Влияние стабилизирующих добавок на свойства коллоидных частиц альгината кальция

In the process of 210-240 nm calcium alginate nanoparticles (AlgNP) non-emulsion synthesis from solutions with the 50-250-fold weight excess of stabilizers, polyethelyneglycol (PEG 1500) and Tween 80, as compared to polysaccharide, their molecules are incorporated in the forming gel in minimal concenrations (less than 64 ± 14 and 19 ± 9 mg/g on dry weight basis, accordingly). Their presence in solution decreases AlgNP size and stabilizes colloids. PEG 1500 macromolecules embedded into AlgNP slightly decrease gel polarity, while in AlgNP obtained with Tween 80, the local pockets formed by surfactant hydrophobic tails have been found.В процессе неэмульсионного синтеза частиц альгината кальция (АлгНЧ) диаметром 210-240 нм из растворов с 50-250-кратным избытком стабилизаторов, полиэтиленгликоля (ПЭГ 1500) и Твин 80, по отношению к массе полисахарида их молекулы включаются в гель в минимальных концентрациях (не более 64 ± 14 и 19 ± 9 мг/г в пересчете на сухую массу соответственно). Однако их присутствие в растворе уменьшает размер АлгНЧ и стабилизирует образующийся коллоид. Включенные в АлгНЧ макромолекулы ПЭГ 1500 незначительно уменьшают полярность геля, а в АлгНЧ, полученных с Твин 80, присутствуют локальные области, сформированные гидрофобными хвостами сурфактанта

ВЛИЯНИЕ СТРУКТУРЫ СОПОЛИМЕРА И МУЛЬТИСЛОЙНОЙ ОБОЛОЧКИ НА АГРЕГАТИВНУЮ УСТОЙЧИВОСТЬ НАНОКАПСУЛ (ХИТОЗАН-прив-ПОЛИЭТИЛЕНГЛИКОЛЬ / ПОЛИСТИРОЛСУЛЬФОНАТ)n

Nanocapsules consisted of magnetic core and polyelectrolyte shell on the basis of chitosan or its polyethylene glycol (PEG) grafted copolymers (CH) and polystyrene sulfonate (PSS) were obtained by layer-by-layer assembly. The influence of the number of CH/PSS bilayers in a shell, polymerization degree of CH, and the presence of grafted PEG chains on aggregative and sedimentation stability of the obtained nanocapsules in different media (water, 0.9 % NaCl solution, fetal bovine serum (FBS)) was discussed. It was shown that the diameter of PSS-terminated (CH/PSS)n nanocapsules in FBS does not change at 37 °С for 6–8 h. Adsorption of bovine serum albumin on the surface of the layer-by-layer nanocapsules stabilizes them in isotonic 0.9 % solution of NaCl and FBS. Co-introduction of copolymer and albumin in the shells improves generally the aggregative stability of nanocapsules in water and allows one to obtain stable nanocapsules with magnetic core for fewer adsorption cycles.Нанокапсулы, состоящие из магнитного ядра и мультислойной полиэлектролитной оболочки на основе хитозана и его сополимеров с полиэтиленгликолем (ХН) и полистиролсульфоната (ПСС), получены методом послойной сборки. Изучено влияние числа бислоев ХН/ПСС в оболочке, степени полимеризации хитозана и наличия боковых цепей полиэтиленгликоля на агрегативно-седиментационную устойчивость нанокапсул в различных средах (воде, 0,9 %-ном растворе NaCl, эмбриональной телячьей сыворотке (ЭТС)). Показано, что диаметр нанокапсул (ХН/ПСС)n со слоем ПСС на поверхности в ЭТС не изменяется при 37 °С в течение 6–8 ч. Адсорбция альбумина на поверхности нанокапсул позволяет стабилизировать их в изотоническом 0,9 %-ном растворе NaCl и ЭТС. Совместное использование сополимера и альбумина в составе оболочек в целом улучшает агрегативно-седиментационную устойчивость нанокапсул в воде и позволяет получить устойчивые нанокапсулы с магнитным ядром за меньшее число циклов адсорбции

Electrochemistry of nanozeolite-immobilized cytochrome c in aqueous and nonaqueous solutions

peer-reviewedThe electrochemical properties of cytochrome c (cyt c) immobilized on multilayer nanozeolite-modified electrodes have been examined in aqueous and nonaqueous solutions. Layers of Linde type-L zeolites were assembled on indium tin oxide (ITO) glass electrodes followed by the adsorption of cyt c, primarily via electrostatic interactions, onto modified ITO electrodes. The heme protein displayed a quasi-reversible response in aqueous solution with a redox potential of +324 mV (vs NHE), and the surface coverage (Gamma*) increased linearly for the first four layers and then gave a nearly constant value of 200 pmol cm(-2). On immersion of the modified electrodes in 95% (v/v) nonaqueous solutions, the redox potential decreased significantly, a decrease that originated from changes in both the enthalpy and entropy of reduction. On reimmersion of the modified electrode in buffer, the faradic response immediately returned to its original value. These results demonstrate that nanozeolites are potential stable supports for redox proteins and enzymes.ACCEPTEDpeer-reviewe

СИНТЕЗ И ФУНКЦИОНАЛИЗАЦИЯ ПОВЕРХНОСТИ МАГНИТНЫХ НАНОЧАСТИЦ (Mg, Zn)xFe3–xO4

The nanoparticles of (Mg, Zn)xFe3–xO4 (x ≤ 0.3) solid solutions have been prepared by coprecipitation with Na2CO3 from solutions of salts. For a number of compositions (Mg0,1Fe2,9O4, Mg0,05Zn0,1Fe2,85O4, Zn0,18Fe2,82O4), an increase of saturation magnetization has been detected, as compared to non-substituted magnetite (MS = 64 emu/g). This can be explained by the tendency of zinc and small amounts of magnesium ions to occupy preferentially tetrahedral sites of the magnetite lattice. In the case of zinc and magnesium joint substitution in the (Mg, Zn)xFe3–xO4 system up to x = 0.3, the values of saturation magnetization decrease slightly comparing to that of magnetite, but remain constant (MS ≈ 58 emu/g). By ultrasound assisted dispersion of nanopowders into polyelectrolyte aqueous solutions, colloidal solutions of non-agglomerated nanoparticles have been prepared. Тhe nanoparticles modified with a layer of positively charged polyelectrolyte demonstrate the best sedimentation stability up to 45 days. Their hydrodynamic diameter is lower than 200 nm, with predominance of the fraction with the size of 40–80 nm.Методом соосаждения с Na2CO3 из растворов солей получены наночастицы твердых растворов в системе (Mg, Zn)xFe3–xO4 (x ≤ 0,3). Для ряда составов (Mg0,1Fe2,9O4, Mg0,05Zn0,1Fe2,85O4, Zn0,18Fe2,82O4) обнаружен рост намагниченности насыщения по сравнению с незамещенным магнетитом (MS = 64 А·м2 ·кг–1), что объясняется склонностью ионов цинка и малых количеств ионов магния занимать в решетке магнетита преимущественно тетраэдрические пустоты. В случае совместного замещения цинком и магнием в системе (Mg, Zn)xFe3–xO4 вплоть до х = 0,3 значения намагниченности насыщения незначительно снижаются относительно магнетита, однако остаются на постоянном уровне (MS ≈ 58 А·м2 ·кг–1), предположительно благодаря стабилизирующему влиянию ионов магния. Ультразвуковым диспергированием нанопорошков в водных растворах полиэлектролитов получали коллоидные растворы наночастиц в неагломерированном состоянии. Наилучшей седиментационной устойчивостью (45 дней) обладают наночастицы, модифицированных слоем положительно заряженного полиэлектролита. Их гидродинамический диаметр не превышает 200 нм, причем преобладает фракция частиц с размерами 40–80 нм.

Layer-by-Layer Assembled Antisense DNA Microsponge Particles for Efficient Delivery of Cancer Therapeutics

Antisense oligonucleotides can be employed as a potential approach to effectively treat cancer. However, the inherent instability and inefficient systemic delivery methods for antisense therapeutics remain major challenges to their clinical application. Here, we present a polymerized oligonucleotides (ODNs) that self-assemble during their formation through an enzymatic elongation method (rolling circle replication) to generate a composite nucleic acid/magnesium pyrophosphate sponge-like microstructure, or DNA microsponge, yielding high molecular weight nucleic acid product. In addition, this densely packed ODN microsponge structure can be further condensed to generate polyelectrolyte complexes with a favorable size for cellular uptake by displacing magnesium pyrophosphate crystals from the microsponge structure. Additional layers are applied to generate a blood-stable and multifunctional nanoparticle via the layer-by-layer (LbL) assembly technique. By taking advantage of DNA nanotechnology and LbL assembly, functionalized DNA nanostructures were utilized to provide extremely high numbers of repeated ODN copies for efficient antisense therapy. Moreover, we show that this formulation significantly improves nucleic acid drug/carrier stability during in vivo biodistribution. These polymeric ODN systems can be designed to serve as a potent means of delivering stable and large quantities of ODN therapeutics systemically for cancer treatment to tumor cells at significantly lower toxicity than traditional synthetic vectors, thus enabling a therapeutic window suitable for clinical translation.United States. Dept. of Defense. Ovarian Cancer Research Program (Teal Innovator Award Grant OC120504)Natural Sciences and Engineering Research Council of Canada (Postdoctoral Fellowship)National Institutes of Health (U.S.) (Ruth L. Kirschstein National Research Service Award 1F32EB017614-01)National Science Foundation (U.S.). Graduate Research Fellowshi

Enzymatic Analysis of Recombinant Japanese Encephalitis Virus NS2B(H)-NS3pro Protease with Fluorogenic Model Peptide Substrates

Background Japanese encephalitis virus (JEV), a member of the Flaviviridae family, causes around 68,000 encephalitis cases annually, of which 20–30% are fatal, while 30–50% of the recovered cases develop severe neurological sequelae. Specific antivirals for JEV would be of great importance, particularly in those cases where the infection has become persistent. Being indispensable for flaviviral replication, the NS2B-NS3 protease is a promising target for design of anti-flaviviral inhibitors. Contrary to related flaviviral proteases, the JEV NS2B-NS3 protease is structurally and mechanistically much less characterized. Here we aimed at establishing a straightforward procedure for cloning, expression, purification and biochemical characterization of JEV NS2B(H)-NS3pro protease. Methodology/Principal Findings The full-length sequence of JEV NS2B-NS3 genotype III strain JaOArS 982 was obtained as a synthetic gene. The sequence of NS2B(H)-NS3pro was generated by splicing by overlap extension PCR (SOE-PCR) and cloned into the pTrcHisA vector. Hexahistidine-tagged NS2B(H)-NS3pro, expressed in E. coli as soluble protein, was purified to >95% purity by a single-step immobilized metal affinity chromatography. SDS-PAGE and immunoblotting of the purified enzyme demonstrated NS2B(H)-NS3pro precursor and its autocleavage products, NS3pro and NS2B(H), as 36, 21, and 10 kDa bands, respectively. Kinetic parameters, Km and kcat, for fluorogenic protease model substrates, Boc-GRR-amc, Boc-LRR-amc, Ac-nKRR-amc, Bz-nKRR-amc, Pyr-RTKR-amc and Abz-(R)4SAG-nY-amide, were obtained using inner filter effect correction. The highest catalytic efficiency kcat/Km was found for Pyr-RTKR-amc (kcat/Km: 1962.96±85.0 M−1 s−1) and the lowest for Boc-LRR-amc (kcat/Km: 3.74±0.3 M−1 s−1). JEV NS3pro is inhibited by aprotinin but to a lesser extent than DEN and WNV NS3pro. Conclusions/Significance A simplified procedure for the cloning, overexpression and purification of the NS2B(H)-NS3pro was established which is generally applicable to other flaviviral proteases. Kinetic parameters obtained for a number of model substrates and inhibitors, are useful for the characterization of substrate specificity and eventually for the design of high-throughput assays aimed at antiviral inhibitor discovery

Properties of layer-by-layer coatings based on poly(allylamine)-graft-poly(ethylene glycol) copolymers and dextran sulfate

Chemistry of molecular sуstem