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

Biocidal silver and silver/titania composite films grown by chemical vapour deposition

This paper describes the growth and testing of highly active biocidal films based on photocatalytically active films of TiO2, grownby thermal CVD, functionally and structurallymodified by deposition of nanostructured silver via a novel flame assisted combination CVD process. The resulting composite films are shown to be highly durable, highly photocatalytically active and are also shown to possess strong antibacterial behaviour. The deposition control, arising from the described approach, offers the potential to control the film nanostructure, which is proposed to be crucial in determining the photo and bioactivity of the combined film structure, and the transparency of the composite films. Furthermore, we show that the resultant films are active to a range of organisms, including Gram-negative and Gram-positive bacteria, and viruses. The very high-biocidal activity is above that expected from the concentrations of silver present, and this is discussed in terms of nanostructure of the titania/silver surface. These properties are especially significant when combined with the well-known durability of CVD deposited thin films, offering new opportunities for enhanced application in areas where biocidal surface functionality is sought

Photocatalytic antimicrobial activity of thin surface films of TiO2, CuO and TiO2 /CuO dual layers on Escherichia coli and bacteriophage T4

TiO2 coated surfaces are increasingly studied for their ability to inactivate microorganisms. The activity of glass coated with thin films of TiO2, CuO and hybrid CuO/TiO2 prepared by atmospheric Chemical Vapour Deposition (Ap-CVD) and TiO2 prepared by a sol-gel process was investigated using the inactivation of bacteriophage T4 as a model for inactivation of viruses. The chemical oxidising activity was also determined by measuring stearic acid oxidation. The results showed that the rate of inactivation of bacteriophage T4 increased with increasing chemical oxidising activity with the maximum rate obtained on highly active sol-gel preparations. However these were delicate and easily damaged unlike the Ap-CVD coatings. Inactivation rates were highest on CuO and CuO/TiO2 which had the lowest chemical oxidising activities. The inactivation of T4 was higher than that of Escherichia coli on low activity surfaces. The combination of photocatalysis and toxicity of copper acted synergistically to inactivate bacteriophage T4 and retained some selfcleaning activity. The presence of phosphate ions slowed inactivation but NaCl had no effect. The results show that TiO2/CuO coated surfaces are highly antiviral and may have applications in the food and healthcare industries

The human myometrium differentially expresses mTOR signalling components before and during pregnancy: Evidence for regulation by progesterone

Emerging studies implicate the signalling of the mammalian target of rapamycin (mTOR) in a number of reproductive functions. To this date, there are no data regarding the expression of mTOR signalling components in the human myometrium during pregnancy. We hypothesized that mTOR-related genes might be differentially expressed in term or preterm labour as well as in labour or non-labour myometria during pregnancy. Using quantitative RT-PCR we demonstrate for first time that there is a significant downregulation of mTOR, DEPTOR, and Raptor in preterm labouring myometria when compared to non-pregnant tissues taken from the same area (lower segment). We used an immortalized myometrial cell line (ULTR) as an in vitro model to dissect further mTOR signalling. In ULTR cells DEPTOR and Rictor had a cytoplasmic distribution, whereas mTOR and Raptor were detected in the cytoplasm and the nucleus, indicative of mTORC1 shuttling. Treatment with inflammatory cytokines caused only minor changes in gene expression of these components, whereas progesterone caused significant down-regulation. We performed a non-biased gene expression analysis of ULTR cells using Nimblegen human gene expression microarray (n = 3), and selected genes were validated by quantitative RT-PCR in progesterone treated myometrial cells. Progesterone significantly down-regulated key components of the mTOR pathway. We conclude that the human myometrium differentially expresses mTOR signalling components and they can be regulated by progesterone. This article is part of a Special Issue entitled 'Pregnancy and Steroids'.This research was funded by National Institutes of Health Grant ESO12961

The consequences of replicating in the wrong orientation: Bacterial chromosome duplication without an active replication origin

Chromosome replication is regulated in all organisms at the assembly stage of the replication machinery at specific origins. In Escherichia coli the DnaA initiator protein regulates the assembly of replication forks at oriC. This regulation can be undermined by defects in nucleic acid meta¬bolism. In cells lacking RNase HI replication initiates indepen¬dently of DnaA and oriC, presumably at persisting R-loops. A similar mechanism was assumed for origin-independent synthesis in cells lacking RecG. However, recently we suggested that this synthesis initiates at intermediates resulting from replication fork fusions. Here we present data suggesting that in cells lacking RecG or RNase HI origin-independent synthesis arises by different mechanisms, indicative of these two proteins having different roles in vivo. Our data support the idea that RNase HI processes R-loops, while RecG is required to process replication fork fusion intermediates. However, regardless of how origin-independent synthesis is initiated, a fraction of forks will proceed in an orientation opposite to normal. We show that the resulting head-on encounters with transcription threaten cell viability, especially if taking place in highly-transcribed areas. Thus, despite their different functions, RecG and RNase HI are both important factors for maintaining replication control and orientation. Their absence causes severe replication problems, highlighting the advantages of the normal chromosome arrangement, which exploits a single origin to control the number of forks and their orientation relative to transcription, and a defined termination area to contain fork fusions. Any changes to this arrangement endanger cell cycle control, chromosome dynamics and, ultimately, cell viability.This work was supported by the Royal Society (RG110414 to C.J.R.) and The Biotechnology and Biological Sciences Research Council (BB/K015729/1 to C.J.R.)

Solid state NMR and X-ray diffraction studies of α-d-galacturonic acid monohydrate

Crystalline a-d-galacturonic acid monohydrate has been studied by 13C CPMAS NMR and X-ray crystallography. The molecular dynamics were investigated by evaluating 13C spin-lattice relaxation in the rotating frame (T1?) and chemical-shift-anisotropy properties of each carbon. Only limited molecular motions can be detected in the low frequency

Dual functionality anti-reflection and biocidal coatings

A thin film combination of anti-reflection (AR) and biocidal properties would be of particular interest to reduce the transfer of infection and improve readability of public high use touch screens. In this paper we describe the development of a dual functionality film of silica (AR) and copper oxide (biocidal). Deposition was via flame assisted chemical vapour deposition (FACVD) which has the advantages of being a cost efficient atmospheric pressure technique enabling use of non-volatile precursors and that no closed reaction cell is required so making it ideal for integration into industrial production lines. The resulting films were characterized by a range of techniques including optical spectroscopy, electron microscope and X-ray fluorescence. Biocidal behavior was tested by determining the kill rate of Escherichia coli. A 3 layer stack on glass of silica/copper oxide/silica had better adhesion and lower reflection than a comparable 2 layer stack. This multilayer film led to a > 2% drop in reflection from that of uncoated glass, similar to that of silica only film. In addition, showed a > 6 log10 kill between 6 and 24 h for as deposited and annealed samples

Curriculum Infusion Through Case Studies: Engaging Undergraduate Students In Course Subject Material and Influencing Behavior Change

This study investigated infusing health promotion topics into an engineering course via problem-based case studies and lecture to assess student learning and self-reported behavior. Junior-level systems engineering students in two sections participated: one section with 52 students and one with 36. One section received a celebratory drinking case; one received distracted driving case and a lecture about hazardous drinking. Student ability ratings related to the course subject matter generally improved with both cases. The lecture appeared to enhance health promotion knowledge. Students self-reported behavior change with both cases. Case studies as a form of curriculum infusion for health promotion topics show promise. The use of case studies overall was well-received by students and coupled with lecture material can increase student health promotion knowledge and behavior change

Interphase chromosome positioning in in vitro porcine cells and ex vivo porcine tissues

Copyright @ 2012 The Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and 85 reproduction in any medium, provided the original author and source are credited. The article was made available through the Brunel University Open Access Publishing Fund.BACKGROUND: In interphase nuclei of a wide range of species chromosomes are organised into their own specific locations termed territories. These chromosome territories are non-randomly positioned in nuclei which is believed to be related to a spatial aspect of regulatory control over gene expression. In this study we have adopted the pig as a model in which to study interphase chromosome positioning and follows on from other studies from our group of using pig cells and tissues to study interphase genome re-positioning during differentiation. The pig is an important model organism both economically and as a closely related species to study human disease models. This is why great efforts have been made to accomplish the full genome sequence in the last decade. RESULTS: This study has positioned most of the porcine chromosomes in in vitro cultured adult and embryonic fibroblasts, early passage stromal derived mesenchymal stem cells and lymphocytes. The study is further expanded to position four chromosomes in ex vivo tissue derived from pig kidney, lung and brain. CONCLUSIONS: It was concluded that porcine chromosomes are also non-randomly positioned within interphase nuclei with few major differences in chromosome position in interphase nuclei between different cell and tissue types. There were also no differences between preferred nuclear location of chromosomes in in vitro cultured cells as compared to cells in tissue sections. Using a number of analyses to ascertain by what criteria porcine chromosomes were positioned in interphase nuclei; we found a correlation with DNA content.This study is partly supported by Sygen International PLC