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21.
Synthesis and properties of oligodeoxynucleotides containing the analogue 2'-deoxy-4'-thiothymidine. 总被引:1,自引:1,他引:0 下载免费PDF全文
The 2'-deoxythymidine analogue 2'-deoxy-4'-thiothymidine has been incorporated, using standard methodology, into a series of dodecadeoxynucleotides containing the EcoRV restriction endonuclease recognition site (GATATC). The stability of these oligodeoxynucleotides and their ability to act as substrates for the restriction endonuclease and associated methylase have been compared with a normal unmodified oligodeoxynucleotide. No problems were encountered in the synthesis despite the presence of a potentially oxidisable sulfur atom in the sugar ring. The analogue had very little effect on the melting temperature of the self-complementary oligoeoxynucleotides so synthesised and all had a CD spectrum compatible with a B-DNA structure. The oligodeoxynucleotide containing one analogue in each strand within the recognition site, adjacent to the bond to be cleaved (i.e. GAXATC, where X is 2'-deoxy-4'-thiothymidine), was neither a substrate for the endonuclease nor was recognized by the associated methylase. When still within the recognition hexanucleotide but two further residues removed from the site of cleavage (i.e. GATAXC), the oligodeoxynucleotide was a poor substrate for both the endonuclease and methylase. Binding of the oligodeoxynucleotide to the endonuclease was unaffected but the kcat value was only 0.03% of the value obtained for the parent oligodeoxynucleotide. These results show that the incorporation of 2'-deoxy-4'-thionucleosides into synthetic oligodeoxynucleotides may shed light on subtle interactions between proteins and their normal substrates and may also show why 2'-deoxy-4'-thiothymidine itself is so toxic in cell culture. 相似文献
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OBJECTIVES--In an epidemic: to measure the incidence and risk of complications of influenza; to determine the effect of pre-existing disease on complications; to estimate vaccine uptake and efficacy. DESIGN--Case-control study. SETTING--Primary care: two group practices. SUBJECTS--342 of the 395 cases of clinically diagnosed influenza reported to the general practice surveillance of infectious diseases scheme of the Public Health Laboratory Service during the 1989 epidemic, and 342 age and sex matched controls. INTERVENTIONS--Examination of records. MAIN OUTCOME MEASURES--Documented recognised complications; hospital admission; previous vaccination. RESULTS--Of 15 recognised complications, bronchitis was the commonest (rate 190.1/1000 cases) and significantly commoner in cases (summary odds ratio 9.7) after adjusting for higher consultation rates (mean 6.1 per annum v 4.2 among controls; p < 0.0001). No deaths were recorded. The risk of bronchitis complicating influenza was higher in patients with pre-existing illnesses regarded as an indication for vaccination (odds ratio 3.3; p < 0.0001). Observed vaccination efficacy in those with pre-existing illnesses and in elderly subjects was high (63% and 77% respectively) but uptake was low (4.5% and 6.1% respectively). CONCLUSIONS--Bronchitis complicates about one fifth of all cases of influenza presenting to general practitioners. Patients with pre-existing illnesses regarded as an indication for vaccination are particularly at risk. Vaccine uptake is extremely low, precluding an unequivocal demonstration of a protective effect. 相似文献
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We examined variation in courtship activity and hormone levels of male and female wood ducks (Aix sponsa) in relation to hatch date. Young wood ducks were assigned in groups of 8 (4 males and 4 females) to 4 experimental pens; 2 pens contained early-hatched ducks (3-12 April) and 2 pens contained late-hatched ducks (7-16 June). Courtship behaviors occurred less frequently in October and November than in December and January-February for both early- and late-hatched groups. Early-hatched wood ducks participated in courtship more frequently and formed pair bonds sooner than late-hatched individuals. Testosterone (T) and androstenedione (AD) levels of males did not differ between treatment groups; however, average levels of luteinizing hormone (LH) were greater for early-hatched males. Differences in LH occurred in 2 of 12 samples (6 October and 3 November); otherwise, LH did not vary by treatment. Levels of T and LH in males varied temporally, but there was no significant temporal variation in AE levels. Temporal variation in hormone levels was similar for early- and late-hatched males. Estradiol (E), progesterone (P), and LH levels of females did not differ between treatment groups. There was temporal variation in levels of E and LH, but not of P; this variation was similar for early- and late-hatched females. These data indicate that behavioral differences occurring temporally and between early- and late-hatched wood ducks were not related to corresponding differences in hormone levels. 相似文献
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Effects of a variety of cytokines and inducing agents on vascular permeability factor mRNA levels in U937 cells. 总被引:7,自引:0,他引:7
Vascular permeability factor (VPF) is an approximately 40-kDa disulfide-linked dimeric glycoprotein that is active in increasing blood vessel permeability, endothelial cell growth and angiogenesis. Little is known about VPF gene regulation. In this study, we investigated the effects of a variety of cytokines and inducing agents on VPF mRNA levels in the monocyte-like U937 cell line. Transforming growth factor-beta 1 caused a 1.8-fold increase in VPF mRNA levels after 4 hours, followed by a decline to basal levels by 18 hours. Phorbol 12-myristate 13-acetate, a potent inducer of the differentiation of U937 cells, caused a 12.5-fold increase in VPF mRNA levels at 24 hours, coinciding with the differentiation of these monocyte-like cells into macrophage-like cells. 相似文献
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John Finnan Joanne Fitzgerald John Connolly Alison Donnelly 《Global Change Biology Bioenergy》2012,4(3):311-329
Renewable energy and greenhouse gas (GHG) reduction targets are driving an acceleration in the use of bioenergy resources. The environmental impact of national and regional development plans must be assessed in compliance with the EU Strategic Environmental Assessment (SEA) Directive (2001/42/EC). Here, we quantify the environmental impact of an Irish Government bioenergy plan to replace 30% of peat used in three peat‐burning power stations, located within the midlands region, with biomass. Four plan alternatives for supplying biomass to the power plant were considered in this study: (1) importation of palm kernel shell from south‐east Asia, (2) importation of olive cake pellets from Spain and (3) growing either willow or (4) Miscanthus in the vicinity of the power stations. The impact of each alternative on each of the environmental receptors proposed in the SEA Directive was first quantified before the data were normalized on either an Irish, regional or global scale. Positive environmental impacts were very small compared to the negative environmental impacts for each of the plan alternatives considered. Comparison of normalized indicator values confirmed that the adverse environmental consequences of each plan alternative are concentrated at the location where the biomass is produced. The analysis showed that the adverse environmental consequences of biomass importation are substantially greater than those associated with the use of willow and Miscanthus grown on former grassland. The use of olive cake pellets had a greater adverse environmental effect compared to the use of peat whereas replacement of peat with either willow or Miscanthus feedstocks led to a substantial reduction in environmental pressure. The proposed assessment framework combines the scope of SEA with the quantitative benefits of life cycle assessment and can be used to evaluate the environmental consequences of bioenergy plans. 相似文献
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O'Rourke SM Yochem J Connolly AA Price MH Carter L Lowry JB Turnbull DW Kamps-Hughes N Stiffler N Miller MR Johnson EA Bowerman B 《Genetics》2011,189(3):767-778
Forward genetic screens provide a powerful approach for inferring gene function on the basis of the phenotypes associated with mutated genes. However, determining the causal mutation by traditional mapping and candidate gene sequencing is often the rate-limiting step, especially when analyzing many mutants. We report two genomic approaches for more rapidly determining the identity of the affected genes in Caenorhabditis elegans mutants. First, we report our use of restriction site-associated DNA (RAD) polymorphism markers for rapidly mapping mutations after chemical mutagenesis and mutant isolation. Second, we describe our use of genomic interval pull-down sequencing (GIPS) to selectively capture and sequence megabase-sized portions of a mutant genome. Together, these two methods provide a rapid and cost-effective approach for positional cloning of C. elegans mutant loci, and are also applicable to other genetic model systems. 相似文献