Liposomal delivery of p-ialB and p-omp25 DNA vaccines improves immunogenicity but fails to provide full protection against B. melitensis challenge

BACKGROUND: We have previously demonstrated protective efficacy against B. melitensis using formulations of naked DNA vaccines encoding genes ialB and omp25. The present study was undertaken to further understand the immune response generated by the protective vaccination regimens and to evaluate cationic liposome adsorption as a delivery method to improve vaccine utility. METHODS: The protective efficacy and immunogenicity of vaccines delivered as four doses of naked DNA, a single dose of naked DNA or a single dose of DNA surface adsorbed to cationic liposomes were compared using the BALB/c murine infection model of B. melitensis. Antigen-specific T cells and antibody responses were compared between the various formulations. RESULTS: The four dose vaccination strategy was confirmed to be protective against B. melitensis challenge. The immune response elicited by the various vaccines was found to be dependent upon both the antigen and the delivery strategy, with the IalB antigen favouring CD4+ T cell priming and Omp25 antigen favouring CD8+. Delivery of the p-ialB construct as a lipoplex improved antibody generation in comparison to the equivalent quantity of naked DNA. Delivery of p-omp25 as a lipoplex altered the profile of responsive T cells from CD8+ to CD4+ dominated. Under these conditions neither candidate delivered by single dose naked DNA or lipoplex vaccination methods was able to produce a robust protective effect. CONCLUSIONS: Delivery of the p-omp25 and p-ialB DNA vaccine candidates as a lipoplex was able to enhance antibody production and effect CD4+ T cell priming, but was insufficient to promote protection from a single dose of either vaccine. The enhancement of immunogenicity by lipoplex delivery is a promising step toward improving the practicality of these two candidate vaccines, and suggests that this lipoplex formulation may be of value in situations where improvements to CD4+ responses are required. However, in the case of Brucella vaccine development it is suggested that further modifications to the candidate vaccines and delivery strategies will be required in order to deliver sustained protection

The analysis of para-cresol production and tolerance in Clostridium difficile 027 and 012 strains

<p>Abstract</p> <p>Background</p> <p><it>Clostridium difficile </it>is the major cause of antibiotic associated diarrhoea and in recent years its increased prevalence has been linked to the emergence of hypervirulent clones such as the PCR-ribotype 027. Characteristically, <it>C. difficile </it>infection (CDI) occurs after treatment with broad-spectrum antibiotics, which disrupt the normal gut microflora and allow <it>C. difficile </it>to flourish. One of the relatively unique features of <it>C. difficile </it>is its ability to ferment tyrosine to <it>para</it>-cresol via the intermediate <it>para</it>-hydroxyphenylacetate (<it>p-</it>HPA). <it>P</it>-cresol is a phenolic compound with bacteriostatic properties which <it>C. difficile </it>can tolerate and may provide the organism with a competitive advantage over other gut microflora, enabling it to proliferate and cause CDI. It has been proposed that the <it>hpdBCA </it>operon, rarely found in other gut microflora, encodes the enzymes responsible for the conversion of <it>p-</it>HPA to <it>p</it>-cresol.</p> <p>Results</p> <p>We show that the PCR-ribotype 027 strain R20291 quantitatively produced more <it>p</it>-cresol <it>in-vitro </it>and was significantly more tolerant to <it>p</it>-cresol than the sequenced strain 630 (PCR-ribotype 012). Tyrosine conversion to <it>p</it>-HPA was only observed under certain conditions. We constructed gene inactivation mutants in the <it>hpdBCA </it>operon in strains R20291 and 630Δ<it>erm </it>which curtails their ability to produce <it>p</it>-cresol, confirming the role of these genes in <it>p-</it>cresol production. The mutants were equally able to tolerate <it>p</it>-cresol compared to the respective parent strains, suggesting that tolerance to <it>p</it>-cresol is not linked to its production.</p> <p>Conclusions</p> <p><it>C. difficile </it>converts tyrosine to <it>p</it>-cresol, utilising the <it>hpdBCA </it>operon in <it>C. difficile </it>strains 630 and R20291. The hypervirulent strain R20291 exhibits increased production of and tolerance to <it>p-</it>cresol, which may be a contributory factor to the virulence of this strain and other hypervirulent PCR-ribotype 027 strains.</p

Suramin: Effectiveness of analogues reveals structural features that are important for the potent trypanocidal activity of the drug

Suramin was the first effective drug for the treatment of human African sleeping sickness. Structural analogues of the trypanocide have previously been shown to be potent inhibitors of several enzymes. Therefore, four suramin analogues lacking the methyl group on the intermediate rings and with different regiochemistry of the naphthalenetrisulphonic acid groups and the phenyl rings were tested to establish whether they exhibited improved antiproliferative activity against bloodstream forms of Trypanosomes brucei compared to the parent compound. The four analogues exhibited low trypanocidal activity and weak inhibition of the antitrypanosomal activity of suramin in competition experiments. This indicates that the strong trypanocidal activity of suramin is most likely due to the presence of methyl groups on its intermediate rings and to the specific regiochemistry of naphthalenetrisulphonic acid groups. These two structural features are also likely to be important for the inhibition mechanism of suramin because DNA distribution and nucleus/kinetoplast configuration analyses suggest that the analogues inhibit mitosis while suramin inhibits cytokinesis

Suramin inhibits osteoarthritic cartilage degradation by increasing extracellular levels of chondroprotective tissue inhibitor of metalloproteinases 3

Osteoarthritis is a common degenerative joint disease for which no disease-modifying drugs are currently available. Attempts to treat the disease with small molecule inhibitors of the metalloproteinases that degrade the cartilage matrix have been hampered by a lack of specificity. We aimed to inhibit cartilage degradation by augmenting levels of the endogenous metalloproteinase inhibitor, tissue inhibitor of metalloproteinases 3 (TIMP-3), through blocking its interaction with the endocytic scavenger receptor, low-density lipoprotein receptor-related protein 1 (LRP1). We discovered that suramin (C51H40N6O23S6) bound to TIMP-3 with a KD value of 1.9 ± 0.2 nM and inhibited its endocytosis via LRP1, thus increasing extracellular levels of TIMP-3 and inhibiting cartilage degradation by the TIMP-3 target enzyme, adamalysin with thrombospondin motifs 5 (ADAMTS-5). NF279, a structural analogue of suramin, has increased affinity for TIMP-3 and increased ability to inhibit TIMP- 3 endocytosis and protect cartilage. Suramin is thus a promising scaffold for the development of novel therapeutics to increase TIMP-3 levels and inhibit cartilage degradation in osteoarthritis

GRB 090417B and its Host Galaxy: A Step Towards an Understanding of Optically-Dark Gamma-Ray Bursts

GRB 090417B was an unusually long burst with a T_90 duration of at least 2130 s and a multi-peaked light curve at energies of 15-150 keV. It was optically dark and has been associated with a bright star-forming galaxy at a redshift of 0.345 that is broadly similar to the Milky Way. This is one of the few cases where a host galaxy has been clearly identified for a dark gamma-ray burst and thus an ideal candidate for studying the origin of dark bursts. We find that the dark nature of GRB 090417B cannot be explained by high redshift, incomplete observations, or unusual physics in the production of the afterglow. Assuming the standard relativistic fireball model for the afterglow we find that the optical flux is at least 2.5 mag fainter than predicted by the X-ray flux. The Swift/XRT X -ray data are consistent with the afterglow being obscured by a dense, localized sheet of dust approximately 30-80 pc from the burst along the line of sight. Our results suggest that this dust sheet imparts an extinction of A_V >~ 12 mag, which is sufficient to explain the missing optical flux. GRB 090417B is an example of a gamma-ray burst that is dark due to the localized dust structure in its host galaxy.Comment: Accepted for publication in Ap

Automated annotation of chemical names in the literature with tunable accuracy

<p>Abstract</p> <p>Background</p> <p>A significant portion of the biomedical and chemical literature refers to small molecules. The accurate identification and annotation of compound name that are relevant to the topic of the given literature can establish links between scientific publications and various chemical and life science databases. Manual annotation is the preferred method for these works because well-trained indexers can understand the paper topics as well as recognize key terms. However, considering the hundreds of thousands of new papers published annually, an automatic annotation system with high precision and relevance can be a useful complement to manual annotation.</p> <p>Results</p> <p>An automated chemical name annotation system, MeSH Automated Annotations (MAA), was developed to annotate small molecule names in scientific abstracts with tunable accuracy. This system aims to reproduce the MeSH term annotations on biomedical and chemical literature that would be created by indexers. When comparing automated free text matching to those indexed manually of 26 thousand MEDLINE abstracts, more than 40% of the annotations were false-positive (FP) cases. To reduce the FP rate, MAA incorporated several filters to remove "incorrect" annotations caused by nonspecific, partial, and low relevance chemical names. In part, relevance was measured by the position of the chemical name in the text. Tunable accuracy was obtained by adding or restricting the sections of the text scanned for chemical names. The best precision obtained was 96% with a 28% recall rate. The best performance of MAA, as measured with the F statistic was 66%, which favorably compares to other chemical name annotation systems.</p> <p>Conclusions</p> <p>Accurate chemical name annotation can help researchers not only identify important chemical names in abstracts, but also match unindexed and unstructured abstracts to chemical records. The current work is tested against MEDLINE, but the algorithm is not specific to this corpus and it is possible that the algorithm can be applied to papers from chemical physics, material, polymer and environmental science, as well as patents, biological assay descriptions and other textual data.</p

Real time Raman imaging to understand dissolution performance of amorphous solid dispersions

We have employed for the first time Raman spectroscopic imaging along with multi-variate curve resolution (MCR) analysis to investigate in real time and in-situ the dissolution mechanisms that underpin amorphous solid dispersions, with data being collected directly from the dosage form itself. We have also employed a novel rotating disk dissolution rate (RDDR) methodology to track, through the use of high-performance liquid chromatography (HPLC), the dissolution trends of both drug and polymer simultaneously in multi-component systems. Two formulations of poorly water-soluble felodipine in a polymeric matrix of copovidone VA64 which have different drug loadings of 5% and 50% w/w were used as models with the aim of studying the effects of increasing the amount of active ingredient on the dissolution performance. It was found that felodipine and copovidone in the 5% dispersion dissolve with the same dissolution rate and that no Raman spectral changes accompanied the dissolution, indicating that the two components dissolve as single entity, whose behaviour is dominated by water-soluble copovidone. For the 50% drug-loaded dispersion, partial RDDR values of both felodipine and copovidone were found to be extremely low. MCR Raman maps along with classical Raman/X-ray powder diffraction (XRPD) characterisation revealed that after an initial loss of copovidone from the extrudate the drug re-crystallises, pointing to a release dynamics dependent on the low water solubility and high hydrophobicity of felodipine. Raman imaging revealed different rates of transition from amorphous to crystalline felodipine at different locations within the dosage form

Preparation of novel optical fibre-based Cocaine sensors using a molecular imprinted polymer approach

Novel chemical sensors using fibre optic-based techniques for the detection of Cocaine have been developed, utilising molecularly imprinted polymers (MIPs) containing fluorescein moieties as the signalling groups. The fluorescent MIPs were formed and covalently attached to the distal end of specially chosen optical fibres to create fibre optic probe-based sensors. These sensors exhibited are producible and quantifiable change in the intensity of the fluorescence signal received from the sensor in response to Cocaine in aqueous acetonitrile mixtures. High selectivity for Cocaine over Codeine and a range of known Cocaine interferants has been demonstrated for one of the sensors developed in this work