共查询到20条相似文献,搜索用时 15 毫秒
1.
P. Laktionov J-E Dazard J. Piette E. Vives E. Rykova V. Vlassov 《Nucleosides, nucleotides & nucleic acids》2013,32(6-7):1697-1699
Abstract Oligonucleotides (ODNs) conjugated to rhodamin (Rh) and 4-[(N-2-chloroethyl-N-methyl)amino] benzylamine were used to investigate ODNs transport into keratinocytes. Affinity labeling of two proteins, 63 and 35 kDa, and the inhibition of the affinity labeling and ODNs uptake by the cells in the presence of nucleic acids, polyanions and trypsin suggest, that the proteins are involved in transport of nucleic acids in keratinocytes. 相似文献
2.
Ultrasound (US) has been found to facilitate the transport of DNA across cell membranes. However, the transfection efficiency is generally low, and the expression duration of the transfected gene is brief. In this study, a tertiary polycation, Poly(2-(dimethylamino) ethyl methacrylate) (PDMAEMA), was used as a carrier for US-mediated gene transfection. Its in-vitro and in-vivo effects on the transfection efficiency and the expression duration were evaluated. A mixture of pCI-neo-luc and PDMAEMA was transfected to cultured cells or mouse muscle by exposure to 1-MHz pulse US. A strong expression of luciferase was found 10 days after the transfection in vitro regardless of US exposure. However, effective transfection only occurred in the US groups in vivo. The transfection ability depended on the weight ratio of PDMAEMA to DNA, and was different for the in-vitro and in-vivo conditions. Lower weight ratios, e.g., 0.25, exhibited better in-vivo expression for at least 45 days. 相似文献
3.
A. Peyman H. Ragg T. Ulshöfer D. W. Will E. Uhlmann 《Nucleosides, nucleotides & nucleic acids》2013,32(7-9):1215-1219
Abstract The sequence-specificity of antisense oligonucleotides (ODN) against c-myc mRNA was tested by Northern blot analysis. Rat smooth muscle cells were treated with antisense or control ODN against c-myc modified by the “minimal protection strategy”. At 0.3 μM concentration the ODN show a very specific reduction in c-myc mRNA levels. Use of the “minimal protection strategy” minimizes nonspecific effects as observed for all-phosphorothioate ODN containing four consecutive guanine residues. 相似文献
4.
The use of synthetic oligonucleotides and their analogs to block gene expression by binding the complementary RNA sequences in cells, the antisense principle, has been limited by poor uptake of the agents by cells in culture. This review describes attempts to harness by chemical conjugation the ability of certain peptides that may cross membranes to enhance the cellular uptake of oligonucleotides. These include fusogenic and hydrophobic peptides, nuclear localization signals, receptor targeting and translocating peptides, and various combinations. We also outline briefly some popular methods of peptide–oligonucleotide conjugation. Finally, we review the use of noncovalent peptide additives and the recent studies of conjugates of peptide nucleic acid (PNA) with peptides. 相似文献
5.
Etienne van Bracht Luuk R. M. Versteegden Sarah Stolle Wouter P. R. Verdurmen Rob Woestenenk René Raavé Theo Hafmans Egbert Oosterwijk Roland Brock Toin H. van Kuppevelt Willeke F. Daamen 《PloS one》2014,9(11)
Lyophilisomes are a novel class of biodegradable proteinaceous nano/micrometer capsules with potential use as drug delivery carrier. Cell-penetrating peptides (CPPs) including the TAT peptide have been successfully implemented for intracellular delivery of a broad variety of cargos including various nanoparticulate pharmaceutical carriers. In the present study, lyophilisomes were modified using CPPs in order to achieve enhanced cellular uptake. Lyophilisomes were prepared by a freezing, annealing, and lyophilization method and a cystein-elongated TAT peptide was conjugated to the lyophilisomes using a heterobifunctional linker. Fluorescent-activated cell sorting (FACS) was utilized to acquire a lyophilisome population with a particle diameter smaller than 1000 nm. Cultured HeLa, OVCAR-3, Caco-2 and SKOV-3 cells were exposed to unmodified lyophilisomes and TAT-conjugated lyophilisomes and examined with FACS. HeLa cells were investigated in more detail using a trypan blue quenching assay, confocal microscopy, and transmission electron microscopy. TAT-conjugation strongly increased binding and cellular uptake of lyophilisomes in a time-dependent manner in vitro, as assessed by FACS. These results were confirmed by confocal microscopy. Transmission electron microscopy indicated rapid cellular uptake of TAT-conjugated lyophilisomes via phagocytosis and/or macropinocytosis. In conclusion, TAT-peptides conjugated to albumin-based lyophilisomes are able to enhance cellular uptake of lyophilisomes in HeLa cells. 相似文献
6.
Di Lu Yang An Simin Feng Xiaodan Li Aiping Fan Zheng Wang Yanjun Zhao 《AAPS PharmSciTech》2018,19(6):2610-2619
The complex design of multifunctional nanomedicine is beneficial to overcome the multiple biological barriers of drug delivery, but it also presents additional hurdles to clinical translation (e.g., scaling-up and quality control). To address this dilemma, we employed a simple imidazole-bearing polymer micelle for enhanced cellular uptake, facilitated endosomal escape, and on-demand release of a model drug, SN-38. The micelles were crosslinked by the reversible imidazole/Zn2+ coordination with a drug loading of ca. 4% (w/w) and a diameter less than 200 nm. Under mimicked tumor microenvironment (pH 6.8), the surface charge of micelles reversed from negative to positive, leading to enhanced micelles uptake by model 4T1 cells. Such effect was verified by fluorescent labelling of micelles. Compared to imidazole-free nanocarriers, the charge-reversal micelles delivered significantly more SN-38 to 4T1 cells. Due to the proton sponge effect, imidazole-bearing micelles could rapidly escape from endosomes compared to the control micelles, as evidenced by the kinetic analysis of micelle/endosome co-localization. The coordination crosslinking also enabled the acid-triggered drug release. This work provides a “three birds with one stone” approach to achieve the multifunctionality of nanocarriers without complicated particle design, and opens new avenues of advancing nanomedicine translation via simple tailored nanocarriers. 相似文献
7.
Valérie Hélin Marina Gottikh Zohar Mishal Frédéric Subra Claude Malvy Marc Lavignon 《Nucleosides, nucleotides & nucleic acids》2013,32(6-7):1721-1722
Abstract We studied the uptake and intracellular distribution of an FITC labelled phosphodiester oligodeoxynucleotide (ODN) vectorized by a dendrimeric structure in cell culture. 相似文献
8.
9.
Jean-Claude Lavoie Thérèse Rouleau Anita C. Truttmann Philippe Chessex 《Free radical research》2013,47(8):811-817
Background. In view of the functional capacity of glutathione synthesis in premature infants, and because the availability of cysteine is one the rate limiting steps in glutathione synthesis, we hypothesized that the low glutathione levels in premature infants may be due to immaturity of the active cellular uptake of cysteine.Objective. To document in cells from newborn infants the effect of maturity and gender on cysteine uptake and consequently on glutathione levels.Methods. Incorporation of L‐[35S] cysteine was measured in leukocytes from cord blood and from tracheal aspirates (TAC) of newborn infants of varying (gestational as well as postnatal) ages and gender. Cysteine uptake was correlated with glutathione in TAC.Results. The maturity of newborn girls positively influences cysteine uptake, which is responsible for 78% of the variation in their glutathione content. However, in newborn boys, gestational and postnatal ages did not influence the cysteine uptake.Discussion. Cysteine uptake appears to be the limiting step explaining the reported gender-related differences in glutathione as well as the low levels of this central antioxidant found in premature infants. The immature cysteine uptake found in cells from premature infants raises questions about the bioavailability of this conditionally essential amino acid in regimens of parenteral nutrition for human neonates. 相似文献
10.
Nikita Oskolkov Piret Arukuusk Dana-Maria Copolovici Staffan Lindberg Helerin Margus Kärt Padari Margus Pooga Ülo Langel 《International journal of peptide research and therapeutics》2011,17(2):147-157
Oligonucleotide-based gene regulation has a high potential in gene therapy, but the plasma membrane is impermeable for nucleic
acid polymers and, consequently, an efficient and non-toxic transfection agent is needed for their delivery into the cell.
In this study we present a novel series, NickFects, of chemically modified TP10 peptide-based delivery vectors used for the
cellular delivery of single-stranded oligonucleotides. These carriers, obtained by replacement of Ile8 by threonine in stearyl-TP10
and by modifying of tyrosine and/or threonine, respectively, by phosphorylation formed 300–500 nm in size peptide:oligonucleotide
nanocomplexes with negative surface charges. The highest splice-correcting effect was obtained when phosphorotiate 2′-O-methyl
oligonucleotides were transduced into cells by NickFect1 (NF1) or NickFect2 (NF2). In addition, we also show how a small modification
(one or two negative charges) in peptide sequence can affect its ability to deliver ONs into cells and increase their potency
in the splicing redirection assay. Our studies demonstrate that NF1 and NF2 have higher transfection efficacy for oligonucleotides
as compared to the most commonly used transfection agent Lipofectamine™ 2000 and lead to higher biological response in cells. 相似文献
11.
Wei-liang Ye Jiang-bo Du Bang-le Zhang Ren Na Yan-feng Song Qi-bing Mei Ming-gao Zhao Si-yuan Zhou 《PloS one》2014,9(5)
A PEG-based, folate mediated, active tumor targeting drug delivery system using DOX-hyd-PEG-FA nanoparticles (NPs) were prepared. DOX-hyd-PEG-FA NPs showed a significantly faster DOX release in pH 5.0 medium than in pH 7.4 medium. Compared with DOX-hyd-PEG NPs, DOX-hyd-PEG-FA NPs increased the intracellular accumulation of DOX and showed a DOX translocation from lysosomes to nucleus. The cytotoxicity of DOX-hyd-PEG-FA NPs on KB cells was much higher than that of free DOX, DOX-ami-PEG-FA NPs and DOX-hyd-PEG NPs. The cytotoxicity of DOX-hyd-PEG-FA NPs on KB cells was attenuated in the presence of exogenous folic acid. The IC50 of DOX-hyd-PEG-FA NPs and DOX-hyd-PEG NPs on A549 cells showed no significant difference. After DOX-hyd-PEG-FA NPs were intravenously administered, the amount of DOX distributed in tumor tissue was significantly increased, while the amount of DOX distributed in heart was greatly decreased as compared with free DOX. Compared with free DOX, NPs yielded improved survival rate, prolonged life span, delayed tumor growth and reduced the cardiotoxicity in tumor bearing mice model. These results indicated that the acid sensitivity, passive and active tumor targeting abilities were likely to act synergistically to enhance the drug delivery efficiency of DOX-hyd-PEG-FA NPs. Therefore, DOX-hyd-PEG-FA NPs are a promising drug delivery system for targeted cancer therapy. 相似文献
12.
Luigi E. Xodo Marianna Alunni-Fabbroni Gtorgio Manzini 《Nucleosides, nucleotides & nucleic acids》2013,32(7-9):1695-1698
Abstract The ability of (A,G)- and (G,T)-oligonucleotides to form triple-helices with a critical polypurine-polypyrimidine sequence of the c-Ki-ras promoter has been examined, together with their capacity to inhibit the expression of the chloramphenicol acetyl-transferase gene driven by the c-Ki-ras promoter in a transfected cellular system. 相似文献
13.
Maintenance of intracellular K+ concentrations that are not growth-limiting, in an environment of high Na+, is characteristic of NaCl-adapted cells of the glycophyte, tobacco (Nicotiana tabacum/gossii). These cells exhibited a substantially greater uptake of 86Rb+ (i.e. an indicator of K+) relative to unadapted cells. Potassium uptake into NaCl-adapted cells was 1.5-fold greater than unadapted cells at 0 NaCl and 3.5-fold greater when cells were exposed to 160 millimolar NaCl. The difference in net K+ uptake between unadapted and NaCl-adapted cells was due primarily to higher rates of entry rather than to reduced K+ leakage. Presumably, enhanced K+ uptake into adapted cells is a result of electrophoretic flux, and a component of uptake may be linked to vanadate-sensitive H+ extrusion. 相似文献
14.
C. Dell'aquila E. Vives J. L. Imbach B. Rayner 《Nucleosides, nucleotides & nucleic acids》2013,32(6-7):1683-1684
Abstract In this paper we described the synthesis on solid support of 5′-fluorescein labeled dodecathymidylates having interucleoside phosphodiester or phosphorothioate linkages temporary masked with enzymolabile S-pivaloyl-2-thioethyl (tBuSATE) groups. 相似文献
15.
C. Gosse A. S. Boutorine L. Jullien C. Hélène 《Nucleosides, nucleotides & nucleic acids》2013,32(6-7):1473-1476
Abstract The synthesis of a new fluorescently labeled medium-sensitive lipophilic oligonucleotide is reported. A fluorescent chalcone chromophore was introduced between the 5′ end of the nucleic acid and the fatty hydrocarbon chains. A blue shift of both absorption and emission wavelength maxima results from a transfer of the chromophore to a more hydrophobic medium or upon binding of the conjugate to unilamellar vesicles of egg phosphatidyl choline. These conjugates could be used as markers for cell uptake studies of lipophilic nucleic acid derivatives. 相似文献
16.
目的:研究bFGF反义硫代寡核苷酸增强肿瘤细胞对化疗药物敏感性作用。方法:设计、合成bFGF寡核苷酸,用聚乙烯亚胺(polyemyleneimine,PEI)介导bFGF反义硫代寡核苷酸转染入黑色素瘤B16细胞,MTT法检测bFGF反义硫代寡核苷酸及其与化疗药物联合处理后的细胞增殖率;半定量RT-PCR测定bFGF反义硫代寡核苷酸转染后细胞中bFGF mRNA水平;流式细胞仪分析bFGF反义硫代寡核苷酸诱导的细胞凋亡。结果:bFGF反义硫代寡核苷酸对B16细胞增殖的抑制率为64.8%,且呈剂量依赖效应。B16细胞中bFGF mRNA被bFGF反义硫代寡核苷酸显著降低,为对照细胞的57.9%,且bFGF反义硫代寡核苷酸诱导B16细胞凋亡,凋亡率为41.8%。bFGF反义硫代寡核苷酸转染能显著增强B16细胞对阿霉素、5-氟脲嘧啶及顺铂的敏感性,非特异性硫代寡核苷酸不影响阿霉素、5-氟脲嘧啶及顺铂抑制B16细胞增殖。结论:bFGF反义硫代寡核苷酸显著增强B16细胞的化疗敏感性,表明其可协同化疗药物用于治疗肿瘤。 相似文献
17.
Determining the kinetic constants of arginine uptake by endothelial cells mediated by more than one transporter from linearization
of data as Eadie-Hofstee plots or modeling which does not include the concentration of trace radiolabeled amino acid used
to measure uptake may not be correct. The initial rate of uptake of trace [3H]l-arginine by HUVECs and ECV304 cells in the presence of a range of unlabeled arginine and modifiers was used in nonlinear models to calculate the constants
of arginine uptake using GraphPad Prism. Theoretical plots of uptake derived from constants determined from Eadie-Hofstee
graphs overestimated uptake, whereas those from the nonlinear modeling approach agreed with experimental data. The contribution
of uptake by individual transporters could be modeled and showed that leucine inhibited the individual transporters differently
and not necessarily competitively. N-Ethylmaleimide inhibited only y+ transport, and BCH may be a selective inhibitor of y+L transport. The absence of sodium reduced arginine uptake by y+L transport and reduced the K
m′, whereas reducing sodium decreased arginine uptake by y+ transport without affecting the K
m′. The nonlinear modeling approach using raw data avoided the errors inherent in methods deriving constants from the linearization
of the uptake processes following Michaelian kinetics. This study provides explanations for discrepancies in the literature
and suggests that a nonlinear modeling approach better characterizes the kinetics of amino acid uptake into cells by more
than one transporter. 相似文献
18.
Balkrishen Bhat Normand Heber't Eric G. Marcusson Nicholas M. Dean C. Frank Bennett Muthiah Manoharan 《Nucleosides, nucleotides & nucleic acids》2013,32(6-7):1727-1728
Abstract The synthesis of BODIPY conjugated cationic lipids was achieved in three steps from 3-bromopropane-1,2 diol as the starting material. These compounds were evaluated for their ability to enhance cellular uptake of the antisense oligonucleotides. 相似文献
19.
Metabolic Factors Affecting Enhanced Phosphorus Uptake by Activated Sludge 总被引:2,自引:0,他引:2
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William H. Boughton Richard J. Gottfried Norval A. Sinclair Irving Yall 《Applied microbiology》1971,22(4):571-577
Activated sludges obtained from the Rilling Road plant located at San Antonio, Tex., and from the Hyperion treatment plant located at Los Angeles, Calif., have the ability to remove all of the orthophosphate normally present in Tucson sewage within 3 hr after being added to the waste water. Phosphorus removal was independent of externally supplied sources of energy and ions, since orthophosphate and (32)P radioactivity were readily removed from tap water, glass-distilled water, and deionized water. Phosphorus uptake by Rilling sludge in the laboratory appears to be wholly biological, as it has an optimum pH range (7.7 to 9.7) and an optimum temperature range (24 to 37 C). It was inhibited by HgCl(2), iodoacetic acid, p-chloromercuribenzoic acid, NaN(3), and 2, 4-dinitrophenol (compounds that affect bacterial membrane permeability, sulfhydryl enzymes, and adenosine triphosphate synthesis). Uptake was inhibited by 1% NaCl but was not affected by 10(-3)m ethylenediaminetetraacetic acid disodium salt (a chelating agent for many metallic ions). 相似文献
20.
Masaki Ito Kazumi Hayashi Eru Adachi Tamiko Minamisawa Sadamu Homma Shigeo Koido Kiyotaka Shiba 《PloS one》2014,9(10)
Invocation of cellular immunity by epitopic peptides remains largely dependent on empirically developed protocols, such as interfusion of aluminum salts or emulsification using terpenoids and surfactants. To explore novel vaccine formulation, epitopic peptide motifs were co-programmed with structural motifs to produce artificial antigens using our “motif-programming” approach. As a proof of concept, we used an ovalbumin (OVA) system and prepared an artificial protein library by combinatorially polymerizing MHC class I and II sequences from OVA along with a sequence that tends to form secondary structures. The purified endotoxin-free proteins were then examined for their ability to activate OVA-specific T-cell hybridoma cells after being processed within dendritic cells. One clone, F37A (containing three MHC I and two MHC II OVA epitopes), possessed a greater ability to evoke cellular immunity than the native OVA or the other artificial antigens. The sensitivity profiles of drugs that interfered with the F37A uptake differed from those of the other artificial proteins and OVA, suggesting that alteration of the cross-presentation pathway is responsible for the enhanced immunogenicity. Moreover, F37A, but not an epitopic peptide, invoked cellular immunity when injected together with monophosphoryl lipid A (MPL), and retarded tumor growth in mice. Thus, an artificially synthesized protein antigen induced cellular immunity in vivo in the absence of incomplete Freund''s adjuvant or aluminum salts. The method described here could be potentially used for developing vaccines for such intractable ailments as AIDS, malaria and cancer, ailments in which cellular immunity likely play a crucial role in prevention and treatment. 相似文献