Immobilisation of Bio-molecules on Magnetisable Solid Supports for Applications in Bio-catalysts and Bio-sensors

A series of core and core-shell nanoparticles with superparamagnetic properties were synthesised and surface functionalised using three different amino-silanes by a chemical conjugation method. The functionalised nanoparticles were characterised and further modified by chemical conjugation with two different classes of bio-molecules; (i) enzymes and (ii) single stranded DNA primers. The resultant nanoparticles (nano-bio conjugates) were used for applications in (i) enzyme catalysis and (ii) bio-separation / bio-sensing. Magnetite and amorphous silica-coated core-shell nanoparticles were synthesised on both small (5 g) and large (20 g) scales and were characterised using transmission electron microscopy (TEM), X-ray diffraction (XRD), Brunauer-Emmett-Teller (BET) surface area measurement and vibrating sample magnetometry (VSM). Silica-coated core-shell nanoparticles were functionalised by silanisation with three different aminosilanes [3-aminopropyl tri-ethoxysilane (APTS), 3-aminopropyl di-ethoxymethylsilane (APDS) and 3-aminopropyl mono-ethoxydimethylsilane (APMS)] and two different methods: water (classical method) or a Tri-phasic Reverse Emulsion (TPRE) using toluene and a surfactant (Triton X-100). It was observed that the materials prepared using the TPRE method produced higher surface amine density values on average. The first application involved bio-catalysis where lipases [Pseudomonas Fluorescens lipase (PFL) and Candida Rugosa lipase (CRL)] were chemically conjugated (covalently linked) via glutaraldehyde-modification onto the amino-functionalised nanoparticles for applications such as: (i) hydrolysis of p-nitrophenyl palmitate to produce palmitic acid and p-nitrophenol (model reaction), (ii) transesterification of ethyl butyrate with n-butanol to produce butyl butyrate and (iii) partial and selective hydrolysis of cis-3,5-diacetoxy-1-cyclopentene to produce pharmaceutically important and expensive chiral intermediate molecules. Various reaction parameters such as (a) water concentration in a bi-phasic solvent mixture and (b) temperature were investigated to determine the optimum conditions. All reactions were carried out using free lipases and the physically adsorbed lipases in order to compare the performance with chemically conjugated nano-biomaterials. It was observed from the bio-catalytic reaction (i) that the conversion values given by lipase-immobilised materials were comparable to those given by free lipases with the added advantage of being re-usable for further catalytic cycles. PFL-immobilised nanoparticles were shown to be more effective catalysts than CRL-immobilised materials. In the bio- catalytic reaction (ii), Lipase-immobilised materials were shown to exhibit reasonable conversion values (maximum 53%) along with easy separability by one-step magnetic separation from the reaction mixture and re-usability. Finally, in the bio-catalytic reaction (iii), lipase-immobilised materials were shown to give lower total conversion values compared to free enzymes, but a higher proportion of desired products [(1S,4R)-cis-4-acetoxy-2-cyclopenten-1-ol and (1R,4S)-cis-4-acetoxy-2-cyclopenten-1-ol]. PFL (both free and immobilised) materials were shown to give higher conversion and enantioselectivity towards the desired (1S,4R)-enantiomer (93-100% ee) than CRL materials (30-40% ee). The second application involved bio-separation and bio-sensing where 5ʹ-NH2-modified oligonucleotide sequences specific to either Listeria Monocytogenes (LM) or Escherichia Coli (EC) were immobilised onto the surface of glutaraldehyde modified nanoparticles to assess the specific capture and enhance the sensitivity of detection of pathogenic bacterial DNAs from food samples. Firstly, the oligonucleotide-grafted nanoparticles were used in a hybrid capture assay (model assay) at UCLan using specific single stranded DNA primers of our interest followed by the application in real food samples at Q-Bioanalytic GmbH, Germany. Capture of the complementary sequences was reasonably high (48-70% for LM-specific materials and 48-55% for EC-specific materials) when calculated as a molar ratio of conjugated oligonucleotides to complementary oligonucleotides captured. Specific capture was determined to be 33-52% for LM-specific oligonucleotide-grafted nano-materials and 59-60% for EC-specific oligonucleotide-grafted nano-materials. Dehybridisation of captured sequences was shown to be efficient for all oligonucleotide-grafted materials (72-97% for LM-specific materials and 86-87% for EC-specific materials), indicating that the materials were ready for real applications using food matrices at Q-Bioanalytic GmbH, Germany. Nucleic acid DNA was extracted from a real food sample inoculated with either LM or EC and the extracted DNA was used for specific capture using the oligonucleotide-grafted materials tested at UCLan. Dehybridised oligonucleotides were amplified and analysed using quantitative real-time PCR (qPCR). The results showed that using a one-step hybrid capture assay, LM-specific oligonucleotide-grafted materials were successful at detecting LM from an undiluted solution of LM only and from a 1:1 mixture of LM and EC. Using a two-step assay where the forward and reverse oligonucleotide-grafted materials were applied for capture separately, only EC-specific materials were successful for the detection of EC from an undiluted solution, and also from a 1:1 mixture of LM and EC

Sheet forming simulation for AHSS components in the automotive industry

The trend in the automotive industry towards new advanced high strength steels (AHSS), combined with the ongoing reduction in program lead times have increased the need to get tool designs right, first time. Despite the fact that the technology used by sheet metal stamping companies to design and manufacture tooling is advancing steadily, finding optimal process parameters and tool geometries remains a challenge. Consequently, there has been a transition from designs based largely on trial and error techniques and the experience of the stamping engineer, to the increased use of virtual manufacturing and finite element (FE) simulation predictions as an indispensable tool in the design process. This work investigates the accuracy of FE techniques in predicting the forming behavior of AHSS grades, such as TRIP and dual phase, as compared to more commonly used conventional steel grades. Three different methods of simulation, one-step, implicit and explicit techniques, were used to model the forming process for an automotive part. Results were correlated with experimental strain and thickness measurements of manufactured components from the production line.<br /

Practical, appropriate, empirically-validated guidelines for designing educational games

There has recently been a great deal of interest in the potential of computer games to function as innovative educational tools. However, there is very little evidence of games fulfilling that potential. Indeed, the process of merging the disparate goals of education and games design appears problematic, and there are currently no practical guidelines for how to do so in a coherent manner. In this paper, we describe the successful, empirically validated teaching methods developed by behavioural psychologists and point out how they are uniquely suited to take advantage of the benefits that games offer to education. We conclude by proposing some practical steps for designing educational games, based on the techniques of Applied Behaviour Analysis. It is intended that this paper can both focus educational games designers on the features of games that are genuinely useful for education, and also introduce a successful form of teaching that this audience may not yet be familiar with

On a new conformal functional for simplicial surfaces

We introduce a smooth quadratic conformal functional and its weighted version $$W_2=\sum_e \beta^2(e)\quad W_{2,w}=\sum_e (n_i+n_j)\beta^2(e),$$ where $\beta(e)$ is the extrinsic intersection angle of the circumcircles of the triangles of the mesh sharing the edge $e=(ij)$ and $n_i$ is the valence of vertex $i$. Besides minimizing the squared local conformal discrete Willmore energy $W$ this functional also minimizes local differences of the angles $\beta$. We investigate the minimizers of this functionals for simplicial spheres and simplicial surfaces of nontrivial topology. Several remarkable facts are observed. In particular for most of randomly generated simplicial polyhedra the minimizers of $W_2$ and $W_{2,w}$ are inscribed polyhedra. We demonstrate also some applications in geometry processing, for example, a conformal deformation of surfaces to the round sphere. A partial theoretical explanation through quadratic optimization theory of some observed phenomena is presented.Comment: 14 pages, 8 figures, to appear in the proceedings of "Curves and Surfaces, 8th International Conference", June 201

Psychological distress and quality of life in asymptomatic adults following provision of imaging results for prevention of cardiovascular disease events: A scoping review

Aims Psychological distress and changes in health-related quality of life (HRQoL) may occur after screening for disease. Reporting outcomes related to potential benefits and harms of screening is a key recommendation in the guidelines for reporting high-quality trials or interventions. However, no reviews have directly investigated outcomes related to psychological distress and/or changes in HRQoL following imaging assessment of cardiovascular risk and communication of cardiovascular findings to asymptomatic adults. A scoping review was conducted to map research on psychological distress and/or HRQoL following screening. Methods and results Six electronic databases (MEDLINE, PsychINFO, Social Work Abstracts, Psychology and Behavioural Sciences Collection, CINAHL, and EMBASE) were searched for articles that assessed psychological distress and/or HRQoL following screening. Two investigators independently screened titles and abstracts for all records retrieved using predefined criteria. Studies were conducted among active smokers, military personnel, athletes, post-menopausal women, and high-risk individuals. Seven constructs related to psychological distress and HRQoL appeared across 11 articles (randomized controlled trials, n = 4 and non-randomized studies, n = 7). Worry, depression, perceived stress, anxiety, and quality of life were most prominent. Multiple-item measures of psychological distress (e.g. Taylor Anxiety Score and Beck Depression Inventory) were used in 5/9 (56%) studies. Key findings on psychological distress and/or changes in HRQoL following screening were mixed. Conclusions Findings support the need for multiple-item measures with better psychometric properties to examine the psychological responses to screening results in future studies. Strategies to support individuals during and following vascular screening to maximise potential benefits of screening and minimize harms are discussed

Psychological distress and quality of life in asymptomatic adults following provision of imaging results for prevention of cardiovascular disease events: a scoping review

Aims: Psychological distress and changes in health-related quality of life (HRQoL) may occur after screening for disease. Reporting outcomes related to potential benefits and harms of screening is a key recommendation in the guidelines for reporting high-quality trials or interventions. However, no reviews have directly investigated outcomes related to psychological distress and/or changes in HRQoL following imaging assessment of cardiovascular risk and communication of cardiovascular findings to asymptomatic adults. A scoping review was conducted to map research on psychological distress and/or HRQoL following screening. Methods and results: Six electronic databases (MEDLINE, PsychINFO, Social Work Abstracts, Psychology and Behavioural Sciences Collection, CINAHL, and EMBASE) were searched for articles that assessed psychological distress and/or HRQoL following screening. Two investigators independently screened titles and abstracts for all records retrieved using predefined criteria. Studies were conducted among active smokers, military personnel, athletes, post-menopausal women, and high-risk individuals. Seven constructs related to psychological distress and HRQoL appeared across 11 articles (randomized controlled trials, n = 4 and non-randomized studies, n = 7). Worry, depression, perceived stress, anxiety, and quality of life were most prominent. Multiple-item measures of psychological distress (e.g. Taylor Anxiety Score and Beck Depression Inventory) were used in 5/9 (56%) studies. Key findings on psychological distress and/or changes in HRQoL following screening were mixed. Conclusions: Findings support the need for multiple-item measures with better psychometric properties to examine the psychological responses to screening results in future studies. Strategies to support individuals during and following vascular screening to maximise potential benefits of screening and minimize harms are discussed

Impact of vascular screening interventions on perceived threat, efficacy beliefs and behavioural intentions: A systematic narrative review

Health-related behaviours contribute to the global burden of cardiovascular disease (CVD). Cardiovascular imaging can be used to screen asymptomatic individuals for increased risk of CVD to enable earlier interventions to promote health-related behaviours to prevent or reduce CVD risk. Some theories of behaviour and behaviour change assume that engagement in a given behaviour is a function of individual threat appraisals, beliefs regarding the performance of behaviour, self-efficacy for performing the desired behaviour and/or dispositions to act (e.g. behavioural intentions). To date, little is known about the impact of cardiovascular imaging interventions on these constructs. This article summarises evidence related to perceived threat, efficacy beliefs, and behavioural intentions after CVD screening. We identified 10 studies (2 RCTs and 8 non-randomised studies, n = 2498) through a combination of screening citations from published systematic reviews and meta-analyses and searching electronic databases. Of these, 7 measured behavioural intentions and perceived susceptibility and 3 measured efficacy beliefs. Findings showed largely encouraging effects of screening interventions on bolstering self-efficacy beliefs and strengthening behavioural intentions. Imaging results that suggest the presence of coronary or carotid artery disease also increased perceived susceptibility to CVD. However, the review also identified some gaps in the literature, such as a lack of guiding theoretical frameworks and assessments of critical determinants of health-related behaviours. By carefully considering the key issues highlighted in this review, we can make significant strides towards reducing CVD risks and improving population health