Serum metalloproteinases MMP-2, MMP-9 and metalloproteinase tissue inhibitors TIMP-1 and TIMP-2 in patients on hemodialysis.

BACKGROUND: We assessed the effect of hemodialysis (HD) and chronic kidney disease (CKD) on the serum levels of metalloproteinase-2 (MMP-2), MMP-9 and metalloproteinase tissue inhibitors (TIMP-1) and TIMP-2. METHODS: 18 patients on regular HD treatment with low-flux, cuprophane membrane, 15 non-dialyzed patients with CKD and 15 healthy controls were sampled. The serum MMP and TIMP concentrations were determined by ELISA assays. RESULTS: MMP-9, TIMP-1, and TIMP-2 serum levels were significantly decreased in HD patients to 32.7 ± 20.1 ng/ml, 178.8 ± 73.0 ng/ml, and 103.4 ± 55.3 ng/ml compared with 482.3 ± 139.5, 367.6 ± 75.5 ng/ml, and 299.7 ± 63.2 ng/ml in patients with CKD and 594.6 ± 154.7 ng/ml, 354.5 ± 81.2 ng/ml, and 272.4 ± 91.8 ng/ml in healthy controls, respectively, (P < 0.001 vs. HD patients). MMP-2 was lower in patients with CKD: 405.6 ± 106.1 ng/ml compared with 516.9 ± 81.7 ng/ml in controls (P = 0.02). The MMP-2/TIMP-2 ratio was increased in HD patients compared with both patients with CKD and controls. In the course of an HD session, MMP-2 and TIMP-1 serum levels were significantly decreased from pre-HD 570.0 ± 256.5 and 178.8 ± 66.9 ng/ml to post-HD 492.6 ± 212.5 and 144.6 ± 44.2 ng/ml (P = 0.004 and 0.013, respectively). However, the MMP-9/TIMP-1 ratio increased from pre-HD 0.15 (2.19) (median, range) to 0.23 (0.33) after a HD session (P = 0.03). CRP was positively correlated with MMP-9 and MMP-9/TIMP-1 ratio in HD patients and patients with CKD (r = 0.67; P = 0.03). CONCLUSIONS: The MMP-9/TIMP-1 ratio increased during HD sessions, although their absolute levels were lowered. This change may represent a chronic state of enhanced fibrosis in patients undergoing HD.

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.

[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.

[Neutrophils and selected cytokines in ulcerative colitis pathomechanism]. / Udzial neutrofilów i wybranych cytokin w patomechanizmie wrzodziejacego zapalenia jelita grubego.

The aim of the study was to assess receptor CD11b and CD62L expression on neutrophils and IL-1beta, IL-6 and IL-8 level in children with ulcerative colitis. Thirty children aged 8-17 years were included in the study. The investigations were performed in the acute phase, prior to the treatment and during remission, 2 months after clinical improvement was obtained. The control group consisted of 12 children with chronic obstipation. IL-1beta, IL-6 and IL-8 serum level and receptor CD11b expression, especially on resting neutrophils, in children with severe and moderate course of the disease were statistically significantly higher; whereas CD62L expression was significantly lower in comparison with the controls and got back to normal during remission. Increased IL-1beta level was observed only in children with severe disease course. In children with mild process the results of the investigations were similar to the control group. In children with severe disease activity the proinflammatory cytokines levels in serum were elevated. The neutrophils in children with severe course of the disease manifested priming in peripheral blood before leaving the circulation.

Effects of cardiopulmonary bypass on the expression of adhesive molecules in children undergoing cardiac surgery due to congenital heart disorders.

BACKGROUND: Infants and small children who undergo cardiac surgery due to congenital heart disorders, are at risk of developing inflammatory complications and multi-organ failure. AIM: To assess the changes in the expression of adhesive molecules on neutrophils and in peripheral blood plasma in children undergoing cardiac surgery with or without the use of cardiopulmonary bypass (CPB). METHODS: In 18 children who underwent surgery with the use of CPB and in 7 children who underwent surgery without CPB, the expression of CD11b, CD11c and CD62L molecules on neutrophils and soluble adhesive molecules - L-selectin and ICAM-1, was assessed before, during and after cardiac surgery. RESULTS: A significant increase in the expression of CD11b on neutrophils at the time of the initiation of CPB was observed. The most pronounced decrease in the expression of CD11c was detected at the end of surgical procedure. There was also a negative correlation between L-selectin concentration measured at the end of CPB and CPB duration as well as between ICAM-1 concentration and duration of hypothermia. CONCLUSIONS: Cardiac surgery influences the expression of adhesive molecules on neutrophils and in blood plasma. These changes are more pronounced in children who undergo CPB, and depend on CPB duration as well as the use of hypothermia.

[Effect of cardiopulmonary bypass on selected neutrophil functions in children with cyanotic congenital heart disease]. / Wplyw krazenia pozaustrojowego na wybrane funkcje neutrofili u dzieci z wrodzona sinicza wada serca.

BACKGROUND: Cardiopulmonary bypass (CPB) is known to induce several pathogenic responses in cardiovascular surgery. Children suffering from cyanotic congenital heart disease (CCHD) have been assumed to have a deficiency in their defence mechanisms. The preoperative clinical condition (hypoxemia and circulation disorders) could initiate post CPB inflammatory response. The nature of the participation of neutrophils and its time course in the post CPB inflammatory response, however, is less clear. THE AIM of our study was to investigate alterations in expression of adhesion molecules and L-selectin on neutrophils, and soluble adhesion molecules levels in CCHD children. MATERIALS AND METHODS: 23 infants and children with congenital heart disease, scheduled for primary corrective surgical repair; were enrolled. Expression of CD11b. CD18, and CD62L on neutrophils was determined by cytometry. Additionally, serum concentrations of soluble ELAM-1, L-selectin were measured using enzyme-linked immunoassay. Blood samples of children with CCHD were collected at the following times: before anaesthesia, at the initiation of CPB, after 30 minutes of CPB, at the end of CPB, and 24 hours and 72 hours after surgery. RESULTS: The expression of CD11b molecule on neutrophils was increased significantly at the start of CPB and returned to the baseline after 24 hrs. In all patients, preoperative CD11b expression on neutrophils correlated with hypoxemia. Reduced CD62L expression on neutrophils and soluble L-selectin concentration were observed at the end of CPB. CONCLUSION: We conclude, that cardiac surgery with CPB has a suppressive effect on adhesion molecules expression in children with CCHD, These findings also suggest that the type of neutrophil response to cardiac surgery appears to depend on preoperative clinical condition.

[Cytotoxic activity of natural killer cells in children with chronic type C hepatitis]. / Aktywnosc cytotoksyczna komórek NK u dzieci chorych na przewlekle zapalenie watroby typu C.

UNLABELLED: The aim of this study was to evaluate the activity of NK cells in peripheral blood in children with chronic type C hepatitis, and to determine the correlation between the activity of NK cells and histopathological changes in the liver. The study included 25 children with chronic type C hepatitis. The control group consisted of 10 children without liver diseases in past medical history and normal activity of amino-transferases. In all children the activity of NK cells was evaluated in relation to leukaemic cells of erytroleukaemia K-562 without stimulation and after stimulation with IL-2 (in vitro), and the percentage and absolute count of NK cells in peripheral blood were determined by flow cytometry. RESULTS: In the studied group of children NK cells accounted for 11.68 +/- 6.73% of peripheral white blood cells, and their count was 241.08 +/- 128.56/ml. The cytotoxic activity of NK cells was 91.8 +/- 1.07% in solution E:T 25:1; and 6.72 +/- 3.68% in solution E:T 12.5:1. After stimulation with IL-2 it was 92.8 +/- 1.01% and 7.58 +/- 3.95%, respectively. The count of NK cells in serum and cytotoxic activity of NK cells in the studied group were not different from those of the control group. CONCLUSIONS: Key parameters of NK cells activity are not changed in children with chronic type C hepatitis.

[LDL-receptors expression on peripheral blood monocytes in various clinical types of atherosclerosis, depending on LDL concentration in serum]. / Ekspresja receptorów dla LDL na monocytach krwi obwodowej w róznych postaciach klinicznych miazdzycy.

In our studies an attempt was undertaken to establish whether there is a relationship between total cholesterol and LDL fraction concentrations and LDL receptors expression on monocytes in various clinical types of atherosclerosis and in subjects without clinical manifestations of this disease before and after 40 years of age. The study included 77 subjects divided into 4 groups: I--after myocardial infarction, II--after cerebral ischaemic stroke, III--with obliterative atheromatosis of the lower limbs, and control group IV without clinical symptoms of atheromatosis: a) subjects below 40 years of age b) over 40 years of age. Receptor expression on monocytes was tested with LDL Receptor Test (Orpegen Pharma, Heidelberg, Germany) using flow fluorometry method (FAC Scan, Backton Dickinson, San Jose, USA). The value of LDL receptors expression was calculated based on the difference of mean fluorescence intensity of monocytes incubated in low-lipid (LPDS) and autologous sera. It has been demonstrated that particular types of atheromatosis vary in the value of total cholesterol and LDL fraction concentrations. The highest concentrations of these lipid fractions were observes in ischaemic heart disease and only in this group they correlated with the value of LDL receptors expression on monocytes. This observation suggests that various clinical types of atheromathosis may differ pathogenically and lesions in the vessels may be the consequence of cholesterol transport defect or of lipid molecule modification; they not necessarily depend on the value of absolute lipid concentration in blood.

[Intrahepatic and peripheral blood NK cells in patients with chronic hepatitis C]. / Analiza wspólzaleznosci pomiedzy komórkami nk watroby i krwi obwodowej u dzieci z przewleklym zapaleniem watroby typu c.

UNLABELLED: The aim of this study was to evaluate the correlation between NK (natural killers) cells in an inflammatory infiltration of the liver and in peripheral blood in children with chronic hepatitis type C. MATERIAL AND METHODS: The study comprised 25 children with chronic hepatitis type C. The control group comprised 10 children with no liver disease in past medical history and normal activity of aminotransferases. In the study group, liver biopsy specimens were evaluated by histopathology and immunomorphology - the presence of NK cells present in the inflammatory infiltrate was determined. In all children the percentage and absolute count of NK cells in peripheral blood was evaluated by flow cytometry, as well as the activity of NK cells in relation to leukaemic cells of erythroleukemia K-562 without stimulation and after the stimulation with IL-2 (in vitro). RESULTS: In the studied group of children NK cells constituted 6.07 +/- 3.92 % of cells present in liver infiltrate. In peripheral blood NK cells constituted 11.68 +/- 6.73 % of white blood cells, and their absolute number was 241.08 +/- 128.56 /ml. The cytotoxic activity of NK cells in dilution E:T 25:1 was - 91.8 +/- 1.07 % and in dilution E:T 12.5:1 - 6.72 +/- 3.68 %. After stimulation with IL-2 it was 92.8 +/-1.01 % and 7.58 +/- 3.95 %, respectively. The number of NK cells in peripheral blood and cytotoxic activity of NK cells in the studied group did not differ from that of the control group. There was a positive correlation between the percentage of NK cells in the liver infiltrate and the absolute number of NK cells in the peripheral blood and between percentage of NK cells in the inflammatory infiltrate and cytotoxic activity of NK cells in peripheral blood, stimulated and non-stimulated with IL-2 in proportion E:T 12.5:1. CONCLUSIONS: Positive correlation between the number of NK cells in peripheral blood and liver infiltrate suggests the possibility of evaluating NK cells involvement in the pathogenesis of chronic hepatitis type C in children on the basis of peripheral blood tests.