School of Biology & Environmental Science

Influence of Surface Groups on Poly(propylene imine) Dendrimers Antiprion Activity

2013-04-15T13:36:32Z

Influence of Surface Groups on Poly(propylene imine) Dendrimers Antiprion Activity McCarthy, James M.; Moreno, Beatriz Rasines; Filippini, Damien; Komber, Hartmut; Maly, Marek; Cernescu, Michaela; Brutschy, Bernhard; Appelhans, Dietmar; Rogers, Mark S. Prion diseases are characterized by the accumulation of PrP(Sc), an aberrantly folded isoform of the host protein PrP(C). Specific forms of synthetic molecules known as dendrimers are able to eliminate protease-resistant PrP(Sc) in both an intracellular and in vitro setting. The properties of a dendrimer which govern this ability are unknown. We addressed the issue by comparing the in vitro antiprion ability of numerous modified poly(propylene-imine) dendrimers, which varied in size, structure, charge, and surface group composition. Several of the modified dendrimers, including an anionic glycodendrimer, reduced the level of protease resistant PrP(Sc) in a prion strain-dependent manner. This led to the formulation of a new working model for dendrimer/prion interactions which proposes dendrimers eliminate PrP(Sc) by destabilizing the protein and rendering it susceptible to proteolysis. This ability is not dependent on any particular charge of dendrimer, but does require a high density of reactive surface groups.

Influence of surface functionality of poly(propylene imine) dendrimers on protease resistance and propagation of the scrapie prion protein

2013-04-15T12:10:27Z

Influence of surface functionality of poly(propylene imine) dendrimers on protease resistance and propagation of the scrapie prion protein Fischer, Marlies; Appelhans, Dietmar; Schwarz, Simona; Klajnert, Barbara; Bryszewska, Maria; Voit, Brigitte; Rogers, Mark S. Accumulation of PrP(Sc), an insoluble and protease-resistant pathogenic isoform of the cellular prion protein (PrP(C)), is a hallmark in prion diseases. Branched polyamines, including PPI (poly(propylene imine)) dendrimers, are able to remove protease resistant PrP(Sc) and abolish infectivity, offering possible applications for therapy. These dendrimer types are thought to act through their positively charged amino surface groups. In the present study, the molecular basis of the antiprion activity of dendrimers was further investigated, employing modified PPI dendrimers in which the positively charged amino surface groups were substituted with neutral carbohydrate units of maltose (mPPI) or maltotriose (m3PPI). Modification of surface groups greatly reduced the toxicity associated with unmodified PPI but did not abolish its antiprion activity, suggesting that the presence of cationic surface groups is not essential for dendrimer action. PPI and mPPI dendrimers of generation 5 were equally effective in reducing levels of protease-resistant PrP(Sc) (PrP(res)) in a dose- and time-dependent manner in ScN2a cells and in pre-existing aggregates in homogenates from infected brain. Solubility assays revealed that total levels of PrP(Sc) in scrapie-infected mouse neuroblastoma (ScN2a) cells were reduced by mPPI. Coupled with the known ability of polyamino dendrimers to render protease-resistant PrP(Sc) in pre-existing aggregates of PrP(Sc) susceptible to proteolysis, these findings strongly suggest that within infected cells dendrimers reduce total amounts of PrP(Sc) by mediating its denaturation and subsequent elimination.

Differentiating prion strains using dendrimers

2013-04-15T12:01:25Z

Differentiating prion strains using dendrimers McCarthy, James M.; Rasines, Beatriz; Appelhans, Dietmar; Rogers, Mark S. A panel of repetitively branched synthetic molecules known as dendrimers is used to identify and differentiate between different strains of the prion infectious agent, the protein-based pathogen responsible for prion disorders—a group of invariably fatal neurodegenerative diseases

Anti-prion drug mPPIg5 inhibits PrPC conversion to PrPSc

2013-04-15T11:54:11Z

Anti-prion drug mPPIg5 inhibits PrPC conversion to PrPSc McCarthy, James M.; Franke, Markus; Resenberger, Ulrike K.; Waldron, Sibeal; Simpson, Jeremy C.; Tatzelt, Jörg; Appelhans, Dietmar; Rogers, Mark S. Prion diseases, also known as transmissible spongiform encephalopathies, are a group of fatal neurodegenerative diseases that include scrapie in sheep, bovine spongiform encephalopathy (BSE) in cattle and Creutzfeldt-Jakob disease (CJD) in humans. The 'protein only hypothesis' advocates that PrPSc, an abnormal isoform of the cellular protein PrPC, is the main and possibly sole component of prion infectious agents. Currently, no effective therapy exists for these diseases at the symptomatic phase for either humans or animals, though a number of compounds have demonstrated the ability to eliminate PrPSc in cell culture models. Of particular interest are synthetic polymers known as dendrimers which possess the unique ability to eliminate PrPSc in both an intracellular and in vitro setting. The efficacy and mode of action of the novel anti-prion dendrimer mPPIg5 was investigated through the creation of a number of innovative bio-assays based upon the scrapie cell assay. These assays were used to demonstrate that mPPIg5 is a highly effective anti-prion drug which acts, at least in part, through the inhibition of PrPC to PrPSc conversion. Understanding how a drug works is a vital component in maximising its performance. By establishing the efficacy and method of action of mPPIg5, this study will help determine which drugs are most likely to enhance this effect and also aid the design of dendrimers with anti-prion capabilities for the future.

The impact of a catastrophic storm event on benthic macroinvertebrate communities in upland headwater streams and potential implications for ecological diversity and assessment of ecological status

2013-04-03T11:35:14Z

The impact of a catastrophic storm event on benthic macroinvertebrate communities in upland headwater streams and potential implications for ecological diversity and assessment of ecological status Feeley, Hugh B.; Davis, Stephen; Bruen, Michael; Blacklocke, Sean; Kelly-Quinn, Mary Upland headwater streams are dynamic systems, responding rapidly to changes in climatic conditions. This study examined the effects of a catastrophic rainfall event, that occured on 24 October 2011 on the east coast of Ireland, on the macroinvertebrate community composition and structure of four headwater streams in the river Liffey catchment located in the Wicklow Mountains. The ecological status before and after the storm were also evaluated. The water level and pH of each stream were recorded using continuous monitoring equipment, while rainfall data for the study period were sourced from a local weather station. Benthic macroinvertebrates were investigated before and after the storm event using Surber sampling. Results showed rapid and large increases in water level and significant declines in stream pH in response to intensive rainfall during the storm. The high water levels also caused major physical damage and abrasion in all four streams, that significantly altered instream habitats. The storm event induced significant losses to the richness and/or density of most taxonomic groups, with the exception of the Plecoptera. Furthermore, the overall community composition and structure changed significantly, most likely as a result of physical disturbance, given the relative persistence of acid-sensitive taxa and the relatively short period of harsh acidic conditions (<5 pH). Interestingly however, the ecological status of each of the four study sites, tested using Stream Risk Score (SSRS), Biological Monitoring Working Party (BMWP) and the Average Score Per Taxon (ASPT) indices, was unaltered by the loss in richness and densities. This was likely a result of the maintenance of plecopteran richness and the absence of organic pollution, thus highlighting the need to develop appropriate indices to assess the ecological status of streams and rivers affected by physical disturbance caused by large storm events. Ultimately, catastrophic storm events in upland headwater streams have potentially major implications for the maintenance of regional macroinvertebrate diversity within affected regions.

An in vivo root hair assay for determining rates of apoptotic-like programmed cell death in plants

2012-09-05T16:03:02Z

An in vivo root hair assay for determining rates of apoptotic-like programmed cell death in plants Kacprzyk, Joanna; O'Reilly, Conor; Gallois, Patric; McCabe, Paul F.; Hogg, Bridget V .; Molony, Elizabeth M.; Gallagher, Thomas F. In Arabidopsis thaliana we demonstrate that dying root hairs provide an easy and rapid in vivo model for the morphological identification of apoptotic-like programmed cell death in plants. The model described here is transferable between the species, can be used to investigate the rates of AL-PCD in response to various treatments and to identify modulation of AL-PCD rates in mutant/transgenic plant lines facilitating rapid screening of mutant populations in order to identify genes involved in AL-PCD regulation

The coupled δ13C-radiocarbon systematics of three late Glacial/early Holocene speleothems; insights into soil and cave processes at climatic transitions

2012-06-26T16:01:25Z

The coupled δ13C-radiocarbon systematics of three late Glacial/early Holocene speleothems; insights into soil and cave processes at climatic transitions Rudzka, Dominika; McDermott, Frank; Baldini, Lisa M.; Fleitmann, Dominik; Moreno, Ana; Stoll, Heather The coupled δ13C-radiocarbon systematics of three European stalagmites deposited during the Late Glacial and early Holocene were investigated to understand better how the carbon isotope systematics of speleothems respond to climate transitions. The emphasis is on understanding how speleothems may record climate-driven changes in the proportions of biogenic (soil carbon) and limestone bedrock derived carbon. At two of the three sites, the combined δ13C and 14C data argue against greater inputs of limestone carbon as the sole cause of the observed shift to higher d13C during the cold Younger Dryas. In these stalagmites (GAR-01 from La Garma cave, N. Spain and So-1 from Sofular cave, Turkey), the combined changes in δ13C and initial 14C activities suggest enhanced decomposition of old stored, more recalcitrant, soil carbon at the onset of the warmer early Holocene. Alternative explanations involving gradual temporal changes between open- and closed-system behaviour during the Late Glacial are difficult to reconcile with observed changes in speleothem δ13C and the growth rates. In contrast, a stalagmite from Pindal cave (N. Spain) indicates an abrupt change in carbon inputs linked to local hydrological and disequilibrium isotope fractionation effects, rather than climate change. For the first time, it is shown that while the initial 14C activities of all three stalagmites broadly follow the contemporaneous atmospheric 14C trends (the Younger Dryas atmospheric 14C anomaly can be clearly discerned), subtle changes in speleothem initial 14C activities are linked to climate-driven changes in soil carbon turnover at a climate transition.

Spatio-temporal modeling of TB in cattle herds

2012-06-26T15:51:08Z

Spatio-temporal modeling of TB in cattle herds Kelly, Gabrielle E. We examine spatial association of bovine TB in cattle herds using data from Ireland. Badgers (Meles meles), a protected species under the Wildlife Act 1976 (OAG 2012), have been implicated in the spread of the disease in cattle. Current disease control policies include reactive culling (in response to TB outbreaks) of badgers in the index and neighbouring farms. Kelly and More (2011) using generalized linear geostatistical models, established that TB clusters in cattle herds and estimated the practical spatial ranges at which this occurs. Here this work is extended by taking into account possible anisotropy. Changes in spatial association over two time periods are also examined. The results have direct implications for establishing scale and direction in reactive culling. They are also of import regarding the evaluation of vaccines for badgers and cattle.

Breeding bird populations of Irish peatlands : capsule peatlands are very important habitats for birds despite low species diversity

2012-05-22T15:02:09Z

Breeding bird populations of Irish peatlands : capsule peatlands are very important habitats for birds despite low species diversity Bracken, Fintan; McMahon, Barry J.; Whelan, John Aims to describe the variation in breeding bird populations that occur on different types of Irish peatlands and their associated habitat characteristics. Methods: Bird abundance and diversity were compared between four peatland habitat types: fens; raised bogs; Atlantic blanket bogs; and montane blanket bogs at twelve study sites using transects. Various measures of habitat quality were also taken at each location. Results: Only 21 species were recorded during the study with Meadow Pipit Anthus pratensis and Sky Lark Alauda arvensis accounting for over 80% of all birds recorded. Fens had greater bird species diversity and densities than the other three peatland types. Raised bogs, Atlantic blanket bogs and montane blanket bogs were very similar in terms of their avian diversity. Each of the recorded bird species were associated with different aspects of the peatland habitat. Conclusion: This study shows that despite the relatively low avian species diversity of Irish peatlands, they are of enormous conservation value due to the presence of species of high conservation concern such as Red Grouse (Willow Ptarmigan) Lagopus lagopus and Eurasian Curlew Numenius arquata.

Effects of set-aside management on birds breeding in lowland Ireland

2012-05-22T14:53:46Z

Effects of set-aside management on birds breeding in lowland Ireland Bracken, Fintan; Bolger, Thomas Farmland birds have suffered a severe decline in recent years throughout Europe including Ireland. Agricultural intensification is believed to be the main cause and this has led to the introduction of agri-environmental schemes, of which set-aside is a part. Bird abundance and diversity were compared between set-aside and adjacent tillage or grassland at 18 locations. The set-aside sites were also assigned to one of four management types: rotational set-aside, non-rotational set-aside, first year set-aside that was productive grassland in the previous year, and long-term set-aside that was grazed by animals in winter. Species diversity and the abundances of skylark, meadow pipit and woodpigeon were significantly greater in set-aside sites. Species diversity was not significantly different between set-aside management types and meadow pipit, skylark, pheasant, house sparrow, magpie, snipe and starling were closely associated with non-rotational set-aside, which also contained significantly larger numbers of these species compared to the other set-aside types. This study shows that set-aside does enhance bird diversity and abundance and that, in Ireland, the most effective form of set-aside is non-rotational. It also shows that the most appropriate form of set-aside will vary from situation to situation and that a one size fits all view should not be taken in the development of agri-environmental schemes.

The botanical dance of death : programmed cell death in plants

2012-01-30T12:25:41Z

The botanical dance of death : programmed cell death in plants Kacprzyk, Joanna; Daly, Cara T.; McCabe, Paul F. Programmed cell death (PCD) describes a small number of processes that result in a highly controlled, and organised, form of cellular destruction, activated in every part of the plant, throughout its entire life cycle. For example, PCD is a critical component of many vegetative and reproductive developmental processes, senescence programmes, pathogen defence mechanisms and stress responses. Cell destruction can manifest as apoptotic-like, necrotic or autophagic cell death and these processes are likely to overlap extensively, sharing several regulatory mechanisms. Several of the key PCD regulators and signals have now been revealed, for example, many cell organelles, including mitochondria, chloroplasts, Golgi apparatus, endoplasmic reticulum and vacuoles have been shown to have a role in controlling PCD activation. Following activation the actual dismantling of the cell appears to involve cell death proteases including those with caspase-like, or metacaspase, activity. This review will examine the current state of knowledge about the regulation of events during plant PCD. We will describe numerous examples of developmental or environmentally-induced deaths and outline their potential as models systems for use in PCD research programmes. Similarly, a range of techniques and in vitro model systems and that can be used to identify, and quantify, rates of plant PCD are reviewed. These model systems and techniques can be used to identify the underlying signals and events that drive and regulate PCD and ultimately reveal the steps necessary for the botanical dance of death.