Maria Iskra

Analysis of serum copper and zinc concentrations in cancer patients

Several studies have shown that plasma copper concentrations are increased in various carcinomas. Zinc acts as a cellular growth protector, including growth of neoplastic cells, and its deficiency was demonstrated to be involved in several stages of malignant transformation. However, the usefulness of the serum zinc and copper determinations in cancer prevention, detection, monitoring treatment, and prognosis requires further investigations. The aim of the present study was to compare the serum copper and zinc levels in patients with cancer of the lung (PC), breast (BC), gastrointestinal tract (GIC), and gynecological (GYNC) malignancy with progress of the disease. The results of the study have shown a significant increase in the mean total serum Cu levels and the serum Cu/Zn ratio in all patient groups with cancer compared to a control group. Increased mean serum concentrations and Cu/Zn ratios were found in the whole group (ALLC), and for the GIC and GYNC groups with local as well as metastasized (Meta) disease in comparison with the control group. The mean serum concentrations of Zn were decreased only in metastasized ALLC and GYNC groups.

Copper and zinc concentrations and the activities of ceruloplasmin and superoxide dismutase in atherosclerosis obliterans.

The relationships among concentrations of copper and zinc, the oxidase activity of ceruloplasmin (Cp) in serum, and Cu,Zn-SOD (superoxide dismutase) activity in erythrocytes were investigated in men with atherosclerosis obliterans (AO) and a control group. The oxidase activity of Cp was measured with o-dianisidine dihydrochloride as a substrate, and Cu,Zn-SOD activity in erythrocytes by using the RANSOD kit. The lipid profile and uric acid concentration were determined in AO and control groups.The results showed higher copper and zinc concentrations in serum in the AO group (20.0±3.5 and 18.0±3.2 µmol/L, respectively) in comparison with the control group (15.6±2.3 and 14.7±1.9 µmol/L). The Cp activity in serum was higher in the AO group (174.2±61.8 U/L) than in the control group (93.7±33.9 U/L), and a significant difference was found in the activity of Cu,Zn-SOD in erythrocytes (2389±1396 and 1245±365 U/g Hb, respectively) between both groups.The activity of Cu,Zn-SOD was positively correlated with copper in the control group (r=0.73), but not in AO, and negatively with uric acid concentration (r=−0.63) in the AO group. The oxidase activity of Cp was correlated with copper, but not zinc, in AO and control groups (r≥0.65). Negative correlation coefficients were calculated for uric acid and copper and zinc concentrations in the AO group (−r≥0.61).Increased copper concentrations and oxidase activity of Cp in serum in AO and the activity of Cu,Zn-SOD in erythrocytes could result from atherosclerotic disease, accompanied by chronic ischemia of a lower limb. These results suggest also that relationship between copper concentration and Cu,Zn-SOD activity in erythrocytes found in the serum of healthy subjects may be disturbed in pathologic conditions.

Arylesterase and paraoxonase activity of paraoxonase (PON1) affected by ischemia in the plasma of patients with arterial occlusion of the lower limbs.

OBJECTIVES The aim of the study was to evaluate the effect of different degrees of chronic ischemia of the lower limbs on PON1 activity in plasma, in relation to different substrates. DESIGN AND METHODS The studied group consisted of patients with chronic arterial occlusion of the lower limbs due to atherosclerosis. The paraoxonase and arylesterase activities of PON1 were measured according to the Karen Gan method. RESULTS PON1 arylesterase activity was affected by ischemia of the lower limbs depending on its degree. In the group with critical ischemia decreased PON1 activity was observed in comparison with that in the moderate ischemia group and the control group (50.05+/-21.40 U/mL and 82.59+/-29.27 U/mL, 85.30+/-35.05 U/mL, respectively). CONCLUSIONS The results revealed a reverse relationship between PON1 activity and the progress of atherosclerosis to the ischemic level. The present study demonstrates, for the first time, that the arylesterase activity of PON1 is affected by critical ischemia of the lower limbs.

Serum iron concentration and plasma oxidant-antioxidant balance in patients with chronic venous insufficency.

Summary Background The aim of this study was to evaluate serum iron concentration and influence of this element on biomarkers of oxidative stress in patients affected by chronic venous insufficiency (CVI). Material/Methods Serum iron (SI) concentration and plasma parameters of oxidant–antioxidant balance (i.e., malonyldialdehyde [MDA], uric acid [UA] concentration, and total antioxidant capacity [TAC]) were compared between 35 patients divided into appropriate groups and 23 healthy individuals. Results The subgroups analysis showed that SI concentration was significantly higher only in patients with shorter duration of CVI in comparison with the control group (P=0.013). Significant, negative correlation was found between SI concentration and duration of the disease (r=−0.422, P=0.014), age of the patients (r=−0.542, P=0.001) and BMI (r=−0.408, P=0.018). Mean value of MDA concentration and TAC capacity were higher in patients with CVI in comparison with healthy individuals (P<0.05). UA concentration was decreased, especially in CVI patients with mild clinical stage of disease and shorter CVI duration (P=0.047; P=0.034). There was no significant correlation found between SI concentration and the parameters of oxidant–antioxidant balance. Conclusions High concentration of MDA and low UA level in blood of CVI patients suggests that oxidative stress plays an important role in the pathogenesis of the disease. The increase in SI concentration observed in the early stage of CVI can enhance free radicals formation; however, direct evidence has not been provided by the present study.

PON1 status evaluation in patients with chronic arterial occlusion of lower limbs due to atherosclerosis obliterans.

Introduction Human paraoxonase (PON1) is a calcium-dependent enzyme physically associated with HDL, and it is believed to contribute to the atheroprotective effect of HDL. The aim of the study was to evaluate PON1 status in patients with atherosclerosis obliterans as an effect of ischemia regarding its activity and phenotype distribution. Material and methods The study group consisted of patients with chronic arterial occlusion of the lower limbs due to atherosclerosis obliterans (AO). The patients were divided into two groups according to the degree of ischemia: moderate (MI), and critical (CI). The ratio of the hydrolysis of salt-stimulated PON1 activity to the hydrolysis of phenylacetate was used to assign individuals to one of three possible phenotypes (low activity – A, medium activity – AB, high activity – B). It was observed that PON1 arylesterase activity was affected by ischemia of the lower limbs depending on its degree. Results The odds ratio and the relative risk analysis showed that the patients with moderate ischemia are much more often characterized by phenotype A than by phenotype B. The low activity phenotype A occurs over twice as often in patients with chronic ischemia of the lower limbs as in individuals from the control group (OR = 2.125; 1.96 to 3.776, p = 0.0143). Conclusions This study presents the low activity phenotype A in relation to the risk of ischemia of the lower limbs due to atherosclerosis and shows the potentially important role of PON1 in conclusion of the process leading to intensification of ischemia degree.

Measurement of Free Radicals Using Electron Paramagnetic Resonance Spectroscopy During Open Aorto-Iliac Arterial Reconstruction

Background Aortic cross-clamping during abdominal aortic aneurysm (AAA) open repair leads to development of ischemia-reperfusion injury. Electron paramagnetic resonance spectroscopy (EPR) spin-trapping is a valuable method of direct measurement of free radicals. The objective of the study was to evaluate the results of EPR as a direct method of free radical measurement and degree of inflammatory response in open operative treatment of patients with AAA and aorto-iliac occlusive disease (AIOD). Material/Methods The study was performed on a group of 32 patients with AAA and 25 patients with AIOD scheduled for open repair. Peripheral venous blood for EPR spectroscopy and for SOD, GPx, ox-LDL, Il-6, TNF-alfa, CRP, and HO-1 were harvested. Selected parameters were established accordingly to specified EPR and immunohistochemical methods and analyzed between groups by Mann-Whitney U test and Wilcoxon matched-pairs signed-ranks test with Bonferroni correction. Results Free radicals level was correlated with the time of the aortic cross-clamping after the reperfusion of he first and second leg in AAA (r=0.7; r=0.47). ox-LDL in AAA decreased 5 min after reperfusion of the first leg (32.99 U/L, range: 14.09–77.12) and 5 min after reperfusion of the second leg (26.75 U/L, range: 11.56–82.12) and 24 h after the operation (25.85 U/L, range: 14.29–49.70). HO-1concentration increased to above the level before intervention 24 h after surgery. The activities of GPx and SOD decreased 5 min after the first-leg reperfusion in AAA. Twenty-four hours after surgery, inflammatory markers increased in AAA to CRP was 14.76 ml/l (0.23–38.55), IL-6 was 141.22 pg/ml (84.3–591.03), TNF-alfa was 6.82 pg/ml (1.76–80.01) and AIOD: CRP was 18.44 mg/l (2.56–33.14), IL-6: 184.1 pg/ml (128.46–448.03), TNF-alfa was 7.74 pg/ml (1.74–74.74). Conclusions EPR spin-trapping demonstrates temporarily elevated level of free radicals in early phase of reperfusion, leading to decrease antioxidants in AAA. Elevated free radical levels decreased 24 h after surgery due to various endogenous antioxidants and therapies.

Advanced Oxidation Protein Products and Carbonylated Proteins as Biomarkers of Oxidative Stress in Selected Atherosclerosis-Mediated Diseases

Objectives The main question of this study was to evaluate the intensity of oxidative protein modification shown as advanced oxidation protein products (AOPP) and carbonylated proteins, expressed as protein carbonyl content (C=O) in abdominal aortic aneurysms (AAA), aortoiliac occlusive disease (AIOD), and chronic kidney disease (CKD). Design and Methods The study was carried out in a group of 35 AAA patients and 13 AIOD patients. However, CKD patients were divided into two groups: predialysis (PRE) included 50 patients or hemodialysis (HD) consisted of 34 patients. AOPP and C=O were measured using colorimetric assay kit, while C-reactive protein concentration was measured by high-sensitivity assay (hsCRP). Results The concentration of AOPP in both AAA and AIOD groups was higher than in PRE and HD groups according to descending order: AAA~AIOD > HD > PRE. The content of C=O was higher in the PRE group in comparison to AIOD and AAA according to the descending order: PRE~HD > AAA~AIOD. Conclusions AAA, AIOD, and CKD-related atherosclerosis (PRE and HD) contribute to the changes in the formation of AOPP and C=O. They may promote modification of proteins in a different way, probably due to the various factors that influence oxidative stress here.

The Association of Serum Thrombomodulin with Endothelial Injuring Factors in Abdominal Aortic Aneurysm

Background. The aim of the present study was to evaluate the concentration of serum thrombomodulin (sTM) in the AAA patients and to examine its correlation with various factors which may potentially participate in the endothelial injury. Materials and Methods. Forty-one patients with AAA were involved and divided into subgroups based on different criteria. Concentration of sTM was measured using enzyme-linked-immunosorbent assay (ELISA). The results were compared with those obtained in 30 healthy age- and sex-matched volunteers. Results. The higher concentration of sTM was observed in AAA patients compared with those in controls volunteers [2.37 (1.97–2.82) ng/mL versus 3.93 (2.43–9.20) ng/mL, P < 0.001]. An elevated sTM associated significantly with increased triglycerides (TAG) [P = 0.022], cholesterol [P = 0.029], hsCRP [P = 0.031], and advanced glycation end products (AGEs) [P = 0.033]. Conclusions. The elevation of serum sTM level suggests that endothelial damage occurs in AAA pathogenesis. The correlations observed indicate that lipids abnormalities, inflammation, and oxidative stress may be involved in this destructive process.

Altered elastase—alpha1-antitrypsin balance in the blood of patients with chronic venous disease

Objectives Although leukocyte elastase is suspected to be involved in the damage of vein wall during chronic venous disease, the equilibrium between this protease and its inhibitor, alpha1-antitrypsin, has not yet been evaluated. The aim of the present study was to determine the relationship between leukocyte elastase and alpha1-antitrypsin, in the blood of patients with chronic venous disease. Patients and methods The concentration and the activity of leukocyte elastase along with the activity of alpha1-antitrypsin were evaluated in the blood of 55 chronic venous disease patients. The results were compared with those obtained in 33 healthy age and sex-matched volunteers. Results A significant decrease in the leukocyte elastase activity that correlated with an increased alpha1-antitrypsin activity was observed in the serum of patients with mild clinical symptoms of chronic venous disease. Conclusions The results of the study did not confirm a hypothesis about an important role of proteolytic activity of leukocyte elastase in the vein wall injury mechanism. They show that the leukocyte elastase–alpha1-antitrypsin balance is rather shifted toward antiprotease activity, especially in an early stage of chronic venous disease.

Plasma concentration of selected biochemical markers of endothelial dysfunction in women with various severity of chronic venous insufficiency (CVI)—A pilot study

Background Although the endothelial dysfunction is considered to be implicated in the pathogenesis of chronic venous insufficiency (CVI) the endothelial status in patients with venous disorders is still not fully evaluated. Therefore the aim of the study was to measure the concentration of selected markers of endothelial dysfunction: von Willebrand factor (vWf), soluble P-selectin (sP-selectin), soluble thrombomodulin (sTM) and soluble VE-cadherin (sVE-cadherin) in CVI women who constitute the most numerous group of patients suffering from venous disease. Materials and methods Forty four women with CVI were involved in the study and divided into subgroups based on CEAP classification. Concentration of vWf, sP-selectin, sTM and sVE-cadherin were measured and compared with those obtained in 25 healthy age and sex-matched women. Results It was found that the concentration of sTM increased and sVEcadherin decreased along with disease severity in CVI women. A significant rise of sTM was observed especially in CVI women, with the highest inflammation status reflected by hsCRP or elastase concentration, and in CVI women with a high oxidative stress manifested by an increased plasma MDA. A significant fall of circulating sVE-cadherin was reported in CVI women with moderate to highest intensity of inflammation and oxidative stress. There was no change in vWF and sP-selectin concentration at any stage of CVI severity. Conclusions The results of the present study demonstrate the presence of endothelial dysfunction in women suffering from CVI which seems to progress with the disease severity and may be associated with inflammation and enhanced oxidative stress.