Cloning, functional characterisation and population analysis of a variant form of the human glycine type 2 transporter

AbstractTwo forms of glycine transporter have been described to date, GlyT-1 and GlyT-2. The GlyT-2 form is expressed mainly in the spinal cord, brainstem and cerebellum. Here we describe the identification of a variant form of the human GlyT-2 (SC6), showing three amino acid changes to the previously reported protein. Population analysis identified the allele causing one of the polymorphisms, D463N, at 10% within the population with 3% being homozygous for the change. We also transfected our new variant into mammalian cells and compared it to the published cDNA, showing that the three amino acid changes present have no major effect on the biochemical properties of the transporter

Magnetic resonance imaging characteristics in patients with psoriatic arthritis and axial manifestations from the MAXIMISE cohort

Objective The current analysis of the MAXIMISE trial was conducted to investigate the presence of post-inflammatory and degenerative spinal changes and inflammatory changes in spinal processes identified in baseline MRIs and their potential for predicting differential treatment effects in a cohort of PsA patients with axial manifestations. Methods Baseline spinal MRIs from the MAXIMISE trial were re-read to identify additional inflammatory (spinal process), post-inflammatory, and degenerative changes, and investigate the differential treatment effect of these imaging features using logistic regression modelling. Results In addition to bone marrow oedema assessed at primary analysis, spinal process inflammation and post-inflammatory changes evaluated by FAt Spondyloarthritis Spine Score were documented in 11.1% and 20.2% patients, respectively. At least one type of degenerative change was noted in 64% patients, with Pfirrmann grade ≥3 (51.1%) being the most common. Combining primary and re-read MRI findings, 67.1% of patients presented with inflammatory or post-inflammatory changes while 21.2% had degenerative changes alone. Although not statistically significant, post-inflammatory changes were associated with a trend for better efficacy outcomes in terms of ASAS20, ASAS40 and BASDAI50 responses; a trend for worse outcomes was observed in the presence of degenerative changes. Conclusion The current analysis revealed the occurrence of additional inflammatory and post-inflammatory changes suggestive of axial PsA (axPsA) and a trend for better clinical outcomes for patients treated with secukinumab. These results elucidate the imaging characteristics and improve our current understanding of axPsA thereby supporting the interpretation of future trials.Funding. This study was supported by Novartis Pharma AG, Switzerland. Disclosure statement: X.B.: consultancy honoraria and research grants: AbbVie, BMS, Galapagos, Chugai, Eli Lilly, Janssen, MSD, Novartis, Pfizer, Roche, Sandoz and UCB; E.P.: employee of Novartis with Novartis stock; L.C.: grants/research support from AbbVie, Amgen, Celgene, Eli Lilly, Pfizer and Novartis; consultant for AbbVie, Amgen, Boehringer Ingelheim, Bristol-Myers Squibb, Celgene, Eli Lilly, Gilead, Janssen, Novartis, Pfizer and UCB; and speaker for AbbVie, Amgen, Biogen, Celgene, Gilead, Eli Lilly, Janssen, Medac, Novartis, Pfizer and UCB; R.B.: research grants: AbbVie, MSD and Roche; consulting fees: AbbVie, Pfizer, Roche, Bristol-Myers, Janssen, UCB pharma and MSD; speakers bureau: AbbVie, Pfizer, Roche, Bristol-Myers, Janssen, UCB pharma, MSD and Lilly; V.N.C.: research grants/honoraria from AbbVie, Galapagos, Janssen, Lilly, Moonlake, Novartis, Pfizer and UCB; E.O.: employee of Novartis; B.S.: was an employee of Novartis until manuscript submission and currently working as an employee of GKM Gesellschaft fuer Therapieforschung mbH; R.L.: research grants: AbbVie, Novartis, Pfizer and UCB; speakers bureau: AbbVie, AstraZeneca, Bristol Myers Squibb, Celgene, Eli-Lilly, Janssen, Gilead, Galapagos, Glaxo-Smith-Kline, Novartis, Pfizer and UCB. Patient consent for publication: Not required. Acknowledgements. The authors thank the patients and the study investigators who participated in this study. The authors also thank Andrew Franklin (Novartis Pharma AG, Basel, Switzerland) for the valuable review. Medical writing support, under the guidance of the authors, was provided by Dhanya Mukundan and Rajeeb Ghosh, Novartis Healthcare Private Limited, Hyderabad, India

Predictors of response to secukinumab in patients with psoriatic arthritis and axial manifestations: a post-hoc analysis of the MAXIMISE trial

Objectives To investigate patient characteristics predictive of response to secukinumab in patients with psoriatic arthritis (PsA) with axial manifestations.Methods In a post-hoc analysis from the MAXIMISE trial (NCT02721966) in patients with PsA and axial manifestations, we tested the hypothesis that the OR of the effect of treatment on the primary endpoint of the trial (Assessment of SpondyloArthritis international Society (ASAS) 20 responder status at week 12) would be different depending on 12 prespecified predictor variables. We applied a two-model logistic regression approach, a main effects and an interaction model.Results The OR (95% CI) for ASAS20 response for the presence of nail dystrophy was 3.2 (95% CI 0.93 to 10.99) in the secukinumab 150 mg group and 5.0 (95% CI 1.47 to 17.19) in the secukinumab 300 mg group compared with the placebo group (p=0.029). Odds of being a responder were similar in men and women in the secukinumab groups, though men fared worse than women in the placebo group (p=0.039). Current smokers were less likely to be ASAS20 responders compared with never smokers regardless of the treatment group (p=0.589).Conclusion Nail dystrophy was identified as a predictor of response to secukinumab in patients with PsA with axial manifestations in the MAXIMISE trial. These findings may be explained by the nail-entheseal concept as part of the axial phenotype in PsA

Origin of norovirus positive samples analysed in this study.

<p>Origin of norovirus positive samples analysed in this study.</p

Alignment of the deduced amino acid sequencing obtained from sporadic strains detected in Malawi and the UK with prototype strains representative of the epidemic strain variants that emerged between 1987 and 2012.

<p>Successive variants are colour coded and prototype stains are listed chronologically with earliest variants at the top. Study sequences are identified according to origin as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146972#pone.0146972.t001" target="_blank">Table 1</a>. Single amino acid identities of the study strains against prototype strains are colour coded using the colour assigned to each epidemic strain variant.</p

Numbers of GII-4 strains analysed and variant assignation according to phylogentic analysis.

<p>Strains that do not align chronologically with the GII-4 variant circulating during the study period study period are highlighted (bold, italic & underlined) in the table.</p

Population Estimation and Trappability of the European Badger (Meles meles): Implications for Tuberculosis Management.

peer-reviewedEstimates of population size and trappability inform vaccine efficacy modelling and are required for adaptive management during prolonged wildlife vaccination campaigns. We present an analysis of mark-recapture data from a badger vaccine (Bacille Calmette–Gue´ rin) study in Ireland. This study is the largest scale (755 km2) mark-recapture study ever undertaken with this species. The study area was divided into three approximately equal–sized zones, each with similar survey and capture effort. A mean badger population size of 671 (SD: 76) was estimated using a closed-subpopulation model (CSpM) based on data from capturing sessions of the entire area and was consistent with a separate multiplicative model. Minimum number alive estimates calculated from the same data were on average 49–51% smaller than the CSpM estimates, but these are considered severely negatively biased when trappability is low. Population densities derived from the CSpM estimates were 0.82–1.06 badgers km22, and broadly consistent with previous reports for an adjacent area. Mean trappability was estimated to be 34–35% per session across the population. By the fifth capture session, 79% of the adult badgers caught had been marked previously. Multivariable modelling suggested significant differences in badger trappability depending on zone, season and age-class. There were more putatively trap-wary badgers identified in the population than trap-happy badgers, but wariness was not related to individual’s sex, zone or season of capture. Live-trapping efficacy can vary significantly amongst sites, seasons, age, or personality, hence monitoring of trappability is recommended as part of an adaptive management regime during large–scale wildlife vaccination programs to counter biases and to improve efficiencies.Department of Agriculture, Food and the MarineTeagasc Walsh Fellowship Programm

Multiplexed SNP genotyping using the Qbead™ system: a quantum dot-encoded microsphere-based assay

We have developed a new method using the Qbead™ system for high-throughput genotyping of single nucleotide polymorphisms (SNPs). The Qbead system employs fluorescent Qdot™ semiconductor nanocrystals, also known as quantum dots, to encode microspheres that subsequently can be used as a platform for multiplexed assays. By combining mixtures of quantum dots with distinct emission wavelengths and intensities, unique spectral ‘barcodes’ are created that enable the high levels of multiplexing required for complex genetic analyses. Here, we applied the Qbead system to SNP genotyping by encoding microspheres conjugated to allele-specific oligonucleotides. After hybridization of oligonucleotides to amplicons produced by multiplexed PCR of genomic DNA, individual microspheres are analyzed by flow cytometry and each SNP is distinguished by its unique spectral barcode. Using 10 model SNPs, we validated the Qbead system as an accurate and reliable technique for multiplexed SNP genotyping. By modifying the types of probes conjugated to microspheres, the Qbead system can easily be adapted to other assay chemistries for SNP genotyping as well as to other applications such as analysis of gene expression and protein–protein interactions. With its capability for high-throughput automation, the Qbead system has the potential to be a robust and cost-effective platform for a number of applications