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1.
Autoimmunity ; 51(6): 297-303, 2018 09.
Article in English | MEDLINE | ID: mdl-30369266

ABSTRACT

During NET formation, the content of neutrophils granules is released into the intercellular milieu. Consisting of many proteases and ROS species, formed NETs were shown to degrade cytokines (Schauer, Nat Med, 2014); while the content of neutrophil's azurophilic granules proved to contain glycosidases, secreted upon activation (Thaysen-Andersen, JBC, 2015), and formation of autoantibodies to neutrophil beta-glucoronidase was connected with the level of anti-MPO antibodies (Ab) (Martensson, Autoimmunity, 1992). Taking into account these facts, we aimed to investigate the possibility of NET-related changes in glycan composition on circulating IgG molecules and IgG-IgM immune complexes in multiple sclerosis (MS). This autoimmune disorder still has no reliable detection markers or established ways of treatment, besides widely accepted interferon therapy, making it a particularly interesting clinical condition. By applying capture lectin-ELISA, we analysed binding of α2,6 sialyl-specific lectins SNA, PSqL, and core α1,6-fucose specific lectin AAL to circulating IgG and related complexes in five groups of MS patients: untreated (17 persons); undergoing therapy with interferon (IFN) ß-1 b (15 persons), corticosteroids (methylprednisolone) (12 persons) and anti-B-cell monoclonal Ab (12 persons: Ocrelizumab, 6 persons and alemtuzumab, 6 persons). A group of 23 healthy donors served as control. Significant increase in neutrophil elastase activity, observed in the group of patients under corticosteroid treatment was also accompanied by sialyl-specific PSqL and SNA lectin binding to captured IgG molecules. Subsequent analysis demonstrated that sialic acid residues were exposed on free IgG and on circulating IgG-IgM immune complexes. Increased lectin binding was not observed for anti-myelin basic protein (one of the major autoAb in MS) Ab compared to total serum Ab. IFN therapy was accompanied by low neutrophil elastase activity and low amount of circulating immune complexes. Incubation of in vitro generated NETs with human serum revealed the digestion of high-molecular weight immune complexes with subsequent exposure of hidden glycoepitops. Obtained data indicate the potential of neutrophil-derived proteases to modify (partially degrade) circulating immune complexes leading to exposure of internal glycoepitops.


Subject(s)
Autoantibodies/blood , Extracellular Traps/enzymology , Glucuronidase/metabolism , Leukocyte Elastase/metabolism , Multiple Sclerosis/immunology , Adolescent , Adult , Aged , Autoantibodies/immunology , Epitopes/immunology , Epitopes/metabolism , Extracellular Traps/immunology , Female , Glucuronidase/immunology , Glycosylation , Humans , Immunoglobulin G/blood , Immunoglobulin G/immunology , Immunoglobulin M/blood , Immunoglobulin M/immunology , Immunosuppressive Agents/therapeutic use , Leukocyte Elastase/immunology , Male , Middle Aged , Multiple Sclerosis/blood , Multiple Sclerosis/drug therapy , Neutrophils/enzymology , Neutrophils/immunology , Young Adult
2.
Ukr Biochem J ; 86(4): 36-50, 2014.
Article in Ukrainian | MEDLINE | ID: mdl-25509182

ABSTRACT

Ovarian carcinoma is one of widely spread malignant diseases of female reproductive system. Mortality rate from it is much higher than from other female malignant diseases. During last 20 years the level of ovarian carcinoma in Ukraine and vast majority of other countries remains high manifesting no signs of decrease. This arouses interest of researchers to development of new methods of early diagnosis, therapeutic approach, prognostic criteria, especially biochemical ones, and means of prophylaxis of this pathology in medical scientific society. At present there are no specific diagnostic tests which would allow revealing the tumor on the initial stages of its development. In spite of vide arsenal of tumor markers, the only reliable test for ovarian carcinoma is determination of antigen CA-125. The results of basic modern research are discussed in this survey. They are aimed at finding out regulatory mechanisms connected with metabolism of L-arginine, activities of ATP-hydrolase systems and searching new markers of ovarian carcinoma. The main possible candidates for this role are determined.


Subject(s)
Biomarkers, Tumor/blood , Ovarian Neoplasms/diagnosis , Arginine/metabolism , CA-125 Antigen/blood , Female , Humans , Membrane Proteins/blood , Nitric Oxide/metabolism , Ovarian Neoplasms/metabolism , Sodium-Potassium-Exchanging ATPase/metabolism
3.
Biosens Bioelectron ; 22(12): 2790-5, 2007 Jun 15.
Article in English | MEDLINE | ID: mdl-17098416

ABSTRACT

A new formaldehyde-selective biosensor was constructed using NAD(+)- and glutathione-dependent recombinant formaldehyde dehydrogenase as a bio-recognition element immobilised on the surface of Si/SiO(2)/Si(3)N(4) structure. Sensor's response to formaldehyde was evaluated by capacitance measurements. The calibration curves obtained for formaldehyde concentration range from 10 microM to 20mM showed a broad linear response with a sensitivity of 31 mV/decade and a detection limit about 10 microM. It has been shown that the output signal decreases with the increase of borate buffer concentration and the best sensitivity is observed in 2.5mM borate buffer, pH 8.40. The response of the created formaldehyde-sensitive biosensor has also been examined in 2.5mM Tris-HCl buffer, and the shift to the positive bias of the C(V) curves along with the potential axis has been observed, but the sensitivity of the biosensor in this buffer is decreased dramatically to the value of 2.4 mV/decade.


Subject(s)
Aldehyde Oxidoreductases/metabolism , Biosensing Techniques/methods , Formaldehyde/analysis , Biosensing Techniques/instrumentation , Electric Capacitance , Hydrogen-Ion Concentration , Recombinant Proteins/metabolism , Semiconductors , Transducers
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