Histochemical Study of the Processes of Protein Oxidative Modification and Limited Proteolysis in the Endothelium of Myometrial Vessels in the Projection of the Utero-Placental Bed during Iron-Deficiency Anemia in Pregnancy

Utero-placental bed is the cumulation of gestationally altered endometrium at the place of ovum attachment to the uterine wall. As far as the protein oxidative modification and limited proteolysis in iron deficiency anemia are due to the fact that in conditions of hypoxia, free radical processes in the blood and tissues are enhanced, and iron deficiency is additionally able to cause hemodynamic disorders because of endothelial dysfunction in the vessels of the utero-placental area. The purpose of the study was to establish histochemical features of protein oxidative modification and limited proteolysis in the endotheliocytes of myometrial vessels in the projection of the utero-placental area depending on the degree of iron deficiency anemia in pregnant women. Materials and methods. By histochemical methods of Mikel Calvo, using reactions with bromophenol blue on "acidic" and "basic" proteins, and the method of A. Yasumа and T. Ichikawa, ninhydrin-Schiff reaction to free amino groups of proteins to assess the degree of limited proteolysis, in combination with computer microspectrophotometry and microdensitometry, quantitative characteristics of oxidative modification of proteins and limited proteolysis in endotheliocytes of myometrial segments of the utero-placental vessels in iron deficiency anemia of pregnant women were established. 74 biopsies of the observed uterine-placental area in physiological pregnancy and gestation based on iron deficiency anemia of I, II and III degrees of severity were investigated. Results and discussion. In physiological pregnancy, the intensity of oxidative modification of proteins and limited proteolysis is the lowest in endotheliocytes of myometrial segments of the spiral arteries and the highest one is in the endothelium of the vessels of the microcirculatory tract of the utero-placental area. In gestations based on iron deficiency anemia, intensification of proteins oxidative modification and limited proteolysis in the endothelium of all types of myometrial vessels of the projection of the utero-placental bed correlates with the severity of anemia. Iron deficiency anemia greatly affects the modification of proteins in the endothelial cells mostly of myometrial segments of spiral arteries of the placental bed. Conclusion. Intensification of protein oxidative modification and limited proteolysis in endotheliocytes of all type vessels of myometrium of the uterine-placental area formed by iron deficiency can be considered as a significant factor of endothelial dysfunction and a predictor of hemodynamic disorders of the placental bed

Histochemical Evaluation of the Processes of Protein Oxidative Modification in the Extravillous Cytotrophoblast of the Utero-Placental Bed during Iron-Deficiency Anemia in Pregnancy

Utero-placental bed is the cumulation of gestationally altered endometrium at the place of ovum attachment to the uterine wall. The key mechanism of this process is the cytotrophoblastic invasion. During iron deficiency anemia, an increase in the specific volume of the extravascular invasive trophoblast is taking place. Concern for the protein oxidative modification in iron deficiency anemia is due to the fact that in conditions of hypoxia, free radical processes in the blood and tissues are enhanced, and iron deficiency is additionally able to modify this problem. The purpose of the study was to establish the histochemical features of the processes of protein oxidative modification in the fractions of extravillous cytotrophoblast of the utero-placental bed depending on the degree of iron deficiency anemia in pregnant women. Material and methods. Quantitative characteristics of protein oxidative modification in the extravillous trophoblast of the utero-placental bed of pregnant women with iron deficiency anemia by means of the histochemical method using reactions with Bromophenol Blue on “acidic” and “basic” proteins according to Mikel Calvo method and computer microdensitometry. We studied 74 biopsies of the utero-placental bed of pregnant women with iron deficiency anemia of I, II and III degrees. The term of gestation was 37-40 weeks. Results and discussion. During physiological pregnancy, the ratio between "acidic" and "basic" proteins in trophoblast cells, even normally, is characterized by a predominance of "acidic" proteins, and evenly in both intravascular and extravascular fractions of cytotrophoblast. Intensification of processes of protein oxidative modification in the cytotrophoblast of the utero-placental bed during iron deficiency anemia of I-II degrees can be assessed as moderate, with an increase in the cells of the endothelium-replacing fraction of cytotrophoblast compared with the interstitial. In conditions of anemia of III degree, a significant predominance of "acidic" proteins in the intravascular cytotrophoblast was noted. Conclusion. During the physiological pregnancy, the intensity of protein oxidative modification was equal in all fractions of the extravillous cytotrophoblast in the utero-placental bed. In the case of gestation with iron deficiency anemia, significant intensification of the protein oxidative modification in the extravillous cytotrophoblast correlated with the severity of anemia. Background iron deficiency anemia significantly affected the processes of protein oxidative modification in the endothelium-replacing cytotrophoblast

Estimation of efficiency of complex cytoprotective therapy of pregnant women with fetal growth delay

The objective: estimate efficiency and influence of complex, cytoprotective therapy of pregnant women with fetal growth delay on biochemical indexes and results of delivery. Materials and methods. 93 women with monocyesis at term of 28-34 gestation weeks took part in the research. Group І includes 30 pregnant women with fetal growth delay (FGD). The pregnant women of the present group were prescribed with complex, cytoprotective therapy. It includes prescription of thiotriazolin in complex with L-arginine and diosmin. Group ІІ is presented by 33 pregnant women with FGD whose management of pregnancy and delivery is provided by valid orders of Ministry of Healthcare of Ukraine. Group ІІІ (the control one) consists of 30 pregnant women without FGD. Research of protein oxidative modification (POM) markers and level of stable nitrogen oxide metabolites (NO) were estimated in blood serum with spectrophotometric method. Reduced glutathione (GSH) was determined with calculation of its level according to the calibration curve. Results. Primary estimation of their content showed statistically valid (p < 0.05) lower GSH and NO in comparison of indexes of pregnant women without FGD (ІІІ group) with indexes of pregnant women with FGD (both І and ІІ groups). This fact indicates decrease of antioxidant protection and deficit of the main vessel dilator in the present group. Further estimation of biochemical indexes in dynamics of pregnancy permitted to determine considerable progress of imbalance between oxidants and antioxidants and absence of significant changes of NO level among pregnant women of II group. In pregnant women of I group who got the proposed complex cytoprotective therapy, on contrary, statistically valid (p < 0.05) decrease of POM indexes together with increase of GSH and NO level that indicates its positive influence were found. By means of analysis of special features of the pregnancy course in research group there was stated that the rate of habitual pregnancy non-carrying (HPN) in group II made 39.4% and considerably exceeded relevant indexes of І (20.0%) and ІІІ (16.7%) groups. Hypertensive disorders during pregnancy and labour are diagnosed only in pregnant women of І (6.7%) and ІІ (9.1%) groups. Average index of giving birth in І and ІІІ groups made 38.5±0.6 weeks and 39.4 ± 0.5 weeks correspondingly and in group ІІ it was lower 37.2±0.6 weeks. Index of preterm delivery (PD) rate in І group made 3.3% and was 6 times lower than appropriate index of ІІ group (21.2%). PD rate was absent in ІІІ group. Rate of the fetus distress in ІІ group (18.2%) was 2.7 times higher than the relevant index of І group (6.7%) and in ІІІ group it made 3.3%. Condition of newborns in research groups was estimated under Apgar scale. Both at the 1-st and 5-th minute the general index under appropriate scale was statistically valid (p < 0.05) and lower than in ІІ group as compared to indexes of І and ІІІ groups. During description of perinatal complications there was stated considerable percentage of neonatal encephalopathy and jaundice in ІІ group, namely 33.3% and 36.4% correspondingly. In І group their rate made 10.0% and 3.3%, correspondingly. In structure of perinatal pathology prematurity was in 21.2% of newborns of ІІ group that is 6 times higher than appropriate index of І group (3.3%). Mentioned complications were not determined in ІІІ group. In calculation under weight and height parameters of newborns the rate of FGD in ІІ group was the highest and made 88.6%. Mentioned index in І and ІІІ groups made 60.0% and 10.0%, correspondingly. Conclusion. The proposed complex cytoprotective therapy including combined use of thiotriazolin, L-arginine and diosmin (that considerably increases anti-ischemic, antioxidant and endothelium protective action of complex therapy) in women with fetal growth delay has the positive effect on oxidative-reductive hemeostasis and thiol-disulfide balance of mother-placenta-fetus system. Obstetric and perinatal results of delivery in the present group of pregnant women are characterized by statistically valid (р < 0.05) prevalence of estimation indexes under Apgar scale and correspondingly processes of adaptation as well as results of anthropomorphic measurements of newborns, their weight and height parameters and lower rate of neonatal encephalopathy and delivery of premature babies. Keywords: pregnancy, fetus growth inhibition, protein oxidative modification, antioxidant defense system, oxidative stress, obstetric and perinatal complications, cytoprotective therapy.

Local antioxidant effect of original dermal film with melatonin in thermal injury

Oxidative stress (OS) escalation associated with thermal trauma (TT) and pleiotropic effects of melatonin (MT) suggest a study of protective properties of the latter when applied as part of a novel dermal film (DF) to skin burns. This work aimed to assess the content of OS markers in the skin subjected to experimental TT and treated with DF with MT. Third A degree TT (area of 3.5%) were modeled by immersing a patch of skin in boiling water. Twelve cm2 of DF with 5 mg/g of MT were applied daily for 5 days. The parameters calculated were wound's area and epithelializatiohon rate. The products monitored in the burn wound were lipid peroxidation (LPO) products in heptane and isopropanol phases of the lipid extract and protein oxidative modification (POM) products, the modification being spontaneous and metal-dependent. With TT in the wound, the content of secondary and end LPO products in heptane and isopropanol phases increased on the 5th and 10th days; the total content of POM products grew on the 5th day (primary products, neutral) and on the 10th day (primary and secondary products, neutral). Application of DF to a TT wound reduced the burn area, increased the epithelialization rate (by the 10th day, the median went from 1.90% to 6.57%; p < 0.05), reduced the content of secondary and end LPO products in isopropanol phase (by the 10th day, the median went from 0.007 to 0.004 u.o.i; p < 0.05), reduced the total content of OMP products, namely that of primary neutral products — on the 5th day, of primary and secondary neutral products — on the 10th day. With TT present in the context of MT application, the burn area showed presence of secondary LPO products in heptane and isopropanol phases, LPO end products in isopropanol phase, POM products in the wound (basic and neutral primary/secondary POM products).

REDOX STATUS OF ASCITIC FLUIDAT DIFFERENT STAGES OF EXPERIMENTAL OVARIAN CANCER

In Russia, ovarian cancer (OC) is the first cause of mortality from vaginal cancer. Ascitic form of OC is considered to be the most malignant. The ascites composition is determined by the neoplasm biological properties in its development. The aim of the study was to evaluate the redox status ofascitic fluid at various stages of experimental OC. Materials and Methods. The work was performed on white outbred rats, 180–200 g, with a transplanted ascitic ovarian tumor. Ascites was taken from animals in the stationary and terminal stages of tumor growth. After removal of tumor cells, lipid peroxidation parameters were biochemically determined in it: diene conjugates, ketodienes, Schiff bases, malondialdehyde, levels of protein oxidative modification (346 nm – neutral aldehyde groups; 370 nm – neutral ketone groups; 430 nm – basic aldehyde groups; 530 nm – basic ketones) and antioxidant defense components – catalase and glutathione-S-transferase activity. The results were evaluated by means of Mann-Whitney U test (Stata 6.0). Results. As a result of the research conducted, the authors determined a significant reduction in the level of protein oxidative modification, a decrease in the catalase activity and a decrease in the malondialdehyde level at the terminal stage of tumor growth as compared with the stationary stage indices. Conclusion The redox status of ascitic fluid in experimental OC is characterized by a decrease in the oxidative modification of proteins, a decrease in the intensity of lipid peroxidation and a constant catalase and glutathione-S-transferase activity in the dynamics of neoplasm progression. Keywords: ovarian cancer, ascitic fluid, protein oxidative modification, lipid peroxidation, antioxidant system. В России рак яичников (РЯ) занимает первое место среди причин смертности от рака гениталий. Асцитная форма РЯ оценивается как наиболее злокачественная. Состав асцита при этом определяется биологическими свойствами неоплазмы в динамике ее развития. Целью исследования была оценка редокс-статуса асцитической жидкости на различных стадиях экспериментального РЯ. Материалы и методы. Работа выполнена на белых беспородных крысах массой 180–200 г с перевиваемой асцитной опухолью яичников. У животных в стационарную и терминальную стадии роста опухоли отбирался асцит, в котором после удаления опухолевых клеток биохимически определяли параметры перекисного окисления липидов: диеновые конъюгаты, кетодиены, основания Шиффа, малоновый диальдегид, уровни окислительной модификации белков (346 нм – альдегидные группы нейтрального характера; 370 нм – кетонные группы нейтрального характера; 430 нм – альдегидные группы основного характера; 530 нм – кетонные группы основного характера) и компоненты антиоксидантной защиты – активность каталазы и глутатион-S-трансферазы. Полученные результаты оценивались с использованием непараметрического критерия Манна–Уитни (Stata 6.0). Результаты. В результате проведенных исследований было установлено в терминальной стадии роста опухоли значимое по сравнению с показателями в стационарной стадии снижение уровня окислительной модификации белков, снижение активности каталазы и тенденция к снижению уровня малонового диальдегида. Заключение. Редокс-статус асцитической жидкости при экспериментальном РЯ характеризуется снижением окислительной модификации белков, тенденцией к снижению интенсивности перекисного окисления липидов и постоянной активностью каталазы и глутатион-S-трансферазы в динамике прогрессирования неоплазмы. Ключевые слова: рак яичников, асцитическая жидкость, окислительная модификация белков, перекисное окисление липидов, антиоксидантная система.

THE INFLUENCE OF THYROID HORMONES ON PROTEIN OXIDATIVE MODIFICATION IN CASE OF EXPERIMENTAL PERIODONTITIS

Introduction. The inflammatory-dystrophic process in periodontium is accompanied by its hypoxia, which causes the activation of free radical oxidation processes. Tissue destruction in periodontal diseases is considered to be the result of an altered inflammatory/immune response to microbial plaque and involves massive release of neutrophils, reactive oxygen species and enzymes. The aim of the study – to establish the protein oxidative modification in blood and periodontium in case of periodontitis combined with hyper- and hypothyroidism in rats. Research Methods. Experimental studies were carry out on 48 mature male, nonliner, white rats, which were divided into the following groups: І – control animals; ІІ – animals with periodontitis; ІІІ – animals with periodontitis combined with hyperthyroidism; IV – animals with periodontitis combined with hypothyroidism. Protein oxidative modification (POM) was determined in blood serum and periodontium homogenate by the method of I.F. Meschyshyn. Results and Discussion. The results of our studies showed that the intensity of the processes of protein oxidative modification was significantly increased in animals of all experimental groups vs control group. The content of aldehyde- and ketone derivatives of the alkali nature changed more pronounced than the content of aldehyde and ketone derivatives of neutral nature. Conclusions. Thus, the experimental periodontitis is accompanied by a marked increase in the intensity of the protein oxidative modifications both in the homogenate of periodontal tissues and in the blood serum. Imbalance of thyroid hormones increases oxidative stress in experimental periodontitis, especially pronounced in hyperthyroidism.

INFLUENCE OF LOW-INTENSITY RED LIGHT ON PROTEIN OXIDATIVE MODIFICATION IN RADIATION-INDUSED DISEASE OF LUNGS IN EXPERIMENT

The aim of the study was to investigate the content of spontaneous protein oxidative modification (POM) in the blood serum and pulmonary tissue of rats after local gamma irradiation of the projection area of the heart and lungs. The content of products of spontaneous POM (aliphatic neutral and basic aldehyde- and ketone-dinitrophenylhydrazones) in the blood serum and pulmonary tissue of 57 rats was studied. The animals were divided into 5 groups: “control group”, where there was gamma irradiation of the heart and lungs area, the collection of samples was done at the next day; “treatment group”, where a single session of low-intensity broadband red light was performed after local exposure to gamma radiation (the collection of samples was done at the next day); “chronic control group”, where there was an exposure to local gamma irradiation of the heart and lungs area, the collection of samples was done at the fourth day; “chronic treatment group” was daily exposed to low-intensity red light for 4 days after local exposure to gamma radiation, the collection of samples was done at the fourth day; “normal level group” was not exposed to either gamma radiation or broadband red light. In the pulmonary tissue and blood serum of control animals, a gradual increase in the content of spontaneous POM products was observed. The most significant difference from the normal level the “chronic control group” had. At the same time, the normalization of the content of POM products under the influence of low-intensity broadband red light in the tissues of the experimental groups was revealed. In the course of the experimental study, the normalization of processes of free radical oxidation of proteins in the lung tissue and in the serum of rats after gamma irradiation and the exposure of the projection region of the heart and lungs to low-intensity broadband red light was shown. Thus, low-intensity red light can be considered as a possible radioprotector.