Three-dimensional fluid pressure mapping in porous media using magnetic resonance imaging with gas-filled liposomes

This paper presents and demonstrates a method for using magnetic resonance imaging to measure local pressure of a fluid saturating a porous medium. The method is tested both in a static system of packed silica gel and in saturated sintered glass cylinders experiencing fluid flow. The fluid used contains 3% gas in the form of 3-μm average diameter gas filled 1,2-distearoyl-sn-glycero-3-phosphocholine (C18:0, MW: 790.16) liposomes suspended in 5% glycerol and 0.5% Methyl cellulose with water. Preliminary studies at 2.35 T demonstrate relative magnetic resonance signal changes of 20% per bar in bulk fluid for an echo time TE=40 ms, and 6-10% in consolidated porous media for TE=10 ms, over the range 0.8-1.8 bar for a spatial resolution of 0.1 mm3 and a temporal resolution of 30 s. The stability of this solution with relation to applied pressure and methods for improving sensitivity are discussed. © 2007 Elsevier Inc. All rights reserved.

An initial evaluation of gellan gum as a material for tissue engineering applications

Alpha-modified minimum essential medium (αMEM) has been found to cross-link a 1% gellan gum solution, resulting in the formation of a self-supporting hydrogel in 1:1 and 5:1 ratios of polysaccharide: αMEM. Rheological data from temperature sweeps confirm that in addition to orders of magnitude differences in G' between 1% gellan and 1% gellan with αMEM, there is also a 20°C increase in the temperature at which the onset of gelation takes place when αMEM is present. Frequency sweeps confirm the formation of a true gel; mechanical spectra for mixtures of gellan and αMEM clearly demonstrate G' to be independent of frequency. It is possible to immobilize cells within a three-dimensional (3D) gellan matrix that remain viable for up to 21 days in culture by adding a suspension of rat bone marrow cells (rBMC) in αMEM to 1% gellan solution. This extremely simple approach to cell immobilization within 3D constructs, made possible by the fact that gellan solutions cross-link in the presence of millimolar concentrations of cations, poses a very low risk to a cell population immobilized within a gellan matrix and thus indicates the potential of gellan for use as a tissue engineering scaffold. © 2007 Sage Publications.

Developing solid particulate vaccine adjuvants - surface bound antigen favouring a humoural response, whereas entrapped antigen shows a tendency for cell mediated immunity

This present study compares the efficacy of microsphere formulations, and their method of antigen presentation, for the delivery of the TB sub-unit vaccine antigen, Ag85B-ESAT-6. Microspheres based on poly(lactide-co-glycolide) (PLGA) and chitosan incorporating dimethyldioctadecylammonium bromide (DDA) were prepared by either the w/o/w double emulsion method (entrapped antigen) or the o/w single emulsion method (surface bound antigen), and characterised for their physico-chemical properties and their ability to promote an immune response to Ag85B-ESAT-6. The method of preparation, and hence method of antigen association, had a pronounced effect on the type of immune response achieved from the microsphere formulations, with surface bound antigen favouring a humoural response, whereas entrapped antigen favoured a cellular response.

Hsc70-induced changes in clathrin-auxilin cage structure suggest a role for clathrin light chains in cage disassembly

The molecular chaperone, Hsc70, together with its cofactor, auxilin, facilitates the ATP-dependent removal of clathrin during clathrin-mediated endocytosis in cells. We have used cryo-electron microscopy to determine the 3D structure of a complex of clathrin, auxilin401-910 and Hsc70 at pH 6 in the presence of ATP, frozen within 20 seconds of adding Hsc70 in order to visualize events that follow the binding of Hsc70 to clathrin and auxilin before clathrin disassembly. In this map,we observe density beneath the vertex of the cage that we attribute to bound Hsc70. This density emerges asymmetrically from the clathrin vertex, suggesting preferential binding by Hsc70 for one of the three possible sites at the vertex. Statistical comparison with a map of whole auxilin and clathrin previously published by us reveals the location of statistically significant differences which implicate involvement of clathrin light chains in structural rearrangements which occur after Hsc70 is recruited. Clathrin disassembly assays using light scattering suggest that loss of clathrin light chains reduces the efficiency with which auxilin facilitates this reaction. These data support a regulatory role for clathrin light chains in clathrin disassembly in addition to their established role in regulating clathrin assembly. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Non-English-speaking defendants in the magistrates court:a comparative study of face-to-face and prison video link interpreter-mediated hearings in England

This study compares interpreter-mediated face-to-face Magistrates Court hearings with those conducted through prison video link in which interpreters are located in court and non- English-speaking defendants in prison. It seeks to examine the impact that the presence of video link has on court actors in terms of interaction and behaviour. The data comprises 11 audio-recordings of face-to-face hearings, 10 recordings of prison video link hearings, semistructured interviews with 27 court actors, and ethnographic observation of hearings as viewed by defendants in Wormwood Scrubs prison in London. The over-arching theme is the pervasive influence of the ecology of the courtroom upon all court actors in interpretermediated hearings and thus on the communication process. Close analysis of the court transcripts shows that their relative proximity to one another can be a determinant of status, interpreting role, mode and volume. The very few legal protocols which apply to interpretermediated cases (acknowledging and ratifying the interpreter, for example), are often forgotten or dispensed with. Court interpreters lack proper training in the specific challenges of court interpreting, whether they are co-present with the defendant or not. Other court actors often misunderstand the interpreter’s role. This has probably come about because courts have adjusted their perceptions of what they think interpreters are supposed to do based on their own experiences of working with them, and have gradually come to accept poor practice (the inability to perform simultaneous interpreting, for example) as the norm. In video link courts, mismatches of sound and image due to court clerks’ failure to adequately track current speakers, poor image and sound quality and the fact that non-English-speaking defendants in pre-and post-court consultations can see and hear interpreters but not their defence advocates are just some of the additional layers of disadvantage and confusion already suffered by non- English-speaking defendants. These factors make it less likely that justice will be done.

Pharmacy (ISSN 2226-4787):a journal of pharmacy education and practice

A journal of pharmacy education and practice is an international scientific open access journal on pharmacy education and practice, and is published by MDPI online quarterly. The practice of pharmacy is changing at an unprecedented rate as the profession moves from a focus upon preparation and supply of medicines to a clinical patient-facing role. While an understanding of the science related to medicines remains core to pharmacy education, the changes in practice are driving changes to the traditional methods of pharmacy education. This is reflected at an international level by major changes in the educational standards set by statutory regulators and by policy statements from bodies such as the World Health Organisation. These changes reflect an increasing trend to look at educational policy at a supra-national level, typified by the “Pharmine Project” led by the Association of European Faculties of Pharmacy.

Exploring the correlation between lipid packaging in lipoplexes and their transfection efficacy

Whilst there is a large body of evidence looking at the design of cationic liposomes as transfection agents, correlates of formulation to function remain elusive. In this research, we investigate if lipid packaging can give further insights into transfection efficacy. DNA lipoplexes composed of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) or 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) in combination with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or 1,2-stearoyl-3-trimethylammonium-propane (DSTAP) were prepared by the lipid hydration method. Each of the formulations was prepared by hydration in dH2O or phosphate buffer saline (PBS) to investigate the effect of buffer salts on lipoplex physicochemical characteristics and in vitro transfection. In addition, Langmuir monolayer studies were performed to investigate any possible correlation between lipid packaging and liposome attributes. Using PBS, rather than dH2O, to prepare the lipoplexes increased the size of vesicles in most of formulations and resulted in variation in transfection efficacies. However, one combination of lipids (DSPE:DOTAP) could not form liposomes in PBS, whilst the DSPE:DSTAP combination could not form liposomes in either aqueous media. Monolayer studies demonstrated saturated lipid combinations offered dramatically closer molecular packing compared to the other combinations which could suggest why this lipid combination could not form vesicles. Of the lipoplexes prepared, those formulated with DSTAP showed higher transfection efficacy, however, the effect of buffer on transfection efficiency was formulation dependent. © 2011 by the authors; licensee MDPI, Basel, Switzerland.

The influence of formulation and manufacturing process parameters on the characteristics of lyophilized orally disintegrating tablets

Gelatin is a principal excipient used as a binder in the formulation of lyophilized orally disintegrating tablets. The current study focuses on exploiting the physicochemical properties of gelatin by varying formulation parameters to determine their influence on orally disintegrating tablet (ODT) characteristics. Process parameters, namely pH and ionic strength of the formulations, and ball milling were investigated to observe their effects on excipient characteristics and tablet formation. The properties and characteristics of the formulations and tablets which were investigated included: glass transition temperature, wettability, porosity, mechanical properties, disintegration time, morphology of the internal structure of the freeze-dried tablets, and drug dissolution. The results from the pH study revealed that adjusting the pH of the formulation away from the isoelectric point of gelatin, resulted in an improvement in tablet disintegration time possibly due to increase in gelatin swelling resulting in greater tablet porosity. The results from the ionic strength study revealed that the inclusion of sodium chloride influenced tablet porosity, tablet morphology and the glass transition temperature of the formulations. Data from the milling study showed that milling the excipients influenced formulation characteristics, namely wettability and powder porosity. The study concludes that alterations of simple parameters such as pH and salt concentration have a significant influence on formulation of ODT. © 2011 by the authors; licensee MDPI, Basel, Switzerland.

Radiolabelling of antigen and liposomes for vaccine biodistribution studies

A relatively simple and effective method to follow the movement of pharmaceutical preparations such as vaccines in biodistribution studies is to radiolabel the components. Whilst single radiolabelling is common practice, in vaccine systems containing adjuvants the ability to follow both the adjuvant and the antigen is favourable. To this end, we have devised a dual-radiolabelling method whereby the adjuvant (liposomes) is labelled with 3H and the antigen (a subunit protein) with 125I. This model is stable and reproducible; we have shown release of the radiolabels to be negligible over periods of up to 1 week in foetal calf serum at 37° C. In this paper we describe the techniques which enable the radiolabelling of various components, assessing stability and processing of samples which all for their application in biodistribution studies. Furthermore we provide examples derived from our studies using this model in tuberculosis vaccine biodistribution studies. © 2010 by the authors.

Investigation of formulation and process of lyophilised orally disintegrating tablet (ODT) using novel amino acid combination

Lyophilised orally disintegrating tablets (ODTs) have achieved a great success in overcoming dysphagia associated with conventional solid dosage forms. However, the extensive use of saccharides within the formulation limits their use in treatment of chronic illnesses. The current study demonstrates the feasibility of using combination of proline and serine to formulate zero sacharide ODTs and investigates the effect of freezing protocol on sublimation rate and tablets characteristics. The results showed that inclusion of proline and serine improved ODT properties when compared to individual counterparts. Additionally, annealing the ODTs facilitated the sublimation process and shortened the disintegration time. © 2010 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland.

Pharmaceutics:a new open-access journal for all those interested in designing medicines

We know the many hurdles that face us when we look to deliver a drug, starting from the basic characteristics of the drug (its solubility, stability, absorption and biodistribution), to overcoming the physiological barriers faced in reaching the target site, and to maintaining the concentration within the therapeutic window. In addition we must also remember the patient needs in this – is it a child that needs a liquid dosage form? Is it someone having to take multiple doses in a day? Do we need a rapid onset of action in a convenient format? Will people find it convenient to take the drug in the format we are presenting to them – or are there alternative options? [...]

Utopien, Epiphanien und Melancholie:Der Norden als Erfahrungs- und Imaginationsraum in der deutschsprachigen Gegenwartsliteratur

Die lange zu bemerkende „markante Absenz der nordischen Hemisphäre als literarischer Raum“ (Uwe Schütte) in der deutschsprachigen Nachkriegsliteratur hat in den letzten Jahren ein Ende gefunden. In Texten so unterschiedlicher Autoren wie Sybille Berg, Klaus Böldl, Melitta Breznik, Judith Hermann, Peter Stamm und Antje Ravic Strubel wird vermehrt der (skandinavische) Norden als literarischer Schauplatz in Form eines schriftstellerischen Imaginationsraums utopischer Qualität gestaltet. Allen Texten ist dabei gemein, dass der Norden als bewusst aufgesuchte Peripherie aufgefasst wird, die im Gegensatz zum mitteleuropäischen Zentrum steht. In der postmodernen Welt der Globalisierung, die sich mit Baudrillard als eine die Wirklichkeit usurpierende Simulation begreifen lässt, bietet der Norden den typischerweise mit melancholischen Zügen gezeichneten, an Entfremdung von ihrer Umwelt leidenden Figuren als Ort räumlicher Extremität die zugleich reale und befremdende Erfahrung einer Natur der Extreme, die trotz der zunehmenden Homogenisierung der vernetzten Welt Fortbestand hat und die den auch im Norden anzutreffenden bedeutungsleeren „Nicht-Orten“ (Marc Augé) der modernen Transitgesellschaft ein Erlebnis von Sinnhaftigkeit und Ursprünglichkeit gegenüber stellt. Obwohl (besonders bei Judith Hermann) Zweifel geübt werden an der Zugänglichkeit und emotionalen Wirksamkeit dieser Naturphänomene, besitzen sie dennoch das Potenzial bei den auf der Suche nach ihrer Identität und nach Selbsterfahrung befindlichen Figuren Erlebnisse von Epiphanien auszulösen. Lässt sich in einigen der Texte (besonders bei Peter Stamm) eine Auflösung der geographischen Umgebung ins Unbestimmte bzw. (bei Judith Hermann) eine bewusste Verfremdung skandinavischer Topographie verzeichnen, so werden bei Antje Ravic Strubel gar die Zuschreibungen von Geschlecht und gender vor nördlicher Kulisse aufgebrochen und es entsteht im Norden sowohl ein Freiraum geschlechtlicher Performanz (Judith Butler) als auch ein allgemeiner Projektionsraum für mögliche Lebensentwürfe – und für eine Aufarbeitung der deutschen Geschichte in Skandinavien, die noch deutlicher bei Melitta Breznik anklingt und den Norden auch als Chronotopos und postkolonialen Raum identifiziert. Ausgehend von diesen grundlegenden Merkmalen der Texte geht dieser Beitrag der Frage nach, wie vor dem Hintergrund des viel beschworenen spatial turn neue Perspektiven auf die aktuelle intensive literarische Auseinandersetzung mit dem Norden als konkreter Ort geographischer Realität und als räumliche Metapher mit epistemologischer Aufladung zu gewinnen sind.

Characterization and optimization of bilosomes for oral vaccine delivery

Oral vaccines offer significant benefits due to the ease of administration, better patient compliance and non-invasive, needle-free administration. However, this route is marred by the harsh gastro intestinal environment which is detrimental to many vaccine formats. To address this, a range of delivery systems have been considered including bilosomes; these are bilayer vesicles constructed from non-ionic surfactants combined with the inclusion of bile salts which can stabilize the vesicles in the gastro intestinal tract by preventing membrane destabilization. The aim of this study was to investigate the effect of formulation parameters on bilosome carriers using Design of Experiments to select an appropriate formulation to assess in vivo. Bilosomes were constructed from monopalmitoylglycerol, cholesterol, dicetyl phosphate and sodium deoxycholate at different blends ratios. The optimized bilosome formulation was identified and the potential of this formulation as an oral vaccine delivery system were assessed in biodistribution and vaccine efficacy studies. Results showed that the larger bilosomes vesicles (~6 µm versus 2 µm in diameter) increased uptake within the Peyer's patches and were able to reduce median temperature differential change and promote a reduction in viral cell load in an influenza challenge study. © 2013 Informa UK, Ltd.

Effectiveness and uptake of screening programmes for coronary heart disease and diabetes:a realist review of design components used in interventions

Objective - To evaluate behavioural components and strategies associated with increased uptake and effectiveness of screening for coronary heart disease and diabetes with an implementation science focus. Design - Realist review. Data sources - PubMed, Web of Knowledge, Cochrane Database of Systematic Reviews, Cochrane Controlled Trials Register and reference chaining. Searches limited to English language studies published since 1990. Eligibility criteria - Eligible studies evaluated interventions designed to increase the uptake of cardiovascular disease (CVD) and diabetes screening and examined behavioural and/or strategic designs. Studies were excluded if they evaluated changes in risk factors or cost-effectiveness only. Results - In 12 eligible studies, several different intervention designs and evidence-based strategies were evaluated. Salient themes were effects of feedback on behaviour change or benefits of health dialogues over simple feedback. Studies provide mixed evidence about the benefits of these intervention constituents, which are suggested to be situation and design specific, broadly supporting their use, but highlighting concerns about the fidelity of intervention delivery, raising implementation science issues. Three studies examined the effects of informed choice or loss versus gain frame invitations, finding no effect on screening uptake but highlighting opportunistic screening as being more successful for recruiting higher CVD and diabetes risk patients than an invitation letter, with no differences in outcomes once recruited. Two studies examined differences between attenders and non-attenders, finding higher risk factors among non-attenders and higher diagnosed CVD and diabetes among those who later dropped out of longitudinal studies. Conclusions - If the risk and prevalence of these diseases are to be reduced, interventions must take into account what we know about effective health behaviour change mechanisms, monitor delivery by trained professionals and examine the possibility of tailoring programmes according to contexts such as risk level to reach those most in need. Further research is needed to determine the best strategies for lifelong approaches to screening.

Th1 immune responses can be modulated by varying dimethyldioctadecylammonium and distearoyl-sn-glycero-3-phosphocholine content in liposomal adjuvants

Objectives - Cationic liposomes of dimethyldioctadecylammonium bromide (DDA) combined with trehalose 6,6'-dibehenate (TDB) elicit strong cell-mediated and antibody immune responses; DDA facilitates antigen adsorption and presentation while TDB potentiates the immune response. To further investigate the role of DDA, DDA was replaced with the neutral lipid of distearoyl-sn-glycero-3-phosphocholine (DSPC) over a series of concentrations and these systems investigated as adjuvants for the delivery of Ag85B–ESAT-6-Rv2660c, a multistage tuberculosis vaccine. Methods - Liposomal were prepared at a 5?:?1 DDA–TDB weight ratio and DDA content incrementally replaced with DSPC. The physicochemical characteristics were assessed (vesicle size, zeta potential and antigen loading), and the ability of these systems to act as adjuvants was considered. Key findings - As DDA was replaced with DSPC within the liposomal formulation, the cationic nature of the vesicles decreases as does electrostatically binding of the anionic H56 antigen (Hybrid56; Ag85B-ESAT6-Rv2660c); however, only when DDA was completed replaced with DSPC did vesicle size increase significantly. T-helper 1 (Th1)-type cell-mediated immune responses reduced. This reduction in responses was attributed to the replacement of DDA with DSPC rather than the reduction in DDA dose concentration within the formulation. Conclusion - These results suggest Th1 responses can be controlled by tailoring the DDA/DSPC ratio within the liposomal adjuvant system.