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| dUTP |
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| CHEBI:17625 |
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| A deoxyuridine phosphate having a triphosphate group at the 5'-position. |
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This entity has been manually annotated by the ChEBI Team.
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CHEBI:10533, CHEBI:42215, CHEBI:19264, CHEBI:14095
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| ZINC000008215971 |
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Molfile
XML
SDF
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more structures >>
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call loadScript javascripts\jsmol\core\package.js call loadScript javascripts\jsmol\core\core.z.js -- required by ClazzNode call loadScript javascripts\jsmol\J\awtjs2d\WebOutputChannel.js Jmol JavaScript applet jmolApplet0_object__6989129277501265__ initializing getValue debug = null getValue logLevel = null getValue allowjavascript = null AppletRegistry.checkIn(jmolApplet0_object__6989129277501265__) call loadScript javascripts\jsmol\core\corestate.z.js viewerOptions: { "name":"jmolApplet0_object","applet":true,"documentBase":"https://www.ebi.ac.uk/chebi/searchId.do?chebiId=CHEBI%3ACHEBI%3A17625","platform":"J.awtjs2d.Platform","fullName":"jmolApplet0_object__6989129277501265__","display":"jmolApplet0_canvas2d","signedApplet":"true","appletReadyCallback":"Jmol._readyCallback","statusListener":"[J.appletjs.Jmol.MyStatusListener object]","codeBase":"https://www.ebi.ac.uk/chebi/javascripts/jsmol/","syncId":"6989129277501265","bgcolor":"#000" } (C) 2012 Jmol Development Jmol Version: 13.2.7 $Date: 2013-10-01 11:35:15 -0500 (Tue, 01 Oct 2013) $ java.vendor: j2s java.version: 0.0 os.name: j2s Access: ALL memory: 0.0/0.0 processors available: 1 useCommandThread: false appletId:jmolApplet0_object (signed) starting HoverWatcher_1 getValue emulate = null defaults = "Jmol" getValue boxbgcolor = null getValue bgcolor = #000 backgroundColor = "#000" getValue ANIMFRAMECallback = null getValue APPLETREADYCallback = Jmol._readyCallback APPLETREADYCallback = "Jmol._readyCallback" getValue ATOMMOVEDCallback = null getValue CLICKCallback = null getValue ECHOCallback = null getValue ERRORCallback = null getValue EVALCallback = null getValue HOVERCallback = null getValue LOADSTRUCTCallback = null getValue MEASURECallback = null getValue MESSAGECallback = null getValue MINIMIZATIONCallback = null getValue PICKCallback = null getValue RESIZECallback = null getValue SCRIPTCallback = null getValue SYNCCallback = null getValue STRUCTUREMODIFIEDCallback = null getValue doTranslate = null language=en_US getValue popupMenu = null getValue script = null Jmol applet jmolApplet0_object__6989129277501265__ ready call loadScript javascripts\jsmol\core\corescript.z.js call loadScript javascripts\jsmol\J\script\FileLoadThread.js starting QueueThread0_2 script 1 started starting HoverWatcher_3 starting HoverWatcher_4 The Resolver thinks Mol DUT - Ideal conformer Mrv1927 07272112563D starting HoverWatcher_5 Time for openFile(DUT - Ideal conformer Mrv1927 07272112563D 43 44 0 0 0 0 999 V2000 -0.3170 0.5140 5.1660 N 0 0 0 0 0 0 0 0 0 0 0 0 -0.9720 -0.6480 4.9950 C 0 0 0 0 0 0 0 0 0 0 0 0 -2.0780 -0.9300 5.7080 N 0 0 0 0 0 0 0 0 0 0 0 0 -2.5540 -0.0420 6.6040 C 0 0 0 0 0 0 0 0 0 0 0 0 -1.8770 1.1870 6.7900 C 0 0 0 0 0 0 0 0 0 0 0 0 -0.7670 1.4420 6.0650 C 0 0 0 0 0 0 0 0 0 0 0 0 -0.5540 -1.4590 4.1910 O 0 0 0 0 0 0 0 0 0 0 0 0 -3.5580 -0.2940 7.2460 O 0 0 0 0 0 0 0 0 0 0 0 0 0.8860 0.7910 4.3790 C 0 0 1 0 0 0 0 0 0 0 0 0 1.9690 -0.2740 4.6590 C 0 0 2 0 0 0 0 0 0 0 0 0 2.6450 -0.4920 3.2870 C 0 0 1 0 0 0 0 0 0 0 0 0 1.8910 0.4640 2.3370 C 0 0 2 0 0 0 0 0 0 0 0 0 0.6060 0.6690 2.9670 O 0 0 0 0 0 0 0 0 0 0 0 0 4.0300 -0.1470 3.3500 O 0 0 0 0 0 0 0 0 0 0 0 0 1.7130 -0.1730 0.9580 C 0 0 1 0 0 0 0 0 0 0 0 0 0.9510 0.7010 0.1230 O 0 0 0 0 0 0 0 0 0 0 0 0 0.8020 -0.0280 -1.3030 P 0 0 1 0 0 5 0 0 0 0 0 0 2.1450 -0.2650 -1.8780 O 0 0 0 0 0 0 0 0 0 0 0 0 0.0460 -1.4360 -1.1090 O 0 0 0 0 0 0 0 0 0 0 0 0 -0.0490 0.9060 -2.2980 O 0 0 0 0 0 0 0 0 0 0 0 0 -0.1580 0.1180 -3.6980 P 0 0 2 0 0 5 0 0 0 0 0 0 -0.8390 -1.1770 -3.4830 O 0 0 0 0 0 0 0 0 0 0 0 0 1.3180 -0.1410 -4.2830 O 0 0 0 0 0 0 0 0 0 0 0 0 -1.0030 1.0010 -4.7460 O 0 0 0 0 0 0 0 0 0 0 0 0 -1.0710 0.1570 -6.1160 P 0 0 1 0 0 5 0 0 0 0 0 0 -1.7380 -1.1390 -5.8640 O 0 0 0 0 0 0 0 0 0 0 0 0 -1.9070 0.9850 -7.2150 O 0 0 0 0 0 0 0 0 0 0 0 0 0.4200 -0.1030 -6.6610 O 0 0 0 0 0 0 0 0 0 0 0 0 -2.5360 -1.7750 5.5750 H 0 0 0 0 0 0 0 0 0 0 0 0 -2.2430 1.9100 7.5030 H 0 0 0 0 0 0 0 0 0 0 0 0 -0.2370 2.3740 6.1940 H 0 0 0 0 0 0 0 0 0 0 0 0 1.2660 1.7870 4.6060 H 0 0 0 0 0 0 0 0 0 0 0 0 1.5130 -1.1990 5.0110 H 0 0 0 0 0 0 0 0 0 0 0 0 2.6920 0.0970 5.3860 H 0 0 0 0 0 0 0 0 0 0 0 0 2.5250 -1.5260 2.9620 H 0 0 0 0 0 0 0 0 0 0 0 0 2.4270 1.4100 2.2500 H 0 0 0 0 0 0 0 0 0 0 0 0 4.4420 -0.7560 3.9780 H 0 0 0 0 0 0 0 0 0 0 0 0 2.6900 -0.3450 0.5070 H 0 0 0 0 0 0 0 0 0 0 0 0 1.1890 -1.1230 1.0620 H 0 0 0 0 0 0 0 0 0 0 0 0 -0.8220 -1.2370 -0.7330 H 0 0 0 0 0 0 0 0 0 0 0 0 1.7260 0.7260 -4.4090 H 0 0 0 0 0 0 0 0 0 0 0 0 -1.9240 0.4460 -8.0180 H 0 0 0 0 0 0 0 0 0 0 0 0 0.8180 0.7640 -6.8120 H 0 0 0 0 0 0 0 0 0 0 0 0 1 2 1 0 0 0 0 1 6 1 0 0 0 0 1 9 1 0 0 0 0 2 3 1 0 0 0 0 2 7 2 0 0 0 0 3 4 1 0 0 0 0 3 29 1 0 0 0 0 4 5 1 0 0 0 0 4 8 2 0 0 0 0 5 6 2 0 0 0 0 5 30 1 0 0 0 0 6 31 1 0 0 0 0 9 10 1 0 0 0 0 9 13 1 0 0 0 0 9 32 1 6 0 0 0 10 11 1 0 0 0 0 10 33 1 0 0 0 0 10 34 1 0 0 0 0 11 12 1 0 0 0 0 11 14 1 0 0 0 0 11 35 1 6 0 0 0 12 13 1 0 0 0 0 12 15 1 0 0 0 0 12 36 1 6 0 0 0 14 37 1 0 0 0 0 15 16 1 0 0 0 0 15 38 1 0 0 0 0 15 39 1 0 0 0 0 17 16 1 1 0 0 0 17 18 2 0 0 0 0 17 19 1 0 0 0 0 17 20 1 0 0 0 0 19 40 1 0 0 0 0 21 20 1 1 0 0 0 21 22 2 0 0 0 0 21 23 1 0 0 0 0 21 24 1 0 0 0 0 23 41 1 0 0 0 0 24 25 1 0 0 0 0 25 26 2 0 0 0 0 25 27 1 0 0 0 0 25 28 1 0 0 0 0 27 42 1 0 0 0 0 28 43 1 0 0 0 0 M END): 16 ms reading 43 atoms ModelSet: haveSymmetry:false haveUnitcells:false haveFractionalCoord:false 1 model in this collection. Use getProperty "modelInfo" or getProperty "auxiliaryInfo" to inspect them. Default Van der Waals type for model set to Babel 43 atoms created ModelSet: not autobonding; use forceAutobond=true to force automatic bond creation Script completed Jmol script terminated
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InChI=1S/C9H15N2O14P3/c12- 5-3-8(11-2-1-7(13)10-9(11)14)23-6(5)4-22-27(18,19)25-28(20,21)24-26(15,16)17/h1-2,5-6,8,12H,3-4H2,(H,18,19)(H,20,21)(H,10,13,14)(H2,15,16,17)/t5-,6+,8+/m0/s1 |
| AHCYMLUZIRLXAA-SHYZEUOFSA-N |
| O[C@H]1C[C@@H](O[C@@H]1COP(O)(=O)OP(O)(=O)OP(O)(O)=O)N1C=CC(=O)NC1=O |
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Mus musculus
(NCBI:txid10090)
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Source: BioModels - MODEL1507180067
See:
PubMed
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Mus musculus
(NCBI:txid10090)
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From MetaboLights
See:
MetaboLights Study
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Mus musculus
(NCBI:txid10090)
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From MetaboLights
See:
MetaboLights Study
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Arabidopsis thaliana
(NCBI:txid3702)
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From MetaboLights
See:
MetaboLights Study
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Escherichia coli
(NCBI:txid562)
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See:
PubMed
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Homo sapiens
(NCBI:txid9606)
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See:
DOI
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Homo sapiens
(NCBI:txid9606)
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Found in
prostate gland
(UBERON:0002367).
See:
PubMed
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Escherichia coli metabolite
Any bacterial metabolite produced during a metabolic reaction in Escherichia coli.
human metabolite
Any mammalian metabolite produced during a metabolic reaction in humans (Homo sapiens).
mouse metabolite
Any mammalian metabolite produced during a metabolic reaction in a mouse (Mus musculus).
(via nucleoside 5'-triphoshate )
Arabidopsis thaliana metabolite
Any plant metabolite that is produced by Arabidopsis thaliana.
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View more via ChEBI Ontology
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2'-deoxyuridine 5'-(tetrahydrogen triphosphate)
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2'-deoxy-UTP
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ChEBI
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2'-deoxyuridine 5'-triphosphate
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KEGG COMPOUND
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2'-deoxyuridine triphosphate
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ChEBI
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2'-deoxyuridine-5'-triphosphorate
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HMDB
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deoxy-UTP
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HMDB
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deoxyuridine triphosphate
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ChEBI
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deoxyuridine-5'-triphosphate
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DrugBank
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dUTP
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KEGG COMPOUND
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102814-08-4
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CAS Registry Number
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ChemIDplus
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Andersson D, Svec D, Pedersen C, Henriksen JR, Ståhlberg A (2018)
Preamplification with dUTP and Cod UNG Enables Elimination of Contaminating Amplicons.
International journal of molecular sciences 19, E3185 [PubMed:30332749]
[show Abstract]
Analyzing rare DNA and RNA molecules in limited sample sizes, such as liquid biopsies and single cells, often requires preamplification, which makes downstream analyses particularly sensitive to polymerase chain reaction (PCR) generated contamination. Herein, we assessed the feasibility of performing Cod uracil-DNA N-glycosylase (Cod UNG) treatment in combination with targeted preamplification, using deoxyuridine triphosphate (dUTP) to eliminate carry-over DNA. Cod UNG can be completely and irreversibly heat inactivated, a prerequisite in preamplification methods, where any loss of amplicons is detrimental to subsequent quantification. Using 96 target assays and quantitative real-time PCR, we show that replacement of deoxythymidine triphosphate (dTTP) with dUTP in the preamplification reaction mix results in comparable dynamic range, reproducibility, and sensitivity. Moreover, Cod UNG essentially removes all uracil-containing template of most assays, regardless of initial concentration, without affecting downstream analyses. Finally, we demonstrate that the use of Cod UNG and dUTP in targeted preamplification can easily be included in the workflow for single-cell gene expression profiling. In summary, Cod UNG treatment in combination with targeted preamplification using dUTP provides a simple and efficient solution to eliminate carry-over contamination and the generation of false positives and inaccurate quantification. | Hill RLL, Vlach J, Parker LK, Christie GE, Saad JS, Dokland T (2017)
Derepression of SaPIbov1 Is Independent of φNM1 Type 2 dUTPase Activity and Is Inhibited by dUTP and dUMP.
Journal of molecular biology 429, 1570-1580 [PubMed:28400210]
[show Abstract]
Staphylococcus aureus is an opportunistic human pathogen able to transfer virulence genes to other cells through the mobilization of S. aureus pathogenicity islands (SaPIs). SaPIs are derepressed and packaged into phage-like transducing particles by helper phages like 80α or φNM1. Phages 80α and φNM1 encode structurally distinct dUTPases, Dut80α (type 1) and DutNM1 (type 2). Both dUTPases can interact with the SaPIbov1 Stl master repressor, leading to derepression and mobilization. That two structurally distinct dUTPases bind the same repressor led us to speculate that dUTPase activity may be important to the derepression process. In type 1 dUTPases, Stl binding is inhibited by dUTP. The purpose of this study was to assess the involvement of dUTP binding and dUTPase activity in derepression by DutNM1. DutNM1 activity mutants were created and tested for dUTPase activity using a novel NMR-based assay. We found that all DutNM1 null activity mutants interacted with the SaPIbov1 Stl C-terminal domain, formed DutNM1-Stl heterodimers, and caused the release of the Pstr promoter. However, promoter release was inhibited in the presence of dUTP or dUMP. We tested two φNM1 mutant phages that had null enzyme activity and found that they could still mobilize SaPIbov1. These results show that only the apo form of DutNM1 is active in Stl derepression and that dUTPase activity is not necessary for the mobilization of SaPIbov1 by DutNM1. | Bonda E, Rahav G, Kaya A, Bakhanashvili M (2016)
p53 in the mitochondria, as a trans-acting protein, provides error-correction activities during the incorporation of non-canonical dUTP into DNA.
Oncotarget 7, 73323-73336 [PubMed:27689337]
[show Abstract]
Mutations in mitochondrial DNA is an outcome of errors produced by DNA polymerase γ during replication and failure of the repair mechanism. Misincorporation of non-canonical dUTP leads to mutagenesis or apoptosis, and may contribute to the cytotoxic effects of 5'-fluorouracil chemotherapy. Tumor suppressor p53 protein in the mitochondria displays physical and functional interactions with mitochondrial DNA and polymerase γ, and by its intrinsic 3'→5' exonuclease activity can diminish the polymerization errors. Here we demonstrate the impact of p53 on incorporation of uracil into DNA examined with mitochondrial fractions, as the source of polymerase γ. p53 in mitochondria facilitates DNA damage repair functions resulting from uracil-DNA misincorporation. Our biochemical studies revealed that the procession of U:A and mismatched U:G lesions enhances in the presence of recombinant or endogenous cytoplasmic p53. p53 in mitochondria can function as an exonuclease/proofreader for polymerase γ by either decreasing the incorporation of non-canonical dUTP into DNA or by promoting the excision of incorporated nucleotide from nascent DNA, thus expanding the spectrum of DNA damage sites exploited for proofreading as a trans-acting protein. The data suggest that p53 may contribute to defense of the cells from consequences of dUTP misincorporation in both normal and tumor cells. | Roche B, Claës A, Rougeon F (2010)
Deoxyuridine triphosphate incorporation during somatic hypermutation of mouse VkOx genes after immunization with phenyloxazolone.
Journal of immunology (Baltimore, Md. : 1950) 185, 4777-4782 [PubMed:20861355]
[show Abstract]
Somatic hypermutation (SHM) of Ig genes is the result of a two-phase process initiated by activation-induced cytidine deaminase, relying on two different strategies for the introduction of mutations at CG pairs (phase I) and at AT pairs (phase II). To explain the selectivity of phase II, two mechanisms were proposed: AT-selective error-prone DNA-polymerases, deoxyuridine triphosphate (dUTP) incorporation, or both. However, there has been no experimental evidence so far of the possible involvement of the latter. We have developed a ligation-anchored PCR method based on the formation of single-strand breaks at uracils. In this study, we show the presence of uracil in hypermutating VkOx genes in wild type (AID(+/+)) mice, demonstrating that dUTP incorporation via DNA polymerases could be a major mechanism in SHM. Thus, error-prone DNA polymerases would participate in SHM via low-fidelity replication and incorporation of dUTP. | Sreekumar A, Poisson LM, Rajendiran TM, Khan AP, Cao Q, Yu J, Laxman B, Mehra R, Lonigro RJ, Li Y, Nyati MK, Ahsan A, Kalyana-Sundaram S, Han B, Cao X, Byun J, Omenn GS, Ghosh D, Pennathur S, Alexander DC, Berger A, Shuster JR, Wei JT, Varambally S, Beecher C, Chinnaiyan AM (2009)
Metabolomic profiles delineate potential role for sarcosine in prostate cancer progression.
Nature 457, 910-914 [PubMed:19212411]
[show Abstract]
Multiple, complex molecular events characterize cancer development and progression. Deciphering the molecular networks that distinguish organ-confined disease from metastatic disease may lead to the identification of critical biomarkers for cancer invasion and disease aggressiveness. Although gene and protein expression have been extensively profiled in human tumours, little is known about the global metabolomic alterations that characterize neoplastic progression. Using a combination of high-throughput liquid-and-gas-chromatography-based mass spectrometry, we profiled more than 1,126 metabolites across 262 clinical samples related to prostate cancer (42 tissues and 110 each of urine and plasma). These unbiased metabolomic profiles were able to distinguish benign prostate, clinically localized prostate cancer and metastatic disease. Sarcosine, an N-methyl derivative of the amino acid glycine, was identified as a differential metabolite that was highly increased during prostate cancer progression to metastasis and can be detected non-invasively in urine. Sarcosine levels were also increased in invasive prostate cancer cell lines relative to benign prostate epithelial cells. Knockdown of glycine-N-methyl transferase, the enzyme that generates sarcosine from glycine, attenuated prostate cancer invasion. Addition of exogenous sarcosine or knockdown of the enzyme that leads to sarcosine degradation, sarcosine dehydrogenase, induced an invasive phenotype in benign prostate epithelial cells. Androgen receptor and the ERG gene fusion product coordinately regulate components of the sarcosine pathway. Here, by profiling the metabolomic alterations of prostate cancer progression, we reveal sarcosine as a potentially important metabolic intermediary of cancer cell invasion and aggressivity. | Lamperti C, Naini AB, Lucchini V, Prelle A, Bresolin N, Moggio M, Sciacco M, Kaufmann P, DiMauro S (2005)
Muscle coenzyme Q10 level in statin-related myopathy.
Archives of neurology 62, 1709-1712 [PubMed:16286544]
[show Abstract]
BackgroundStatin drugs (3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors) reduce the level of cholesterol by inhibiting the synthesis of mevalonate, an intermediary in the cholesterol biosynthetic pathway. Use of statin drugs has been associated with a variety of skeletal muscle-related complaints. Coenzyme Q10 (CoQ10), a component of the mitochondrial respiratory chain, is also synthesized from mevalonate, and decreased muscle CoQ10 concentration may have a role in the pathogenesis of statin drug-related myopathy.ObjectivesTo measure the CoQ10 concentration and respiratory chain enzyme activities in muscle biopsy specimens from 18 patients with statin drug-related myopathy and to look for evidence of apoptosis using the TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling) assay.DesignAn open-labeled study of CoQ10 concentration in muscle from patients with increased serum creatine kinase concentrations while receiving standard statin drug therapy.SettingNeuromuscular centers at 2 academic tertiary care hospitals.ResultsMuscle structure was essentially normal in 14 patients and showed evidence of mitochondrial dysfunction and nonspecific myopathic changes in 2 patients each. Muscle CoQ10 concentration was not statistically different between patients and control subjects, but it was more than 2 SDs below the normal mean in 3 patients and more than 1 SD below normal in 7 patients. There was no TUNEL positivity in any patients.ConclusionThese data suggest that statin drug-related myopathy is associated with a mild decrease in muscle CoQ10 concentration, which does not cause histochemical or biochemical evidence of mitochondrial myopathy or morphologic evidence of apoptosis in most patients. | Lee AY, Youm YH, Kim NH, Yang H, Choi WI (2004)
Keratinocytes in the depigmented epidermis of vitiligo are more vulnerable to trauma (suction) than keratinocytes in the normally pigmented epidermis, resulting in their apoptosis.
The British journal of dermatology 151, 995-1003 [PubMed:15541077]
[show Abstract]
BackgroundVitiligo may develop following minor physical trauma. However, in autologous epidermal grafting, depigmentation of the donor (normally pigmented) site from a suction blister is rare, even in cases displaying failure of repigmentation at the recipient (depigmented) site.ObjectivesTo examine whether the suction procedure is more likely to damage keratinocytes in the depigmented than in the normally pigmented epidermis of vitiligo, and to determine what kind of damage occurs to the keratinocytes.MethodsPaired roofs of suction blisters from five patients with generalized vitiligo, five with localized and seven with segmental type, were used for the study. Multiple new lesions developed in two of the five patients with the generalized type. Apoptosis of keratinocytes in the epidermis was determined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-digoxigenin nick end labelling (TUNEL) staining, with immunohistochemistry for Bax and active caspase 3. Expression of Bcl-2, Bax, FLIP and p53, activation of caspases 3, 8 and 9, and cleavage of poly(adenosine diphosphate ribose) polymerase (PARP) in the epidermis were analysed by Western blotting in four patients with each type.ResultsApoptotic keratinocytes, which stained with TUNEL and anti-Bax and antiactive caspase 3 antibodies, were scattered in the blistered epidermis, mainly in the lower portions. The depigmented epidermis displayed significantly more apoptotic keratinocytes than the normally pigmented epidermis. The numerical difference between the paired epidermides was related to the disease activity and not to the type of lesions. The number of apoptotic keratinocytes in the normally pigmented epidermis was as high as that in the depigmented epidermis in the two patients with active generalized type vitiligo. Expression of Bax and p53 in the depigmented epidermis was higher than in the normally pigmented epidermis, whereas expression of FLIP was lower. In addition, the activation of caspases 3, 8 and 9, and cleavage of PARP, were increased in the depigmented compared with the normally pigmented epidermis. The degree of difference in expression and activation was parallel to the results of the TUNEL assay.ConclusionsThe keratinocytes in the depigmented compared with the normally pigmented epidermis of vitiligo may become apoptotic more easily after suction. | Gilhar A, Ullmann Y, Karry R, Shalaginov R, Assy B, Serafimovich S, Kalish RS (2004)
Ageing of human epidermis: the role of apoptosis, Fas and telomerase.
The British journal of dermatology 150, 56-63 [PubMed:14746617]
[show Abstract]
BackgroundAged human epidermis is characterized by morphological changes including flattening of the dermal-epidermal junction and a decrease in thickness.ObjectivesTo determine the roles of proliferation, apoptosis, Fas (CD95), Fas ligand (FasL) and telomerase in changes of human epidermis during ageing.MethodsHuman epidermis from aged subjects (n = 14; mean age 70.7 years) and young subjects (n = 14; mean age 23.4 years) was studied by histology, immunohistochemistry, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling assay for apoptotic cells and reverse transcription-polymerase chain reaction to determine epidermal thickness, proliferation (Ki-67), apoptosis, expression of Fas and FasL, and telomerase activity.ResultsAged skin was associated with thinning of the epidermis, decreased proliferation, and increased apoptosis below the granular layer. This was associated with increased epidermal expression of Fas and FasL. Telomerase activity was similar in aged and young epidermis.ConclusionsFas/FasL-mediated apoptosis, along with decreased proliferation, may have a role in changes of human epidermis during ageing. Telomerase activity did not appear to be limiting in young vs. old human epidermis. | Russell J, O'Donoghue JA, Finn R, Koziorowski J, Ruan S, Humm JL, Ling CC (2002)
Iodination of annexin V for imaging apoptosis.
Journal of nuclear medicine : official publication, Society of Nuclear Medicine 43, 671-677 [PubMed:11994533]
[show Abstract]
UnlabelledOur goal in this investigation was to develop a method for iodinating annexin V that would be suitable for the in vivo detection of apoptosis.MethodsAnnexin V was iodinated with (125)I using 2 different techniques: direct iodination with IODO-BEADS, resulting in the iodination of tyrosine residues; and use of the Bolton-Hunter reagent, which binds to lysine. The active fraction of the labeled preparation was purified by affinity chromatography. We assessed thyroid accumulation of free iodide by comparing mice with blocked and unblocked thyroids. We tested the ability of iodinated annexin V to bind apoptotic cells in vitro using irradiated neuroblastoma cells and immobilized phosphatidylserine and in vivo using C3H mice subjected to whole-body irradiation.ResultsThe efficiency of IODO-BEADS iodination was just below 30%; with the Bolton-Hunter protocol we were able to achieve 40% efficiency. When the IODO-BEADS-labeled preparation was injected into nude mice, activity accumulated rapidly in the thyroid. Two hours after injection, uptake in the thyroid region was clearly visible on a gamma-camera scan. This uptake was absent in mice that had had their thyroids blocked. We concluded that the IODO-BEADS method of labeling resulted in a protein that was rapidly deiodinated in vivo. By contrast, when annexin V was labeled using the Bolton-Hunter protocol, there was no evidence of activity accumulating in the thyroid. The Bolton-Hunter-labeled annexin V bound to apoptotic cells and immobilized phosphatidylserine in vitro. The active fraction of Bolton-Hunter-labeled annexin V was approximately 0.75. In C3H mice given 5-Gy whole-body irradiation, there was a significant induction of apoptosis in the spleen, as measured by the terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling assay, and a 4-fold increase in (125)I activity in the spleens relative to that of the control animals.ConclusionDirect iodination of annexin V on tyrosine residues is a poor technique suffering from rapid deiodination in vivo. With Bolton-Hunter chemistry, one can produce a molecule that retains its label in vivo and binds to apoptotic cells in vitro and in vivo. | Igarashi T, Brown CR, Byrum RA, Nishimura Y, Endo Y, Plishka RJ, Buckler C, Buckler-White A, Miller G, Hirsch VM, Martin MA (2002)
Rapid and irreversible CD4+ T-cell depletion induced by the highly pathogenic simian/human immunodeficiency virus SHIV(DH12R) is systemic and synchronous.
Journal of virology 76, 379-391 [PubMed:11739702]
[show Abstract]
Highly pathogenic simian/human immunodeficiency virus chimeric viruses are known to induce a rapid, irreversible depletion of CD4+ T lymphocytes in the peripheral blood of acutely infected macaque monkeys. To more fully assess the systemic effects of this primary virus infection, specimens were collected serially between days 3 and 21 postinfection from variety of lymphoid tissues (lymph nodes, thymus, and spleen) and gastrointestinal tract and examined by DNA and RNA PCR, in situ hybridization, and immunohistochemical assays. In addition, the lymphoid tissues were evaluated by fluorescence-activated cell sorting. Virus infection was initially detected by DNA PCR on day 3 postinfection in lymph node samples and peaked on day 10 in the T-lymphocyte-rich areas of this tissue. CD4+ T-cell levels remained stable through day 10 in several lymphoid tissue specimens examined but fell precipitously between days 10 and 21. In situ terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) assays revealed the accumulation of apoptotic cells during the second week of infection in both lymph nodes and thymus, which colocalized, to a large extent, to sites of both virus replication and CD4+ T-lymphocyte loss. | Oosterhuis GJ, Mulder AB, Kalsbeek-Batenburg E, Lambalk CB, Schoemaker J, Vermes I (2000)
Measuring apoptosis in human spermatozoa: a biological assay for semen quality?
Fertility and sterility 74, 245-250 [PubMed:10927039]
[show Abstract]
Objective[1] To determine whether apoptosis can be measured in ejaculated spermatozoa by flow cytometry using the Annexin V assay, which measures expression of phosphatidylserine on the outer leaflet of the cell membrane, or the TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP [deoxy-uridine triphosphate] nick end labeling) assay, which measures occurrence of DNA strand breaks and [2] to correlate the outcome with routine semen variables and the hypoosmotic swelling (HOS) test.DesignPilot study and clinical trial.SettingLarge teaching hospital and fertility center.Patient(s)Men whose semen was studied for various reasons.Main outcome measure(s)Percentage of apoptotic spermatozoa by two different assays, percentage of necrotic spermatozoa, concentration and motility of spermatozoa, and outcome of the HOS test.Result(s)Apoptosis can be measured in spermatozoa by flow cytometry using the Annexin V assay and the TUNEL assay. Twenty percent of spermatozoa were apoptotic according to both assays. A significant inverse correlation was seen between phosphatidylserine expression (Annexin V assay) and sperm concentration (r = -0.389; P<.05) and motility (r = -0.289; P<.05). A highly significant inverse correlation was seen between DNA double-strand breaks (TUNEL assay) and sperm concentration (r = -0.629; P<.0001).Conclusion(s)Flow cytometry can easily and reliably detect phosphatidylserine expression on the outer leaflet of the cell membrane and DNA strand breaks, both of which are hallmarks of apoptosis. About 20% of ejaculated spermatozoa are apoptotic, and the concentration of spermatozoa is lower in men with more apoptotic spermatozoa. | Ge YF, Huang YF, Zhang GY, Wang XH, Xu JP (1999)
Studies on apoptosis of spermatogenic cells in normal fertile men treated with supraphysiological doses of testosterone undecanoate.
Asian journal of andrology 1, 155-158 [PubMed:11250785]
[show Abstract]
AimTo study the anti-spermatogenic mechanism of supra-physiological doses of testosterone undecanoate (TU).MethodsTwenty fertile adult men received four intramuscular injections of TU at monthly intervals, 1000 mg upon admission and 500 mg for the subsequent injections. The apoptotic germ cells in the semen were studied under light microscope with terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) and Wright-Giemsa staining methods.ResultsAfter treatment, the sperm density and the number of spermatogenic cells in the semen were significantly decreased (P < 0.01), while the apoptotic ratios of spermatocytes and spermatids increased significantly (P < 0.01) as compared with the pretreatment levels. Apoptosis was found to be augmented in the whole series of castoff spermatogenic cells.ConclusionBesides its suppressive effect on spermatogenesis through a negative feed-back mechanism, TU enhances apoptosis of spermatogenic cells, which may be an additional mechanism of its anti-spermatogenic activity. | Canman CE, Lawrence TS, Shewach DS, Tang HY, Maybaum J (1993)
Resistance to fluorodeoxyuridine-induced DNA damage and cytotoxicity correlates with an elevation of deoxyuridine triphosphatase activity and failure to accumulate deoxyuridine triphosphate.
Cancer research 53, 5219-5224 [PubMed:8221659]
[show Abstract]
Deoxyuridine triphosphate (dUTP) misincorporation and uracil misrepair have long been implicated in fluoropyrimidine-induced DNA damage; however, the enzymatic activities responsible for these lesions have not been previously identified as critical determinants of overall sensitivity to the antitumor effects of these agents. The purpose of this study was to determine whether differences in uracil misincorporation/misrepair could account for the difference in sensitivity to fluorodeoxyuridine (FdUrd)-induced cytotoxicity and DNA damage in 2 human colorectal tumor cell lines having identical sensitivities to FdUrd-induced thymidylate synthase inhibition. Compared to HT29 cells, SW620 cells were resistant to both cytotoxicity and induction of DNA double-strand breaks, as assessed by pulse field gel electrophoresis. Alkaline elution experiments demonstrated that this resistance coincided with delayed induction of DNA single-strand breaks on parental DNA and, to a lesser extent, on nascent DNA. Following treatment with FdUrd for 24 h, HT29 cells accumulated 904 +/- 273 pmol deoxyuridine triphosphate (dUTP)/10(7) cells, whereas SW620 cells accumulated 20 +/- 7 pmol dUTP. Consistent with this difference in extent of dUTP accumulation was the observation that deoxyuridine triphosphatase levels in SW620 cellular extracts were 4.4-fold higher than in HT29 extracts. The ability to accumulate dUTP, intracellular deoxyuridine triphosphatase activity, and extent of DNA damage appear to be important determinants for predicting the response to FdUrd treatment in these cell lines. | Nilsson S (1979)
Deoxyuridine triphosphate in mammalian cells.
Acta chemica Scandinavica. Series B: Organic chemistry and biochemistry 33, 610 [PubMed:231363] |
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