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Journal of Medicinal Chemistry Sep 1967
Topics: Deoxyuridine; Oxides; Thymidine
PubMed: 6048518
DOI: 10.1021/jm00317a059 -
Proceedings of the National Academy of... Feb 2008We have developed a method to detect DNA synthesis in proliferating cells, based on the incorporation of 5-ethynyl-2'-deoxyuridine (EdU) and its subsequent detection by...
We have developed a method to detect DNA synthesis in proliferating cells, based on the incorporation of 5-ethynyl-2'-deoxyuridine (EdU) and its subsequent detection by a fluorescent azide through a Cu(I)-catalyzed [3 + 2] cycloaddition reaction ("click" chemistry). Detection of the EdU label is highly sensitive and can be accomplished in minutes. The small size of the fluorescent azides used for detection results in a high degree of specimen penetration, allowing the staining of whole-mount preparations of large tissue and organ explants. In contrast to BrdU, the method does not require sample fixation or DNA denaturation and permits good structural preservation. We demonstrate the use of the method in cultured cells and in the intestine and brain of whole animals.
Topics: Animals; DNA; DNA Replication; Deoxyuridine; Mice; Molecular Structure; NIH 3T3 Cells; Reproducibility of Results; Sensitivity and Specificity; Time Factors
PubMed: 18272492
DOI: 10.1073/pnas.0712168105 -
Methods in Molecular Biology (Clifton,... 2021The principles and practice of a methodology of cell cycle analysis that allows the estimation of the absolute length (in units of time) of all cell cycle stages (G1, S,...
The principles and practice of a methodology of cell cycle analysis that allows the estimation of the absolute length (in units of time) of all cell cycle stages (G1, S, and G2) are detailed herein. This methodology utilizes flow cytometry to take full advantage of the excellent stoichiometric properties of click chemistry. This allows detection, via azide-fluorochrome coupling, of the modified deoxynucleoside 5-ethynyl-2'-deoxyuridine (EDU) incorporated into replicated DNA through incremental pulsing times. This methodology, which we designated as EdU-Coupled Fluorescence Intensity (E-CFI) analysis, can be applied to cell types with very distinct cell cycle features, and has shown excellent agreement with established techniques of cell cycle analysis. Useful modifications to the original protocol (Pereira et al., Oncotarget, 8:40514-40,532, 2017) have been introduced to increase flexibility in data collection and facilitate data analysis.
Topics: Cell Culture Techniques; Cell Cycle; Cell Line; Click Chemistry; DNA; DNA Replication; Deoxyuridine; Flow Cytometry; Humans
PubMed: 34085222
DOI: 10.1007/978-1-0716-1538-6_12 -
Chemical Research in Toxicology Apr 2019After thymidine (dT) was treated with a Fenton-type reagent and further incubated for a long period (6 days) under physiological conditions (37 °C, pH 7.4), a new...
After thymidine (dT) was treated with a Fenton-type reagent and further incubated for a long period (6 days) under physiological conditions (37 °C, pH 7.4), a new product, named dT*, was detected by HPLC in addition to the free thymine base and the known oxidative dT damage, 5-formyl-2'-deoxyuridine (fdU). dT* was found to be formed from fdU. The structure of dT* was determined to be 3-amino-2-carbamoyl-2-propenal-N-2'-deoxyriboside, a pyrimidine ring-opened product from fdU, on the basis of H- and C NMR analyses and mass spectra. From the model compound 1-methyl-5-formyluracil, a similar ring-opened product was formed after the incubation. dT* was also detected in DNA treated with a Fenton-type reagent or γ-rays, followed by the prolonged incubation. dT* will be a new promising marker of oxidative DNA damage. The possible role of this product in oxy-radical-induced mutagenesis is discussed.
Topics: Animals; Cattle; DNA; DNA Damage; Deoxyuridine; Molecular Structure; Oxidation-Reduction; Pyrimidines
PubMed: 30785277
DOI: 10.1021/acs.chemrestox.8b00401 -
Organic & Biomolecular Chemistry Apr 2015The representative DNA-labeling agent 5-ethynyl-2'-deoxyuridine (EdU) was chemically modified to improve its function. Chemical monophosphorylation was expected to...
The representative DNA-labeling agent 5-ethynyl-2'-deoxyuridine (EdU) was chemically modified to improve its function. Chemical monophosphorylation was expected to enhance the efficiency of the substrate in DNA polymerization by circumventing the enzymatic monophosphorylation step that consumes energy. In addition, to enhance cell permeability, the phosphates were protected with bis-pivaloyloxymethyl that is stable in buffer and plasma, and degradable inside various cell types. The phosphorylated EdU (PEdU) was less toxic than EdU, and had the same or a slightly higher DNA-labeling ability in vitro. PEdU was also successfully applied to DNA labeling in vivo. In conclusion, PEdU can be used as a less toxic DNA-labeling agent for studies that require long-term cell survival or very sensitive cell lines.
Topics: 3T3 Cells; Animals; DNA; Deoxyuridine; HeLa Cells; Humans; Mice; Phosphorylation; Staining and Labeling
PubMed: 25777799
DOI: 10.1039/c5ob00199d -
Antimicrobial Agents and Chemotherapy Aug 1979The pharmacokinetics of the newly developed anti-herpes agent, E-5-(2-bromovinyl)-2'-deoxyuridine, was compared with that of the standard anti-herpes drug... (Comparative Study)
Comparative Study
The pharmacokinetics of the newly developed anti-herpes agent, E-5-(2-bromovinyl)-2'-deoxyuridine, was compared with that of the standard anti-herpes drug 5-iodo-2'-deoxyuridine. Both compounds were administered to mice at 100 mg/kg by either the intraperitoneal, subcutaneous, or oral route. The active blood drug levels achieved by E-5-(2-bromovinyl)-2'-deoxyuridine were considerably higher than those attained by 5-iodo-2'-deoxyuridine (serum peak concentrations: 40 to 100 and 4 to 10 mug/ml, respectively). Active blood drug levels could still be found 320 min after oral administration of E-5-(2-bromovinyl)-2'-deoxyuridine.
Topics: Animals; Antiviral Agents; Brain Chemistry; Bromodeoxyuridine; Floxuridine; Idoxuridine; Kinetics; Liver; Lung; Mice; Simplexvirus
PubMed: 225987
DOI: 10.1128/AAC.16.2.234 -
Acta Crystallographica. Section C,... Aug 1994The furanose ring in C10H12N2O5 adopts the O(4')-endo envelope conformation (0E) and the glycosidic torsion angle C(2)--N(1)--C(1')--O(4'), chi, is 245.2 (3) degrees....
The furanose ring in C10H12N2O5 adopts the O(4')-endo envelope conformation (0E) and the glycosidic torsion angle C(2)--N(1)--C(1')--O(4'), chi, is 245.2 (3) degrees. The pseudo-rotational parameters are P = 102.7 degrees and tau m = 5.2 degrees. The CH2OH group on C(5') has the t conformation [gamma = 179.2 (2) degrees].
Topics: Antiviral Agents; Deoxyuridine; Molecular Conformation; Molecular Structure; Structure-Activity Relationship; X-Ray Diffraction
PubMed: 7946150
DOI: 10.1107/s0108270193013381 -
European Journal of Medicinal Chemistry Jun 2016New aromatic and aliphatic 3'-O-acyl-5-fluoro-2'-deoxyuridine derivatives were synthesized and evaluated as candidates for prodrugs against various cancer cell lines. As...
New aromatic and aliphatic 3'-O-acyl-5-fluoro-2'-deoxyuridine derivatives were synthesized and evaluated as candidates for prodrugs against various cancer cell lines. As the most promising candidate for antimalignant therapeutics was found a dual-acting acyl derivative 7h, which apparently released not only the known anticancer nucleoside, 5-fluoro-2'-deoxyuridine (FdU), but also an additional active metabolite, acetylsalicylic acid, reinforcing thus therapeutic effect of FdU. Promising therapeutic indices showed also some aromatic dicarboxylic acids derivatives decorated with FdU esters (11 and 12).
Topics: Antineoplastic Agents; Cell Proliferation; Cell Survival; Cells, Cultured; Deoxyuridine; Dose-Response Relationship, Drug; Drug Screening Assays, Antitumor; Humans; Molecular Structure; Prodrugs; Structure-Activity Relationship
PubMed: 26994842
DOI: 10.1016/j.ejmech.2016.03.010 -
Journal of Medicinal Chemistry Oct 1982Results are described that demonstrate that the mechanism of action of the potent cytotoxic agent 5-nitro-2'-deoxyuridine (NO2-dUrd) involves thymidine (dThd) kinase...
Results are described that demonstrate that the mechanism of action of the potent cytotoxic agent 5-nitro-2'-deoxyuridine (NO2-dUrd) involves thymidine (dThd) kinase catalyzed formation of 5-nitro-2'-deoxyuridylate (NO2dUMP) and subsequent potent inhibition of thymidylate (dTMP) synthetase by this compound. The evidence for this is as follows: (a) cells lacking dThd kinase are not inhibited by high concentrations of NO2dUrd; (b) the drug has no effect on dThd or uridine (Urd) incorporation into nucleic acids but prevents incorporation of deoxyuridine (dUrd); (c) growth inhibition is reversed by dThd but not by dUrd; (d) NO2dUrd causes changes in deoxynucleoside triphosphate pool sizes which are characteristic of specific inhibition of dTMP synthetase; (e) cells treated with [3H]NO2dUrd possess macromolecular bound [3H]NO2dUMP, which has properties characteristic of the NO2dUMP-dTMP synthetase complex. Treatment of L1210 leukemic mice at 400 mg/kg daily for 6 days gave only a 33% increase in life span, probably because of its rapid degradation to the inactive nitrouracil.
Topics: Adenosine Triphosphate; Animals; Antineoplastic Agents; Cells, Cultured; DNA; Deoxyuridine; Female; Leukemia L1210; Mice; RNA; Time Factors
PubMed: 6183429
DOI: 10.1021/jm00352a033 -
Gut Microbes 2023The composition of the intestinal bacterial community is well described, but recent research suggests that the metabolism of these bacteria plays a larger role in health...
The composition of the intestinal bacterial community is well described, but recent research suggests that the metabolism of these bacteria plays a larger role in health than which species are present. One fundamental aspect of gut bacterial metabolism that remains understudied is bacterial replication. Indeed, there exist few techniques which can identify actively replicating gut bacteria. In this study, we aimed to address this gap by adapting 5-ethynyl-2'-deoxyuridine (EdU) click chemistry (EdU-click), a metabolic labeling method, coupled with fluorescence-activated cell sorting and sequencing (FACS-Seq) to characterize replicating gut bacteria. We first used EdU-click with human gut bacterial isolates and show that many of them are amenable to this technique. We then optimized EdU-click and FACS-Seq for murine fecal bacteria and reveal that UCG-001 and are enriched in the replicating fraction. Finally, we labeled the actively replicating murine gut bacteria during exposure to cell wall-specific antibiotics . We show that regardless of the antibiotic used, the actively replicating bacteria largely consist of , suggesting the resistance of this taxon to perturbations. Overall, we demonstrate how combining EdU-click and FACSeq can identify the actively replicating gut bacteria and their link with the composition of the whole community in both homeostatic and perturbed conditions. This technique will be instrumental in elucidating bacterial replication dynamics in a variety of other ecological states, including colonization and species invasion, as well as for investigating the relationship between the replication and abundance of bacteria in complex communities.
Topics: Humans; Mice; Animals; Click Chemistry; Gastrointestinal Microbiome; Deoxyuridine; Bacteria
PubMed: 36823031
DOI: 10.1080/19490976.2023.2180317