Proteomic analysis of blood serum - a new approach to the search for diagnostic markers of bronchial asthma in children.

Currently, bronchial asthma (BA) is one of the most pressing medical and social problems, the molecular aspects of the formation and development of BA are insufficiently studied and the diagnosis is not perfect. Carrying out proteomic analysis of BA will not only reveal new biomarkers specific to this disease, but also bring us closer to understanding its pathogenetic mechanisms. The purpose of the study: to study the proteomic profile of blood serum of children with BA to identify proteins associated with this disease A comprehensive clinical and laboratory examination of children suffering from BA and control group patients was performed. Proteomic analysis of depleted blood serum included high-resolution two-dimensional electrophoresis (1 direction: immobiline strips 17cm, pH 3-10, 2 direction: denaturing electrophoresis in 12.5% polyacrylamide gel), protein staining on gels with fluorescent dye Flamingo, protein identification by MALDI-TOF mass spectrometry using the search algorithm Mascot and the Swiss-Prot database. Comparison of the proteomic profile of BA serum and the control group patients serum allowed us to establish that the production of a number of proteins is reduced in this pathology. Among them, proteins in the molecular weight range of 16-33 kDa (p<0.05) were identified: glutathione peroxidase 3, transtyretin, complement components C4b and C3. Research shows that changes in the children's serum proteome occur in BA, affecting proteins that play an important role in immune responses, ligand transport, and antioxidant protection. Special attention should be paid to the differences identified in the course of this work (glutathione peroxidase, transtyretin, C3 and C4 fragments of the complement system) or their combinations. Studying the features of their expression will expand our understanding of the molecular mechanisms underlying chronic inflammation of this disease.

[Impairment of production and posttranslational changes of placental nuclear and membrane proteins with complicated pregnancy]. / Narushenie produktsii i posttransliatsionnye izmeneniia platsentarnykh iadernykh i membrannykh belkov pri oslozhnennoi beremennosti.

The content of nuclear and membrane proteins of the placenta, as well as posttranslational modification of these proteins in physiological pregnancy and placental insufficiency (PI) were studied. Differential centrifugation, electrophoresis in polyacrylamide gel, spectrophotometric methods were used. It was found that with PN there is a decrease in the degree of production of the studied proteins of varying degrees relative to control parameters. For chromatin proteins, a more pronounced decrease in the content of non-histone proteins was found in comparison with histones. Among histone fractions, the maximum decrease was detected in the H2A fraction. The degree of change in the amount of membrane proteins depends on the detergent used. Changes in posttranslational protein modifications disorders are characterized by a decrease in the content of amine and amide (especially difficult to hydrolyze) groups and an increase in carbonyl derivatives of proteins. The revealed changes in the composition and structure of the nuclear and membrane proteins of the placenta, performing numerous regulatory functions, can be triggering links in the chain of molecular damage in the placenta at PI.

[The imbalance of vasoactive components and arachidonic acid in the placenta and amniotic fluid in preeclampsia]. / Disbalans vazoaktivnykh komponentov i arakhidonovoi kisloty v platsente i okoloplodnykh vodakh pri preéklampsii.

The content of vasoactive compounds and arachidonic acid in the placenta and amniotic fluid was studied in full-term (39-40 weeks) physiological pregnancy and preeclampsia (PE). The content of metabolites of nitric oxide (NOx), endothelin-1, thromboxane B2 (TxB2), prostacycline (PGI2) and arachidonic acid was estimated using spectrophotometric, immunoenzyme methods and gas-liquid chromatography. It was found that in PE the content of vasoconstrictors, of endothelin and TxB2, increased in the placenta and amniotic fluid, while the content of vasodilators, PGI2 and NOx decreased. Despite the same directionality of changes in both studied objects, the degree of changes differed and was more pronounced in the placenta. A direct or inverse correlative relationship was found between various vasoactive components (depending on their effect on vascular tone). In the case of arachidonic acid changes in its content in PE correlated with the level of vasoactive compounds, the source of which it is. The revealed differences in the ratio of vasoactive components obviously play a pathogenetic role in the development of PE and its subsequent complications.

[Features of redox processes in the amniotic fluid at placental insufficiency]. / Osobennosti okislitel'no-vosstanovitel'nykh protsessov v okoloplodnykh vodakh pri platsentarnoi nedostatochnosti.

Activity of prooxidant enzymes (NADPH-oxidase and xanthine oxidase), antioxidant enzymes (superoxide dismutase (SOD) and catalase), enzymes of the glutathione-dependent systems, as well as antioxidant vitamins (retinol and a-tocopherol), lipid peroxidation products (LPP) (conjugated dienes and Schiff bases), and peroxide chemiluminescence were studied in the amniotic fluid at different periods of physiological pregnancy and placental insufficiency (PI). It was found that at PI the activity of NADPH-oxidase, xanthine oxidase increased and the activity of SOD, catalase, glutathione peroxidase, glutathione reductase, glutathione transferase and the content of fat-soluble vitamins decreased. The direct and inverse correlation between the studied pro- and antioxidant parameters and the content of LPP products, was found ro be different in the II and III trimesters of gestation. The revealed differences obviously reflect metabolic impairments in the fetoplacental complex, and the activity and level of the paremeters of redox processes can be used as tests for pre- and postnatal disorders.