Using co-production to implement patient reported outcome measures in third sector organisations: a mixed methods study.

Background: Third sector organisations such as charities and community groups are using Patient Reported Outcome Measures (PROMs) at an aggregated service level to demonstrate their impact to commissioners to generate or retain funding. Despite this motivation, organisations can struggle with implementing PROMs. Previous studies have identified facilitators including organisations using an appropriate measure, co-producing the PROMs process with staff, and investing resources to support the use of measures.

Care planning for children with lower limb amputation.

This article explores the pathways of care for children who undergo lower limb amputation, from pre-surgery to rehabilitation. The consequences of surgery are manifold, including that children and their families must cope with life with a disability, effects on mobility, greater demands on metabolic reserve, disfigurement, pain and discomfort. Care can be divided into multidisciplinary pre-operative, post-operative, mid-term rehabilitation, including prosthetic limb casting and fitting, and long-term rehabilitation over many weeks and years.

Flexible feature-based inhibition in visual search mediates magnified impairments of selection: evidence from carry-over effects under dynamic preview-search conditions.

Evidence for inhibitory processes in visual search comes from studies using preview conditions, where responses to new targets are delayed if they carry a featural attribute belonging to the old distractor items that are currently being ignored-the negative carry-over effect (Braithwaite, Humphreys, & Hodsoll, 2003). We examined whether inhibition was applied in the same manner across different types of displays or whether the inhibitory weighting applied to different features varied with their utility for the search task. To test this, we present the first empirical investigation of negative carry-over effects under the ecologically valid conditions of dynamic visual search.

Visual search at isoluminance: evidence for enhanced color weighting in standard sub-set and preview-based visual search.

Isoluminant displays depend on responses from the parvocellular visual stream, known to code color information. We examined the influence of isoluminance on attentional guidance by color using two procedures: (i) color sub-set search (Egeth, Virzi, & Garbart, 1984) and (ii) preview search (Watson & Humphreys, 1997). We used displays that do not generate a sub-set search advantage with luminant stimuli.

Retinoid-induced histone deacetylation inhibits telomerase activity in estrogen receptor-negative breast cancer cells.

Background: Multiple mechanisms regulate cancer-associated telomerase activity at the level of human telomerase reverse transcriptase (hTERT) transcription which may serve as novel targets for anticancer approaches.

Materials And Methods: The effects of prolonged all-trans retinoic acid (ATRA) exposure on hTERT regulation in estrogen receptor-negative SK-BR-3 breast cancer cells were examined.

Results: ATRA had a profound effect on the morphology and proliferation rate of the SK-BR-3 cells.

Measuring the spread of spreading suppression: a time-course analysis of spreading suppression and its impact on attentional selection.

We report three experiments investigating the time course of spreading suppression in visual search using preview conditions. A novel color-change procedure was employed in which a target letter changed into a new (singleton) color at various intervals after the onset of the search display. Performance when the singleton was unique across both preview and search displays was compared with that when the singleton carried the color of the preview display.

Genistein depletes telomerase activity through cross-talk between genetic and epigenetic mechanisms.

Genistein, a natural isoflavone found in soybean products, has been reported to down-regulate telomerase activity and that this prevents cancer and contributes to the apoptosis of cancer cells. However, the precise molecular mechanism by which genistein represses telomerase is not clear. Here, we show that genistein inhibits the transcription of hTERT (human telomerase reverse transcriptase), the catalytic subunit of the human telomerase enzyme, in breast MCF10AT benign cells and MCF-7 cancer cells in a time- and dose-dependent manner.

Differential expression of epigenetic modulators during human embryonic stem cell differentiation.

Although the progression of aging and the diseases associated with it are extensively studied, little is known about the initiation of the aging process. Telomerase is down-regulated early in embryonic differentiation, thereby contributing to telomeric attrition and aging. The mechanisms underlying this inhibition remain elusive, but epigenetic studies in differentiating human embryonic stem (hES) cells could give clues about how and when DNA methylation and histone deacetylation work together to contribute to the inactivation of hTERT, the catalytic subunit of telomerase, at the onset of the aging process.

The Novel Retinoid, 9cUAB30, Inhibits Telomerase and Induces Apoptosis in HL60 Cells.

Telomerase, a ribonucleoprotein important to neoplastic immortality, is up-regulated in approximately 85% of cancers, including leukemias. In this study, 9cUAB30, a novel retinoic acid, resulted in differentiation of HL60 leukemia cells as indicated by morphologic changes characteristic of granulocytes. It also caused a down-regulation of hTERT gene expression and a decrease in telomerase activity.

Adenylyl cyclase alpha and cAMP signaling mediate Plasmodium sporozoite apical regulated exocytosis and hepatocyte infection.

Malaria starts with the infection of the liver of the host by Plasmodium sporozoites, the parasite form transmitted by infected mosquitoes. Sporozoites migrate through several hepatocytes by breaching their plasma membranes before finally infecting one with the formation of an internalization vacuole. Migration through host cells induces apical regulated exocytosis in sporozoites.

Retrovirus-mediated RNA interference. Targeting hTERT through stable expression of short-hairpin RNA.

RNA interference (RNAi) has recently emerged as a reliable tool for studying the effects of knocking down or ablating the expression of specific genes. It is hoped that progress made in the laboratory toward in vitro down regulation of gene expression may be carried over into the clinic for treatment of diseases in which the expression of a specific gene is associated with initiation or progression of that disease. Such is the case with telomerase, an exciting drug target that has been the focus of numerous investigations with a wide variety of inhibitors.

RNA interference using a plasmid construct expressing short-hairpin RNA.

RNA interference (RNAi) is one of the most commonly used procedures for gene targeting in today's cutting edge technology and has great potential for use in clinical therapy. Using a plasmid construct that exogenously expresses short-hairpin RNAs (shRNAs) targeting a desired gene transcript not only helps to study the downstream effects of a gene product but also offers an alternative to viral vectors for gene therapy. Using a plasmid vector to knockdown a gene allows for long-term and permanent gene knockdown, without the need to generate knockout genotypes.

hTERT knockdown in human embryonic kidney cells using double-stranded RNA.

The method of RNA interference (RNAi) is an easy means of knocking down a gene without having to generate knockout mutants, which may prove to be difficult and time consuming. RNAi is a naturally occurring process that involves targeting the mRNA of a gene by introducing RNAs that are complementary to the target mRNA. The foreign RNAs activate an endogenous enzyme, DICER, which degrades the target mRNA.

Methods of telomerase inhibition.

Telomerase is central to cellular immortality and is a key component of most cancer cells although this enzyme is rarely expressed to significant levels in normal cells. Therefore, the inhibition of telomerase has garnered considerable attention as a possible anticancer approach. Many of the methods applied to telomerase inhibition focus on either of the two major components of the ribonucleoprotein holoenzyme, that is, the telomerase reverse transcriptase (TERT) catalytic subunit or the telomerase RNA (TR) component.

Epigenetic regulation of telomerase in retinoid-induced differentiation of human leukemia cells.

Changes in the promoter methylation of hTERT, the gene that encodes telomerase, a ribonucleoprotein responsible for replacing telomeric repeats, have been demonstrated in differentiating cells where hTERT is inhibited, suggesting epigenetic regulation of hTERT. All-trans retinoic acid (ATRA) induces differentiation in human leukemia cells and has had significant clinical success treating promyelocytic leukemia in what is termed 'differentiation therapy'. It is thought that the inhibition of telomerase is a target of retinoids and is closely tied to the differentiated phenotype.

A method to study the expression of DNA methyltransferases in aging systems in vitro.

The methylation of CpG dinucleotides located in key protein binding sites within gene regulatory regions often leads to gene silencing. A mechanism of aging is proposed whereby an accumulation of methylation at gene regulatory sites contributes to cellular senescence. DNA methyltransferases (DNMTs) are enzymes that catalyze the transfer of a methyl moiety from S-adenosyl-L-methionine (SAM) to the cytosine of a CpG dinucleotide and are responsible for establishing and maintaining methylation patterns in the genome.

A method to detect DNA methyltransferase I gene transcription in vitro in aging systems.

Epigenetic alterations of DNA play key roles in determining gene structure and expression. Methylation of the 5-position of cytosine is thought to be the most common modification of the genome in mammals. Studies have generally shown that hypermethylation in gene regulatory regions is associated with inactivation and reduced transcription and that alteration in established methylation patterns during development can affect embryonic viability.

Aging cell culture: methods and observations.

Culturing and subcultivation of normal human diploid fibroblasts have advanced our understanding of the molecular events involved in aging. This progress is leading to the development of therapies that slow or ablate the adverse physiological and pathological changes associated with aging. It has been established that normal human diploid fibroblasts can proliferate in culture for only finite periods of time.

Epigenetic and genetic mechanisms contribute to telomerase inhibition by EGCG.

The ends of human chromosomes are protected from the degradation associated with cell division by 15-20 kb long segments of hexameric repeats of 5'-TTAGGG-3' termed telomeres. In normal cells telomeres lose up to 300 bp of DNA per cell division that ultimately leads to senescence; however, most cancer cells bypass this lifespan restriction through the expression of telomerase. hTERT, the catalytic subunit essential for the proper function of telomerase, has been shown to be expressed in approximately 90% of all cancers.

The low-toxicity 9-cis UAB30 novel retinoid down-regulates the DNA methyltransferases and has anti-telomerase activity in human breast cancer cells.

Retinoic acids and their derivatives potentiate anti-cancer effects in breast cancer cells. The aberrant expression of telomerase is critical to the continued proliferation of most cancer cells. Thus, telomerase is an attractive target for chemoprevention and treatment of breast cancer.

Evidence of extra-telomeric effects of hTERT and its regulation involving a feedback loop.

The human telomerase reverse transcriptase (hTERT) is the catalytic subunit of the enzyme telomerase which is responsible for telomeric maintenance and extension. Using RNA interference to knock down hTERT mRNA expression, we provide evidence that hTERT exerts extra-telomeric effects on the cell cycle and on its own regulatory proteins, specifically: p53 and p21. We tested our hypothesis that hTERT regulates its own expression through effects on upstream regulatory genes using transformed human embryonic kidney (HEK 293) cells, p53 and p16(INK4a) null human ovarian cancer SKOV-3 cells, and p53-null MDA-MB-157 human mammary cancer cells.