time to the expected value of the mild solution. The complexity of the multilevel estimator is shown to scale log-linearly with respect to the corresponding work to generate a single path of the solution on the finest mesh, resp. of the corresponding deterministic parabolic problem on the finest mesh.     

In situ Detection of Cobaloxime Intermediates During Photocatalysis Using Hollow-Core Photonic Crystal Fiber Microreactors

Hollow-core photonic crystal fibers (HC-PCFs) provide a novel approach for in situ UV/Vis spectroscopy with enhanced detection sensitivity. Here, we demonstrate that longer optical path lengths than afforded by conventional cuvette-based UV/Vis spectroscopy can be used to detect and identify the Co^I and Co^II states in hydrogen-evolving cobaloxime catalysts, with spectral identification aided by comparison with DFT-simulated spectra. Our findings show that there are two types of signals observed for these molecular catalysts; a transient signal and a steady-state signal, with the former being assigned to the Co^I state and the latter being assigned to the Co^II state. These observations lend support to a unimolecular pathway, rather than a bimolecular pathway, for hydrogen evolution. This study highlights the utility of fiber-based microreactors for understanding these and a much wider range of homogeneous photocatalytic systems in the future.     

Vocal Behavior and Vocal Loading Factors for Preschool Teachers at Work Studied with Binaural DAT Recordings

Preschool teachers are at risk for developing voice problems such as vocal fatigue and vocal nodules. The purpose of this report was to study preschool teachers' voice use during work. Ten healthy female preschool teachers working at daycare centers (DCC) served as subjects. A binaural recording technique was used. Two microphones were placed on both sides of the subject's head, at equal distance from the mouth, and a portable DAT recorder was attached to the subject's waist. Recordings were made of a standard reading passage before work (baseline) and of spontaneous speech during work. The recording technique allowed separate analyses of the level of the background noise, and of the subjects' voice sound pressure level, mean fundamental frequency, and total phonation time. Among the results, mean background noise level for the ten DCCs was 76.1 dBA (range 73.0-78.2), which is more than 20 dB higher than what is recommended where speech communication is important (50-55 dBA). The subjects spoke on an average of 9.1 dB louder (p < 0.0001), and with higher mean fundamental frequency (247 Hz) during work as compared to the baseline (202 Hz) (p < 0.0001). Mean phonation time for the group was 17%, which was considered high. It was concluded that preschool teachers do have a highly vocally demanding profession. Important steps to reduce the vocal loading for this occupation would be to decrease the background noise levels and include pauses so that preschool teachers can rest their voices.     

The role of chromatin conformations in diffusional transport of chromatin-binding proteins: Cartesian lattice simulations

In this paper, a lattice model for the diffusional transport of chromatin-binding particles in the interphase cell nucleus is proposed. Sliding effects are studied in dense networks of chromatin fibers created by three different methods: Randomly distributed, noninterconnected obstacles, a random walk chain model with an attractive step potential, and a self-avoiding random walk chain model with a hard repulsive core and attractive surroundings. By comparing a discrete and continuous version of the random walk chain model, we demonstrate that lattice discretization does not alter the diffusion of chromatin-binding particles. The influence of conformational properties of the fiber network on the particle sliding is investigated in detail while varying occupation volume, sliding probability, chain length, and persistence length. It is observed that adjacency of the monomers, the excluded volume effect incorporated in the self-avoiding random walk model, and the persistence length affect the chromatin-binding particle diffusion. It is demonstrated that sliding particles sense local chain structures. When plotting the diffusion coefficient as a function of the accessible volume for diffusing particles, the data fall onto master curves depending on the persistence length. However, once intersegment transfer is involved, chromatin-binding proteins no longer perceive local chain structures.     

Vanadium oxide compounds with quantum Monte Carlo

Calculations with the diffusion quantum Monte Carlo method are presented for vanadium oxide molecules VO0/+0(n) with n = 1-4 and for V2O5. Atomization and ionization energies are calculated as well as oxygen abstraction energies. The fixed-node approximation is compared for guide functions with orbitals from B3LYP and BP86 calculations and higher accuracy was obtained with the latter orbitals. Additionally, all-electron and pseudopotential calculations are compared for the oxygen atom. The overall accuracy is found to be comparable to CCSD(T) calculations where experimental data is available.     

Efficient bioreduction of bicyclo[2.2.2]octane-2,5-dione and bicyclo[2.2.2]oct-7-ene-2,5-dione by genetically engineered Saccharomyces cerevisiae

A screening of non-conventional yeast species and several Saccharomyces cerevisiae (baker's yeast) strains overexpressing known carbonyl reductases revealed the S. cerevisiae reductase encoded by YMR226c as highly efficient for the reduction of the diketones 1 and 2 to their corresponding hydroxyketones 3-6 (Scheme 1) in excellent enantiomeric excesses. Bioreduction of 1 using the genetically engineered yeast TMB4100, overexpressing YMR226c, resulted in >99% ee for hydroxyketone (+)-4 and 84-98% ee for (-)-3, depending on the degree of conversion. Baker's yeast reduction of diketone 2 resulted in >98% ee for the hydroxyketones (+)-5 and (+)-6. However, TMB4100 led to significantly higher conversion rates (over 40 fold faster) and also a minor improvement of the enantiomeric excesses (>99%).     

An online nano-LC-ESI-FTICR-MS method for comprehensive characterization of endogenous fragments from amyloid β and amyloid precursor protein in human and cat cerebrospinal fluid

Amyloid precursor protein (APP) is the precursor protein to amyloid β (Aβ), the main constituent of senile plaques in Alzheimer's disease (AD). Endogenous Aβ peptides reflect the APP processing, and greater knowledge of different APP degradation pathways is important to understand the mechanism underlying AD pathology. When one analyzes longer Aβ peptides by low-energy collision-induced dissociation tandem mass spectrometry (MS/MS), mainly long b-fragments are observed, limiting the possibility to determine variations such as amino acid variants or post-translational modifications (PTMs) within the N-terminal half of the peptide. However, by using electron capture dissociation (ECD), we obtained a more comprehensive sequence coverage for several APP/Aβ peptide species, thus enabling a deeper characterization of possible variants and PTMs. Abnormal APP/Aβ processing has also been described in the lysosomal storage disease Niemann-Pick type C and the major large animal used for studying this disease is cat. By ECD MS/MS, a substitution of Asp7 → Glu in cat Aβ was identified. Further, sialylated core 1 like O-glycans at Tyr10, recently discovered in human Aβ (a previously unknown glycosylation type), were identified also in cat cerebrospinal fluid (CSF). It is therefore likely that this unusual type of glycosylation is common for (at least) species belonging to the magnorder Boreoeutheria. We here describe a detailed characterization of endogenous APP/Aβ peptide species in CSF by using an online top-down MS-based method.     

Chemical imaging of glucose by CARS microscopy

Glucose is one of the most fundamental molecules within life and bioengineering sciences. Present understanding of its role in cellular and bioengineering processes relies primarily on invasive, large‐scale biochemical analysis, providing no spatial information on glucose pools or fluxes. This work identifies an emerging microscopy technique based on coherent anti‐Stokes Raman scattering (CARS), which fulfills the need of quantitative imaging of glucose at the single‐cell level with submicrometer resolution. No sample preparation with reporter molecules is required, ensuring that the low‐weight metabolite is studied under natural conditions. The potential of CARS microscopy is illustrated by quantitatively mapping glucose fluxes and distributions in a microfluidic bioreactor and in lipid‐bilayer vesicles, the latter as a model for glucose transmembrane transport. Furthermore, the metabolic response to a glucose pulse was monitored in living yeast cells. This study signifies a new era within CARS microscopy for its use of monitoring carbohydrates, in particular glucose which is one of the most abundant molecules in nature. Copyright © 2010 John Wiley & Sons, Ltd.