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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Nikolaeva E.D., Goncharov D.B., Shishatskaya E.I.
Заглавие : Effects of H 2O 2-plasma processing on properties of cellular scaffolds made of В«BioplastotanВ» resorbing polyesters
Место публикации : Cellular Transplantation and Tissue Engineering. - 2011. - Vol. 6, Is. 2. - С. 65-69. - ISSN 1815445X (ISSN)
Ключевые слова (''Своб.индексиров.''): adhesion--bioplastotan--cell viability--h 2o 2-plasma processing--resorbing polyesters--scaffold
Аннотация: Produced from В«BioplastotanВ» resorbing polyesters (linear polyesters of hydroxyl derivatives alkanoic acids) scaffolds for cell culturing such as films, pressed 3-D forms and nonwoven fabric from ultrathin fibers are characterized. Two types of polymers - a homopolymer of the 3-hydroxybutyric acid and a copolymer formed by monomers of the 3-hydroxybutyric and 3-hydroxyvalerianic acids are studied. Surface properties of developed polymer scaffolds, sterilized with autoclaving and H 2O 2-plasma processing are compared. It is shown that plasma has beneficial effects resulting in decrease of the watering contact angle and increase of surface hydrophilic properties. Positive effects of H 2O 2-plasma processing of scaffold surface on culturing cell adhesion and viability compared with autoclaving sterilization is demonstrated on NIH 3T3 line fibroblast culturing.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Stepanyuk G.A., Liu Z.J., Markova S.S., Frank L.A., Lee J..., Vysotski E.S., Wang B.C.
Заглавие : Crystal structure of coelenterazine-binding protein from Renilla muelleri at 1.7 angstrom: Why it is not a calcium-regulated photoprotein
Колич.характеристики :6 с
Место публикации : Photochem. Photobiol. Sci.: ROYAL SOC CHEMISTRY, 2008. - Vol. 7, Is. 4. - С. 442-447. - ISSN 1474-905X, DOI 10.1039/b716535h
Примечания : Cited References: 49
Предметные рубрики: HYDROID OBELIA-GENICULATA
AMINO-ACID-SEQUENCE
CA2+-REGULATED PHOTOPROTEINS
RENIFORMIS LUCIFERASE
ENERGY-TRANSFER
CDNA CLONING
BIOLUMINESCENCE
AEQUORIN
PURIFICATION
EXPRESSION
Аннотация: Bioluminescence in the sea pansy Renilla involves two distinct proteins, a Ca2+-triggered coelenterazine-binding protein (CBP), and Renilla luciferase. CBP contains one tightly bound coelenterazine molecule, which becomes available for reaction with luciferase and O-2 only subsequent to Ca2+ binding. CBP belongs to the EF-hand superfamily of Ca2+-binding proteins and contains three "EF-hand" Ca2+-binding sites. The overall spatial structure of recombinant selenomethionine-labeled CBP determined at 1.7 angstrom, is found to approximate the protein scaffold characteristic of the class of Ca2+-regulated photoproteins. Photoproteins however, catalyze molecular oxygen addition to coelenterazine producing a 2-hydroperoxycoelenterazine intermediate, which is stabilized within the binding cavity in the absence of Ca2+. Addition of Ca2+ triggers the bioluminescence reaction. However in CBP this first step of oxygen addition is not allowed. The different amino acid environments and hydrogen bond interactions within the binding cavity are proposed to account for the different properties of the two classes of proteins.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Deng L..., Vysotski E.S., Markova S.V., Liu Z.J., Lee J..., Rose J..., Wang B.C.
Заглавие : All three Ca2+-binding loops of photoproteins bind calcium ions: The crystal structures of calcium-loaded apo-aequorin and apo-obelin
Колич.характеристики :13 с
Место публикации : Protein Sci.: COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT, 2005. - Vol. 14, Is. 3. - С. 663-675. - ISSN 0961-8368, DOI 10.1110/ps.041142905
Примечания : Cited References: 46
Предметные рубрики: RAY CRYSTALLOGRAPHIC ANALYSIS
ANGSTROM RESOLUTION
SEQUENCE-ANALYSIS
CA2+-REGULATED PHOTOPROTEINS
CA2+-DISCHARGED PHOTOPROTEIN
LUMINESCENT PROTEIN
MODULATED PROTEINS
ELECTRON-DENSITY
CLONING
CDNA
Ключевые слова (''Своб.индексиров.''): bioluminescence--ef-hand--fluorescent protein--proton relay--calcium-binding loops--aequorin--obelin--diffraction
Аннотация: The crystal structures of calcium-loaded apo-aequorin and apo-obelin have been determined at resolutions 1.7 Angstrom and 2.2 Angstrom. respectively. A calcium ion is observed in each of the three EF-hand loops that have the canonical calcium-binding sequence, and each is coordinated in the characteristic pentagonal bipyramidal configuration. The calcium-loaded apo-proteins retain the same compact scaffold and overall fold as the unreacted photoproteins containing the bound substrate, 2-hydroperoxycoelenterazine, and also the same as the Ca2+-discharged obelin bound with the product, coelenteramide. Nevertheless, there are easily discerned shifts in both helix and loop regions, and the shifts are not the same between the two proteins. It is suggested that these subtle shifts are the basis of the ability of these photoproteins to sense Ca2+ concentration transients and to produce their bioluminescence response on the millisecond timescale. A mechanism of intrastructural transmission of the calcium signal is proposed.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Stepanyuk G.A., Liu Z.J., Vysotski E.S., Lee J..., Rose J.P., Wang B.C.
Заглавие : Structure based mechanism of the Ca2+ -induced release of coelenterazine from the Renilla binding protein
Колич.характеристики :11 с
Место публикации : Proteins: WILEY-BLACKWELL, 2009. - Vol. 74, Is. 3. - С. 583-593. - ISSN 0887-3585, DOI 10.1002/prot.22173
Примечания : Cited References: 26
Предметные рубрики: GREEN-FLUORESCENT PROTEIN
CRYSTAL-STRUCTURES
RENIFORMIS
LUCIFERASE
BIOLUMINESCENCE
PURIFICATION
ANGSTROM
MUELLERI
Ключевые слова (''Своб.индексиров.''): bioluminescence--ef-hand--coelenteramider--luciferase--ca2+-binding protein
Аннотация: The crystal structure of the Ca2+-loaded coelenterazine binding protein from Renilla muelleri in its apo-state has been determined at resolution 1.8 angstrom. Although calcium binding hardly affects the compact scaffold and overall fold of the structure before calcium addition, there are easily discerned shifts in the residues that were interacting with the coelenterazine and a repositioning of helices, to expose a cavity to the external solvent. Altogether these changes offer a straightforward explanation for how following the addition of Ca2+, the coelenterazine could escape and become available for bioluminescence on Renilla luciferase. A docking computation supports the possibility of a luciferase-binding protein complex.
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Вид документа :
Шифр издания :
Автор(ы) : Volova T. G., Goncharov D. B., Nikolaeva E. D., Shishatskaya E. I.
Заглавие : Electrospinning of degradable phas: Process, properties, applications
Место публикации :: Nova Science Publishers, Inc., 2017. - С. 1-56
Ключевые слова (''Своб.индексиров.''): biological and physical/mechanical properties--cell proliferation--electrospinning--nonwoven membranes--pha--scaffolds--skin regeneration--ultrafine fibers--wound dressings
Аннотация: An integrated study has been performed to investigate the process of formation of ultrafine fibers and nonwoven membranes by electrospinning from natural degradable polymers-polyhydroxyalkanoates (PHAs); physical, mechanical, and biological properties of the products have been studied. Then, electrospinning was used to prepare ultrafine fibers from PHAs with different compositions: P(3HB) and its copolymers P(3HB-co-4HB), P(3HB-co-3HV), and P(3HB-co-3HHx). The main process parameters, that influence UF-fiber diameter and properties of fibrous non-woven membranes) (polymer concentration, solution feeding rate, working distance, and applied voltage), were investigated and their effects evaluated. This study was the first to compare biological and physical/mechanical parameters of PHAs with different chemical compositions as dependent upon the fractions of monomers, constituting the polymers and fiber orientation. Electrospun polymer membranes, prepared from the [P(3HB-co-4HB)], were tested as wound dressings. The developed nonwoven membranes can be used as the equivalent of collagen skine dressings in the treatment of burns of degree II. Experiments on laboratory animals with model skin defects showed, that the membranes fitted the wound shape good, protected the wound from external influences, and facilitated wound healing, promoting fast repair. The successful experiments on laboratory animals were followed by pilot clinical trials of nanomembranes, comprised of PHA membranes as wound dressings in the treatment of septic wounds. During the regeneration phase, PHA membranes served as a scaffold for the new tissue on the skin and filled out soft tissue defects. The formation of the uniform and sufficiently vascularized tissue is a prerequisite for quicker wound healing and can serve as a basis for the subsequent skin grafting and spontaneous re-epithelialization of superficial wounds. The wound dressing, tested in this clinical trial, performs important physiological functions of natural skin, provides a barrier against secondary infection, reduces fluid loss, and, at the same time, does not keep the air out. © 2017 Nova Science Publishers, Inc.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Lukyanenko K. A., Denisov I. A., Yakimov A. S., Esimbekova E. N., Belousov K. I., Bukatin A. S., Kukhtevich I. V., Sorokin V. V., Evstrapov A. A., Belobrov P. I.
Заглавие : Analytical Enzymatic Reactions in Microfluidic Chips
Колич.характеристики :6 с
Коллективы : Russian Science Foundation [15-19-10041]
Место публикации : Appl. Biochem. Microbiol.: MAIK NAUKA/INTERPERIODICA/SPRINGER, 2017. - Vol. 53, Is. 7. - С. 775-780. - ISSN 0003-6838, DOI 10.1134/S0003683817070043. - ISSN 1573-8183(eISSN)
Примечания : Cited References:15. - The study was supported by a grant from the Russian Science Foundation (project No. 15-19-10041).
Предметные рубрики: BIOAVAILABLE HEAVY-METALS
DEVICES
POINT
LAB
Ключевые слова (''Своб.индексиров.''): bioluminescence--luciferase--microfluidics--microfluidic chip--enzymatic--bioassay
Аннотация: A number of approaches have been proposed and tested to transfer enzymatic reactions into the functional elements of microfluidic chips on the example of the bienzyme bioluminescent reaction involving NAD(P)H:FMN-oxidoreductase and luciferase. Measurement of the catalytic activity of these enzymes (under the influence of pollutants) is the basis of enzymatic bioassay of various liquids. It was found that all of the components of the reaction must be placed in the same cell of the chip to improve the reproducibility of the measurements. The use of starch gel as a carrier for immobilization and gelatin as a scaffold in the reactor of the chip enables the preservation of enzyme activity in the course of sealing the chip at room temperature. It is shown that the components of the reaction should be vigorously stirred in a microfluidic chip reactor to improve the efficiency of the analysis. As a result of the studies, a prototype of microfluidic chip based on the enzymatic bioluminescent reaction is proposed. It is characterized by a detection limit of copper sulfate of 3 mu M that corresponds to the sensitivity of traditional lux-biosensors based on living cells. The analysis time is reduced to 1 min, and the analysis can be performed by individuals without special laboratory skills.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Denisov I., Lukyanenko K., Yakimov A., Kukhtevich I., Esimbekova E., Belobrov P.
Заглавие : Disposable luciferase-based microfluidic chip for rapid assay of water pollution
Место публикации : Lumin. - 2018. - Vol. 33, Is. 6. - С. 1054-1061. - ISSN 15227235 (ISSN) , DOI 10.1002/bio.3508
Ключевые слова (''Своб.индексиров.''): bioassay--lab-on-a-chip--luciferase--microfluidics--solvent bonding
Аннотация: In the present study, we demonstrate the use of a disposable luciferase-based microfluidic bioassay chip for environmental monitoring and methods for fabrication. The designed microfluidic system includes a chamber with immobilized enzymes of bioluminescent bacteria Photobacterium leiognathi and Vibrio fischeri and their substrates, which dissolve after the introduction of the water sample and thus activate bioluminescent reactions. Limits of detection for copper (II) sulfate, 1,3-dihydroxybenzene and 1,4-benzoquinone for the proposed microfluidic biosensor measured 3 ?M, 15 mM, and 2 ?M respectively, and these values are higher or close to the level of conventional environmental biosensors based on lyophilized bacteria. Approaches for entrapment of enzymes on poly(methyl methacrylate) (PMMA) plates using a gelatin scaffold and solvent bonding of PMMA chip plates under room temperature were suggested. The proposed microfluidic system may be used with some available luminometers and future portable luminescence readers. © 2018 John Wiley & Sons, Ltd.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Burakova L. P., Vysotski E. S.
Заглавие : Recombinant Ca2+-regulated photoproteins of ctenophores: current knowledge and application prospects
Место публикации : Appl. Microbiol. Biotechnol.: Springer Verlag, 2019. - Vol. 103, Is. 15. - С. 5929-5946. - ISSN 01757598 (ISSN) , DOI 10.1007/s00253-019-09939-0
Аннотация: Bright bioluminescence of ctenophores is conditioned by Ca2+-regulated photoproteins. Although they share many properties characteristic of hydromedusan Ca2+-regulated photoproteins responsible for light emission of marine animals belonging to phylum Cnidaria, a substantial distinction still exists. The ctenophore photoproteins appeared to be extremely sensitive to light—they lose the ability for bioluminescence on exposure to light over the entire absorption spectrum. Inactivation is irreversible because keeping the inactivated photoprotein in the dark does not recover its activity. The capability to emit light can be restored only by incubation of inactivated photoprotein with coelenterazine in the dark at alkaline pH in the presence of oxygen. Although these photoproteins were discovered many years ago, only the cloning of cDNAs encoding these unique bioluminescent proteins in the early 2000s has provided a new impetus for their studies. To date, cDNAs encoding Ca2+-regulated photoproteins from four different species of luminous ctenophores have been cloned. The amino acid sequences of ctenophore photoproteins turned out to completely differ from those of hydromedusan photoproteins (identity less than 29%) though also similar to them having three EF-hand Ca2+-binding sites. At the same time, these photoproteins reveal the same two-domain scaffold characteristic of hydromedusan photoproteins. This review is an attempt to systemize and critically evaluate the data scattered through various articles regarding the structural features of recombinant light-sensitive Ca2+-regulated photoproteins of ctenophores and their bioluminescent and physicochemical properties as well as to compare them with those of hydromedusan photoproteins. In addition, we also discuss the prospects of their biotechnology applications. © 2019, Springer-Verlag GmbH Germany, part of Springer Nature.
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Вид документа : Статья из журнала
Шифр издания :
Автор(ы) : Shishatskaya E., Nemtsev I., Lukyanenko A., Vasiliev A., Kiselev E., Sukovatyi A., Volova T.
Заглавие : Polymer Films of Poly-3-hydroxybutyrate Synthesized by Cupriavidus necator from Different Carbon Sources
Место публикации : J. Polym. Environ.: Springer, 2021. - Vol. 29, Is. 3. - С. 837-850. - ISSN 15662543 (ISSN), DOI 10.1007/s10924-020-01924-3
Аннотация: Films were prepared from 2% solutions of biodegradable poly-3-hydroxybutyrate [P(3HB)] and investigated. The polymer was synthesized by the Cupriavidus necator B-10646 bacterium cultivated using various carbon sources (glucose and glycerol of different degrees of purity, containing 0.3 to 17.93% impurities). Glycerol as the substrate influenced molecular-weight properties and crystallinity of the polymer without affecting its temperature characteristics. The P(3HB) specimens synthesized from glycerol had reduced Mw (300–400 kDa) and degree of crystallinity (50–55%) compared to the specimens synthesized from glucose (860 kDa and 76%, respectively). The low-crystallinity P(3HB) specimens, regardless of the degree of purity of glycerol, produced a beneficial effect on the properties of polymer films, which had a better developed folded surface and increased hydrophilicity. The values of the highest roughness (Ra) of the films synthesized from glycerol were 1.8 to 4.0 times lower and the water angles 1.4–1.6 times smaller compared to the films synthesized from glucose (71.75 nm and 87.4°, respectively). Those films performed better as cell scaffolds: the number of viable NIH fibroblasts was 1.7–1.9 times higher than on polystyrene (control) or films of P(3HB) synthesized from glucose. © 2020, Springer Science+Business Media, LLC, part of Springer Nature.
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