• N-methyl-N1-nitro-N-nitrosoguanidine-induced carcinogenesis: differential pattern of upper gastrointestinal tract tumours in Wistar rats after single or chronic oral doses

      Zaidi, N H; O'Connor, Peter J; Butler, W H; Cancer Research Campaign Department of Carcinogenesis, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester M20 9BX (1993)
    • N-myc gene is amplified in alveolar rhabdomyosarcomas (RMS) but not in embryonal RMS.

      Dias, P; Kumar, Patricia; Marsden, Henry B; Gattamaneni, Rao; Heighway, Jim; Kumar, Shant; Christie Hospital and Holt Radium Institute, Withington, Manchester, UK. (1990-04-15)
      DNA from 13 (6 alveolar and 7 embryonal) childhood rhabdomyosarcomas (RMS) was examined to determine the incidence and prognostic relevance of N- and c-myc genes. Southern analysis showed 5- to 20-fold amplification of N-myc gene in 4 of 6 alveolar but in none of 7 embryonal RMS (p less than 0.04; Fisher's exact test). The number of children who died with multiple- and single-copy N-myc gene was 4/4 and 5/9 respectively (p greater than 0.05; Chi-squared test). There was no statistically significant correlation between N-myc amplification and age, gender, site, stage or survival time. There was no amplification or gross rearrangement of c-myc in any of the 13 RMS.
    • N-tert-Prenylation of the indole ring improves the cytotoxicity of a short antagonist G analogue against small cell lung cancer

      Offerman, S; Kadirvel, Manikandan; Abusara, O; Bryant, J; Telfer, B; Brown, Gavin; Freeman, S; White, A; Williams, Kaye J; Aojula, H; et al. (2017)
    • NAD(P)H: quinone oxidoreductase 1 expression in kidney podocytes.

      Zappa, Francesco; Ward, Timothy H; Pedrinis, Ennio; Butler, John; McGown, Alan T; CRC Department of Drug Development, Paterson Institute for Cancer Research and Christie Hospital NHS Trust, Manchester, United Kingdom. fzappa@ticino.com (2003-03)
      NAD(P)H:quinone oxidoreductase 1 (NQO1; DT-diaphorase; DTD) is a cytosolic two-electron reductase, and compounds of the quinone family such as mitomycin C are efficiently bioactivated by this enzyme. The observation that DT-diaphorase is highly expressed in many cancerous tissues compared to normal tissues has provided us with a potentially selective target that can be exploited in the design of novel anticancer agents. Because of the relative lack of information about the cell-specific expression of DT-diaphorase, the purpose of this study was to map the distribution of this enzyme in normal human tissues. Fifteen tissue samples from normal human kidney were analyzed for expression of DT-diaphorase by immunohistochemistry (two-step indirect method). We found a specific high expression of DT-diaphorase in glomerular visceral epithelial cells (podocytes). These results suggest that a high expression of DT-diaphorase in podocytes could play a major role in the pathogenesis of renal toxicity and mitomycin C-induced hemolytic uremic syndrome, in which injury to the glomerular filtration mechanism is the primary damage, leading to a cascade of deleterious events including microangiopathic hemolytic anemia and thrombocytopenia. This observation has potential therapeutic implications because the DT-diaphorase metabolic pathway is influenced by many agents, including drugs, diet, and environmental cell factors such as pH and oxygen tension.
    • NADH autofluorescence, a new metabolic biomarker for cancer stem cells: identification of vitamin C and CAPE as natural products targeting "stemness".

      Bonuccelli, Gloria; De Francesco, Ernestina M; de Boer, Rianne; Tanowitz, Herbert B; Lisanti, Michael P; The Paterson Building, University of Manchester, Withington, United Kingdom (2017-02-16)
      Here, we assembled a broad molecular "tool-kit" to interrogate the role of metabolic heterogeneity in the propagation of cancer stem-like cells (CSCs). First, we subjected MCF7 cells to "metabolic fractionation" by flow cytometry, using fluorescent mitochondrial probes to detect PCG1α activity, as well ROS and hydrogen-peroxide (H2O2) production; NADH levels were also monitored by auto-fluorescence. Then, the various cell populations were functionally assessed for "stem cell activity", using the mammosphere assay (3D-spheroids). Our results indicate that a sub-population of MCF7 cells, with increased PGC1α activity, high mitochondrial ROS/H2O2 production and high NADH levels, all form mammospheres with a higher efficiency. Thus, it appears that mitochondrial oxidative stress and the anti-oxidant response both contribute to the promotion of mitochondrial biogenesis and oxidative metabolism in CSCs. Further validation was provided by using specific inhibitors to target metabolic processes (the NAD+ salvage pathway, glycolysis, mitochondrial protein synthesis and OXPHOS), significantly reducing CSC propagation. As a consequence, we have now identified a variety of clinically-approved drugs (stiripentol), natural products (caffeic acid phenyl ester (CAPE), ascorbic acid, silibinin) and experimental pharmaceuticals (actinonin, FK866, 2-DG), that can be used to effectively inhibit CSC activity. We discuss the use of CAPE (derived from honey-bee propolis) and Vitamin C, as potential natural therapeutic modalities. In this context, Vitamin C was ~10 times more potent than 2-DG for the targeting of CSCs. Similarly, stiripentol was between 50 to 100 times more potent than 2-DG.
    • Naloxone behaves as opioid agonist/antagonist in clonal cultures of mouse bone marrow cells.

      Krizanac-Bengez, L; Boranić, M; Testa, Nydia G; Ruder Bosković Institute, Zagreb, Croatia. (1995)
      The opioid peptide methionine (Met)-enkephalin and the opioid-receptor blocking agent naloxone were added to unseparated or to progenitor-enriched cell suspensions of mouse bone marrow before assay in clonal cultures. Bone marrow samples harvested at 18:00 hours produced more granulocyte-macrophage (GM) colonies than the 06:00 hour samples, and were more sensitive to the proliferation inhibition by both agents. Additive inhibitory effects of naloxone with the enkephalin were occasionally seen. Thus, in this experimental system, naloxone could behave as an opioid agonist. However, there were examples of naloxone diminishing (blocking) the suppressive effect of the enkephalin, as a true opioid antagonist. Significant naloxone/enkephalin interactions occurred in opioid-sensitive (18:00 h) samples of unseparated bone marrow. The interactions were virtually absent in progenitor cell-enriched populations, indicating a significant role of accessory cells in opioidergic regulation of hematopoietic progenitors.
    • Naloxone interferes with granulocytopoiesis in long-term cultures of mouse bone marrow; buffering by the stromal layer.

      Krizanac-Bengez, L; Boranić, M; Testa, Nydia G; Kardum, I; Ruder Bosković Institute, Zagreb, Croatia. (1994)
      Long-term cultures of mouse bone marrow cells were treated with naloxone, starting at the time of culture initiation or in the 2nd or 4th week of culture. Cell proliferation was suppressed and the ratio of immature and mature granulocytes to macrophages diminished by naloxone treatment. The effect depended on the timing of naloxone addition to the cultures and on its concentration, with a bell-shaped dose-response curve. High and low concentrations of naloxone (10(-4), 10(-6), 10(-14) M) interfered with hematopoiesis more strongly than the intermediate concentrations (10(-8) to 10(-12) M). Early cultures lacking the stromal layer were more sensitive to naloxone than the cultures with established stroma. The bell-shaped dose-response curve has been attributed to an interplay of specific (opioid-receptor-mediated) and nonspecific mechanisms. Opioidergic mechanisms apparently participate in the regulation of hematopoiesis.
    • Nanofluidic allele-specific digital PCR method for quantifying IDH1 and IDH2 mutation burden in acute myeloid leukemia.

      Wiseman, Daniel H; Somervaille, Tim C P; Leukemia Biology Laboratory, Cancer Research UK Manchester Institute, The University of Manchester, Manchester, M20 4BX, UK (2017)
      Precise quantitation of allelic burden for a pathogenic mutation has diverse clinical and research applications but can be difficult to achieve with conventional qPCR-based techniques, especially at lower mutant allele frequencies. Digital PCR overcomes many of the limitations of qPCR and can be highly quantitative even for single-nucleotide variants, with distinct advantages over next-generation sequencing approaches. Here we describe a method combining the principles of TaqMan(®)-chemistry SNP genotyping with microfluidic digital PCR to generate a highly sensitive, quantitative allele-specific digital PCR assay for the six most common IDH1 and IDH2 mutations encountered in myeloid malignancy. The concept and approach could easily be applied to other suitable SNVs.
    • Nanosecond flash photolysis of rhodopsin.

      Bensasson, R; Land, Edward J; Truscott, T G; E.R. 98 Laboratoire de Chimie Physique de la Faculté des Sciences, Université de Paris VI, 91405-Orsay, France (1975-12-25)
    • Naptumomab estafenatox: targeted immunotherapy with a novel immunotoxin.

      Eisen, T; Hedlund, G; Forsberg, G; Hawkins, Robert E; Cambridge University Health Partners, Addenbrooke's Hospital, Cambridge, UK, tgqe2@medschl.cam.ac.uk. (2014-02)
      Improvement of cancer therapy by introducing new concepts is still urgent even though there have been major advancements lately. Immunotherapy is well on the way to becoming an established tool in the cancer treatment armory. It seems that a combination of (1) activation of immune effector cells and selective targeting of them to tumors and (2) the inhibition of immune suppression often induced by the tumor itself are necessary to achieve the therapeutic goal. The immunotoxin naptumomab estafenatox was developed in an effort to activate and target the patient's own T cells to their tumor, by fusing a superantigen (SAg) variant that activates T lymphocytes to the Fab moiety of a tumor-reactive monoclonal antibody. Naptumomab estafenatox targets the 5T4 tumor antigen, a 72-kDa oncofetal trophoblast protein expressed on many carcinomas, including renal cell carcinoma. The therapeutic effect is associated with activation of SAg-binding T cells. The SAg-binding T lymphocytes expand, differentiate to effector cells, and infiltrate the tumor. The therapeutic efficacy is most likely related to the dual mechanism of tumor cell killing: (1) direct lysis by cytotoxic T lymphocytes of tumor cells expressing the antigen recognized by the antibody moiety of the fusion protein and (2) secretion of cytokines eliminating antigen-negative tumor cell variants. Naptumomab estafenatox has been clinically tested in a range of solid tumors with focus on renal cell carcinoma. This review looks at the clinical experience with the new immunotoxin and its potential.
    • Natural cytotoxic reactivity of human lymphocyte subpopulations.

      Potter, M R; Moore, Michael; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1979-05)
      The spontaneous cytotoxicity of human peripheral blood lymphocyte preparations from normal donors for K562 target cells was examined. Effector cells were separated into SRBC rosette forming cell (RFC) and non-rosette forming cell (non-RFC) fractions using optimal and suboptimal rosetting procedures. RFC and non-RFC fractions both had high cytotoxic activity irrespective of the rosetting procedure. Owing to the larger size of the RFC fraction, it contained a higher proportion of the total activity in the preparation. Nylon fibre column adherent and non-adherent fractions also both produced cytotoxicity. Nylon fibre non-adherent cells separated by SRBC separation gave a RFC fraction with low activity and a non-RFC fraction with high activity. Separation of nylon fibre adherent cells gave RFC and non-RFC fractions with high cytotoxic activity. Therefore cytotoxic cells did not form a discrete subpopulation and either occur in several lymphocyte subsets or show a variable capacity to form SRBC rosettes and adhere to nylon fibre.
    • Natural cytotoxicity in humans: susceptibility of freshly isolatd tumor cells to lysis.

      Vose, Brent M; Moore, Michael; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester M20 9BX, England (1980-08)
      The cytotoxic potential of blood lymphocyters from healthy donors was tested against freshly isolated lung cancer cells and the erythroleukemia K562 cell line in short-term 51Cr release assays conducted at an effector:target ratio of 50:1. Most donors exhibited significant activity against K6-562 cells. By contrast, fresh tumor cells were refractory, only 6 of 30 showing significant cytotoxicity. The low susceptibility of these tumor cells was confirmed in third-party cold inhibition assays in which they interfered minimally with killing of K562 targets under conditions in which unlabeled K562 cells efficiently blocked cytotoxicity. Cells prepared from normal lung tissue and Raji cells also failed to inhibit killing. Although in comparison to the K562 cell line freshly isolated tumor cells were resistant, their susceptibility may not be so low as to be biologically irrelevant, inasmuch as boosting of natural killing activity by interferon induced levels of cytotoxicity against both types of target cell that were unattainable by unstimulated effectors. Interferon-boosted killers were lytic for "normal" lung cells and the Raji cell line.
    • Natural cytotoxicity of human blood monocytes: Production of monocyte cytotoxic factors (MCF) during interaction with tumor cells

      Uchida, Atsushi; Yanagawa, Etsuro; Division of Immunology, Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester M20 9BX, England. (1984)
    • Natural expression of the CD19 antigen impacts the long-term engraftment but not antitumor activity of CD19-specific engineered T cells.

      Cheadle, Eleanor J; Hawkins, Robert E; Batha, Hayley; O'Neill, Allison L; Dovedi, Simon J; Gilham, David E; Cellular Therapy Group, Department of Medical Oncology, Paterson Institute for Cancer Research, The University of Manchester, Manchester, UK. echeadle@picr.man.ac.uk (2010-02-15)
      T cells gene-modified to express chimeric Ag receptors (CARs) have shown potent antitumor activity in vivo and are in clinical trials at locations worldwide. However, CAR activity has been investigated in mouse models in which Ag expression is restricted to the tumor. To explore the impact of normal tissue expression of the target Ag, we developed a mouse CD19-specific CAR to investigate antitumor efficacy against a syngeneic B cell lymphoma cell line within a background of normal CD19(+) host B cells. Mouse T cells engrafted with the amCD19CD3zeta CAR specifically lysed A20 lymphoma targets and B cells in vitro. These T cells also eradicated a 12-d established disseminated A20 lymphoma in mice preconditioned with 6 Gy total body irradiation. In the short-term (7 d after adoptive transfer), amCD19z T cells underwent Ag-dependent proliferation in vivo with a concomitant depletion in host B cell levels. However, the levels of amCD19z CAR(+) T cells decreased significantly at later time points, at which point host B cells returned, eventually reaching normal levels. In contrast, CAR(+) T cells lacking a signaling domain or specificity for mCD19 persisted over extended periods in blood and spleen. Importantly, no overt clinical signs of autotoxicity were observed in tumor-free or tumor-bearing mice treated with amCD19z T cells over an extended period of time. These observations highlight the importance of studying the activity of CAR(+) T cells in autologous models that have the normal range of tissue expression of Ag.
    • The natural history of actinic keratoses in organ transplant recipients.

      Jiyad, Z; Marquart, L; O'Rourke, P; Green, Adèle C; Cancer and Population Studies Group, QIMR Berghofer Medical Research Institute, Brisbane, Australia (2017-01)
    • The natural history of common melanocytic naevi: a systematic review of longitudinal studies in the general population.

      Plasmeijer, E; Nguyen, T; Olsen, C; Janda, M; Soyer, P; Green, Adèle C; QIMR Berghofer Medical Research Institute, Cancer and Population studies, Brisbane, QLD, Australia (2017-05-18)
    • Natural history of HLA expression during tumour development

      Garrido, F; Cabrera, T; Concha, A; Glew, Susan S; Ruiz-Cabello, F; Stern, Peter L; Dept of Analisis Clinicos e Immunologia, Hospital Virgen de las Nieves, Universidad de Granada 18014 Granada, Spain (1993)
    • Natural HPV immunity and vaccination strategies.

      Stern, Peter L; Brown, Michael D; Stacey, Simon N; Kitchener, Henry C; Hampson, Ian N; Abdel-Hady, El-Said; Moore, James V; Department of Immunology, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, M20 4BX, Manchester, UK. (2000-10)
      BACKGROUND: the task of preventing premature death in women may be delivered by vaccinating against the high-risk papillomaviruses associated with various malignancies. OBJECTIVES: we will discuss the immune mechanisms likely to be relevant to the control of an HPV infection in the cervix and assess the limited evidence for such immune recognition in the natural history of infection. CONCLUSION: the next generation of vaccination strategies should include the use of HPV 16 early (E2 and/or E6 and/or E7) and late gene targets (L1 and L2) expressed as VLPs with their clinical and immunological evaluation aimed at therapy as well as prophylaxis. Important clinical efficacy assessment may be deliverable in relatively short-term studies by targeting patients with HPV 16 associated vulval intraepithelial neoplasia.
    • Natural immunity to tumours - theoretical predictions and biological observations

      Moore, Michael; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester M20 9BX (1985)
    • Natural killer cell activity and autologous tumor killing activity in cancer patients: overlapping involvement of effector cells as determined in two-target conjugate cytotoxicity assay.

      Uchida, Atsushi; Yanagawa, Etsuro; Paterson Institute for Cancer Research, Manchester, U.K. (1984-11)
      The relationship between natural killer (NK) cell activity and autologous tumor killing activity was examined in patients with carcinomatous pleural effusions (PE) by means of a two-target conjugate cytotoxicity assay. Enrichment of large granular lymphocyte(s) (LGL) by discontinuous Percoll gradient centrifugation resulted in an augmentation of cytotoxicity against both K562 cells and tumor cells freshly isolated from PE of the same patients in a 4-hour 51Cr release cytotoxicity assay. At the single-cell level, the LGL-enriched fraction contained an increased number of effector cells that bound to autologous tumor cells and to K562 cells, as well as an increased frequency of cells cytotoxic to these target cells. In the two-target conjugate cytotoxicity assay, a single lymphocyte in the LGL population simultaneously bound to both a fluorescein-labeled K562 cell and a nonfluorescent autologous tumor cell. A significant number of lymphocytes in these mixed two-target conjugates lysed both autologous tumor cells and K562 cells after 6 hours' incubation, although overall lysis of K562 cells was higher than that of autologous tumor cells. These results indicate that a single LGL is involved in the lysis of both autologous tumor cells and K562 cells and thus provide direct evidence of involvement of subsets of NK cells in autologous tumor cell killing.