Neutrophil Oxidized-Modified Proteins and Neutrophil Extracellular Traps in Patients with Community-Acquired Pneumonia.

MATERIALS AND METHODS: 51 patients with CAP were divided into 2 groups depending on the severity of the pathological process. The first group (I) consisted of 32 patients with moderate severity of pneumonia. The second group (II) consisted of 19 patients with severe pneumonia. The third group (III), the comparison group, consisted of 14 CAP patients with chronic obstructive pulmonary disease (COPD). The control group consisted of 19 volunteers. RESULTS: Statistically significant increase in the level of carbonyl derivatives (CD) in patients of all study groups relative to the control group was revealed. In the group of patients with moderate severity and severe pneumonia, also in CAP patients with COPD, the level of CD exceeded the control group. There was no statistically significant difference in the level of advanced oxidation protein products (AOPP) and myeloperoxidase (MPO) in blood neutrophils between the studied groups. CONCLUSION: Results indicate an oxidative imbalance in neutrophils and contribute to the worsening of the course of the disease.

Dynamic changes in purine catabolism in patients with acute coronary syndrome that underwent percutaneous coronary intervention.

BACKGROUND: Cardiovascular diseases are global problems. They are causes of death in about 43% of people worldwide and may become the most widespread reason of death by 2020. The prognosis is directly dependent to immediate diagnosis and on time treatment. Introduction of new biochemical markers as the early diagnosis of complications after coronary revascularization is very important in this period. Herein, we assayed the changes of purine catabolites in patients with acute coronary syndrome (ACS) before and after percutaneous coronary intervention (PCI) in comparison with control group. METHODS: Thirty five ACS patients (20 males and 15 females) were included (57±17 years old) in the study. The determination of intermediates of purine catabolism as guanine, hypoxanthine (GCS), adenine, xanthine (Kc) and uric acid (MK) were assayed before and 3 days after PCI. Conditionally, 35 healthy-matched persons were included in the control group. Purine catabolites were determined in plasma through the method of Oreshnikov E.V (2008). RESULTS: In ACS patients, prior to PCI, there was a tendency to increase the concentration of guanine (P=0.001), hypoxanthine (P=0.002) adenine (P=0.0003), xanthine (P=0.000003) and uric acid (P=-0.000001) relative to the upper limits of normal ranges. And on the third day after PCI, there was the second tendency to increase the levels of guanine (P=0.000001), hypoxanthine (P=0.000001) adenine (P=0.0000001), xanthine (P=0.000001) and uric acid (P=0.0000001) relative to upper limits of normal ranges. CONCLUSION: Increment of plasma purine catabolites can be a marker of inflammation and instability of coronary artery plaques and may be used as a restenosis marker in patients with history of PCI.

The Oxidized proteins and activity of the Cl-/HCО3- exchanger in erythrocytes of patients with acute alcohol intoxication.

BACKGROUND: 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. METHODS: In erythrocytes the activity of Cl-/HCО3- exchanger, the reactive protein carbonyl derivatives and membrane-bound hemoglobin concentration were measured. RESULTS: 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. CONCLUSIONS: 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.

Influence of Combined Therapy on Generation of Neutrophil Extracellular Traps in Patients with Cervical Cancer.

BACKGROUND: According to several authors, neutrophil extracellular traps (NETs) play an important role in the mechanisms of cancer development and metastatic processes, which allows them to be considered as a potential new target for the treatment of cancer. AIM: To investigate the presence of extracellular neutrophil traps in the blood of patients with cervical cancer on the background of the combined treatment. MATERIALS AND METHODS: The study was conducted in 28 patients with cervical cancer. Group 1 received only radiation therapy; Groups 2-radiation therapy with ftorafur; Group 3-radiation therapy with cisplatin. To determine the number of spontaneous extracellular neutrophilic traps in the blood of the examined individuals, we used a technique of I.I. Dolgushin and Yu.S. Andreeva. RESULTS: Peripheral blood neutrophils in 53.57% (33.87; 72.49) of cervical cancer patients showed the ability to generate NETs before treatment. The ability to form NETs was observed in neutrophils isolated from 66.67% (9.43; 99.16) patients of the Group 1. After radiation therapy with ftorafur, the ability of blood neutrophils to form NETs was observed in 50% (1.26; 98.74) of cervical cancer patients. After radiotherapy with cisplatin, 37.50% (15.20; 64.57) of patients were found to have NETs formation. CONCLUSION: The ability to form NETs varied greatly after radiotherapy. The addition of chemotherapy drugs to radiation therapy did not increase the percentage of NETs in the blood of patients with cervical cancer but stimulated the appearance of basophil extracellular traps.

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.

The modified proteins in erythrocytes at hemodialysis patients.

The main goal of our research was to study the modified proteins and activity of integral band 3 protein in erythrocytes of patients with end-stage renal failure during hemodialysis. Depending on the starting nosological variant 2 groups were formed. The first group included 27 patients with end-stage renal failurein the outcome of chronic pyelonephritis, the second - 20 patients with end-stage renal failure, developed on a background of chronic glomerulonephritis. The control group consisted of 15 healthy subjects. The patients of 1-th and 2-nd groups were treated with hemodialysis. The concentration of reactive protein carbonyl derivates, membrane-binding hemoglobin were detected in erythrocytes. Activity of band 3 protein was detected following the protocol of I. Mindukshev et al. (2010). Comparison the results obtained was performed using non-parametric Mann-Whitney U-test (for independent variables). Before hemodialysis the significant increasing of concentration of reactive protein carbonyl derivates and membrane-binding hemoglobin in erythrocytes of both group patients in comparison with healthy ones was observed. During hemodialysis the further augmentation of modified proteins in erythrocytes took place. Different changes in the activity of the protein band 3 in erythrocytes of patients during hemodialysis were found. The similar type of protein band 3 activity changing was determined in erythrocytes of 1-th group patients. There was positive pair correlation between concentration of the reactive protein carbonyl derivates and protein band 3 activity (r= 0,31, p<0,05). Two different types of protein band 3 activity changing were determined in erythrocytes of 2-th group patients. Thus, the differences in modified proteins concentration and protein band 3 activity that occurred in the erythrocytes of patients during hemodialysis were established.

Characteristic of oxidative metabolism in patients with chronic kidney disease depending on etiological nosology.

Development of chronic kidney disease (CKD) is accompanied by change in the oxidative metabolism. The aim of the study was to assess the parameters of oxidative stress in erythrocytes and plasma of patients with CKD 1 and 2 stages depending on the etiological nosology. Two groups were formed. 1(st) - patients with chronic glomerulonephritis (CG), CKD 1, 2. 2(nd) - patients with chronic pyelonephritis (CP), CKD 1, 2. The concentration of carbonyl derivatives was determined by the method of Levine R.L. Malondialdehyde (MDA) concentration - by the method of Goncharenko M.S. and Lapytova A.M. Median and quartiles of parameters were calculated. Statistically significant difference of data in the comparison groups were assessed using the Mann-Whitney test. In group of patients with CG there is the tendency to reduce the concentration of carbonyl derivatives in erythrocytes Md=12.861nmol/ml (12.468, 15.177) as compared with patients with CP Md=14.346nmol/ml (14.130, 14.537). Significant differences in this indicator in these groups were not detected, p=0.715. The concentration of MDA in erythrocytes in these groups varied in the opposite way. In patients with CG concentration of MDA in erythrocytes Md=5.513 µmol/ml (4.487, 6.282) was higher than this indicator in patients with CP Md=4.744 µmol/ml (3.974, 5.256), these changes were not significant, p=0.256. The concentrations of carbonyl derivatives and MDA in plasma in the compared groups were almost identical. In patients with CG concentration of carbonyl derivatives in plasma was Md=0.390nmol/ml (0.381, 0.528), the content of MDA Md=0.846 µmol/ml (0.714, 1.005). In patients with CP concentration of carbonyl derivatives in plasma was Md=0.424nmol/ml (0.312, 0.485) (when comparing 2 groups p=0.668), the content of MDA Md=0.820 µmol/ml (0.740, 0.979) (p=0.886). Thus, in patients with CKD 1, 2 depending on the etiological nosology features of oxidative metabolism were not found.

Myeloperoxidase activity and the oxidized proteins in blood neutrophils of patients with pneumonia.

The main purpose of our investigation was to study myeloperoxidase activity and concentration of oxidized proteins in blood neutrophils of patients with ambulant pneumonia and secondary pneumonia which has arisen on a background of chronic obstructive pulmonary disease (COPD). Patients were divided into 2 groups. 17 patients with ambulant pneumonia moderate severity and respiratory insufficiency of grade 2 were included in the 1-st group. 20 COPD patients with secondary pneumonia moderate severity and with respiratory insufficiency of grade 2 were included in the 2-nd group. The control group consisted of 15 healthy subjects. The reactive protein carbonyl derivates, advanced oxidation protein products (AOPP) and myeloperoxidase activity were detected in neutrophils. In neutrophils of 1-st group patients the augmentation of reactive protein carbonyl derivates was observed in comparison with healthy ones. In neutrophils of 2-nd group patients the slight decrease of reactive protein carbonyl derivates was observed in comparison with healthy ones (by 17%). In neutrophils of 2-nd group patients the significant increasing AOPP in comparison with healthy ones (p <0.01) and 1 group patients (p <0.05) was fixed. Myeloperoxidase activity was higher in neutrophils of 1-th group patients in comparison with healthy ones. In neutrophils of 2-nd group patients myeloperoxidase activity was higher in comparison with the same of 1 group patients (by 67%, p <0.05). Our results showed the different direction of oxidized proteins formation neutrophils of patients with primary and secondary pneumonia. Besides that the varied degree of myeloperoxidase activity was fixed. Our results require more detailed understanding because they can reflect peculiar mechanisms of pneumonia development and determine the characteristics of their progression.