Total, free, and protein-bound sulphur amino acids in uraemic patients

Fasting plasma concentrations of sulphur amino acids (sAA) were measured in none non-dialysed (ND) chronic uraemic patients on conservative treatment, 10 patients on continuous ambulatory peritoneal dialysis (CAPD), nine patients on haemodialysis (HD) treatment, and 10 healthy subjects (HS). Methionine and taurine concentrations were significantly decreased in the CAPD and HD patients and tended to be low in the ND patients. Cysteine sulphinic acid (CSA) levels were significantly higher in all patient groups. Total (t), free (f), and protein-bound (pb) homocysteine (Hcy) and cysteine (Cys) were significantly increased in all patient groups. Serine, a substrate for cystathionine synthesis from Hcy,, showed significantly lower concentrations in all patient groups. The percentages of pbHcy were significantly higher in the CAPD and HD patients than in the ND patients (P <0.0001, P=0.002 respectively) or in the HS (P <0.0001, P=0.008 respectively), whereas the percentages of pbCys in CAPD and HD patients were significantly higher than in ND patients (P=0.0006, P=0.009 respectively) and tended to be high without reaching statistical significance compared to the HS. A single HD treatment decreased tHcyby 26%, fHcy by 39%, and pbHcy by 22%, as well as tCys by 40%, fCys by 54%, and pbCys by 27%. The tHcy concentration, although decreased by HD treatment, remained higher than in HS, whereas tCys was normalized by the dialysis session. In addition, HD treatment significantly decreased the plasma concentrations of methionine, CSA, taurine, and serine. We conclude that, except for methionine and taurine, the plasma sAA in their different forms are markedly increased in dialysed and non-dialysed uraemic patients. The percentages of pbHcy and pbCys were significantly higher in dialysed than in ND uraemic patients. HD treatment can normalize the tCys concentration, and decrease the tHcy concentration but not normalize it. The observed hyperhomocysteineaemia and low taurine levels may contribute to the high incidence of cardiovascular disease in uraemic patients.     

Carotid artery intima-media thickness correlates with oxidative stress in chronic haemodialysis patients with accelerated atherosclerosis.

BACKGROUND: Accelerated atherosclerosis is the major cause of mortality in patients on chronic haemodialysis (HD). Increased oxidative stress might be the major factor leading to high cardiovascular mortality rate in HD patients. The aim of our study was to clarify effects of uraemia and dialysis on oxidative stress parameters and explore the relation between oxidative stress markers and carotid artery intima-media thickness (CIMT) as an indicator of atherosclerosis. METHODS: Twenty chronic HD patients, 20 predialytic uraemic patients and 20 healthy subjects were included in the study. Serum thiobarbituric acid reactive substances (TBARS), protein carbonyl content (PCO) and nitrite/nitrate levels were determined as oxidative stress markers. Serum vitamin E, plasma sulfhydryl (P-SH), erythrocyte glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities were measured as antioxidants. CIMT was assessed by carotid artery ultrasonography. RESULTS: Both chronic HD and predialytic uraemic patients had enhanced oxidative stress indicated by higher levels of nitrite/nitrate, TBARS and PCO, and lower levels of P-SH, SOD, CAT and GPx compared to controls. HD patients had significantly higher CIMT and nitrite/nitrate while significantly lower P-SH,vitamin E, SOD, CAT and GPx compared to predialytic uraemic patients. There was a significant positive correlation between CIMT and TBARS (r = 0.38, P = 0.003) and nitrite/nitrate levels (r = 0.41, P = 0.001), while there was a significant negative correlation between CIMT and SOD (r = -0.35, P = 0.01), CAT (r = -0.65, P < 0.001) and P-SH levels (r = -0.50, P < 0.001). A linear regression analysis showed that TBARS were still significantly and positively correlated with CIMT (P = 0.001), while CAT and P-SH were significantly and negatively correlated with CIMT (P = 0.002 and P = 0.048, respectively). CONCLUSIONS: HD exacerbates oxidative stress and disturbances in antioxidant enzymes in uraemic patients. We propose that serum TBARS and nitrite/nitrate can be used as positive determinants, while erythrocyte SOD, CAT and P-SH may be used as negative determinants of atherosclerosis assessed by CIMT in uraemic and HD patients.     

Triglyceride-rich lipoprotein abnormalities in CAPD-treated patients

In the present study intermediate-density lipoproteins (IDL)and very-low-density lipoproteins (VLDL) composition, structure,and mass were analysed in fasting uraemic patients on continuousambulatory peritoneal dialysis (CAPD) (n = 12) and on haemodialysis(HD) (n = 15), and in 15 healthy volunteers. All the groupswere matched for sex, age, and time on dialysis. Both groups of patients had elevated very-low-density lipoproteinlevels, CAPD patients four and HD group three times that ofcontrol. We found a fourfold and a twofold increase in the concentrationof IDL cholesterol in the CAPD and HD group respectively whenthey were compared with the control group. Both groups of patientspresent an increased VLDL mass. The CAPD group showed a four-foldincrease in IDL mass compared with the control group, whichindicated a preponderance of large size and suggested that defectiveIDL clearance was involved. The IDL composition of the CAPDpatients was significantly different from that of the HD patients:a twofold increase in IDL mass was observed in the CAPD patientsif compared with HD patients. We report new data concerningthe metabolism of triglyceride-rich lipoproteins in CAPD treatedpatients, which confirm the adverse effect of CAPD on serumlipids.     

Leptin in CAPD patients: serum concentrations and peritoneal loss.

BACKGROUND: To determine whether serum leptin concentrations in patients undergoing continuous ambulatory peritoneal dialysis (CAPD) are influenced by peritoneal loss of leptin and to compare serum leptin levels of normal subjects with those of patients receiving renal replacement therapy such as haemodialysis (HD), CAPD, or kidney transplantation. SUBJECTS AND METHODS: Eighty-four individuals were investigated: six females and 14 males on standard CAPD; 13 females and 13 males on chronic HD; 10 female and eight male kidney transplant recipients, and 10 female and 10 male subjects as controls. Morning serum, 8-h and 24-h samples of peritoneal fluid concentrated to 6-20-fold by Centricon 3 (cutoff 3000 daltons), and 24-h urinary concentrations of leptin were measured with commercial RIA (Linco Research, Inc., USA). Venous blood and peritoneal fluid samples of albumin, beta2-microglobulin, glucose, urea, and creatinine were determined by standard laboratory techniques. Serum insulin levels were measured by radioimmunoassay. RESULTS: Patients (men and women) on CAPD and after kidney transplantation exhibited significantly higher serum concentrations of leptin and leptin/BMI ratios than control subjects. These increased values did not reach statistical significance in HD patients. Serum leptin concentrations were correlated very significantly with BMI in all cases (r=0.380, P<0.001). Moreover, in CAPD patients (r=0.630, P<0.007) and in HD patients (r=0.668, P<0.005), but not in kidney transplant recipients or control subjects, significant correlations were observed between serum leptin and insulin concentrations. Residual renal function (RRF) in the range 0-12.8 ml/min and serum beta2-microglobulin levels in the range 7.9-47.1 mg/l did not influence serum leptin levels in CAPD and HD patients. As expected, leptin was detected in the peritoneal fluid of CAPD patients. Twenty-four-hour peritoneal loss (30.95+/-21.05 ng/min) and 24-h peritoneal clearance (0.01+/-0.01 ml/kg/min) of leptin account for only 3.9% of estimated whole-body leptin production rate and 0.7% of leptin clearance from plasma respectively. Twenty-four-hour urinary losses of leptin in CAPD patients were negligible, accounting for 5.6+/-1.8% (range 0.3-15.2%) of total (peritoneal and urinary) loss of this hormone. CONCLUSIONS: These findings suggest that serum leptin levels are not affected by continuous peritoneal loss of leptin during CAPD and that insulin resistance and hyperinsulinaemia contribute to elevated serum leptin concentrations in CAPD and HD patients. The aetiology of increased serum leptin levels in kidney transplant recipients is probably different from that in dialysis patients.     

Interleukin-8 in chronic renal failure and dialysis patients

A total of 105 patients participated in this study, including10 with chronic glomerulonephritis with normal renal function(CGN patients), 36 uraemic patients (CRF patients), 19 continuousambulatory peritoneal dialysis patients (CAPD) without peritonitis,three CAPD patients with peritonitis, 37 patients undergoingchronic haemodialysis (HD) divided into short-term HD, 15 patients;medium-term HD, 12 patients; and long-term HD, 10 patients.IL-8 and two other proinflammatory cytokines, IL-6 and TNFweretested using a specific immunoassay. IL-8, IL-6, and TNFc serumlevels were significantly increased in patients with chronicrenal failure compared to their levels in normal individuals(P<0.000l, P<0.05 and P<0.000l respectively). The mostpronounced incre ment in IL-8, IL-6 and TNF serum levels wasobserved in CAPD patients (P<0.000l). CAPD patients withoutperitonitis showed relatively low levels of IL-8 or IL-6 inperitoneal dialysate effluents (PDE), whereas PDE-TNF were notdetectable in almost all patients tested. Patients with peritonitisshowed very high serum and PDE levels of IL-8, IL-6 and TNF.The clinical recovery from peritonitis was characterized bya rapid fall in IL-8, IL-6 and TNF in serum and dialysate. HDpatients showed a significant increase in serum levels of IL-8and also IL-6 and TNFcompared to normal individuals (P<0.05,P<0.05 and P<0.01 respectively). HD duration influencedserum levels of IL-8 and TNF since they were significantly higherin short-term HD patients than medium- or long-term HD patients(respectively P<0.05, P<0.00l for IL-8, and P<0.01,P<0.001 for TNF Pre-HD IL-6 levels were not influenced byHD duration. No major modification of IL-8 serum levels couldbe evinced after and before HD sessions in the short-term group,but concentrations of this cytokine were significantly higherafter HD in medium- and long-term HD patients (P<0.05, P<0.0lrespectively). In contrast, HD session did not influence IL-6and TNF levels. We conclude that the cytokine profile is perturbedin uraemia and during dialysis, and that this should be consideredas an inflammatory status.     

Effects of uraemia and dialysis modality on polymorphonuclear cell apoptosis and function.

BACKGROUND: Previous studies have reported that incubation of polymorphonuclear cells (PMN) in uraemic plasma or with different haemodialysis membranes and peritoneal dialysis solutions increases apoptosis in this cell type. In addition, PMN harvested from uraemic patients show a reduced ability to generate superoxide in response to stimuli as well as impaired phagocytosis, chemotaxis and degranulation. The aim of the current study was to investigate the effect of uraemia and dialysis modality on apoptosis and function in freshly harvested non-incubated PMN. METHODS: Polymorphonuclear cells were harvested from 14 chronic haemodialysis (HD) patients, from 14 continuous peritoneal dialysis patients (CAPD), 28 chronic kidney disease (CKD), pre-dialysis patients and from 14 healthy subjects (Controls). In these in vivo experiments, PMN apoptosis was studied by means of flow cytometric analysis of annexin V binding to freshly isolated cells. Polymorphonuclear cell phagocytosis and production of reactive oxygen species by unstimulated or stimulated (S.aureus, fMLP, PMA) cells were also studied by flow cytometry using whole blood. RESULTS: We observed increased PMN apoptosis in CKD patients. CAPD and HD patients displayed PMN apoptosis rates similar to controls. In the HD group, PMN exhibited decreased phagocytosis rates. In contrast, phagocytosis rates in PMN from CAPD were not significantly different from controls. In the CKD and HD groups, apoptosis was inversely correlated with respiratory burst activity and phagocytosis. CONCLUSION: Our results suggest that both uraemia and treatment modality may interfere with PMN apoptosis and function. Dialysis appears to normalize the increased PMN apoptosis rates observed in CKD patients.     

Low-density lipoprotein subfraction profiles in chronic renal failure

Background: Small low-density lipoprotein (LDL) particle size, a newly recognized risk factor for cardiovascular disease in the general population, is frequently associated with hypertriglyceridaemia, the predominant plasma lipid abnormality present in uraemia. Methods: Plasma lipids and LDL subfraction profiles were examined in 33 non-dialysed patients with chronic renal failure (predial), 40 patients on continuous ambulatory peritoneal dialysis (CAPD), 42 haemodialysis patients (HD), 47 renal transplant recipients (RTR), and 44 controls. LDL subfractions separated by gel electrophoresis were scored by densitometric analysis (higher scores indicate profiles comprising smaller particles). Results: All groups with renal failure had significantly elevated (mean±SD) LDL scores (predial 1.36±0.6, CAPD 1.71±0.9, HD 1.68±0.9, RTR 1.92±0.8 vs control 0.87±0.4, all P<0.001), this being the only lipid abnormality detected in the predialysis patients. In CAPD and HD patients, LDL scores were associated with serum triglyceride (r=0.81, P<0.0001 and r=0.70, P<0.01 respectively), cholesterol (r=0.55, P<0.001 and r=0.49, P<0.01) and HDL-cholesterol (r=-0.43, P<0.01 and r=-0.51, P<0.01), whilst no such relationship was seen in the predialysis and RTR groups, suggesting that other factors were important. Conclusions: The presence of small LDL particles appears to be an early and unexplained feature of the uraemic dyslipidaemia. This abnormality persists after renal transplantation and may represent an important atherogenic risk factor. Key words: LD subfractions; renal failure; transplants      

Long-term CAPD patients are volume expanded and display more severe left ventricular hypertrophy than haemodialysis patients.

BACKGROUND: Whether hypertension and left ventricular hypertrophy (LVH) are more prevalent in CAPD than in haemodialysis (HD) patients is still under discussion. METHODS: To examine this problem we compared a group of 51 CAPD patients, with a group of 201 HD patients. The evaluation included the measurement of atrial natriuretic peptide (atrial natriuretic factor (ANF)), taken as indicator of volume status, and echocardiographic measurements. RESULTS: CAPD patients were older, had been treated for a shorter time, and had lower serum albumin and phosphate than HD patients. Plasma ANF was higher (P<0.01) in CAPD (median 33.8 pmol/l (interquartile range 18.2-63.0)) than in HD patients (22.7 pmol/l (14.9-38.7)). Similarly, the left atrial volume was substantially higher (P<0.0001) in CAPD patients (49+/-22 ml) than in HD patients (37+/-17 ml), while the left ventricular end-diastolic diameter was similar in the two groups (CAPD 51+/-7 mm; HD 50+/-7 mm). Furthermore, left ventricular hypertrophy was more severe (P<0.0001) in CAPD (157+/-37 g/m(2)) than in HD patients (133+/-39 g/m(2)). The proportion of CAPD patients requiring antihypertensive drugs was markedly higher than that of HD patients (65 vs 38% P<0.001). Multivariate modelling showed that volume expansion and pressure load as well as serum albumin were independent predictors of left ventricular mass. CONCLUSIONS: Left ventricular hypertrophy is more severe in long-term CAPD patients than in HD patients. This finding is associated with evidence of more pronounced volume expansion, hypertension, and hypoalbuminaemia. Volume and pressure load along with factors associated with hypoalbuminaemia may aggravate LVH in uraemic patients on CAPD.     

Plasma ghrelin levels in patients undergoing haemodialysis and peritoneal dialysis.

BACKGROUND: Ghrelin has been characterized as a relevant physiologic regulator of appetite and body weight in humans. However, the potential relationships between ghrelin levels, inflammation and malnutrition in dialysis patients have not been adequately studied. METHODS: We used a cross-sectional design to study 20 haemodialysis (HD) and 21 peritoneal dialysis (PD) patients, and compared their plasma ghrelin (PGhr) levels with that of an age-matched control group. We also explored correlations between ghrelin and selected hormonal, renal adequacy, nutritional and inflammation markers in both groups. RESULTS: PGhr levels were higher in HD (median 119.8 pg/ml, range 71.1-333.7, P = 0.001) and PD (99.3, range 45.8-578.5, P = 0.045) patients than in healthy controls (78, range 29-158) (HD vs PD, not significant). Ghrelin levels were strongly and inversely correlated with age (r = -0.46, P = 0.02 for patients; r = -0.61, P = 0.001 for controls). Except for a positive correlation between ghrelin and growth hormone (r = 0.48, P = 0.002), univariate analysis failed to detect associations between PGhr and the measured hormonal values, renal adequacy, nutritional indicators and markers of inflammation. However, multivariate analysis revealed significant inverse correlations between PGhr levels and nutritional markers, including subjective global assessment (P = 0.013), albumin (P = 0.001), transferrin (P = 0.01) and protein nitrogen appearance (as an estimate of protein intake) (P = 0.035), after controlling for the confounding effect of age. CONCLUSIONS: PGhr levels were moderately and similarly increased in patients undergoing HD and PD. Age was a strong determinant of PGhr levels, both in uraemic patients and in healthy controls. Dialysis adequacy, residual renal function and inflammation did not appear to influence ghrelin levels in these patients. The negative correlation between PGhr and nutritional markers suggests that low dietary intake causes increases in ghrelin secretion in dialysis patients.     

Mononuclear leukocyte apoptosis in haemodialysis patients: the role of cell thiols and vitamin E.

BACKGROUND: An increased apoptotic rate of peripheral blood mononuclear leukocytes (PBMLs) in haemodialysis (HD) patients has been reported in several studies, but its underlying mechanisms remain poorly understood. Oxidant stress is a well known cause of cell damage, and several lines of evidence suggest that it might influence the induction and signalling steps of mononuclear cell apoptosis through different mechanisms so as to provoke disturbances of the intracellular pool of thiols (SHi). In this study, we investigated the in vitro apoptotic rate and SHi of PBMLs in end-stage renal disease (ESRD) patients on HD or peritoneal dialysis (PD). METHODS: Apoptosis and SHi were evaluated in vitro in PBMLs obtained from 40 ESRD patients (HD, n = 30 and PD, n = 10) and 10 healthy controls. A subgroup of HD patients was also studied before and after 1 month of treatment with a vitamin E-coated dialyser (CL-E). Cell thiols and viability were also assessed in the monocyte-like cell line U937 and PBMLs after incubation in the presence of uraemic plasma with or without supplementation of the antioxidants vitamin E (70 micro M) or N-acetyl-cysteine (NAC) (0.5 mM). RESULTS: After 24 h in culture, the PBMLs of HD patients, but not those of CAPD patients, showed an apoptotic rate twice that of healthy controls and a 40% decrease of SHi levels (P < 0.01 in both). A negative correlation between the apoptotic rate and SHi was observed in both patients and controls (r = 0.648, P < 0.001). Plasma and ultrafiltrate samples from HD patients contained solutes (mainly in the low-middle molecular weight range) able to trigger apoptosis and oxidative stress in U937 cells. The treatment of HD patients with CL-E, as well as the in vitro supplementation of U937 cells with vitamin E or NAC during the exposure to uraemic plasma, decreased the rate of apoptosis and partially restored SHi. CONCLUSIONS: This study showed an association between an increased apoptotic rate and decreased SHi in PBML of HD patients, but not of CAPD patients. These changes are partially due to different pro-apoptogens that accumulate in the plasma and are at least partially prevented by exogenous antioxidants able to restore SHi, such as vitamin E or thiol suppliers.     

Haemolysis in haemodialysis patients: evidence for impaired defence mechanisms against oxidative stress.

BACKGROUND: Uraemic patients have a decreased ability to withstand oxidative stress. It is postulated that their antioxidant capacity is reduced, yet the mechanism remains unclear. Recently 33 haemodialysis (HD) patients were exposed to chloramine contamination in the water supply. This led to haemolysis in 24 patients, while nine were unaffected. In the former group haemoglobin decreased from 11.7+/-1.1 to 8.5+/- 1.4 g/dl (P<0.0001) and returned to 11.4+/-0.9 g/dl (P<0.0001) following recovery. During haemolysis, haptoglobin was 38.4+/-10.6 vs 138.1+/-8.3 ng/dl (P<0.0001) following recovery. METHODS: To explore the factors affecting the severity of haemolysis we studied extracellular and intracellular anti-oxidant defence mechanisms 3 months after recovery. In 29 patients and 20 controls we determined plasma glutathione (GSH), and the erythrocyte enzymes glutathione peroxidase (GSH-Px), glutathione reductase (GSH-Rx), and superoxide dismutase (SOD). Serum malondialdehyde (MDA) was measured as a marker of oxidative stress. RESULTS: Plasma GSH was lower in patients as compared to controls (5.49+/-0.26 vs 7.4+/-0.5 micromol/l, P<0.005). There was an inverse correlation between GSH and the degree of haemolysis (r=-0.42, P<0.02). Patients had higher GSH-Rx (4.64+/-0.15 vs 3.97+/-0.12 U/gHb, P<0.02), lower GSH-Px (29. 7+/-1.85 vs 35.5+/-1.62 U/gHb, P<0.001), and similar SOD (0.63+/-0. 02 vs 0.51+/-0.02 U/mgHb) as compared to controls. There was no correlation between the enzyme levels and the degree of haemolysis. MDA was higher in patients (2.37+/-0.07 vs 0.97+/-0.1 nmol/ml, P<0. 0001). There was a correlation between MDA and the years patients were on HD (r=0.43, P<0.02). CONCLUSIONS: These data indicate that HD patients have an impaired anti-oxidant response, which may be attributed in part, to plasma GSH deficiency. Patients with the lowest plasma GSH levels are more susceptible to oxidative stress and consequent haemolysis.     

Impact of dialysis therapy on insulin resistance in end-stage renal disease: comparison of haemodialysis and continuous ambulatory peritoneal dialysis.

BACKGROUND: Insulin resistance contributes to the pathogenesis of atherosclerotic cardiovascular disease and, thus, has an important impact on the mortality of uraemic patients. Haemodialysis (HD) is known to improve insulin resistance observed in uraemia. However, it is not known whether continuous ambulatory peritoneal dialysis (CAPD) alleviates insulin resistance in adult uraemic patients. The objective of this study was to compare the effect of two different dialysis modalities, HD and CAPD, on insulin resistance in adult uraemic patients and to identify the possible predictive factors for changes in insulin resistance. METHODS: Insulin resistance was examined in 19 non-diabetic patients with end-stage renal disease (ESRD) before and after dialysis therapy (HD, n=10; CAPD, n=9), as well as in 10 healthy controls using the hyperinsulinaemic euglycaemic glucose clamp technique. The glucose disposal rate (GDR mg/kg/min) was used as an index of insulin sensitivity during the clamp technique. We also determined which of various biochemical parameters might be associated with change in insulin resistance by carrying out multiple logistic regression analysis. RESULTS: GDR was significantly lower (6.44+/-1.76) in ESRD subjects than in normal subjects (9.90+/-2.01). HD and CAPD therapies significantly normalized GDR from 6.53+/-1.84 to 9.74+/-2.88 and from 6.35+/-1.65 to 8.18+/-1.76 respectively. Multiple logistic regression analysis showed that changes in BUN, haematocrit and plasma bicarbonate were significant predictive factors for the change in insulin resistance. CONCLUSION: CAPD therapy, in spite of its possible adverse effects in patients with atherosclerotic disease, has been shown to improve insulin resistance in adult uraemic patients, similarly to HD therapy.     

Diminished adhesion of CD4+ T cells from dialysis patients to extracellular matrix and its components fibronectin and laminin

Background: Cell-mediated immunity is impaired in uraemia. The recognition and ensuing interactions of immune cells, such as CD4+ T lymphocytes, with adhesive glycoproteins of the extra-cellular matrix (ECM) are mediated by integrins of the {beta}1 subfamily. We have previously demonstrated that uraemic sera inhibit the proliferation and adhesion of normal CD+ T cells to ECM components. In the present study, the adhesive capacity of CD4+ T lymphocytes of dialyzed patients (both haemodialysis [HD] and continuous ambulatory peritoneal dialysis [CAPD] was evaluated. Methods: Adhesion of CD4+ T cells from dialysis patients to intact ECM and its immobilized moieties, fibronectin (FN) and laminin (LN) was measured following phorbol-12-myristate-13-acetate (PMA) stimulation. In addition, cell surface expression of {beta}1 integrins (VLA 4-6) was determined by FACScan analysis. Results: Compared to normal cells, CD4+ T cells of dialysis patients demonstrated a significantly reduced adhesion to ECM, FN and LN (27-28 vs 52-55% P <0.001). This decreased adhesive capacity was not normalized upon incubation of the cells with normal sera. Cell surface expression of {beta}1 integrins was not modified. The inhibition of cell adhesion was more pronounced in CAPD patients (23-24% vs 29-30% in HD, P <0.02). Serum albumin correlated directly with cell adhesion. Aged HD patients' T cells demonstrated increased adhesion to ECM and its ligands, whereas a reverse trend was demonstrated in the CAPD group. Conclusions: T cells of dialysis patients exhibit abnormal adhesive activity, which may be due to an acquired cellular defect induced by uraemic milieu. CAPD patients show a greater degree of adhesion impairment, possibly due to their lower concentrations of serum albumin.     

Roles of paraoxonase and oxidative stress in adolescents with uraemic, essential or obesity-induced hypertension

BACKGROUND/AIMS: Paraoxonase 1 (PON1) is associated with high-density lipoproteins in the plasma, and is capable of hydrolysing oxidized lipids and preventing the oxidation of low-density lipoproteins. Oxidative stress and the PON1 (activity and Q192R polymorphism) were analysed in adolescent patients with essential (n = 49) or obesity-induced hypertension (n = 79), uraemic patients (n = 20), and also in obese normotensive patients (n = 60) and age-matched controls (n = 57). METHODS: The PON1 activity was measured via paraoxon hydrolysis. The PON1 genotype was determined by real-time PCR. The levels of oxidized and reduced glutathione, the end-products of nitric oxide, cysteine, homocysteine and lipid peroxidation in the plasma were measured and related to the PON1 status. RESULTS: There were no significant differences between the patient groups and the control group in the genotype distributions and the allele frequencies of the Q192R polymorphism. The PON activity was significantly lower (p < 0.001) in the uraemic hypertensive group than in the controls. The MDA concentration was significantly higher in the uraemic hypertensive (p < 0.001) and obese hypertensive (p < 0.05) patients. The plasma NOx concentrations were significantly lower (p < 0.001) and the ratio MDA/NOx were significantly higher in all four patient groups. The GSH levels were significantly lower in the patients with hypertension (p < 0.001) and obesity-induced hypertension (p < 0.05) than in the controls, while the GSSG level (p < 0.01) and the ratio GSSG/GSH (p < 0.05) was significantly higher in the uraemic hypertensive group. The plasma homocysteine level was significantly higher (p < 0.001) in the uraemic hypertensive patients as compared with the controls. CONCLUSIONS: We found no significant correlation between the biochemical parameters and neither genotypes nor enzyme activities. The PON1 status and the levels of certain biochemical parameters are independently associated with the hypertension in hypertensive and obese hypertensive patients, and the elevated levels of lipid peroxides and plasma homocysteine may contribute to the increased risk of cardiovascular complications in patients on haemodialysis.     

Erythrocyte defense mechanisms against free oxygen radicals in haemodialysed uraemic children

Changes in erythrocyte lipid peroxidation (measured as the concentration of malonyl dialdehyde), glutathione metabolism, antioxidant enzyme activities (glutathione peroxidase, catalase, and superoxide dismutase), the oxidized products of haemoglobin (Hb), and hydrogen peroxide (H₂O₂)-induced haemolysis were studied in six children with chronic renal failure treated with serial acetate and bicarbonate haemodialysis (HD). Ten age- and sex-matched children acted as controls. Malonyl dialdehyde levels were significantly higher and antioxidant enzyme activities lower in uraemic red blood cells (RBCs) compared with controls (P<0.05). Incubation of RBCs for 1 h with acetylphenylhydrazine induced a decrease in the concentration of reduced glutathione (P<0.001) and an increase in the level of oxidized products of Hb (P<0.001), but only in the uraemic patients. The H₂O₂ haemolysis test revealed a mild (n=3) to increased (n=3) haemolysis in the uraemic RBCs. Oxidative haemolysis is probably a multifactorial process in uraemic patients, and may be an important risk factor in HD therapy.     

Effect of recombinant human erythropoietin on synthesis of methylguanidine in uraemic patients on haemodialysis or continuous ambulatory peritoneal dialysis

The effect of recombinant human erythropoietin (rHuEPO) on synthesis of methylguanidine was studied in 6 uraemic patients on haemodialysis and 5 uraemic patients on continuous ambulatory peritoneal dialysis (CAPD). The Two groups of patients were started on a 24-week course of thrice weekly 1500 IU of rHuEPO by the intravenous route. Serum methylguanidine level and methylguanidine/creatinine ratio were comparable in these groups. In the two groups no significant differences were observed in these measurements comparing the pretreatment values with those 4, 8, 12 or 24 weeks after starting rHuEPO administration. During rHuEPO therapy, serum methylguanidine levels and methylguanidine/creatinine ratio showed no considerable difference between the two groups. These findings suggest that administration of rHuEPO does not alter methylguanidine synthesis in uraemic patients on haemodialysis and CAPD.     

A sensitive and specific ELISA for plasma pentosidine.

BACKGROUND: Advanced glycation end products are formed by non-enzymatic glycation and oxidation reaction. Pentosidine is a well-known and characterized structure among them, and has been implicated in the pathogenesis of complications associated with chronic renal failure and long-term dialysis, such as dialysis-related amyloidosis and atherosclerosis. METHODS: We established a highly sensitive and specific competitive enzyme-linked immunosorbent assay (ELISA) for plasma pentosidine and applied it to large numbers of plasma samples including haemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) patients. We compared their plasma pentosidine levels determined by the competitive ELISA with those determined by high-performance liquid chromatographic (HPLC) assay currently used. RESULTS: The plasma pentosidine levels determined by the ELISA were correlated well with those determined by sophisticated instrumental HPLC assay, both in non-diabetic and diabetic dialysis patients. Both analyses yielded comparable results, with over 8-fold higher plasma pentosidine levels in HD and CAPD patients, irrespective of the presence or absence of diabetes, as compared to normal subjects and non-uraemic diabetic patients. CONCLUSIONS: The competitive ELISA will provide a rapid and convenient determination of plasma pentosidine content and thus be useful to assess the carbonyl stress in uraemic patients.     

T-cell-independent and T-cell-dependent antibody responses in patients with chronic renal failure

Antibody responses against pneumococcal capsular antigens and tetanus toxoid were measured in 14 patients with chronic renal failure who were managed by continuous ambulatory peritoneal dialysis (CAPD) or haemodialysis (HD) and in eight healthy controls. IgG antipneumococcal responses were predominantly of the IgG2 and to a lesser extent IgG1 subclasses, while the IgG response against tetanus toxoid was largely IgG1 with smaller amounts of IgG4 and IgG3. The post-immunisation serum levels of IgG1 and IgM antibody against both antigens were significantly reduced in the uraemic patients compared with controls (P less than 0.05). All the uraemic patients had normal levels of IgG, IgA and IgM in the serum, but elevated levels of IgG3 prior to immunisation. The mechanisms responsible for the asymmetric depression of antibody responses in uraemia are unclear and may account in part for the increased susceptibility to infection in these patients.     

Parameters of oxidative stress and echocardiographic indexes in patients on dialysis therapy

Aim: Quantity of oxidative stress (OS) is enhanced in every stage of chronic renal failure (CRF). OS and its effects on echocardiographic indexes in patients on hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) were evaluated. Materials and methods: Thirty-nine patients on CAPD, 32 patients on HD, and 30 healthy individuals with similar demographic features were included. Patients with diabetes mellitus and chronic inflammatory diseases were excluded. Blood samples were collected to examine hematological and biochemical parameters and levels of malonyldialdehyde (MDA), glutathione peroxidase (GSH-px), and superoxide dismutase (SOD) after a 12-hour fasting period in the middle of dialysis week. OS parameters were compared with ejection fraction (EF), interventricular septum diameter (IVSd), left ventricular posterior wall diameter (LVPWd), and left atrium diameter (LAd) determined in M-mod echocardiographic examination. Results: No significant difference was observed between MDA and GSH-px levels of patients and control group; however, SOD levels of patients group were significantly lower (p < 0.0001). SOD levels of patients on HD were lower than that of patients on CAPD (p = 0.039). Negative correlation was detected between MDA and EF (r = ?0.380, p = 0.001); SOD has negative correlation with systolic blood pressure (r = ?0.265, p = 0.011), diastolic blood pressure (r = ?0.230, p = 0.028), phosphorus (r = ?0.327, p = 0.001), intact parathyroid hormone (iPTH) (r = ?0.259, p = 0.013), C-reactive protein (CRP) (r = ?0.235, p = 0.024), fibrinogen (r = ?0.342, p = 0.001), and total cholesterol (r = ?0.249, p = 0.017); and positive correlation with hemoglobin (r = 0.414, p < 0.001) and albumin (r = 0.367, p < 0.001). MDA was independently related with age (β = ?0.258, p = 0.035), male gender (β = ?0.312, p = 0.004), and EF (β = ?0.461, p < 0.001). No correlation was determined between antioxidants and cardiac indexes. Conclusion: SOD levels decreased significantly especially in patients on HD, and it was observed that lower levels of SOD would lead to OS in patients on HD and CAPD when compared to healthy individuals; MDA levels were independently influenced from EF.     

Increased renal allograft thrombosis in CAPD patients

In a retrospective analysis of 202 renal transplant proceduresin the years 1989–1992 we identified an excess of graftslost from primary renovascular thrombosis in patients receivingcontinuous ambulatory peritoneal dialysis (CAPD) compared tohaemodialysis (HD) patients (9 CAPD versus 0 HD, Chi-squared=9.63;P<0.01). All graft losses from thrombosis occurred within16 days of surgery. Possible predisposing causes were identifiedin three patients. Donor age was greater in CAPD patients losingtheir kidneys from thrombosis compared to the overall CAPD group[means (SD) years, 43.0(12.9) versus 29.1(15.8); P=0.01] whereasno significant difference in haematocrit, platelet count, antibodystatus, cyclosporin use, perioperative hypotension, primarydiagnosis, smoking, or diabetes mellitus was found. Data fromthe EDTA registry for 1990–91 show that graft loss fromprimary renovascular thrombosis in UK-treated patients was reportedin 7.1% of CAPD recipients compared with 1.8% in haemodialysis.We suggest that CAPD patients are at greater risk of graft lossfrom renovascular thrombosis than HD patients and may requiremore intensive fluid and anticoagulant treatment in the perioperativeperiod.