• 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.
    • NADPH oxidases are required for full platelet activation in vitro and thrombosis in vivo but dispensable for plasma coagulation and hemostasisand apoptosis

      Vara, D.; Mailer, R. K.; Tarafdar, Anuradha; Wolska, N.; Heestermans, M.; Konrath, S.; Spaeth, M.; Renné, T.; Schröder, K.; Pula, G.; et al. (2020)
      Objective: Using 3KO (triple NOX [NADPH oxidase] knockout) mice (ie, NOX1-/-/NOX2-/-/NOX4-/-), we aimed to clarify the role of this family of enzymes in the regulation of platelets in vitro and hemostasis in vivo. Approach and Results: 3KO mice displayed significantly reduced platelet superoxide radical generation, which was associated with impaired platelet aggregation, adhesion, and thrombus formation in response to the key agonists collagen and thrombin. A comparison with single-gene knockouts suggested that the phenotype of 3KO platelets is the combination of the effects of the genetic deletion of NOX1 and NOX2, while NOX4 does not show any significant function in platelet regulation. 3KO platelets displayed significantly higher levels of cGMP-a negative platelet regulator that activates PKG (protein kinase G). The inhibition of PKG substantially but only partially rescued the defective phenotype of 3KO platelets, which are responsive to both collagen and thrombin in the presence of the PKG inhibitors KT5823 or Rp-8-pCPT-cGMPs, but not in the presence of the NOS (NO synthase) inhibitor L-NG-monomethyl arginine. In vivo, triple NOX deficiency protected against ferric chloride-driven carotid artery thrombosis and experimental pulmonary embolism, while hemostasis tested in a tail-tip transection assay was not affected. Procoagulatory activity of platelets (ie, phosphatidylserine surface exposure) and the coagulation cascade in platelet-free plasma were normal. Conclusions: This study indicates that inhibiting NOXs has strong antithrombotic effects partially caused by increased intracellular cGMP but spares hemostasis. NOXs are, therefore, pharmacotherapeutic targets to develop new antithrombotic drugs without bleeding side effects.
    • 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.
    • Nano-omics: nanotechnology-based multidimensional harvesting of the blood-circulating cancerome

      Gardner, Lois; Kostarelos, K.; Mallick, P.; Dive, Caroline; Hadjidemetriou, M.; Nanomedicine Lab, Faculty of Biology, Medicine and Health, The University of Manchester, Manchester, UK (2022)
      Over the past decade, the development of 'simple' blood tests that enable cancer screening, diagnosis or monitoring and facilitate the design of personalized therapies without the need for invasive tumour biopsy sampling has been a core ambition in cancer research. Data emerging from ongoing biomarker development efforts indicate that multiple markers, used individually or as part of a multimodal panel, are required to enhance the sensitivity and specificity of assays for early stage cancer detection. The discovery of cancer-associated molecular alterations that are reflected in blood at multiple dimensions (genome, epigenome, transcriptome, proteome and metabolome) and integration of the resultant multi-omics data have the potential to uncover novel biomarkers as well as to further elucidate the underlying molecular pathways. Herein, we review key advances in multi-omics liquid biopsy approaches and introduce the 'nano-omics' paradigm: the development and utilization of nanotechnology tools for the enrichment and subsequent omics analysis of the blood-circulating cancerome.
    • 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.
    • Nanomedicine: photo-activated nanostructured titanium dioxide, as a promising anticancer agent

      Lagopati, N.; Evangelou, K.; Falaras, P.; Tsilibary, E. C.; Vasileiou, P. V. S.; Havaki, S.; Angelopoulou, A.; Pavlatou, E. A.; Gorgoulis, Vassilis G; Laboratory of Histology-Embryology, Molecular Carcinogenesis Group, Faculty of Medicine, School of Health Science, National and Kapodistrian University of Athens, 75, Mikras Asias Str., Goudi, GR 11527 Athens, Greece; (2020)
      The multivariate condition of cancer disease has been approached in various ways, by the scientific community. Recent studies focus on individualized treatments, minimizing the undesirable consequences of the conventional methods, but the development of an alternative effective therapeutic scheme remains to be held. Nanomedicine could provide a solution, filling this gap, exploiting the unique properties of innovative nanostructured materials. Nanostructured titanium dioxide (TiO2) has a variety of applications of daily routine and of advanced technology. Due to its biocompatibility, it has also a great number of biomedical applications. It is now clear that photo-excited TiO2 nanoparticles, induce generation of pairs of electrons and holes which react with water and oxygen to yield reactive oxygen species (ROS) that have been proven to damage cancer cells, triggering controlled cellular processes. The aim of this review is to provide insights into the field of nanomedicine and particularly into the wide context of TiO2-NP-mediated anticancer effect, shedding light on the achievements of nanotechnology and proposing this nanostructured material as a promising anticancer photosensitizer.
    • Nanonets for multiomics blood analysis and cancer biomarker discovery

      Gardner, L.; Rothwell, Dominic G; Dive, Caroline; Kostarelos, K.; Hadjidemetriou, M.; University of Manchester, Manchester (2021)
      Despite the tremendous potential of liquid biopsies to revolutionise cancer care, there has been limited success translating blood-circulating proteomic and genomic biomarkers into the clinic. This is fundamentally due to the extremely low concentration of tumour-derived biomolecules in blood circulation, particularly at an early disease stage, which makes the discovery phase of the biomarker pipeline extremely challenging. Nanotechnology offers a promising solution, with a nanoparticle-biomolecule enrichment tool recently developed to enrich low-abundant, low molecular weight proteins in the blood of ovarian cancer patients.[1] Proteomic analysis followed by immunoassay-based validation of selected proteins demonstrated the potential of the nanoparticle-platform proposed to discover novel biomarkers with greater specificity and sensitivity than the clinically used biomarkers. In addition, we recently confirmed the presence of cell-free DNA (cfDNA) captured onto the surface lipid nanoparticles incubated ex vivo with human plasma.[2] A significantly higher abundance of cfDNA was detected in the nanoparticle-enriched plasma samples of late-stage ovarian cancer patients compared to age-matched female controls. Proteomic analysis of the same samples also revealed tumour-specific elevations in histone proteins, which are commonly found in circulation complexed with cfDNA. These findings have highlighted the opportunity for the development of a nano-proteogenomics platform able to simultaneously purify both proteins and cell-free nucleic acids from human plasma, an important step in the discovery of novel multi-omic biomarker panels. Utilising the above patented nanotechnology, we have compared proteomic and genomic profiles derived from nanoparticle-biomolecule samples of cancer patients with age- and sex-matched controls to uncover new potential blood-based biomarkers in a proof-of-principle study. In brief, ex-vivo plasma samples were incubated with lipid-based nanoparticles and purified using a two-step size-based purification protocol. The purified samples were then analysed by label-free proteomics (LC-MS/MS) and next-generation sequencing to uncover both proteomic and genomic tumour-specific signatures, including differentially abundant proteins, genomic copy number alterations and tumour-specific mutations. This work highlights the potential of our nanotechnology-based enrichment platform to enhance the discovery of cancer-specific proteogenomic biomarker panels, a vital step in developing sensitive and specific liquid biopsies for the early detection of cancer.
    • 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)