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Production of purified polyhydroxyalkanoates (PHAs) for applications in contact with blood / V. I. Sevastianov [et al.] // Journal of Biomaterials Science, Polymer Edition. - 2003. - Vol. 14, Is. 10. - P1029-1042, DOI 10.1163/156856203769231547 . - ISSN 0920-5063
Кл.слова (ненормированные):
?-hydroxy acids -- Endotoxins -- Hemocompatibility -- Poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) -- Polyhydroxyalkanoates (PHAs) -- Polyhydroxybutyrate (PHB) -- bacterium lipopolysaccharide -- carbon -- complement -- copolymer -- hydroxyacid -- long chain fatty acid -- poly(3 hydroxybutyric acid) -- polyhydroxyalkanoic acid -- valeric acid derivative -- adult -- article -- biofilm -- biotechnology -- blood analysis -- blood clotting -- blood compatibility -- cell function -- chemical analysis -- chemical composition -- complement activation -- concentration (parameters) -- controlled study -- gas chromatography -- hemostasis -- human -- human cell -- mass spectrometry -- micromorphology -- nonhuman -- priority journal -- purification -- quantitative analysis -- sampling -- synthesis -- thrombocyte adhesion -- Wautersia eutropha -- Biocompatible Materials -- Blood -- Blood Coagulation Tests -- Chromatography, Gas -- Complement Activation -- Cupriavidus necator -- Fatty Acids -- Humans -- Platelet Adhesiveness -- Polyesters -- Surface Properties
Аннотация: Samples of olyhydroxyalkanoates (PHAs), polyhydroxybutyrate (PHB) and copolymers poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) with 4 and 18 mol% hydroxyvalerate, synthesized by the bacteria Ralstonia eutropha B5786, were investigated. PHA films in contact with blood did not activate the hemostasis system at the level of cell response, but they did activate the coagulation system and the complement reaction. To detect biologically-active components in the PHAs, a detailed analysis of the composition of the polymers was conducted. Gas chromatography-mass spectrometry revealed long-chain fatty acids (FAs) in the tested PHAs. Their total concentration in the polymer ranged from tenths of mol% to 2-3 mol%, depending on the purification method. C16:0 constituted the largest proportion, up to 70%. Of the long-chain hydroxy acids, only ?-OH-C14:0 was detected and it did not exceed 0.06 mol%. The analysis of the hemocompatibility properties of the PHAs purified by a specialized procedure, including the quantitative and morphological estimation of platelets adherent to the surface of polymer films, the plasma recalcification time and complement activation studies, indicated that PHB and PHBV can be used in contact with blood. It has been found out that the lipopolysaccharides of bacteria producing PHAs, which contain mostly long-chain hydroxy acids, can be the factor activating the hemostasis systems. Thus, the technology of PHA purification must satisfy rather stringent specific requirements.

Scopus
Держатели документа:
Inst. of Transplantol. Artif. Organs, Russian Ministry of Health, Shchukinskaya 1, 123182 Moscow, Russian Federation
Inst. of Biophys. of Siberian Branch, Russian Academy of Sciences, Akademgorodok, Krasnoyarsk 660036, Russian Federation : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Sevastianov, V.I.; Perova, N.V.; Shishatskaya, E.I.; Kalacheva, G.S.; Volova, T.G.

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Production of purified polyhydroxyalkanoates (PHAs) for applications in contact with blood [Text] / V. I. Sevastianov [et al.] // J. Biomater. Sci.-Polym. Ed. - 2003. - Vol. 14, Is. 10. - P. 1029-1042, DOI 10.1163/156856203769231547. - Cited References: 34 . - ISSN 0920-5063

РУБ Engineering, Biomedical + Materials Science, Biomaterials + Polymer Science
Рубрики:
BIODEGRADABLE POLYESTERS
POLYMERS
Кл.слова (ненормированные):
polyhydroxyalkanoates (PHAs) -- polyhydroxybutyrate (PHB) -- poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) -- endotoxins -- beta-hydroxy acids -- hemocompatibility
Аннотация: Samples of olyhydroxyalkanoates (PHAs), polyhydroxybutyrate (PHB) and copolymers poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) with 4 and 18 mol% hydroxyvalerate, synthesized by the bacteria Ralstonia eutropha B5786, were investigated. PHA films in contact with blood did not activate the hemostasis system at the level of cell response, but they did activate the coagulation system and the complement reaction. To detect biologically-active components in the PHAs, a detailed analysis of the composition of the polymers was conducted. Gas chromatography-mass spectrometry revealed long-chain fatty acids (FAs) in the tested PHAs. Their total concentration in the polymer ranged from tenths of mol% to 2-3 mol%, depending on the purification method. C-16:0 constituted the largest proportion, up to 70%. Of the long-chain hydroxy acids, only beta-OH-C-14:0 was detected and it did not exceed 0.06 mol%. The analysis of the hemocompatibility properties of the PHAs purified by a specialized procedure, including the quantitative and morphological estimation of platelets adherent to the surface of polymer films, the plasma recalcification time and complement activation studies, indicated that PHB and PHBV can be used in contact with blood. It has been found out that the lipopolysaccharides of bacteria producing PHAs, which contain mostly long-chain hydroxy acids, can be the factor activating the hemostasis systems. Thus, the technology of PHA purification must satisfy rather stringent specific requirements.

WOS
Держатели документа:
Russian Acad Sci, Inst Biophys, Siberian Branch, Krasnoyarsk 660036, Russia
Russian Minist Hlth, Inst Transplantol Artificial Organs, Moscow 123182, Russia
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Sevastianov, V.I.; Perova, N.V.; Shishatskaya, E.I.; Kalacheva, G.S.; Volova, T.G.

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Simultaneous Genotyping of Four Single Nucleotide Polymorphisms Associated with Risk Factors of Hemostasis Disorders [Text] / E. E. Bashmakova, V. V. Krasitskaya, L. A. Frank // Comb. Chem. High Throughput Screen. - 2015. - Vol. 18, Is. 10. - P930-936, DOI 10.2174/1386207318666150917095903. - Cited References:20. - The study was supported by the grant 14-14-01119 of the Russian Science Foundation. . - ISSN 1386-2073. - ISSN 1875-5402

РУБ Biochemical Research Methods + Chemistry, Applied + Pharmacology &
Рубрики:
ALLELE-SPECIFIC PCR
FACTOR-V-LEIDEN
BIOLUMINESCENT IMMUNOASSAY
Кл.слова (ненормированные):
SNP detection -- PEXT reaction -- photoprotein obelin -- bioluminescent -- microassay -- multiplex PCR
Аннотация: Multiplex simultaneous genotyping technique was developed for four polymorphisms in genes coding for blood coagulation factors and homocysteine metabolism which are considered as thrombophilia related mutations: FV Leiden, FII G20210A, MTHFR C677T, and FVII G10976A. It is based on primer extension reaction with the following bioluminescent solid-phase microassay. At that, two different in bioluminescence obelin mutants were applied to simultaneous detection of two gene allelic variants. The assay is carried out in microtiter plate format and provides fast and reliable genotyping of four single nucleotide polymorphisms in four different genes within 2.5 hours. A large number of clinical samples were analyzed and the obtained results were found to be in complete correlation with those obtained by using conventional RT-PCR techniques.

WOS
Держатели документа:
Russian Acad Sci, Siberian Branch, Inst Biophys, Photobiol Lab, Krasnoyarsk 660036, Russia.
Siberian Fed Univ, Krasnoyarsk 660041, Russia.

Доп.точки доступа:
Bashmakova, Eugenia E.; Krasitskaya, Vasilisa V.; Frank, Ludmila A.; Russian Science Foundation [14-14-01119]

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Simultaneous genotyping of four single nucleotide polymorphisms associated with risk factors of hemostasis disorders / E. E. Bashmakova, V. V. Krasitskaya, L. A. Frank // Comb. Chem. High Throughput Screen. - 2015. - Vol. 18, Is. 10. - P930-936 . - ISSN 1386-2073
Кл.слова (ненормированные):
Bioluminescent microassay -- Multiplex PCR -- PEXT reaction -- Photoprotein obelin -- SNP detection
Аннотация: Multiplex simultaneous genotyping technique was developed for four polymorphisms in genes coding for blood coagulation factors and homocysteine metabolism which are considered as thrombophilia related mutations: FV Leiden, FII G20210A, MTHFR C677T, and FVII G10976A. It is based on primer extension reaction with the following bioluminescent solid-phase microassay. At that, two different in bioluminescence obelin mutants were applied to simultaneous detection of two gene allelic variants. The assay is carried out in microtiter plate format and provides fast and reliable genotyping of four single nucleotide polymorphisms in four different genes within 2.5 hours. A large number of clinical samples were analyzed and the obtained results were found to be in complete correlation with those obtained by using conventional RT-PCR techniques. © 2015 Bentham Science Publishers.

Scopus
Держатели документа:
Photobiology Laboratory, Institute of Biophysics, Russian Academy of Sciences, Siberian Branch, Krasnoyarsk, Russian Federation
Siberian Federal University, Svobodnii Ave., 79, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Bashmakova, E. E.; Krasitskaya, V. V.; Frank, L. A.

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