The influence of fatty acids on theophylline binding to human serum albumin. Comparative fluorescence study.

Theophylline, popular diuretic, is used to treat asthma and bronchospasm. In blood it forms complexes with albumin, which is also the main transporter of fatty acids. The aim of the present study was to describe the influence of fatty acids (FA) on binding of theophylline (Th) to human serum albumin (HSA) in the high affinity binding sites. Binding parameters have been obtained on the basis of the fluorescence analysis. The data obtained for the complex of Th and natural human serum albumin (nHSA) obtained from blood of obese patients qualified for surgical removal of stomach was compared with our previous studies on the influence of FA on the complex of Th and commercially available defatted human serum albumin (dHSA).

Advanced protein glycation in psoriasis.

BACKGROUND: The aetiopathogenesis of psoriasis is very complex and has not been well known. Recently, it has been proposed that mechanisms dependent on free radicals may be involved in the development of this dermatosis. Also, psoriasis coincides with lipid disturbances, diabetes and diseases of cardiovascular system. However, the common mechanism connecting these diseases is still unknown. OBJECTIVES: The aim of this study was to measure concentration of peptides containing glycated residues (AGE-peptides) and IgG, IgA and IgM antibodies against carboxymethyllysine (anti-CML) and carboxyethyllysine (anti-CEL) in the sera of patients at different phases of psoriasis activity in comparison with the sera of healthy individuals. METHODS: The study material consisted of sera from psoriasis patients (n = 80) in active phase and in the remission phase and healthy individuals (n = 80) (controls). Concentrations of AGE-peptides were measured spectrofluorimetrically. Anti-CML and anti-CEL antibody concentrations were determined using ELISA. RESULTS: In psoriasis patients in active phase disease concentrations of AGE-peptides and anti-CML and anti-CEL antibodies in all tested classes were significantly higher than those in healthy individuals. At remission, concentrations of AGE-peptides and tested antibodies decreased significantly, but concentrations of anti-CEL IgG and anti-CML IgG and IgM antibodies were remaining significantly higher in comparison with the control group. CONCLUSION: The obtained results indicate for existence of increased oxidative stress in psoriasis and resulting increased protein glycation and stimulation of the immune system to response to these end-products. Increased protein glyco-oxidation consist a linker between psoriasis and increased prevalence of atherosclerosis, cardiovascular incidents and vascular complications of diabetes.

Administration of low doses of tumor necrosis factor-alpha protects rat liver from ischaemic damage and reperfusion injury.

Liver ischaemia and reperfusion (IR) injury is a significant clinical problem. The aim of our study was to investigate the protective effect of tumor necrosis factor-alpha (TNF-alpha) on rat liver ischaemia-reperfusion injury. A TNF-alpha dose of 3 microg/kg body weight was injected into rats that had undergone partial (70%) ischaemia and reperfusion. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), total blood antioxidant level (using the FRAP test), and the concentrations of TNF-alpha, myeloperoxidase (MPO) and malondialdehyde (MDA) in liver homogenates after 1, 6, and 72 hours of reperfusion were measured. It was demonstrated that, rats subjected to IR, the administration of small doses of TNF-alpha significantly reduced ALT and AST activities after 60- minute liver ischaemia and 1 or 6 hour of reperfusion. The strongest reductions in ALT and AST activities were seen after 1 hour of reperfusion (30% and 35%, respectively). Exogenous TNF-alpha reduced the release of this cytokine in all observed periods, with the greatest reduction observed after 1 hour of reperfusion. Decreases in MPO concentration (by 40-45% in all periods of observation), as a marker of hepatic neutrophil infiltration, and in MDA concentration, the end-product of lipid peroxidation (by 55-60% at all time points), accompanied the reduction of TNF-alpha release. The administration of TNF-alpha to the rats after IR did not alter total plasma antioxidant potential, as assayed by the FRAP test, after 1 hour of reperfusion; however, at the later times a marked increase (approximately 40-50%) occurred. We demonstrated that intraperitoneal injections of small doses of TNF-alpha protect rat livers from IR injury. The mechanism of this protection is related to reductions in the release of TNF-alpha during IR after injection of this cytokine, resulting in reductions in oxidative stress and inflammation during the later phase of reperfusion.

Concentration of anti-7-ketocholesterol antibodies in patient sera after heart transplantation.

Oxidized low-density lipoproteins (ox-LDL) have been shown to be strongly atherogenic and immunogenic in experimental animals and in humans. Autoantibodies against ox-LDL and its various epitopes have been detected in sera and in atherosclerotic lesions. LDL oxidation may also lead to generation of oxidized derivatives of cholesterol (eg, 7-ketocholesterol [7-ketoCh]), which are characterized by high levels of cytotoxicity, muta- and carcinogenicity, as well as immunosuppressive effects and probable immunogenic properties. The aim of this study was to quantify autoantibodies to 7-ketoCh in sera from patients after orthotopic heart transplantation (OHT). IgG antibodies were measured in sera from 81 patients after OHT and 73 healthy volunteers using and enzyme-linked immunosorbent assay (ELISA) method. For the antigen we used 7-ketoCh hemisuccinate conjugated to bovine serum albumin. Anti-7-ketoCh antibodies were detected in all examined sera. Their concentrations were significantly higher among OHT patients than the control group, namely, 109.0 +/- 109.7 vs 53.5 +/- 20.0 U/mL (P < .005). Also, we observed a direct relationship between antibody concentration and time after OHT. Our results showed that the low molecular product of LDL oxidation, 7-ketoCh, was immunogenic possibly playing a role in the progression of atherosclerosis among OHT patients.

Plasma neutrophil gelatinase-associated lipocalin (NGAL) correlations with cystatin C, serum creatinine, and glomerular filtration rate in patients after heart and lung transplantation.

OBJECTIVE: Kidney injury represents a major clinical problem in orthotopic heart transplant (OHT) patients which seriously increases the mortality rate. The aim of this work was to evaluate the utility of a new kidney damage marker-neutrophil gelatinase-associated lipocalin (NGAL)-and its correlations with cystatin C, creatinine, and glomerular filtration rate (GFR) among patients after heart or lung transplantation. MATERIALS AND METHODS: We examined serum samples from 71 patients after heart transplantation, 7 patients after lung transplantation, and 20 healthy controls to measure serum NGAL using an enzyme-linked immunosorbent assay (ELISA) method; cystatin C using latex immunonephelometry; and creatinine using the alkaline-picrate method. RESULTS: Serum NGAL levels were significantly elevated among transplant patients, but did not significantly correlate with cystatin C in the transplant group (R = .13; P = .11) or in the control group (R = .26; P = .26), or GFR in the transplant group (R = -.09; P = .25) or in the control cohort (R = .22; P = .36). In the transplant group, serum NGAL positively correlated with serum creatinine (R = .27; P = .001). CONCLUSIONS: Plasma NGAL was not a specific biomarker for monitoring chronic renal disorders. We did not exclude other pathologies that might contribute to increased serum NGAL levels.

Role of nitric oxide in physiology and pathology of the gastrointestinal tract.

In this paper the physiological role of NO and isoforms of NOS in the gastrointestinal tract and the involvement of NO in pathological processes of digestive tract as well as the perspective of therapeutic use of NO-donating drugs and selective inhibitors of phosphodiesterase in the treatment of gastric diseases were presented.

Serologic markers of celiac disease in psoriatic patients.

BACKGROUND: Aetiopathogenesis of psoriasis is complex and not yet well known. In recent years, it has been observed that psoriasis can coexist with clinically asymptomatic celiac disease and a gluten-free diet helps to obtain remission, even in patients with very chronic lesions. OBJECTIVE: The aim of our work was to investigate how often the positive titres of antibodies characteristic for celiac disease occur in psoriatics' serum in exacerbation in comparison with controls. PATIENTS/METHODS: Serum samples from 67 patients with intensified psoriatic lesions were investigated. Serum from healthy people at a comparable age and with no familial predisposition to psoriasis and celiac disease was the control material. Antibodies against human tissue transglutaminase (recombinant antigen), against tissue transglutaminase isolated from guinea pig's liver and against gliadin were determined by enzyme-linked immunosorbent assay technique. Anti-endomysial antibodies were determined by indirect immunofluorescence method. RESULTS: Patients with psoriasis have significantly higher mean concentrations of antibodies against tissue transglutaminase (human recombinant and guinea pig-derived antigen) and against gliadin for IgA. IgA antibodies against tissue transglutaminase (both antigens) and gliadin positively correlate with psoriasis activity. No anti-endomysial antibodies for IgA were found in any serum. CONCLUSIONS: Our results seem to imply an association between psoriasis and asymptomatic celiac disease/gluten intolerance. High percentage of positive results to guinea pig-derived tTG could be due to cellular activity of tissue transglutaminase in psoriasis.

Anti-ox-LDL and anticardiolipin autoantibodies in patients after cardiac transplantation.

INTRODUCTION: Among cardiac transplantation (OHT) of coronary arterial disease, the pathogenesis can be associated with autoimmunologic effects due to oxidative lipoprotein modification and their change in antigenicity. These factors may lead to lipoprotein vascular changes observed in antiphospholipid syndrome or systemic lupus erythematosus. The aim of the presented study was to evaluate anticardiolipin autoantibodies (ACA) and anti-ox-LDL (antibodies against oxidized LDL) levels in the plasma immunoglobulin IgG class. MATERIALS AND METHODS: The study group consisted of 90 OHT patients (79 men/11 women, age 44 +/- 13); 35 individuals were chosen as a control group. Blood samples were taken twice, in years 2002 and 2004. The investigated group was divided into 2 smaller groups: OHT patients in 2002 and OHT patients before 2002. RESULTS: OHT patients showed significantly higher ACA concentrations compared with the control group (3.53 vs 1.10 GPL U/mL), whereas anti-ox-LDL levels did not differ considerably (494 vs 385 mU/mL). During long-term observation the significant increase in ACA concentration was observed in both patients groups. Significant differences between the 2 OHT patient groups regarding anti-ox-LDL concentration were demonstrated among samples taken in 2002. CONCLUSIONS: The results may indicate the possible role of autoimmunological processes in OHT development. It is necessary to focus further research on the possibilities of developing secondary antiphospholipid syndrome.

Antibacterial effect of autologous platelet gel enriched with growth factors and other active substances: an in vitro study.

Platelet-rich plasma is a new inductive therapy which is being increasingly used for the treatment of the complications of bone healing, such as infection and nonunion. The activator for platelet-rich plasma is a mixture of thrombin and calcium chloride which produces a platelet-rich gel. We analysed the antibacterial effect of platelet-rich gel in vitro by using the platelet-rich plasma samples of 20 volunteers. In vitro laboratory susceptibility to platelet-rich gel was determined by the Kirby-Bauer disc-diffusion method. Baseline antimicrobial activity was assessed by measuring the zones of inhibition on agar plates coated with selected bacterial strains. Zones of inhibition produced by platelet-rich gel ranged between 6 mm and 24 mm (mean 9.83 mm) in diameter. Platelet-rich gel inhibited the growth of Staphylococcus aureus and was also active against Escherichia coli. There was no activity against Klebsiella pneumoniae, Enterococcus faecalis, and Pseudomonas aeruginosa. Moreover, platelet-rich gel seemed to induce the in vitro growth of Ps. aeruginosa, suggesting that it may cause an exacerbation of infections with this organism. We believe that a combination of the inductive and antimicrobial properties of platelet-rich gel can improve the treatment of infected delayed healing and nonunion.

Homocysteine, vitamin B12, and folic acid in age-related macular degeneration.

PURPOSE: Hyperhomocysteinemia is considered an independent risk factor for atherosclerosis and thrombosis. The purpose of this study was to evaluate plasma homocysteine, red blood cell folate, plasma folate, and plasma vitamin B12 concentration in patients with exudative age-related macular degeneration (ARMD). METHODS: The participants of this study included 30 patients aged 60 to 71 years (mean age 66.2+/-3.6) with exudative ARMD. Plasma homocysteine levels were determined by high performance liquid chromatography (HPLC). Red blood cell folate, plasma folate, and plasma vitamin B12 concentration were determined using a standard kit (Dualcount Solid Phase No Boill radioassay kit for B12/folic acid, DPC Diagnostic, USA) by radioassay method. RESULTS: The plasma concentration of Hcy (14.88+/-6.23 micromol/L) in ARMD patients was significantly increased (p<0.0001) compared with the control group (8.72+/-3.34 micromol/L). We found not a significant decrease of the plasma vitamin B12 concentration in the ARMD group (476.88+/-220.91 pg/mL) compared with the control group (527.08+/-208.97 pg/mL). Red blood cell folate (158.44+/-56.30 ng/mL) and plasma folate (6.5+/-3.4 ng/mL) in ARMD patients were also not significantly decreased when compared with the control group (183.86+/-59.33 ng/mL and 7.93+/-5.05 ng/mL). CONCLUSIONS: Hyperhomocysteinemia might be one of the risk factors for the exudative form of ARMD.

Changes in lipid metabolism in women with age-related macular degeneration.

Age-related macular degeneration (AMD) is one of the leading causes of visual loss among people aged 65 and older. At present the origin of AMD still remains unknown. The objective was to evaluate the chosen lipid and lipoprotein concentrations in blood of patients with AMD. Sixty women aged 55-71 (mean age 65.1+/-5.7) were treated in the outpatient ophthalmological clinic for more than two years because of AMD. We evaluated total serum cholesterol (TCH), triglycerides (TG), HDL-cholesterol (HDL), LDL-cholesterol (LDL), lipoprotein (a) (Lp(a)), apolipoprotein AI (Apo AI) and apolipoprotein B (Apo B) by direct spectrophotometry (Human and Randox standard kits, USA). We found a significant increase of TCH, LDL and TG (224.36+/-41.67 mg/dl, 159.02+/-39.66 mg/dl and 120.92+/-42.64 mg/dl), and a significant decrease of HDL (38.68+/-6.36 mg/dl) in the AMD patients when compared with the control group. We have not found a significant difference in the average TG level between the studied groups. The concentration of Apo B was markedly increased (164.66+/-46.46 mg/dl) and Apo AI concentration was markedly decreased (128.9+/-17.01 mg/dl) in the AMD patients when compared with the control group. There was no significant difference in the concentration of the Lp(a) between the two groups. The results of our present study could point to the fact that changes in the lipid metabolism could be one of the very important risk factors involved in the pathogenesis of AMD.

Homocysteine, vitamin B12, and folic acid in age-related macular degeneration.

PURPOSE: Hyperhomocysteinemia is considered an independent risk factor for atherosclerosis and thrombosis. The purpose of this study was to evaluate plasma homocysteine, red blood cell folate, plasma folate, and plasma vitamin B12 concentration in patients with exudative age-related macular degeneration (ARMD). METHODS: The participants of this study included 30 patients aged 60 to 71years (mean age 66.23.6) with exudative ARMD. Plasma homocysteine levels were determined by high performance liquid chromatography (HPLC). Red blood cell folate, plasma folate, and plasma vitamin B12 concentration were determined using a standard kit (Dualcount Solid Phase No Boill radioassay kit for B12/folic acid, DPC Diagnostic, USA) by radioassay method. RESULTS: The plasma concentration of Hcy (14.886.23 micronmol/L) in ARMD patients was significantly increased (p<0.0001) compared with the control group (8,.723,.34 micronmol/L). We found not a significant decrease of the plasma vitamin B12 concentration in the ARMD group (476.88220.91 pg/mL) compared with the control group (527.08208.97 pg/mL). Red blood cell folate (158.4456.30 ng/mL) and plasma folate (6.53.4 ng/mL) in ARMD patients were also not significantly decreased when compared with the control group (183.8659.33 ng/mL and 7.935.05 ng/mL). CONCLUSIONS: Hyperhomocysteinemia might be one of the risk factors for the exudative form of ARMD.

Changes in blood antioxidants and several lipid peroxidation products in women with age-related macular degeneration.

PURPOSE: The aim of this study was to evaluate the ferric reducing ability of plasma (FRAP), selected enzymatic and non-enzymatic components of the antioxidative system, and the intensity of peroxidative processes in the blood of patients with age-related macular degeneration (AMD). METHODS: In the peripheral blood, we evaluated FRAP; concentrations of vitamins C, A, and E; and of thiols. We assayed the activity of enzymatic components of the antioxidative system-superoxide dismutase, catalase, ceruloplasmin and the concentration of reduced glutathione as an indicator of glutathione peroxidase activity. In order to determine the intensity of lipid peroxidation, we measured the concentrations of malondialdehyde and hydroxyalkenales (MDA-HNA) and conjugated diens (CD). RESULTS: We found a significant increase in FRAP in patients with AMD compared with the control group. The average concentrations of vitamins A and C were low and vitamins E and GSH were significantly higher in AMD than in the control group. The activity of almost all the antioxidative enzymes was high. We found a significant increase in MDA-HNA but no difference in CD. CONCLUSIONS: The significantly higher concentration of lipid peroxidation products in patients with AMD indicates an important pathogenic role of oxido-reduction disturbance. The high FRAP concentration may be one of the protective mechanisms in oxidation stress. The adaptive increase of the antioxidant barrier mostly involves the enzymatic components.

[Plasma oxysterols and vitamin E concentrations and lipid profile in morbidly obese women]. / Stezenie oksycholesteroli i witaminy E a profil lipidowy osocza u kobiet ze skrajna otyloscia.

Morbid obesity (BMI > or = 40 kg/m2) is accompanied by lipid disturbances which may be involved in the increased incidence of atherosclerosis, arterial hypertension and non-insulin-dependent diabetes mellitus. The aim of the study was to assess concentrations of total cholesterol (TC), HDL-cholesterol, LDL-cholesterol, triglycerides (TG), products of cholesterol peroxidation--oxysterols, and the major lipophilic antioxidant--vitamin E, in morbidly obese women without coexisting diseases. The study was performed in 11 morbidly obese women (BMI 42.21 +/- 2.21 kg/m2) and 11 healthy volunteers (BMI 23.0 +/- 2.31 kg/m2). Obese women demonstrated higher concentrations of TG (2.03 +/- 0.78 vs. 0.99 +/- 0.37 mmol/l; p < 0.05), 7-ketocholesterol (7-K) (89.85 +/- 63.03 vs. 41.90 +/- 17.33 ng/ml; p < 0.05) and 7-hydroxycholesterol (7-OH) (456.04 +/- 199.22 vs. 132.37 +/- 53.96 ng/ml; p < 0.05), and lower HDL-cholesterol level (0.74 +/- 0.10 vs. 1.30 +/- +/- 0.17 mmol/l; p < 0.05) compared to the control group, while there were no significant differences between the two groups in concentrations of TC, LDL-cholesterol and vitamin E. Plasma vitamin E/(TC + TG) ratio was lower in obese women (6.42 +/- 2.61 vs. 10.76 +/- 4.57 mumol/mmol; p < 0.05). Tocoferols concentration correlated positively with TG (r = 0.45; p < 0.05) and negatively with 7-OH (r = -0.44; p < 0.05) levels. Moreover, concentration of 7-K correlated positively with the level of HDL (r = 0.54; p < 0.05). In conclusion, despite normal TC and LDL-cholesterol concentrations, there are disturbances in cholesterol peroxidation processes, with the rise in oxysterol levels and the decrease in vitamin E concentration in lipoproteins, which may be involved in the increased incidence of cardiovascular diseases in morbidly obese women.

Application of liquid separation techniques to the determination of the main urinary nicotine metabolites.

A rapid procedure for the analysis of the main nicotine metabolites (cotinine, trans-3'-hydroxycotinine) in urine has been worked out. The procedure includes isolation of nicotine and its metabolites from urine by means solid-liquid extraction technique using resin Amberlite XAD-2 and then quantitation by the use of thin-layer chromatography with densitometry (in reflection mode). GC-MS was applied to confirm the results obtained by TLC. The procedure was applied to the analysis of cotinine concentrations in urine samples taken from children living in Upper Silesia region (Poland). Among 444 investigated children we did not find cotinine almost in 60% but in 15% of this population, there were children who could have been exposed to cigarette smoke.

[The estimation of L-ascorbic acid serum concentration and selected parameters of metabolism of serum lipids in pregnant women with diabetes]. / Ocena stezen kwasu L-askorbinowego oraz wybranych parametrów gospodarki lipidowej we krwi kobiet ciezarnych z cukrzyca.

Diabetes during pregnancy is a pathological state causing many metabolic disorders, not only in carbohydrate metabolism. The aim of the study was to estimate the changes of lipids metabolism and changes of L-ascorbic acid serum concentration in pregnancy complicated by diabetes. In tested groups values of lipids metabolism parameters were lower in diabetic group compared to control group. The lowest mean values of concentration of total cholesterol, LDL cholesterol and triglycerides were found in gestational diabetes group, while the lowest mean value of HDL cholesterol concentration was found in pregestational diabetic group. The L-ascorbic acid mean values correlated negatively with total cholesterol concentrations, LDL cholesterol and triglycerides levels.