Formulation of polylactide-co-glycolic acid nanospheres for encapsulation and sustained release of poly(ethylene imine)-poly(ethylene glycol) copolymers complexed to oligonucleotides

Antisense oligonucleotides (AOs) have been shown to induce dystrophin expression in muscles cells of patients with Duchenne Muscular Dystrophy (DMD) and in the mdx mouse, the murine model of DMD. However, ineffective delivery of AOs limits their therapeutic potential. Copolymers of cationic poly(ethylene imine) (PEI) and non-ionic poly(ethylene glycol) (PEG) form stable nanoparticles when complexed with AOs, but the positive surface charge on the resultant PEG-PEI-AO nanoparticles limits their biodistribution. We adapted a modified double emulsion procedure for encapsulating PEG-PEI-AO polyplexes into degradable polylactide-co-glycolic acid (PLGA) nanospheres. Formulation parameters were varied including PLGA molecular weight, ester end-capping, and sonication energy/volume. Our results showed successful encapsulation of PEG-PEI-AO within PLGA nanospheres with average diameters ranging from 215 to 240 nm. Encapsulation efficiency ranged from 60 to 100%, and zeta potential measurements confirmed shielding of the PEG-PEI-AO cationic charge. Kinetic measurements of 17 kDa PLGA showed a rapid burst release of about 20% of the PEG-PEI-AO, followed by sustained release of up to 65% over three weeks. To evaluate functionality, PEG-PEI-AO polyplexes were loaded into PLGA nanospheres using an AO that is known to induce dystrophin expression in dystrophic mdx mice. Intramuscular injections of this compound into mdx mice resulted in over 300 dystrophin-positive muscle fibers distributed throughout the muscle cross-sections, approximately 3.4 times greater than for injections of AO alone. We conclude that PLGA nanospheres are effective compounds for the sustained release of PEG-PEI-AO polyplexes in skeletal muscle and concomitant expression of dystrophin, and may have translational potential in treating DMD

The oral microbiome of denture wearers is influenced by natural dentition

Objectives: The composition of dental plaque has been well defined, whereas currently there is limited understanding of the composition of denture plaque and how it directly influences denture related stomatitis (DS). The aims of this study were to compare the microbiomes of denture wearers, and to understand the implications of these towards inter-kingdom and host-pathogen interactions within the oral cavity. Methods: Swab samples were obtained from 123 participants wearing either a complete or partial denture; the bacterial composition of each sample was determined using bar-coded illumina MiSeq sequencing of the bacterial hypervariable V4 region of 16S rDNA. Sequencing data processing was undertaken using QIIME, clustered in Operational Taxonomic Units (OTUs) and assigned to taxonomy. The dentures were sonicated to remove the microbial flora residing on the prosthesis, sonicate was then cultured using diagnostic colorex Candida media. Samples of unstimulated saliva were obtained and antimicrobial peptides (AMP) levels were measured by ELISA. Results: We have shown that dental and denture plaques are significantly distinct both in composition and diversity and that the oral microbiome composition of a denture wearer is variable and is influenced by the location within the mouth. Dentures and mucosa were predominantly made up of Bacilli and Actinobacteria. Moreover, the presence of natural teeth has a significant impact on the overall microbial composition, when compared to the fully edentulous. Furthermore, increasing levels of Candida spp. positively correlate with Lactobacillus spp. AMPs were quantified, though showed no specific correlations. Conclusions: This is the first study to provide a detailed understanding of the oral microbiome of denture wearers and has provided evidence that DS development is more complex than simply a candidal infection. Both fungal and bacterial kingdoms clearly play a role in defining the progression of DS, though we were unable to show a defined role for AMPs

Communicating the uncertainty in estimated greenhouse gas emissions from agriculture

In an effort to mitigate anthropogenic effects on the global climate system, industrialised countries are required to quantify and report, for various economic sectors, the annual emissions of greenhouse gases from their several sources and the absorption of the same in different sinks. These estimates are uncertain, and this uncertainty must be communicated effectively, if government bodies, research scientists or members of the public are to draw sound conclusions. Our interest is in communicating the uncertainty in estimates of greenhouse gas emissions from agriculture to those who might directly use the results from the inventory. We tested six methods of communication. These were: a verbal scale using the IPCC calibrated phrases such as ‘likely’ and ‘very unlikely’; probabilities that emissions are within a defined range of values; confidence intervals for the expected value; histograms; box plots; and shaded arrays that depict the probability density of the uncertain quantity. In a formal trial we used these methods to communicate uncertainty about four specific inferences about greenhouse gas emissions in the UK. Sixty four individuals who use results from the greenhouse gas inventory professionally participated in the trial, and we tested how effectively the uncertainty about these inferences was communicated by means of a questionnaire. Our results showed differences in the efficacy of the methods of communication, and interactions with the nature of the target audience. We found that, although the verbal scale was thought to be a good method of communication it did not convey enough information and was open to misinterpretation. Shaded arrays were similarly criticised for being open to misinterpretation, but proved to give the best impression of uncertainty when participants were asked to interpret results from the greenhouse gas inventory. Box plots were most favoured by our participants largely because they were particularly favoured by those who worked in research or had a stronger mathematical background. We propose a combination of methods should be used to convey uncertainty in emissions and that this combination should be tailored to the professional grou

Increased usage of special educational services by children born to mothers with systemic lupus erythematosus and antiphospholipid antibodies

Introduction: Surveys of long-term health and developmental outcomes of children born to mothers with systemic lupus erythematosus (SLE) have suggested an increase in learning disabilities among these children. We performed this observational study to investigate the relationship between maternal autoantibodies and antiphospholipid antibody syndrome (APS) in maternal lupus patients and neurocognitive development among their offspring. Methods: SLE mothers with at least one live birth postlupus diagnosis were enrolled. Data on maternal medical/obstetric history and children’s perinatal/ medical history were collected by structured interview and medical record reviews. The primary outcome was requirement for special educational (SE) services, a proxy for developmental delays. Multiple logistic regression modelling was used to examine associations between APS and autoantibodies with SE usage, accounting for SLE disease severity and potential confounders. Results: Data on 38 mothers and 60 offspring were analysed: SE service usage was reported for 15 of 60 (25%) offspring. Maternal APS history was significantly associated with increased use of SE services among offspring, including after adjustment for lupus anticoagulant (LA) positivity and potential confounders (OR 5.5–9.4 for delays age ≥2; p<0.05). The presence of LA, but not other antiphospholipid antibodies, was also associated with increased SE services usage. Conclusions: Maternal APS and LA were independently associated with increased usage of special educational services among offspring of women with SLE.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/108201/1/Lupus Sci Med-2014-Marder-.pdf5

Managing ‘academic value’: the 360-degree perspective

The raison d'etre of all universities is to create and deliver ‘academic value’, which we define as the sum total of the contributions from the 360-degree ‘angles’ of the academic community, including all categories of staff, as well as external stakeholders (e.g. regulatory, commercial, professional and community interests). As a way to conceptualise these complex relationships, we present the ‘academic wheel’ to illustrate the structural nature of them. We then discuss the implications of the different – and sometimes difficult – perspectives of academic, professional and administrative groups in the context of a number of important social psychological processes. We ask whether it is possible to reconcile, what is sometimes perceived as, managerial Taylorism with the academic freedom of (Laurie) Taylorism. We conclude that recognition and active management of these processes are required for each university to optimise its own brand of core academic value

Representability problems for coarse-grained water potentials

The use of an effective intermolecular potential often involves a compromise between more accurate, complex functional forms and more tractable simple representations. To study this choice in detail, we systematically derive coarse-grained isotropic pair potentials that accurately reproduce the oxygen-oxygen radial distribution function of the TIP4P-Ew water model at state points over density ranges from 0.88-1.30g/cc and temperature ranges from 235K-310K. Although by construction these effective potentials correctly represent the isothermal compressibility of TIP4P-Ew water, they do not accurately resolve other thermodynamic properties such as the virial pressure, the internal energy or thermodynamic anomalies. Because at a given state point the pair potential that reproduces the pair structure is unique, we have therefore explicitly demonstrated that it is impossible to simultaneously represent the pair-structure and several key equilibrium thermodynamic properties of water with state-point dependent radially symmetric pair potentials. We argue that such representability problems are related to, but different from, more widely acknowledged transferability problems, and discuss in detail the implications this has for the modeling of water and other liquids by coarse-grained potentials. Nevertheless, regardless of thermodynamic inconsistencies, the state-point dependent effective potentials for water do generate structural and dynamical anomalies.Comment: 22 page

Terrestrial environmental change across the onset of the PETM and the associated impact on biomarker proxies:A cautionary tale

The following supplementary information includes one dataset which contains 3 tables: Biomarker distributions and proxies at Cobham, UK Bulk and compound specific isotope data at Cobham (UK) Model-derived mean annual surface temperature and precipitation estimates as a function of CO2 at Cobham (UK)

Nutritional B vitamin deficiency alters the expression of key proteins associated with vascular smooth muscle cell proliferation and migration in the aorta of atherosclerotic apolipoprotein E null mice.

Low B vitamin status is linked with human vascular disease. We employed a proteomic and biochemical approach to determine whether nutritional folate deficiency and/or hyperhomocysteinemia altered metabolic processes linked with atherosclerosis in ApoE null mice. Animals were fed either a control fat (C; 4 % w/w lard) or a high-fat [HF; 21 % w/w lard and cholesterol (0/15 % w/w)] diet with different B vitamin compositions for 16 weeks. Aorta tissue was prepared and global protein expression, B vitamin, homocysteine and lipoprotein status measured. Changes in the expression of aorta proteins were detected in response to multiple B vitamin deficiency combined with a high-fat diet (P < 0.05) and were strongly linked with lipoprotein concentrations measured directly in the aorta adventitia (P < 0.001). Pathway analysis revealed treatment effects in the aorta-related primarily to cytoskeletal organisation, smooth muscle cell adhesion and invasiveness (e.g., fibrinogen, moesin, transgelin, vimentin). Combined B vitamin deficiency induced striking quantitative changes in the expression of aorta proteins in atherosclerotic ApoE null mice. Deregulated expression of these proteins is associated with human atherosclerosis. Cellular pathways altered by B vitamin status included cytoskeletal organisation, cell differentiation and migration, oxidative stress and chronic inflammation. These findings provide new insight into the molecular mechanisms through which B vitamin deficiency may accelerate atherosclerosis