Comprehensive morphological study of free radical processes in chronic chorioamnionitis on the background of iron deficiency anemia in pregnancy.

OBJECTIVE: Aim: To establish the features of free radical processes in the endotheliocytes of the chorionic plate of the placenta in chronic chorioamnionitis against the background of iron deficiency anemia of pregnant women using both chemiluminescent and histochemical methods of research. PATIENTS AND METHODS: Materials and Methods: 82 placentas from parturients at 37 - 40 weeks of gestation were studied. Including, for comparison, the placenta during physiological pregnancy and the observation of iron deficiency anemia of pregnant women without inflammation of the placenta. The number of observations in specific study groups is given in the tables. To achieve the objective and solve the tasks set in this study, there were carried out the following histochemical, chemiluminescent, morphometric and statistical methods of material processing. RESULTS: Results: In case of chorionamnionitis against the background of anemia in pregnancy, the R/B ratio (R/B - ratio between amino- (blue) and carboxyl (red) groups of proteins)) in the method with bromophenol blue according to Mikel Calvo was 1.56±0.021, indicators of chemiluminescence of nitroperoxides were 133±4.5, relative optical density units of histochemical staining using the method according to A. Yasuma and T. Ichikawa was - 0.224±0.0015. CONCLUSION: Conclusions: With chronic chorioamnionitis, the intensity of the glow of nitroperoxides, the average indicators of the R/B ratio, and the optical density of histochemical staining for free amino groups of proteins are increased compared to placentas of physiological pregnancy and anemia of pregnant women. Comorbid i anemia of pregnant women causes increasing of the intensity of the glow of nitroperoxides, the average values of the R/B ratio, and the optical density of histochemical staining for free amino groups of proteins comparing to placentas with inflammation without anemia. The key factor in the formation of morphological features of chronic chorioamnionitis with comorbid anemia is the intensification of free radical processes, which is reflected by the increase in the concentration of nitroperoxides in the center of inflammation, with the subsequent intensification of the processes of oxidative modification of proteins, which is followed by the increasing activity of the processes of limited proteolysis.

Gender-related Characteristics of Oxidative Stress Severity and Cognitive Impairment in Patients With Schizophrenia.

The gender specificity of the clinical and psychopathological features of schizophrenia is an important factor in this disease. Gender features of neurocognitive deficit in schizophrenia and associated functional and structural disorders of the brain activity are of particular interest to researchers. There are several potential pathogenetic factors of this disease associated with gender, one of which is considered to be oxidative stress. Stress-induced cell death in the prefrontal and anterior frontal regions and reduced brain volume in these regions lead to cognitive and executive decline in patients with schizophrenia. Despite the great interest in the gender factor in schizophrenia pathogenesis, there are currently very few studies on gender differences in the severity of redox imbalance in patients with schizophrenia and their association with neurocognitive deficit. The aim of the study was to reveal the gender specificity of oxidative stress severity in patients with schizophrenia and assess its association with the severity of neurocognitive deficit. The study included 125 patients with schizophrenia and 75 sex- and age-matched healthy volunteers. Sociodemographic and clinical data were studied. Cognitive functions were evaluated using BACS. Blood samples were taken for biochemical studies of the levels of reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) in erythrocyte hemolysate, malonic dialdehyde (MDA), aldehyde-2,4-dinitrophenylhydrazones (ADNPH), ketone-2,4-dinitrophenylhydrazones (KDNPH) in plasma. Levels of oxidative stress markers were assessed by spectrophotometric method. In a sample of patients with schizophrenia, a statistically significantly higher activity of CAT was revealed among women compared to men (T=2.25; p=0.025), however, it was lower than in healthy volunteers. But, at the same time, a higher concentration of protein peroxidation products was found in the peripheral blood of women than in men (ADNPH MCO: T=2.52; p=0.013; KDNPH MCO: Z=-2.26; p=0.017). In the group of healthy volunteers, in contrast to patients with schizophrenia, gender differences in markers of oxidative stress were not found. In women with schizophrenia, single correlations were found between the level of the lipid peroxidation product MDA and the test scores for verbal memory (R=-0.36; p=0.006) and working memory (R=-0.36; p=0.006), antioxidant enzyme activity SOD and motor skills (R=-0.26; p=0.047). In men, on the contrary, multiple correlations of both antioxidants and, mainly, products of lipid and protein peroxidation with cognitive functions assessed using BACS were found. Despite the fact that oxidative stress is more pronounced in women than in men with schizophrenia, associations of redox imbalance with neurocognitive deficit in women is much less pronounced than in men.

[Comparative characteristics of oxidative modification intensity of proteins in infant milk formulas].

Cow's milk is mainly used in the production of infant milk formulas. However, the protein composition of cow's milk differs significantly from the proteome of breast milk. In addition, various technological factors significantly affect the properties and structure of proteins, susceptibility to oxidative processes. This article uses a method of complex evaluation of the products of oxidative modification of proteins to characterize the total level of carbonyl compounds with the analysis of the ratio of aldehyde-dinitrophenylhydrazones (ADNPH) and ketone-dinitrophenylhydrazones (CDNPH), which increases the possibility of determining the severity of protein damage. The purpose of the study was to compare the level of indicators of oxidative modification of proteins in adapted milk infant formulas. Material and methods. The research objects were 4 dry adapted milk infant formulas, as well as ultra-pasteurized cow's milk. The intensity of oxidative modification of milk proteins was determined spectrophotometrically by the reaction of carbonyl compounds with dinitrophenylhydrazine (DNFG). Results. With spontaneous oxidation in infant formula, the total area of carbonyl derivatives of proteins (Sомб) was increased compared to the indicators of ultra-pasteurized cow's milk. The greatest change occurred in relation to ADNFG indicators (SАДНФГ), the level of which increased in 3 formulas by 48.6-59.4%. The content of ketone derivatives (SКДНФГ) did not differ significantly in the studied mixtures from milk indicators. The level of carbonyl derivatives of proteins in milk infant formulas was even more elevated during the induction of oxidation by the addition of iron ions and hydrogen peroxide to the incubation medium. The content of both total and various fractions of carbonyl compounds exceeded the corresponding indicators of cow's milk by 2.0-2.6 times. Conclusion. The results of the study indicate a greater susceptibility to oxidative damage of proteins in milk infant formulas, compared with cow's milk.

PECULIARITIES OF DYNAMICS OF INDICATORS OF PROTEINS OXIDATIVE MODIFICATION AND MATRIX METALLOPROTEINASE-9 ACTIVITY IN PATIENTS WITH PARANOID SCHIZOPHRENIA DEPENDING ON THE DISEASE DURATION.

OBJECTIVE: The aim: The objective of the research was to study the indicators of oxidative modification of proteins (OMP) and the activity of matrix metalloproteinase-9 (MMP-9) in patients with paranoid schizophrenia depending on the disease duration. PATIENTS AND METHODS: Materials and methods: 320 patients were included in the examination. 20 patients were with "Primary psychotic episode" (Comparison Group) and 300 patients were diagnosed with "Paranoid schizophrenia" (Experimental Group): 60 of them have suffered from this disease for a duration from 3 to 5 years (Subgroup I ); 60 patients have suffered for a period from 6 to 10 years (Subgroup II); 60 individuals - from 11 to 15 years (Subgroup III); 60 patients have suffered for a duration from 16 to 20 years (Subgroup IV); 60 patients - from 21 years and longer (Subgroup V). RESULTS: Results: The presented data showed that the levels of OMP indicators in Subgroup I constituted 0.826±0.046 conventional units at a wavelength of 356 nm; 0.864±0.051 conventional units at a wavelength of 370 nm; 0.444±0.019 conventional units at a wavelength of 430 nm; 0.176±0.007 conventional units at a wavelength of 530 nm, which is 1.99; 1.6; 1.13 and 1.43 times higher than in the Comparison Group. The content of OMP products was higher by 2.24; 1.74; 1.17, and 1.43 times in Subgroup II, respectively, by 2.4; 1.80; 1.36 and 1.46 times in Subgroup III, respectively; by 2.5; 1.9; 1.4; 1.6 times in Subgroup IV, respectively; by 2.5; 2.02; 1.54; 1.7 times in Subgroup V, respectively. The conducted correlation analysis indicated a direct correlation between OMP indicators and the disease duration. The concentration of MMP-9 in the patients of the Comparison Group was equal to 892.84±87.80 pg/ml, which was 11.2% less compared to the Experimental Subgroup I, where this indicator was 992.84±67.50 pg/ml. MMP-9 constituted 1092.53±47.20 pg/ml on average in the patients of Subgroup II, which was 22.36% higher than in the Comparison Group. This indicator was 1702.84±37.60 pg/ml in Subgroup III, which was 90.7% higher than in the Comparison Group. It constituted 1492.84±47.29 pg/ml in Subgroup IV, which was 67.2% higher than in the Comparison Group; and 2037.21±57.80 pg/ ml in Subgroup V, which was more than two times higher than in the Comparison Group (p<0.05). The conducted correlation analysis showed a direct relation between MMP-9 expression and the increase in OMP indicators. This relation was more significant between MMP-9 and OMP products of a neutral nature. The correlation strength between MMP-9 and OMP products of a basic nature was somewhat less significant. CONCLUSION: Conclusions: According to the results of the conducted analysis, the examined patients had the signs of decompensation of reactive-adaptive biomolecular mechanisms which activated radical reactions with the subsequent accumulation of oxidation products.

Influence of Rectal Ozone Application on the Intensity of Free Radical Destruction of Lipids and Intestinal Proteins in the Dynamics of Experimental Colitis.

We studied clinical status, content of products of LPO, and oxidative modification of proteins (OMP) in the lesion focus of the intestine in experimental colitis under conditions of rectal administration of ozone. Experimental colitis was simulated by two-stage administration of oxazolone; rectal insufflation of ozone in the ozone-oxygen mixture was performed daily. The disease activity index (DAI), the content of calprotectin in the feces, and LPO and OMP products in the intestinal homogenate were assessed. On days 2, 4, and 6 of the pathological process, DAI, concentration of calprotectin in feces, content of primary, secondary, and end-products of LPO in the heptane and isopropanol phases, and content of primary and secondary OMP products progressively increases. Under conditions of ozone application, DAI, concentration of calprotectin in feces, the levels of heptane- and isopropanol-soluble primary, secondary, and end-products of LPO, and the level of primary and secondary products of OMP decreased on days 4 and 6; the level of isopropanol-soluble primary, secondary, end-products of LPO increased on day 2 of experimental colitis. The severity of clinical manifestations weakens as the content of LPO and OMP products in the colon decreases on days 4 and 6 of observation.

Association of antioxidant deficiency and the level of products of protein and lipid peroxidation in patients with the first episode of schizophrenia.

Schizophrenia is considered a multifactorial disease, where one of the pathogenetic links is oxidative stress; however, the results of studies are often contradictory, largely due to significant heterogeneity among study methods. The present study was undertaken to compare the levels of oxidative stress markers in the peripheral blood of patients with a first episode of schizophrenia (FES) and in healthy volunteers (HV). The study included 50 patients with FES and 37 HV. Blood samples were collected for spectrophotometric assessment of reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), aldehyde-2,4-dinitrophenylhydrazone (ADNPH), and ketone-2,4-dinitrophenylhydrazone (KDNPH) in blood plasma. Results showed that in patients with FES compared with HV, a significant decrease in CAT activity and an increase in oxidative modification of proteins (OMP) were found. In both groups, a significant increase in the level of MDA with age was revealed. In patients, the GSH plasma level was inversely proportional to the ADNPH level, and SOD activity was directly proportional to the KDNPH level. In volunteers there was no such correlation; however, there was a direct correlation between CAT activity and the levels of OMP and MDA. In both groups, a moderate direct correlation between peroxidation products was observed. The results confirm that a redox imbalance (a deficiency of antioxidants, in particular CAT, and excess OMP) may be a pathogenetic link in schizophrenia, which is manifested already at an early stage of the disease.

Effect of Vitamin D3 in Composition of Original Rectal Suppositories on Parameter of Protein Oxidative Modification in Large Intestine in Experimental Ulcerative Colitis.

The effect of vitamin D3 in the composition of original rectal suppositories on the content of products of oxidative modification of proteins in mucous membrane of the large intestine was studied in rats with experimental ulcerative colitis provoked by a two-stage administration of 3% oxazolone. The rectal suppositories with vitamin D3 (1500 IU) were administered every 12 h during 5 days. Condition of the rats was assessed according to disease activity index (DAI), while the content of oxidative modification products of proteins in the homogenate of the mucous membrane was assayed with extraction-spectrophotometric method in the lesion focus of large intestine. DAI increased during entire observation period of ulcerative colitis, which correlated with the level of products of spontaneous and induced oxidative modification of proteins in mucous membrane of the colon. The study examined the pharmaceutical and technological features of novel rectal suppositories of original composition weighing 300 mg, which are based on polyethylene glycol supplemented with aqueous solution of vitamin D3 (10%). The use of rectal suppositories with vitamin D3 reduced DAI and inhibited the oxidative modification of proteins.

Structural and Morphological Changes in the Liver Due to Intestinal Endotoxins.

BACKGROUND: Under normal physiological conditions, endotoxin (ET) released during self-renewal of the colibacillus pool is an obligate stimulus for the formation of the immune system and homeostasis of the body. Violation of the barrier function of the intestinal wall and the mechanisms of neutralization of endotoxin lead to systemic endotoxemia of intestinal origin. Its development is facilitated by stress, intoxication, a decrease in nonspecific resistance of the body, as well as damage to the intestinal mucosa and dysbiosis, where the mucous membrane is more vulnerable and permeable to endotoxin. PURPOSE OF THE RESEARCH: The aim of this study is to compare and assess the severity and nature of hepatocyte damage from endotoxin exposure and the degree of manifestation of stress due to oxidation, to determine the characteristics of structural changes in hepatocytes and to assess the oxidation stress during endotoxin intoxication in the experiment with biochemical markers. MATERIALS AND METHODS: The experiments were conducted on 40 non-linear rats, divided into two groups of 20 animals. Group 1 animals received intraperitoneal injections of ET of Escherichia coli drug (Sigma USA K-235) for seven days at a rate of 0.1 mg/kg of the body weight. Animals of the second group served as the control group. Character and stage of liver damage were studied using morphological methods, including electron and light microscopy. In studying oxidizing stress, biochemical methods were used to define the changes, such as conjugated dienes and dienketones, spontaneous oxidizing modification of proteins. RESULTS AND CONCLUSION: 1. The severity and depth of morphological changes in the liver during endotoxin intoxication were correlated with the dynamics of the content of lipid oxidation products (CD and DK, MDA) and proteins. There was a tendency for a more significant increase in the oxidative modification of proteins in serum. This confirms the data on the primary damage of proteins by free radicals. 2. When exposed to intestinal microflora endotoxin, pronounced dyscirculatory changes, fatty and hydropic degeneration of hepatocytes with signs of toxic damage to their nuclei were determined, but at the same time, the increased hyperplastic activity of sinusoidal cells remained associated with the effects of endotoxin. These changes are associated with both the direct toxic effect of endotoxin, and the effects of oxidative stress, in which endotoxin is a potent inducer.

Association between Advanced Glycation End Products, Soluble RAGE Receptor, and Endothelium Dysfunction, Evaluated by Circulating Endothelial Cells and Endothelial Progenitor Cells in Patients with Mild and Resistant Hypertension.

The aim of the present study was to evaluate advanced glycation end products (AGEs) and soluble form of receptor RAGE (sRAGE) concentrations as well as the AGEs/sRAGE ratio in mild (MH) and resistant (RH) hypertensive patients in comparison with normotensive individuals. We also evaluated the association between AGEs, sRAGE as well as AGEs/sRAGE ratio and circulating endothelial cells (CECs) and circulating endothelial progenitor cells (CEPCs). The MH group consisted of 30 patients, whereas 30 patients were classified for the RH group. The control group (C) included 25 normotensive volunteers. AGEs and sRAGE were measured using enzyme-linked-immunosorbent assay (ELISA). The multicolor flow cytometry was used for analysis of CECs and CEPCs. Significantly higher levels of AGEs in RH cohort were observed as compared to C cohort. Furthermore, significantly lower sRAGE levels as well as a higher AGEs/sRAGE ratio were observed between MH and RH cohorts. Significant correlations were found in the MH cohort for sRAGE and CECs, and CEPCs. The elevation of AGEs levels suggests that oxidative modification of proteins occurs in hypertension pathogenesis. The decrease in sRAGE levels and elevation of the AGEs/sRAGE ratio in MH and RH groups may suggest that hypertensive patients are less protected against the side effects of AGEs as a consequence of an insufficient competitive role of sRAGE against the AGEs-RAGE axis. Finally, it may be concluded that the level of AGEs may be an independent predictor of the condition and function of the endothelium. Furthermore, sRAGE may be classified as a potential biomarker of inflammation and endothelium dysfunction.

Effects of flavonol quercetin on activity of lipid peroxide oxidation in experimental bacterial-immune periodontitis.

The article presents the results of lipid peroxidation activity in blood serum, which were assessed on the base changes in the level of oxidative modification of proteins, TBA-active products, and total amount of nitrogen (II) oxide metabolites (NO2 - + NO3 -). Indices were determined on the 14th day of development in experimental periodontitis both without correction, and in condition treatment with a water-soluble flavonol quercetin (corvitin). The treated animals take intramuscular injections of corvitin in a dose 100 mg/kg weight for 7 days. For further testing, blood serum was selected. The results were statistically analyzed using parametric and non-parametric indices. In this investigation, characterized dynamics of changes in the indices of oxidative processes activity were revealed both during the period of formation and the course of periodontitis. In particular, it was found that, for the period of the most expressed inflammatory reaction in the periodontal complex, intensive increase in the level of reactive forms of oxygen. The treatment with flavonol quercetin for 7 days resulted in stabilization of free radical processes and the suppression of the inflammatory reaction.

Influence of Low-Intensive Red Light on the Myocardium in Experimental Asphyxia.

We studied the effects of low-intensity broadband red light on electrical activity of the heart and oxidative modification of proteins in the myocardium of rats after asphyxia. It was shown that low-intensity red light reduced the content of oxidatively modified proteins in rat heart after oxidative stress caused by asphyxia. Exposure to low-intensity red light normalized ECG parameters in rats after asphyxia.

[The characteristics of redox-status of peripheral part of erythron under various localizations of neoplasm of organs of female reproductive sphere.]

The study was carried out to evaluate characteristics of redox-status in erythrocytes of peripheral blood of patients with ovarian cancer, uterine body cancer and cervix cancer using evaluation of products of peroxide oxidation of lipids: diene conjugates, ketodienes, Schiff's bases by Volchegorskii, malonic di-aldehyde by L.I. Andreieva, content of products of oxidizing modification of proteins by E.E. Dubinina; activity of anti-oxidant enzymes: superoxiddismutase by Nishikimi, catalase, glutathione-S'-transferase, glutathionereductase and level of reduced glutathione by A.I. Karpischenko. The evaluation of architectonics of erythrocytes and rigidity of membrane was implemented using a scanning probing microscope SolverPro (NT-MDT, Zelenograd, Russia). The statistical processing of obtained data was implemented using non-parametric Mann-Whitney U-test. The increasing of products of initial stages of peroxide oxidation of lipids - diene conjugates and malonic di-aldehyde and decreasing of level of interim (ketodienes) and tertiary ( Schiff's bases) products of peroxide oxidation of lipids have been established. Against this background a multi-directional alteration of superoxiddismutase and decreasing of catalase occurred. Simultaneously, increasing of activity of glutathione-transferase and level of reduced glutathione under all analyzed localizations of neoplasm were noted. The level of products of oxidizing modification of proteins depended on localization of primary tumor and it was minimal in case of cervix cancer. Also such an alteration of cyto-architectonics of erythrocytes was established as development of reversible deformed echinocytes in patients in case of ovarian cancer and irreversible deformed spherocytes in patients with cervix cancer and uterine body cancer and also abrupt increasing of rigidity of membrane of erythrocytes. The obtained data permits to surmise as a capacity of para-neoplastic processes development of oxidative and carbonyl stress in erythrocytes of peripheral blood of patients with ovarian cancer, cervix cancer and uterine body cancer at extensive stage of disease. The mentioned stress is accompanied by formation of echinocytes and spherocytes and significant increasing of rigidity of membrane. However, intensity of these processes is determined by localization of primary neoplasm and it can be applied for characteristic of biological portrait of tumor in case of consideration of schemes of anti-oxidant therapy.

Oxidative modification of proteins in the process of experimental periodontitis development.

The article reveals patterns of changes in the parameters of oxidative modification of proteins for different periods of the inflammatory process in periodontal tissues during periodontitis. Biochemical researches of phenylhydrazones, aldehyde-, and ketone derivatives of neutral and basic proteins were determined in the blood of white rats on the 7th, 14th, and 30th days of the experimental periodontitis development, as well as in intact animals. The method for determination of the oxidative modification of proteins was based on the interaction of oxidized amino acid residues with 2,4-dinitrophenylhydrazine to form 2,4-dinitrophenylhydrazones. At the early stage of experimental periodontitis development, i.e., on the 7th day, an increase in the products of oxidative modification of proteins of basic and neutral nature was observed in the blood serum, but later, on the 14th day, this index changed in opposite direction, i.e., it began to decrease; however, it was higher relative to the intact group of animals. The obtained factual data evidence that under the conditions of experimental periodontitis formation, there is an intensive increase in the level of oxidative modification of proteins especially in the early period of the inflammatory process development.