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91.
Transduction with recombinant adeno-associated virus for gene therapy is limited by leading-strand synthesis. 总被引:36,自引:19,他引:17 下载免费PDF全文
K J Fisher G P Gao M D Weitzman R DeMatteo J F Burda J M Wilson 《Journal of virology》1996,70(1):520-532
Adeno-associated virus is an integrating DNA parvovirus with the potential to be an important vehicle for somatic gene therapy. A potential barrier, however, is the low transduction efficiencies of recombinant adeno-associated virus (rAAV) vectors. We show in this report that adenovirus dramatically enhances rAAV transduction in vitro in a way that is dependent on expression of early region 1 and 4 (E1 and E4, respectively) genes and directly proportional to the appearance of double-stranded replicative forms of the rAAV genome. Expression of the open reading frame 6 protein from E4 in the absence of E1 accomplished a similar but attenuated effect. The helper activity of adenovirus E1 and E4 for rAAV gene transfer was similarly demonstrated in vivo by using murine models of liver- and lung-directed gene therapy. Our data indicate that conversion of a single-stranded rAAV genome to a duplex intermediate limits transduction and usefulness for gene therapy. 相似文献
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Dariusz Latowski Jerzy Kruk Kvetoslava Burda Marta Skrzynecka-Jaskier Anna Kostecka-Guga?a Kazimierz Strza?ka 《European journal of biochemistry》2002,269(18):4656-4665
This paper describes violaxanthin de-epoxidation in model lipid bilayers. Unilamellar egg yolk phosphatidylcholine (PtdCho) vesicles supplemented with monogalactosyldiacylglycerol were found to be a suitable system for studying this reaction. Such a system resembles more the native thylakoid membrane and offers better possibilities for studying kinetics and factors controlling de-epoxidation of violaxanthin than a system composed only ofmonogalactosyldiacylglycerol and is commonly used in xanthophyll cycle studies. The activity of violaxanthin de-epoxidase (VDE) strongly depended on the ratio of monogalactosyldiacylglycerol to PtdCho in liposomes. The mathematical model of violaxanthin de-epoxidation was applied to calculate the probability of violaxanthin to zeaxanthin conversion at different phases of de-epoxidation reactions. Measurements of deepoxidation rate and EPR-spin label study at different temperatures revealed that dynamic properties of the membrane are important factors that might control conversion of violaxanthin to antheraxanthin. A model of the molecular mechanism of violaxanthin de-epoxidation where the reversed hexagonal structures (mainly created by monogalactosyldiacylglycerol) are assumed to be required for violaxanthin conversion to zeaxanthin is proposed. The presence of monogalactosyldiacylglycerol reversed hexagonal phase was detected in the PtdCho/monogalactosyldiacylglycerol liposomes membrane by 31P-NMR studies. The availability of violaxanthin for de-epoxidation is a diffusion-dependent process controlled by membrane fluidity. The significance of the presented results for understanding themechanism of violaxanthin de-epoxidation in native thylakoid membranes is discussed. 相似文献
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Marcus Müller Bodo Laube Hynek Burda Volkmar Bruns 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1992,171(4):469-476
Summary The cochlea of the mole rat Cryptomys hottentotus was investigated with physiological and anatomical methods. In order to reveal the place-frequency map of the cochlea, iontophoretic HRP-applications were made in the cochlear nucleus at physiologically characterized locations. Subsequent HRP-transport in auditory nerve fibres and labeling patterns of spiral ganglion cells within the cochlea were evaluated.A cochlear place-frequency map was constructed from 17 HRP-applications in the cochlear nucleus at positions where neurons had characteristic frequencies between 0.1 and 12.6 kHz. As in other mammals, high frequencies were found to be represented at the cochlear base, low frequencies at the cochlear apex. The placefrequency map had three distinct parts which were characterized by their different slopes. A clear overrepresentation of the frequencies between 0.6 and 1 kHz was revealed, in this frequency range the slope of the place-frequency map amounted to 5.3 mm/octave. As calculated from the regression analysis, below 0.6 kHz the slope of the cochlear place-frequency map amounted to 0.24 mm/octave, above 1 kHz to 0.9 mm/octave.As in other mammals width of the basilar membrane (BM) increased from the cochlear base towards the cochlear apex. Also in concordance with the findings in other mammals, BM-thickness decreased from the cochlear base to the apex. However, it was remarkable to find that there was no or little change in BM-width and thickness between 40 and 85% BM-length. It was also revealed that scala tympani was only 1/10th the size found in the rat or other mammals of similar body size.On the basis of the cochlear place-frequency map and the morphological findings we speculate that in Cryptomys hottentotus an acoustic fovea is present in the frequency range between 0.6 and 1 kHz. In analogy to echolocating bats, about half of the cochlea is devoted to the analysis of a narrow frequency band within the hearing range.Abbreviations
BM
basilar membrane
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CF
characteristic frequency
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CN
cochlear nucleus 相似文献
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Described: basic characteristics of reproduction and pre- and postnatal growth rate and development in Zambian mole-rats, Crypromys hottentotus (Bathyergidae), and compared with the available data on other small subterranean hystricognathous rodents. It is demonstrated that C. hottentotus from relatively rnesic habitats are monoamous and eusocial. The animals in each colony may be divided into several cateories according to their body size and pelage colouration (polymorhim) and prevailing beiavioural pattern (polyethism). Particularly breeding animals are sexually dimorphic. Freuent sexual interactions can be noted in the breeding pair even outside the female's estrus. Growth and sexual behaviour are pheromonally suppressed in the offspring (subordinate animals by their mutual contact and, particularly, by the parents (dominant animals). Most eflicient supression of growth is accomplished by a lactating female. It is arued that eusociality of the naked mole-rat, Heterocephalus glaber, is not qualitatively unique and has not evolved as an immediate response to aridity and dispersed food resources as generally considered. Evidence is presented that eusociality may have evolved as a direct consequence of decreasing the body size combined with retention of a long developmental time (phylogenetic constraint), limitations to store fat reserves, reduction of female's activit and consequent constraints of prenancy. Gestation had to be shortened at the cost of bearing altricial young and prolongation of tie postnatal development. Monogamy and establishment of a caste of helpers may be understood as energy-saving mechanisms of breeding females. Eusociality may be useful for survival in arid habitats, yet reasons for origin should te distinguished from the current (manifold) utility. 相似文献
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Jaroslav A. Hubacek Silvie Bloudickova Ruzena Kubinova Hynek Pikhart Ondrej Viklicky Martin Bobak 《Biochemical genetics》2009,47(9-10):688-693
A possible association between end-stage renal disease (ESRD) and apolipoprotein E (APOE) polymorphism was found in some but not all studies. We have analyzed the APOE genotypes in 995 hemodialyzed patients (cases) and a sample of 6242 healthy individuals (controls) in the Czech Republic. There was a statistically significant difference in the frequency of APOE alleles between cases and controls, with more carriers of the APOE2 allele in ESRD patients (15.9%) than in controls (12.2%) (P = 0.005). The odds ratio of ESRD for the APOE2 allele, compared with APOE3E3 homozygotes, was 1.37 (95% confidence interval 1.13–1.67). The strength of the association increased with the time spent on hemodialysis: the odds ratio of all-cause ESRD in patients dialyzed for eight or more years was 1.27 (0.94–1.71), for 1–8 years 1.41 (1.09–1.81), and less than 1 year (nonsurvivors) 1.94 (0.88–4.18). This study suggests that the APOE2 allele is a possible genetic risk factor for all-cause ESRD in Caucasians. 相似文献