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1.
Abstract 5-Ethyl-2′-deoxyurldine (EDU) is phosphorylated to a much greater extent by herpes simplex virus (HSV)-infected Vero cells than by mock-infected cells. Within the infected cells, EDU is preferentially incorporated into viral DNA and more inhibitory to viral than cellular DNA synthesis 相似文献
2.
《Analytical biochemistry》1986,154(2):436-440
We describe time-resolved fluoroimmunoassay of 5-methyl-2′-deoxycytidine (5MedCyd). The assay is based on the use of a highly specific antiserum raised in rabbits against BSA-conjugated 5-methylcytidine (5MeCyd). The tracer in the solid-phase time-resolved fluoroimmunoassay (TR-FIA) was antigen-selected anti-5MedCyd labeled with Europium. Thyroglobulin-linked 5MeCyd served as the solid-phase antigen. The measuring range for the fluoroimmunoassay was from less than 1 to 5000 pmol per assay of 5MedCyd. A good correlation between the results obtained with the TR-FIA and HPLC was demonstrated when the methods were applied to the measurement of methylation in human leukemic cells and other DNA samples. TR-FIA has several advantages over the more laborious techniques available so far: (i) high sensitivity, (ii) large assay ranges, (iii) rapidity and large number of simultaneous assays, (iv) simplicity, and (v) low cost provided that the laboratory has equipment for time-resolved fluorometry. 相似文献
3.
Anastassia Kanavarioti 《Origins of life and evolution of the biosphere》1986,17(1):85-103
We have studied the hydrolysis of guanosine 5-phospho-2-methylimidazolide, 2-MeImpG, in aqueous buffered solutions of various pH's at 75°C and 37°C. At 75°C and pH1.0, two kinetic processes were observed spectrophotometrically: the first and more rapid one is attributed to the hydrolysis of the phosphoimidazolide P-N bond; the second and much slower one, to the cleavage of the glycosidic bond. At 37°C, pH 2.0, the spectrophotometrically determined rate constant of P–N bond hydrolysis was confirmed by using high pressure liquid chromatography, HPLC. With the latter technique it was possible to separate reactants and products and also to extend the pH-rate profile into the neutral region where rates are slower and, therefore, difficult to measure spectrophotometrically. The pH-rate profiles at both temperatures exhibit similar behavior. At pH<2 the pseudo-first-order rate constant increases with decreasing pH; in the region 27. These data are consistent with a reactivity order zwitterion>anion for P–N bond hydrolysis. It is noteworthy that P–N bond hydrolysis in phosphoimidazolides is very slow compared to other phosphoramidates. This may be one of the reasons why this compound showed extraordinary ability in forming long oligomers under template-directed conditions. 相似文献
4.
Shan-Shan Gong Jian Sun Yue-Hai You Ji-Zong Chen Guo-Dong Liu 《Nucleosides, nucleotides & nucleic acids》2016,35(6):295-304
An efficient P(V)–N activation method for the synthesis of 5-carboxy-2′-deoxyuridine and 5-carboxy-2′-deoxycytidine triphosphates directly from the corresponding phosphoropiperidate precursors has been developed. 相似文献
5.
P. P. Torrence D. Alster S. Huss G. Gosaelin J.-L. Imbach 《Nucleosides, nucleotides & nucleic acids》2013,32(1-2):521-522
Abstract Sequential substitution of xyloadenosine into the trimeric and tetrameric 2–5A cores1 allows evaluation of the importance of the 3′ hydroxyl groups to 2′5′-phosphodiesterase (PDE) activity. 相似文献
6.
Krystyna Lesiak Bogdan Uznanski Paul F. Terrence 《Nucleosides, nucleotides & nucleic acids》2013,32(5-6):1055-1056
Abstract 2′,5′-Oligoadenylate 5′-triphosphates (2-5A) as products of 2-5A synthetase and activators of ribonuclease L (RNase L), are mediators in one of the mechanisms of interferon′s antiviral action. Upon activation, RNase L inhibits protein synthesis due to the degradation of RNAs. This activity of 2-5A could possibly find an application in virus or cancer chemotherapy, but two major barriers prevent the use of 2′,5′-linked oligoadenylates as therapeutic agents. The 2-5A is readily degraded by a 2′,5′ phosphodiesterase and as a highly negatively charged molecule, is not readily taken up by cells. One possible solution to this latter limitation might be found in chemical modifications of the 2-5A structure. Many analogues of 2-5A have been already obtained with modified base, ribose or phosphate moieties. While these have provided some important information about the enzyme- activator interactions, the cell permeability problem still remains unsolved. One of the major obstacles in this study is lack of a convenient method of synthesis of 2′,5′ ribonucleotides of widely varying structure. 相似文献
7.
Igor A. Mikhailopoulo Elena N. Kalinichenko Tatjana L. Podkopaeva Thomas Wenzel Helmut Rosemeyer Frank L. Seela 《Nucleosides, nucleotides & nucleic acids》2013,32(1-3):445-464
Abstract Synthesis of (2′ → 5′)ApApA analogues containing 1-deazaadenosine at different positions is described (32–34). The approach used the phosphotrieer methodology in solution and utilized 3′-O-benzoylated derivatives of the N6-protected 5′-O-monomethoxytrityl-1-deazaadenosine as starting material. 相似文献
8.
Danny P. C. McGee Chandra Vargeese Yansheng Zhai Gary P. Kirschenheuter Alecia Settle Colleen R. Siedem 《Nucleosides, nucleotides & nucleic acids》2013,32(6):1329-1339
Abstract The synthesis of 2′-amino-2′-deoxypyrimidine 5′-triphosphates is described. The 2′-amino-2′-deoxyuridine 5′-triphosphate is obtained from uridine in four steps with 25% overall yield. The 2′-amino-2′-deoxycytidine 5′-triphosphate is obtained from uridine in seven steps with 13% overall yield. 相似文献
9.
Jun-Ichi Yamashita Hiroshi Matsumoto Kazuhiro Kobayashi Kazuharu Noguchi Mitsugi Yasumoto Tohru Ueda 《Nucleosides, nucleotides & nucleic acids》2013,32(5-6):1145-1146
Abstract A practical synthesis of 3′-O-benzyl-2′-deoxy-5-trifluoromethyluridine was established which involves a selective 3′-O-benzylation of 2′-deoxy-5′-O-trityl-5-iodouridine followed by a cross-coupling with trifluoromethylcopper. 相似文献
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11.
Abstract The use of the Fmoc group for the protection of the hydroxy functions of the sugar moiety gave an improved overall yield of 2′-deoxy-5-azacytidine (6β), due to the mildly-basic conditions required for its removal from the protected nucleoside. 相似文献
12.
Bruce S. Ross Mingming Han Vasulinga T. Ravikumar 《Nucleosides, nucleotides & nucleic acids》2013,32(7):765-770
An efficient process to synthesize 5′-O-dimethoxytrityl-N4-benzoyl-5-methyl-2 ′-deoxycytidine in high yield and quality is described. Final benzoylation was improved by developing a method to selectively hydrolyze benzoyl ester impurities. This inexpensive approach was scaled up to multi-kilogram quantities for routine use in oligonucleotide therapeutics. 相似文献
13.
G. F. Jirikowski J. F. Ramalho-Ortigao T. Lindl H. Seliger 《Histochemistry and cell biology》1989,91(1):51-53
Summary A synthetic oligonucleotide probe, complementary to oxytocin m-RNA was labelled enzymatically with 5-bromo-2-deoxyuridine (5-BrdU) and, with [-32P]-ATP. The labelled probes were used for in situ, hybridization of histological sections of the mouse hypothalamus. A monoclonal antibody to 5-BrdU and the streptavidine-peroxidase technique were used in order to visualize hybridization with the 5-BrdU labelled probe. In situ hybridization with [32P] labelling was detected autoradiographically. With both methods hybridized neurons were visible in the magnocellular hypothalamic nuclei. While immunostaining and radio-labelling provided similar localization of oxytocin m-RNA, only the immunocytochemical technique showed clear cellular resolution of the reaction product. In situ hybridization with 5-BrdU labelled probes followed by 5-BrdU immunocytochemistry seems to be a powerful alternative to common autoradiographic techniques. 相似文献
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16.
《Nucleosides, nucleotides & nucleic acids》2013,32(12):2093-2104
Abstract A summary delineating the large scale synthetic studies to prepare labeled precursors of ribonucleosides-3′,4′,5′,5″- 2H 4 and -2′,3′,4′,5′,5″- 2H 5 from D-glucose is presented. The recycling of deuterium-labeled by-products has been devised to give a high overall yield of the intermediates and an expedient protocol has been elaborated for the conversion of 3-O-benzyl-α,β-D-allofuranose-3,4-d 2 6 to 1-O-methyl-3-O-benzyl-2-O-t-butyldimethylsilyl-α,β-D-ribofuranose-3,4,5,5′-d 4 16 (precursor of ribonucleosides-3′,4′,5′,5″- 2H 4 ) or to 1-O-methyl-3,5-di-O-benzyl-α,β-D-ribofuranose-3,4,5,5′-d 4 18 (precursor of ribonucleosides-3′,4′,5′,5″- 2H 4 ). 相似文献
17.
Isolation of Chlamydia trachomatis by Use of 5-Iodo-2-Deoxyuridine—Treated Cells 总被引:8,自引:0,他引:8
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Irradiated McCoy cells have provided a useful technique for the isolation of Chlamydia trachomatis strains, among which are found the etiological agents of trachoma, inclusion conjunctivitis, and lymphogranuloma venereum. Because irradiation is not always readily available, 5-iodo-2-deoxyuridine (IUDR) treatment of cells was investigated as a substitute procedure. IUDR-treated cells were found to be as sensitive to C. trachomatis infection as were irradiated McCoy cells. Stock chlamydial strains gave similar titers of iodine-stained inclusions in either system. When cells treated with IUDR were compared with irradiated cells for the isolation of C. trachomatis from clinical specimens, 5 of 138 specimens yielded isolates in IUDR-treated cells not found in irradiated ones, and one isolate was obtained from irradiated but not from IUDR-treated cells. In those 56 cases where inclusions were seen in both systems, there were significantly more inclusions in IUDR-treated than in irradiated cells. Although this series of cultures is too small to determine whether IUDR-treated cells are superior to irradiated ones for the isolation of C. trachomatis, the data indicate that IUDR treatment is at least equally effective. 相似文献
18.
Two independently isolated 5-fluoro-2-deoxyuridine (FUdR)-resistant mutant lines of Arabidopsis thaliana (L.) Heynh., FUD-1 and FUD-2, were identified by screening M2 populations of ethylmethane-sulfonatemutagenized seeds. The resistance was found to be due to single, recessive, nuclear gene mutations. Genetic complementation tests indicated that these two mutations were in the same gene locus, which was designated fur1, and mapped to linkage group four of Arabidopsis. Enzyme assays indicated that the mutants were not defective in thymidine-kinase activity. Greatly reduced concentrations of intracellular 3H were detected in fur1/fur1 plants compared with the wild type after incubation of wild-type and resistant plants in a medium with [3H]FUdR, indicating that either reduced uptake of FUdR or enhanced efflux of FUdR metabolites was the major reason for FUdR-resistance. fur1/fur1 plants also had significantly decreased uptake of thymidine and uridine compared with the wild type but no difference was found in the uptake of adenosine, guanosine, thymine, uracil or amino acids. It is suggested that the transport system affected in the fur1/fur1 mutants is one specific to pyrimidine nucleosides.Abbreviations BUdR
5-bromodeoxyuridine
- FdUMP
5-fluoro-2-deoxyuridine monophosphate
- FUdR
5-fluoro-2-deoxyuridine
- FUR
fluorouridine
- TK
thymidine kinase
- TS
thymidylate synthetase
We thank Dr. George W. Haughn (Department of Biology, University of Saskatchewan) for providing Arabidopsis line W100 and Dr. George Mourad (Department of Biology, University of Saskatchewan) for help and advice. This work was supported by a Research Grant from the Natural Sciences and Engineering Research Council of Canada to J.K. K.W. is grateful for a University of Saskatchewan Graduate Scholarship. 相似文献
19.
Wenlong Li 《Bioscience Hypotheses》2009,2(5):295-297
Type 2 diabetes is characterized by insulin resistance and β-cell dysfunction. The pathway of acylation-stimulating protein (ASP) and its specific receptor, C5a-like receptor 2 (C5L2), involves in the effective clearance of plasma glucose and free fat acid. Abnormal ASP-C5L2 pathway may induce insulin resistance, as well as cause hyperglycemia and elevated plasma free fat acid. High levels of plasma glucose and free fat acid induce β-cell apoptosis and dysfunction. We proposed that the abnormality of ASP-C5L2 pathway contributes to progression of type 2 diabetes. 相似文献