Protein loaded mesoporous silica spheres as a controlled delivery platform

Background The adsorption of bovine serum albumin (BSA) onto mesoporous silica spheres (MPS) synthesized from silica colloids was studied employing real time in situ measurements. The stabilities of the BSA at different pH values, their isoelectric points and zeta potentials were determined in order to probe the interactions between the protein and the mesoporous silica. Results The pore size of MPS was designed for protein, and this, coupled with an in depth understanding of the physico-chemical characteristics of the protein and MPS has yielded a better binding capacity and delivery profile. The adsorption isotherm at pH 4.2 fitted the Langmuir model and displayed the highest adsorption capacity (71.43 mg mL-1 MPS). Furthermore, the delivery rates of BSA from the MPS under physiological conditions were shown to be dependent on the ionic strength of the buffer and protein loading concentration. Conclusion Economics and scale-up considerations of mesoporous material synthesized via destabilization of colloids by electrolyte indicate the scaleability and commercial viability of this technology as a delivery platform for biopharmaceutical applications.

Rapid production of a plasmid DNA encoding a malaria vaccine candidate via amino-functionalized poly(GMA-co-EDMA) monolith

Malaria is a global health problem; an effective vaccine is urgently needed. Due to the relative poverty and lack of infrastructure in malaria endemic areas, DNA-based vaccines that are stable at ambient temperatures and easy to formulate have great potential. While attention has been focused mainly on antigen selection, vector design and efficacy assessment, the development of a rapid and commercially viable process to manufacture DNA is generally overlooked. We report here a continuous purification technique employing an optimized stationary adsorbent to allow high-vaccine recovery, low-processing time, and, hence, high-productivity. A 40.0 mL monolithic stationary phase was synthesized and functionalized with amino groups from 2-Chloro-N,N- diethylethylamine hydrochloride for anion-exchange isolation of a plasmid DNA (pDNA) that encodes a malaria vaccine candidate, VR1020-PyMSP4/5. Physical characterization of the monolithic polymer showed a macroporous material with a modal pore diameter of 750 nm. The final vaccine product isolated after 3 min elution was homogeneous supercoiled plasmid with gDNA, RNA and protein levels in keeping with clinical regulatory standards. Toxicological studies of the pVR1020-PyMSP4/5 showed a minimum endotoxin level of 0.28 EU/m.g pDNA. This cost-effective technique is cGMP compatible and highly scalable for the production of DNA-based vaccines in commercial quantities, when such vaccines prove to be effective against malaria. © 2008 American Institute of Chemical Engineers.

LacO-LacI interaction in affinity adsorption of plasmid DNA

Current approaches for purifying plasmids from bacterial production systems exploit the physiochemical properties of nucleic acids in non-specific capture systems. In this study, an affinity system for plasmid DNA (pDNA) purification has been developed utilizing the interaction between the lac operon (lacO) sequence contained in the pDNA and a 64mer synthetic peptide representing the DNA-binding domain of the lac repressor protein, LacI. Two plasmids were evaluated, the native pUC19 and pUC19 with dual lacO3/lacOs operators (pUC19lacO3/lacOs), where the lacOs operator is perfectly symmetrical. The DNA-protein affinity interaction was evaluated by surface plasmon resonance using a Biacore system. The affinity capture of DNA in a chromatography system was evaluated using LacI peptide that had been immobilized to Streamline™ adsorbent. The KD-values for double stranded DNA (dsDNA) fragments containing lacO1 and lacO3 and lacOs and lacO3 were 5.7 ± 0.3 × 10 -11 M and 4.1 ± 0.2 × 10-11 M respectively, which compare favorably with literature reports of 5 × 10-10 - 1 × 10-9 M for native laCO1 and 1-1.2 × 10-10 M for lacO1 in a saline buffer. Densitometric analysis of the gel bands from the affinity chromatography run clearly showed a significant preference for capture of the supercoiled fraction from the feed pDNA sample. The results indicate the feasibility of the affinity approach for pDNA capture and purification using native protein-DNA interaction.

Affinity adsorption of plasmid DNA

The development of a protein-mediated dual functional affinity adsorption of plasmid DNA is described in this work. The affinity ligand for the plasmid DNA comprises a fusion protein with glutathione-S-transferase (GST) as the fusion partner with a zinc finger protein. The protein ligand is first bound to the adsorbent by affinity interaction between the GST moeity and gluthathione that is covalently immobilized to the base matrix. The plasmid binding is then enabled via the zinc finger protein and a specific nucleotide sequence inserted into the DNA. At lower loadings, the binding of the DNA onto the Fractogel, Sepharose, and Streamline matrices was 0.0078 ± 0.0013, 0.0095 ± 0.0016, and 0.0080 ± 0.0006 mg, respectively, to 50 μL of adsorbent. At a higher DNA challenge, the corresponding amounts were 0.0179 ± 0.0043, 0.0219 ± 0.0035, and 0.0190 ± 0.0041 mg, respectively. The relatively constant amounts bound to the three adsorbents indicated that the large DNA molecule was unable to utilize the available zinc finger sites that were located in the internal pores and binding was largely a surface adsorption phenomenon. Utilization of the zinc finger binding sites was shown to be highest for the Fractogel adsorbent. The adsorbed material was eluted with reduced glutathione, and the eluted efficiency for the DNA was between 23% and 27%. The protein elution profile appeared to match the adsorption profiles with significantly higher recoveries of bound GST-zinc finger protein.

Fusion of worlds : consumers’ self-narrative in a hybrid physical/digital world

As the Internet becomes deeply embedded into consumers’ daily life, the digital virtual world brings significant influence to consumers’ self and narrative. Prior studies look at consumer self from either from a certain online space or comparing consumers’ physical and digital virtual selves but not the integration of the physical/digital world. This paper aims to explore the meanings of the digital virtual space on consumers’ narrative as a whole (their interests, dreams, or subjectivity). We utilise a postmodern concept of the cyborg to understand the cultural complexity, subjective meanings of, and the extent to which the digital virtual space plays a role in consumers’ self-narrative. We conducted in-depth interviews and gathered three consumer narratives. Our findings indicate that consumers’ narrative contains important fragments from both physical and digital virtual worlds and their physical and digital virtual selves form a feedback loop that strengthen their overall narrative.

The primacy of vital signs – Acute care nurses’ and midwives’ use of physical assessment skills: A cross sectional study

BACKGROUND: Registered nurses and midwives play an essential role in detecting patients at risk of deterioration through ongoing assessment and action in response to changing health status. Yet, evidence suggests that clinical deterioration frequently goes unnoticed in hospitalised patients. While much attention has been paid to early warning and rapid response systems, little research has examined factors related to physical assessment skills. OBJECTIVES: To determine a minimum data set of core skills used during nursing assessment of hospitalised patients and identify nurse and workplace predictors of the use of physical assessment to detect patient deterioration. DESIGN: The study used a single-centre, cross-sectional survey design. SETTING and PARTICIPANTS: The study included 434 registered nurses and midwives (Grades 5-7) involved in clinical care of patients on acute care wards, including medicine, surgery, oncology, mental health and maternity service areas, at a 929-bed tertiary referral teaching hospital in Southeast Queensland, Australia. METHODS: We conducted a hospital-wide survey of registered nurses and midwives using the 133-item Physical Assessment Skills Inventory and the 58-item Barriers to Registered Nurses’ Use of Physical Assessment scale. Median frequency for each physical assessment skill was calculated to determine core skills. To explore predictors of core skill utilisation, backward stepwise general linear modelling was conducted. Means and regression coefficients are reported with 95% confidence intervals. A p value < .05 was considered significant for all analyses. RESULTS: Core skills used by most nurses every time they worked included assessment of temperature, oxygen saturation, blood pressure, breathing effort, skin, wound and mental status. Reliance on others and technology (F = 35.77, p < .001), lack of confidence (F = 5.52, p = .02), work area (F = 3.79, p = .002), and clinical role (F = 44.24, p < .001) were significant predictors of the extent of physical assessment skill use. CONCLUSIONS: The increasing acuity of the acute care patient plausibly warrants more than vital signs assessment; however, our study confirms nurses’ physical assessment core skill set is mainly comprised of vital signs. The focus on these endpoints of deterioration as dictated by early warning and rapid response systems may divert attention from and devalue comprehensive nursing assessment that could detect subtle changes in health status earlier in the patient's hospitalisation.

Ongoing Adaptation as a Feature of Complexity : Further Thoughts and Possible Ideas for Pedagogy in Physical Activity

This chapter takes as its central premise the human capacity to adapt to changing environments. It is an idea that is central to complexity theory but receives only modest attention in relation to learning. To do this we will draw from a range of fields and then consider some recent research in motor control that may extend the discussion in ways not yet considered, but that will build on advances already made within pedagogy and motor control synergies. Recent work in motor control indicates that humans have far greater capacity to adapt to the ‘product space’ than was previously thought, mainly through fast heuristics and on-line corrections. These are changes that can be made in real (movement) time and are facilitated by what are referred to as ‘feed-forward’ mechanisms that take advantage of ultra-fast ways of recognizing the likely outcomes of our movements and using this as a source of feedback. We conclude by discussing some possible ideas for pedagogy within the sport and physical activity domains, the implications of which would require a rethink on how motor skill learning opportunities might best be facilitated.

Seeing it differently: physical education, teacher education and the possibilities of personal construct theory

The celebrated work of Lortie (1975) alerted teacher educators to the extended period of 'apprenticeship' that student teachers have been through before they arrive at teacher education programmes. The subjective implicit theories (Marland, 1992) developed by prospective teachers are shaped by their lifeworld experiences at school and in the case of physical education teachers, their experiences in sport. The biography of physical education teacher education (PETE) students tends to be characterised by ecto-mesomorphic individuals who have been socialised by the rigours of highly competitive sport (Gore, 1990; Macdonald, 1992; Rossi, 1996). We can add to this, the requirements of teacher preparation in physical education which for the most part are dominated by the traditions and rhetoric of the 'natural' bio-physical sciences; largely a legacy of Henry's (1964) work on physical education as an academic discipline, as well as that of Abernathy and Waltz the same year (Abernathy & Waltz, 1964). In the United Kingdom, Curl (1973) further advanced the argument in an attempt to justify human movement as an independent field of study with its own corpus of knowledge. It is little wonder then, that the dominant pedagogical discourse in physical education is, as Tinning (1991) discusses, one of performance pedagogy (see also Hendry, 1986 for an earlier discussion). The knowledge required to support such a discourse could be described as 'official' (Apple, 1993) and it assumes such status by virtue of the power appropriated by and bestowed upon the scientific community in PETE (Macdonald & Tinning, 1995; Sparkes, 1989, 1993). However, there are social reifiers too, and these tend to relate to the social construction of the body (Kirk, 1993; Kirk & Spiller, 1994; Gilroy, 1994) and what Tinning (1985) has termed the Cult of Slenderness. Furthermore the 'slender image' has become a signifier of 'good health'. This is inextricably linked to what might be considered as a health triplex—'exercise = fitness = health' (see Kirk & Colquhoun, 1989; Tinning & Kirk, 1991) which in Australia, underpins curriculum packages such as Daily Physical Education which teachers (often including physical education primary...

Parents’ perception of their children’s participation in physical activity

This paper describes parents' survey and interview responses about their children's participation in physical activity which were collected for the second part of a three phase project funded by the Commonwealth Department of Health and Family Services: Health Advancement Project through the auspices of ACHPER. In the first phase of the project, an extensive data base was compiled on children's participation in physical activity; the second phase investigated parents' and teachers' perceptions of their children's and students' participation in physical activity. The third phase, which is now underway, will use the first two phases to develop a set of resources with which to advocate for policy, programs and educational strategies to serve the needs of young people in Australia most effectively in relation to physical activity.

Children’s physical activity, health and physical education in isolated rural contexts : the views of parent educators in Queensland

This paper describes the views of parent educators of their children’s levels and types of physical activity. The study was conducted at two mini-schools in western Queensland. These are occasion where students who undertake formal education through various Schools Distance Education, come together for a week of educational activity. Parents (mostly mothers) were interviewed using a semi-structured approach. The interview data were then analysed for dominant themes using a constant comparison method. The emergent themes related to nutrition and physical activity. Within the physical activity theme, notions of the great outdoors, work and organised sport skill development also emerged.

Using personal construct theory and narrative methods to facilitate reflexive constructions of teaching physical education

This paper reports on two lengthy studies in Physical education teacher education (PETE) conducted independently but which are epistemologically and methodologically linked. The paper describes how personal construct theory (PCT) and its associated methods provided a means for PETE students to reflexively construct their ideas about teaching physical education over an extended period. Data are drawn from each study in the form of a story of a single participant to indicate how this came about. Furthermore we suggest that PCT might be both a useful research strategy and an effective approach to facilitate professional development in a teacher education setting.

Market segmentation : insights for changing children's physical activity behaviour

Market segmentation has received relatively limited attention in social marketing, particularly within the context of changing children’s physical activity behaviour. This is an important area of investigation given growing concern over childhood obesity globally. The present research aims to extend current understanding of the applicability of market segmentation within this context. The results of a two-step cluster analysis on data from 512 respondents of an online survey show three distinct segments of caregivers, each with unique beliefs about their primary school children walking to/from school. The results demonstrate the validity of employing the process of market segmentation within this social context and provide further insights for targeting the identified segments through tailored social marketing programs.

Intensity and seasonality of physical activity of children during school recess

This study aimed to quantify the intensity of physical activity (PA) of children during school recess (RE), compare the AF gender and seasonal influences. The sample consisted of 30 girls (11.2 ± 1.3 years) and 20 boys (11.3 ± 0.8 years). Heart rate was monitored for three consecutive REs in winter (INV) and spring (PRI) with intensity of the activity being classified as low, moderate and vigorous. Descriptive statistics were used for general data, t test for independent samples for differences between the sexes, paired t test for seasonality. Differences were found between INV and PRI temperatures. Girls had a significant reduction in the AF INV to PRI, which was not observed among boys. The RE represented a small contribution to daily recommendations of AF.