Proteoglycan/glycosaminoglycan and collagen content in the arterial wall of patients with end-stage renal disease: new indicators of vascular disease.

INTRODUCTION: The prevalence of cardiovascular (CV) comorbidity in patients with chronic kidney disease (CKD) is high, particularly in end­stage renal disease (ESRD). There is an ongoing search for novel biomarkers of CV disease in this population. OBJECTIVES: We aimed to investigate the associations of matrix proteoglycans (PGs) and glycosaminoglycans (GAGs), collagen, and arterial calcifications with selected serum and plasma markers of endothelial dysfunction, inflammation, oxidative stress, and bone turnover in patients with ESRD. PATIENTS AND METHODS: We enrolled 47 adult patients (32 men) with stage 5 CKD. The following parameters were investigated: fibrinogen, soluble thrombomodulin (sTM), plasminogen activator inhibitor 1 (PAI­1), stromal cell­derived factor 1α (SDF­1α), calcium (Ca), phosphate (Pi), intact parathormone, interleukin 6, high­sensitivity C­reactive protein (hs­CRP), ferric reducing ability of plasma, 2,2­diphenyl­1­picrylhydrazyl scavenging, ferric reducing ability of ascorbate in plasma, fetuin­A, fibroblast growth factor 23, osteopontin, osteoprotegerin, osteocalcin, transforming growth factor ß (TGF­ß), hepatocyte growth factor, secreted protein acidic and rich in cysteine, as well as matrix metalloproteinase 2. Radial artery specimens were stained with alizarin red for calcifications, alcian blue for PGs and GAGs, and sirius red for collagen. RESULTS: We observed positive correlations between PG and GAG, collagen, and calcification staining. The most intense (grade 3) alcian blue staining was significantly correlated with diabetes as well as higher levels of Ca × Pi product, hs­CRP, fibrinogen, SDF­1α, PAI­1, and sTM. However, PAI­1 was the only significant predictor of grade 3 alcian blue staining in a multiple logistic regression model adjusted for hemodialysis, Ca× Pi product, and hs­CRP levels. CONCLUSIONS: Coagulation disorders and endothelial dysfunction are the hallmarks of ESRD. The levels of SDF­1α, PAI­1, sTM, and fibrinogen may be novel predictors of early vascular wall alterations and may serve as CV risk markers.

Relationship between dietary antioxidant index (DAI) and antioxidants level in plasma of Kraków inhabitants.

BACKGROUND: Some literature data indicate that antioxidant-rich food may significantly increase antioxidants in serum and decrease the oxidative stress but results are ambiguous. OBJECTIVES: The aim of this study was to estimate the total antioxidant capacity of food intake among the inhabitants of Kraków, Poland on the basis of dietary antioxidant index (DAI) and evaluation the relation between DAI and the level of antioxidants in plasma. MATERIAL AND METHODS: Examination included 70 (37 women and 33 men) non-smoking inhabitants of Krakow aged 46.4 ± 13.7 years. DAI was investigated on the basis of Food Frequency Questionnaire including 145 food items. DAI was measured using the method by Benzi and expressed as FRAP (mMol/L). In plasma samples total antioxidant status (TAS) expressed as FRAP and malondialdehyde (MDA) concentration as a marker of lipids peroxidation were measured. RESULTS: The mean value of DAI of all examined persons was 46.74 ± 25.5 mMol/L (in female group: 54.13 ± 27.7 mMol/L; in male group: 37.83 ± 19.5 mMol/L; p < 0.05). The highest contribution in total DWA value had fruits (48.7%) opposite to vegetables (9.3%). Statistically significant positive correlations between DAI and FRAP in plasma was found in all: r = 0.42 and in female: r = 0.54 groups (not significant in men group: r = 0.20). Statistically significant negative correlation of DAI with MDA (malonylaldehyde) in female (-0.49) and male (-0.51) groups. CONCLUSIONS: The obtained results confirmed the hypothesis that the intake of antioxidants in daily diet (measured as DAI) might increase antioxidants defense (measured by TAC as FRAP) and decrease oxidative stress (measured by MDA concentration in plasma). The dietary modification towards higher consumption of antioxidants (especially in men) should be highlighted in prevention of diseases in which oxidative stress play considerable role.

Effect of Micronized Purified Flavonoid Fraction Therapy on Endothelin-1 and TNF-α Levels in Relation to Antioxidant Enzyme Balance in the Peripheral Blood of Women with Varicose Veins.

OBJECTIVE: The aetiology of varicose veins involves various factors and pathomechanisms including endothelial cell activation or dysfunction, venous hypertension, vein wall hypoxia, shear stress disturbances, inflammatory reaction activation or free radical production. To improve our understanding of the mechanisms of potential pharmacological interventions for chronic venous disease, we evaluated the influence of micronized purified flavonoid fraction (MPFF) on the relationship between antioxidant enzyme balance, endothelin-1 (ET-1) and tumour necrosis factor-α (TNF-α) levels. MATERIAL AND METHODS: Blood samples were obtained from 89 women with primary varicose veins; 34 were treated with MPFF and 55 did not receive any phlebotropic drug treatment. For the evaluation of the blood antioxidant enzyme balance, catalase (CAT) and superoxide dismutase (SOD) activity was assessed and the CAT/SOD ratio was calculated. RESULTS: Patients taking MPFF had significantly lower ET-1 levels than those not taking MPFF [median (25-75th quartile): 24.2 (22.30-27.87) vs 37.62 (24.9-44.58) pg.ml-1; p <0.05]. In those taking MPFF, a higher CAT/SOD ratio [39.8 (24.7-72.6) vs 28.8 (16.3-57.7); p<0.05] and a lower TNF-α concentration [6.82 (4.42-13.39) vs 12.94 (6.01-27.33) pg.ml-1; p<0.05] was also observed. In women not taking MPFF, ET-1 levels increased with the CAT/SOD ratio. In those taking MPFF, the ET-1 level was stable at approximately 25.0 pg.ml-1¬ up to a CAT/SOD ratio of 100. TNF-α level increased continuously with an increasing CAT/SOD ratio; however, the highest levels of TNF-α were observed in women not taking MPFF. CONCLUSION: We demonstrate the ability of MPFF to effectively lower the levels of ET-1 and TNF-α in patients with chronic venous disease. Further investigations are needed to define the therapeutic potential of MPFF including the potential effect on chronic subclinical inflammation, antioxidant imbalance and vascular dysfunction during the development of chronic venous disease.

Cardiovascular risk in chronic kidney disease patients: intima-media thickness predicts the incidence and severity of histologically assessed medial calcification in radial arteries.

BACKGROUND: The objective of the study was to determine the relationship between common carotid artery intima-media thickness (CCA-IMT) and histologically assessed calcification of radial artery in relation to clinical features and laboratory markers of bone and mineral metabolism, inflammation, and oxidative stress in patients with stage 5 chronic kidney disease (CKD). METHODS: The study comprised 59 patients (36 hemodialyzed, 23 predialysis). CCA-IMT was measured by ultrasonography; the biochemical parameters examined were assessed using routine laboratory methods, ELISA micro-plate immunoassays and spectrophotometry. Fragments of radial artery obtained during creation of hemodialysis access were cryosectioned and stained for calcifications using von Kossa method and alizarin red. RESULTS: Glucose, osteoprotegerin, pentraxin 3 and Framingham risk score significantly correlated with CCA-IMT. In multiple regression analysis, OPG positively predicted CCA-IMT. Radial artery calcifications were found in 34 patients who showed higher CCA-IMT (0.98 ± 0.13 vs 0.86 ± 0.14 mm; P = 0.006). Higher CCA-IMT values were also associated with more advanced calcifications. CCA-IMT and the presence of plaques in common carotid artery were positive predictors of radial artery calcifications, independent of dialysis status, Framingham risk score, CRP and Ca x Pi [OR for calcifications 2.19 (1.08-4.45) per 0.1 mm increase in CCA-IMT]. The presence of radial artery calcifications was a significant predictor of mortality, independent of dialysis status and Framingham risk score [HR 3.16 (1.03-9.64)]. CONCLUSIONS: In CKD patients, CCA-IMT examination can be used as a surrogate measure to assess the incidence and severity of arterial medial calcification which is associated with poor clinical outcome in these patients.

Vascular effects of advanced glycation end-products: content of immunohistochemically detected AGEs in radial artery samples as a predictor for arterial calcification and cardiovascular risk in asymptomatic patients with chronic kidney disease.

OBJECTIVES: Our aim was to determine whether vascular deposition of advanced glycation end-products (AGEs) is associated with arterial calcification and cardiovascular mortality in chronic kidney disease (CKD) patients and to assess the relationships between vascular content of AGEs and selected clinical and biochemical parameters. MATERIALS AND METHODS: The study comprised 54 CKD patients (33 hemodialyzed, 21 predialyzed). Examined parameters included BMI, incidence of diabetes, plasma fasting glucose, AGEs, soluble receptor for AGEs and 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging, serum C-reactive protein (hsCRP), plasminogen activator inhibitor-1 (PAI-1), and fetuin-A. Fragments of radial artery obtained during creation of hemodialysis access were stained for calcifications using alizarin red. AGEs deposits were identified immunohistochemically and their relative content was quantified. RESULTS: Vascular content of AGEs was positively correlated with BMI, hsCRP, fetuin-A, PAI-1, and DPPH scavenging in simple regression; only fetuin-A was an independent predictor in multiple regression. There was a significant positive trend in the intensity of AGEs immunostaining among patients with grades 1, 2, and 3 calcifications. AGEs immunostaining intensity predicted 3-year cardiovascular mortality irrespective of patient's age. CONCLUSIONS: The present study demonstrates an involvement of AGEs in the development of medial arterial calcification and the impact of arterial AGE deposition on cardiovascular mortality in CKD patients.

[Diet with high antioxidant capacity as important factor in primary and secondary prevention of asthma]. / Dieta o wysokim potencjale antyoksydacyjnym jako wazny czynnik w pierwotnej i wtórnej prewencji astmy oskrzelowej.

UNLABELLED: In primary and secondary prevention of diseases resulted from antioxidant failure and changes towards oxidation processes (autoimmunological/allergic diseases, especially asthma) intake of exogenic antioxidants from food is crucial. The aim of the study was to estimate diet antioxidant capacity (measured as DWA values) among healthy inhabitants of Krakow and assess which food products had the main contribution in DAI, in order to give the indications for people suffering from bronchial asthma. MATERIAL AND METHODS: The study group consisted of 478 persons: 350 of healthy inhabitants of Krakow and two specific subpopulations: 48 men with high level of physical activity and 80 students from Jagiellonian University and Technical Academy. Dietary antioxidant index (DWA) was investigated on the basis of food frequency questionnaire (FFQ). In the FFQ 145 food items were classified into groups such as fruit and dried fruits, vegetables and vegetable oils, sweets, grains and cereal-based products, mixed dishes, fresh herbs, spices and beverages. The total antioxidant capacity of food intake was measured using the method developed by Benzi. This method exploits the ability of the sample to reduce ferricions to ferrousions (FRAP). RESULTS: The total antioxidant capacity (measured as DWA) of diet of 225 examined women was higher than calculated from 253 men and statistically significant (82.7 Mmol/l vs 49.1 Mmol/l). Moreover FRAP values of particular groups of food products were higher in case of women than men. The highest contribution in total DWA value had fruits and subsequently: cereals, juices and beverages. Unfortunately, examined people consumed to low amount of vegetables, fresh herbs and spices, which have well-documented antioxidant property. CONCLUSIONS: DWA values of healthy individuals presented in this study may be treated as a low reference value of antioxidant capacity for diets of asthmatic people. Enrichment in natural antioxidants diets of asthmatic individuals (mainly men), should become an important element of primary and secondary prophylaxis in bronchial asthma.

Detection of potential chronic kidney disease markers in breath using gas chromatography with mass-spectral detection coupled with thermal desorption method.

The analytical potential of chromatographic breath analysis towards detection of compounds suggested as markers of chronic kidney disease (CKD) was tested. Until now, trimethylamine (TMA) considered as a potential marker of renal disorder was detected mainly in plasma. Detection of TMA in breath was rarely undertaken due to analytical difficulties associated with amines' properties. The results of our investigations confirmed that an application of thermal desorption (TD) and gas chromatography with mass-spectral detection (GC/MS) allows direct detection of TMA in breath. The preliminary studies allowed to determine the breath composition in case of patients suffering from CKD and to compare the obtained results to a control group. Breath samples were collected from 14 patients and 9 healthy volunteers. TMA was detected in all patients suffering from CKD in the range 1.76-38.02ppb, but not in the control group. Acetone and isoprene were present in the exhaled air of all examined persons. The concentration of acetone was in the range of 26.52-329.46ppb in the patient group and 73.11-437.14ppb in the control group. Isoprene was detected in the range 57.17-329.8ppb among CKD patients and 27.99-143.77ppb in healthy volunteers. Additionally aliphatic hydrocarbons and sulfur compounds were determined in breath as compounds which could be essential in case of diseases coexisting with CKD. Apart from TMA and pentane no statistically significant differences were found using our analytical technique. TMA was detected in the breath of all patients with CKD and in none of breath samples in control group. TMA seems to be a promising marker of CKD.

[New opportunities of renal diseases diagnostics using gas chromatography]. / Nowe mozliwosci diagnostyki chorób nerek przy wykorzystaniu chromatografii gazowej.

During the last decade the amount of patients suffering from chronic kidney disease (CKD) has increased. The physicians' efforts are focused on early CKD diagnosis and reduction of the end-stage renal diseases incidence. The breath test seems to be a promising diagnostic device offering early noninvasive diseases detection. The aim of presented study was the determination of breath composition in case of persons suffering from CKD. Breath samples were collected from 14 patients and 7 healthy volunteers. Exhaled air samples were analyzed by gas chromatography equipped with mass spectrometer (GC-MS). Samples were enriched using solid phase microextraction (SPME). Trimethylamine (TMA), mentioned in literature as potential marker of chronic kidney diseases, was detected in case of 11 patients. Among breath components were detected: sulfur compounds: dimethyl sulfide (was detected in exhaled air of patients and control group), carbon disulfide (was detected in case of 4 healthy and in case of all patients) and also potential markers of oxidative stress: propane, butane, pentane, 2-methylpentane, hexane. Acetone and isoprene occurred in exhaled air of all studied persons. The considerable increase of acetone concentration in comparison to control group was observed in case of patients with diagnosed diabetes. The application of gas chromatography with mass spectrometer and appropriate enrichment of samples allows to define the breath profile characteristic for chosen the unit of disease. Typical compounds--biomarkers can be useful for early diagnostics.

[Obesity and its' influence on reactive oxygen species (ROS) in the blood of patients with varicose veins of the lower limbs]. / Wplyw otylosci na poziom reaktywnych form tlenu (RTF) we krwi chorych z zylakami konczyn dolnych.

INTRODUCTION: Oxidative stress is considered as a probable mechanism in the pathogenesis and development of many diseases, including chronic venous disease (CVD). Local increased production of reactive oxygen species is taken into account as a mediator in vessel wall changes leading to damage of the endothelium and may be the mechanism leading to its' decreased blood flow and venous stasis. Obesity is a known clinical factor influencing venous blood flow from the lower limbs. AIM: Research aim was to evaluate the local production of ROS on the basis of malonylodialdehyde (MDA) production--a product of lipid peroxidation in blood samples taken from varicose veins of the lower limbs; as well as from blood taken from the forearms of patients that undertook surgery for lower limb varicose veins. The correlation between MDA level and body mass index (BMI) was also examined. MATERIALS AND METHODS: The examined group consisted of 31 patients with diagnosed CVD of the 2nd and 3rd degree according to CEAP qualified for surgical procedure. The analyzed group consisted of 23 females and 8 males between the ages of 26-68 years (mean age 49.9 +/- 12.5). The control group included 31 patients (20 females and 11 males) between the ages of 51-72 years (mean age 60 +/- 12); volunteers were without signs and symptoms of CVD whom had MDA levels measured from forearm veins during routine exam. Lipid peroxidation products were identified by measuring reactive species reaction with thiobarbital, measured by the spectro-fluorometrical method given by Austa. MDA concentration were expressed as nmol of MDA per ml of plasma and measured using the fluorometric method at fluorescence emission at 553 nm and ebullition at 532 nm. RESULTS: Significant increase in the MDA level in blood with various veins was noted in comparison to peripheral blood in patients with signs and symptoms of CVD (p < or = 0.0001). Overall MDA concentrations was higher in venous blood plasma in patients with CVD than in healthy volunteers (p < or = 0.0005). BMI substantially correlated with increased MDA concentration in blood plasma. MDA concentration was higher in obese patients than in patients with normal BMI. These results were substantially significant (p < or = 0.001). CONCLUSIONS: CVD is associated with increased oxidative stress measured by the level of MDA in blood plasma. Measurement of MDA may be a useful marker in evaluation of vessel changes in patients with CVD. 3. Obesity increases risk of lipid peroxidation and influences increment of oxidative stress in the group of patients with CVD.

Prolonged quercetin administration diminishes the etoposide-induced DNA damage in bone marrow cells of rats.

The DNA damage in bone marrow cells induced by etoposide (E) injected intraperitoneally to rats (100 mg/kg b.w.) decreased to the control level when quercetin (Q) was administered subcutaneously for 10 consecutive days (40 mg/kg b.w.per day) before E was injected. The antioxidant power (FRAP assay) increased significantly after Q or E compared with control rats but did not change when Q preceded the E injection. The superoxide dismutase activity significantly increased in Q+E-treated rats compared with quercetin given alone. The study provides evidence that Q protects bone marrow cells against long-lived E-induced DNA damage and alters the redox balance in lung tissue.

The usefulness of artificial neural networks in the evaluation of pulmonary efficiency and antioxidant capacity of welders.

OBJECTIVE: The aim of the study was to determine if artificial neural networks (ANNs) may be useful to analyse a complex and large set of data derived from smoking welders for the purpose of finding relationships between parameters describing respiratory system efficiency and antioxidant defence. METHODS: A group of 94 welders employed in a big metallurgic enterprise in Krakow, Poland (men only, aged 29-57 years, all active smokers) occupationally exposed to O(3) and NO(x), were the subjects of this study. They underwent biochemical measurements including total antioxidant status (TAS) and the anti-oxidative defence enzymes superoxide dismutase (SOD) and catalase (CT); biominerals: Fe, Cu, Zn, Mg in blood serum and in hair; the concentrations of albumin, bilirubin, uric acid in blood. The determination of respiratory efficiency was based on a "flow-volume" curve and spirometry. The dependant variables for ANNs were: TAS, SOD, CT. Thirty-one subjects with normal values of all spirometric parameters were selected for the final analysis. RESULTS: Statistically valid relationship between TAS and albumin, Zn and Cu in blood and the two pulmonary parameters forced expiratory volume after 1s (FEV(1)) and middle expiratory flow of 25-75% of vital capacity (MEF(25/75)) were found. Zn concentration almost linearly influenced TAS. For Cu a sigmoid curve was obtained. For albumin concentration as well as for FEV(1) a two-stage curve was observed. CONCLUSIONS: ANNs are useful for the reduction of dimensionality and are suited to analyse a complex and relatively large set of parameters when it is unknown which of these are related. ANNs were useful for finding the relationship between the antioxidant defence and the respiratory system capacity in welders who smoke.

Antioxidant enzymes and pulmonary function in steel mill welders.

It is known that high levels of nitric oxide and ozone lead to disturbances of the balance between oxidants and antioxidants. The purpose of this study was to investigate ventilatory parameters in relation to the antioxidant status measured as total antioxidant status (TAS), superoxide dismutase (SOD) and catalase (CT). The study group consisted of 94 welders, aged 41.2 +/- 10.0 years, employed in the Steel Mill in Kraków, Poland, and exposed to nitric oxides and ozone in concentrations exceeding the threshold limit values. The control group consisted of 115 unexposed healthy workers aged 40.8 +/- 10.2 years. All the subjects under study were smokers. Determination of ventilatory efficiency was based on a "flow-volume" curve and spirometry. TAS was measured using reagents from the Randox Laboratories Ltd, SOD according to Fridovich and CT with Aebi's method. It was found that in the group of welders, the concentrations of TAS, CT and SOD were lower compared to controls (TAS-1.15/1.33 mmol/ml; CT-18.1/28.4 m/gHb, SOD-767.6/855.6 U/gHb). The incidence of extreme obstructive pulmonary disease and small airway disease in the welder group was more frequent than in controls. Changes in the concentration (or activity) of antioxidant parameters cannot be used as early markers of ventilatory dysfunction, although the values in the lowest class of TAS, SOD and CT showed a significantly larger number of welders than controls.