Comparative analysis of dna methylation profiles in peripheral blood leukocytes versus lymphoblastoid cell lines

Previous reports have shown that DNA methylation profiles within primary human malignant tissues are altered when these cells are transformed into cancer cell lines. However, it is unclear if similar differences in DNA methylation profiles exist between DNA derived from peripheral blood leukocytes (PBLs) and corresponding Epstein-Barr Virus transformed lymphoblastoid cell lines (LCLs). To assess the utility of LCLs as a resource for methylation studies we have compared DNA methylation profiles in promoter and 5' regions of 318 genes in PBL and LCL sample pairs from patients with type 1 diabetes with or without nephropathy. We identified a total of 27 (similar to 8%) genes that revealed different DNA methylation profiles in PBL compared with LCL-derived DNA samples. In conclusion, although the profiles for most promoter regions were similar between PBL-LCL pairs, our results indicate that LCL-derived DNA may not be suitable for DNA methylation studies at least in diabetic nephropathy.

Vasoactivity of AG014699, a clinically active small molecule inhibitor of poly(ADP-ribose) polymerase: a contributory factor to chemopotentiation in vivo?

Purpose: Poly(ADP-ribose) polymerase (PARP) plays an important role in DNA repair, and PARP inhibitors can enhance the activity of DNA-damaging agents in vitro and in vivo. AG014699 is a potent PARP inhibitor in phase II clinical development. However, the range of therapeutics with which AG014699 could interact via a DNA-repair based mechanism is limited. We aimed to investigate a novel, vascular-based activity of AG014699, underlying in vivo chemosensitization, which could widen its clinical application.

Experimental Design: Temozolomide response was analyzed in vitro and in vivo. Vessel dynamics were monitored using “mismatch” following the administration of perfusion markers and real-time analysis of fluorescently labeled albumin uptake in to tumors established in dorsal window chambers. Further mechanistic investigations used ex vivo assays of vascular smooth muscle relaxation, gut motility, and myosin light chain kinase (MLCK) inhibition.

Results: AG014699 failed to sensitize SW620 cells to temozolomide in vitro but induced pronounced enhancement in vivo. AG014699 (1 mg/kg) improved tumor perfusion comparably with the control agents nicotinamide (1 g/kg) and AG14361 (forerunner to AG014699; 10 mg/kg). AG014699 and AG14361 relaxed preconstricted vascular smooth muscle more potently than the standard agent, hydralazine, with no impact on gut motility. AG014699 inhibited MLCK at concentrations that relaxed isolated arteries, whereas AG14361 had no effect.

Conclusion: Increased vessel perfusion elicited by AG014699 could increase tumor drug accumulation and therapeutic response. Vasoactive concentrations of AG014699 do not cause detrimental side effects to gut motility and may increase the range of therapeutics with which AG014699 could be combined with for clinical benefit.

Facile synthesis of two diastereomeric indolizidines corresponding to the postulated structure of alkaloid 5,9E-259B from a Bufonid toad (Melanophryniscus)

A short synthesis of the postulated structure for indolizidine alkaloid 259B with the hydrogens at C5 and C9 entgegen has been achieved with complete control of stereochemistry at C5. Both diastereoisomers at C8 were obtained, but neither proved to be the natural product. The comparison of the mass and FTIR spectral properties of the synthetic compounds to those of the natural material strongly suggest that the gross structure is correct and that the difference may be a branch in the C5 alkyl side-chain. The GC-retention times of the two synthetic compounds were markedly longer than that of the natural 5,9E-259B.

Co-regulation of Gremlin and Notch signalling in diabetic nephropathy

Diabetic nephropathy is currently the leading cause of end-stage renal disease worldwide, and occurs in approximately one third of all diabetic patients. The molecular pathogenesis of diabetic nephropathy has not been fully characterized and novel mediators and drivers of the disease are still being described. Previous data from our laboratory has identified the developmentally regulated gene Gremlin as a novel target implicated in diabetic nephropathy in vitro and in vivo. We used bioinformatic analysis to examine whether Gremlin gene sequence and structure could be used to identify other genes implicated in diabetic nephropathy. The Notch ligand Jagged1 and its downstream effector, hairy enhancer of split-1 (Hes1), were identified as genes with significant similarity to Gremlin in terms of promoter structure and predicted microRNA binding elements. This led us to discover that transforming growth factor-beta (TGFß1), a primary driver of cellular changes in the kidney during nephropathy, increased Gremlin, Jagged1 and Hes1 expression in human kidney epithelial cells. Elevated levels of Gremlin, Jagged1 and Hes1 were also detected in extracts from renal biopsies from diabetic nephropathy patients, but not in control living donors. In situ hybridization identified specific upregulation and co-expression of Gremlin, Jagged1 and Hes1 in the same tubuli of kidneys from diabetic nephropathy patients, but not controls. Finally, Notch pathway gene clustering showed that samples from diabetic nephropathy patients grouped together, distinct from both control living donors and patients with minimal change disease. Together, these data suggest that Notch pathway gene expression is elevated in diabetic nephropathy, co-incident with Gremlin, and may contribute to the pathogenesis of this disease.

From the "pólis" to the modern state: an approach to the problem of politics in Heidegger

This paper starts off asking whether a strictly political approach may be deduced based on Martin Heidegger’ ontological analyses of modernity. His interpretation of the Greek phenomenon of the polis is discussed along with the distinction established therein between this form of community and the modern state, founded according to Heidegger on the metaphysical essence of modernity. To clarify this question regard is had to the proclamation of values observed by Heidegger in the different forms of state organization arising in the age of technical consummation of metaphysics. In this connection, his vision of nihilism is studied and a hypothesis is finally offered as to the form of state that would be consistent with a renunciation of the values required, in his view, by the manifestation of the entity in modernity as a wholly producible object.

FKBPL regulates estrogen receptor signaling and determines response to endocrine therapy

The HSP90 chaperone and immunophilin FKBPL is an estrogen-responsive gene that interacts with estogen receptor a (ERa) and regulates its levels. In this study, we explored the effects of FKBPL on breast cancer proliferation. Breast cancer cells stably overexpressing FKBPL became dependent on estrogen for their growth and were dramatically more sensitive to the antiestrogens tamoxifen and fulvestrant, whereas FKBPL knockdown reverses this phenotype. FKBPL knockdown also decreased the levels of the cell cycle inhibitor p21WAF1 and increased ERa phosphorylation on Ser118 in response to 17ß-estradiol and tamoxifen. In support of the likelihood that these effects explained FKBPL-mediated cell growth inhibition and sensitivity to endocrine therapies, FKBPL expression was correlated with increased overall survival and distant metastasis-free survival in breast cancer patients. Our findings suggest that FKBPL may have prognostic value based on its impact on tumor proliferative capacity and sensitivity to endocrine therapies, which improve outcome.

The Role of Process Parameters in Determining Wall Thickness Distribution in Plug-Assisted Thermoforming

This article describes the results of a comprehensive investigation to determine the link between process parameters and observed wall thickness output for the plug-assisted thermoforming process. The overall objective of the work was to systematically investigate the process parameters that may be adjusted during production to control the wall thickness distribution of parts manufactured by plug-assisted thermoforming. The parameters investigated were the sheet temperature, plug temperature, plug speed, plug displacement, plug shape, and air pressure. As well as quantifying the effects of each parameter on the wall thickness distribution, a further aim of the work was to improve the understanding of the physical mechanisms of deformation of the sheet during the different stages of the process. The process parameters shown to have the greatest effect on experimentally determined wall thickness distribution were the plug displacement, sheet temperature, plug temperature, and plug shape. It is proposed that during the plug-assisted thermoforming of polystyrene the temperature dependent friction between the plug and sheet surface was the most important factor in determining product wall thickness distribution, whereas heat transfer was shown to play a less important role. POLYM. ENG. SCI., 2010. © 2010 Society of Plastics Engineers

Intraguild predation may explain an amphipod replacement: evidence from laboratory populations

In a laboratory experiment that permitted both observations of the behaviour of individuals and the monitoring of small populations, the role of 'intraguild predation' in the elimination of the freshwater amphipod Gammarus duebeni celticus by the introduced G. pulex was examined. Over 18 weeks, deaths in single and mixed species replicates were monitored. Rates of 'mortality' (deaths not due to cannibalism or predation) did not differ between the species. Gammarus cl. celticus, however, was more cannibalistic than G. pulex and, in both species, males were more often cannibalized than females. In mixed species replicates, the mean proportions of animals preyed upon did not differ among replicates with differing starting proportions of the two species, nor was there a difference between the sexes in numbers preyed upon. G. pulex, however, preyed more frequently on G. d celticus than vice versa, and this became more pronounced over time. In 87% of mixed species replicates, G. pulex eliminated G. d. celticus. The results support the proposition that intraguild predation may be the primary mechanism whereby G. pulex rapidly replaces G. d. celticus in freshwater. Integrating behavioural observations with population level monitoring may thus link pattern and process in behaviour and ecology.

Parasitism may enhance rather than reduce the predatory impact of an invader

Invasive species can have profound impacts on communities and it is increasingly recognized that such effects may be mediated by parasitism. The 'enemy release' hypothesis posits that invaders may be successful and have high impacts owing to escape from parasitism. Alternatively, we hypothesize that parasites may increase host feeding rates and hence parasitized invaders may have increased community impacts. Here, we investigate the influence of parasitism on the predatory impact of the invasive freshwater amphipod Gammarus pulex. Up to 70 per cent of individuals are infected with the acanthoce- phalan parasite Echinorhynchus truttae, but parasitized individuals were no different in body condition to those unparasitized. Parasitized individuals consumed significantly more prey (Asellus aquaticus; Isopoda) than did unparasitized individuals. Both parasitized and unparasitized individuals displayed Type-II functional responses (FRs), with the FR for parasitized individuals rising more steeply, with a higher asymptote, compared with unparasi- tized individuals. While the parasite reduced the fitness of individual females, we predict a minor effect on population recruitment because of low parasite prevalence in the peak reproductive period. The parasite thus has a large per capita effect on predatory rate but a low population fitness effect, and thus may enhance rather than reduce the impact of this invader.

Homogeneous habitat can meet the discrete and varied resource requirements of hares but may set an ecological trap

In common with other farmland species, hares (Lepus spp.) are in widespread decline in agricultural landscapes due to agricultural intensification and habitat loss. We examined the importance of habitat heterogeneity to the Irish hare (Lepus timidus hibernicus) in a pastoral landscape. We used radio-tracking during nocturnal active and diurnal inactive periods throughout one year. In autumn, winter and spring, hares occupied a heterogeneous combination of improved grassland, providing food, and Juncus-dominated rough pasture, providing refuge. In summer, hares significantly increased their use of improved grassland. This homogeneous habitat can fulfil the discrete and varied resource requirements of hares for feeding and shelter at certain times of year. However, improved grassland may be a risky habitat for hares as silage harvesting occurs during their peak birthing period of late spring and early summer. We therefore posit the existence of a putative ecological trap inherent to a homogeneous habitat of perceived high value that satisfies the hares' habitat requirements but which presents risks at a critical time of year. To test this hypothesis in relation to hare populations, work is required to provide data on differential leveret mortality between habitat types.