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41.
Pullmannová P Bastos M Bai G Funari SS Lacko I Devínsky F Teixeira J Uhríková D 《Biophysical chemistry》2012,160(1):35-45
Liposome dispersions obtained from the mixture of gemini surfactants of the type alkane-α,ω-diyl-bis(alkyldimethylammonium bromide) and helper lipid DOPC create complexes with DNA showing a regular inner microstructure, identified by small angle X-ray diffraction as condensed lamellar phase (Lαc). In addition to the Lαc phase, a coexisting lamellar phase LB was also identified in the complexes formed, with periodicities in the range ~ 8.8-5.7 nm, at ionic strengths corresponding to 50-200 mM NaCl. The periodicities of LB phase did not correspond to those identified in liposome dispersion without DNA using small angle neutron scattering. The observed phase separation is shown to depend on the interplay between the surface charge density of cationic liposomes, ionic strength and method of complex preparation. The effect of ionic strength on complex formation was studied by isothermal titration calorimetry and zeta potential measurements. High ionic strength reduces the fraction of bound DNA in the complexes, and the isoelectric point is attained at a ratio of DNA/gemini surfactant which is lower than the one that can be estimated by calculation based on nominal charges of CLs and DNA. 相似文献
42.
This work deals with the downstream processing of lipase (EC 3.1.1.3, from Aspergillus niger) using liquid emulsion membrane (LEM) containing reverse micelles for the first time. The membrane phase consisted of surfactants [cetyltrimethylammonium bromide (CTAB) and Span 80] and cosolvents (isooctane and paraffin light oil). The various process parameters for the extraction of lipase from aqueous feed were optimized to maximize activity recovery and purification fold. The mechanism of lipase transport through LEM consisted of three steps namely solubilization of lipase in reverse micelles, transportation of reverse micelles loaded with lipase through the liquid membrane, and release of the lipase into internal aqueous phase. The results showed that the optimum conditions for activity recovery (78.6%) and purification (3.14‐fold) were feed phase ionic strength 0.10 M NaCl and pH 9.0, surfactants concentration (Span 80 0.18 M and CTAB 0.1 M), volume ratio of organic phase to internal aqueous phase 0.9, ratio of membrane emulsion to feed volume 1.0, internal aqueous phase concentration 1.0 M KCl and pH 7.0, stirring speed 450 rpm, and contact time 15 min. This work indicated the feasibility of LEM for the downstream processing of lipase. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012 相似文献
43.
Richard D. Harvey Sandrine Bourgeois Peter Pietzonka Laurent Desire Elias Fattal 《NanoBioTechnology》2005,1(1):71-81
Nanoparticulate complexes of plasmid DNA (pDNA) with cationic liposomes/polymer, of approx 200 nm diameter, were encapsulated
with a high degree of efficiency within calcium pectinate gel beads. Electron microscopy showed the DNA nanocomplexes to be
evenly distributed throughout the gel matrix. Controlled release of pDNA-lipid nanocomplexes was achieved by the action of
pectinase enzymes, whereas release of naked and polymer-complexed DNA was found to be more greatly influenced by the swelling
behavior of the polysaccharide matrices in buffer alone. Physical degradation of pDNA within pectin beads was found to be
accelerated during bead drying, most probably as a result of shear forces generated within the gel matrices by the evaporation
of water. Plasmid complexation with cationic liposomes provided a greater degree of protection for the DNA during bead drying
than complexation with cationic polymer, and was shown to successfully transfect cultured cells after release from the beads,
via the action of pectinase. Observations concerning the physical stability of nanocomplexed pDNA, and its encapsulation within
and release from pectin gel beads, are discussed with reference to the electrostatic interactions existing between the various
components. 相似文献
44.
Piscidin 2 (P2), a 22-residue cationic peptide isolated from the mast cells of hybrid striped bass, has potent antibacterial activities. However, its antifungal properties are not completely understood. In the current study, we investigated the antifungal effects and mode of action of P2. P2 exhibited potent antifungal activity against human pathogenic fungi. To understand the fungicidal properties of P2, we focused on a membrane-active mechanism of the peptide by in vivo and in vitro testing. Flow cytometric analysis using bis-(1,3-dibutylbarbituric acid) trimethine oxonol [DiBAC4(3)] and protoplast regeneration experiments showed that P2 caused fungal membrane damage. Furthermore, fluorescence analysis using 1,6-diphenyl-1,3,5-hexatriene (DPH) revealed that P2 created pores in fungal membranes. These results were confirmed with dye leakage tests by using liposomes composed of phosphatidylcholine/phosphatidylserine (3:1, w/w), which mimicked fungal membranes. The present study indicated that P2 exerts its fungicidal effects by perturbing membrane activities. 相似文献
45.
46.
Tracy A. Stone Gregory B. Cole Huong Q. Nguyen Simon Sharpe Charles M. Deber 《Bioorganic & medicinal chemistry》2018,26(6):1189-1196
Cyclization has been recognized as a valuable technique for increasing the efficacy of small molecule and peptide therapeutics. Here we report the application of a hydrocarbon staple to a rationally-designed cationic antimicrobial peptide (CAP) that acquires increased membrane targeting and interaction vs. its linear counterpart. The previously-described CAP, 6K-F17 (KKKKKK-AAFAAWAAFAA-NH2) was used as the backbone for incorporation of an i to i?+?4 helical hydrocarbon staple through olefin ring closing metathesis. Stapled versions of 6K-F17 showed an increase in non-selective membrane interaction, where the staple itself enhances the degree of membrane interaction and rate of cell death while maintaining high potency against bacterial membranes. However, the higher averaged hydrophobicity imparted by the staple also significantly increases toxicity to mammalian cells. This deleterious effect is countered through stepwise reduction of the stapled 6K-F17’s backbone hydrophobicity through polar amino acid substitutions. Circular dichroism assessment of secondary structure in various bacterial membrane mimetics reveals that a helical structure may improve – but is not an absolute requirement for – antimicrobial activity of 6K-F17. Further, phosphorus-31 static solid state NMR spectra revealed that both non-toxic stapled and linear peptides bind bacterial membranes in a similar manner that does not involve a detergent-like mechanism of lipid removal. The overall results suggest that the technique of hydrocarbon stapling can be readily applied to membrane-interactive CAPs to modulate how they interact and target biological membranes. 相似文献
47.
目的:探讨加味射干麻黄汤对重度支气管哮喘患者血清嗜酸性粒细胞阳离子蛋白(ECP)、脂质过氧化物(LPO)、呼出气一氧化氮(FeNO)及肺功能的影响。方法:选择2015年9月至2017年9月我院接诊的96例重度支气管哮喘患者进行研究,通过随机数表法分为观察组(n=48)和对照组(n=48),在常规治疗基础上,对照组给予异丙托溴铵溶液、甲泼尼龙琥珀酸钠治疗,观察组联合加味射干麻黄汤治疗,均连续治疗4周。比较两组临床疗效、治疗前后中医证候积分、肺功能、血清ECP、LPO及FeNO的变化和不良反应。结果:治疗后,观察组临床疗效总有效率为91.67%(44/48),明显高于对照组的72.92%(35/48)(P0.05);治疗后,两组各中医证候积分、肺功能、血清ECP、LPO及FeNO较治疗前均显著改善(P0.05);中医证候积分中,观察组治疗后喘息、咳嗽、咳痰、胸闷、哮鸣音积分均明显低于对照组,肺功能中,观察组第1秒用力呼出量(FEV1)、FEV1/用力肺活量(FEV1/FVC)均明显高于对照组,呼气峰值流速(PEF)昼夜变异率明显低于对照组,且观察组血清ECP、LPO及FeNO明显比低于对照组,差异均具有统计学意义(P0.05);两组治疗期间不良反应比较无显著差异(P0.05)。结论:在重度支气管哮喘患者中应用加味射干麻黄汤效果显著,可有效改善肺功能及临床症状,其内在机制可能和调节气道慢性炎症反应、氧化应激反应,降低血清ECP、LOP及FeNO的表达相关,且用药安全性高,值得应用推广。 相似文献
48.
Josef Matoušek 《Comparative biochemistry and physiology. Toxicology & pharmacology : CBP》2001,129(3):175-191
The antitumor effect of ribonucleases was studied with animal ribonucleolytic enzymes, bovine pancreatic RNase A, bovine seminal RNase (BS-RNase), onconase and angiogenin. While bovine pancreatic RNase A exerts a minor antitumor effect, BS-RNase and onconase exert significant effects. Angiogenin, as RNase, works in an opposite way, it initiates vascularization of tumors and subsequent tumor growth. Ribonunclease inhibitors are not able to inhibit the antitumor effectiveness of BS-RNase or onconase. However, they do so in the case of pancreatic RNases. Conjugation of BS-RNase with antibodies against tumor antigens (preparation of immunotoxins) like the conjugation of the enzyme with polymers enhances the antitumor activity of the ribonuclease. After conjugation with polymers, the half-life of BS-RNase in blood is extended and its immunogenicity reduced. Recombinant RNases have the same functional activity as the native enzymes. The synthetic genes have also been modified, some of them with gene sequences typical for the BS-RNase parts. Recent experimental efforts are directed to the preparation of ‘humanized antitumor ribonuclease’ that would be structurally similar to human enzyme with minimal immunogenicity and side effects. The angiogenesis of tumors is attempted to be minimized by specific antibodies or anti-angiogenic substances. 相似文献
49.
Muhammad Arba Rahmana E. Kartasasmita 《Journal of biomolecular structure & dynamics》2016,34(2):427-438
A series of cationic porphyrin–anthraquinone hybrids bearing either pyridine, imidazole, or pyrazole rings at the meso-positions have been investigated for their interaction with DNA G-quadruplexes by employing molecular docking and molecular dynamics simulations. Three types of DNA G-quadruplexes were utilized, which comprise parallel, antiparallel, and mixed hybrid topologies. The porphyrin hybrids have a preference to bind with parallel and mixed hybrid structures compared to the antiparallel structure. This preference arises from the end stacking of porphyrin moiety following G-stem and loop binding of anthraquinone tail, which is not found in the antiparallel due to the presence of diagonal and lateral loops that crowd the G-quartet. The binding to the antiparallel, instead, occurred with poorer affinity through both the loop and wide groove. All sites of porphyrin binding were confirmed by 6 ns molecular dynamics simulation, as well as by the negative value of the total binding free energies that were calculated using the MMPBSA method. Free energy analysis shows that the favorable contribution came from the electrostatic term, which supposedly originated from the interaction of either cationic pyridinium, pyrazole, or imidazole groups and the anionic phosphate backbone, and also from the van der Waals energy, which primarily contributed through end stacking interaction. 相似文献
50.