Proteomic profiling of human retinal pigment epithelium exposed to an advanced glycation-modified substrate

Purpose The retinal pigment epithelium (RPE) and underlying Bruch’s membrane undergo significant modulation during ageing. Progressive, age-related modifications of lipids and proteins by advanced glycation end products (AGEs) at this cell–substrate interface have been implicated in RPE dysfunction and the progression to age-related macular degeneration (AMD). The pathogenic nature of these adducts in Bruch’s membrane and their influence on the overlying RPE remains unclear. This study aimed to identify alterations in RPE protein expression in cells exposed to AGE-modified basement membrane (AGE-BM), to determine how this “aged” substrate impacts RPE function and to map the localisation of identified proteins in ageing retina. Methods Confluent ARPE-19 monolayers were cultured on AGE-BM and native, non-modified BM (BM). Following 28-day incubation, the proteome was profiled using 2-dimensional gel electrophoresis (2D), densitometry and image analysis was employed to map proteins of interest that were identified by electrospray ionisation mass spectrometry (ESI MS/MS). Immunocytochemistry was employed to localise identified proteins in ARPE-19 monolayers cultured on unmodified and AGE-BM and to analyze aged human retina. Results Image analysis detected altered protein spot densities between treatment groups, and proteins of interest were identified by LC ESI MS/MS which included heat-shock proteins, cytoskeletal and metabolic regulators. Immunocytochemistry revealed deubiquitinating enzyme ubiquitin carboxyterminal hydrolase-1 (UCH-L1), which was upregulated in AGE-exposed RPE and was also localised to RPE in human retinal sections. Conclusions This study has demonstrated that AGE-modification of basement membrane alters the RPE proteome. Many proteins are changed in this ageing model, including UCHL-1, which could impact upon RPE degradative capacity. Accumulation of AGEs at Bruch”s membrane could play a significant role in age-related dysfunction of the RPE.

Synthesis and SAR of novel, 4-(phenylsulfamoyl)phenylacetamide mGlu(4) positive allosteric modulators (PAMs) identified by functional high-throughput screening (HTS)

Herein we disclose the synthesis and SAR of a series of 4-(phenylsulfamoyl)phenylacetamide compounds as mGlu(4) positive allosteric modulators (PAMs) that were identified via a functional HTS. An iterative parallel approach to these compounds culminated in the discovery of VU0364439 (11) which represents the most potent (19.8 nM) mGlu(4) PAM reported to date. (C) 2010 Elsevier Ltd. All rights reserved.

Analysis of pharmacists attitudes toward a distance learning initiative on health screening

A questionnaire was developed to investigate pharmacists' attitudes to distance learning (DL) as a vehicle for continuing education (CE). It was included in each of a two part DL course on Health Screening. Part One was mailed to all community pharmacists in England (16,400) and returns were received from 1487. The questionnaire in Part Two was returned by 436 pharmacists. Attitude statements were scored using a five-point Likert scale. The mean response to all attitude statements was positive. Participants were significantly more satisfied than non-participants with DL in general and the DL course studied (P less than or equal to 0.05). Over 80 percent of respondents completing the course found DL to be enjoyable and more suitable than other CE methods. More females and less males than expected (based on registration statistics) requested (P less than or equal to 0.001) and completed the course (P less than or equal to 0.001). Pharmacists of all ages participated, although those recently qualified showed greater interest.

Uptake of breast screening is influenced by current religion and religion of upbringing

Research has shown that individuals with a current religious affiliation are more likely to use preventive health services. The aim of this study was to determine whether breast screening uptake in Northern Ireland is higher amongst women with a current affiliation to an organised religion and, for those with no current affiliation, to examine whether their religion of upbringing is associated with uptake of breast screening. The Northern Ireland Longitudinal Study (NILS) was used to link Census and national breast screening data for 37,211 women invited for routine breast screening between 2001 and 2004. Current religious affiliation, religion of upbringing and other demographic and socio-economic characteristics were as defined on the Census form. Multivariate logistic regression was used to determine the relationship between religion affiliation and attendance. Uptake of breast screening is about 25% lower for those without a current religious affiliation. There are modest differences between Catholics and Protestants, with the latter about 11% more likely to attend for screening. For those with no current religion, the religion of upbringing appears to positively influence attendance rates. These differences remain after adjustment for all of the socio-demographic and socio-economic factors that have been shown to influence uptake rates of breast screening in the UK to date. Record linkage is an efficient way to examine equity across demographic characteristics that are not routinely available. The lower uptake amongst those with no religious affiliation may mean that screening services may find it difficult to maintain or improve uptake rate in an increasingly secularised society.