"Histories of People and Landscape. Essays on the Sheffield Region in Memory of David Hey." Industrial Archaeology Review, ahead-of-print(ahead-of-print), p. 1

Traveling in cognito, Henry Ford and his wife, Clara, spent April 1928 in England where guided by Herbert Morton, an engineer with Fords Manchester plant they traveled around looking for old steam engines to buy and ship to Dearborn USA in order to stock the giant Science Museum he was building. He was prepared to spend 10 million dollars to achieve this. This paper based on archival sources describes their search for surviving Newcomen engines and then Morton's efforts to acquire the engines, ship them to Dearborn and rebuild them there. A second paper will describe how they acquired dozens of other engines.

This paper is a personal review of the role developments in separation science over the last four decades have played in the diagnosis and understanding of purine and pyridine metabolism particularly in man. In 1967 the separation of nucleotides was used to demonstrate a new chromatography technique. This technique became known as HPLC and which continues to dominate the analysis of purines etc. The resolution and quantitation offered by even the earliest HPLC systems completely changed our understanding of matters such as nucleotide instability in cells and tissues, diagnosis of in born errors, etc. Capillary Electrophoresis also enabled high resolution as well as the quantitation of usual analytes such allantoin. Now LC-MS dominates the diagnostic field. This paper is based on the Anne Simmonds Lecture given by the author at PP17 in Gdansk in 2017.

Background HIF-1 deficiency has marked effects on tumour glycolysis and growth. We therefore investigated the consequences of HIF-1 deficiency in mice, using the well established Hepa-1 wild-type (WT) and HIF-1β-deficient (c4) model. These mechanisms could be clinically relevant, since HIF-1 is now a therapeutic target. Methods Hepa-1 WT and c4 tumours grown in vivo were analysed by 18 FDG-PET and 19 FDG Magnetic Resonance Spectroscopy for glucose uptake; by HPLC for adenine nucleotides; by immunohistochemistry for GLUTs; by immunoblotting and by DIGE followed by tandem mass spectrometry for protein expression; and by classical enzymatic methods for enzyme activity. Results HIF-1β deficient Hepa-1 c4 tumours grew significantly more slowly than WT tumours, and (as expected) showed significantly lower expression of many glycolytic enzymes. However, HIF-1β deficiency caused no significant change in the rate of glucose uptake in c4 tumours compared to WT when assessed in vivo by measuring fluoro-deoxyglucose (FDG) uptake. Immunohistochemistry demonstrated less GLUT-1 in c4 tumours, whereas GLUT-2 (liver type) was similar to WT. Factors that might upregulate glucose uptake independently of HIF-1 (phospho-Akt, c-Myc) were shown to have either lower or similar expression in c4 compared to WT tumours. However the AMP/ATP ratio was 4.5 fold higher (p < 0.01) in c4 tumours, and phosphofructokinase-1 (PFK-1) activity, measured at prevailing cellular ATP and AMP concentrations, was up to two-fold higher in homogenates of the deficient c4 cells and tumours compared to WT (p < 0.001), suggesting that allosteric PFK activation could explain their normal level of glycolysis. Phospho AMP-Kinase was also higher in the c4 tumours. Conclusions Despite their defective HIF-1 and consequent down-regulation of glycolytic enzyme expression, Hepa-1 c4 tumours maintain glucose uptake and glycolysis because the resulting low [ATP] high [AMP] allosterically activate PFK-1. This mechanism of resistance would keep glycolysis functioning and also result in activation of AMP-Kinase and growth inhibition; it may have major implications for the therapeutic activity of HIF inhibitors in vivo . Interestingly, this control mechanism does not involve transcriptional control or proteomics, but rather the classical activation and inhibition mechanisms of glycolytic enzymes.

Reduction of nitrite (NO2−) provides a major source of nitric oxide (NO) in the circulation, especially in hypoxemic conditions. Our previous studies suggest that xanthine oxidoreductase (XOR) is an important nitrite reductase in the heart and kidney. Herein, we have demonstrated that conversion of nitrite to NO by blood vessels and RBCs was enhanced in the presence of the XOR substrate xanthine (10 μmol/L) and attenuated by the XOR inhibitor allopurinol (100 μmol/L) in acidic and hypoxic conditions only. Whereas endothelial nitric oxide synthase (eNOS) inhibition had no effect on vascular nitrite reductase activity, in RBCs L-NAME, L-NMMA, andl-arginine inhibited nitrite-derived NO production by >50% (P<0.01) at pH 7.4 and 6.8 under hypoxic conditions. Western blot and immunohistochemical analysis of RBC membranes confirmed the presence of eNOS and abundant XOR on whole RBCs. Thus, XOR and eNOS are ideally situated on the membranes of RBCs and blood vessels to generate intravascular vasodilator NO from nitrite during ischemic episodes. In addition to the proposed role of deoxyhemoglobin, our findings suggest that the nitrite reductase activity within the circulation, under hypoxic conditions (at physiological pH), is mediated by eNOS; however, as acidosis develops, a substantial role for XOR becomes evident.

Reduction of nitrite (NO(2)(-)) provides a major source of nitric oxide (NO) in the circulation, especially in hypoxemic conditions. Our previous studies suggest that xanthine oxidoreductase (XOR) is an important nitrite reductase in the heart and kidney. Herein, we have demonstrated that conversion of nitrite to NO by blood vessels and RBCs was enhanced in the presence of the XOR substrate xanthine (10 micromol/L) and attenuated by the XOR inhibitor allopurinol (100 micromol/L) in acidic and hypoxic conditions only. Whereas endothelial nitric oxide synthase (eNOS) inhibition had no effect on vascular nitrite reductase activity, in RBCs L-NAME, L-NMMA, and L-arginine inhibited nitrite-derived NO production by >50% (P<0.01) at pH 7.4 and 6.8 under hypoxic conditions. Western blot and immunohistochemical analysis of RBC membranes confirmed the presence of eNOS and abundant XOR on whole RBCs. Thus, XOR and eNOS are ideally situated on the membranes of RBCs and blood vessels to generate intravascular vasodilator NO from nitrite during ischemic episodes. In addition to the proposed role of deoxyhemoglobin, our findings suggest that the nitrite reductase activity within the circulation, under hypoxic conditions (at physiological pH), is mediated by eNOS; however, as acidosis develops, a substantial role for XOR becomes evident.

The aim of this study was to establish the cause of insensitivity of T(84) human colonic epithelial cells to 5-hydroxytryptamine (5-HT). Monolayers of T(84) cells were placed in modified Ussing chambers for measurement of short-circuit current, an index of secretion. When grown in serum-supplemented media, T(84) cells gave secretory responses to acetylcholine and forskolin but not to 5-HT. When grown in AIM V serum-free media, T(84) cells responded to 5-HT. Chromatographic analysis with fluorimetric detection showed high levels of 5-HT (1.8 microM) in the serum. This contamination is probably responsible for subsequent desensitization of T(84) cells to 5-HT.

The analysis, by slab gel electrophoresis, of internucleosomal DNA cleavage or laddering, characteristic of apoptosis in many cell systems, is labour intensive, difficult to automate and best only semi-quantitative. In this report we show that CE, using dilute solutions of hydroxyethylcellulose as a replaceable sieving matrix, can be applied to the relatively rapid analysis of DNA laddering in whole digests of apoptotic rat thymocytes. Also, using the sensitivity of laser-induced fluorescence detection and the highly sensitive nucleic acid stain YO-PRO-1, the CE method reported here can use 1000–2000 fold fewer cells than needed for traditional slab gel methods.

Purines and pyrimidines are now routinely separated by HPLC. By careful selection of chromatographic conditions which match the expected changes in hydrophobicity and/or ionic nature of the substrate and products most enzymes of the purine and pyrimidine salvage pathways can be routinely and accurately determined. Ion-paired reversed-phase systems are often the most advantageous. The relevance of such assays to biomedical analysis including their role in the diagnosis of inborn errors of metabolism is stressed.

Conference Article| August 01 1990 Application of a fluorimetric method for measuring DNA strand breaks in purified DNA LEKHA L. BHUSATE; LEKHA L. BHUSATE *Department of Rheumatology, The Medical College of St Bartholomew's Hospital, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar KARL E. HERBERT; KARL E. HERBERT *Department of Rheumatology, The Medical College of St Bartholomew's Hospital, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar DAVID PERRETT DAVID PERRETT †Department of Medicine, The Medical College of St Bartholomew's Hospital, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar Biochem Soc Trans (1990) 18 (4): 676–677. https://doi.org/10.1042/bst0180676 Article history Received: November 24 1989 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn MailTo Cite Icon Cite Get Permissions Citation LEKHA L. BHUSATE, KARL E. HERBERT, DAVID PERRETT; Application of a fluorimetric method for measuring DNA strand breaks in purified DNA. Biochem Soc Trans 1 August 1990; 18 (4): 676–677. doi: https://doi.org/10.1042/bst0180676 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBiochemical Society Transactions Search Advanced Search This content is only available as a PDF. © 1990 Biochemical Society1990 Article PDF first page preview Close Modal You do not currently have access to this content.

Fibronectin, an opsonic glycoprotein has been shown to exist in fragmented forms in serum and synovial fluid. Some fragments in synovial fluid appear to be polyethylene glycol (PEG) precipitable, suggesting incorporation into immune complexes (IC). PEG precipitation, SDS-PAGE and immunoblotting were used to determine whether PEG precipitable fragments are real or artefactual. Disease specificity of fragmentation and IC incorporation of fibronectin and other proteins were also studied using these techniques. PEG precipitable fragments do not appear to be artefactual, although some fibronectin fragments are cryoprecipitable. Protein fragments showed similar distributions in whole serum and synovial fluid, disease specific differences being confined to PEG precipitates. Rheumatoid arthritis (RA) synovial fluid PEG precipitates displayed the greatest array of fragmented immunoglobulins and fibronectin. No PEG precipitates contained albumin fragments. Protein fragments in IC may impair their effective removal from RA joints. Accumulated IC could lead to tissue damage via complement activation.

Conference Article| February 01 1989 Fibronectin fragments in rheumatoid arthritis K. E. HERBERT; K. E. HERBERT 1Department of Rheumatology and Medicine, The Medical College of St Bartholomew's Hospital, West Smithfield, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar A. M. GRIFFITHS; A. M. GRIFFITHS 1Department of Rheumatology and Medicine, The Medical College of St Bartholomew's Hospital, West Smithfield, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar D. PERRETT; D. PERRETT 1Department of Rheumatology and Medicine, The Medical College of St Bartholomew's Hospital, West Smithfield, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar P. I. MAPP; P. I. MAPP 1Department of Rheumatology and Medicine, The Medical College of St Bartholomew's Hospital, West Smithfield, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar D. L. SCOTT D. L. SCOTT 1Department of Rheumatology and Medicine, The Medical College of St Bartholomew's Hospital, West Smithfield, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar Author and article information Publisher: Portland Press Ltd Received: June 16 1988 Online ISSN: 1470-8752 Print ISSN: 0300-5127 © 1989 Biochemical Society1989 Biochem Soc Trans (1989) 17 (1): 187–188. https://doi.org/10.1042/bst0170187 Article history Received: June 16 1988 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn Email Cite Icon Cite Get Permissions Citation K. E. HERBERT, A. M. GRIFFITHS, D. PERRETT, P. I. MAPP, D. L. SCOTT; Fibronectin fragments in rheumatoid arthritis. Biochem Soc Trans 1 February 1989; 17 (1): 187–188. doi: https://doi.org/10.1042/bst0170187 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBiochemical Society Transactions Search Advanced Search Keywords: RA, rheumatoid arthritis, PEG, polyethyleneglycol, PBS, phosphate-buffered saline This content is only available as a PDF. © 1989 Biochemical Society1989 Article PDF first page preview Close Modal You do not currently have access to this content.

Conference Article| June 01 1989 A critical thiol (—SH) group on the HLA-B27 molecule LACHY McLEAN; LACHY McLEAN *Bone and Joint Research Unit, London Hospital Medical College, London E1 1AD, U.K. Search for other works by this author on: This Site PubMed Google Scholar VIVIENNE WINROW; VIVIENNE WINROW *Bone and Joint Research Unit, London Hospital Medical College, London E1 1AD, U.K. Search for other works by this author on: This Site PubMed Google Scholar DAVID BLAKE; DAVID BLAKE *Bone and Joint Research Unit, London Hospital Medical College, London E1 1AD, U.K. Search for other works by this author on: This Site PubMed Google Scholar DAVID PERRETT; DAVID PERRETT †Department of Medicine, St. Bartholomew's Hospital Medical College, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar JAMES ARCHER JAMES ARCHER *Bone and Joint Research Unit, London Hospital Medical College, London E1 1AD, U.K. Search for other works by this author on: This Site PubMed Google Scholar Biochem Soc Trans (1989) 17 (3): 485–486. https://doi.org/10.1042/bst0170485 Article history Received: November 28 1988 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn MailTo Cite Icon Cite Get Permissions Citation LACHY McLEAN, VIVIENNE WINROW, DAVID BLAKE, DAVID PERRETT, JAMES ARCHER; A critical thiol (—SH) group on the HLA-B27 molecule. Biochem Soc Trans 1 June 1989; 17 (3): 485–486. doi: https://doi.org/10.1042/bst0170485 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBiochemical Society Transactions Search Advanced Search Keywords: HLA, human leucocyte antigen, PBMC, peripheral blood mononuclear cells, LCL, lymphoblastoid cells, DTNB, 5,5′-dithiobis(2-nitrobenzoic acid), NEM, N-ethylmaleimide, pCMPSA, p-chloromercuriphenylsulphonic acid This content is only available as a PDF. © 1989 Biochemical Society1989 Article PDF first page preview Close Modal You do not currently have access to this content.

Conference Article| October 01 1988 Effect of penicillamine on luminol-dependent chemiluminescence in human leucocytes RAYMOND C. FRENCH; RAYMOND C. FRENCH *Department of Biochemistry, The Medical College of St Bartholomew's Hospital, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar KARL E. HERBERT; KARL E. HERBERT †Department of Rheumatology, The Medical College of St Bartholomew's Hospital, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar DAVID L. SCOTT; DAVID L. SCOTT †Department of Rheumatology, The Medical College of St Bartholomew's Hospital, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar DAVID PERRETT DAVID PERRETT ‡Department of Medicine, The Medical College of St Bartholomew's Hospital, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar Biochem Soc Trans (1988) 16 (5): 891. https://doi.org/10.1042/bst0160891 Article history Received: April 12 1988 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn MailTo Cite Icon Cite Get Permissions Citation RAYMOND C. FRENCH, KARL E. HERBERT, DAVID L. SCOTT, DAVID PERRETT; Effect of penicillamine on luminol-dependent chemiluminescence in human leucocytes. Biochem Soc Trans 1 October 1988; 16 (5): 891. doi: https://doi.org/10.1042/bst0160891 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBiochemical Society Transactions Search Advanced Search This content is only available as a PDF. © 1988 Biochemical Society1988 Article PDF first page preview Close Modal You do not currently have access to this content.

GTP levels were low and NAD+ levels high in purine nucleoside phosphorylase (PNP) deficient erythrocytes, in addition to the raised deoxy-GTP (dGTP) levels previously noted by others. dGTP was also identified in the PNP deficient child's lymphocytes. A further novel finding was the conversion of hypoxanthine to inosine by the PNP deficient red cells, as compared to inosine monophosphate (IMP) in controls. This has been attributed to IMP formation with subsequent breakdown, and raises interesting questions regarding the controls which normally maintain erythrocyte nucleotide pools. These findings may also explain the gross purine overproduction seen in this defect; they may likewise be related to the associated immunodeficiency, anaemia, and other clinical manifestations. The results may also have important implications for the development and clinical use of PNP inhibitors.

dATP, dADP, and dAMP equalled or exceeded the depleted levels of ATP, ADP, and AMP in erythrocytes from two children with adenosine deaminase (ADA; EC 3.5.4.4) deficiency. dATP and dADP were identified in the mononuclear cells of only one child. The levels of deoxyadenosine compounds fell dramatically after enzyme replacement therapy and were no longer detectable in the urine or in mononuclear cells. Erythrocyte adenosine nucleotide levels showed a corresponding increase. Intact erythrocytes prior to treatment contained adenine, presumed to be from deoxyadenosine degraded during extraction. Adenosine at high concentrations in vitro increased both dATP and ATP levels and decreased intracellular deoxyadenosine levels. There was no significant deamination of either [8-14C]adenosine or deoxyadenosine by intact ADA-deficient erythrocytes. About 90% of adenosine was metabolized to ATP at substrate concentrations from 10-100 microM, compared to 40-60% of deoxyadenosine metabolized to dATP. These studies suggest that (i) high intracellular deoxyadenosine levels may be necessary in vivo to sustain the raised dATP levels in ADA deficiency. (ii) When ADA is inhibited or absent, deoxyadenosine is removed rapidly from the circulation by the human erythrocyte utilizing an adenosine transport system linked to both ADA and adenosine kinase (EC 2.7.1.20).

Research Article| December 01 1981 Deoxyadenosine metabolism in the erythrocytes of children with sever, combined immunodeficiency D. Perrett; D. Perrett *Medical Unit, St. Bartholomew's Hospital, London, EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar A. Sahota; A. Sahota **University of Aston, Birmingham, U.K. Search for other works by this author on: This Site PubMed Google Scholar H. A. Simmonds; H. A. Simmonds †Purine Laboratory, Guy's Tower, London Bridge St., London, SE1 9RT, U.K. Search for other works by this author on: This Site PubMed Google Scholar K. Hugh-Jones K. Hugh-Jones ††Westminster Children's Hospital, Westminster, London, SW1, U.K. Search for other works by this author on: This Site PubMed Google Scholar Biosci Rep (1981) 1 (12): 933–944. https://doi.org/10.1007/BF01114963 Article history Received: December 04 1981 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn MailTo Cite Icon Cite Get Permissions Citation D. Perrett, A. Sahota, H. A. Simmonds, K. Hugh-Jones; Deoxyadenosine metabolism in the erythrocytes of children with sever, combined immunodeficiency. Biosci Rep 1 December 1981; 1 (12): 933–944. doi: https://doi.org/10.1007/BF01114963 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBioscience Reports Search Advanced Search This content is only available as a PDF. © 1981 The Biochemical Society1981 Article PDF first page preview Close Modal You do not currently have access to this content.

Conference Article| February 01 1978 Regulation in vivo of Phosphorylase b in Skeletal Muscle of Phosphorylase Kinase-Deficient Mice ZUBAIDAH H. A. RAHIM; ZUBAIDAH H. A. RAHIM *Biochemistry Department of St. Bartholomew's Hospital Medical College, University of London, Charterhouse Square, London EC1M 6BQ, U.K. Search for other works by this author on: This Site PubMed Google Scholar DAVID PERRETT; DAVID PERRETT †Medical Unit, St. Bartholomew's Hospital, West Smithfield, London EC1A 7BE, U.K. Search for other works by this author on: This Site PubMed Google Scholar JOHN R. GRIFFITHS JOHN R. GRIFFITHS *Biochemistry Department of St. Bartholomew's Hospital Medical College, University of London, Charterhouse Square, London EC1M 6BQ, U.K. Search for other works by this author on: This Site PubMed Google Scholar Author and article information Publisher: Portland Press Ltd Online ISSN: 1470-8752 Print ISSN: 0300-5127 © 1978 Biochemical Society1978 Biochem Soc Trans (1978) 6 (1): 164–166. https://doi.org/10.1042/bst0060164 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn Email Cite Icon Cite Get Permissions Citation ZUBAIDAH H. A. RAHIM, DAVID PERRETT, JOHN R. GRIFFITHS; Regulation in vivo of Phosphorylase b in Skeletal Muscle of Phosphorylase Kinase-Deficient Mice. Biochem Soc Trans 1 February 1978; 6 (1): 164–166. doi: https://doi.org/10.1042/bst0060164 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBiochemical Society Transactions Search Advanced Search This content is only available as a PDF. © 1978 Biochemical Society1978 Article PDF first page preview Close Modal You do not currently have access to this content.