Reactive Carbonyl Derivatives Research Articles

Evaluation of clinical features of bronchial asthma of varying severity and the level of modified proteins

The article presents the results of a study of 62 patients with bronchial asthma (ВА), which were divided into 2 groups: the first group included patients (n=34) with moderate severity of asthma, the second group included patients (n=28) with severe asthma. The worsening of the course of bronchial asthma in the examined patients is due to the duration of the disease, the presence of harmful risk factors, among which adverse living conditions with the development of sensitization to them are of the greatest importance. In severe bronchial asthma, an uncontrolled course of the disease prevails with a 1.8-fold decrease in asthma-control test values (p<0.05). With an increase in the severity of the inflammatory process in the airways, the level of AOРP in the blood plasma increases. In patients with bronchial asthma, the following features of oxidative metabolism were established: in the blood plasma and erythrocytes of patients with bronchial asthma of varying severity, a tendency to an increase in reactive carbonyl derivatives of proteins, malondialdehyde, AOPP compared with the control was observed, which indicates an increase in oxidative metabolism with the development of pathology.

Clinical features and evaluation of modified proteins in bronchial asthma of various severity

The article presents the results of a study of 62 patients with bronchial asthma (ВА), which were divided into 2 groups: the first group included patients (n = 34) with moderate severity of asthma, the second group included patients (n = 28) with severe asthma. The worsening of the course of bronchial asthma in the examined patients is due to the duration of the disease, the presence of harmful risk factors, among which adverse living conditions with the development of sensitization to them are of the greatest importance. In severe bronchial asthma, an uncontrolled course of the disease prevails with a 1.8-fold decrease in asthma-control test values (p<0.05). With an increase in the severity of the inflammatory process in the airways, the level of AOРP in the blood plasma increases. In patients with bronchial asthma, the following features of oxidative metabolism were established: in the blood plasma and erythrocytes of patients with bronchial asthma of varying severity, a tendency to an increase in reactive carbonyl derivatives of proteins, malondialdehyde, AOPP compared with the control was observed, which indicates an increase in oxidative metabolism with the development of pathology.

Uticaj vitamina B6 na markere oksidativnog stresa, kardiometaboličke i histološke markere u uslovima monokrotalinom indukovane srčane slabosti kod Wistar albino pacova

Introduction/Aim: Heart failure (HF) induced by monocrotaline (MCT) is common in the pulmonary arterial vessels remodeling mechanisms with increased pulmonary resistance and oxidative stress markers. The purpose of this study was to validate the hypothesis that the treatment with vitamin B6 could affect HF by modulating cardiometabolic and oxidative stress biomarkers, and the structure of the rat heart. Material and Methods: Male Wistar albino rats were divided into 3 groups: blank solution-exposed control (C physiological saline 1ml/kg 28 days ip., n=8), B6 (vitamin B6 7mg/kg/day 28 days ip., n=8), and MCT+B6 (MCT 50mg/ kg once ip. plus vitamin B6 7mg/kg/day 28 days ip., n=8). Results: The four-week vitamin B6 treatment significantly affected certain biochemical parameters. The activity of key antioxidant enzymes, superoxide dismutase (SOD) and glutathione peroxidase (GPX) did not change, whereas the total glutathione (GSH) was significantly decreased in the MCT+B6 group. This was followed by a slightly decreased level of the total glutathionylation observed in the MCT+B6 group. The parameters of protein oxidative damage (reactive carbonyl derivates, thiol groups and nitrotyrosine) did not significantly change in the MCT+B6 group. An increasing trend in RV and LV wall thickness was observed in the MCT+B6 compared to the C group, as well as in Ki67 and PCNA positivity. Conclusion: The four-week treatment with vitamin B6 significantly affected certain biomarkers. The activity of SOD and nitrotyrosine content did not change, while GPX activity, total glutathione and total glutathionylation level were decreased in the MCT+B6 group. We observed an increase in RV and LV wall thickness in the MCT+B6 group compared to the C group, as well as in Ki67 and PCNA positivity.

Активність цитозольних ензимів катаболізму ендогенних альдегідів у печінці щурів за умов різної забезпеченості раціону нутрієнтами

The research was conducted to study the activity of aldehyde dehydrogenase (EC 1.2.1.3) and aldehyde reductase (EC 1.1.1.21), the levels of TBA reactive substances and protein carbonyl derivates in the cytosolic fraction of rat liver under the conditions of different dietary sucrose and protein content. The animals were distributed into the 4 experimental groups: group I — animals receiving full-value semi-synthetic feed (control group); group II — animals on a low-protein diet (LPD); III group — animals on a high-sucrose diet (HS); IV group — animals on a low-protein and high-sucrose diet (LPD/HS). It was found that in animals under conditions of dietary protein deficiency, there was a two-fold increase in the levels of TBA reactive substances and protein carbonyl derivates in the liver cytosolic fraction against the absence of changes in the aldehyde reductase and aldehyde dehydrogenase activity. At the same time, in animals on a high-sucrose diet, there was a significant accumulation of the TBA reactive substances and carbonyl derivatives in the liver cytosolic fraction along with a 2–2.5-fold increase in both aldehyde reductase and aldehyde dehydrogenase activity. The maximum accumulation of the products of oxidative damage to proteins and lipids along with the insufficient activation of the enzymes ensuring their catabolism can be considered as one of the possible mechanisms of liver cell damage under conditions of the low-protein/high-sucrose diet. The obtained results open new prospects for future studies of the mechanisms of endogenous aldehydes detoxification and further development of a strategy for the correction of metabolic liver disorders under the conditions of nutrient imbalance.

The Reactive Carbonyl Derivatives of Proteins, Methylglyoxal, and Malondialdehyde in Blood of Women with Breast Cancer

BACKGROUND: Every year 1.5 million women in the world are diagnosed with breast cancer (BC). In 2018, more than 260,000 new cases of cancer and more than 40,000 deaths due to this disease were detected. At the same time, in Kazakhstan, an intensive indicator of the incidences of BC in 2018 amounted to 25.3% per population of 100 thousand people (2017–24.5%) with a growth rate of 3.1%, which in absolute numbers are 4,648 new cases per year. In terms of mortality, BC ranks third after lung and stomach cancer (6.8%). AIM: This necessitates a detailed study of the molecular mechanisms that underlie the development and progression of BC. One of the mechanisms of carcinogenesis is oxidative stress (OS). An increase in malondialdehyde (MDA) levels was detected in the early stages of cancer. It was suggested that MDA, due to its high cytotoxicity, acts as a promoter of the tumor and cocarcinogen agent. METHODS: Therefore, violation of the parameters of OS in BC is in no doubt. However, according to the literature data analysis, these results are ambiguous and contradictory. There are no studies on a comprehensive assessment of the oxidative destruction of lipids, proteins, and nucleic acids in BC. CONCLUSION: The nature and direction of changes in various components of OS in patients with BC have not been adequately studied, which is necessary for a correct assessment of the involvement of OS in the mechanism of the pathological process and determination of a sensitive marker of the risk of BC or its progression.

Molecular patterns of oxidative stress in drug-induced nephropathy

Introduction: Drug induced kidney disorder is a frequent adverse event which contributes to morbidity and even incapacitation. The discovery and development of novel biomarkers and local (renal) response mechanisms, are needed for effective prevention of drug-induced nephrotoxicity. Objectives: The main purpose of our study was to investigate the oxidative modifications of proteins in blood plasma and erythrocytes of patients with drug-induced nephropathies. Patients and Methods: Around105 patients were divided into two groups: first group was represented by patients with psychotropic drug-induced nephropathy; the second one consisted of patients received nonsteroidal anti-inflammatory drugs (NSAIDs). Advanced oxidation protein products (AOPPs) were measured by Witko-Sarsat method. Protein reactive carbonyl derivatives (PRCD) were assayed in blood plasma and erythrocytes by the Levine method. Neutrophil gelatinaseassociated lipocalin (NGAL) was determined with the use of a commercially available ELISA kit. Results: Carbonyl derivatives are significantly higher in red blood cells of the 1st and 2nd group patients compared to the control subjects. AOPP statistically increased both in patients with various types of drug nephropathy and in patients with chronic kidney disease (CKD) compared with the control group. The NGAL was significantly higher in all groups compared to the control subjects. Conclusion: The patients with drug-induced nephropathy have increased level of oxidative stress products and response NGAL reaction. The mechanisms that lead to the development of oxidative stress and the production of modified proteins are different in patients treated with different drugs. Establishing patterns of cell-molecular interaction permit the drug-induced nephropathy to be timely diagnosed and therapeutic programs to be optimized.

The Level of Reactive Carbonyl Derivatives of Proteins, Malondialdehyde, and Catalase Activity in the Brain of Rats after Therapy Following Chronic Unpredictable Moderate Stress

BACKGROUND: Understanding the mechanisms of the behavioral disorders’ emergence under the influence of chronic stress is the most important aspect of the subsequent development of a strategy for its therapy and prevention. Changes in the oxidative metabolism processes can be decisive in the development of the pathogenetic cascade in the brain. Information about these processes can be obtained by studying protein carbonylation, lipid peroxidation, and catalase activity (CA). The complexity of the therapeutic impact in various behavioral disorders implies the search for new pharmacological substances and the study of the previously known drugs’ effects based on the available scientific data.
 AIM: The aim of the study was to study the reactive carbonyl derivatives of proteins (RCDP), malondialdehyde (MDA), and CA in the brain of rats after therapy following chronic unpredictable moderate stress (CUMS).
 METHODS: Forty male outbred rats weighing 450–500 g were used in this study. For 21 days, all animals were exposed to the diverse stress factors for developing the CUMS. The animals were divided into four groups of 10 rats, each using randomized selection. The rats of one group were euthanized by decapitation with subsequent brain harvesting (Group 4). Remaining three groups of rats were treated with placebo (Group 1), harmine hydrochloride (Group 2), and amitriptyline (Group 3) for 21 days. Upon completion of therapy, all rats were also euthanized by decapitation with subsequent brain harvesting. The levels of RCDP, MDA, and CA were studied in their brain, and after that, we compared the multiple studied indicators in four groups.
 RESULTS: The results of the rat brain examinations in four groups showed that RCDP level in Group 2 was significantly lower than in Group 4 (p = 0.000). Similarly, in Group 1, it was lower than in Group 4 (p = 0.021), plus, it did not differ statistically from the harmine hydrochloride group (p = 1,000). Indicators of Groups 3 and 4 did not have any significant differences in RCDP level, too, (p = 0.799); however, the RCDP level in Group 2 was significantly lower than in Group 3 (p = 0.040). MDA indicators did not show significant differences; however, a tendency for lower values was revealed in Group 1 (p = 0.233) and Group 2 (p = 0.151). CA in Group 4 was lower than that in Group 1 (p = 0.000), Group 2 (p = 0.001), and Group 3 (p = 0.003) contemporaneously, while all treatment groups were comparable (p = 1.000).
 CONCLUSION: The result of exposure to chronic stress can be reproduced with the best quality in the CUMS model. The neurobiological foundations of the model make it possible to assess biochemical markers of oxidative metabolism and evaluate the possibilities of pharmacological correction of stress-induced behavioral disorders. To assess the mechanisms of autoregulation of oxidative metabolism, this study included a placebo group (Group 1), the level of RCDP in which was significantly higher in comparison with Group 3 and Group 4 and slightly lower than in Group 2. In this study, harmine hydrochloride demonstrated activity exceeding amitriptyline, particularly limiting the process of protein carbonylation, not noted for amitriptyline. According to the results of the RCDP assessment in the CUMS model, the process of protein carbonylation can be considered to be one of the significant factors in the deactivation of neurotransmitters. The CA levels determined in all groups allowed us to consider this marker as the most sensitive to the effects of stress, which possibly has an inhibitory effect on catalase, as its activity in all groups after therapy was more than two-fold higher than in animals right after CUMS. We can assume that CA plays an important role in starting the processes of autoregulation of oxidative metabolism. The study was carried out as a part of the implementation of the scientific and technical program No. BR05236584 “Development of new herbal preparations and their pharmacological and clinical studies” (O.0820). (2018–2020) in the priority area, “Life and Health Sciences.”

Drug-induced Nephropathy and Molecular Patterns of Oxidative Stress

BACKGROUND: Drug-induced kidney disorder is a frequent adverse event which contributes to morbidity and even incapacitation. Our current knowledge of drug-induced kidney disease is limited due to varying definitions of kidney injury, incomplete assessment of concurrent risk factors, and lack of long term outcome reporting.
 AIM: The discovery and development of novel biomarkers and local (renal) response mechanisms, which can diagnose kidney damage earlier and more accurately, are needed for effective prevention of drug-induced nephrotoxicity.
 METHODS: Forty patients from 22 to 50 years old with drug-induced nephropathy (43 females, 27 males) were included in the study. The determination of oxidative stress modifications as advanced oxidation protein products (AOPPs), protein reactive carbonyl derivatives (PRCD), methylglyoxal, and catalase (CAT) level were assayed. AOPP was rated in blood plasma through the Witko-Sarsat (1996) method, PRCD by Levine (2000) method, methylglyoxal through the Racker (1963) method, CAT was evaluated with Koroljuk (1988) and Aebi (1984) methods. Conditionally, 30 healthy-matched persons were included in the control group.
 RESULTS: In psychotropic and nonsteroidal anti-inflammatory drugs (NSAID) drug-induced nephropathy patients, there was a tendency to increase the concentration of AOPP in blood plasma, PRCD, and methylglyoxal in erythrocytes relative to the upper limits of normal ranges. Moreover, there was a significant decrease of the CAT level in blood plasma relative to the upper limits of normal ranges.
 CONCLUSION: Thus, the results of the present study permit us to draw the following conclusions. The patients with psychotropic and NSAIDs drug-induced nephropathy have increased level of oxidative stress products and increased neutrophil gelatinase-associated lipocalin level even in normal eGFR. The mechanisms that lead to the development of oxidative stress and the production of modified proteins are different in patients treated with different drugs.

The Level of Reactive Carbonyl Derivatives of Proteins, Methylglyoxal, and Malondialdehyde in Rats Experiencing Chronic Unpredictable Moderate Stress

BACKGROUND: Prolonged exposure to stress is known to change homeostasis and can lead to the development of psychopathological disorders, including depression. Exposure to stress factors contributes to the emergence of reactive oxygen species in the brain. The data characterizing changes of the oxidative metabolism under conditions of chronic stress are insufficient, which determine the relevance of the research in this direction. The determination of reactive carbonyl derivatives of proteins (RCDP), methylglyoxal (MG), and malondialdehyde (MDA) levels can be considered an informative method for studying the dynamics of oxidative metabolism.
 AIM: The aim of the study was to study the RCDP, MG, and MDA in red blood cells and blood plasma in rats experiencing chronic unpredictable moderate stress (CUMS).
 METHODS: A total of 20 male outbred rats weighing 450–500 g were used in the study. The animals were divided into two groups of ten rats each using randomized selection. The experimental group of animals was exposed to the impact of diverse stress factors, according to the CUMS model for 21 days. The control group of animals remained in standard conditions and did not undergo CUMS model. Blood sampling was performed twice for all rats the day before the experiment started and on 21st day to determine the level of RCDP, MG, and MDA in their erythrocytes and blood plasma.
 RESULTS: The study revealed a significant decrease of RCDP (p = 0.007) and MDA levels in erythrocytes (p = 0.013), and MDA level in blood plasma (p = 0.005) of rats of the main experimental group. The control group showed a significant increase of RCDP level in plasma (p = 0.008), a two-fold increase of MDA in erythrocytes (p = 0.05) with a decrease of MDA in plasma (p = 0.011). A tendency for MG increase was noted in erythrocytes of rats in both the main group and the control group.
 CONCLUSION: The early stages of CUMS were accompanied by the violation of oxidative metabolism with the emergence and accumulation of RCDP and MDA in the blood of animals. The decreased emergence of RCDP and MDA in the later stages can be associated with an excess of oxidized substrates in the blood of animals of the experimental group. The revealed trend toward MG growth in animals of both groups suggests its increased synthesis in erythrocytes, which, in our opinion, should be considered a negative factor affecting the deformability of red cells. In turn, a violation of the deformability of red blood cells leads to a violation of microcapillary blood flow, which contributes to the development of hypoxic damage in different tissues. Thus, the obtained data confirm the different directions of changes in RCDP and MDA levels in the blood of animals experiencing CUMS or acute stress. Our results prove the need for further research on changes in oxidative metabolism in chronic stress disorders.

The Oxidized proteins and activity ofthe Cl-/HCО3-exchangerin erythrocytes of patients withacute alcohol intoxication.

The purpose of this research was to study the morphological properties and the products of oxidative protein modification in erythrocytes of patients with acute alcohol intoxication. Two groups of subjects were analyzed. The first one included 39 patients with acute alcohol intoxication. The second group consisted of 14 healthy subjects. In erythrocytes the activity of Cl-/HCО3- exchanger, the reactive protein carbonyl derivatives and membrane-bound hemoglobin concentration were measured. Our results demonstrated strong alteration of the Cl-/HCО3- exchanger activity in erythrocytes of patients with acute alcohol intoxication. A delay in the beginning of hemolysis during incubation of erythrocytes in the ammonium medium was observed. The concentration of protein carbonyls in erythrocytes of patients significantly increased in comparison to the control ones. A decrease in the membrane-bounded hemoglobin was observed as well. These findings indicate that ethanol toxicity is manifested by alteration of oxidized protein concentration and Cl-/HCО3- exchanger activity in erythrocytes. It is hypothesized that oxidized proteins are implicated in modulation of the erythrocyte cell volume regulation.

P-27 - Oxidazed proteins in blood of patients with nephropathies of various origins

The main purpose of our study was to investigate the oxidative modifications of proteins in blood of patients with nephropathies of various origins. There were 80 patients divided into 4 groups depending on the type of nephropathy. 1st group was represented by patients with acute alcoholic nephropathy; the 2nd group - by patients with nephropathy caused by acute poisoning with alcohol surrogates; in the 3th group pregnant women with gestosis were included; the 4th group – was represented by patients with acute medication nephropathy (acute drug poisoning). The control group consisted of 15 healthy subjects. The reactive protein carbonyl derivatives were detected in plasma and erythrocytes (Levine et.al, 1990), advanced oxidation protein products (AOPP) were detected in serum (Witko-Sarsat et al., 1996). In blood plasma of patients of 1–4 groups the concentration of reactive protein carbonyl derivatives was lower in comparison with control ones (p

The changes of oxidative stress and endothelial function biomarkers after 6 weeks of aerobic physical training in patients with stable ischemic coronary disease

Introduction: Exercise-based cardiac rehabilitation improves endothelialBackground/Aim. Exercise-based cardiac rehabilitation improves endothelial function, reduces cardiac mortality and anginal symptoms in patients with established cardiovascular disease. We evaluated the changes of oxidative stress and endothelial function biomarkers after 6 weeks of aerobic physical training in patients with stable ischemic coronary disease (IHD) participating in a cardiovascular rehabilitation exercise program. Methods. Thirty-five patients with stable IHD engaged in cardiovascular rehabilitation program with the regular aerobic physical activity during 6 weeks were consecutively included together with 37 control, age and sex-matched, IHD patients with a sedentary lifestyle. Clinical data about anthropometric and cardiovascular parameters and laboratory data: serum cell adhesion molecules intracellular (sICAM-1) and vascular (sVCAM-1), reactive carbonyl derivatives (RCD), lipid peroxidation products malondialdehyde (MDA) and nitric oxide (NO) concentration were determined at the beginning and after 6 weeks of aerobic training (45 minutes of continuous exercise up to 80% of maximal heart rate, 3 times a week). Results. The baseline characteristics of examined groups were similar according to age, gender, and cardiovascular risk profiles. The regular aerobic physical activity induced significant reduction of body mass index, blood pressure, heart rate, triglycerides, RCD (1.27 ? 0.48 ?mol/g proteins vs. 1.04 ? 0.22 ?mol/g proteins), sVCAM-1 [100.4, interquartile range (IQR)(78.4?118.3) ng/mL vs. 80.0 IQR(68.5?97.2 ng/mL)] and increasing of NO (64.72 ? 16.06 nmol/mg proteins vs. 74.38 ? 18.57 nmol/mg proteins) and HDL cholesterol (p < 0.05), which was not seen in sedentary control RCD (1.16 ? 0.25 interquartile range vs. 1.12 ? 0.14 interquartile range), sVCAM-1 [92.2 IQR (73.6?106.8 ng/mL) vs. 91.3 IQR (73.0?105.3 ng/mL) and NO (68.5 ? 17.9 nmol/mg vs. 65.7 ? 19.6). The values of sICAM-1 were lower in exercise training group baseline without significant changes during observation [80.74 IQR (54.92?97.3) vs. 80.36 IQR (68.1?95.3)] compared to the control

Evaluation of oxidative stress markers in the heart and liver of rainbow trout (Oncorhynchus mykiss walbaum) exposed to the formalin.

The aim of this study was to examine change in lipid and protein oxidation biomarkers, transamination enzymes and lactate dehydrogenase activities, lactate and pyruvate levels in liver and heart tissue of rainbow trout (Oncorhynchus mykiss Walbaum) that was exposed to formalin baths. Increase of 2-thiobarbituric acid reactive substances and carbonyl derivatives of protein oxidative destruction was noticed only in cardiac tissue of formalin-exposed fish. Activity of lactate dehydrogenase and lactate level in the cardiac tissue were elevated, indicating active glycolysis. Effects of formalin disinfection were different in both tissues. Aldehydic and ketonic derivatives of oxidatively modified proteins in liver were consistently reduced upon exposure to the formalin. In support of this, decrease in alanine and aspartate aminotransferases was noticed. Formalin disinfection of rainbow trout results in metabolic plasticity, predominantly in liver with decreased levels of oxidative stress biomarkers and aminotransferases activity. Formalin-induced oxidative stress in the cardiac tissue was more considerable.

Oxidative metabolism of neutrophils in patients with community-acquired pneumonia

Abstract At the present time, available views show our limited knowledge of the peculiarities of the functional status of neutrophils and their metabolism in patients with community-acquired pneumonia (CAP). The studying of changes of metabolic status of neutrophils can broaden our views about pneumonia pathogenesis and define datum points of therapeutic effect. Purpose of our research: to define oxidative stress activity and the level of oxidative modification of proteins of neutrophils in CAP patients. Materials and methods: neutrophils obtained from 23 patients with community-acquired pneumonia. Control group consisting of 19 healthy volunteers. The reactive carbonyl derivatives of proteins and advanced oxidation protein products were defined so as to assess the oxidative damage of proteins. The malondialdehyde and nitrite ions were assessed as being indicators of the oxidative stress. The neutrophils of CAP patients with moderate severity were characterized by a tendency of evidencing decreasing content of advanced oxidation protein products, along with the statistically important enhanced levels of carbonyl derivatives and nitrite ions, while their malondialdehyde status practically leveled off with the control and had only an insignificant trend towards growth. We have demonstrated the accumulation of carbonyl derivatives and nitrite ions in the peripheral neutrophils of CAP patients. These results give evidence of an oxidative misbalance in the cells which contributes to the aggravation of the disease.

Characteristic of the Oxidative Stress in Blood of Patients in Dependence of Community-Acquired Pneumonia Severity.

BACKGROUND:At the present time the alternation of the oxidative metabolism is considered as one of the leading pathogenic mechanisms in the development and progression of community-acquired pneumonia (CAP). However the nature and direction of the oxidative protein changes in CAP patient’s blood had been almost unexplored.AIM:To define oxidative and modified proteins in erythrocytes and blood plasma of CAP patients.MATERIAL AND METHODS:Blood plasma and erythrocytes obtained from: 42 patients with moderate severity pneumonia, 12 patients with grave severity pneumonia and 32 healthy volunteers. Content of advanced oxidation protein products, malondialdehyde and reactive carbonyl derivatives were estimated as indicators of the oxidative stress and oxidative damage of proteins.RESULTS:In patients with grave severity the level of oxidative proteins and MDA in erythrocytes exceeded both: control values and similar meanings in CAP patients with moderate severity. The further growth of MDA in this group patients’ blood plasma was observed, but the level of oxidative proteins decreased in comparison with those in CAP patients with moderate severity.CONCLUSION:To sum up, our derived data show, that injury of erythrocytes’ redox-status and blood plasma components plays an essential role in development and progression CAP.

Effect of freezing rate for cryopreservation of Persian sturgeon (Acipenser persicus) spermatozoa

This study examined the effect of freezing rate (−10 °C, −15 °C, −20 °C, −30 °C, and −40 °C/min) on motility parameters, rates of fertilization and hatching, ATP content, and indices of oxidative stress including thiobarbituric acid reactive substances and carbonyl derivatives of proteins in Persian sturgeon (Acipenser persicus) sperm. After sampling, sperm was diluted in an extender composed of 23.4-mM sucrose, 0.25-mM KCl, and 30-mM Tris-HCl, pH 8.0, containing 10% methanol and subsequently frozen in a programmable freezer. For postthaw sperm that were frozen at a rate of −40 °C/min, sperm motile duration (134 ± 27.01 seconds), sperm motile percent (60 ± 4.1%), fertilizability (72 ± 8.36% for fertilization rate and 65 ± 7.58% for hatching rate), and ATP content (4.8 ± 0.57 nmol/108 sperm) were significantly higher than for sperm frozen at any of the four slower rates (P < 0.05). Moreover, sperm cryopreserved using the fastest freezing rate had significantly lower levels of thiobarbituric acid reactive substances (0.5 ± 0.05 nmol/108 sperm) and carbonyl derivatives of proteins (41.3 ± 4.9 nmol/108 sperm) than sperm cryopreserved using all other freezing rates (P < 0.05). In addition, there is a significant difference (P < 0.05) between fresh sperm and the recovery of cryopreserved Persian sturgeon sperm using programmable freezing with −40 °C/min being the optimal freezing rate among those tested.

Urinary Markers for Early Detection of Diabetic Nephropathy in Type 1 Diabetes Mellitus

Background: diabetic nephropathy (DN) is a serious complication of diabetic mellitus associated with increased risk of morbidity and mortality. Diagnostic markers to detect DN at early stage are important as early intervention can slow loss of kidney functions and improve patient outcomes. N-acetyl-β-d-glucosaminidase (NAG) is a lysosomal enzyme, present in high concentrations in renal proximal tubular cells, Gamma-glutamyltransferase (GGT) is an enzyme which located along the proximal tubular brush border, Malondialdehyde (MDA) is a highly toxic product, formed in part by lipid oxidation derived free radicals, Reactive carbonyl derivatives (RCDS) is an oxidative stress marker in urine, as a measure of the oxidative modification of proteins and bete-2-microglobulin is filtered by the glomerulus, absorbed and catabolized by the proximal tubules. The aim of this study is to investigate the urinary outcome of these markers as early detectors of diabetic nephropathy in type 1 diabetic children. Subjects and methods: This case-control study included 67 children with type 1 diabetes mellitus (33 male; 34 female), age (11.03±1.05 years) and thirty one age (10.58±1.11 years) and sex (13 male; 18 female) matched healthy children (13 male; 18 female). Type 1 diabetic children were further subdivided into microalbuminuric and normoalbuminuric subgroups according to microalbuminuria concentration (30 mg/ g creatinine). Age, sex, diabetic duration and the current daily insulin dose, and family history of diabetes, weight, height, body mass index, systolic and diastolic blood pressure were recorded. Fasting plasma glucose, glycated hemoglobin, blood urea nitrogen, plasma creatinine, urinary creatinine, micoalbumin, N-acetyl-B-D glucosaminidase (NAG), Gama glutamyl transferase (GGT), Beta-2-microglobulin, Malondialdehyde (MDA) and Reactive carbonyl groups (RCDSS) were measured in all subjects. Results: a significant increase in tubular injury markers of diabetes (NAG, GGT, beta-2-microglobulin) and oxidative stress parameters (MDA, RCDSS) as compared to control subjects was found. Microalbuminuric subjects showed a significant elevatation in the urinary markers including NAG, GGT, beta-2-microglobulin, MDA, RCDSS as compared to normoalbuminuric subjects. The studied urinary tubular enzymes (NAG, GGT), oxidative stress markers (MDA, RCDSS) and Beta-2- microglobulin showed positive correlations with one another. Conclusion: The results of this study introduced the possibility of depending on tubular enzymes (NAG, GGT), oxidative stress markers (MDA, RCDSS) and β2 microglobulin as early, reliable, and sensitive predictors for diabetic nephropathy. The NAG activity index proved to be the most sensitive biomarker, then beta-2- microglobulin for early discovering the tubule cells damage.

Pharmacogenetics may Influence Tacrolimus Daily Dose, but not Urinary Tubular Damage Markers in the Long-Term Period after Renal Transplantation.

SummaryBackgroundThe primary goal of this study was to evaluate the influence of cytochrome P450 (CYP) 3A5 (6986A>G) and ABCB1 (3435C>T) polymorphisms on tacrolimus (TAC) dosage regimen and exposure. Second, we evaluated the influence of TAC dosage regimen and the tested polymorphisms on renal oxidative injury, as well as the urinary activities of tubular ectoenzymes in a long-term period after transplantation. Also, we aimed to determine the association between renal oxidative stress and tubular damage markers in the renal transplant patients.MethodsThe study included 72 patients who were on TAC based immunosuppression. Allele-specific PCR was used for polymorphism determination. We measured the urinary thiobarbituric acid reactive substances (TBARS) and reactive carbonyl derivates (RCD) in order to evaluate oxidative injury, as well as the urinary activities of ectoenzymes (N-acetyl-β-D-glucosaminidase, aminopeptidase N and dipeptidyl peptidase IV) to evaluate tubular damage.ResultsThe carriers of CYP 3A5*1 allele required statistically higher daily doses of TAC than CYP *3/*3 carriers, as well as the carriers of C allele of ABCB1 gene compared to those with TT genotype. Also, there were no differences in TBARS, RCD and the activities of ectoenzymes between the patients’ genotypes. Our results showed significant correlations between urinary TBARS and RCD and the ectoenzymes’ activities.ConclusionsOur findings suggest that CYP 3A5 and ABCB1 3435 polymorphism may affect TAC daily doses, but not the drug’s tubular toxicity. Furthermore, tubular damage may be associated with increased renal oxidative stress.

PP54 - Oxidative stress in blood plasma of patients with community-acquired pneumonia

Aim The main purpose of our investigation was to study the concentration of oxidized proteins and malondialdehyde (MDA) in blood plasma from patients with community-acquired pneumonia (CAP). Methods Patients were divided into 2 groups. 29 patients with community-acquired pneumonia moderate severity and respiratory insufficiency of grade 2 were included in the 1-st group. The control group consisted of 32 healthy persons. All patients and healthy subjects had received the full information on probable inconveniences at the blood sampling before giving their written informed consent. The protein reactive carbonyl derivatives, advanced oxidative proteins products (AOPP), and malondialdehyde (MDA) were detected in blood plasma. Comparisons of the results obtained between patients and healthy persons were performed using non-parametric Mann-Whitney U-test (for independent variables). Results Our results showed the significant increase in AOPP (by 2 times, p Conclusion The data showed synchronous increase of AOPP and MDA in blood plasma from CAP patients. AOPP is considered to be formed from oxidized albumin, fibrinogen. The probability of reaction of AOPP with MDA and cross-linking increases. It promotes oxidative stress over time and contributes to hypoxia progression in CAP patients.

PP55 - Oxidized proteins in neutrophils from patients with community-acquired pneumonia

Aim Oxidized proteins are considered as oxidative stress markers. The role of different oxidaized proteins in the progression of community-acquired pneumonia (CAP) might be of great importance. The aim of the present study was to evaluate advanced oxidation proteins products (AOPPs) and protein reactive carbonyl derivatives concentrations in neutrophils from CAP patients. Methods 29 patients with community-acquired pneumonia moderate severity and respiratory insufficiency of grade 2 were included in the 1-st group. Patients were observed in a hospital during our research. The control group consisted of 32 healthy persons. All patients and healthy subjects had received the full information on probable inconveniences and complications at the blood sampling before giving their consent to participate. The protein reactive carbonyl derivatives were detected following the protocol of Levine R.L. et al. The AOPPs was measured in neutrophils following the protocol of Witko-Sarsat et al. Results There was significant difference between the AOPPs concentrations in neutrophils from CAP patients and the control group. AOPPs concentration in plasma of patients was higher compared with healthy ones (by 2.3 times, p Conclusion Our results showed significant accumulation of AOPPs in neutrophils from CAP patients. We supposed that protein reactive carbonyl derivatives accumulation in neutrophils might be associated with further progression of pneumonia. In any case, accumulation of oxidized proteins might be connected with decrease of the proteolytic degradation of damaged proteins. In neutrophils excess accumulation of oxidized proteins could lead to misfolding and functional disorders of intracellular proteins, participating in cytoskeletal architecture, transport, redox homeostasis. It might be new insight systemic mechanism of CAP progression.