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Article: Adaptation of Microarray Assay for Serum Amyloid a Analysis in Human Serum.

Smoldovskaya, O V / Voloshin, S A / Novikov, A A / Aleksandrova, E N / Feyzkhanova, G U / Rubina, A Yu

2022 Volume 56, Issue 2, Page(s) 290–296

Abstract: Serum amyloid A is an inflammatory biomarker whose concentration changes during infectious and inflammatory diseases. SAA's tendency for aggregation and complex formation makes it difficult to determine its concentration in samples, especially when there ...

Abstract	Serum amyloid A is an inflammatory biomarker whose concentration changes during infectious and inflammatory diseases. SAA's tendency for aggregation and complex formation makes it difficult to determine its concentration in samples, especially when there is an increased level of it. Immunofluorescence SAA determination on a microarray was adapted for SAA quantification in human serum. Both the procedure and the diluent for the calibrator samples were chosen to obtain a dynamic range between 1 and 100 μg/mL. Mixtures of animal (rabbit, goat, mouse) sera with recombinant antigen diluted in certain concentrations were used for the calibrator samples. The method was tested using serum samples from 15 patients with rheumatoid arthritis or ankylosing spondylitis and 9 healthy donors. The results obtained on the microarray demonstrated a good correlation with the results determined by ELISA (Pearson's correlation coefficient is 0.93). The method developed could be a convenient tool for assessing SAA levels in a number of diseases, such as rheumatoid arthritis or infections of various etiologies, characterized by a significant increase in the level of this protein in the blood. The use of a microarray for the analysis allows the determination of the SAA concentration simultaneously with other inflammatory biomarkers.
Language	English
Publishing date	2022-04-14
Publishing country	Russia (Federation)
Document type	Journal Article
ZDB-ID	213541-3
ISSN	1608-3245 ; 0026-8933
ISSN (online)	1608-3245
ISSN	0026-8933
DOI	10.1134/S0026893322020145
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Article: [Adaptation of Microarray Assay for Serum Amyloid A Analysis in Human Serum].

Smoldovskaya, O V / Voloshin, S A / Novikov, A A / Aleksandrova, E N / Feyzkhanova, G U / Rubina, A Yu

Molekuliarnaia biologiia

2022 Volume 56, Issue 2, Page(s) 336–342

Abstract	Serum amyloid A is an inflammatory biomarker whose concentration changes during infectious and inflammatory diseases. SAA's tendency for aggregation and complex formation makes it difficult to determine its concentration in samples, especially when there is an increased level of it. Immunofluorescence SAA determination on a microarray was adapted for SAA quantification in human serum. Both the procedure and the diluent for the calibrator samples were chosen to obtain a dynamic range between 1 and 100 μg/mL. Mixtures of animal (rabbit, goat, mouse) sera with recombinant antigen diluted in certain concentrations were used for the calibrator samples. The method was tested using serum samples from 15 patients with rheumatoid arthritis or ankylosing spondylitis and 9 healthy donors. The results obtained on the microarray demonstrated a good correlation with the results determined by ELISA (Pearson's correlation coefficient is 0.93). The method developed could be a convenient tool for assessing SAA levels in a number of diseases, such as rheumatoid arthritis or infections of various etiologies, characterized by a significant increase in the level of this protein in the blood. The use of a microarray for the analysis allows the determination of the SAA concentration simultaneously with other inflammatory biomarkers.
MeSH term(s)	Animals ; Arthritis, Rheumatoid/diagnosis ; Arthritis, Rheumatoid/genetics ; Biomarkers ; Enzyme-Linked Immunosorbent Assay ; Goats ; Humans ; Mice ; Rabbits ; Serum Amyloid A Protein/analysis ; Serum Amyloid A Protein/genetics ; Serum Amyloid A Protein/metabolism ; Spondylitis, Ankylosing
Chemical Substances	Biomarkers ; Serum Amyloid A Protein
Language	Russian
Publishing date	2022-04-11
Publishing country	Russia (Federation)
Document type	Journal Article
ZDB-ID	213542-5
ISSN	0026-8984
ISSN	0026-8984
DOI	10.31857/S0026898422020173
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Article: Analysis of rheumatoid factor and acute phase proteins using microarrays in patients with rheumatoid arthritis.

Feyzkhanova, G U / Voloshin, S A / Novikov, A A / Aleksandrova, E N / Smoldovskaya, O V / Rubina, A Yu

Klinicheskaia laboratornaia diagnostika

2022 Volume 67, Issue 1, Page(s) 43–47

Abstract: One of the biomarkers of biggest clinical importance in rheumatoid arthritis (RA) is rheumatoid factor (IgM RF). The rheumatoid factor has insufficient sensitivity and specificity, therefore, to increase the diagnostic information of the test, acute ... ...

Title translation	Определение ревматоидного фактора и белков острой фазы воспаления на биочипах у пациентов с ревматоидным артритом.
Abstract	One of the biomarkers of biggest clinical importance in rheumatoid arthritis (RA) is rheumatoid factor (IgM RF). The rheumatoid factor has insufficient sensitivity and specificity, therefore, to increase the diagnostic information of the test, acute phase proteins were used as concomitant biomarkers. Using biological microchips, we measured IgM RF, C-reactive protein (CRP) and Serum amyloid protein A (SAA) in patients with RA (n = 60), ankylosing spondylitis (AS) (n=55), systemic lupus erythematosus (SLE) (n=20) and healthy donors (HD) (n=9). It was shown that the medians of IgM RF concentrations are significantly higher (p<0.01) in patients with RA compared to patients suffering from other diseases and healthy donors. CRP and SAA were also significantly increased (p<0.05) in patients with RA and AS compared with SLE and HD. It has been shown that the complex determination of three biomarkers in differentiating RA patients with the comparison group had a higher diagnostic sensitivity than the isolated determination of IgM RF, while the addition of SAA makes the greatest contribution to improving the diagnostic characteristics of the biomarker panel: the use of a logistic regression model based on IgM RF and SAA allowed to increase the diagnostic sensitivity of the analysis from 58.3% to 65%. Thus, the developed microarray-based method can be used to detect and elucidate the diagnostic characteristics of RA biomarkers; however, further use requires validation of the obtained results on an expanded sampling.
MeSH term(s)	Acute-Phase Proteins ; Arthritis, Rheumatoid/diagnosis ; Arthritis, Rheumatoid/genetics ; Biomarkers ; C-Reactive Protein ; Humans ; Lupus Erythematosus, Systemic ; Rheumatoid Factor
Chemical Substances	Acute-Phase Proteins ; Biomarkers ; C-Reactive Protein (9007-41-4) ; Rheumatoid Factor (9009-79-4)
Language	English
Publishing date	2022-01-25
Publishing country	Russia (Federation)
Document type	Journal Article
ZDB-ID	1155086-7
ISSN	0869-2084
ISSN	0869-2084
DOI	10.51620/0869-2084-2022-67-1-43-47
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Article: [Microarray for Quantitative Determination of Inflammatory Biomarkers in a Culture Medium].

Voloshin, S A / Feyzkhanova, G U / Savvateeva, E N / Smoldovskaya, O V / Rubina, A Yu

Molekuliarnaia biologiia

2020 Volume 54, Issue 6, Page(s) 1046–1056

Abstract: Cytokines and acute phase proteins play an important role in the development of the immune response during inflammatory reactions. Depending on the type of disease, the development of inflammation is accompanied by changes in concentrations (both ... ...

Abstract	Cytokines and acute phase proteins play an important role in the development of the immune response during inflammatory reactions. Depending on the type of disease, the development of inflammation is accompanied by changes in concentrations (both decrease and increase) of not one, but many inflammatory biomarkers. Here, a quantitative microarray-based method for multiplex immunoassay of eight biomarkers of human inflammation, namely acute phase proteins (C-reactive protein, serum amyloid protein A) and cytokines (IL-6, IL-8, IL-17, IL-18, IP10/CXCL10, TNFα) was developed and the possibility of its use for the detection of inflammatory biomarkers in a culture medium has been demonstrated. The developed method can be used to evaluate changes of the inflammatory biomarker profile induced by different agents or to determine the concentrations of biomarkers after activation of cells while studying different diseases with the help of in vitro models.
MeSH term(s)	Biomarkers ; Culture Media ; Cytokines/analysis ; Cytokines/genetics ; Humans ; Inflammation ; Inflammation Mediators/analysis ; Microarray Analysis
Chemical Substances	Biomarkers ; Culture Media ; Cytokines ; Inflammation Mediators
Language	Russian
Publishing date	2020-12-02
Publishing country	Russia (Federation)
Document type	Journal Article
ZDB-ID	213542-5
ISSN	0026-8984
ISSN	0026-8984
DOI	10.31857/S0026898420060130
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Article: [Modification of Anti-Glycan IgG and IgM Profiles in Allergic Inflammation].

Butvilovskaya, V I / Smoldovskaya, O V / Feyzkhanova, G U / Filippova, M A / Pavlushkina, L V / Voloshin, S A / Rubina, A Yu

Molekuliarnaia biologiia

2018 Volume 52, Issue 4, Page(s) 634–643

Abstract: Glycans and anti-glycan antibodies (AGAs) are essential for infiltration of inflammatory cells in various allergies. The glycocalyx structure of the cells is modified during disease progression, and this modification is possible to evaluate by assessment ...

Abstract	Glycans and anti-glycan antibodies (AGAs) are essential for infiltration of inflammatory cells in various allergies. The glycocalyx structure of the cells is modified during disease progression, and this modification is possible to evaluate by assessment of AGAs. A printed glycan array with 55 immobilized glycans and immobilized antibodies to IgG, IgA, and IgM was used to study the changes in AGA profiles in bronchial asthma (BA). Levels of antibodies to certain glycans in BA patients statistically differed from levels in healthy donors (p < 0.0007 by the Mann-Whitney test); the glycan set included 6Su-6`-SiaLec, Sia LeX, Sia6Htype2; Tαα, Manβ1-4GlcNAc, and Manα1-4Manβ. The obtained results help to better understand the mechanisms of the cell-mediated immune response in bronchial asthma and other types of allergic reactions.
MeSH term(s)	Adolescent ; Antibodies, Anti-Idiotypic/chemistry ; Antibodies, Anti-Idiotypic/immunology ; Antibodies, Immobilized/chemistry ; Antibodies, Immobilized/immunology ; Asthma/blood ; Asthma/immunology ; Asthma/pathology ; Child ; Child, Preschool ; Female ; Humans ; Hypersensitivity/blood ; Hypersensitivity/immunology ; Hypersensitivity/pathology ; Immunity, Cellular/immunology ; Immunoglobulin G/blood ; Immunoglobulin G/immunology ; Immunoglobulin M/blood ; Immunoglobulin M/immunology ; Inflammation/blood ; Inflammation/immunology ; Inflammation/pathology ; Male ; Polysaccharides/blood ; Polysaccharides/chemistry ; Polysaccharides/immunology
Chemical Substances	Antibodies, Anti-Idiotypic ; Antibodies, Immobilized ; Immunoglobulin G ; Immunoglobulin M ; Polysaccharides
Language	Russian
Publishing date	2018-08-15
Publishing country	Russia (Federation)
Document type	Journal Article
ZDB-ID	213542-5
ISSN	0026-8984
ISSN	0026-8984
DOI	10.1134/S0026898418040031
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Article: Cell-cell interactions in bacterial populations.

Voloshin, S A / Kaprelyants, A S

Biochemistry. Biokhimiia

2004 Volume 69, Issue 11, Page(s) 1268–1275

Abstract: In developing bacterial populations many essential processes, such as division, genetic transformation, sporulation, and synthesis of antibiotics and secondary metabolites, are regulated by intercellular communication mediated by secretion of signaling ... ...

Abstract	In developing bacterial populations many essential processes, such as division, genetic transformation, sporulation, and synthesis of antibiotics and secondary metabolites, are regulated by intercellular communication mediated by secretion of signaling molecules, such as homoserine lactones and peptides. Another intercellular communication type, namely a physical contact between cells (cell aggregation), plays a key role in formation of biofilms or cellular consortia and in cell proliferation under unfavorable conditions. The mechanisms involved in these two types of bacterial communication are discussed in this review.
MeSH term(s)	Bacterial Physiological Phenomena ; Cell Division ; Culture Media/metabolism ; Pheromones/physiology ; Signal Transduction
Chemical Substances	Culture Media ; Pheromones
Language	English
Publishing date	2004-06-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	1109-5
ISSN	1608-3040 ; 0006-2979 ; 0320-9717
ISSN (online)	1608-3040
ISSN	0006-2979 ; 0320-9717
DOI	10.1007/s10541-005-0072-9
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Article: [Hydrogel microchip as a tool for studying exosomes in human serum].

Butvilovskaya, V I / Tikhonov, A A / Savvateeva, E N / Ragimov, A A / Salimov, E L / Voloshin, S A / Sidorov, D V / Chernichenko, M A / Polyakov, A P / Filushin, M M / Tsybulskaya, M V / Rubina, A Yu

Molekuliarnaia biologiia

2017 Volume 51, Issue 5, Page(s) 817–823

Abstract: Exosomes are cell-derived vesicles that are secreted by both normal and cancer cells. Over the last decade, a few studies have revealed that exosomes cross talk and/or influence major tumor-related pathways such as angiogenesis and metastasis involving ... ...

Abstract	Exosomes are cell-derived vesicles that are secreted by both normal and cancer cells. Over the last decade, a few studies have revealed that exosomes cross talk and/or influence major tumor-related pathways such as angiogenesis and metastasis involving many cell types within the tumor microenvironment. The protein composition of the membrane of an exosome reflects that of the membrane of the cell of origin. Because of this, tumor-derived exosomes differ from exosomes that are derived from normal cells. The detection of tumor exosomes and analysis of their molecular composition hold promise for diagnosis and prognosis of cancer. Here, we present hydrogel microarrays (biochips), which contain a panel of immobilized antibodies that recognize tetraspanins (CD9, CD63, CD81) and prognostic markers for colorectal cancer (A33, CD147). These biochips make it possible to analyze the surface proteins of either isolated exosomes or exosomes that are present in the serum samples without isolation. These biochips were successfully used to analyze the surface proteins of exosomes from serum that was collected from a colorectal cancer patient and healthy donor. Biochip-guided immunofluorescent analysis of the exosomes has made it possible for us to detect the A33 antigen and CD147 in the serum sample of the colorectal cancer patient with normal levels of CEA and CA19-9.
Language	Russian
Publishing date	2017-09
Publishing country	Russia (Federation)
Document type	English Abstract ; Journal Article
ZDB-ID	213542-5
ISSN	0026-8984
ISSN	0026-8984
DOI	10.7868/S0026898417050081
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Article: [Cell aggregation in cultures of Micrococcus luteus studied by dynamic light scattering].

Voloshin, S A / Kaprel'iants, A S

Prikladnaia biokhimiia i mikrobiologiia

2005 Volume 41, Issue 6, Page(s) 647–651

Abstract: Cell aggregation was studied using the method of dynamic light scattering in the course of growth of Micrococcus luteus cultures in a liquid medium. The method detects particles ranging in size from 0.5 to 1000 microm in samples containing no more than ... ...

Abstract	Cell aggregation was studied using the method of dynamic light scattering in the course of growth of Micrococcus luteus cultures in a liquid medium. The method detects particles ranging in size from 0.5 to 1000 microm in samples containing no more than 10(5) cells/ml. When grown in liquid media, M. luteus forms aggregates; during the lag phase, 80% of the cells are found in aggregates of 10 to 1000 microm, only minor amounts being represented by single cells. With the onset of exponential growth, the aggregates were decomposed, and single cells became prevalent in the culture liquid. This observation confirms that the aggregation of the cells during the lag phase is prerequisite to the initiation of bacterial growth. The method may be used in biotechnology for monitoring the state of bacterial cultures.
MeSH term(s)	Culture Media ; Lasers ; Micrococcus luteus/cytology ; Micrococcus luteus/growth & development ; Micrococcus luteus/physiology ; Scattering, Radiation
Chemical Substances	Culture Media
Language	Russian
Publishing date	2005-11
Publishing country	Russia (Federation)
Document type	English Abstract ; Journal Article
ZDB-ID	412549-6
ISSN	0555-1099
ISSN	0555-1099
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Article: [The role of intercellular contacts in the initiation of growth and in the development of a transiently nonculturable state by the cultures of Rhodococcus rhodochrous grown in poor media].

Voloshin, S A / Shleeva, M O / Syroeshkin, A V / Kaprel'iants, A S

Mikrobiologiia

2005 Volume 74, Issue 4, Page(s) 489–497

Abstract: It was found that the growth of Rhodococcus rhodochrous cells in modified Saton's medium strongly depends on the rate of culture agitation in the flask: an agitation at 250 rpm in flasks with baffles stops cell multiplication, whereas slight agitation ... ...

Abstract	It was found that the growth of Rhodococcus rhodochrous cells in modified Saton's medium strongly depends on the rate of culture agitation in the flask: an agitation at 250 rpm in flasks with baffles stops cell multiplication, whereas slight agitation leads to pronounced culture growth. The growth retardation phenomenon was reversible and did not manifest itself in exponential-phase cultures or when the cells were grown in a rich medium; furthermore, it was not connected with the degree of culture aeration. When agitated at a moderate rate, the bacterial cells formed aggregates in the lag phase, which broke up into single cells in the exponential phase. The inhibitory effect of vigorous agitation was removed by the addition to the medium of the supernatant (SN) of a log-phase culture grown in the same medium with moderate agitation. Vigorous agitation is thought to interfere with the cell contacts, whose establishment is necessary for the development of an R. rhodochrous culture in a poor medium, which occurs in the form of (micro) cryptic growth. When grown in modified Saton's medium, R. rhodochrous cells were capable of transition, in the prolonged stationary phase, to a resting and transiently nonculturable state. Such cells could be resuscitated by incubation in a liquid medium with the addition of the supernatant or the Rpf secreted protein. The formation of transiently nonculturable cells was only possible under the conditions of a considerable agitation rate (250-300 rpm), which prevented secondary (cryptic) growth of the culture. This circumstance indicates the importance of intercellular contacts not only for the initiation of growth but also for the transition of the bacteria to a dormant state.
MeSH term(s)	Adaptation, Physiological ; Culture Media ; Rhodococcus/growth & development ; Rhodococcus/physiology ; Signal Transduction
Chemical Substances	Culture Media
Language	Russian
Publishing date	2005-07
Publishing country	Russia (Federation)
Document type	English Abstract ; Journal Article
ZDB-ID	209707-2
ISSN	0026-3656
ISSN	0026-3656
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Article: Proteins of the Rpf (resuscitation promoting factor) family are peptidoglycan hydrolases.

Telkov, M V / Demina, G R / Voloshin, S A / Salina, E G / Dudik, T V / Stekhanova, T N / Mukamolova, G V / Kazaryan, K A / Goncharenko, A V / Young, M / Kaprelyants, A S

Biochemistry. Biokhimiia

2006 Volume 71, Issue 4, Page(s) 414–422

Abstract: The secreted Micrococcus luteus protein, Rpf, is required for successful resuscitation of dormant "non-culturable" M. luteus cells and for growth stimulation in poor media. The biochemical mechanism of Rpf action remained unknown. Theoretical predictions ...

Abstract	The secreted Micrococcus luteus protein, Rpf, is required for successful resuscitation of dormant "non-culturable" M. luteus cells and for growth stimulation in poor media. The biochemical mechanism of Rpf action remained unknown. Theoretical predictions of Rpf domain architecture and organization, together with a recent NMR analysis of the protein structure, indicate that the conserved Rpf domain has a lysozyme-like fold. In the present study, we found that both the secreted native protein and the recombinant protein lyse crude preparations of M. luteus cell walls. They also hydrolyze 4-methylumbelliferyl-beta-D-N,N',N''-triacetylchitotrioside, a synthetic substrate for peptidoglycan muramidases, with optimum activity at pH 6. The Rpf protein also has weak proteolytic activity against N-CBZ-Gly-Gly-Arg-beta-naphthylamide, a substrate for trypsin-like enzymes. Rpf activity towards 4-methylumbelliferyl-beta-D-N,N',N''-triacetylchitotrioside was reduced when the glutamate residue at position 54, invariant for all Rpf family proteins and presumably involved in catalysis, was altered. The same amino acid substitution resulted in impaired resuscitation activity of Rpf. The data indicate that Rpf is a peptidoglycan-hydrolyzing enzyme, and strongly suggest that this specific activity is responsible for its growth promotion and resuscitation activity. A possible mechanism of Rpf-mediated resuscitation is discussed.
MeSH term(s)	Bacterial Proteins/chemistry ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Culture Media ; Cytokines/chemistry ; Cytokines/genetics ; Cytokines/metabolism ; Micrococcus luteus/cytology ; Micrococcus luteus/enzymology ; Micrococcus luteus/metabolism ; Mutagenesis, Site-Directed ; N-Acetylmuramoyl-L-alanine Amidase/chemistry ; N-Acetylmuramoyl-L-alanine Amidase/genetics ; N-Acetylmuramoyl-L-alanine Amidase/metabolism ; Peptide Hydrolases/chemistry ; Peptide Hydrolases/metabolism ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism ; Time Factors
Chemical Substances	Bacterial Proteins ; Culture Media ; Cytokines ; Recombinant Proteins ; resuscitation-promoting factor, bacteria ; Peptide Hydrolases (EC 3.4.-) ; N-Acetylmuramoyl-L-alanine Amidase (EC 3.5.1.28)
Language	English
Publishing date	2006-03-16
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1109-5
ISSN	1608-3040 ; 0006-2979 ; 0320-9717
ISSN (online)	1608-3040
ISSN	0006-2979 ; 0320-9717
DOI	10.1134/s0006297906040092
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