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101.
Farrow PJ Barrett LB Stevenson M Fisher KD Finn J Spice R Allan MA Berry M Logan A Seymour LW Read ML 《Nucleic acids research》2006,34(11):e80
Non-viral vectors are promising vehicles for gene therapy but delivery of plasmid DNA to post-mitotic cells is challenging as nuclear entry is particularly inefficient. We have developed and evaluated a hybrid mRNA/DNA system designed to bypass the nuclear barrier to transfection and facilitate cytoplasmic gene expression. This system, based on co-delivery of mRNA(A64) encoding for T7 RNA polymerase (T7 RNAP) with a T7-driven plasmid, produced between 10- and 2200-fold higher gene expression in primary dorsal root ganglion neuronal (DRGN) cultures isolated from Sprague–Dawley rats compared to a cytomegalovirus (CMV)-driven plasmid, and 30-fold greater expression than the enhanced T7-based autogene plasmid pR011. Cell-free assays and in vitro transfections highlighted the versatility of this system with small quantities of T7 RNAP mRNA required to mediate expression at levels that were significantly greater than with the T7-driven plasmid alone or supplemented with T7 RNAP protein. We have also characterized a number of parameters, such as mRNA structure, intracellular stability and persistence of each nucleic acid component that represent important factors in determining the transfection efficiency of this hybrid expression system. The results from this study demonstrate that co-delivery of mRNA is a promising strategy to yield increased expression with plasmid DNA, and represents an important step towards improving the capability of non-viral vectors to mediate efficient gene transfer in cell types, such as in DRGN, where the nuclear membrane is a significant barrier to transfection. 相似文献
102.
Because of the carcinogenicity of SV40 in rodents, and its possible distribution through the polio vaccine, many studies have been conducted to determine if there is an association between SV40 genomic infection and different types of cancer; sometimes, these studies included data on the prevalence of genomic infection in healthy subjects as secondary information. We reviewed all the studies that reported the prevalence of SV40 genomic infection in healthy subjects, tested by PCR based methods. The 20 articles considered here included 1103 samples from healthy subjects, with a prevalence of infection ranging from 0 to 25.6%, with high heterogeneity, and no association with the type of sample analyzed (Mantel-Haenszel OR: 0.74; 95% CI: 0.44-1.23). The wide variation in frequency pose problems in terms of study design; in fact, the representativeness of the samples used as controls in the published studies may be very limited. Larger studies on healthy subjects, tested for SV40 genomic infection at various genomic regions, conducted in different geographic areas, are needed. 相似文献
103.
The influence of melanin on radiation-induced bystander effects has been studied. Melanin is known to be a natural substance with proved radioprotective properties in different organisms and cell lines. It is non-toxic and is effective against acute and chronic irradiation. The lower the radiation dose, the higher the relative impact of melanin protection. In this study influence of melanin on human keratinocytes (HPV-G cells) has been studied using the colony-forming assay. We have shown that bystander donor medium from 0.5 Gy irradiated cells when transferred to unirradiated cells, caused almost the same effect as direct irradiation. Melanin increased the colony-forming ability of bystander recipient cells when it was added into culture medium before irradiation. The effect of melanin added after irradiation was to produce less protection in both the directly irradiated and bystander medium treated groups. The absorption spectrum of the filtered medium is identical to one of the intact culture medium showing that melanin was not present in filtered medium. Thus, it cannot protect recipient cells but reduces the amount of the bystander effect. It is concluded that melanin added before irradiation effectively decreased the radiation dose. The reduction of the impact of the bystander signal on recipient cells when melanin was added to the donor medium after harvest but before filtration, may mean that the bystander signal has a physical component as melanin can absorb all types of physical energy. 相似文献
104.
Ambulatory cardiology began in 1959 in a department of pathology to answer a question raised at the autopsy table: are high heart rates in apparently healthy individuals a risk factor for developing coronary artery disease? This question led to the development of a miniature monitor and a new kind of electrode, which enabled clinicians to measure EKG signals during activity and over prolonged periods of time. These electrodes are now used universally for diagnosis and for monitoring the heart during a myriad of different activities and circumstances. The story of the development of the monitor and electrodes illustrates the ways in which ideas and discovery lead to applications and advances in medicine. 相似文献
105.
Much of the current interest in pollen time series analysis is motivated by the possibility that pollen series arise from
low-dimensional chaotic systems. If this is the case, short-range prediction using nonlinear modeling is justified and would
produce high-quality forecasts that could be useful in providing pollen alerts to allergy sufferers. To date, contradictory
reports about the characterization of the dynamics of pollen series can be found in the literature. Pollen series have been
alternatively described as featuring and not featuring deterministic chaotic behavior. We showed that the choice of test for
detection of deterministic chaos in pollen series is difficult because pollen series exhibit power spectra. This is a characteristic that is also produced by colored noise series, which mimic deterministic chaos in
most tests. We proposed to apply the Ikeguchi–Aihara test to properly detect the presence of deterministic chaos in pollen
series. We examined the dynamics of cedar (Cryptomeria japonica) hourly pollen series by means of the Ikeguchi–Aihara test and concluded that these pollen series cannot be described as
low-dimensional deterministic chaos. Therefore, the application of low-dimensional chaotic deterministic models to the prediction
of short-range pollen concentration will not result in high-accuracy pollen forecasts even though these models may provide
useful forecasts for certain applications. We believe that our conclusion can be generalized to pollen series from other wind-pollinated
plant species, as wind speed, the forcing parameter of the pollen emission and transport, is best described as a nondeterministic
series that originates in the high dimensionality of the atmosphere. 相似文献
106.
Mitotic Recombination in Yeast: Isolation and Characterization of Mutants with Enhanced Spontaneous Mitotic Gene Conversion Rates 总被引:3,自引:2,他引:3 下载免费PDF全文
Semi-dominant mutants displaying greatly elevated (up to 200-fold above control) levels of spontaneous mitotic recombination have been isolated in a disomic haploid strain of yeast heteroallelic at the arg4 locus. They are designated by the symbol MIC. The mutants variously exhibit associated sensitivity to UV and ionizing radiation and to methyl methanesulfonate, enhanced UV-induced mitotic recombination, and enhanced spontaneous forward mutation rates. Possible enzyme defects and involvement in repair and editing of DNA are discussed. The mutants are expected to simplify the analysis of recombination pathways in yeast. 相似文献
107.
108.
Chapman NH Bonnet J Grivet L Lynn J Graham N Smith R Sun G Walley PG Poole M Causse M King GJ Baxter C Seymour GB 《Plant physiology》2012,159(4):1644-1657
109.
Bioinsecticides are important in the control of disease vectors, but data regarding their physiological effects on target insects are incomplete. This study describes morphological changes that occur in the midgut of third instar Aedes aegypti L. (Diptera: Culicidae) following treatment with a methanolic extract of Annona coriacea (Magnoliales: Annonaceae). Dissected midguts were subdivided into anterior and posterior regions and analyzed by light and scanning electron microscopy. Insects exposed to the extract displayed intense, destructive cytoplasmic vacuolization in columnar and regenerative midgut cells. The apical surfaces of columnar cells exhibited cytoplasmic protrusions oriented toward the lumen, suggesting that these cells could be involved in apocrine secretory processes and/or apoptosis. We report that A. coriacea extracts induced morphological alterations in the midgut of A. aegypti midgut larvae, supporting the use of plant extracts for control of the dengue vector. 相似文献
110.
Despite the worldwide distribution, toxicity and commercial, industrial and medical impacts jellyfish present, many aspects of their ecology remain poorly understood. Quantified here are important ecological parameters of Chironex fleckeri medusae, contributing not only to the understanding of an understudied taxon, the cubozoa, but also to the broader understanding of jellyfish ecology. C. fleckeri medusae were collected across seven seasons (1999, 2000, 2003, 2005-07 and 2010), with growth rates, temporal variation in the medusae season onset and differences in population structure between estuarine and coastal habitats quantified. With a mean of 2 September ± 2 d (mean ± 95% confidence limits), the earliest date of metamorphosis was temporally constrained between seasons, varying by only 7 d (30 August to 5 September). Juvenile medusae appeared to be added over an extended period, suggesting polyp metamorphosis was an ongoing process once it commenced. At a maximum of 3 ± 0.2 mm d(-1) IPD, medusae growth to an asymptotic size of ~190 mm IPD was rapid, yet, with the oldest medusae estimated to be ~78 d in age, medusae did not appear to accumulate along the coastline. Furthermore, a greater proportion of juveniles were observed along the coastline, with estuarine populations typified by larger medusae. With key aspects of C. fleckeri's ecology now quantified, medusae season management protocols can be further developed. 相似文献