[Impairments in metabolism of amino acids - precursors of gasotransmitters - in the premature birth]. / Narushenie obmena aminokislot ­ predshestvennikov gazotransmitterov pri prezhdevremennykh rodakh.

The content of amino acids - sources of gasotransmitters and the activity of enzymes of their metabolism have been studied in the placenta and amniotic fluid in full-term and complicated by preterm birth (PB). Determination of amino acids was carried out using an automatic analyzer; specific spectrophotometric methods were used to assess the activity of enzymes. The development of PB is accompanied by changes in the amino acid level already in the second trimester of pregnancy. Correlation of differently directions was found between the level of amino acids and the activity of the corresponding enzymes. The imbalance of amino acids in the fetoplacental system in PB is accompanied by a change in the production of their low molecular weight derivatives gas transmitters (NO, CO, H2S), which play an important role in the regulation of numerous metabolic processes.

[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.

[The activity of enzymes of amino acid metabolism of the placenta in different terms of the physiological and complicated pregnancy.]

The activity of amino acid metabolism enzymes and the content of free amino acids in the placenta during physiological pregnancy and placental insufficiency (PI) were studied using spectrophotometric methods and ion-exchange chromatography. It was found that in PI placental activity of the studied enzymes: alanine-, cysteine-e, tyrosine-, glutamino- transferase, glutathione synthetase, glutamate dehydrogenase decreases at different periods of gestation. The opposite variations occur for aspartataminotranferase and glutaminase. Similar changes are detected for amino acids synthesized or used in the course of appropriate reactions: aspartic acid, glutamic acid, glutamine, alanine, cysteine, tyrosine, arginine. The correlation between enzyme activity and amino acid content was revealed. Different periods of pregnancy are characterized by varying degrees of change, especially expressed in the second trimester, characterized by the most intense growth and development of the fetus, and its increased needs for trophic material. The revealed changes obviously play a pathogenetic role in the formation and further development of PI.

[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.

[Disbalance of redox processes in amniotic fluid at pre-eclampsia.]

The parameters of pro and antioxidant processes in the amniotic fluid in the dynamics of physiological pregnancy and preeclampsia were studied. Spectrophotometric and fluorimetric methods were used to evaluate the activity of NADPH-oxidase, xanthine-oxidase, superoxide dismutase (SOD), catalase, glutathione-dependent enzyme systems, the content of antioxidant vitamins - retinol and α-tocopherol, lipid peroxidation products (LPP) - diene conjugates and Schiff bases, and parameters of peroxide chemiluminescence. It was found that with preeclampsia there is an increase in the activity of prooxidant enzymes NADPH-oxidase and xanthine-oxidase, a decrease in the activity of antioxidant enzymes SOD, catalase, glutathione peroxidase, glutathione reductase, glutathione transferase, and the content of fat-soluble vitamins. Between pro- and antioxidant indices and the content of LPP, intensity of chemiluminescence a correlation is established, the degree of which is different in the II and III trimesters of pregnancy. The revealed changes obviously play a pathogenetic role in the formation and further development of preeclampsia, and the indicators of oxidation-reduction processes can serve as informative markers of pre- and postnatal pathology.

[Influence of amino acid imbalance in maternal and fetal organisms on the development of placental insufficiency and the course of the neonatal period.]

The content of amino acids in maternal serum and umbilical cord blood during physiological pregnancy and placental insufficiency (PI) was estimated using ion-exchange chromatography. It was found that that the content of arginine, serine, glutamine, alanine, cysteine, methionine, tryptophan, leucine, phenylalanine and proline, which are involved in many metabolic processes necessary to maintain the normal functioning of the mother-placenta-fetus system, was decreases in maternal serum at PI. Opposite deviations have dicarboxylic amino acids, lysine, histidine, glycine, tyrosine and threonine, the number of which increases, strengthening the nitrogen imbalance. In the blood serum of the umbilical cord for most amino acids, was a similar direction of changes, but the degree of it is more pronounced. The opposite direction of changes is found for glycine, threonine, tyrosine, the content of which is reduced. The results of the study make it possible to broaden the understanding of the mechanisms of PI development and to offer informative tests for predicting the state of newborns.

[Impairments of placental amino acid metabolism in fetal growth restriction]. / Narushenie platsentarnogo obmena aminokislot pri zaderzhke rosta ploda.

The content of the amino acids in the placenta during physiological pregnancy and fetal growth restriction (FGR) has been investigated my means of the method of ion-exchange chromatography. It has been found that in FGR the placental amino acid pool is characterized by a decreased content of arginine, proline, alanine, serine, cysteine, methionine, tryptophan, leucine, threonine, tyrosine, phenylalanine, glutamine and an increased content of dicarboxylic amino acids, lysine, histidine and glycine. These changes are accompanied by altered activity of some enzymes of amino acid metabolism, and the degree of these changes correlates with the level of corresponding amino acids.

Proteomic Profiling of the Blood Serum for Prediction of Premature Delivery.

Mass-spectrometric profiling of the serum in women at weeks 16-17 of gestation was carried out in order to detect proteomic predictors of preterm delivery. Changes in the production of 25 proteins (down-regulation for 13 proteins and up-regulation for 12 proteins) were detected in the sera of women whose pregnancies eventuated in premature deliveries. Among them, proteins with various regulatory functions were distinguished: antioxidant enzymes, chaperons, cytoskeleton proteins, cell adhesion molecules, and proteins involved in angiogenesis, proteolysis, transcription, inflammation processes, binding and transportation of various ligands. These results indicated the formation of proteomic imbalance as early as during trimester II, this eventually leading to premature delivery. The detected serum proteins were suggested as markers for early prediction of premature delivery.

[The imbalance of metal-containing proteins and free metal ions in the amniotic fluid during fetal growth].

The levels of zinc, copper, iron, and magnesium ions, and some of their binding proteins have been investigated in an amniotic fluid under the fetal growth retardation (FGR). FGR, developed under conditions of placental insufficiency, is characterized by a decrease in the content of zinc, iron, and magnesium ions and by an increase in the copper content in the amniotic fluid in the II and III trimesters of pregnancy. During these trimesters the levels of ceruloplasmin, ferritin, and Ca2+,Mg2+-ATPase were lower in FGR, while the level of zinc-a-2-glycoprotein was higher than during the same periods of normal pregnancy. Changes in the parameters studied in the amniotic fluid were associated with developmental disorders of the newborns. These changes obviously have a pathogenetic importance in the development of FGR, and the levels of metal ions and their ratio in the amniotic fluid can be used as markers of the pre- and postnatal pathology.

[Imbalance of system of glutamin - glutamic acid in the placenta and amniotic fluid at placental insufficiency].

Metabolism of glutamine and glutamic acid has been investigated in the placenta and amniotic fluid under conditions of placental insufficiency. The development of placental insufficiency is characterized by the increased content of glutamic acid and a decrease of glutamine in both placenta and amniotic fluid. These changes changes were accompanied by changes in the activity of enzymes involved in the metabolism of these amino acids. There was a decrease in glutamate dehydrogenase activity and an increase in glutaminase activity with the simultaneous decrease of glutamine synthetase activity. The compensatory decrease in the activity of glutamine keto acid aminotransferase did not prevent a decrease in the glutamine level. The impairments in the system glutamic acid-glutamine were more pronounced during the development of premature labor.

[Amino acid composition of amniotic fluid during physiological and complicated pregnancy].

An ion exchange chromatographic technique was used to study the content of 18 free amino acids in the amniotic fluid of women with physiological and complicated pregnancy. The amino acid spectrum of amniotic fluid in placental insufficiency was found to be characterized by the lower content of arginine, serine, glycine, alanine, cysteine, tyrosine, phenylalanine, and glutamine, which are involved in many metabolic processes required to maintain biochemical homeostasis during gestation. There was an opposite tendency for dicarboxylic amino acids and histidine, the levels of which increased, by enhancing nitrogen imbalance. The degree of the found impairments of a number of amino acids considerably increased in case of birth of babies with brain lesions, which permits use of their amniotic fluid determination to predict the neonatal status.

New approaches to molecular diagnostics of prenatal pathology.

Analysis of the spectrum of amniotic fluid proteins in physiological and abnormal pregnancy using proteomic analysis allowed detection of a number of difference proteins, that are absent or, alternatively, appear in gestosis. Among absent proteins, there were NADPH-dependent carbonyl reductase, epidermal fatty acid-binding protein, haptoglobin, calgranulins A and B. In contrast to proteomic spectrum of amniotic fluid in physiological pregnancy, 7 new proteins appear during gestosis, 3 of them were identified: C area of immunoglobulin K-chain, breast cancer metastasis suppressor-1, and protein-1 containing AIG2-like domain. Possible effects of revealed differences in proteomic spectrum on development of main disturbances during gestosis are discussed. Difference proteins detected in amniotic fluid may serve as gestosis markers.

[Proteins-markers of placental insufficiency].

The proteomic analysis of the amniotic fluids of women with physiological pregnancy and pregnancy, complicated with placental insufficiency has been carried out on the II and III trimesters. The following difference in protein patterns have been recognized: i) appearance of several proteins lacking in physiological pregnancy; ii) absence of several proteins detectable during physiological pregnancy--hippocalcin-like protein 1, CDC37-like protein, NKG2D ligand 2 (II trimester), CDC37-like protein, NKG2D ligand 2 (III trimester). The established differences in the amniotic fluid spectrum, obviously, have the pathogenetic meaning in the placental insufficiency development. The revealed proteins of distinction may serve as markers of this obstetrical pathology.

Phosphorylation and carbonylation of placental proteins in normal pregnancy and gestosis.

Study of posttranslational changes in placental proteins revealed disorders in the intensity of their phosphorylation and carbonylation in patients with placental failure. Phosphorylation was reduced for the majority of endogenous placental proteins, substrates for cAMP- and cGMP-dependent protein kinases. An opposite dynamics was noted for oxidative modification of proteins. The content of carbonyl derivatives evaluated in spontaneous and metal-catalyzed oxidation of placental proteins was elevated in gestosis in comparison with the normal level.

[Production and reception of growth factors in the placenta during physiological and gestosis complicated pregnancy].

30 women with physiological pregnancy and 28 women with gestosis were examined. In the early chorion obtained after abortion and on the full-term placenta the content of the epidermal growth factor (EGF), vascular-endothelial growth factor (VEGF) and their receptors were studied by means of the ELISA method. In the process of normal gestation the increase of the placental production both of the EGF and VEGF was found. During the pregnancy complicated with gestosis and miscarriage in the first trimester the content of EGF and its receptor was lower compared to the physiological values. For VEGF and its receptor opposite changes were found: the increase of quantity of the growth factor and the decrease of its receptor. In the case of gestosis and term pregnancy the content of the both growth factors and their receptors was lower than in controls. The revealed changes in production of the angiogenic growth factors and their receptors in the placenta may have the pathogenic importance in the development of gestosis.

Production of vascular endothelial growth factor and endothelin in the placenta and umbilical cord during normal and complicated pregnancy.

The concentrations of vascular endothelial growth factor and endothelin in the placenta progressively increased during normal pregnancy. Production of vascular endothelial growth factor and endothelin in the placenta exceeded the normal during trimester I miscarriage and trimester III premature birth accompanied by intrauterine hypoxia. The concentration of these vasoactive substances during premature birth also increased in the umbilical cord. The compensatory decrease in the concentrations of vascular endothelial growth factor and endothelin in the placenta and umbilical cord was observed during full-term pregnancy with threatened abortion.

[The influence of nitric oxide generation on the production of interleukins and processes of prostaglandin synthesis in fetal membranes in spontaneous parturition and uterine inertia]. / Vliianie generatsii oksida azota na produktsiiu interleikinov i protsessy sinteza prostaglandinov v plodnykh obolochkakh pri spontannoi rodovoi deiatel'nosti i slabosti rodovykh sil.

62 women in 38-40 weeks gestation were examined, 37 of them had normal delivery, whereas others (25) with uterine inertia, had delivery by Cesarean section. In the fetal membranes preparations nitric oxide production, interleukins -1beta and -6, arachidonic acid content and phospholipase A2 activity (limiting the rate of prostaglandin biosynthesis) were examined. In the case of uterine inertia increase of nitric oxide generation, decrease of interleukins and arachidonic acid levels and phospholipase A2 activity was found. Modulation of nitric oxide production with inhibitors and activators allowed to emphasize the role of NO in regulation of cytokine response and control of initial stages of prostaglandins synthesis.

[Prognostic value of neuraminidase activity in mammary gland secretion in fetal death]. / Prognosticheskoe znachenie opredeleniia aktivnosti neiraminidazy v sekrete molochnukh zhelez pri vnutriutrobnoi gibeli ploda.

Neuraminidase activity and levels of sialic acids and some hormones were measured in mammary secretion of women with induced pregnancy with a history of endocrine sterility. The activity of neuraminidase in the breast milk of pregnant women with galactorrhea may serve an informative prognostic test indicating intrauterine fetal death for 8-10 days.

[Use of chemiluminescent analysis of the blood in prognostication of the course of adaptation period of the newborn]. / Ispol'zovanie khemiliuminestsentnogo analiza krovi dlia prognozirovaniia techeniia adaptatsionnogo perioda u novorozhdennykh.

Study of spontaneous and hydrogen peroxide-induced chemiluminescence of the blood serum of children born clinically healthy to mothers with complicated course of pregnancy showed that an increase of the total photosum of spontaneous chemiluminescence on day 3 of life may be regarded as a criterion of failure of adaptation; other criteria are the photosum value and intensity of the amplitude of flashes of induced chemiluminescence of newborn blood sera over the entire course of the neonatal period.