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.

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.

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.

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.

[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.

Increased levels of soluble TNF-alpha receptors and cellular adhesion molecules in patients undergoing bioincompatible hemodialysis.

BACKGROUND: The study aimed to differentiate the effects of hemodialysis (HD) and chronic renal failure (CRF) on the levels of circulating tumor necrosis factor-alpha (TNF-alpha) and TNF-alpha receptors p55 and p75, soluble vascular cell adhesion molecule-1 (sVCAM-1), intercellular adhesion molecule-1 (sICAM-1), soluble endothelial-leukocyte adhesion molecule-1 (sE-selectin) and sP-selectin in 18 patients on regular HD treatment with cuprophane membrane in relation to 15 non-dialyzed CRF patients and 15 healthy controls. METHODS: The serum concentrations were determined with standard ELISA assays. RESULTS: Blood serum p75 and p55 were approximately tenfold increased in CRF (36.7 +/- 6.2 and 27.1 +/- 5.6 ng/ml) and HD patients (45.6 +/- 18.4 and 28.7 +/- 5.9 ng/ml) before the HD session (HD 0), during (HD 20) the session (45.7 +/- 18.4 and 28.5 +/- 7.3 ng/ml) and after (HD 240) the HD session (52.1 +/- 17.4 and 30.9 +/- 8.2 ng/ml) in comparison to control values (5.6 +/- 1.3 and 2.4 +/- 0.8 ng/ml, respectively) (p < 0.01). The highest increment of p75 at the end of HD session (HD 240) was also significantly higher than at preceding time points (HD 0 and 20) (p < 0.05). However, the remaining study parameters did not change during an HD session. Also, there were no relevant changes in TNF-alpha levels if (HD 0) 22.7 +/- 21.5 ng/ml and (HD 240) 21.1 +/- 18.9 ng/ml were compared. Chronic HD status was related to the increase of sVCAM-1 and sICAM-1 levels. Prior to HD, T0 sVCAM-1 and sICAM-1 concentrations were 2,180.4 +/- 761.8 and 567.3 +/- 218.8 ng/ml, during HD (T20): 2,172.7 +/- 759.2 and 602.3 +/- 379.9 ng/ml, and after HD (T240): 2,401.6 +/- 756.4 and 648.3 +/- 183.5 ng/ml, respectively (p < 0.05 vs. controls and CRF patients). sVCAM-1 and sICAM-1 serum levels (1,262.2 +/- 472.9 and 165.6 +/- 50.4 ng/ml) were similar in CRF patients and healthy controls (854.4 +/- 241.5 and 217.6 +/- 74.2 ng/ml, respectively). Even though serum sE- and sP-selectin in CRF patients did not differ from the control (39.8 +/- 21.3 vs. 42.1 +/- 18.9 ng/ml and 187.9 +/- 66.9 vs. 198.8 +/- 62.2 ng/ml, respectively), their levels were increased in HD patients up to 111.9 +/- 54.6 and 453.2 +/- 231.1 ng/ml in patients prior to HD, 118.7 +/- 66.2 and 350.8 +/- 114.8 ng/ml during the HD session and then 132.3 +/- 61.1 and 368.3 +/- 126.6 ng/ml, respectively, after its completion (p < 0.05 in comparison with CRF patients and controls). CONCLUSIONS: The increased circulating TNF-alpha receptors appear more associated with the uremic milieu than HD-related systemic inflammation, whereas increased soluble cellular adhesion molecules in patients undergoing bioincompatible HD may be related to the enhanced systemic inflammation specifically due to maintenance HD.

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.

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.

[Assessment of 8-hydroxy-2'-deoxyguanosine concentrations in bladder cancer patients treated with intravesical BCG instillation]. / Analiza stezenia 8-hydroksy-2'-deoxyguanozyny w moczu chorych na powierzchownego raka pecherza moczowego leczonych zawiesina pratka BCG.

UNLABELLED: Increased 8-hydroxy-2'-deoxyguanosine (8-OHdG) concentration has been proposed as reliable marker of the oxidative DNA damage, and prognostication of urological carcinogenesis, particularly in bladder cancer. Widely accepted method of treatment in early stages of bladder cancer is transurethral electro resection (TURN). Intravesical bacillus Calmette-Guerin instillation is considered to be the therapeutic agent for superficial transitional cell carcinoma (TCC) of the bladder and has been established as standard therapy in the patients. The aim of the study was to test the concentration of 8-OHdG in urine in patients with bladder cancer and the effect of TURN and BCG therapy on the 8-OHdG value. MATERIAL AND METHODS: We tested 12 patients (3 female and 9 male) with superficial bladder cancer and 31 healthy controls. Urine for the examination was drawn in 4 time points: before and 2 weeks after TURN treatment, then 6 weeks after 6 intravesical instillations of BCG (Onko BCG--BIO MED Lublin) by Morales and 12 weeks after TURN. 8-OHdG concentration in urine was tested using ELISA commercial kit (OXIS health) and the values of 8-OHdG are expressed as ng/ml of urine. RESULTS: Patients with superficial bladder cancer had 16.89 ng/ml of 8-OHdG in urine before the TURN procedure. The value was significantly (p< 0.005) higher then 12.98 ng/ml noted in healthy controls. 2 weeks after the procedure the 8-OHdG level decreased to 13.36 ng/ml. After 6 weeks of repeated 6 intravesical instillations of the BCG the concentration of 8-OHdG dropped to 10.91 ng/ml (p<0.005) and returned to the value 13.28 ng/ml after the next 4 weeks. CONCLUSIONS: Patients with bladder cancer have significantly increased concentration of 8-OHdG in urine compared to controls. 8 week combine therapy with TURN and BCG resulted in a significant decrease in 8-OHdG concentration in urine The beneficial effect of BCG instillations seems to result from strengthening of the antioxidative DNA protection.

Effect of cardiopulmonary bypass on neutrophil activity in pediatric open-heart surgery.

INTRODUCTION: The nature of the participation of neutrophils in the post-cardiopulmonary bypass (CPB) inflammatory response is not very clear. The aim of our study was to investigate alterations in neutrophil phagocytic activity and adhesion molecule expression on these cells in children during and after CPB. MATERIAL/METHODS: Twenty-one children aged 6-33 months with congenital heart disease, scheduled for pri mary corrective surgery, were enrolled. The expressions of CD11b adhesion molecules and Fc? receptor on neutrophils and their phagocytic activity were evaluated. The studied markers were sequentially measured before, at the initiation of, and after CPB. RESULTS: During the course of the operation, CD11b molecule expression on neutrophils showed a slight elevation at the start of CPB (876.5+/-104.8 mean fluorescence intensity, MFI, vs. 768.1+/-178.2; p = 0.0047), followed by a significant decrease to 689.01+/-166.7 MFI after completion of the procedure. The expression of CD11b molecule on neutrophils measured at the end of CPB inversely correlated with the duration of CPB (r = -0.68, p = 0.00059). The expression of CD16 antigen dropped significantly at the start of CPB (1164.6+/-307.3 MFI vs. 1327.4+/-345.3 MFI; p = 0.0007) and remained decreased until the end of CPB (814.0+/-198.1 MFI). CONCLUSIONS: These findings suggest that the characteristics of the neutrophil response to cardiac surgery appear to depend on many factors. We demonstrated a link between the duration of CPB and adhesion molecule expression on neutrophils.

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.

Effects of uraemia and haemodialysis on neutrophil apoptosis and expression of apoptosis-related proteins.

BACKGROUND: In haemodialysis (HD) patients, it is unclear whether increased apoptosis of neutrophils is due to uraemia or HD itself. The purpose of the current study was to assess the effect of uraemia and HD on the rate of apoptosis and apoptosis-related protein expression in whole blood neutrophils. METHODS: We employed a whole-blood micromethod to test spontaneous apoptosis and expression of apoptosis-regulating proteins in cultured neutrophils from uraemic patients (pre-HD), HD patients and healthy controls. Blood samples were drawn before, after 20 min and after 4 h of haemodialysis, and were then cultured for 20 h. We evaluated the rate of apoptosis from annexin V and propidium iodide staining, and examined bcl-2, Fas/Apo-1 and p53 expression in the cultured neutrophils. RESULTS: Fas/APO-1 expression and total percentage of apoptotic whole blood neutrophils of pre-HD and HD patients before HD were significantly higher than controls. There was a transient but significant decrease in the percentage of apoptotic neutrophils and Fas/APO-1 expression after 20 min of dialysis. The expression of bcl-2 protein was significantly lower from neutrophils in HD patients compared with controls, and HD significantly downregulated bcl-2 expression. The p53 protein content in HD patients before HD was significantly higher than in pre-HD patients. CONCLUSIONS: These findings suggest that uraemia accelerates neutrophil apoptosis by increasing Fas/Apo-1, and that HD does not affect neutrophil apoptosis more than uraemia. In addition, HD produces only in a transient sequestration of potentially apoptotic neutrophils.