Study of NAP adsorption and assembly on the surface of HOPG

NAP is an octapeptide that has demonstrated a neuroprotective/therapeutic efficacy at very low concentrations in preclinical studies and in a number of clinical trials. Yet little is known about its structural organization at low concentrations. Here, we have employed atomic force microscopy to investigate NAP peptide assembly on graphite in aqueous media at nanomolar concentration. High spatial resolution scans of NAP assemblies reveal their fine structure with clearly resolved single NAP units. This observation leads us to conclude that NAP molecules do not form complex self-assembled structures at nanomolar concentration when adsorbed on graphite surface

Real-time measurement of the intracellular pH of yeast cells during glucose metabolism using ratiometric fluorescent nanosensors

Intracellular pH is a key parameter that influences many biochemical and metabolic pathways that can also be used as an indirect marker to monitor metabolic and intracellular processes. Herein, we utilise ratiometric fluorescent pH-sensitive nanosensors with an extended dynamic pH range to measure the intracellular pH of yeast (Saccharomyces cerevisiae) during glucose metabolism in real-time. Ratiometric fluorescent pH-sensitive nanosensors consisting of a polyacrylamide nanoparticle matrix covalently linked to two pH-sensitive fluorophores, Oregon green (OG) and 5(6)carboxyfluorescein (FAM), and a reference pH-insensitive fluorophore, 5(6)carboxytetramethylrhodamine (TAMRA), were synthesised. Nanosensors were functionalised with acrylamidopropyltrimethyl ammonium hydrochloride (ACTA) to confer a positive charge to the nanoparticle surfaces that facilitated nanosensor delivery to yeast cells, negating the need to use stress inducing techniques. The results showed that under glucose-starved conditions the intracellular pH of yeast population (n ≈ 200) was 4.67 ± 0.15. Upon addition of D-(+)-glucose (10 mM), this pH value decreased to pH 3.86 ± 0.13 over a period of 10 minutes followed by a gradual rise to a maximal pH of 5.21 ± 0.26, 25 minutes after glucose addition. 45 minutes after the addition of glucose, the intracellular pH of yeast cells returned to that of the glucose starved conditions. This study advances our understanding of the interplay between glucose metabolism and pH regulation in yeast cells, and indicates that the intracellular pH homestasis in yeast is highly regulated and demonstrates the utility of nanosensors for real-time intracellular pH measurements

Disruption of diphenylalanine assembly by a Boc-modified variant

Peptide-based biomaterials are key to the future of diagnostics and therapy, promoting applications such as tissue scaffolds and drug delivery vehicles. To realise the full potential of the peptide systems, control and optimisation of material properties are essential. Here we invesigated the co-assembly of the minimal amyloid motif peptide, diphenylalanine (FF), and its tert-butoxycarbonyl (Boc)-modified derivative. Using Atomic Force Microscopy, we demonstrated that the co-assembled fibers are less rigid and show a curvier morphology. We propose that the Boc-modification of FF disrupts the hydrogen bond packing of adjacent N-termini, as supported by Fourier transform infrared and fluorescence spectroscopic data. Such rationally modified co-assemblies offer chemical functionality for after-assembly modification and controllable surface properties for tissue engineering scaffolds, along with tunable morphological vs. mechanical properties

Enhanced distance-dependent fluorescence quenching using size tuneable core shell silica nanoparticles

Silica nanoparticles (SNPs) have been used as favoured platforms for sensor, drug delivery and biological imaging applications, due to their ease of synthesis, size-control and bespoke physico-chemical properties. In this study, we have developed a protocol for the synthesis of size-tuneable SNPs, with diameters ranging from 20 nm to 500 nm, through the optimisation of experimental components required for nanoparticle synthesis. This protocol was also used to prepare fluorescent SNPs, via covalent linkages of fluorophores, to the nanoparticle matrix using 3-aminopropyltriethoxysilane (APTES). This enabled the fabrication of ratiometric, fluorescent, pH-sensitive nanosensors (75 nm diameter) composed SNPs covalently linked to two pH-sensitive fluorescent dyes Oregon Green (OG) and 5(6)-carboxyfluorescein (FAM) and a reference fluorescent dye 5-(6)-carboxytetramethylrhodamine (TAMRA), extending the dynamic range of measurement from pH 3.5 to 7.5. In addition, size-tuneable, core-shell SNPs, covalently linked to a fluorescent TAMRA core were synthesised to investigate distance-dependant fluorescence quenching between TAMRA and black hole quencher 2 (BHQ2®) using nanometre-sized silica shells as physical spacers. The results showed a significant fluorescence quenching could be observed over greater distances than that reported for the classical distance-dependent molecular fluorescence quenching techniques, e.g. the Förster (fluorescence) resonance energy transfer (FRET). The methods and protocols we have detailed in this manuscript will provide the basis for the reproducible production of size tunable SNPs, which will find broad utility in the development of sensors for biological applications

O futuro da profissão de auditoria

Technology is evolving at an unbridled pace, some sectors are not failing to keep up with this evolution. It is estimated that the information processed worldwide in the last 2 years represents 90% of the total information ever created. Software is replacing professionals all over the world, mainly in areas such as accounting, routine data processing tasks are performed without human input and automatically. As the audit profession is highly dependent on data analysis, it is important to understand the impact that this evolution will have on the auditor's life. This dissertation aims to describe the audit profession in this context of global technological evolution. The literature review made it possible to fit into important theoretical concepts and understand how these concepts translate into the audit profession. With the empirical study carried out, the objective is to demonstrate the impacts of these new technologies on the auditor's life. The empirical study was supported by a research with a quantitative analysis, through the use of an inquiry. Through this, it was possible to conclude that the new technologies, despite presenting new risks for the external audit, also present themselves as an opportunity to develop works with more quality and efficiency.A tecnologia está a evoluir a um ritmo desenfreado, alguns setores não estão a conseguir acompanhar esta evolução. Estima-se que a informação processada mundialmente nos últimos 2 anos representa 90% do total de informação alguma vez criada. Softwares estão a substituir profissionais pelo mundo inteiro, principalmente em áreas como a contabilidade, tarefas rotineiras de processamento de dados são desempenhadas sem contributo humano e de forma automática. Sendo a profissão de auditoria altamente dependente de análise de dados é importante perceber o impacto que esta evolução terá na vida do auditor. Esta dissertação tem como finalidade descrever a profissão de auditoria neste contexto de evolução tecnológica global. A revisão de literatura efetuada permitiu enquadrar em importantes conceitos teóricos e perceber como esses conceitos se traduzem para a profissão de auditoria. Com o estudo empírico realizado, procurou-se demonstrar os impactos destas novas tecnologias na vida do auditor. O estudo empírico suportou-se numa pesquisa com uma análise quantitativa, através da utilização de um inquérito. Através deste, foi possível concluir que as novas tecnologias apesar de apresentarem novos riscos para a auditoria externa, também se apresentam como uma oportunidade para desenvolver trabalhos com mais qualidade e eficiência

The two-dimensional random-bond Ising model, free fermions and the network model

We develop a recently-proposed mapping of the two-dimensional Ising model with random exchange (RBIM), via the transfer matrix, to a network model for a disordered system of non-interacting fermions. The RBIM transforms in this way to a localisation problem belonging to one of a set of non-standard symmetry classes, known as class D; the transition between paramagnet and ferromagnet is equivalent to a delocalisation transition between an insulator and a quantum Hall conductor. We establish the mapping as an exact and efficient tool for numerical analysis: using it, the computational effort required to study a system of width $M$ is proportional to $M^{3}$, and not exponential in $M$ as with conventional algorithms. We show how the approach may be used to calculate for the RBIM: the free energy; typical correlation lengths in quasi-one dimension for both the spin and the disorder operators; even powers of spin-spin correlation functions and their disorder-averages. We examine in detail the square-lattice, nearest-neighbour $\pm J$ RBIM, in which bonds are independently antiferromagnetic with probability $p$, and ferromagnetic with probability $1-p$. Studying temperatures $T\geq 0.4J$, we obtain precise coordinates in the $p-T$ plane for points on the phase boundary between ferromagnet and paramagnet, and for the multicritical (Nishimori) point. We demonstrate scaling flow towards the pure Ising fixed point at small $p$, and determine critical exponents at the multicritical point.Comment: 20 pages, 25 figures, figures correcte

Instability of the rhodium magnetic moment as origin of the metamagnetic phase transition in alpha-FeRh

Based on ab initio total energy calculations we show that two magnetic states of rhodium atoms together with competing ferromagnetic and antiferromagnetic exchange interactions are responsible for a temperature induced metamagnetic phase transition, which experimentally is observed for stoichiometric alpha-FeRh. A first-principle spin-based model allows to reproduce this first-order metamagnetic transition by means of Monte Carlo simulations. Further inclusion of spacial variation of exchange parameters leads to a realistic description of the experimental magneto-volume effects in alpha-FeRh.Comment: 10 pages, 13 figures, accepted for publication in Phys. Rev.

Isometric Sliced Inverse Regression for Nonlinear Manifolds Learning

[[abstract]]Sliced inverse regression (SIR) was developed to find effective linear dimension-reduction directions for exploring the intrinsic structure of the high-dimensional data. In this study, we present isometric SIR for nonlinear dimension reduction, which is a hybrid of the SIR method using the geodesic distance approximation. First, the proposed method computes the isometric distance between data points; the resulting distance matrix is then sliced according to K-means clustering results, and the classical SIR algorithm is applied. We show that the isometric SIR (ISOSIR) can reveal the geometric structure of a nonlinear manifold dataset (e.g., the Swiss roll). We report and discuss this novel method in comparison to several existing dimension-reduction techniques for data visualization and classification problems. The results show that ISOSIR is a promising nonlinear feature extractor for classification applications.[[incitationindex]]SCI[[booktype]]紙本[[booktype]]電子