• The degradation of 5-iododeoxyuridine and 5-bromodeoxyuridine by serum from different sources and its consequences for the use of the compounds for incorporation into DNA.

      Saffhill, Roy; Hume, W J; Paterson Laboratories, Christie Hospital, Manchester M20 9BX UK (1986-03)
      Enzymes are present in sera that convert 5-iododeoxyuridine (IdU) to deoxyuridine (dU) and 5-iodouracil (IU). Although in the presence of serum 5-bromodeoxyuridine (BrdU) is not subject to extensive debromination it is converted to 5-bromouracil (BrU) at approx. 50% of the rate for IdU. These conversions are likely brought about by the enzymes thymidylate synthetase and thymidine phosphorylase. In vivo and in culture the dU enters DNA as thymidine 5'-monophosphate (dTMP) via the de novo pathway. Deoxyuridine is often found as a contaminant of [3H]IdU and [3H]BrdU. For these reasons, complications can arise in the interpretation of experimental work using these radioactive compounds. The problems may be overcome by purifying the compounds by high performance liquid chromatography (HPLC) before use together with identification of the DNA components with which the 3H is associated by chromatographic analysis.
    • Degradation of adriamycin in aqueous sodium hydroxide: formation of a ring-A oxabicyclononenone

      Abdeen, Z; Bruce, J Malcolm; Guyan, Patricia M; Land, Edward J; Mukherjee, Tulsi; Univ., Dep. Chemistry, Manchester M13 9PL, Royaume-Uni (1985)
    • Deja Vu: EGF Receptors Drive Resistance to BRAF Inhibitors.

      Girotti, Maria Romina; Marais, Richard; Molecular Oncology Group, The Paterson Institute for Cancer Research, The University of Manchester, Manchester, United Kingdom. (2013-05)
      Summary: The promise of personalized medicine is upon us, and in some cancers, targeted therapies are rapidly becoming the mainstay of treatment for selected patients based on their molecular profile. The protein kinase BRAF is a driver oncogene in both thyroid cancer and melanoma, but while drugs that target BRAF and its downstream signaling pathway are effective in melanoma, they are ineffective in thyroid cancer. In this issue of Cancer Discovery, Montero-Conde and colleagues investigate why thyroid cancer is resistant to BRAF inhibitors despite the presence of BRAF mutation. Cancer Discov; 3(5); 487-90. ©2013 AACR.
    • The delay before onset of accelerated tumour cell repopulation during radiotherapy: a direct maximum-likelihood analysis of a collection of worldwide tumour-control data.

      Roberts, Stephen A; Hendry, Jolyon H; Department of Biomathematics and Computing, Paterson Institute for Cancer Research, Christie Hospital (NHS) Trust Withington, Manchester, UK. (1993-10)
      The worldwide collection of control data for head and neck tumours presented by Withers et al. (Withers, H.R., Taylor, J.M.G. and Maciejewski, B. Acta Oncol. 27: 131-146, 1988) was reanalysed using a model which includes an explicit lag phase before the onset of tumour clonogen repopulation. A direct maximum-likelihood approach was used and the methodology extended to include the computation of profile-likelihood confidence limits. A statistically significant (p = 0.02) lag of 29 days was obtained with 95% confidence limits covering the range 17-31 days. However, the confidence interval was disconnected, and excluded the period 21-23 days. The analysis gave a time factor of 0.66 Gy/day. The mean values confirm the conclusions drawn by the original authors using a two-stage (indirect) method, and the values are similar to those calculated here for another data set comprising 496 patients (lag period = 26 (19-33) days). However, the data set itself is retrospective, and potentially subject to a number of biases. Therefore any clinical conclusions can only be tentative. A new feature of the methodology is the computation of profile-likelihood confidence limits and this will be useful in future direct analyses of clinical data of this type. The more usually computed normal approximation to the confidence limits have been shown to be inadequate in this analysis, and either profile-likelihood limits or likelihood ratio tests must be employed to determine the significance of the model parameters.
    • Delayed chromosome changes in gamma-irradiated normal and Li-Fraumeni fibroblasts.

      Boyle, John M; Spreadborough, Anne R; Greaves, Martin J; Birch, Jillian M; Varley, Jennifer; Scott, David; CRC Cancer Genetics Group, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester M20 9BX, United Kingdom. (2002-02)
      Knockout mice with only one Trp53 allele (+/- genotype) are highly susceptible to radiation-induced cancers, possibly through numerical chromosome changes. Patients with the Li-Fraumeni syndrome, having heterozygous TP53 germline mutations (+/mut genotype), are also susceptible to spontaneous and radiogenic cancers. We have investigated the susceptibility of six Li-Fraumeni syndrome +/mut and six normal fibroblast strains to induced numerical and unstable structural aberrations at six population doublings after exposure to 3 or 6 Gy gamma rays. Four of the irradiated Li-Fraumeni syndrome strains showed small increases in both aberration types, similar to those seen in the normal strains. In two irradiated Li-Fraumeni syndrome strains, there were high levels of induced structural changes, and one of these showed a modest increase in hyperploidy. We suggest that enhanced sensitivity to delayed radiation-induced chromosome changes in Li-Fraumeni syndrome cells requires other genetic alterations in addition to TP53 heterozygosity, apparently in contrast to the situation in Trp53 heterozygous null mice. If such additional alterations occur in vivo in Li-Fraumeni syndrome patients, they may predispose them to radiogenic cancers, mainly through enhanced structural rather than numerical chromosome changes. Our findings raise questions about the validity of quantitative extrapolation of cytogenetic data from Trp53-defective mice to radiogenic cancer risk in humans.
    • Delayed DNA maturation, a possible cause of the elevated sister-chromatid exchange in Bloom's syndrome.

      Ockey, Charles H; Saffhill, Roy; Paterson Laboratories, Christie Hospital, Manchester M20 9BX UK (1986-01)
      Differences in behaviour between the 5-bromodeoxyuridine (BrdU)-substituted template strands in Bloom's syndrome (BS) and normal human fibroblasts have been investigated in order to elucidate the mechanism responsible for the elevated baseline sister-chromatid exchange (SCE) frequency in BS. Alkaline sucrose gradient analysis of the normal and BrdU-substituted DNA strands showed the former to be of higher mol. wt. and of mature size while the latter were of lower molecular size, resulting from breaks introduced during the repair of the BrdU with no differences discernible between BS and normal cells. The rates of removal of BrdU were similar in BS and normal cells, which indicates that the increased SCE level in BS is not due to different rates of repair of the BrdU. The maturation of newly synthesized DNA on a normal template is delayed in BS cells compared with normal cells although it is complete at 18 h, the time it is acting as a template for DNA synthesis. In the presence of a BrdU-substituted template the maturation although further delayed is complete in normal cells by 12 h but in BS cells is not complete even by 30 h, when the newly synthesized strand, due to cell cycle delay produced by the incorporation of BrdU, becomes a template in the next round of DNA synthesis. It is suggested that a similar delay in maturation probably occurs when a new strand containing BrdU is synthesized on a normal template in BS cells. When these strands act as a template they will contain two types of breaks--those due to BrdU repair and those due to delayed maturation. The latter will be responsible for the elevated SCEs in BS cells as the DNA replication forks move through them in a manner similar to that previously reported. The possible implications of differential delays in cell proliferation in BrdU, rates of BrdU removal and extent of DNA maturation in this syndrome are discussed.
    • Delayed-type hypersensitivity response to human papillomavirus type 16 E6 protein in a mouse model.

      Chambers, M A; Stacey, Simon N; Arrand, John R; Stanley, M A; Department of Pathology, University of Cambridge, U.K. (1994-01)
      A mouse model incorporating the epitheliotropic nature of human papillomavirus (HPV) infections has been used to study an immune response to HPV type 16 (HPV-16) E6 protein in vivo. Using a transplantation technique, a novel immortal keratinocyte cell line expressing the E6 protein has been grafted onto syngeneic mice to re-form a differentiated epithelium overlying a granulation tissue bed. By this approach the presentation of viral antigens to the immune system can be modelled in a way analogous to the natural infection. Here we report a delayed-type hypersensitivity (DTH) reaction in grafted mice challenged intradermally with a recombinant vaccinia virus expressing the HPV-16 E6 protein. The specificity of the response was confirmed by the absence of a DTH reaction to challenge with virus expressing either HPV-16 E7 or L1 protein.
    • Deletion of a common region on the long arm of chromosome 6 in acute lymphoblastic leukaemia.

      Menasce, Lia P; Orphanos, Vassilis; Santibanez-Koref, Mauro F; Boyle, John M; Harrison, Christine J; Department of Histopathology, Christie Hospital, Manchester, United Kingdom. (1994-05)
      We have characterised a region of deletion on the long arm of chromosome 6 (6q) in six cases of acute lymphoblastic leukaemia, by fluorescence in situ hybridisation, using a series of YAC clones which map to 6q. Conventional cytogenetic analysis of four of these cases had been interpreted as showing terminal deletions of 6q. We demonstrated by FISH that in all cases the deletions were interstitial. D6S246 (6q16.3) was the only marker which was missing in all six cases, indicating a common region of deletion between the markers M6P1 at 6q14-15 and FYN at 6q21. Our results suggest the presence of a tumour suppressor gene within this interval.
    • Demonstration of lymphocyte nucleoli.

      Ridway, J C; Garrett, J; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1974-04)
    • Dendritic cells infected with recombinant fowlpox virus vectors are potent and long-acting stimulators of transgene-specific class I restricted T lymphocyte activity.

      Brown, Michael D; Zhang, Y; Dermime, Said; De Wynter, Erika A; Hart, Claire A; Kitchener, Henry C; Stern, Peter L; Skinner, M A; Stacey, S N; Cancer Research Campaign Laboratories, Paterson Institute of Cancer Research, Christie Hospital, Manchester, UK. (2000-10)
      The identification of dendritic cells (DC) as the major antigen-presenting cell type of the immune system, combined with the development of procedures for their ex vivo culture, has opened possibilities for tumour immunotherapy based on the transfer of recombinant tumour antigens to DC. It is anticipated that the most effective type of response would be the stimulation of specific, MHC class I restricted cytotoxic T lymphocytes capable of recognising and destroying tumour cells. In order to make this approach possible, methods must be developed for the transfer of recombinant antigen to the DC in such a way that they will initiate an MHC class I restricted response. Here, we demonstrate that murine DC infected with a recombinant fowlpox virus (rFWPV) vector stimulate a powerful, MHC class I restricted response against a recombinant antigen. A rFWPV containing the OVA gene was constructed and used to infect the DC line DC2.4. The infected DC2.4 cells were found to stimulate the T-T cell hybridoma line RF33. 70, which responds specifically to the MHC class I restricted OVA peptide SIINFEKL. The stimulatory ability of the rFWPV-infected DC2.4 cells lasted for at least 72 h after infection and was eventually limited by proliferation of uninfected cells. By comparison, DC2.4 cells pulsed with synthetic SIINFEKL peptide stimulated RF33.70 well initially, but the stimulatory ability had declined to zero by 24 h after pulsing. FWPV infection of DC2.4 up-regulated MHC and costimulatory molecule expression. rFWPV was also found to infect both immature and mature human DC derived from cord blood CD34+ progenitors and express transgenes for up to 20 days after infection. We conclude that rFWPV shows promise as a vector for antigen gene transfer to DC in tumour immunotherapy protocols.
    • Density compensated diodes for small field dosimetry: comprehensive testing and implications for design

      Georgiou, G.; Kumar, S.; Wurfel, J. U.; Underwood, Tracy; Thompson, J. M.; Hill, M. A.; Rowbottom, C. G.; Fenwick, J. D.; Department of Molecular and Clinical Cancer Medicine, Institute of Translational Medicine, University of Liverpool,Liverpool L69 3BX (2020)
      Purpose: In small megavoltage photon fields, the accuracies of an unmodified PTW 60017-type diode dosimeter and six diodes modified by adding airgaps of thickness 0.6-1.6 mm and diameter 3.6 mm have been comprehensively characterized experimentally and computationally. The optimally thick airgap for density compensation was determined, and detectors were micro-CT imaged to investigate differences between experimentally measured radiation responses and those predicted computationally. Methods: Detectors were tested onand off-axis, at 5 and 15 cm depths in 6 and 15 MV fields ≥ 0.5 × 0.5 cm2. Computational studies were carried out using the EGSnrc/BEAMnrc Monte Carlo radiation transport code. Experimentally, radiation was delivered using a Varian TrueBeam linac and doses absorbed by water were measured using Gafchromic EBT3 film and ionization chambers, and compared with diode readings. Detector response was characterized via the [Formula: see text] formalism, choosing a 4 × 4 cm2 reference field. Results: For the unmodified 60017 diode, the maximum error in small field doses obtained from diode readings uncorrected by [Formula: see text] factors was determined as 11.9% computationally at +0.25 mm off-axis and 5 cm depth in a 15 MV 0.5 × 0.5 cm2 field, and 11.7% experimentally at -0.30 mm off-axis and 5 cm depth in the same field. A detector modified to include a 1.6 mm thick airgap performed best, with maximum computationally and experimentally determined errors of 2.2% and 4.1%. The 1.6 mm airgap deepened the modified dosimeter's effective point of measurement by 0.5 mm. For some detectors significant differences existed between responses in small fields determined computationally and experimentally, micro-CT imaging indicating that these differences were due to within-tolerance variations in the thickness of an epoxy resin layer. Conclusions: The dosimetric performance of a 60017 diode detector was comprehensively improved throughout 6 and 15 MV small photon fields via density compensation. For this approach to work well with good detector-to-detector reproducibility, tolerances on dense component dimensions should be reduced to limit associated variations of response in small fields, or these components should be modified to have more water-like densities.
    • Dependence of radiation sensitivity on oxygen-tension in oedogonium .1. Extracellular and intracellular oxygen.

      Howard, Alma; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1973)
    • Depolarized fluorescence photobleaching recovery

      Dale, Robert E; Paterson Institute for Cancer Research, Christie Hospital & Holt Radium Institute, Manchester M20 9BX (1987)
    • Depression in cancer: The many biobehavioral pathways driving tumor progression.

      Bortolato, B; Hyphantis, T; Valpione, Sara; Perini, G; Maes, M; Morris, G; Kubera, M; Köhler, C; Fernandes, B; Stubbs, B; et al. (2016-11-16)
      Major Depressive Disorder (MDD) is common among cancer patients, with prevalence rates up to four-times higher than the general population. Depression confers worse outcomes, including non-adherence to treatment and increased mortality in the oncology setting. Advances in the understanding of neurobiological underpinnings of depression have revealed shared biobehavioral mechanisms may contribute to cancer progression. Moreover, psychosocial stressors in cancer promote: (1) inflammation and oxidative/nitrosative stress; (2) a decreased immunosurveillance; and (3) a dysfunctional activation of the autonomic nervous system and of the hypothalamic-pituitaryadrenal axis. Consequently, the prompt recognition of depression among patients with cancer who may benefit of treatment strategies targeting depressive symptoms, cognitive dysfunction, fatigue and sleep disturbances, is a public health priority. Moreover, behavioral strategies aiming at reducing psychological distress and depressive symptoms, including addressing unhealthy diet and life-style choices, as well as physical inactivity and sleep dysfunction, may represent important strategies not only to treat depression, but also to improve wider cancer-related outcomes. Herein, we provide a comprehensive review of the intertwined biobehavioral pathways linking depression to cancer progression. In addition, the clinical implications of these findings are critically reviewed.
    • Deregulation of Rho GTPases in cancer.

      Porter, Andrew P; Papaioannou, Alexandra; Malliri, Angeliki; Cell Signaling Group, Cancer Research UK Manchester Institute, The University of Manchester , Manchester , UK (2016-04-22)
      In vitro and in vivo studies and evidence from human tumors have long implicated Rho GTPase signaling in the formation and dissemination of a range of cancers. Recently next generation sequencing has identified direct mutations of Rho GTPases in human cancers. Moreover, the effects of ablating genes encoding Rho GTPases and their regulators in mouse models, or through pharmacological inhibition, strongly suggests that targeting Rho GTPase signaling could constitute an effective treatment. In this review we will explore the various ways in which Rho signaling can be deregulated in human cancers.
    • Derepression of the iroquois homeodomain transcription factor gene IRX3 confers differentiation block in acute leukemia.

      Somerville, Tim D D; Simeoni, Fabrizio; Chadwick, John A; Williams, Emma L; Spencer, Gary J; Boros, K; Wirth, Christopher; Tholouli, E; Byers, R; Somervaille, Tim C P; et al. (2018-01-16)
      The Iroquois homeodomain transcription factor gene IRX3 is expressed in the developing nervous system, limb buds, and heart, and transcript levels specify obesity risk in humans. We now report a functional role for IRX3 in human acute leukemia. Although transcript levels are very low in normal human bone marrow cells, high IRX3 expression is found in ∼30% of patients with acute myeloid leukemia (AML), ∼50% with T-acute lymphoblastic leukemia, and ∼20% with B-acute lymphoblastic leukemia, frequently in association with high-level HOXA gene expression. Expression of IRX3 alone was sufficient to immortalize hematopoietic stem and progenitor cells (HSPCs) in myeloid culture and induce lymphoid leukemias in vivo. IRX3 knockdown induced terminal differentiation of AML cells. Combined IRX3 and Hoxa9 expression in murine HSPCs impeded normal T-progenitor differentiation in lymphoid culture and substantially enhanced the morphologic and phenotypic differentiation block of AML in myeloid leukemia transplantation experiments through suppression of a terminal myelomonocytic program. Likewise, in cases of primary human AML, high IRX3 expression is strongly associated with reduced myelomonocytic differentiation. Thus, tissue-inappropriate derepression of IRX3 contributes significantly to the block in differentiation, which is the pathognomonic feature of human acute leukemias.
    • Deriving absolute values of alpha and beta for dose fractionation, using dose-incidence data.

      Hendry, Jolyon H; Moore, James V; Radiobiology Section, Paterson Laboratories, CHristie Hospital and Holt Radium Institute, Manchester M20 9BX (1985-09)
      A method is described for calculating absolute "operational" values of the parameters alpha and beta that characterise dose fractionation data for tissues, by using the steepness of dose-incidence curves measured around each dose per fraction investigated. The values are deduced from published mouse lethality data after irradiation of the bone marrow (alpha = 0.9 Gy-1; beta = 0.06 Gy-2), the lung (alpha = 0.14 Gy-1; beta = 0.02 Gy-2), and the oesophagus (alpha = 0.06 Gy-1; beta = 0.004 Gy-2). With bone marrow and other hierarchical renewal tissues, the operational values apply also to the inactivation of target cells which are the stem cells in the tissue. For other tissue types the interpretation of the values is unknown. The operational values are useful in characterising the steepness of dose-incidence curves for normal tissue injury after different fractionation schedules.
    • Deriving cell survival curves from the overall responses of irradiated tumours: analysis of published data for tumour spheroids.

      Moore, James V; West, Catharine M L; Hendry, Jolyon H; Department of Radiobiology, Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute, Manchester, UK. (1987-09)
      Curves of growth delay (GD) or 'cure' after graded doses of radiation have been analysed for 16 lines of human and animal tumours grown as multicellular spheroids in vitro. Dose-survival curves were derived for those cellular units from which spheroids regrow after unsuccessful irradiation (spheroid-regenerating cellular units, SRU). For 10 sets of data from 6 spheroid lines, the Do's and extrapolation numbers of the SRU derived by GD could be compared with the response of the clonogenic cells of the spheroids. For Do, a good correlation (r = 0.910) was found between the two; this was true also for Do derived from curves of spheroid 'cure' (7 sets of data from 6 spheroid lines) and clonogenic cells (r = 0.986). Using GD, the correlation of extrapolation numbers was less good (r = 0.682), the values for SRU commonly being higher than those for clonogenic cells. This may reflect features of the growth curves of spheroids after the lower range of doses of radiation. For human and animal tumour spheroids of 250 microns or less, derived Do ranged from 0.5 to 2.5 Gy. For spheroids of 350 microns or more, derived Do for animal tumour lines ranged from 3.4 to 4.2 Gy, for human lines from 1.5 to 2.1 Gy.
    • Description and basic cell kinetics of the murine pericryptal fibroblast sheath.

      Neal, J Valerie; Potten, Christopher S; Paterson Laboratories, Christie Hospital, and Holt Radium Institute, Manchester (1981-01)
      The size of the pericryptal fibroblast sheath (PCFS) population was determined by scoring serial sections. There are 38 and 124 PCFS cells per murine small intestinal and colonic crypt, respectively. The cells of the PCFS are slightly weighted towards the lower two-thirds of the crypt in their distribution. The ratio of epithelial cells to PCFS cells is approximately 6.5:1. The in vivo cell kinetics were analysed under control and stressed (fasted-refed) conditions. The control labelling index increases from 8.9% in the small intestine and 7.0% in the colon to peaks 49% and 113% respectively above these values 24 hours after 3HTdR administration. Labelling is observed at all crypt levels equally, and no evidence of vertical migration of labelled PCFS cells was found. Colonic epithelial and PCFS cells show a similar pattern of response to feeding after a fast of 72 hours with respect to time, but a different distribution of response in terms of crypt position.
    • Design, implementation and operation of a reading center platform for clinical studies.

      Clin, L; Leitritz, M; Dietter, J; Dynowski, Marek; Burgert, O; Ueffing, M; Thies, C; School of Informatics, Reutlingen University, Germany (2017)
      Clinical reading centers provide expertise for consistent, centralized analysis of medical data gathered in a distributed context. Accordingly, appropriate software solutions are required for the involved communication and data management processes. In this work, an analysis of general requirements and essential architectural and software design considerations for reading center information systems is provided. The identified patterns have been applied to the implementation of the reading center platform which is currently operated at the Center of Ophthalmology of the University Hospital of Tübingen.