• UKEMS collaborative genotoxicity trial. Cytogenetic tests of 4CMB, BC and 4HMB: summary and appraisal.

      Scott, David; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester M20 9BX (Great Britain) (1982)
    • Ultrastructural and morphometric alterations in bone marrow stromal tissue after 7 Gy irradiation.

      Yamazaki, K; Allen, Terence D; Department of Pathology, School of Medicine, Keio University, Tokyo, Japan. (1991)
      To evaluate the response of marrow stroma to 7 Gy irradiation, femoral bone marrow was fixed by vascular perfusion (so as to avoid the artificial destruction of sinus endothelia), and was examined using light and electron microscopy with morphometric methods. The radiation caused a marked decrease in hematopoietic cell number (NHC) within 3 days post-irradiation, followed by total recovery of hematopoiesis, which occurred gradually over 28 days. An increased number of fat cells was seen by 7 days. During the whole course of hypoplasia and recovery, the continuity of sinus wall, three-dimensional reticular mesh work in hematopoietic parenchyma, gap junctions (GJ) between stromal cells, the adventitial cell cover of sinus wall (ACC), and the stromal cell numbers of reticular cells (RC), sinus endothelia (SE), and macrophages (MP) were maintained. The cellularity of stromal components of RC, SE, and MP seemed passively increased in contrast to a reduction in numbers of NHC. A similar tendency was observed (1) between NHC and ACC and (2) between GJ and the cellularity of fat cells, which had a statistical significant correlation (p less than 0.05; t-test). The mechanism of radio resistance in bone marrow stroma and the possible functional adaptation and cellular coordination after irradiation are discussed.
    • Ultrastructural aspects of erythropoietic differentiation in long-term bone marrow culture.

      Allen, Terence D; Dexter, T Michael; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Withington, Manchester, M20 9BX, UK (1982)
      Long-term liquid cultures of mouse bone marrow produce stem cell (CFU-S) and differentiated granulocytes for many months. Addition of AMS (anaemic mouse serum) to the cultures almost entirely eliminates the granulopoietic activity and stimulates erythropoiesis, with full erythroid maturation and the production of adult haemoglobin. Ultrastructural analysis of in situ fixed material reveals the cell shape and surface morphology of the erythroid maturation series, and the generation of erythroblastic islands in vitro. Each erythroblastic island consists of one or more synchronously maturing cohorts of erythroid cells undergoing four or five divisions between proerythroblast and normoblast. Each island is centered on a macrophage, which interacts with the developing erythroid population in several ways. Expelled nuclei are phagocytosed by te macrophage, which also has large areas of closely apposed membrane with the erythroid cells, gap junctions, and possible reciprocal vesicular activity. Changes in the adherent layer (stromal cells) also occur with the transition from granulopoiesis to erythropoiesis. There is a reduction in the endothelial cell cover, and mobilisation of lipid from the granulopoietic associated apidocytes.
    • Ultrastructural changes in Osmunda regalis Prothalli induced by X-irradiation.

      Allen, Terence D; Haigh, M V; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1973-01)
    • Ultrastructural morphology of three-dimensional colonies of cells derived from a hepatocellular carcinoma.

      Allen, Terence D; Iype, P; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1979)
      Cultured hepatocellular carcinoma cells were studied during anchorage-independent growth in semi solid medium (Methocel). The regular occurrence of mitotic figures both at the surface and within the colonies precludes the possibility of such colonies being formed by re-aggregation. The estimated population doubling time in the three-dimensional (3-D) colonies is consistent with those two-dimensional of (2-D) colonies. Structures resembling bile canaliculi were observed between the closely opposed membranes from the well packed adjacent cells. Cell surface and ultrastructural features of the colonies and individual cells are presented and comparisons made with 2-D growth of normal and malignant liver cells in vitro. The formation of 3-D colonies may not only be an assay for transformed cells but also for predicting the type of tumors produced by re-innoculation of the in vitro transformed cells.
    • Ultrastructural morphometric study of efferent nerve terminals on murine bone marrow stromal cells, and the recognition of a novel anatomical unit: the "neuro-reticular complex".

      Yamazaki, K; Allen, Terence D; Department of Pathology, School of Medicine, Keio University, Tokyo, Japan. (1990-03)
      In order to extend our understanding of the role of nerve fibers in the structure and function of bone marrow stroma, we have examined nerve terminals, arterioles, and capillaries in femoral bone marrow tissues of 50 C57BL strain mice, using electron microscopy and morphometric methods. Within the adventitia of arterioles, a particular type of cell, termed periarterial adventitial (PAA) cell, is characterized by a thin veil-like cytoplasm which concentrically surrounds both nerves and arterioles. Nerve fibers containing both unmyelinated and myelinated axons are distributed mainly between the layers of PAA cells, but are found rarely on the sinus walls or within the hematopoietic parenchyma. Quantitatively, the efferent nerve terminals with many synaptic vesicles are distributed mainly beside arterial smooth muscle cells (Type I: 58.8%) or between the layers of PAA cells (Type III: 33.2%), and rarely in hematopoietic parenchyma (Type II: 5.3%) or on sinus walls (Type IV: 2.7%). In the case of Type II-IV nerve terminals, efferent (autonomic) nerves and bone marrow stromal cells which are connected by gap junctions (sinus adventitial reticular cells, intersinusoidal reticular cells, and PAA cells) appear to constitute a potential functional unit for signal conduction. We would like to propose a new term for this anatomical unit in marrow, the "neuro-reticular complex."
    • Ultrastructural site variations in mouse epidermal organization.

      Allen, Terence D; Potten, Christopher S; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1976-07)
      Mouse dorsal, ear, tail and foot epidermis are compared according to their tissue architecture and cell kinetics. Cell proliferation is expected in terms of the daily volume of keratin replaced. The stratum corneum may be organized into vertical columns of squames, which may have minimal overlap as in dorsum and ear, or maximal overlap as in tail. Individual areas are adapted to their function both in squame fine structure and rate of cell replacement. The surface keratin loss/replacement rate is at its highest in foot and tail, and lowest in ear and dorsum. Observations on hairless mouse dorsum are also included.
    • Ultrastructure of cell loss in intestinal mucosa.

      Potten, Christopher S; Allen, Terence D; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1977-08)
    • Ultrastructure of giant plastids in a radiation induced mutant of Osmunda regalis.

      Allen, Terence D; Haigh, M V; Howard, Alma; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1973-03)
    • Ultraviolet light and melanoma.

      Craig, Sarah; Earnshaw, C; Virós, Amaya; Skin Cancer and Ageing Laboratory, CRUK Manchester Institute, The University of Manchester, UK (2018-01-30)
      Melanoma is a clinically heterogeneous disease, and current strategies for treatment of the primary tumour are based on pathological criteria alone. In the recent past, several DNA-sequencing and RNA-sequencing studies of primary and advanced melanoma samples have identified unique relationships between somatic mutations, genomic aberrations, and the genetic fingerprint of ultraviolet radiation (UVR). The recurrent patterns of genomic alterations reveal different disease pathways, drug targets and mechanisms limiting drug response. Here, we examine the known associations between the molecular categories of melanoma and the multidimensional UVR damage. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
    • Ultraviolet radiation-induced DNA damage is prognostic for outcome in melanoma.

      Trucco, Lucas D; Mundra, Piyushkumar A; Hogan, Kate; Garcia-Martinez, Pablo; Viros, Amaya; Mandal, Amit Kumar; Macagno, N; Gaudy-Marqueste, C; Allan, D; Baenke, Franziska; et al. (2018)
      Erratum in Publisher Correction: Ultraviolet radiation-induced DNA damage is prognostic for outcome in melanoma. [Nat Med. 2018] Abstract The melanoma genome is dominated by ultraviolet radiation (UVR)-induced mutations. Their relevance in disease progression is unknown. Here we classify melanomas by mutation signatures and identify ten recurrently mutated UVR signature genes that predict patient survival. We validate these findings in primary human melanomas; in mice we show that this signature is imprinted by short-wavelength UVR and that four exposures to UVR are sufficient to accelerate melanomagenesis.
    • Understanding and exploiting 5T4 oncofoetal glycoprotein expression.

      Stern, Peter L; Brazzatti, J; Sawan, S; McGinn, Owen J; Institute of Cancer Sciences, University of Manchester, UK (2014-12)
      Oncofoetal antigens are present during foetal development with generally limited expression in the adult but are upregulated in cancer. These molecules can sometimes be used to diagnose or follow treatment of tumours or as a target for different immunotherapies. The 5T4 oncofoetal glycoprotein was identified by searching for shared surface molecules of human trophoblast and cancer cells with the rationale that they may function to allow survival of the foetus as a semi-allograft in the mother or a tumour in its host, potentially influencing growth, invasion or altered immune surveillance of the host. 5T4 tumour selective expression has stimulated the development of 5T4 vaccine, 5T4 antibody targeted-superantigen and 5T4 antibody-drug therapies through preclinical and into clinical studies. It is now apparent that 5T4 expression is a marker of the use (or not) of several cellular pathways relevant to tumour growth and spread. Thus 5T4 expression is mechanistically associated with the directional movement of cells through epithelial mesenchymal transition, facilitation of CXCL12/CXCR4 chemotaxis, blocking of canonical Wnt/beta-catenin while favouring non-canonical pathway signalling. These processes are highly regulated in development and in normal adult tissues but can contribute to the spread of cancer cells. Understanding the differential impact of these pathways marked by 5T4 can potentially improve existing, or aid development of novel cancer treatment strategies.
    • Understanding cancer: from the gene to the organism. Conference on genes and cancer.

      Hagan, Iain M; Sharrocks, Andrew D; School of Biological Sciences, Stopford Building, University of Manchester, Oxford Road, Manchester M13 9PT, UK. (2002-05)
    • Understanding FOXO, new views on old transcription factors.

      Zanella, Fabian; Link, Wolfgang; Carnero, Amancio; Experimental Therapeutics programme, Spanish National Cancer Research Centre, Madrid, Spain. (2010-03-01)
      FOXO proteins are evolutionarily conserved transcription factors implicated in several fundamental cellular processes, functioning as end-point for transcriptional programs involved in apoptosis, stress response and longevity. Abrogation of FOXO function is very frequent in human cancer, therefore the mechanisms of regulation of the FOXO proteins are receiving increasing attention in cancer research. The FOXO proteins integrate regulatory inputs from a variety of upstream signaling pathways, most importantly in response to growth factor and stress signalling. Recently, FOXO factors have been established as tumor suppressors, promoting the transcription of pro-apoptotic molecules like FasL and Bim when the PI3K/Akt pathway is downregulated due to nutrient or serum starvation and cellular stress. Therefore, understanding the modulation of FOXO transcription factors will allow the design of new compounds with antitumor potential.
    • Understanding the "lethal" drivers of tumor-stroma co-evolution: Emerging role(s) for hypoxia, oxidative stress and autophagy/mitophagy in the tumor micro-environment.

      Lisanti, Michael P; Martinez-Outschoorn, U E; Chiavarina, B; Pavlides, S; Whitaker-Menezes, D; Tsirigos, A; Witkiewicz, A; Lin, Z; Balliet, R; Howell, Anthony; et al. (2010-09-19)
      We have recently proposed a new model for understanding how tumors evolve. To achieve successful "Tumor-Stroma Co-Evolution", cancer cells induce oxidative stress in adjacent fibroblasts and possibly other stromal cells. Oxidative stress in the tumor stroma mimics the effects of hypoxia, under aerobic conditions, resulting in an excess production of reactive oxygen species (ROS). Excess stromal production of ROS drives the onset of an anti-oxidant defense in adjacent cancer cells, protecting them from apoptosis. Moreover, excess stromal ROS production has a "Bystander-Effect", leading to DNA damage and aneuploidy in adjacent cancer cells, both hallmarks of genomic instability. Finally, ROS-driven oxidative stress induces autophagy and mitophagy in the tumor micro-environment, leading to the stromal over-production of recycled nutrients (including energy-rich metabolites, such as ketones and L-lactate). These recycled nutrients or chemical building blocks then help drive mitochondrial biogenesis in cancer cells, thereby promoting the anabolic growth of cancer cells (via an energy imbalance). We also show that ketones and lactate help "fuel" tumor growth and cancer cell metastasis and can act as chemo-attractants for cancer cells. We have termed this new paradigm for accelerating tumor-stroma co-evolution, "The Autophagic Tumor Stroma Model of Cancer Cell Metabolism". Heterotypic signaling in cancer-associated fibroblasts activates the transcription factors HIF1alpha and NFκB, potentiating the onset of hypoxic and inflammatory response(s), which further upregulates the autophagic program in the stromal compartment. Via stromal autophagy, this hypoxic/inflammatory response may provide a new escape mechanism for cancer cells during anti-angiogenic therapy, further exacerbating tumor recurrence and metastasis.
    • Understanding the metastatic niche: is breast cancer stem cell dormancy governed by NOTCH signalling?

      Santiago-Gómez, Angélica; Spina, Elena; Spence, Katherine; Eyre, Rachel; Alférez, Denis G; Simoes, Bruno M; Clarke, Robert B; Univ Manchester, Manchester Canc Res Ctr, Manchester, Lancs, England (2018-06-30)
    • Unexpected effects on bacterial phenotype induced by expression of a tumour-amplified human sequence.

      Heighway, Jim; Santibanez-Koref, Mauro F; Department of Cancer Genetics, Paterson Institute for Cancer Research, Christie Hospital, Manchester, UK. (1989-09-12)
      An amplified human sequence was isolated from a metastatic human lung carcinoma. A fragment of this sequence situated behind the lac promoter of pUC19 appeared to affect plasmid DNA supercoiling in certain E. coli strains. Subclones were constructed to identify the smallest region of the human insert that conferred the activity and a 369 bp fragment was identified, expression of which appeared to result in abnormal plasmid supercoiling. In order to study this phenotype, various constructs were introduced into the minicell-producing strain E. coli DS410 and an unexpected effect was observed. The bacteria, after normal early growth, clumped together in late log phase, leaving virtually clear medium. Other E. coli strains were also shown to be affected to a lesser degree. The sequence producing this effect was also mapped to the 369 bp fragment and a critical region of approximately 50 bp was identified using site-directed in vitro mutagenesis and Bal31 deletion analysis. The plasmid encoded product responsible for this phenotypic alteration did not appear to be a peptide and clumping was observed when the human DNA was expressed from several bacterial promoters. It would seem likely that this sequence encodes a biologically active RNA which affects gene expression in the host cell.
    • A unified haplotype-based method for accurate and comprehensive variant calling

      Cooke, D. P.; Wedge, David C; Lunter, G.; MRC Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, UK (2021)
      Almost all haplotype-based variant callers were designed specifically for detecting common germline variation in diploid populations, and give suboptimal results in other scenarios. Here we present Octopus, a variant caller that uses a polymorphic Bayesian genotyping model capable of modeling sequencing data from a range of experimental designs within a unified haplotype-aware framework. Octopus combines sequencing reads and prior information to phase-called genotypes of arbitrary ploidy, including those with somatic mutations. We show that Octopus accurately calls germline variants in individuals, including single nucleotide variants, indels and small complex replacements such as microinversions. Using a synthetic tumor data set derived from clean sequencing data from a sample with known germline haplotypes and observed mutations in a large cohort of tumor samples, we show that Octopus is more sensitive to low-frequency somatic variation, yet calls considerably fewer false positives than other methods. Octopus also outputs realigned evidence BAM files to aid validation and interpretation.
    • United Kingdom Radiation Oncology 1 Conference (UKRO 1): accuracy and uncertainty in radiotherapy.

      Jones, B; Aird, E G; Colyer, H; Dobbs, J; Harris, R; Hoskin, P; McKenzie, A; West, Catharine M L; Oncology Centre, Hammersmith Hospital, London, UK. (2002-04)
    • Unsatisfactory quality of E. coli asparaginase biogenerics in India: Implications for clinical outcomes in acute lymphoblastic leukaemia

      Sidhu, J.; Gogoi, M. P.; Agarwal, P.; Mukherjee, T.; Saha, D.; Bose, P.; Roy, P.; Phadke, Y.; Sonawane, B.; Paul, Pritha; et al. (2021)
      Background: The biotherapeutic asparaginase is a cornerstone of therapy in acute lymphoblastic leukaemia (ALL). With limited access to the original native Escherichia coli-derived asparaginase (EcASNase), a variety of EcASNase biogenerics are used in low-middle-income countries (LMICs). The variable quality of these biogenerics potentially influences clinical outcomes. Procedure: Seven biogeneric EcASNases (P1-P7) marketed widely in India were evaluated, with P2 as an exemplar for in vivo monitoring. Therapeutic activity of P2 (10,000 IU/m2 /dose, intramuscular, every 72 hours) was monitored during induction therapy, and drug-related toxicities recorded. Molecular identity, purity and in vitro drug activity of seven biogenerics were characterised using multimodal analyses, and findings compared with reference EcASNase (R). Results: In patients (N = 62) receiving P2, subtherapeutic asparaginase activity (<100 U/L) was observed in 66% (46/70) of trough timepoints (72 hours postdose) during induction. Twelve patients (19%), 11 with high-risk ALL, developed hypersensitivity. Isoforms of EcASNase were identified in all seven biogenerics. All generic products contained impurities with batch-to-batch variability. These included high levels of protein aggregates and host cell protein contamination. In vitro assays of EcASNase activity and leukaemia cell line cytotoxicity were not discriminatory. Conclusions: Our findings confirm widespread concerns over the unsatisfactory quality and therapeutic activity of native EcASNase biogenerics marketed in LMICs. Appropriate use of these products requires monitored studies to identify clinical suitability and determine appropriate dosing and schedule. For large parts of the world, assured access to high-quality asparaginases remains an unmet therapeutic need.