Dual effects of the PI3K inhibitor ZSTK474 on multidrug efflux pumps in resistant cancer cells

ZSTK474 is a potent phosphoinositide 3-kinase (PI3K) inhibitor that reduces cell proliferation via G1-arrest. However, there is little information on the susceptibility of this anticancer drug to resistance conferred by the multidrug pumps P-glycoprotein (ABCB1) and ABCG2. We have demonstrated that ZSTK474 generated cytotoxicity in cells over-expressing either pump with potency similar to that in drug sensitive cells. In addition, the co-administration of ZSTK474 with the cytotoxic anti-cancer drugs vinblastine and mitoxantrone caused a potentiated cytotoxic effect in both drug sensitive and efflux pump expressing cells. These observations suggest that ZSTK474 is unaffected by the presence of multidrug efflux pumps and may circumvent their activities. Indeed, ZSTK474 increased the cellular accumulation of calcein-AM and mitoxantrone in cells expressing ABCB1 and ABCG2, respectively. ZSTK474 treatment also resulted in reduced expression of both efflux pumps in multidrug resistant cancer cells. Measurement of ABCB1 or ABCG2 mRNA levels demonstrated that the reduction was not due to altered transcription. Similarly, inhibitor studies showed that the proteasomal degradation pathway for ABCB1 and the lysosomal route for ABCG2 degradation were unaffected by ZSTK474. Thus the mechanism underlying reduced ABCB1 and ABCG2 levels caused by ZSTK474 was due to a reduction in overall protein synthesis; a process influenced by the PI3K pathway. In summary, ZSTK474 is not susceptible to efflux by the resistance mediators ABCB1 and ABCG2. Moreover, it inhibits the drug transport function of the pumps and leads to a reduction in their cellular expression levels. Our observations demonstrate that ZSTK474 is a powerful anticancer drug.The work in this manuscript was generously supported by a project grant (#12-0008) from Worldwide Cancer Research

UK Tourists, The Great Recession and Irish Tourism Policy. ESRI WP412. October 2011

Inbound tourism to Ireland fell sharply in 2009 and 2010. Visits to Ireland from the UK, the dominant country of origin of visitors to Ireland, did not fall faster than UK visits elsewhere. We use micro-data for UK travellers to estimate price elasticities of tourism demand for various market segments. The proposed reduction in the travel tax, and the reduction in the VAT rate for “tourism goods and services” would lead to a modest increase in visitor numbers and expenditure. However, the increase in expenditure is small compared to the foregone tax revenue

Examining Prehistoric Migration Patterns in the Palauan Archipelago: A Computer Simulated Analysis of Drift Voyaging

A number of recent genetic, linguistic, and archaeological studies have attempted to ascertain the origin of settlers to the Palauan archipelago, but it remains a complex and debated issue. To provide additional insight into colonization strategies and settlement patterns, we conducted computer simulations of drift voyages to the Palauan archipelago based on historically recorded winds and currents. Drift voyages were considered here as drifting before the wind when lost, a strategy documented for Pacific Islanders. The simulations suggest that peoples drifting before the wind from the southern Philippines would have had the most success in landfall. This finding supports the current hypothesis of human colonization to the islands of Palau. KEYWORDS: Computer simulation, drift voyaging, seafaring, colonization, Palau, Micronesia

Accumulation and distribution of doxorubicin in tumour spheroids: the influence of acidity and expression of P-glycoprotein

Purpose: The intra-tumour distribution of anticancer drugs remains an important, but often under-estimated, influence on drug efficacy. Tumour acidity and the presence of efflux pumps were examined for their influence on the distribution of doxorubicin in a solid tumour model. Methods: Anticancer drug distribution and overall accumulation was measured in tumour spheroids (TS) of varying sizes. The distribution profiles were examined in normoxic and hypoxic TS, the latter generating metabolic acidosis. Finally, the drug distribution profiles were related to efficacy using radial outgrowth assays. Results: In large tumour spheroids (TS) (d ∼500 μm), intracellular accumulation of doxorubicin was restricted to cells in the outermost layers and failed to accumulate within the viable cells in the intermediate hypoxic zone. A similar profile was obtained for another protonatable amine, 7-AAD. In contrast, the distribution of the non-ionisable drug (at physiological pH) BODIPY-Taxol was uniform throughout the TS. In order to independently model the hypoxic and normoxic zones of TS, we compared drug accumulation in small entirely normoxic TS (d ∼200 μm) with equivalent sized ones exposed to hypoxia in an anaerobic chamber. Exposure of TS to hypoxia caused a considerable reduction in the pH of the bathing medium and lower tissue accumulation of doxorubicin. Interstitial acidity reduces the proportion of doxorubicin in the non-ionised form. Conclusions: In TS, the accumulation and distribution of doxorubicin was influenced by both the expression of P-glycoprotein and hypoxia-induced acidity. Therefore, optimisation of doxorubicin chemotherapy for hypoxic tumours will require circumvention of both of these crucial pharmacokinetic determinants

Fractional boundary value problems: Analysis and numerical methods

This is the author's PDF of an article published in Fractional calculus and applied analysis 2011. The original publication is available at www.springerlink.comThis journal article discusses nonlinear boundary value problems.Fundacao para a Ciencia e Tecnologi

SEDLIN forms homodimers: characterisation of SEDLIN mutations and their interactions with transcription factors MBP1, PITX1 and SF1

BACKGROUND SEDLIN, a 140 amino acid subunit of the Transport Protein Particle (TRAPP) complex, is ubiquitously expressed and interacts with the transcription factors c-myc promoter-binding protein 1 (MBP1), pituitary homeobox 1 (PITX1) and steroidogenic factor 1 (SF1). SEDLIN mutations cause X-linked spondyloepiphyseal dysplasia tarda (SEDT). METHODOLOGY/PRINCIPAL FINDINGS We investigated the effects of 4 missense (Asp47Tyr, Ser73Leu, Phe83Ser and Val130Asp) and the most C-terminal nonsense (Gln131Stop) SEDT-associated mutations on interactions with MBP1, PITX1 and SF1 by expression in COS7 cells. Wild-type SEDLIN was present in the cytoplasm and nucleus and interacted with MBP1, PITX1 and SF1; the SEDLIN mutations did not alter these subcellular localizations or the interactions. However, SEDLIN was found to homodimerize, and the formation of dimers between wild-type and mutant SEDLIN would mask a loss in these interactions. A mammalian SEDLIN null cell-line is not available, and the interactions between SEDLIN and the transcription factors were therefore investigated in yeast, which does not endogenously express SEDLIN. This revealed that all the SEDT mutations, except Asp47Tyr, lead to a loss of interaction with MBP1, PITX1 and SF1. Three-dimensional modelling studies of SEDLIN revealed that Asp47 resides on the surface whereas all the other mutant residues lie within the hydrophobic core of the protein, and hence are likely to affect the correct folding of SEDLIN and thereby disrupt protein-protein interactions. CONCLUSIONS/SIGNIFICANCE Our studies demonstrate that SEDLIN is present in the nucleus, forms homodimers and that SEDT-associated mutations cause a loss of interaction with the transcription factors MBP1, PITX1 and SF1.This work was supported by the Oliver Bird Fund (Studentship No. RHE/00029/G), The Nuffield Foundation (J.J.), Arthritis Research Campaign (Grant ID 16438) (M.A.N. and R.V.T.), European Community Framework 7 programme grant TREAT-OA (HEALTH-F2-2008-00) (M.A.N. and R.V.T.) and the Medical Research Council (J.J., M.A.N. and R.V.T.). J.J. was an Oliver Bird funded PhD student

Recent Decisions

Comments on recent decisions by Arthur C. Callaghan, Thomas Meaney, Jr., Richard John Audino, Richard R. Murphy, Robert L. Berry, Joseph F. MacKrell, William J. Hurley, Wallace F. Neyerlin, Lawrence S. May, Jr., Richard F. Welter, Martin J. Rodgers, Anthony V. Amodio, and Robert F. McCoy

High allelic diversity in the methyltransferase gene of a phase variable type III restriction-modification system has implications for the fitness of Haemophilus influenzae

Phase variable restriction-modification (R-M) systems are widespread in Eubacteria. Haemophilus influenzae encodes a phase variable homolog of Type III R-M systems. Sequence analysis of this system in 22 non-typeable H.influenzae isolates revealed a hypervariable region in the central portion of the mod gene whereas the res gene was conserved. Maximum likelihood (ML) analysis indicated that most sites outside this hypervariable region experienced strong negative selection but evidence of positive selection for a few sites in adjacent regions. A phylogenetic analysis of 61 Type III mod genes revealed clustering of these H.influenzae mod alleles with mod genes from pathogenic Neisseriae and, based on sequence analysis, horizontal transfer of the mod–res complex between these species. Neisserial mod alleles also contained a hypervariable region and all mod alleles exhibited variability in the repeat tract. We propose that this hypervariable region encodes the target recognition domain (TRD) of the Mod protein and that variability results in alterations to the recognition sequence of this R-M system. We argue that the high allelic diversity and phase variable nature of this R-M system have arisen due to selective pressures exerted by diversity in bacteriophage populations but also have implications for other fitness attributes of these bacterial species

Estimating sampling biases in citizen science datasets

The rise of citizen science (also called community science) has led to vast quantities of species observation data collected by members of the public. Citizen science data tend to be unevenly distributed across space and time, but the treatment of sampling bias varies between studies, and interactions between different biases are often overlooked. We present a method for conceptualizing and estimating spatial and temporal sampling biases, and interactions between them. We use this method to estimate sampling biases in an example ornithological citizen science dataset from eBird in Brisbane City, Australia. We then explore the effects of these sampling biases on subsequent model inference of population trends, using both a simulation study and an application of the same trend models to the Brisbane eBird dataset. We find varying levels of sampling bias in the Brisbane eBird dataset across temporal and spatial scales, and evidence for interactions between biases. Several of the sampling biases we identified differ from those described in the literature for other datasets, with protected areas being undersampled in the city, and only limited seasonal sampling bias. We demonstrate variable performance of trend models under different sampling bias scenarios, with more complex biases being associated with typically poorer trend estimates. Sampling biases are important to consider when analysing ecological datasets, and analysts can use this method to ensure that any biologically relevant sampling biases are detected and given due consideration during analysis. With appropriate model specification, the effects of sampling biases can be reduced to yield reliable information about biodiversity.Peer reviewe