Usefulness of New Indicators of Obesity (BAI and VAI) in Estimation of Weight Reduction.

Objective: Obesity is responsible for a large proportion of the total burden of diseases in Europe. The aim of this study was to compare classic anthropometric parameters with new obesity-related indices (the visceral adiposity index [VAI] and the body adiposity index [BAI]) and to examine their usefulness in the assessment of weight reduction and its effect on the leptin concentration.Methods: We tested 50 patients with obesity, undergoing the implementation of the BioEnterics® Intragastric Balloon (BIB-system) for 6 months. The leptin concentrations and the calculated subcutaneous fat thickness as well as the anthropometric indices were determined prior to the implementation and after the balloon removal.Results: The therapy resulted in a drop in the BAI value without a significant effect on the VAI in subgroups of patients with class I, II, and III obesity. The therapy-induced changes in BAI both in the entire tested group and in patients with class I and II obesity correlated well with the percentage of drop in body weight, body mass index (BMI), subcutaneous fat thickness, waist-to-height ratio (WHtR), and leptin concentration. Changes in the VAI value correlated with changes in the body weight, BMI, WHtR values, subcutaneous fat thickness, and leptin concentration decrease only in patients with class III obesity.Conclusions: BAI seems to be a better indicator than VAI for reflecting the reduction in body weight in people with class I and II obesity treated with the BIB-System. VAI appears to be a better obesity indicator than BAI only in people with class III obesity.

The Effect of Noninvasive Bariatric Surgery on the Levels of Certain Adipokines and Atherosclerosis Risk Factors in Patients with Metabolic Syndrome.

Objective: Besides glucose intolerance, dyslipidemia, and hypertension, visceral obesity is one of the most important atherogenic pathological factors in patients with metabolic syndrome (MetS). The aim of this study is to examine whether weight loss following BioEnterics Intragastric Balloon (BIB-system) therapy affects adipokine concentration and atherosclerosis risk factor profile in patients with MetS.Methods: The study group comprised 30 patients (17 female, BMI = 38.5 ± 8.6 kg/m2; 13 male, BMI = 43.3 ± 7 kg/m2) with MetS qualified to BIB-system therapy. The control group included 18 age matched healthy volunteers (10 female, BMI = 23.3 ± 2.8 kg/m2 and eight male, BMI = 27.3 ± 0.9 kg/m2). Biochemical analyses of blood samples and anthropometric measurements were conducted, before and after six-month BIB system therapy.Results: BIB therapy resulted in a significant drop in body weight, and body fat percentage, and in BMI, VAI, WHtR, BAI, TG, glucose, hsCRP, and leptin levels. In addition Tc/HDL, LDL/HDL, TG/HDL, and leptin/adiponectin ratios fell significantly, and adiponectin concentration increased. All anthropometric parameters apart from Tc and hsCRP, were significantly different post-therapy compared to healthy controls. The therapy induced downregulation of hsCRP which was positively correlated with the reduction in body weight, BMI and BAI. The decrease in leptin concentration correlated positively with the fall in total cholesterol and body weight. The fall in leptin/adiponectin ratio positively correlated with the downregulation of BAI and body fat.Conclusion: BIB therapy appears to have beneficial effects on MetS. This is indicated by amelioration of the pro-inflammatory status related to obesity, demonstrated by an improved lipid profile significant downregulation of hsCRP concentration following therapy.

Fibrinogen concentrations in ischaemic stroke patients with metabolic disorders.

BACKGROUND: Hyperfibrinogenemia plays a crucial role in the coagulation cascade leading to the formation of clots. It is involved in the process of platelet aggregation, primary haemostasis, and leukocyte-endothelial cell interactions. The aim of our study was to assess the correlations between fibrinogen concentration and particular risk factors for vascular diseases and atherosclerotic changes in stroke patients. METHODS: The study group consisted with 94 patients with acute ischaemic stroke with normo- or hyperglycaemia and normoor hyperlipidemia. 21 healthy subjects served as a control group. Fibrinogen level, HbA1c, and lipid profile were measured in all patients. Using a flow cytometer, we assessed CD61 positive microparticles which were defined as platelet-derived microparticles (PDMPs). The level of sP-selectin in serum was measured using the ELISA method. RESULTS: A significant positive correlation was observed between fibrinogen concentration and sP-selectin (p = 0.001), HbA1c (p < 0.05) level, and percentage of PDMPs (p < 0.05) in the study patients. Furthermore, we noticed a significant negative correlation between fibrinogen concentration and the level of HDL (p < 0.05). No correlation was observed between fibrinogen and TC, LDL and TG levels. CONCLUSIONS AND CLINICAL IMPLICATIONS: Our findings suggest that an elevated fibrinogen level may represent a marker of prothrombotic condition exacerbated in the state of hyperglycaemia and activation of platelets and endothelial cells. This suggests an important role played by fibrinogen in the process of thrombogenesis.

Impact of Allergic Rhinitis on Nasal Mucociliary Clearance Time in Children.

BACKGROUND: Mucociliary clearance is one of the most important protective functions of the airway. Previous studies, checking the influence of allergic rhinitis (AR) on mucociliary clearance time (MCT), were made on small patient groups and brought contradictive results. OBJECTIVES: The aim of the study is to confirm whether AR in children influences MCT. METHODS: The examined group consisted of 842 AR children. A total of 96 children with no history of allergy rhinitis served as a comparative group. All patients underwent saccharin and skin prick tests and tests for blood eosinophilia, nasal eosinophilia, vitamin D3 serum concentration, total and specific IgE serum concentration. RESULTS: Nasal MCT was significantly longer in AR patients (mean ± SD: 10.5 ± 5.65 min) compared to controls (mean ± SD: 7.25 ± 4.3 min). Percentage of eosinophils in nasal smears in patients was significantly higher compared to controls and a weak, but significant positive correlation was observed between the percentage and MCT (r > 0.10, p < 0.008). Patients with intermittent and persistent moderate/ severe AR had significantly longer MCT and higher eosinophilia in nasal smears compared both to patients with intermittent and persistent mild RA and controls. No correlation was observed between MCT and: participant's age, total serum IgE, vitamin D3 serum concentration, absolute number or percentage of eosinophils in blood, prick test results or duration of illness. CONCLUSIONS: AR affects the mucociliary clearance in children, and its deterioration is related to more severe rhinitis with higher intensity of local nasal inflammation, reflected in nasal smear eosinophilia.

sP- and sE-selectin in stroke patients with metabolic disorders.

BACKGROUND: Activation of platelets and endothelial cells plays an important role in the pathogenesis of atherosclerosis and thrombotic disorders. The aim of our study was to assess the relationship between the metabolic disorders and markers of platelet activity and vascular injury in patient with acute ischemic stroke. MATERIAL AND METHODS: The study group consisted of 84 patients with acute non-lacunar ischemic stroke divided into four subgroups with: (1) normolipidemia and normoglycemia, (2) normolipidemia and hyperglycemia, (3) hyperlipidemia and normoglycemia, (4) hyperlipidemia and hyperglycemia. 21 healthy subjects served as controls. We analyzed the concentration of adhesion molecules sP-selectin and sE-selectin in serum collected from all studied groups using ELISA method. RESULTS: We observed significantly higher sE- and sP-selectin concentration in patients with hyperglycemia and hyperlipidemia compared to both control subjects and patients with normolipidemia and normoglycemia. We did not observe additional effect of comorbid hyperlipidemia and hyperglycemia on studied markers. Soluble E- and P-selectin concentration correlated positively with LDL, TC and HbA1c level in all stroke patients. CONCLUSION: Soluble E- and P-selectin, blood markers of vascular injury and platelet activation, could be useful in the assessment of atherothrombotic properties of hyperglycemia and hyperlipidemia in stroke patients.

Basic inflammatory indices and chosen neutrophil receptors expression in chronic haemodialysed patients.

AIM OF THE STUDY: End stage renal disease (ESRD) patients on chronic haemodialysis (HD) are immuno-compromised and prone to infection. Toll-like receptors (TLRs) play a role as both primary sensors of pathogen invasion and activators of inflammatory reaction. To test if the immune impairment in HD patients is connected with the defective expression of the neutrophil TLRs, we aimed to examine their expression and chosen inflammatory indices. MATERIAL AND METHODS: We tested CD14, TLR4, and TLR9 expressions on neutrophils using flow cytometry. Soluble CD14, C-reactive protein (CRP), and mannose-binding lectin (MBL) concentrations were tested using the ELISA method in 31 ESRD patients on chronic haemodialysis programs and in 17 healthy control subjects. RESULTS: Neutrophil TLR4 and TLR9 expressions did not differ significantly compared to the controls. The ESRD patients had markedly increased CRP and sCD14 levels alongside decreased MBL concentrations and neutrophil CD14 expression. The TLR4 expression correlated well with both TLR9 and CD14 neutrophil expressions; however, the increased CRP in the blood did not correlate with the MBL concentration or TLR expression. CONCLUSIONS: The chronic program of haemodialysis and biochemical disorders in ESRD patients result in a low-grade chronic inflammation with no significant impact on the expression of neutrophil TLR4 and TLR9.

[Chosen anthropometric parameters and concentrations of leptin and adiponectin in extreme obese patients treated with implantation of a gastric balloon]. / Wybrane parametry antropometryczne oraz stezenie leptyny i adiponektyny u pacjentów ze skrajna otyloscia leczonych implantacja balonem zoladkowym.

UNLABELLED: Fatty tissue derived adipocytokines regulate appetite, but in abnormal concentration impair systemic metabolic homeostasis and make the patients prone to inflammatory related disorders. The aim of study was to examine whether weight loss in patients after implementation of a gastric balloon is reflected in changes in chosen anthropometrical parameters and in the concentration of leptin and adiponectin in serum. MATERIALS AND METHODS: The study group consisted of 18 extreme obese patients (BMI>39.9, mean age 39.5±12.1 years, 12 men, 6 women), undergoing implementation of a gastric balloon for 6 months. The control group consisted of 18 healthy volunteers. The adiponectin and leptin concentrations in the sera and the calculated % body fat and indicators: BMI, WHR, VAI, BAI, WHtR were determined prior to implementation and after the balloon removal and then further parameters were calculated: % excessive weight loss, % weight loss. RESULTS AND CONCLUSIONS: All the parameters and leptin concentration in the tested group were markedly upregulated and adiponectin concentration was significantly lower compared to controls. Reduction in the body mass in patients subjected to BIB, reflected in leptin and anthropometrical parameters down-regulation, (except WHR and VAI), was accompanied with normalization of adiponectin concentration that affect metabolism and is important regulator of hunger and satiety.

The influence of uremic high cystatin C concentration on neutrophil apoptosis and selected neutrophil functions isolated from healthy subjects.

BACKGROUND: Cystatin C (cC) is a cysteine protease inhibitor that may influence immune response. Our aim was to test the effect of a high concentration of cC, characteristic for uremic patients, on neutrophil (PMN) apoptosis and respiratory burst, as well as the cC secretion from PMNs stimulated with proinflammatory cytokines. MATERIAL/METHODS: PMNs from 35 healthy volunteers aged 27-61 years were cultured in presence of cC, IL-1beta or TNF-alpha. The percentage of apoptotic cells based on DNA depletion, Fas, FasL and caspase -3 expression were assessed. CC concentrations were determined by ELISA test. The influence of cC on spontaneous, fMLP-, PMA- or OZ-induced burst response of PMNs was tested using chemiluminescence. RESULTS: PMN cultured in the presence of cC resulted in a significant drop in apoptotic cell percentage (38% [11%; 65%]) compared both to control (70% [29%; 92%], and to the cells cultured with TNF-alpha (58% [24%; 85%]). These differences were not accompanied by Fas, FasL and caspase-3 expression changes. Spontaneous, fMLP- and PMA-stimulated oxidative burst of PMNs preincubated with cC were significantly downregulated. IL-1beta markedly diminished and TNF-alpha significantly increased cC concentration in culture supernatants. CONCLUSIONS: The presented results suggest that antiapoptotic activity of cC results from its inhibitory effect on ROS production. Thus, the higher concentration of cC characteristic for uremic patients may modulate acute inflammation through maintaining PMN longevity and inhibiting their respiratory burst and proinflammatory cytokine-related changes in cC release from PMNs.

The influence of hyperlipidemia on platelet activity markers in patients after ischemic stroke.

BACKGROUND: To investigate the relationship between hyperlipidemia and platelet activation markers--platelet and soluble P-selectin (sP-selectin), and platelet-derived microparticles (PDMPs)--in patients after ischemic stroke. METHODS: 41 patients after ischemic stroke (>3 months) confirmed by CT were divided into 2 groups: with hyperlipidemia (HL, n = 21) and normolipidemia (NL, n = 20). Twenty healthy subjects served as controls. CD62P-positive platelets and PDMPs in whole blood were analyzed by the use of a flow cytometer and anti-CD61 and anti-CD62P monoclonal antibodies. Platelets were activated by thrombin (0.08 units). The level of sP-selectin in serum was measured by ELISA. RESULTS: We observed a significantly higher CD62P expression and percentage of CD62P-positive resting and thrombin-activated platelets in the HL as compared to the NL group. The sP-selectin concentration was also significantly higher in HL than NL subjects (p < 0.05). Moreover, we observed a significantly higher percentage of PDMPs in patients after stroke (NL: p < 0.05; HL: p = 0.005) in comparison with the control group. CONCLUSIONS: Patients after stroke present symptoms of platelet hyperreactivity. HL in the patients may be a risk factor for vascular events due to the increase in platelet activation.

Effect of luteinizing hormone-releasing hormone (LHRH) analogue treatment on a cytokine profile in prostate cancer patients.

The aim of the study was to test serum concentrations of the chosen cytokines in patients with prostate cancer (PCa) treated with an luteinizing hormone-releasing hormone (LHRH) analogue. We tested interleukin (IL)-2, IL-10, tumor necrosis factor (TNF)-alpha, interferon (INF)-gamma in blood at three time points; I - before the injection, II - 10 days and III - 20 days after the injection in 14 men with PCa. Patients had one depot injection of the LHRH analogue monthly. The cytokine concentrations in serum samples were determined by ELISA method. Prostate specific antigen (PSA) level was examined before and after six months of the LHRH analogue treatment. After six months of the therapy, we observed normalization of serum PSA value from 16.48 ng/ml to 1.45 ng/ml. LHRH analogue injection resulted in a significant drop of the IL-2 concentration, and the value gradually returned to normal in the next 20 days. IL-10 concentration transiently increased and then was down-regulated. Serum TNF-alpha and INF-gamma concentrations in PCa patients were significantly lower compared to controls and were not affected by the treatment. LHRH analogue treatment in PCa patients modulates concentrations of the chosen cytokines which may result both in antitumor and a transient immunosuppressive effect.

[Estimation of relation between homocysteine concentration and selected lipid parameters and adhesion molecules concentration in children with atherosclerosis risk factors]. / Ocena zaleznosci pomiedzy stezeniem homocysteiny a stezeniem wybranych parametrów przemiany lipidowej i czasteczek adhezyjnych u dzieci z czynnikami ryzyka miazdzycy.

UNLABELLED: Atherosclerosis begins in childhood. At present among numerous risk factors of atherosclerosis the role of hiperhomocysteinemia in development of cardiovascular heart disease is taken under consideration. Atherogenic effect of homocystein is related to its cytotoxin action, conducting to endothelial dysfunction and damage. It is correlated with increase of the lipid levels in the blood serum and change of expression of the soluble forms of adhesion molecules. The aim of this study was to estimate relations between the homocystein serum concentration, expression of the selected adhesion molecules and the lipid levels in the blood serum in children with atherosclerosis risk factors. MATERIAL AND METHODS: The group consisted of 670 children, 76 of them had atherosclerosis risk factors. In further examination 48 children have taken a part, whose parents were agreed for theirs participation in the program. The comparative group composed of 25 children without the risk factors. We determined total cholesterol (TC), triglycerides (TG), LDL cholesterol fraction (LDL-C), HDL cholesterol fraction (HDL-C), serum homocysteine concentration (Hcy), the expression of the soluble forms of adhesion molecules (sCAM): sP-selectin and sVCAM-1 (vascular cell adhesion molecule-1). RESULTS: Obesity, hypertension and lipid disorders in the shape of higher concentration of TC, LDL-C, TG and lower HDL-C were the most frequent risk factors in the investigated children. No significant differences in serum homocysteine concentration were observed between the investigated groups. However, its concentration was significantly higher in children with two atherosclerosis risk factors. No significant differences in expression of s-VCAM-1 were observed in the investigated groups, concentration of sP-selectin was significantly higher in children with atherosclerosis risk factors (p<0.05). Statistically significantly higher serum concentrations of lipid levels were found in children with hyperhomocysteinemia. CONCLUSION: Higher concentration of the serum homocysteine and chosen adhesion molecules in children with atherosclerosis risk factors might potentially constitute the marker of early atherosclerotical risk development.

[Estimation of selected parameters antioxidant barrier and lipid parameters in children with atherosclerosis risk factors]. / Ocena wybranych parametrów bariery antyoksydacyjnej i przemiany lipidowej u dzieci ze stwierdzonymi czynnikami ryzyka miazdzycy.

UNLABELLED: Atherosclerosis is one of the most common causes of mortality in developed countries. One of the many factors in its etiopathogenesis are reactive oxygen species (ROS), excess of which comes from disturbances of enzymatic and nonenzymatic mechanisms of antioxidant barrier. THE AIM OF THIS STUDY was to estimate relations between the activity of chosen parameters antioxidant barrier and the lipid levels in children with atherosclerosis risk factors. MATERIAL AND METHODS: The study was carried out on 48 children with atherosclerosis risk factors. The control group consisted of 25 healthy children. Total cholesterol (TC), Triglycerides (TG), HDL-C, LDL-C were determined by enzymatic method. The activity of superoxide dismutase (SOD) was estimated by Misra and Fridivich's method, glutathione peroxidase (GSH-Px) with method proposed by Sedlak and Lindsay in modyfication Little and Brien and the activity o catalase (CAT) by Beer and Sizer's method. Malonyl dialdehyde (MDA) concentration was determined by Placer's method. RESULTS: Obesity, lipid disorders, and hypertension were the most frequent risk factors in the investigated children. Statistically significant higher concentration of TC, LDL-C, TG and lower HDL-C were observed in children with atherosclerosis risk factors. Activity of glutathion peroxidase was statistically significant higher in children with atherosclerosis risk factors (p < 0.001), activity of catalase was lower (p < 0.05). No significant differences in activity of superoxide dismutase and malonyl dialdehyde were observed in the investigated groups. CONCLUSION: Modificated activity of selected antioxidants in children with atherosclerosis risk factors could determine an index of antioxidant barrier and might potencially constitute the marker of early atherosclerotical risk development.

Serum matrix metalloproteinases MMP-2 and MMP-9 and metalloproteinase tissue inhibitors TIMP-1 and TIMP-2 in diabetic nephropathy.

BACKGROUND: The regulation of mesangial extracellular matrix (ECM) turnover engages a number of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). High glucose concentration affects ECM degradation and the activities of MMPs and TIMPs. ECM accumulation is involved in the pathogenesis of diabetic nephropathy. METHODS: Serum MMP-9, MMP-2, TIMP-2 and TIMP-1 were measured with ELISA in patients with either chronic renal failure (CRF, n=20), type 2 diabetes mellitus (DM2, n=16) or diabetic nephropathy (DM2+CRF, n=14), and healthy controls (n=20). RESULTS: Diabetic nephropathy was related with profound decrease of serum TIMP-2 (122.2 +/- 47.2 vs. 263.0 +/- 89.2 ng/mL), TIMP-1 (242.5 +/- 96.9 vs. 347.4 +/- 87.2 ng/mL) and MMP-2 (385.4 +/- 42.6 vs. 517.2 +/- 75.4 ng/mL) (p<0.001). Both TIMP-1 and TIMP-2 were reduced in diabetic nephropathy in comparison with either diabetes alone (p<0.01 and p<0.001; respectively) or CRF alone (p<0.001 for both). An approximately 2-fold increase of MMP-9/TIMP-1 and MMP-2/TIMP-2 ratio was found in diabetic nephropathy when compared with diabetes with normal renal function (p<0.01). Further, in DM2 patients, TIMP-2 was decreased when compared with CRF alone (219.2 +/- 71.8 vs. 296.8 +/- 58.4 ng/mL). MMP-2 was lowered in both groups of DM2 and CRF patients (413.8 +/- 59.0 ng/mL and 409.7 +/- 93.1 ng/mL, vs. normal control value of 517.2 +/- 75.4 ng/mL; p<0.001). CONCLUSIONS: These data indicate that circulating TIMP-1, TIMP-2 and MMP-2 are decreased in patients with diabetic nephropathy when compared with either CRF or diabetes.

[Effect of the acetylosalicyd acid (ASA) and ticlopidine therapy on clinical condition and parameters of blood platelets in patients with peripheral arterial occlusive disease (PAOD)]. / Ocena wplywu terapii kwasem acetylosalicylowym i tiklopidyna na stan kliniczny oraz parametry plytek krwi u chorych na miazdzyce zarostowa konczyn dolnych (arteriosclerosis obliterans).

UNLABELLED: Hyperreactivity of platelets is one of the agents promoting atherosclerosis and its organ complications THE AIM: To assess antiplatelet therapy with acetylsalicylic acid (ASA) and ticlopidine on clinical condition and chosen parameters of blood platelet activity and reactivity in patients with PAOD. MATERIAL AND METHODS: Twenty eight patients, aged 40-65 years, were enrolled to the study after clinically and echographically established diagnosis. The patients were randomly divided into two groups: 13 were treated with ASA 300 mg daily and 15 with ticlopidine 2 x 250 mg daily. The therapy lasted two months. Clinical stress tests were done on moving track according to thB modified Bruce's protocol and blood platelets activity parameters on flow-cytometer. Percentage of spontaneous platelet aggregates, microplatelets and platelets expressing CD62P, Cd42b and CD41 were tested. RESULTS: Supplementation therapy with ASA or ticlopidine to the standard therapy with pentoxyphylline resulted in a marked increase in the distance of claudication evaluated subjectively and objectively on moving track and revealed silent myocardial ischemia in 4 PAOD patients. In platelet activation tests, both at rest and after the stress test, independently of the antiplatelet agent used, we observed significant drop in the percentage of platelet aggregates and symptoms of vWF receptor activation. No effect of the therapy on CD62P expression, reflecting platelet release reaction and CD41 expression (the fragment of receptor for fibrynogen) were noted. CONCLUSIONS: Both antiplatelet drugs have beneficial effect on clinical condition of PAOD patients and the effect may results from their anti-aggregative and anti-adhesive properties.

[Homocystein serum levels and lipid parameters in children with atherosclerosis risk factors]. / Homocysteina i wybrane parametry przemiany lipidowej u dzieci z czynnikami ryzyka miazdzycy tetnic.

UNLABELLED: Atherosclerosis is a disease of adult patients, however, it begins in childhood and progresses from fatty streaks to raised lesions in arteries in adolescence and young adults. Clinical manifestation of atherosclerosis in adulthood depends on the risk factors such as: lipid disorders, obesity, hypertension, smoking habits and family history of CHD. High serum homocysteine concentration is increasingly recognised as a new risk factor for atherosclerosis and other vascular diseases. Atherogenic effect of homocystein is related to cytotoxin action on the endothelial cells and their function. The aim of this study was to estimate relations between the homocysteine serum concentration and the lipid levels in children with atherosclerosis risk factors. MATERIAL AND METHODS: The study was carried out on 48 children with atherosclerosis risk factors. The control group consisted of 25 healthy childrens. Total cholesterol (TC), Triglycerides (TG), HDL-C, LDL-C were determined by enzymatic method. Concentration of homocysteine was estimated by immunoenzymatic method (ELISA). RESULTS: Obesity, lipid disorders, and hypertension were the most frequent risk factors in the investigated children. Statistically significant higher concentration of TC, LDL-C, TG and lower HDL-C were observed in children with atherosclerosis risk factors. No significant differences in homocystein concentration were observed in the investigated groups, but homocystein concentration was significantly higher in group of children with atherosclerosis risk factors. CONCLUSION: We observed that increased number of the risk factors is followed by high homocystein concentration in the serum.

The impact of hyperglycemia and hyperlipidemia on plasma P-selectin and platelet markers after ischemic stroke.

INTRODUCTION: Platelet activation plays a key role in the pathogenesis of ischemic cerebrovascular diseases. Thus, it is very important to identify novel pharmacological targets for platelet inhibition to improve ischemic stroke treatment. The aim of the study was to assess the relationship between metabolic disorders and platelet activity markers in patients with acute ischemic stroke. MATERIAL AND METHODS: Ninety-four patients with acute ischemic stroke were divided into four groups with: normolipidemia and normoglycemia (NL/NG), n = 25; normolipidemia and hyperglycemia (NL/HG), n = 21; hyperlipidemia and normoglycemia (HL/NG), n = 27; hyperlipidemia and hyperglycemia (NL/NG), n = 21. Twenty-one healthy subjects served as controls. We assessed the CD62P expression on resting and thrombin-activated blood platelets using the flow cytometer and anti-CD61 and anti-CD62P monoclonal antibodies. CD61-positive microparticles were defined as platelet-derived microparticles. The level of sP-selectin in serum was measured by the ELISA method. RESULTS: We observed a significant influence of hyperlipidemia and hyperglycemia on sP-selectin concentration. A strong correlation between higher sP-selectin concentration and enhanced LDL (p = 0.001), total cholesterol (p = 0.02), HbA1c level (p < 0.001) was noticed. The level of sP-selectin and PDMPs (p < 0.001) were significantly higher in all groups of stroke patients compared with the controls. CD62P expression on resting and thrombin activated platelets were significantly lower in groups of patients with stroke. CONCLUSIONS: Hyperlipidemia and hyperglycemia exert an equal stimulatory effect on tested platelet markers but with no synergistic action in stroke patients with both of the metabolic comorbidities. sP-selectin concentration in stroke patients best reflects the impact of hyperglycemia and hyperlipidemia on vascular lesions and platelet activation.

Platelet reactivity and mean platelet volume as risk markers of thrombogenesis in atrial fibrillation.

Atrial fibrillation (AF) is associated with increased risk of thromboembolic complications. One of the markers of the increased risk of hypercoagulable state is platelet hyperreactivity. The aim of the study was to assess impact of arrhythmia on platelet reactivity. METHODS: The study included 36 (mean age 48,3; range 21-60) male patients with lone atrial fibrillation, with exclusion of concomitant diseases known to trigger hypercoagulable state. The AF patients underwent cardioversion to restore sinus rhythm and were subsequently under observation for 1month. Echocardiography, ECG and blood collection was performed before cardioversion (T0) and 4weeks after successful cardioversion (T1). During the study period patients have been contacted and examined every week and 24h ECG monitoring was performed. Platelet reactivity was assessed based on changes of CD62 and CD42b expression on platelet surface after stimulation with thrombin. Also changes in MPV were assessed. RESULTS: In all patients sinus rhythm was maintained at the end of the study period, however in 14 patients recurrences of AF were observed, confirmed by 24h ECG monitoring (atrial fibrillation recurrence group - AFR) and 22 patients maintained sinus rhythm throughout the whole study period (SR group). Mean fluorescence intensity (MFI) of CD62 on thrombin stimulated platelets decreased significantly 4weeks after electrical cardioversion as compared to T0 (48.04±22.42 vs 41.47±16.03; p<0.01). Also MFI of CD42b on thrombin stimulated platelets decreased significantly 4weeks after electrical cardioversion as compared to T0 (22.16±10.82 vs 12.06±5.99; p<0.0001). Platelets reactivity estimated by CD 62 expression in SR group decreased significantly after 4weeks observation (58.01±15.26 vs 46.57±13.44; p<0.001) opposite to AFR group 35.66±21.87 vs 34.54±16.4; p-ns). Moreover there were significant differences between basal reactivity during AF between SR and AFR groups (58.01±15.26 vs 35.66±21.87; p-0.01). MFI of CD42b on thrombin stimulated platelets decreased significantly both in AFR and SR groups (22.05±11.36 vs 13.8±6.03; p<0.001 and 21.87±14.18 vs 10.04±5.09; p<0005). MPV decreased significantly 4weeks after electrical cardioversion as compared to T0 (8.81±0.19 vs 8.42±0.14; p<0.0001). CONCLUSION: The changes of platelet reactivity to thrombin observed after restoration of sinus rhythm in patients prove that arrhythmia intrinsically leads to increased reactivity of platelets.

TNF-alpha priming effect on polymorphonuclear leukocyte reactive oxygen species generation and adhesion molecule expression in hemodialyzed patients.

INTRODUCTION: The study aimed to assess reactive oxygen species generation and the expressions of some surface antigens on polymorphonuclear leukocytes (PMNs) in patients on regular hemodialysis (HD) treatment. MATERIALS AND METHODS: The respiratory burst of PMNs was determined with luminol-dependent chemiluminescence (CL) in resting cells and following N-formyl-methionyl-leucyl-phenylalanine (fMLP), phorbol 12-myristate 13-acetate (PMA), or opsonized zymosan (OZ) stimulation and expressed in arbitrary CL units times assay-time (aU x min). The expressions of CD11b/CD18, CD10, and CD13 receptors were determined with flow cytometry. RESULTS: Basal PMN CL was increased in HD patients to up to 1285 +/- 129 aU x min compared with 895 +/- 88 aU x min in healthy controls (p < 0.05). The CL of unprimed PMNs increased after fMLP stimulation from 3085 +/- 746 to 4529 +/- 808 aU x min, and after OZ stimulation from 12945 +/- 1296 to 14678 +/- 1355 aU x min. PMA-stimulated CL of PMNs was similar to control values. The oxidative burst in PMNs from HD patients and healthy controls was similar in response to TNF-alpha alone. The CL of TNF-alpha-primed PMNs in HD patients was significantly lower than CL measured in healthy controls (p < 0.05). The expressions of CD10 and CD13 metalloproteinase receptors were also increased (p < 0.05). Although CD11b expression was significantly increased at rest and after fMLP stimulation, the expression of another beta-integrin heterodimer compound, CD18, was not increased. CONCLUSIONS: These results provide evidence that TNF-alpha priming of PMNs is down-regulated in HD patients despite constitutive up-regulation of resting cytotoxicity and enhanced expression of adhesion and metalloproteinase receptors.

Quantitative analysis of cyclin E and protein p34 cdc2 expression in superficial bladder cancer.

In the present study, the expression of cyclin E and kinase p34 cdc2 was investigated in preinvasive bladder tumors. The study material consisted of bladder sections (grades: GI--16 cases, GII--10, and GIII--12) collected from 38 patients in the course of the tumor electroresection. Immunohistochemical examinations were carried out with immunoperoxidase method. Antigens were labeled with NCL-CYCLIN E or NCL-p34 cdc2 monoclonal antibodies (Novocastra, UK). Positive reaction was demonstrated using ABC-universal Kit (Novocastra, UK). Differences in the protein expression in relation to the tumor grade were determined with a non-parametric Mann-Whitney's test. Increasing grade of tumors was associated with down regulation of cyclin E visible as lower percentage of cyclin E-positive cells. These changes were statistically significant for GI group as compared to groups GII and GIII (p<0.001). There were no differences between the study groups in the p34 protein expression. Cyclin E expression was inversely correlated with tumor grade therefore may be helpful in establishing therapeutic procedure.

Platelet Reactivity in Patients With Stroke and Hyperlipidemia, GPIbα Assessment.

The aim of this study was to assess platelet reactivity in patients after ischemic stroke and to investigate the influence of hyperlipidemia (HL) on platelet activity markers. A total of 41 patients after ischemic stroke were divided into the following 2 groups: patients with HL and patients with normolipidemia. Expression of CD42b on resting, thrombin-activated blood platelets, and fibrinogen level was assessed. The CD42b-positive platelets were analyzed using the flow cytometer, anti-CD61, and anti-CD42b monoclonal antibodies. The results confirmed increased platelet reactivity to thrombin in all patients after ischemic stroke manifested by significantly lower CD42b expression and percentage of CD42b(+) platelets after activation by thrombin. The influence of HL on the expression of CD42b on resting and thrombin-activated platelets was not found. However, increased level of fibrinogen but no influence of HL on fibrinogen concentration was observed in patients after ischemic stroke. Increased susceptibility to platelet agonists was found in patients after ischemic stroke in the convalescent phase.