In silico analysis and DHPLC screening strategy identifies novel apoptotic gene targets of aberrant promoter hypermethylation in prostate cancer

BACKGROUND: Aberrant DNA methylation has been implicated as a key survival mechanism in cancer, whereby promoter hypermethylation silences genes essential for many cellular processes including apoptosis. Limited data is available on the methylation profile of apoptotic genes in prostate cancer (CaP). The aim of this study was to profile methylation of apoptotic-related genes in CaP using denaturing high performance liquid chromatography (DHPLC).

METHODS: Based on an in silico selection process, 13 genes were screened for methylation in CaP cell lines using DHPLC. Quantitative methylation specific PCR was employed to determine methylation levels in prostate tissue specimens (n = 135), representing tumor, histologically benign prostate, high-grade prostatic intraepithelial neoplasia and benign prostatic hyperplasia. Gene expression was measured by QRT-PCR in cell lines and tissue specimens.

RESULTS: The promoters of BIK, BNIP3, cFLIP, TMS1, DCR1, DCR2, and CDKN2A appeared fully or partially methylated in a number of malignant cell lines. This is the first report of aberrant methylation of BIK, BNIP3, and cFLIP in CaP. Quantitative methylation analysis in prostate tissues identified 5 genes (BNIP3, CDKN2A, DCR1, DCR2 and TMS1) which were frequently methylated in tumors but were unmethylated in 100% of benign tissues. Furthermore, 69% of tumors were methylated in at least one of the five-gene panel. In the case of all genes, except BNIP3, promoter hypermethylation was associated with concurrent downregulation of gene expression.

CONCLUSION: Future examination of this "CaP apoptotic methylation signature" in a larger cohort of patients is justified to further evaluate its value as a diagnostic and prognostic marker.

Discovery of DNA hypermethylation using a DHPLC screening strategy

Promoter hypermethylation is recognized as a hallmark of human cancer, in addition to conventional mechanisms of gene inactivation. As such, many new technologies have been developed over the past two decades to uncover novel targets of methylation and decipher complex epigenetic patterns. However, many of these are either labor intensive or provide limited data, confined to oligonucleotide hybridization sequences or enzyme cleavage sites and cannot be easily applied to screening large sets of sequences or samples. We present an application of denaturing high performance liquid chromatography (DHPLC), which relies on bisulfite modification of genomic DNA, for methylation screening. We validated DHPLC as a methylation screening tool using GSTP1, a well known target of methylation in prostate cancer. We developed an in silico approach to identify potential targets of promoter hypermethylation in prostate cancer. Using DHPLC, we screened two of these targets LGALS3 and SMAD4 for methylation. We show that DHPLC has an application as a fast, sensitive, quantitative and cost effective method for screening novel targets or DNA samples for DNA methylation.

An aggregated fridge-freezer peak shaving and valley filling control strategy for enhanced grid operations

The need for fast response demand side participation (DSP) has never been greater due to increased wind power penetration. White domestic goods suppliers are currently developing a `smart' chip for a range of domestic appliances (e.g. refrigeration units, tumble dryers and storage heaters) to support the home as a DSP unit in future power systems. This paper presents an aggregated population-based model of a single compressor fridge-freezer. Two scenarios (i.e. energy efficiency class and size) for valley filling and peak shaving are examined to quantify and value DSP savings in 2020. The analysis shows potential peak reductions of 40 MW to 55 MW are achievable in the Single wholesale Electricity Market of Ireland (i.e. the test system), and valley demand increases of up to 30 MW. The study also shows the importance of the control strategy start time and the staggering of the devices to obtain the desired filling or shaving effect.

Failure to produce an anti-poverty strategy: The Committee on the Administration’s Application [2015] NIQB 59, June 30, 2015

This is a note on the Northern Ireland High Court decision of 30 June 2015 that the Northern Ireland Executive had acted unlawfully in failing to fulfil its statutory duty to adopt a strategy setting out proposals for tackling poverty, social exclusion and patterns of deprivation based on objective need.

Towards the 2021 UK Census imputation strategy: Response mode as a matching variable in a donor-based approach?

Since July 2014, the Office for National Statistics has committed to a predominantly online 2021 UK Census. Item-level imputation will play an important role in adjusting the 2021 Census database. Research indicates that the internet may yield cleaner data than paper based capture and attract people with particular characteristics. Here, we provide preliminary results from research directed at understanding how we might manage these features in a 2021 UK Census imputation strategy. Our findings suggest that if using a donor-based imputation method, it may need to consider including response mode as a matching variable in the underlying imputation model.

An Association of Cancer Physicians' strategy for improving services and outcomes for cancer patients

The Association of Cancer Physicians in the United Kingdom has developed a strategy to improve outcomes for cancer patients and identified the goals and commitments of the Association and its members.

A comprehensive strategy to detect the fraudulent adulteration of herbs: The oregano approach

Fraud in the global food supply chain is becoming increasingly common due to the huge profits associated with this type of criminal activity. Food commodities and ingredients that are expensive and are part of complex supply chains are particularly vulnerable. Both herbs and spices fit these criteria perfectly and yet strategies to detect fraudulent adulteration are still far from robust. An FT-IR screening method coupled to data analysis using chemometrics and a second method using LC-HRMS were developed, with the latter detecting commonly used adulterants by biomarker identification. The two tier testing strategy was applied to 78 samples obtained from a variety of retail and on-line sources. There was 100% agreement between the two tests that over 24% of all samples tested had some form of adulterants present. The innovative strategy devised could potentially be used for testing the global supply chains for fraud in many different forms of herbs.

Mindfulness - As a coping strategy

Mindfulness has been described as “paying attention in a particular way: on purpose, in the present moment, and nonjudgmentally.” (Kabat-Zinn, 1994, p. 4). It is a technique where one focuses on the present, gradually letting go of thoughts about the past or the future. Mindfulness is becoming more popular as a technique to help people manage stress. Research suggests, for example, that individuals who have higher levels of mindfulness have increased performance in attention and cognitive flexibility (Moore & Malinowski, 2009); report higher levels of relationship satisfaction (Kozlowski, 2013), and lower levels of perceived stress (Roeser et al., 2013). As a therapeutic technique mindfulness has been shown to be effective through, for example, Mindfulness Based Cognitive Therapy and Mindfulness-Based Stress Reduction (Nevanper, 2012). Aims of Research The argument offered here is that mindfulness is likely to act in the same way as other types of coping i.e. that it is not a ‘silver bullet’ and that it is likely to be a preferred strategy used by some and not others. The aim of this research therefore is to compare the impact of mindfulness compared to other types of coping on well-being - operationalised as happiness, selfcompassion and stress.

An Evaluation of the Dual Diagnosis (substance use and mental health )Strategy in the South Eastern Trust Workers and Service Users Perceptions of Dual Diagnosis Issues

In the UK it is estimated that over 33% of psychiatric patients with enduring mental illness have a substance misuse problem, whilst over 50 % of clients currently accessing drug and alcohol services have a mental health problem. Between 2003 and 2013 in Northern Ireland, there were 741 recorded suicides by patients who were in contact with mental health services. Of this number, 68% (n=501) had a history of either alcohol or drug misuse or both, resulting in an average of 46 patient suicides per year associated with dual diagnosis (University of Manchester 2015).
The current evaluation examined staff attitudes towards working with dual diagnosis (co-existing difficulties) issues, staff confidence in working with clients with dual diagnosis, workers’ perceptions of the South Eastern dual diagnosis strategy and service user perspectives of dual diagnosis service provision.
The purpose of the evaluation was to provide evidence regarding the following in accordance with the current dual diagnosis strategy;
Staff understanding of the concept of dual diagnosis,
Staff attitudes towards working with dual diagnosis,
Staff confidence in working with individuals, who present with dual diagnosis,
Service users’ perspectives of SE Trust provision for dual diagnosis.
Staff views on the South Eastern Trust Dual Diagnosis Strategy.