Expression in escherichia coli and purification of folded rDer p 20, the arginine kinase from dermatophagoides pteronyssinus: A possible biomarker for allergic asthma
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01.01.2021 |
Sarzsinszky E.
Lupinek C.
Vrtala S.
Huang H.J.
Hofer G.
Keller W.
Chen K.W.
Panaitescu C.B.
Resch-Marat Y.
Zieglmayer P.
Zieglmayer R.
Lemell P.
Horak F.
Duchêne M.
Valenta R.
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Allergy, Asthma and Immunology Research |
10.4168/AAIR.2021.13.1.154 |
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Copyright © 2021 The Korean Academy of Asthma, Allergy and Clinical Immunology. Arginine kinase (AK) was first identified as an allergen in the Indian-meal moth and subsequently shown to occur as allergen in various invertebrates and shellfish. The cDNA coding for AK from the house dust mite (HDM) species Dermatophagoides pteronyssinus, Der p 20, has been isolated, but no recombinant Der p 20 (rDer p 20) allergen has been produced and characterized so far. We report the expression of Der p 20 as recombinant protein in Escherichia coli. rDer p 20 was purified and shown to be a monomeric, folded protein by size exclusion chromatography and circular dichroism spectroscopy, respectively. Using AK-specific antibodies, Der p 20 was found to occur mainly in HDM bodies, but not in fecal particles. Thirty percent of clinically well-characterized HDM allergic patients (n = 98) whose immunoglobulin E (IgE) reactivity profiles had been determined with an extensive panel of purified HDM allergens (Der f 1, 2; Der p 1, 2, 4, 5, 7, 10, 11, 14, 15, 18, 21, 23 and 37) showed IgE reactivity to Der p 20. IgE reactivity to Der p 20 was more frequently associated with lung symptoms. AKs were detected in several invertebrates with specific antibodies and Der p 20 showed IgE cross-reactivity with AK from shrimp (Litopenaeus vannamei). Thus, Der p 20 is a cross-reactive HDM allergen and may serve as a diagnostic marker for HDM-induced lung symptoms such as asthma.
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Expression in escherichia coli and purification of folded rDer p 20, the arginine kinase from dermatophagoides pteronyssinus: A possible biomarker for allergic asthma
|
01.01.2021 |
Sarzsinszky E.
Lupinek C.
Vrtala S.
Huang H.J.
Hofer G.
Keller W.
Chen K.W.
Panaitescu C.B.
Resch-Marat Y.
Zieglmayer P.
Zieglmayer R.
Lemell P.
Horak F.
Duchêne M.
Valenta R.
|
Allergy, Asthma and Immunology Research |
10.4168/AAIR.2021.13.1.154 |
0 |
Ссылка
Copyright © 2021 The Korean Academy of Asthma, Allergy and Clinical Immunology. Arginine kinase (AK) was first identified as an allergen in the Indian-meal moth and subsequently shown to occur as allergen in various invertebrates and shellfish. The cDNA coding for AK from the house dust mite (HDM) species Dermatophagoides pteronyssinus, Der p 20, has been isolated, but no recombinant Der p 20 (rDer p 20) allergen has been produced and characterized so far. We report the expression of Der p 20 as recombinant protein in Escherichia coli. rDer p 20 was purified and shown to be a monomeric, folded protein by size exclusion chromatography and circular dichroism spectroscopy, respectively. Using AK-specific antibodies, Der p 20 was found to occur mainly in HDM bodies, but not in fecal particles. Thirty percent of clinically well-characterized HDM allergic patients (n = 98) whose immunoglobulin E (IgE) reactivity profiles had been determined with an extensive panel of purified HDM allergens (Der f 1, 2; Der p 1, 2, 4, 5, 7, 10, 11, 14, 15, 18, 21, 23 and 37) showed IgE reactivity to Der p 20. IgE reactivity to Der p 20 was more frequently associated with lung symptoms. AKs were detected in several invertebrates with specific antibodies and Der p 20 showed IgE cross-reactivity with AK from shrimp (Litopenaeus vannamei). Thus, Der p 20 is a cross-reactive HDM allergen and may serve as a diagnostic marker for HDM-induced lung symptoms such as asthma.
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The frequency and species composition of vaginal bacterial carriage in the third trimester of gestation
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01.01.2018 |
Naumenko N.
Мorozova О.
Kuksyuk P.
Lyakhova О.
Aleksandrov L.
Аstsaturova О.
Belova А.
Nikonov A.
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Voprosy Ginekologii, Akusherstva i Perinatologii |
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© 2018, Dynasty Publishing House. All rights reserved. The objective. To specify the frequency and species composition of vaginal bacterial carriage in pregnant women at the term of gestation 35–37 wks. Patients and methods. We examined 800 pregnant women, who were followed-up on an outpatient basis at terms of gestation 35–37 wks. Cultural examination of the content of the posterior vaginal fornix was performed. Species identification of microorganisms was performed by the method of direct protein profiling with the help of MALDI-TOF mass spectrometry, FLEX series, Bruker Daltonic GmbH, Germany. Results. The growth of flora was obtained in 761 patients (95%). Bacterial vaginosis (n = 71), Candida vulvovaginitis (n = 83) were diagnosed in 154 patients (19%). Lactobacillus spp. were found in 80% (n = 637) of examined women, of them in 161 (20%) in monoculture. Bacterial carriage was diagnosed in 55.8% of cases. The prevalence of Enterococcus faecalis, Escherichia coli, Streptococcus agalactiae, Staphylococcus epidermidis and Staphylococcus haemoliticus was noted. They were much more rarely detected in monoculture: E. faecalis (1%), S. agalactiae (0.1%), S. epidermidis (0.1%), Candida albicans (0.1%). In 39 (5%) patients, no growth of flora was found. In 17 patients we found extended-spectrum B-lactamase-producing bacteria: E. coli (n = 15), Klebsiella pneumoniae (n = 2). Conclusion. Taking into account a high prevalence of vaginal bacterial carriage in pregnant women in the 3rd trimester, it might be expedient to consider inclusion of microbiological examination of vaginal discharge in the basic spectrum of antenatal observation and screening examination of pregnant in the Russian Federation.
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The in vitro examination of the effectiveness of antiseptic substances for a surface disinfection of teeth inoculated with Escherichia coli
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01.01.2018 |
Makeeva I.
Franko A.
Semenov A.
Byakova S.
Novozhilova N.
Dezhurko-Korol V.
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Stomatologiia |
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The aim of the study was to assess the possibility of penetration of Escherichia coli bacteria into the dentinal tubules of the samples and determine the antimicrobial efficacy after 5 and 20 min exposition of 3% H2O2 (groups I and II) and 1 and 5 min exposition of 3% NaOCl (groups III and IV) for surface disinfection of bovine teeth. The samples were subjected to inoculation with E. coli suspension. The quality of disinfection was assessed with the three methods. The surface disinfection of samples proved to be effective only in groups II and IV. In the cultures of all dissolutions of dentinal chips suspensions in groups I, III and V there was a growth of E. coli in the form of a continuous pitch. In the group II a growth of Escherichia coli was revealed only in the initial dissolution in the quantity 1,8x101 CFU/ml, whereas in the group IV the growth was nil. Quantitative estimation of bacterial penetration using the method of maximum dissolutions revealed similar quantity of bacteria as in the group II as well as in the calculation of CFU. The application of 3% solution of H2O2 with the exposition of 20 minutes secures the qualitative surface disinfection of teeth without destruction of bacteria inside dental tubules and that allows to discover viable bacteria inside dentinal tubules.
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Elaboration of a bacterial cellulose matrix for the immobilisation of Escherichia coli cells
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01.01.2018 |
Gromovykh T.
Feldman N.
Tikhonova O.
Lutsenko S.
Timashev P.
Bardakova K.
Churbanov S.
Kiselyova O.
Kraeva M.
Grinevich A.
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International Journal of Nanotechnology |
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2 |
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© Copyright 2018 Inderscience Enterprises Ltd. This paper is concerned with the studies of a natural nanomaterial which is bacterial cellulose synthesised by Gluconacetobacter hansenii producer strain. It covers different fields of bacterial cellulose use, including medicine. The research has proved that bacterial cellulose matrices with immobilised cells have high potential as immobilisers of cells, including making probiotics of prolonged action. The matrices consisted of bacterial cellulose films were prepared by static cultivation of G. hansenii GH-1/2008 strain in the liquid medium. We have developed methods of washing out end toxins and producer cells of the films in the solutions of sodium bicarbonate, sodium dodecyl sulphate, and sodium hydroxide. The LAL-test has revealed that washing the films with sodium dodecyl sulphate is more efficient. By means of electron scanning and atomic force microscopy (AFM), we have determined that bacterial cellulose matrices have a layered structure, smooth surface, and adhesion of E. coli test strain cells. The adhesive capacity, the energy of adhesion and contact angle is higher for 50 um thick films than for 20 um thick ones. The bacterial cellulose matrices obtained by the biosynthesis of G. hansenii strain can be recommended for the immobilisation of different producer cells.
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