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1.
Introduction of 5'-terminal functional groups into synthetic oligonucleotides for selective immobilization 总被引:3,自引:0,他引:3
Oligodeoxyribonucleotides terminating in a 5'-primary amine group are synthesized using solid-phase supported phosphoramidite chemistry. The 5'-terminal amine group in the deprotected oligomers is further derivatized with either succinic anhydride to give 5'-carboxylic acid or with dithiobis(succinimidylpropionate) followed by treatment with dithioerythritol to produce 5'-thiol-terminated oligonucleotides. The 5'-thiol-terminated oligonucleotides are selectively immobilized on solid supports containing either p-chloromercuribenzoate or 2,2'-dithiobis(5-nitropyridine) activated thiol groups. 相似文献
2.
Negative-ion fast atom bombardment mass spectrometry is quite useful for the identification of products and by-products formed during the synthesis of nucleoside 3'-phosphoramidites. The data show that detritylation and oxidation are side reactions which occur during the synthesis of monomeric units used in the construction of oligodeoxyribonucleotides by the phosphite triester method. 相似文献
3.
Gregory A. Osenga Bruce C. Coull 《Journal of experimental marine biology and ecology》1983,67(3):221-225
Meiofauna were collected from adjacent areas with and without Spartina alterniflora Loisel culms. Environmental complexity resulting from root structure cannot be determined by simply observing S. alterniflora culm distribution. There were no significant differences between the two areas with respect to root density or root sizes; however, the number of live roots was significantly higher in the area containing Spartina culms. Nematodes, the most abundant taxon were positively correlated with live root density. In contrast, the number of dead roots and harpacticoid copepods were higher in the area without culms. This study suggests that nematodes are positively associated with micro-oxygenated zones produced by live S. alterniflora roots. Other measures of biogenic structure, such as total root density or root size fractions are not important in affecting nematode abundance. 相似文献
4.
Calcium content and distribution as a function of growth and transformation in the mouse 3T3 cell
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Total Ca content and that fraction of Ca sensitive to removal by the chelator ethylene glycol-bis(β-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA) have been investigated in the mouse 3T3 cell as a function of growth stage, transformation with SV40 virus, and serum levels of the media. Cells were allowed to grow through several doublings in media containing (45)Ca. The cellular content of (45)Ca was used to access total cell Ca. That fraction of (45)Ca removed by EGTA was presumed to represent primarily surface-localized Ca. The data are expressed on a per cell volume basis to compensate for size differences as a function of growth stage and transformation. During exponential growth phase, the 3T3 cell contains 525pmol Ca/μl cell volume. Of this, approx. 457 pmol/μl is not removable by EGTA and, presumably, is cytoplasmically located. This value is in close agreement with previous studies on the HeLa cell (470 pmol Ca/μl cell water after the removal of the surface Ca). The low level of EGTA- removable Ca present in the 3T3 cell during early exponential growth (68 pmol Ca/μl cell volume) increases progressively with increasing cell density, and upon quiescence it is sevenfold greater. In contrast, SV40- transformed 3T3 cells growing exponentially possess total levels of Ca which are approximately two-thirds the levels of the normal 3T3 cell. However, their EGTA-sensitive Ca is not significantly different from that of exponentially growing, normal 3T3 cells. As the transformed cells continue to grow at high density, their total ca and their sensitivity to EGTA do not change, in contrast to the normal 3T3 cell. Thus, an increase in Ca associated with the cell surface appears to be correlated with growth inhibition. This has been investigated further by regulating growth of the normal and transformed cell with alterations in the serum level of the media. In 4 percent calf serum the normal cell is stopped from continued proliferation. Growth stoppage under these conditions is characterized by a nearly fourfold increase in EGTA-removable Ca, similar to the increase observed upon quiescence in depleted 10 percent serum. Similar treatment of the transformed cell does not reduce its growth rate, nor does it significantly alter Ca distribution. However, at 0.5 percent medium serum levels, the SV40 3T3 growth rate is substantially reduced and, under these conditions, EGTA-removable Ca increases twofold. 相似文献
5.
Lai CM Yu MJ Brankov M Barnett NL Zhou X Redmond TM Narfstrom K Rakoczy PE 《Genetic vaccines and therapy》2004,2(1):3
Background
Leber's congenital amaurosis (LCA) is a severe form of retinal dystrophy. Mutations in the RPE65 gene, which is abundantly expressed in retinal pigment epithelial (RPE) cells, account for approximately 10–15% of LCA cases. In this study we used the high turnover, and rapid breeding and maturation time of the Rpe65 -/- knockout mice to assess the efficacy of using rAAV-mediated gene therapy to replace the disrupted RPE65 gene. The potential for rAAV-mediated gene treatment of LCA was then analyzed by determining the pattern of RPE65 expression, the physiological and histological effects that it produced, and any improvement in visual function. 相似文献6.
7.
Hierarchical modeling is becoming increasingly popular in epidemiology, particularly in air pollution studies. When potential confounding exists, a multilevel model yields better power to assess the independent effects of each predictor by gathering evidence across many sub-studies. If the predictors are measured with unknown error, bias can be expected in the individual substudies, and in the combined estimates of the second-stage model. We consider two alternative methods for estimating the independent effects of two predictors in a hierarchical model. We show both analytically and via simulation that one of these gives essentially unbiased estimates even in the presence of measurement error, at the price of a moderate reduction in power. The second avoids the potential for upward bias, at the price of a smaller reduction in power. Since measurement error is endemic in epidemiology, these approaches hold considerable potential. We illustrate the two methods by applying them to two air pollution studies. In the first, we re-analyze published data to show that the estimated effect of fine particles on daily deaths, independent of coarse particles, was downwardly biased by measurement error in the original analysis. The estimated effect of coarse particles becomes more protective using the new estimates. In the second example, we use published data on the association between airborne particles and daily deaths in 10 US cities to estimate the effect of gaseous air pollutants on daily deaths. The resulting effect size estimates were very small and the confidence intervals included zero. 相似文献
8.
9.
BACKGROUND: In the current study we compared the effects of temporal orienting of attention based on predictions carried by the intrinsic temporal structure of events (rhythm) and by instructive symbolic cues; and tested the degree of cognitive, strategic control that could be exerted over each type of temporal expectation. The experiments tested whether the distinction between exogenous and endogenous orienting made in spatial attention may extend to the temporal domain. TASK DESIGN AND MAIN RESULTS: In this task, a ball moved across the screen in discrete steps and disappeared temporarily under an occluding band. Participants were required to make a perceptual discrimination on the target upon its reappearance. The regularity of the speed (rhythmic cue) or colour (symbolic cue) of the moving stimulus could predict the exact time at which a target would reappear after a brief occlusion (valid trials) or provide no temporal information (neutral trials). The predictive nature of rhythmic and symbolic cues was manipulated factorially in a symmetrical and orthogonal fashion. To test for the effects of strategic control over temporal orienting based on rhythmic or symbolic cues, participants were instructed either to "attend-to-speed" (rhythm) or "attend-to-colour". Our results indicated that both rhythmic and symbolic (colour) cues speeded reaction times in an independent fashion. However, whilst the rhythmic cueing effects were impervious to instruction, the effects of symbolic cues were contingent on the instruction to attend to colour. FINAL CONCLUSIONS: Taken together, our results provide evidence for the existence of qualitatively separable types of temporal orienting of attention, akin to exogenous and endogenous mechanisms. 相似文献
10.