Detection of prostate cancer-specific transcripts in extracellular vesicles isolated from post-DRE urine

Background: The measurement of gene expression in post-digital rectal examination (DRE) urine specimens provides a non-invasive method to determine a patient’s risk of prostate cancer. Many currently available assays use whole urine or cell pellets for the analysis of prostate cancer-associated genes, although the use of extracellular vesicles (EVs) has also recently been of interest. We investigated the expression of prostate-, kidney-, and bladder- specific transcripts and known prostate cancer biomarkers in urine EVs. Methods: Cell pellets and EVs were recovered from post-DRE urine specimens, with the total RNA yield and quality determined by Bioanalyzer. The levels of prostate, kidney, and bladder- associated transcripts in EVs were assessed by TaqMan qPCR and targeted sequencing. Results: RNA was more consistently recovered from the urine EV specimens, with over 80% of the patients demonstrating higher RNA yields in the EV fraction as compared to urine cell pellets. The median EV RNA yield of 36.4 ng was significantly higher than the median urine cell pellet RNA yield of 4.8 ng. Analysis of the post-DRE urine EVs indicated that prostate-specific transcripts were more abundant than kidney- or bladder-specific transcripts. Additionally, patients with prostate cancer had significantly higher levels of the prostate cancer-associated genes PCA3 and ERG. Conclusions: Post-DRE urine EVs are a viable source of prostate-derived RNAs for biomarker discovery and prostate cancer status can be distinguished from analysis of these specimens. Continued analysis of urine EVs offers the potential discovery of novel biomarkers for pre- biopsy prostate cancer detection

Structural and functional determination of homologs of the Mycobacterium tuberculosis N-acetylglucosamine-6-phosphate deacetylase (NagA)

The (Mtb) pathogen encodes an -acetylglucosamine-6-phosphate deacetylase enzyme, NagA (Rv3332), that belongs to the amidohydrolase superfamily. NagA enzymes catalyze the deacetylation of -acetylglucosamine-6-phosphate (GlcNAc6P) to glucosamine-6-phosphate (GlcN6P). NagA is a potential anti-tubercular drug target because it represents the key enzymatic step in the generation of essential amino-sugar precursors required for cell wall biosynthesis and also influences recycling of cell wall peptidoglycan fragments. Here, we report the structural and functional characterization of NagA from (MSNagA) and (MMNagA), close relatives of Using a combination of X-ray crystallography, site-directed mutagenesis, and biochemical and biophysical assays, we show that these mycobacterial NagA enzymes are selective for GlcNAc6P. Site-directed mutagenesis studies revealed crucial roles of conserved residues in the active site that underpin stereo-selective recognition, binding, and catalysis of substrates. Moreover, we report the crystal structure of MSNagA in both ligand-free form and in complex with the GlcNAc6P substrate at 2.6 Å and 2.0 Å resolutions, respectively. The GlcNAc6P-complex structure disclosed the precise mode of GlcNAc6P binding and the structural framework of the active site, including two divalent metals located in the α/β binuclear site. Furthermore, we observed a cysteine residue located on a flexible loop region that occludes the active site. This cysteine is unique to mycobacteria and may represent a unique subsite for targeting mycobacterial NagA enzymes. Our results provide critical insights into the structural and mechanistic properties of mycobacterial NagA enzymes having an essential role in amino-sugar and nucleotide metabolism in mycobacteria

Physical activity and pre-diabetes—an unacknowledged mid-life crisis: findings from NHANES 2003–2006

The prevalence of pre-diabetes (PD) among US adults has increased substantially over the past two decades. By current estimates, over 34% of US adults fall in the PD category, 84% of whom meet the American Diabetes Association’s criteria for impaired fasting glucose (IFG). Low physical activity (PA) and/or sedentary behavior are key drivers of hyperglycemia. We examined the relationship between PD and objectively measured PA in NHANES 2003–2006 of 20,470 individuals, including 7,501 individuals between 20 and 65 yrs.We excluded all participants without IFG measures or adequate accelerometry data (final N = 1,317). Participants were identified as PD if FPG was 100–125 mg/dL (5.6–6.9 mmol/L). Moderate and vigorous PA in minutes/day individuals were summed to create the exposure variable “moderate-vigorous PA” (MVPA). The analysis sample included 884 normoglycemic persons and 433 with PD. There were significantly fewer PD subjects in the middle (30.3%) and highest (24.6%) tertiles of PA compared to the lowest tertile (35.5%). After adjusting for BMI, participants were 0.77 times as likely to be PD if they were in the highest tertile compared to the lowest PA tertile (p < 0.001). However, these results were no longer significant when age and BMI were held constant. Univariate analysis revealed that physical activity was associated with decreased fasting glucose of 0.5 mg/dL per minute of MVPA, but multivariate analysis adjusting for age and BMI was not significant. Overall, our data suggest a negative association between measures of PA and the prevalence of PD in middle-aged US adults independent of adiposity, but with significant confounding influence from measures of BMI and age

The combined effects of acidification and acute warming on the embryos of Pacific herring (Clupea pallasii)

Anthropogenic climate change is projected to affect marine ecosystems by challenging the environmental tolerance of individuals. Marine fishes may be particularly vulnerable to emergent climate stressors during early life stages. Here we focus on embryos of Pacific herring (Clupea pallasii), an important forage fish species widely distributed across the North Pacific. Embryos were reared under a range of temperatures (10-16°C) crossed with two pCO2 levels (600 and 2000 μatm) to investigate effects on metabolism and survival. We further tested how elevated pCO2 affects critical thermal tolerance (CTmax) by challenging embryos to short-term temperature fluctuations. Experiments were repeated on embryos collected from winter and spring spawning populations to determine if spawning phenology corresponds with different limits of environmental tolerance in offspring. We found that embryos could withstand acute exposure to 20°C regardless of spawning population or incubation treatment, but that survival was greatly reduced after 2-3 hours at 25°C. We found that pCO2 had limited effects on CTmax. The survival of embryos reared under chronically warm conditions (12°, 14°, or 16°C) was significantly lower relative to 10°C treatments in both populations. Oxygen consumption rates (MO2) were also higher at elevated temperatures and pCO2 levels. However, heart contraction measurements made 48 hours after CTmax exposure revealed a greater increase in heart rate in embryos reared at 10°C compared to 16°C, suggesting acclimation at higher incubation temperatures. Our results indicate that Pacific herring are generally tolerant of pCO2 but are vulnerable to acute temperature stress. Importantly, spring-spawning embryos did not clearly exhibit a higher tolerance to heat stress compared to winter offspring

Motor development in infancy and spine shape in early old age: findings from a British birth cohort study

Spine shape changes dramatically in early life, influenced by attainment of developmental milestones such as independent walking. Whether these associations persist across life is unknown. Therefore, we investigated associations between developmental milestones and spine shape, as determined using statistical shape models (SSMs) of lumbar spine from DXA scans in 1327 individuals (688 female) at 60‐64y in the MRC National Survey of Health and Development. Lumbar lordosis angle (L4 inferior endplate to T12 superior endplate) was measured using the two‐line Cobb method. In analyses adjusted for sex, height, lean and fat mass, socioeconomic position and birthweight, later walking age was associated with greater lordosis described by SSM1 (regression coefficient 0.023, 95%CI 0.000‐0.047, p=0.05) and direct angle measurement. Modest associations between walking age and less variation in anterior‐posterior vertebral size caudally (SSM6) were also observed (0.021, 95%CI ‐0.002‐0.044, p=0.07). Sex interactions showed that later walking was associated with larger relative vertebral anterior‐posterior dimensions in men (SSM3; ‐0.043, 95%CI ‐0.075‐0.01, p=0.01) but not women (0.018, 95%CI ‐0.0007‐0.043, p=0.17). Similar associations were observed between age at independent standing and SSMs but there was little evidence of association between sitting age and spine shape. Unadjusted associations between walking age and SSMs 1 and 6 remained similar after adjustment for potential confounders and mediators. This suggests that these associations may be explained by altered mechanical loading of the spine during childhood growth, although other factors could contribute. Early life motor development, particularly walking, may have a lasting effect on features of spine morphology with clinical significance

A novel syndrome of paediatric cataract, dysmorphism, ectodermal features, and developmental delay in Australian Aboriginal family maps to 1p35.3-p36.32

Background: A novel phenotype consisting of cataract, mental retardation, erythematous skin rash and facial dysmorphism was recently described in an extended pedigree of Australian Aboriginal descent. Large scale chromosomal re-arrangements had previously been ruled out. We have conducted a genome-wide scan to map the linkage region in this family.Methods: Genome-wide linkage analysis using Single Nucleotide Polymorphism (SNP) markers on the Affymetrix 10K SNP array was conducted and analysed using MERLIN. Three positional candidate genes (ZBTB17, EPHA2 and EPHB2) were sequenced to screen for segregating mutations. Results: Under a fully penetrant, dominant model, the locus for this unique phenotype was mapped to chromosome 1p35.3-p36.32 with a maximum LOD score of 2.41. The critical region spans 48.7 cM between markers rs966321 and rs1441834 and encompasses 527 transcripts from 364 annotated genes. No coding mutations were identified in three positional candidate genes EPHA2, EPHB2 or ZBTB17. The region overlaps with a previously reported region for Volkmann cataract and the phenotype has similarity to that reported for 1p36 monosomy. Conclusions: The gene for this syndrome is located in a 25.6 Mb region on 1p35.3-p36.32. The known cataract gene in this region (EPHA2) does not harbour mutations in this family, suggesting that at least one additional gene for cataract is present in this region.Kathryn Hattersley, Kate J Laurie, Jan E Liebelt, Jozef Gecz, Shane R Durkin, Jamie E Craig and Kathryn P Burdo

Quantifying Kinematic Substructure in the Milky Way's Stellar Halo

We present and analyze the positions, distances, and radial velocities for over 4000 blue horizontal-branch (BHB) stars in the Milky Way's halo, drawn from SDSS DR8. We search for position-velocity substructure in these data, a signature of the hierarchical assembly of the stellar halo. Using a cumulative "close pair distribution" (CPD) as a statistic in the 4-dimensional space of sky position, distance, and velocity, we quantify the presence of position-velocity substructure at high statistical significance among the BHB stars: pairs of BHB stars that are close in position on the sky tend to have more similar distances and radial velocities compared to a random sampling of these overall distributions. We make analogous mock-observations of 11 numerical halo formation simulations, in which the stellar halo is entirely composed of disrupted satellite debris, and find a level of substructure comparable to that seen in the actually observed BHB star sample. This result quantitatively confirms the hierarchical build-up of the stellar halo through a signature in phase (position-velocity) space. In detail, the structure present in the BHB stars is somewhat less prominent than that seen in most simulated halos, quite possibly because BHB stars represent an older sub-population. BHB stars located beyond 20 kpc from the Galactic center exhibit stronger substructure than at $\rm r_{gc} < 20$ kpc.Comment: 29 page, 10 figures, 1 table; accepted by APJ; for related article by another group see arXiv:1011.192

"I just keep thinking that I don't want to rely on people." a qualitative study of how people living with dementia achieve and maintain independence at home: stakeholder perspectives

YesBACKGROUND: Most people living with dementia want to remain in their own homes, supported by family and paid carers. Care at home often breaks down, necessitating transition to a care home and existing interventions are limited. To inform the development of psychosocial interventions to enable people with dementia to live well for longer at home, we qualitatively explored the views of people living with dementia, family carers and health and social care professionals, on how to achieve and maintain independence at home and what impedes this. METHODS: We conducted an inductive thematic analysis of qualitative interviews with 11 people living with dementia, 19 professionals and 22 family carers in England. RESULTS: We identified four overarching themes: being in a safe and familiar environment, enabling not disabling care, maintaining relationships and community connectedness, and getting the right support. For people living with dementia, the realities of staying active were complex: there was a tension between accepting support that enabled independence and a feeling that in doing so they were accepting dependency. Their and professionals' accounts prioritised autonomy and 'living well with dementia', while family carers prioritised avoiding harm. Professionals promoted positive risk-taking and facilitating independence, whereas family carers often felt they were left holding this risk. DISCUSSION: Psychosocial interventions must accommodate tensions between positive risk-taking and avoiding harm, facilitating autonomy and providing support. They should be adaptive and collaborative, combining self-management with flexible support. Compassionate implementation of rights-based dementia care must consider the emotional burden for family carers of supporting someone to live positively with risk.This work was supported by the Alzheimer’s Society (UK) and was carried out within the UCL Alzheimer’s Society Centre of Excellence for Independence at home, NIDUS (New Interventions in Dementia Study) programme (Alzheimer’s Society Centre of Excellence grant 330). This project is also part-funded funded by The National Institute for Health Research Applied Research Collaboration North West Coast (ARC NWC)