Elucidating the Influence of Electrical Potentials on the Formation of Charged Oligopeptide Self-Assembled Monolayers on Gold

Self-assembled monolayers (SAMs) based on oligopeptides have garnered immense interest for a wide variety of innovative biomedical and electronic applications. However, to exploit their full potential, it is necessary to understand and control the surface chemistry of oligopeptides. Herein, we report on how different electrical potentials affect the adsorption kinetics, stability and surface coverage of charged oligopeptide SAMs on gold surfaces. Kinetic analysis using electrochemical surface plasmon resonance (e-SPR) reveals a slower oligopeptide adsorption rate at more positive or negative electrical potentials. Additional analysis of the potential-assisted formed SAMs by X-ray photoelectron spectroscopy demonstrates that an applied electrical potential has minimal effect on the packing density. These findings not only reveal that charged oligopeptides exhibit a distinct potential-assisted assembly behaviour but that an electrical potential offers another degree of freedom in controlling their adsorption rate.     

3-O-Sulfation of Heparan Sulfate Enhances Tau Interaction and Cellular Uptake

Prion-like transcellular spreading of tau in Alzheimer's Disease (AD) is mediated by tau binding to cell surface heparan sulfate (HS). However, the structural determinants for tau–HS interaction are not well understood. Microarray and SPR assays of structurally defined HS oligosaccharides show that a rare 3-O-sulfation (3-O-S) of HS significantly enhances tau binding. In Hs3st1^−/− (HS 3-O-sulfotransferase-1 knockout) cells, reduced 3-O-S levels of HS diminished both cell surface binding and internalization of tau. In a cell culture, the addition of a 3-O-S HS 12-mer reduced both tau cell surface binding and cellular uptake. NMR titrations mapped 3-O-S binding sites to the microtubule binding repeat 2 (R2) and proline-rich region 2 (PRR2) of tau. Tau is only the seventh protein currently known to recognize HS 3-O-sulfation. Our work demonstrates that this rare 3-O-sulfation enhances tau–HS binding and likely the transcellular spread of tau, providing a novel target for disease-modifying treatment of AD and other tauopathies.     

The First Turbulence and First Fossil Turbulence

A model is proposed connecting turbulence, fossil turbulence and the big-bang origin of the universe. While details are incomplete, the model is consistent with our knowledge of these processes and is supported by observations. Turbulence arises in a hot big-bang quantum gravitational dynamics scenario at Planck scales. Chaotic, eddy-like motions produce an exothermic Planck particle cascade from 10^?35 m at 10³² K to 10⁸ larger, 10⁴ cooler, quark-gluon scales. A Planck-Kerr instability gives high Reynolds number (Re ～ 10⁶) turbulent combustion, space-time-energy-entropy and turbulent mixing. Batchelor-Obukhov-Corrsin turbulent-temperature fluctuations are preserved as the first fossil turbulence by inflation stretching the patterns beyond the horizon ct of causal connection faster than light speed c in time t～ 10^?33 sec. Fossil big-bang temperature turbulence reenters the horizon and imprints nucleosynthesis of H-He densities that seed fragmentation by gravity at 10¹² s in the low Reynolds number plasma before its transition to gas at t～ 10¹³ s and T～ 3000 K. Multiscaling coefficients of the cosmic microwave background (CMB) temperature anisotropies closely match those for high Reynolds number turbulence, Bershadskii, A. and Sreenivasan, K.R., Phys. Lett. A 299 (2002) 149-152; Bershadskii, A. and Sreenivasan, K.R., Phys. Lett. A 319 (2003) 21-23. CMB spectra support the interpretation that big-bang turbulence fossils triggered fragmentation of the viscous plasma at supercluster to galaxy mass scales from 10⁴⁶ to 10⁴² kg, Gibson, C.H., Appl. Mech. Rev. 49 (5) (1996) 299-315; Gibson, C.H., J. Fluids Eng. 122 (2000) 830-835; Gibson, C.H., Combust. Sci. Technol. (2004, to be published).     

Turbulent boundary layer on a mildly curved convex surface

Mean flow and turbulence measurements have been made in a boundary layer which grows first on a flat' wall and then on a convex wall of radius of curvature approximately 100 times the boundary layer thickness. The turbulence data include profiles of the four non-zero components of the Reynolds stress tensor and three triple velocity products obtained at five stream-wise positions. A number of measurements were also made for comparison in the boundary layer on a flat wall under the same conditions. The effects of convex curvature are to reduce turbulent intensities, shear stress and wall friction by approximately 10% of their plane flow values; the triple velocity products are halved in the curved layer. The measurements supplement the small quantity of previously published data available for testing mathematical models of turbulence. The results show the same general trends that have been observed in earlier investigations but there are significant differences in detail, notably in respect of levels of the normal stresses.     

Strain strengthening effects in Witwatersrand quartzite

A series of uniaxial compression specimens were tested over a range of applied ram displacement rates of 8.9 × 10⁻⁴ to 8.9 mm/sec to elucidate the effects of loading rate on the uniaxial compressive fracture stress of Witwatersrand quartzite. It was demonstrated that even within standard loading rate ranges, considerable scatter in the fracture strength (under uniaxial compression) existed in this particular quartzite rock. Nevertheless, a definite trend of increasing fracture resistance with increasing monotonic loading rate was evident inasmuch that increasing the loading rate (strain rate) by four orders of magnitude increase the fracture strength by almost 2.8 times. Prior fatigue loading also produced a significant strain strengthening as the uniaxial compressive fracture stress tended to increase in a sigmoidal fashion with increasing number of fatigue cycles prior to testing. Indeed, the fracture strength of quartzite was almost doubled in value after 10⁻ cycles. Plane strain fracture toughness tests utilising three point bend specimens were conducted and an average of K_lc = 1.7 MPa√m was realized. In both the uniaxial compression tests and the fracture toughness tests, failure occurred by crack extension predominantly by a transgranular flat cleavage-like mode through pure quartzite (silica) regions. However, crack extension was also observed to occur in an intergranular “ductile-like” mode through areas associated with inclusions prevalent in the quartzite.     

Comparison of RAFT‐derived poly(vinylpyrrolidone) verses poly(oligoethyleneglycol methacrylate) for the stabilization of glycosylated gold nanoparticles

Carbohydrates dictate many biological processes including infection by pathogens. Glycosylated polymers and nanomaterials which have increased affinity due to the cluster glycoside effect, are therefore useful tools to probe function, but also as prophylactic therapies or diagnostic tools. Here, the effect of polymer structure on the coating of gold nanoparticles is studied in the context of grafting density, buffer stability, and in a lectin binding assay. RAFT polymerization is used to generate poly(oligoethyleneglycol methacrylates) and poly(N‐vinylpyrrolidones) with a thiol end‐group for subsequent immobilization onto the gold. It is observed that poly(oligoethylene glycol methacrylates), despite being widely used particle coatings, lead to low grafting densities which in turn resulted in lower stability in biological buffers. A depression of the cloud point upon nanoparticle immobilization is also seen, which might compromise performance. In comparison poly(vinylpyrrolidones) resulted in stable particles with higher grafting densities due to the compact size of each monomer unit. The higher grafting density also enabled an increase in the number of carbohydrates which can be installed per nanoparticle at the chain ends, and gave increased binding in a lectin recognition assay. These results will guide the development of new nanoparticle biosensors with enhanced specificity, affinity, and stability. © 2016 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2016 , 55, 1200–1208     

Radiolabelling of TiO<Subscript>2</Subscript> nanoparticles for radiotracer studies

Industrially manufactured titanium dioxide nanoparticles have been successfully radiolabelled with ⁴⁸V by irradiation with a cyclotron-generated proton beam. Centrifugation tests showed that the ⁴⁸V radiolabels were stably bound within the nanoparticle structure in an aqueous medium, while X-ray diffraction indicated that no major structural modifications to the nanoparticles resulted from the proton irradiation. In vitro tests of the uptake of cold and radiolabelled nanoparticles using the human cell line Calu-3 showed no significant difference in the uptake between both batches of nanoparticles. The uptake was quantified by Inductively Coupled Plasma Mass Spectrometry and high resolution γ-ray spectrometry for cold and radiolabelled nanoparticles, respectively. These preliminary results indicate that alterations to the nanoparticles’ properties introduced by proton bombardment can be controlled to a sufficient extent that their further use as radiotracers for biological investigations can be envisaged and elaborated.     

Diversity oriented synthesis: substitution at C5 in unreactive pyrimidines by Claisen rearrangement and reactivity in nucleophilic substitution at C2 and C4 in pteridines and pyrido[2,3-d]pyrimidines

Diversity oriented synthesis of fused pyrimidines leads to scaffolds with many biological activities. In the case of the preparation of pyrido[2,3-d]pyrimidines from 2-alkylthiopyrimidines, the formation of a new carbon-carbon bond at C5 is required, a reaction, that is, very limited in scope. However Claisen type rearrangement of simple 4-allylic ethers affords C5 substituted pyrimidines readily; in cases with an ester substituent, rearrangement occurs at room temperature. Subsequent cyclisation to afford 6-methylpyrido[2,3-d]pyrimidin-7(8H)-ones was achieved in high yield. Using allylic ethers derived from 3-chloromethyl-4-arylbut-3-en-2-ones as substrates, a new titanium[IV]chloride catalysed reaction affording 6-arylmethyl-7-methylpyrido[2,3-d]pyrimidines was discovered. In contrast, 2-alkylthiopteridines are readily available. In both cases, substitution at C2 and C4 to generate diversity has been carried out and the reactivity compared; yields of substitution products were generally higher with pteridine substrates. In biological assays unexpected hits were found for activity against the Gram positive bacterium, Nocardia farcinia, and against the parasite Trypanosoma brucei brucei, illustrating the value of diversity oriented synthesis in the discovery of biologically active compounds.     

Generation and characterization of stable, highly concentrated titanium dioxide nanoparticle aerosols for rodent inhalation studies

The intensive use of nano-sized titanium dioxide (TiO₂) particles in many different applications necessitates studies on their risk assessment as there are still open questions on their safe handling and utilization. For reliable risk assessment, the interaction of TiO₂ nanoparticles (NP) with biological systems ideally needs to be investigated using physico-chemically uniform and well-characterized NP. In this article, we describe the reproducible production of TiO₂ NP aerosols using spark ignition technology. Because currently no data are available on inhaled NP in the 10?C50 nm diameter range, the emphasis was to generate NP as small as 20 nm for inhalation studies in rodents. For anticipated in vivo dosimetry analyses, TiO₂ NP were radiolabeled with ⁴⁸V by proton irradiation of the titanium electrodes of the spark generator. The dissolution rate of the ⁴⁸V label was about 1% within the first day. The highly concentrated, polydisperse TiO₂ NP aerosol (3?C6 × 10⁶ cm^?3) proved to be constant over several hours in terms of its count median mobility diameter, its geometric standard deviation, and number concentration. Extensive characterization of NP chemical composition, physical structure, morphology, and specific surface area was performed. The originally generated amorphous TiO₂ NP were converted into crystalline anatase TiO₂ NP by thermal annealing at 950 °C. Both crystalline and amorphous 20-nm TiO₂ NP were chain agglomerated/aggregated, consisting of primary particles in the range of 5 nm. Disintegration of the deposited TiO₂ NP in lung tissue was not detectable within 24 h.     

3‐O‐Sulfation of Heparan Sulfate Enhances Tau Interaction and Cellular Uptake

Prion‐like transcellular spreading of tau in Alzheimer's Disease (AD) is mediated by tau binding to cell surface heparan sulfate (HS). However, the structural determinants for tau–HS interaction are not well understood. Microarray and SPR assays of structurally defined HS oligosaccharides show that a rare 3‐O‐sulfation (3‐O‐S) of HS significantly enhances tau binding. In Hs3st1^?/? (HS 3‐O‐sulfotransferase‐1 knockout) cells, reduced 3‐O‐S levels of HS diminished both cell surface binding and internalization of tau. In a cell culture, the addition of a 3‐O‐S HS 12‐mer reduced both tau cell surface binding and cellular uptake. NMR titrations mapped 3‐O‐S binding sites to the microtubule binding repeat 2 (R2) and proline‐rich region 2 (PRR2) of tau. Tau is only the seventh protein currently known to recognize HS 3‐O‐sulfation. Our work demonstrates that this rare 3‐O‐sulfation enhances tau–HS binding and likely the transcellular spread of tau, providing a novel target for disease‐modifying treatment of AD and other tauopathies.