Instanton counting in Class Sk\mathcal{S}_k

We compute the instanton partition functions of $\mathcal{N}=1$ SCFTs in class $\mathcal{S}_k$. We obtain this result via orbifolding Dp/D(p-4) brane systems and calculating the partition function of the supersymmetric gauge theory on the worldvolume of $K$ D(p-4) branes. Starting with D5/D1 setups probing a $\mathbb{Z}_\ell\times \mathbb{Z}_k$ orbifold singularity we obtain the $K$ instanton partition functions of 6d $(1,0)$ theories on $\mathbb{R}^4 \times T^2$ in the presence of orbifold defects on $T^2$ via computing the 2d superconformal index of the worldvolume theory on $K$ D1 branes wrapping the $T^2$. We then reduce our results to the 5d and to the 4d instanton partition functions. For $k=1$ we check that we reproduce the known elliptic, trigonometric and rational Nekrasov partition functions. Finally, we show that the instanton partition functions of $SU(N)$ quivers in class $\mathcal{S}_k$ can be obtained from the class $\mathcal{S}$ mother theory partition functions with $SU(kN)$ gauge factors via imposing the `orbifold condition' $a_{\mathcal{A}} \rightarrow a_A e^{2\pi i j/k}$ with $\mathcal{A}=jA$ and $A=1,\dots, N$, $j=1,\dots, k$ on the Coulomb moduli and the mass parameters.Comment: 43 pages, 7 figure

Prolonged expression of the γ-H2AX DNA repair biomarker correlates with excess acute and chronic toxicity from radiotherapy treatment

The normal tissue tolerance levels to fractionated radiotherapy have been appreciated by a century of careful clinical observations and radiobiological studies in animals. During clinical fractionated radiotherapy, these normal tissue tolerance levels are respected, and severe sequelae of radiotherapy are avoided in the majority of patients. Notwithstanding, a minority of patients experience unexpectedly severe normal tissue reactions. The ability to predict which patients might form this minority would be important. We have conducted a study to develop a rapid and reliable diagnostic test to predict excessive normal tissue toxicity (NTT) in radiotherapy patients. A flow cytometric immunocytochemical assay was used to measure DNA damage in peripheral blood lymphocytes (PBL) from cancer patients exposed to 2-Gy gamma radiation. DNA damage and repair was measured by induction of cellular γ-H2AX in unirradiated and exposed cells at specific time points following exposure. In 12 cancer patients that experienced severe atypical NTT following radiotherapy, there was a failure to repair DNA double-strand breaks (DSB) as measured by γ-H2AX induction and persistence. In ten cancer patients that experienced little or no NTT and in seven normal (noncancer controls), efficient repair of DNA DSB was observed in the γ-H2AX assay. We conclude that a flow cytometric assay based on γ-H2AX induction in PBL of radiotherapy patients may represent a robust, rapid and reliable biomarker to predict NTT during radiotherapy. Further research is required with a larger patient cohort to validate this important study

Countercurrent chromatography of proteins using aqueous two-phase systems

This thesis was submitted for the degree of Doctor of Philosophy and awarded by Brunel University.The biotechnology industry requires high - capacity protein manufacturing processes that retain protein functionality. Large - scale countercurrent J- type centrifuges have been developed by the Brunel Institute for Bioengineering that successfully purify small organic molecules using aqueous - organic phase systems. Aqueous two - phase systems (ATPS) have been used historically to purify bio-molecules whilst retaining their biological function. This thesis focuses on extending CCC to the separation of proteins, using a model ATPS of PEG-1000 and potassium dihydrogen phosphate to separate a mixture of lysozyme and myoglobin. Initial studies were on the behaviour of this phase system in aJ- type CCC centrifuge fitted with a multilayer column; the variable parameters examined were centrifuge rotational speed, mobile phase flow rate and direction, and type of mobile phase. A set of optimum conditions were identified that gave good retention and stability of the phases in the column. These conditions were applied to separate a mixture of the proteins lysozyme and myoglobin in the same centrifuge. However, the proteins did not elute as predicted from their equilibrium distribution ratios. It appears that the wave - like mixing and settling behaviour of the phases in the centrifuge was insufficient for the proteins to achieve equilibrium partitioning. A centrifugal partition chromatography (CPC) centrifuge was introduced to the study. This provided full protein separation, credited to the cascade phase mixing created by this design of centrifuge. The experimental parameters were used in an experiment on a pilot - scale CPC instrument. The CPC study was extended to the isolation of a target protein (phosphomannose isomerase) from a fermentation supernatant. CPC gave partial purification of the protein with retention of enzyme activity. This thesis demonstrates the importance of phase mixing in CCC, which has led to a new column design by BIB with the potential of industrial - scale protein purifications

Hypersensitivity of BRCA1 heterozygote lymphoblastoid cells to gamma radiation and PARP inhibitors

This article is made available through the Brunel Open Access Publishing Fund. Copyright @ 2013 Bourton EC, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.PARP inhibitors can be used to induce synthetic lethality in cells with bi-allelic BRCA1 and BRCA2 mutations. However the effect of PARP inhibitors in combination with radiation on cells with mono-allelic mutations of BRCA1 and BRCA2 is unknown. We have examined the cell survival response of lymphoblastoid cells derived from normal individuals and those derived from carriers of BRCA1 and BRCA2 mutations, following exposure to ionising radiation and the PARP inhibitor Olaparib. Two lymphoblastoid cell lines from normal individuals and three with mono-allelic mutations in BRCA1 and BRCA2 were exposed to increasing doses of gamma radiation either alone or in combination with 5 μM Olaparib. Cell survival was measured using the MTT assay. Exposure to increasing doses of gamma radiation caused a reduction in cell survival of all cell types. The combined exposure to gamma radiation and 5 μM Olaparib did not enhance cell kill in normal or BRCA2 heterozygote lymphoblastoid cells but significantly enhanced cell kill in cells derived from BRCA1 carriers (P = 0.02). The treatment of cancer patients carrying mutations in the BRCA1 gene with radiotherapy and the PARP inhibitor Olaparib may significantly enhance radiation induced normal tissue toxicity in these patients.Vidal Sassoon Foundation of America and “The Balls to Cancer” Charity, Coventry, U

The PARP-1 inhibitor Olaparib causes retention of γ-H2AX foci in BRCA1 heterozygote cells following exposure to gamma radiation

This article is made available through the Brunel Open Access Publishing Fund. Copyright © 2013 Emma C. Bourton et al. This is an open access article distributed under the Creative Commons Attribution Li-cense, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.A novel treatment for cancer patients with homozygous deletions of BRCA1 and BRCA2 is to use drugs that inhibit the enzyme poly(ADP-ribose) polymerase (PARP). Specific inhibition of PARP-1 can induce synthetic lethality in irradi- ated cancer cells while theoretically leaving normal tissue unaffected. We recently demonstrated in a cell survival assay that lymphoblastoid cells with mono-allelic mutations of BRCA1 were hypersensitive to gamma radiation in the pres- ence of the PARP-1 inhibitor Olaparib compared to normal cells and mono-allelic BRCA2 cells. To determine if the enhanced radiation sensitivity was due to a persistence of DNA strand breaks, we performed γ-H2AX foci analysis in cells derived from two normal individuals, three heterozygous BRCA1 and three heterozygous BRCA2 cell lines. Cells were exposed to 2 Gy gamma radiation in the presence or absence of 5 μM Olaparib. Using immunofluorescence and imaging flow cytometry, foci were measured in untreated cells and at 0.5, 3, 5 and 24 hours post-irradiation. In all lymphoblastoid cells treated with 2 Gy gamma radiation, there was a predictable induction of DNA strand breaks, with a modest but significant retention of foci over 24 hours in irradiated cells treated with Olaparib (ANOVA P < 0.05). However, in mono-allelic BRCA1 cells, there was a failure to fully repair DNA double-strand breaks (DSB) in the pres- ence of Olaparib, evidenced by a significant retention of foci at 24 hours’ post irradiation (t-Test P < 0.05). These data show that the cellular hypersensitivity of mono-allelic BRCA1 lymphoblastoid cells to gamma radiation in the presence of the Olaparib is due to the retention of DNA DSB. These data may indicate that patients with inherited mutations in the BRCA1 gene treated with radiotherapy and PARP-1 inhibitors may experience elevated radiation-associated normal tissue toxicity.Vidal Sassoon Foundation of America

Multispectral imaging flow cytometry reveals distinct frequencies of γ-H2AX foci induction in DNA double strand break repair defective human cell lines

Copyright @ 2012 International Society for Advancement of Cytometry. The article can be accessed from the links below.This article has been made available through the Brunel Open Access Publishing Fund.The measurement of γ-H2AX foci induction in cells provides a sensitive and reliable method for the quantitation of DNA damage responses in a variety of cell types. Accurate and rapid methods to conduct such observations are desirable. In this study we have employed the novel technique of multispectral imaging flow cytometry to compare the induction and repair of γ-H2AX foci in three human cell types with different capacities for the repair of DNA double strand breaks (DSB). A repair normal fibroblast cell line MRC5-SV1, a DSB repair defective ataxia telangiectasia (AT5BIVA) cell line, and a DNA-PKcs deficient cell line XP14BRneo17 were exposed to 2 Gy gamma radiation from a 60Cobalt source. Thirty minutes following exposure we observed a dramatic induction of foci in the nuclei of these cells. After 24 hrs there was a predictable reduction on the number of foci in the MRC5-SV1 cells, consistent with the repair of DNA DSB. In the AT5BIVA cells, persistence of the foci over a 24 hour period was due to the failure in the repair of DNA DSB. However, in the DNA-PKcs defective cells (XP14BRneo17) we observed an intermediate retention of foci in the nuclei indicative of partial repair of DNA DSB. In summary, the application of imaging flow cytometry has permitted an evaluation of foci in a large number of cells (20,000) for each cell line at each time point. This provides a novel method to determine differences in repair kinetics between different cell types. We propose that imaging flow cytometry provides an alternative platform for accurate automated high through-put analysis of foci induction in a variety of cell types.This article is made available through the Brunel Open Access Publishing Fund

A critical and comparative analysis of the prevention of tax evasion through the application of law and enforcement policies in the United Kingdom and United States of America

Following the global financial crisis, and a number of tax evasion scandals, there has been an increased public demand for countries to tackle tax evasion. In response, the past two decades have seen the diffusion of international cooperation in tax matters of a nature and on a scale that was previously unthinkable, facilitating the near eradication of offshore tax evasion. Additionally, throughout this time, both the UK and US have paid greater attention to this financial crime, amending relevant laws and enforcement policies, as well as empowering and supporting tax authorities to enhance investigations. In the UK, there has not only been a stronger public appetite to combat tax evasion, but also, a desire to combat tax evasion using the procedures and penalties of the criminal justice system. In light of these contemporary national and international developments, this thesis provides a unique contribution to knowledge by comparing and evaluating the laws and enforcement policies pertaining to tax evasion in the UK and US. This thesis argues that the laws and enforcement policy pertaining to tax evasion in the UK are neither internally, nor externally, effective in combatting tax evasion and a comprehensive reform of the UK’s approach is long overdue. Significant insights can be gained from the US in improving both the internal and external effectiveness of the UK legal framework