Evaluation of c-FLIP in castrate-resistant prostate cancer antagonizes therapeutic response to androgen receptor-targeted therapy

Purpose: To characterize the importance of cellular Fas-associated death domain (FADD)–like interleukin 1ß-converting enzyme (FLICE) inhibitory protein (c-FLIP), a key regulator of caspase-8 (FLICE)–promoted apoptosis, in modulating the response of prostate cancer cells to androgen receptor (AR)–targeted therapy.

Experimental Design: c-FLIP expression was characterized by immunohistochemical analysis of prostatectomy tissue. The functional importance of c-FLIP to survival and modulating response to bicalutamide was studied by molecular and pharmacologic interventions.

Results: c-FLIP expression was increased in high-grade prostatic intraepithelial neoplasia and prostate cancer tissue relative to normal prostate epithelium (P < 0.001). Maximal c-FLIP expression was detected in castrate-resistant prostate cancer (CRPC; P < 0.001). In vitro, silencing of c-FLIP induced spontaneous apoptosis and increased 22Rv1 and LNCaP cell sensitivity to bicalutamide, determined by flow cytometry, PARP cleavage, and caspase activity assays. The histone deacetylase inhibitors (HDACi), droxinostat and SAHA, also downregulated c-FLIP expression, induced caspase-8- and caspase-3/7–mediated apoptosis, and increased apoptosis in bicalutamide-treated cells. Conversely, the elevated expression of c-FLIP detected in the CRPC cell line VCaP underpinned their insensitivity to bicalutamide and SAHA in vitro. However, knockdown of c-FLIP induced spontaneous apoptosis in VCaP cells, indicating its relevance to cell survival and therapeutic resistance.

Conclusion: c-FLIP reduces the efficacy of AR-targeted therapy and maintains the viability of prostate cancer cells. A combination of HDACi with androgen deprivation therapy may be effective in early-stage disease, using c-FLIP expression as a predictive biomarker of sensitivity. Direct targeting of c-FLIP, however, may be relevant to enhance the response of existing and novel therapeutics in CRPC. Clin Cancer Res; 18(14); 3822–33.

Embedded C programming: a practical course introducing programmable microprocessors

This paper presents a new laboratory-based module for embedded systems teaching, which addresses the current lack of consideration for the link between hardware development, software implementation, course content and student evaluation in a laboratory environment. The course introduces second year undergraduate students to the interface between hardware and software and the programming of embedded devices; in this case, the PIC (originally peripheral interface controller, later rebranded programmable intelligent computer) microcontroller. A hardware development board designed for use in the laboratories of this module is presented. Through hands on laboratory experience, students are encouraged to engage with practical problem-solving exercises and develop programming skills across a broad range of scenarios.

Rheological and heat transfer behaviour of the ionic liquid, [C(4)mim][NTf2]

Systematic experiments have been carried out on the thermal and rheological behaviour of the ionic liquid, 1-butyl-3-methylimidazolium bis{(trifluoromethyl)sulfonyl} imide, [C(4)mim][NTf2], and, for the first time, on the forced convective heat transfer of an ionic liquid under the laminar flow conditions. The results show that the thermal conductivity of the ionic liquid is similar to 0.13 W m(-1) K-1, which is almost independent of temperature between 25 and 40 degrees C. Rheological measurements show that the [C(4)mim][NTf2] liquid is a Newtonian fluid with its shear viscosity decreasing with increasing temperature according to the exponential law over a temperature range of 20-90 degrees C. The convective heat transfer experiments demonstrate that the thermal entrance length of the ionic liquid is very large due to its high viscosity and low thermal conductivity. The convective heat transfer coefficient is observed to be much lower than that of distilled water under the same conditions. The convective heat transfer data are also found to fit well to the convectional Shah's equation under the conditions of this work. (C) 2007 Elsevier Inc. All rights reserved.

Reconstructing apology: David Cameron's Bloody Sunday apology in the press

While there is an acknowledgement in apology research that political apologies are highly mediated, the process of mediation itself has lacked scrutiny. This article suggests that the idea of reconstruction helps to understand how apologies are mediated and evaluated. David Cameron's apology for Bloody Sunday is examined to see how he constructs four aspects of apology: social actors, consequences, categorization, and reasons. The reconstruction of those aspects by British, Unionist, and Nationalist press along with reconstructions made by soldiers in an online forum are considered. Data analysis was informed by thematic analysis and discourse analysis which helped to explore key aspects of reconstruction and how elements of Cameron's apology are altered in subsequent mediated forms of the apology. These mediated reconstructions of the apology allowed their authors to evaluate the apology in different ways. Thus, in this article, it is suggested that the evaluation of the apology by different groups is preceded by a reconstruction of it in accordance with rhetorical goals. This illuminates the process of mediation and helps to understand divergent responses to political apologies.

Serum amyloid A-related inflammation is lowered by increased fruit and vegetable intake, while high-sensitive C-reactive protein, IL-6 and E-selectin remain unresponsive

The present study assessed whether increased fruit and vegetable (F&V) intake reduced the concentrations of the inflammatory marker serum amyloid A (SAA) in serum, HDL2 and HDL3 and whether the latter reduction influenced any of the functional properties of these HDL subfractions. The present study utilised samples from two previous studies: (1) the FAVRIT (Fruit and Vegetable Randomised Intervention Trial) study - hypertensive subjects (systolic blood pressure (BP) range 140-190 mmHg; diastolic BP range 90-110 mmHg) were randomised to receive a 1-, 3- or 6-portion F&V/d intervention for 8 weeks, and (2) the ADIT (Ageing and Dietary Intervention Trial) study - older subjects (65-85 years) were randomised to receive a 2- or 5-portion F&V/d intervention for 16 weeks. HDL2 and HDL3 were isolated by rapid ultracentrifugation. Measurements included the following: serum high-sensitive C-reactive protein (hsCRP) by an immunoturbidimetric assay; serum IL-6 and E-selectin and serum-, HDL2- and HDL3-SAA by ELISA procedures; serum-, HDL2- and HDL3-cholesterol ester transfer protein (CETP) activity by a fluorometric assay. Although the concentrations of hsCRP, IL-6 and E-selectin were unaffected by increasing F&V intake in both studies (P>0·05 for all comparisons), those of SAA in HDL3 decreased in the FAVRIT cohort (P= 0·049) and those in HDL2 and HDL3 decreased in the ADIT cohort (P= 0·035 and 0·032), which was accompanied by a decrease in the activity of CETP in HDL3 in the FAVRIT cohort (P= 0·010) and in HDL2 in the ADIT cohort (P= 0·030). These results indicate that SAA responds to increased F&V intake, while other inflammatory markers remain unresponsive, and this leads to changes in HDL2 and HDL3, which may influence their antiatherogenic potential. Overall, the present study provides tangible evidence of the effectiveness of increased F&V intake, which may be of use to health policy makers and the general public.

Linking watershed terrain and hydrology to soil chemical properties, microbial communities and impacts on soil organic C in a humid mid-latitude forested watershed

Understanding the response of humid mid-latitude forests to changes in precipitation, temperature, nutrient cycling, and disturbance is critical to improving our predictive understanding of changes in the surface-subsurface energy balance due to climate change. Mechanistic understanding of the effects of long-term and transient moisture conditions are needed to quantify
linkages between changing redox conditions, microbial activity, and soil mineral and nutrient interactions on C cycling and greenhouse gas releases. To illuminate relationships between the soil chemistry, microbial communities and organic C we established transects across hydraulic and topographic gradients in a small watershed with transient moisture conditions. Valley bottoms tend to be more frequently saturated than ridge tops and side slopes which generally are only saturated when shallow storm flow zones are active. Fifty shallow (~36”) soil cores were collected during timeframes representative of low CO2, soil winter conditions and high CO2, soil summer conditions. Cores were subdivided into 240 samples based on pedology and analyses of the geochemical (moisture content, metals, pH, Fe species, N, C, CEC, AEC) and microbial (16S rRNA gene
amplification with Illumina MiSeq sequencing) characteristics were conducted and correlated to watershed terrain and hydrology. To associate microbial metabolic activity with greenhouse gas emissions we installed 17 soil gas probes, collected gas samples for 16 months and analyzed them for CO2 and other fixed and greenhouse gasses. Parallel to the experimental efforts our data is being used to support hydrobiogeochemical process modeling by coupling the Community Land Model (CLM) with a subsurface process model (PFLOTRAN) to simulate processes and interactions from the molecular to watershed scales. Including above ground processes (biogeophysics, hydrology, and vegetation dynamics), CLM provides mechanistic water, energy, and organic matter inputs to the surface/subsurface models, in which coupled biogeochemical reaction
networks are used to improve the representation of below-ground processes. Preliminary results suggest that inclusion of above ground processes from CLM greatly improves the prediction of moisture response and water cycle at the watershed scale.