Deletion at ITPR1 Underlies Ataxia in Mice and Spinocerebellar Ataxia 15 in Humans

We observed a severe autosomal recessive movement disorder in mice used within our laboratory. We pursued a series of experiments to define the genetic lesion underlying this disorder and to identify a cognate disease in humans with mutation at the same locus. Through linkage and sequence analysis we show here that this disorder is caused by a homozygous in-frame 18-bp deletion in Itpr1 (Itpr1Δ18/Δ18), encoding inositol 1,4,5-triphosphate receptor 1. A previously reported spontaneous Itpr1 mutation in mice causes a phenotype identical to that observed here. In both models in-frame deletion within Itpr1 leads to a decrease in the normally high level of Itpr1 expression in cerebellar Purkinje cells. Spinocerebellar ataxia 15 (SCA15), a human autosomal dominant disorder, maps to the genomic region containing ITPR1; however, to date no causal mutations had been identified. Because ataxia is a prominent feature in Itpr1 mutant mice, we performed a series of experiments to test the hypothesis that mutation at ITPR1 may be the cause of SCA15. We show here that heterozygous deletion of the 5′ part of the ITPR1 gene, encompassing exons 1–10, 1–40, and 1–44 in three studied families, underlies SCA15 in humans

Hypoxia and Prostaglandin E Receptor 4 Signalling Pathways Synergise to Promote Endometrial Adenocarcinoma Cell Proliferation and Tumour Growth

The prostaglandin endoperoxide synthase (PTGS) pathway is a potent driver of tumour development in humans by enhancing the biosynthesis and signalling of prostaglandin (PG) E2. PTGS2 expression and PGE2 biosynthesis is elevated in endometrial adenocarcinoma, however the mechanism whereby PTGS and PGE2 regulate endometrial tumour growth is unknown. Here we investigated (a) the expression profile of the PGE synthase enzymes (PTGES, PTGES-2, PTGES-3) and PGE receptors (PTGER1–4) in endometrial adenocarcinomas compared with normal endometrium and (b) the role of PTGER4 in endometrial tumorigenesis in vivo. We found elevated expression of PTGES2 and PTGER4 and suppression of PTGER1 and PTGER3 in endometrial adenocarcinomas compared with normal endometrium. Using WT Ishikawa endometrial adenocarcinoma cells and Ishikawa cells stably transfected with the full length PTGER4 cDNA (PTGER4 cells) xenografted in the dorsal flanks of nude mice, we show that PTGER4 rapidly and significantly enhances tumour growth rate. Coincident with enhanced PTGER4-mediated tumour growth we found elevated expression of PTGS2 in PTGER4 xenografts compared with WT xenografts. Furthermore we found that the augmented growth rate of the PTGER4 xenografts was not due to enhanced angiogenesis, but regulated by an increased proliferation index and hypoxia. In vitro, we found that PGE2 and hypoxia independently induce expression of PTGER4 indicating two independent pathways regulating prostanoid receptor expression. Finally we have shown that PGE2 and hypoxia synergise to promote cellular proliferation of endometrial adenocarcinoma cells

Assessment of alcohol problems using AUDIT in a prison setting: more than an 'aye or no' question

<br>Background: Alcohol problems are a major UK and international public health issue. The prevalence of alcohol problems is markedly higher among prisoners than the general population. However, studies suggest alcohol problems among prisoners are under-detected, under-recorded and under-treated. Identifying offenders with alcohol problems is fundamental to providing high quality healthcare. This paper reports use of the AUDIT screening tool to assess alcohol problems among prisoners.</br> <br>Methods: Universal screening was undertaken over ten weeks with all entrants to one male Scottish prison using the AUDIT standardised screening tool and supplementary contextual questions. The questionnaire was administered by trained prison officers during routine admission procedures. Overall 259 anonymised completed questionnaires were analysed.</br> <br>Results: AUDIT scores showed a high prevalence of alcohol problems with 73% of prisoner scores indicating an alcohol use disorder (8+), including 36% having scores indicating ‘possible dependence’ (20-40). AUDIT scores indicating ‘possible dependence’ were most apparent among 18-24 and 40-64 year-olds (40% and 56% respectively). However, individual questions showed important differences, with younger drinkers less likely to demonstrate habitual and addictive behaviours than the older age group. Disparity between high levels of harmful/hazardous/dependent drinking and low levels of ‘treatment’ emerged (only 27% of prisoners with scores indicating ‘possible dependence’ reported being ‘in treatment’). Self-reported associations between drinking alcohol and the index crime were identified among two-fifths of respondents, rising to half of those reporting violent crimes.</br> <br>Conclusions: To our knowledge, this is the first study to identify differing behaviours and needs among prisoners with high AUDIT score ranges, through additional analysis of individual questions. The study has identified high prevalence of alcohol use, varied problem behaviours, and links across drinking, crime and recidivism, supporting the argument for more extensive provision of alcohol-focused interventions in prisons. These should be carefully targeted based on initial screening and assessment, responsive, and include care pathways linking prisoners to community services. Finally, findings confirm the value and feasibility of routine use of the AUDIT screening tool in prison settings, to considerably enhance practice in the detection and understanding of alcohol problems, improving on current more limited questioning (e.g. ‘yes or no’ questions).</br&gt

Motor Subtype as a Predictor of Future Working Memory Performance in Idiopathic Parkinson's Disease

Parkinson’s disease is a progressive neurodegenerative disorder associated with reduced spatial and verbal working memory ability. There are two established motor subtypes of PD, tremor dominant (TD) and postural instability and gait difficulty (PIGD). This study used structural equation modelling to explore the longitudinal relationship between the two subtypes and working memory assessed at a 2-year follow-up. The study comprised 84 males and 30 females (N = 114), aged between 39 and 85 (M = 64.82, SD = 9.23) with confirmed PD. There was no significant relationship between motor subtype at Time 1 and working memory at Time 2. Postural symptom severity at Time 1 predicted Time 2 spatial working memory for the PIGD subtype (p = .011) but not the TD subtype. Tremor symptoms were not associated with Time 2 working memory in either subtype. Predictive significance of Time 1 postural symptoms only in the PIGD subtype suggests an interaction between symptom dominance (subtype) and symptom severity that future subtyping should consider. This study demonstrates a predictive relationship between postural difficulties and working memory performance assessed at a 2-year follow-up. Establishing physical symptoms as predictors of cognitive change could have significant clinical importance

Human cathelicidin production by the cervix

hCAP18/LL-37 is the sole human cathelicidin; a family of host defence peptides with key roles in innate host defence. hCAP18/LL-37 is expressed primarily by neutrophils and epithelial cells, but its production and function in the lower genital tract is largely uncharacterised. Despite the significant roles for cathelicidin in multiple organs and inflammatory processes, its impact on infections that could compromise fertility and pregnancy is unknown. The aim of this study was to investigate cathelicidin production, regulation and function in the cervix. hCAP18/LL-37 was found to be present in cervicovaginal secretions collected from women in the first trimester of pregnancy and to be expressed at significantly higher levels in samples from women with alterations in vaginal bacterial flora characteristic of bacterial vaginosis. In endocervical epithelial cell lines, expression of the gene encoding hCAP18/LL-37 (CAMP) was not affected by TLR agonists, but was found to be up-regulated by both 1, 25 hydroxyvitamin D3 and 25 hydroxyvitamin D3. However, no association was found between serum levels of vitamin D and hCAP18/LL-37 concentrations in cervicovaginal secretions (n = 116). Exposure to synthetic LL-37 had a pro-inflammatory effect on endocervical epithelial cell lines, increasing secretion of inflammatory cytokine IL-8. Together these data demonstrate inducible expression of hCAP18/LL-37 in the female lower reproductive tract in vivo and suggest the capacity for this peptide to modulate host defence to infection in this system. Further investigation will elucidate the effects of hCAP18/LL-37 on the physiology and pathophysiology of labour, and may lead to strategies for the prevention of infection-associated preterm birth

Seawater carbonate chemistry and growth and grazing impact of Antarctic heterotrophic nanoflagellates

High-latitude oceans have been identified as particularly vulnerable to ocean acidification if anthropogenic CO2 emissions continue. Marine microbes are an essential part of the marine food web and are a critical link in biogeochemical processes in the ocean, such as the cycling of nutrients and carbon. Despite this, the response of Antarctic marine microbial communities to ocean acidification is poorly understood. We investigated the effect of increasing fCO2 on the growth of heterotrophic nanoflagellates (HNFs), nano- and picophytoplankton, and prokaryotes (heterotrophic Bacteria and Archaea) in a natural coastal Antarctic marine microbial community from Prydz Bay, East Antarctica. At CO2 levels ≥634 µatm, HNF abundance was reduced, coinciding with increased abundance of picophytoplankton and prokaryotes. This increase in picophytoplankton and prokaryote abundance was likely due to a reduction in top-down control of grazing HNFs. Nanophytoplankton abundance was elevated in the 634 µatm treatment, suggesting that moderate increases in CO2 may stimulate growth. The taxonomic and morphological differences in CO2 tolerance we observed are likely to favour dominance of microbial communities by prokaryotes, nanophytoplankton, and picophytoplankton. Such changes in predator–prey interactions with ocean acidification could have a significant effect on the food web and biogeochemistry in the Southern Ocean, intensifying organic-matter recycling in surface waters; reducing vertical carbon flux; and reducing the quality, quantity, and availability of food for higher trophic levels

Relationship between 25(OH) vitamin D and hCAP18/LL37 in endocervical (END E6/E7) and ectocervical (ECT E6/E7) cell lines.

<p>Correlation between serum 25(OH) vitamin D and CVS hCAP18/LL-37 (r = −0.13; n = 122; p = 0.14). Data presented as scatter plots and divided in to deficient <25 nmol/l, insufficient 25–75 nmol/l and adequate >75 nmol/l. Data analysed by Pearson's Correlation.</p

Effect of LL-37 receptor antagonists on IL-8 secretion in endocervical (END E6/E7) and ectocervical (ECT E6/E7) cell lines.

<p>END E6/E7 cells were pre-treated for 30 minutes with or without antagonists before being cultured for 24 hours with scrambled LL-37 (25 µg/ml) and LL-37 (25 µg/ml). (A) END E6/E7 treated with PTX (GPCR antagonist, 200 ng/ml), (B) ECT E6/E7 cells treated with PTC, (C) END E6/E7 cells treated with WRW4 (FPR2 antagonist, 10 µM), (D) ECT E6/E7 cells treated with WRW4, (E) END E6/E7 cells treated with KN-62 (P2X₇R antagonist, 10 µM), (F) ECT E6/E7 cells treated with KN-62, (G) END E6/E7 cells treated with oATP (P2X₇R antagonist, 100 µM) and (H) ECT E6/E7 cells treated with oATP. Data presented as mean concentration ± SEM (error bars). *, **, ***, p<0.05, 0.01, 0.001 respectively. (2way ANOVA with multiple comparison tests).</p

Cervical hCAP18/LL-37 expression.

<p>(<b>A</b>) Correlation between cervicovaginal hCAP18/LL-37 and Myeloperoxidase (r = 0.1; n = 77; p = 0.3). (<b>B</b>) Representative western blot of hCAP18/LL-37 expression in cervicovaginal secretions. The 18 kDa hCAP18 and the 5–10 kDa cleaved peptide LL-37 were detected in cervicovaginal secretions.</p