[Teaching Evidence Based Medicine EBM in medical faculties in Poland]. / Nauczanie Evidence Based Medicine (EBM) na wydzialach lekarskich w Polsce.

Evidence Based Medicine (EBM) is a core competence for clinicians and should be taught in all medical faculties. AIM: The aim of the study was to survey how EBM is taught in medical faculties in Poland. MATERIALS AND METHODS: We conducted a questionnaire study, asking the deans of medical faculties to fill it. RESULTS: We got a response from all medical faculties. Teaching of EBM is carried out in all medical faculties in Poland, apart from Kopernicus University in Torun. EBM is a separate subject in 7 faculties (from 5 to 36 hours). The most often EBM is taught on the IIIrd to the Vth course of the study with exception of Kielce, where it is held on the IInd course. In the most faculties teaching EBM is obligatory. In Lodz and Krakow, apart of being obligatory, EBM may be continued facultatively. Teachers are competent in EBM and continuosly trained. EBM study ends with a credit in 3 faculties. Some faculties intend to introduce EBM as a separate subject or extend number of hours. CONCLUSIONS: Our study showed that EBM is generally taught in medical faculties in Poland although in various length (5-36 hours). It should be extended to 30 hours and unified within a separate subject which ends with a credit.

The problem of Lyme borreliosis infections in urban and rural residents in Poland, based on National Health Fund data.

INTRODUCTION AND OBJECTIVE: Some fragmentary studies show that the incidence of Lyme borreliosis in Poland is increasing. It has been generally accepted that the most affected are forestry workers and farmers. The aim of the study is to compare the incidence of borreliosis in urban and rural residents in 2008-2016. MATERIAL AND METHODS: Databases on Lyme borreliosis from the National Health Fund and Central Statistical Office in Poland were analyzed. For each patient, ambulatory or discharged from every hospital, the diagnosis was compulsorily reported as encoded following the International Classification of Diseases. RESULTS: A steadily increasing number of patients with borreliosis in Poland was found, which doubled in 2008 - 2016. The incidence was similar in urban and rural residents. In all the provinces in Poland, an increase in incidence of borreliosis was observed, although there were big differences between them. The highest frequency of borreliosis was in Podlasie and Warmia-Masuria provinces. The lowest incidence of borreliosis was noticed in Wielkopolska province. In the most provinces the increase in the incidence of borreliosis was steady, except Warmia-Masuria, where it was very low in 2008, and soaring since 2011. The number of cases per year between 2008 - 2016 increased in both in males and females. CONCLUSIONS: The results suggest the need for higher awareness of the risk of Lyme borreliosis in urban residents, because the incidence of Lyme borreliosis is growing independently of the place of residence. Prompt measures to prevent tick bites and appropriate education are urgently needed.

Oxidative Storm Induced by Tryptophan Metabolites: Missing Link between Atherosclerosis and Chronic Kidney Disease.

Chronic kidney disease (CKD) occurrence is rising all over the world. Its presence is associated with an increased risk of premature death from cardiovascular disease (CVD). Several explanations of this link have been put forward. It is known that in renal failure, an array of metabolites cannot be excreted, and they accumulate in the organism. Among them, some are metabolites of tryptophan (TRP), such as indoxyl sulfate and kynurenine. Scientists have become interested in them in the context of inducing vascular damage in the course of chronic kidney impairment. Experimental evidence suggests the involvement of TRP metabolites in the progression of chronic kidney disease and atherosclerosis separately and point to oxidative stress generation as one of the main mechanisms that is responsible for worsening those states. Since it is known that blood levels of those metabolites increase significantly in renal failure and that they generate reactive oxygen species (ROS), which lead to endothelial injury, it is reasonable to suspect that products of TRP metabolism are the missing link in frequently occurring atherosclerosis in CKD patients. This review focuses on reports that shed a light on TRP metabolites as contributing factors to vascular damage in the progression of impaired kidney function.

Syphilis in Poland is on the rise and underreported.

INTRODUCTION: Despite the availability of diagnostic tests and effective treatment, there has been a problem with vigilance and reporting of that infectious disease in many countries including Poland. AIM: To compare the incidence of syphilis in Poland in years 2010-2016 according to the mandatory epidemiological surveillance system with the data of the National Health Fund (NHF). MATERIAL AND METHODS: Data of the NHF in Poland were collected. The total number of patients with syphilis (all forms) was estimated on the basis of their unique identifying numbers (PESEL). RESULTS: The steady increase in the incidence of syphilis in Poland throughout 2010-2016 was found, apart from the congenital form of the disease, which decreased since 2010. The higher prevalence of syphilis was noted in men. The number of hospitalized patients remained constant. According to the data of the NHF, the number of cases of syphilis in Poland was twofold higher as compared to the statistics of the mandatory epidemiological surveillance system (National Institute of Public Health - National Institute of Hygiene, NIPH-NIH), which was the basis of reports published up to date. CONCLUSIONS: Our work shows that there is a remarkable underreporting of syphilis in the mandatory epidemiological surveillance system in Poland, involving also hospitalized patients. The use of the data of NHF in the surveillance of syphilis in Poland is proposed.

The impact of antihypertensive pharmacotherapy on interplay between protein-bound uremic toxin (indoxyl sulfate) and markers of inflammation in patients with chronic kidney disease.

PURPOSE: Indoxyl sulfate (IS) is one of the most potent uremic toxins involved in chronic kidney disease (CKD) progression, induction of inflammation, oxidative stress, and cardiovascular diseases occurrence. It is proved that hypertension is a common CVD complication and a major death risk factor as well as contributes for decline in a renal function. The aim of our study was to investigate how implementing of antihypertensive therapy impact IS concentrations and the associations between IS and markers of renal function, inflammation and oxidative stress. METHODS: Study was conducted on 50 patients diagnosed with CKD and hypertension, divided into three groups: without hypotensive therapy (CKD-NONE), hypotensive monotherapy (CKD-MONO), and hypotensive polypharmacotherapy (CKD-POLI), and 18 healthy volunteers. The markers of inflammation [interleukin-6, tumor necrosis factor-alpha (TNF-α), high-sensitive C-reactive protein (hs-CRP), neopterin, ferritin], oxidative status [superoxide dismutase (Cu/Zn-SOD), antibodies against oxidized low-density lipoprotein (oxLDL-abs)], and selectins were determinate using immunoenzymatic methods. IS levels were assayed using high-performance liquid chromatography and other parameters were analysed using routine laboratory techniques. Then cross-sectional analysis was performed. RESULTS: Elevated levels of IS, indicators of kidney function, markers of inflammation and blood pressure values were observed in each CKD subgroups. There was no effect of antihypertensive therapy on IS levels between studied groups, as well as there was no clear relationship between IS and blood pressure values in each studied group. The positive associations between IS and Cu/Zn SOD, neopterin, hs-CRP, creatinine and neutrophils/lymphocytes ratio were observed in CKD-NONE and CKD-POLI subgroups. Additionally, in CKD-POLI group IS positively correlated with TNF-α, ferritin and neutrophils. In CKD-MONO group, IS was positively related to oxLDL-abs, neopterin, E-selectin and creatinine, whereas it was inversely associated with hs-CRP. CONCLUSIONS: Our study showed for the first time that the antihypertensive therapy has no impact on IS levels in CKD patients with hypertension. However, the introduction of the antihypertensive therapy modified the dependencies between IS and the studied markers of kidney function, inflammation, oxidative stress and hematological parameters that are crucial for mortality and morbidity amongst the CKD patients with hypertension.

Association between uremic toxin-anthranilic acid and fibrinolytic system activity in predialysis patients at different stages of chronic kidney disease.

PURPOSE: Chronic kidney disease (CKD) is an estimated risk factor for increased mortality and morbidity due to fibrinolytic system disturbances. Progressive loss of renal function leads to retention of uremic toxins. Anthranilic acid (AA) is a tryptophan-derived uremic toxin with multidirectional properties that can affect the hemostatic system. The goal of this study was to examine the association between AA and the parameters of fibrinolysis at different stages of CKD. METHODS: Patients with CKD were divided into two groups: mild-to-moderate (n = 20) and severe-to-end-stage CKD (n = 28). Seventeen healthy volunteers served as an additional control group. Parameters of fibrinolysis, inflammation, and monocytes activation were determined by ELISA immune-enzymatic kits. AA levels were evaluated using high-performance liquid chromatography. RESULTS: AA concentration and parameters of fibrinolysis: urokinase-type plasminogen activator (uPA), its soluble receptor (suPAR), tissue plasminogen activator (tPA), tissue plasminogen activator inhibitor-1 (PAI-1) and plasmin-antiplasmin complex (PAP) were significantly elevated in the CKD groups compared with the controls. The markers of inflammation, monocyte activation, and impaired kidney function were also increased in those with CKD. AA was positively correlated with the uPA/suPAR system in the early stages of CKD, whereas during severe-to-end-stage CKD, inverse relationships were observed between AA, tPA and PAI-1. Additionally, AA was an independent variable associated with tPA in patients with CKD overall and with uPA levels in the mild-to-moderate CKD group. CONCLUSIONS: Obtained results suggest for the first time the association between AA and the fibrinolytic system in CKD patients. The distinct relationship between AA and individual parameters of fibrinolysis appears to be dependent on CKD stage.

Unexpected and striking effect of heparin-free dialysis on cytokine release.

Heparin (both unfractionated and low molecular weight) is not only a potent anticoagulant but also has many pleiotropic effects, some of which are mediated by cytokine release. We compared the effect of hemodialysis (HD) with enoxaparin as an anticoagulant and without systemic anticoagulation (heparin-grafted membrane-Evodial) on the release of monocyte chemoattractant protein 1 (MCP-1), endostatin (ES) and activin A (Act-A). Nineteen stable HD patients were dialyzed with or without heparin, and plasma levels of MCP-1, ES and Act-A were measured after such a dialysis. During HD with Evodial, the intradialytic levels of all three cytokines were 2-3 folds lower. The between-anticoagulant differences were significant over time for all three cytokines: MCP-1 (P < 0.001), ES (P < 0.001) and Act-A (P < 0.001). This striking effect of heparin-free dialysis with Evodial membrane may be beneficial not only because it reduces the possibility of bleeding complications but also because it might reduce proinflammatory cytokine concentration and therefore contribute to the improvement in endothelial function. Further studies are needed to determine whether it has a positive effect on morbidity and mortality of maintenance HD patients.

Single center experience of subclinical rejections and BK nephropathies by kidney allografts' surveillance biopsies.

PURPOSE: Acute rejection of the kidney allograft remains the most important factor affecting the long-term graft outcome and is a major predictor of development of chronic damage and graft loss. Several studies have shown that early detection and treatment of subclinical rejection episodes may be beneficial for the graft outcome. The role of protocol (surveillance) biopsies and the value of donor specific antibodies (DSA) monitoring are still debatable. METHODS: This is a prospective observational study involving seventeen kidney recipients transplanted in north-eastern part of Poland who underwent "zero", 3-month and 12-month allograft biopsies as well as DSA assessment. RESULTS: Histologic analysis of the biopsies showed subclinical acute cellular rejection in 17.6% of patients (two tubulointerstitial, one vascular) at 3-months post transplantation, and additional case of borderline rejection at the 12-month point. Moreover, two cases (11.8%) of polyomavirus BK nephropathy were diagnosed (one at 3 and one at 12 month point). None of the patients developed de novo DSA. CONCLUSIONS: Our protocol biopsies allowed us to detect significant proportion of patients with subclinical, but histologically relevant acute cellular rejection and BK nephropathy. Early therapeutic intervention had beneficial effects in a 4-year follow up.

The effect of nephrectomy on Klotho, FGF-23 and bone metabolism.

BACKGROUND: Increased concentration of fibroblast growth factor 23 (FGF-23) and decreased levels of soluble Klotho (sKL) are linked to negative clinical outcomes among patients with chronic kidney disease and acute kidney injury. Therefore, it is reasonable to hypothesize that GFR reduction caused by nephrectomy might alter mineral metabolism and induces adverse consequences. Whether nephrectomy due to urological indications causes derangements in FGF-23 and sKL has not been studied. The aim of the study was to evaluate the effect of acute GFR decline due to unilateral nephrectomy on bone metabolism, FGF-23 and sKL levels. METHODS: This is a prospective, single-centre observational study of patients undergoing nephrectomy due to urological indications. Levels of C-terminal FGF-23 (c-FGF-23), sKL and bone turnover markers [ß-crosslaps (CTX), bone-specific alkaline phosphatase (bALP) and tartrate-resistant acid phosphatase 5b (TRAP 5b)] were measured before and after surgery (5 ± 2 days). RESULTS: Twenty-nine patients were studied (14 females, age 63.0 ± 11.6, eGFR 87.3 ± 19.2 ml/min/1.73 m2). After surgery, eGFR significantly declined (p < 0.0001). Nephrectomy significantly decreased sKL level [709.8 (599.9-831.2) vs. 583.0 (411.7-752.6) pg/ml, p < 0.001] and did not change c-FGF-23 concentration [70.5 (49.8-103.3) vs. 77.1 (60.5-109.1) RU/ml, p = 0.9]. Simultaneously, alterations in bone turnover markers were observed. Serum concentration of CTX increased [0.49 (0.4-0.64) vs. 0.59 (0.46-0.85) ng/ml, p = 0.001], while bALP and TRAP 5b decreased [23.6 (18.8-31.4) vs. 17.9 (15.0-22.0) U/l, p < 0.0001 and 3.3 (3.0-3.7) vs. 2.8 (2.3-3.2) U/l, p < 0.001, respectively]. CONCLUSIONS: Nephrectomy among patients with preserved renal function before surgery does not increase c-FGF-23 but reduces sKL. Moreover, nephrectomy results in derangements in bone turnover markers in short-term follow-up. These changes may participate in pathogenesis of bone disease after nephrectomy.

Indoxyl sulfate - the uremic toxin linking hemostatic system disturbances with the prevalence of cardiovascular disease in patients with chronic kidney disease.

BACKGROUND: During chronic kidney disease progression, kidney-specific risk factors for cardiovascular disease come into play. The present study investigated the impact of indoxyl sulfate, dietary tryptophan-derived uremic toxin, accumulated in the blood of patients with chronic kidney disease on hemostatic parameters, markers of inflammation, oxidative stress and monocyte to macrophage transition. METHODS: Fifty-one CKD patients not undergoing hemodialysis were enrolled in the study. Coagulation factors, fibrinolytic parameters, adhesion molecules, endothelial dysfunction markers, oxidative stress as well as inflammation markers were examined using immune-enzymatic method. Indoxyl sulfate levels were assessed using high-performance liquid chromatography. Biochemical parameters were determined by routine laboratory techniques using an automated analyzers. All assessed parameters were compared with controls and subjected to cross-sectional statistical analysis. RESULTS: Elevated concentrations of indoxyl sulfate, the vast majority of parameters affecting hemostasis, and markers of renal insufficiency conditions were observed. Part of hemostatic factors, namely tissue factor, von Willebrand factor, thrombomodulin, soluble urokinase-type plasminogen activator receptor, soluble intercellular adhesion molecule-1, soluble vascular cell adhesion protein were correlated with the fraction of indoxyl sulfate. A significant quantity of assessed parameters showed strong correlations with superoxide-dismutase, renal insufficiency rate, C-reactive protein, and neopterin. Levels of indoxyl sulfate were independently associated with markers of impaired endothelial function (thrombomodulin, adhesion molecules), oxidative stress (superoxide-dismutase) and monocytes activation determinant (neopterin), which indicate unconventional links between these systems and the role of indoxyl sulfate. Furthermore, parameters that correlated with the levels of indoxyl sulfate (von Willebrand factor, soluble urokinase-type plasminogen activator receptor, soluble intercellular adhesion molecule-1) were positively associated with the prevalence of cardiovascular disease in a CKD patients. CONCLUSIONS: The study demonstrated that in conditions of chronic exposure to uremic toxins, indoxyl sulfate seems to be one of the "missing links" between impaired renal function and prevalence of cardiovascular events, especially hemostatic disorders. The main functions of the action appear to be altered monocytes activation, intensified inflammatory process, and augmented oxidative stress by this uremic toxin.

Endocan--the new endothelial activation marker independently associated with soluble endothelial adhesion molecules in uraemic patients with cardiovascular disease.

OBJECTIVES: Endocan is a new marker of endothelial cell activation that mediates adhesion of leukocytes into endothelium. Soluble intercellular (sICAM-1) and vascular cellular (sVCAM-1) adhesion molecules play an important role in the prevalence of cardiovascular disease (CVD) in chronic kidney disease (CKD) patients. The aim of this study is to investigate whether endocan could affect the concentrations of sICAM-1 and sVCAM-1 in CKD patients, particularly in those with CVD. DESIGN AND METHODS: We evaluated plasma endocan, sICAM-1, sVCAM-1 and the markers of inflammation: high sensitivity C-reactive protein (hs CRP), interleukin-6, tumor necrosis factor-α (TNF-α) and their interrelationships in 53 CKD patients (both with and without CVD) and 29 healthy controls. RESULTS: Endocan, sICAM-1, sVCAM-1 and inflammatory markers were significantly higher in CKD patients than in controls, and patients with CVD had levels significantly higher (except interleukin-6 and TNF-α) than those without CVD. The presence of CVD, ferritin, TNF-α and SBP were the independent predictors of endocan levels in the whole CKD group. In this group, the weak relationship was between endocan and sICAM-1 and sVCAM-1, but age was the only independent predictor of these adhesion molecules. The strong association between endocan and sICAM-1 and sVCAM-1 was exclusively observed in subgroup with CVD, and the low % of lymphocytes followed by increased endocan was identified as the independent variables significantly associated with these soluble molecule levels. CONCLUSIONS: This study shows that plasma endocan is significantly increased and independently associated with sICAM-1 and sVCAM-1 levels in CKD patients with cardiovascular complications.

Cardiovascular risk in patients undergoing maintenance hemodialysis with Helixone® membrane: a multicenter randomized study.

INTRODUCTION: The high prevalence and incidence of atherosclerotic vascular complications, such as cardiovascular disease, remain the major cause of morbidity and mortality in patients undergoing dialysis. OBJECTIVES: The aim of the study was to evaluate cardiovascular risk factors in patients dialyzed with a high­flux polysulfone membrane (Helixone®) compared with those dialyzed with a low­flux polysulfone membrane. PATIENTS AND METHODS: This was a crossover randomized study including 90 hemodialysis patients. Group 1 was treated first with high­flux and then with low­flux membranes, while group 2, first with low­flux and then with high­flux membranes for 13 months. Clinical, biochemical, and echocardiographic data were evaluated at baseline and every 3 months during the study. RESULTS: After 6 months of high­flux dialysis, we observed a significant decrease in ß2­microglobulin, lipoprotein(a), C­reactive protein, and parathormone levels and an increase in serum albumin levels. Initially, both groups showed left ventricular hypertrophy. After 6 months of high­flux dialysis, we observed a tendency for an increase in the cardiac index and cardiac output and a decrease in isovolumic relaxation time. CONCLUSIONS: Our study showed that the use of high­flux dialysis with the Helixone® membrane, in comparison with low­flux dialysis with polysulfone membranes, improves middle-molecular clearance. In addition, we showed that a reduction in chronic inflammation during high­flux dialysis may decrease cardiovascular risk. However, further research with longer follow­up is needed to verify our echocardiographic findings.

Effect of diabetes and oxidative stress on plasma CCL23 levels in patients with severe chronic kidney disease.

INTRODUCTION: CCL23 is a new CC chemokine involved in leukocyte trafficking and inflammatory diseases. Inflammation, oxidative stress, and diabetes are common in patients with chronic kidney disease (CKD), particularly in those with cardiovascular disease (CVD). OBJECTIVES: The aim of this study was to evaluate CCL23 concentrations in patients with CKD and to identify the factors affecting its plasma level. PATIENTS AND METHODS: CCL23 levels, inflammatory markers (high-sensitivity C-reactive protein, interleukin 6, and tumor necrosis factor α) and oxidative stress markers (neopterin and Cu/Zn superoxide dismutase [Cu/Zn SOD]) were measured in the plasma of patients with mild-to-moderate CKD (group A) and severe CKD (group B) both with and without diabetes and in controls. RESULTS: CCL23 concentrations were higher in group B, particularly in patients with diabetes compared with controls (P <0.001) and group A (P <0.01). The inflammatory markers were increased in CKD patients but they were not correlated with CCL23 levels. In contrast, there were associations between CCL23 and oxidative stress markers and kidney function. The presence of diabetes, Cu/Zn SOD, and percentage of lymphocytes were found to be independent factors affecting CCL23 concentrations in the whole CKD group. In patients without diabetes, only Cu/Zn SOD was independently associated with CCL23. CONCLUSIONS: CCL23 levels were increased in patients with severe CKD and were strongly correlated with kidney function. The coexistence of diabetes and oxidative stress independently affected CCL23 levels, while the presence of CVD and inflammation had no impact on its concentrations.

oxLDL - the molecule linking hypercoagulability with the presence of cardiovascular disease in hemodialyzed uraemic patients.

INTRODUCTION: Patients with end-stage renal disease (ESRD) exhibit features of a hypercoagulable state, which may contribute to cardiovascular complications. Data from "in vitro" studies suggest that cell-free plasma lipids and lipoproteins may be capable to support thrombin generation. The aim of this study has been to establish the role of plasma oxidized LDL (oxLDL) in the coagulation activation in hemodialyzed (HD) patients with and without cardiovascular disease (CVD). MATERIALS AND METHODS: We examined relationship between a marker of coagulation activation - prothrombin fragments 1+2 (F1+2), and plasma oxLDL levels in 60 HD patients with and without CVD and in 20 healthy controls. RESULTS: F1+2 levels were significantly higher in HD patients than in controls, and they were higher in HD patients with CVD compared to those without CVD (p<0.001). Conversely, oxLDL levels were similar in HD patients with CVD and healthy controls, whereas this parameter was lower in HD patients without CVD when compared to controls and patients with CVD (both p<0.01). Close positive and independent association was between F1+2 and oxLDL levels in HD patients. Nitrates treatment and the presence of pyelonephritis were associated with reduced oxLDL as well as hypercoagulability in HD patients with cardiovascular complications. CONCLUSION: This study demonstrates the independent association between oxLDL and the marker of coagulation activation - F1+2 in HD patients. This new observation may offer a better understanding of the complex mechanism leading to hypercoagulability, which is markedly intensified in these patients, particularly in those with CVD.

Effects of enoxaparin on myeloperoxidase release during hemodialysis.

Myeloperoxidase (MPO) is a proteolytic and prooxidant enzyme largely assembled with the vascular wall, and a heparin-binding protein. We studied if low-molecular-weight heparin enoxaparin administered for hemodialysis (HD) anticoagulation causes systemic MPO activation. Plasma MPO levels were measured in patients undergoing maintenance HD with an intravenous bolus of enoxaparin. Patients were retested during HD employing dialyzers with heparin-grafted polyacrylonitrile membrane and no systemic enoxaparin administration. During enoxaparin-anticoagulated HD plasma MPO levels strikingly increased in all patients (8.6-fold at 10 minutes and 3.3-fold at 120 minutes, both P < 0.0001). The increments were directly associated with the enoxaparin dosage and strongly inversely with the predialysis levels of the enzyme. The increase in plasma MPO during systemic heparin-free HD was significantly less pronounced. Enoxaparin administered for HD anticoagulation induces a marked and dose-dependent increase in plasma MPO as a plausibly favorable result of the liberation of the enzyme from the vascular wall.

Use of short prosthesis segments for brachiocephalic arteriovenous fistulas in elderly hemodialysis population.

INTRODUCTION: The autogenous brachiocephalic fistula is a recognized secondary access for hemodialysis. However, veins in the antecubital fossa are often damaged, due to repeated venipunctures and subsequent scarring. Sometimes their anatomy does not enable successful arteriovenous fistula creation. In cases when the proximal part of the cephalic vein seemed patent, during ultrasound Doppler examination, we decided to use a short segment of 6 mm polytetrafluoroethylene graft to connect the vein with the brachial artery. We report our series of this procedure. MATERIAL AND METHODS: Over an 8-year period, 34 patients underwent such an operation. Grafts were anastomosed either to the end of the cephalic vein or to the side. The decision was made based on the vein condition: small-caliber veins were considered better for the end-to-side anastomosis. All procedures were performed under local anesthesia, and were well tolerated. RESULTS: Thirty-three fistulas were successfully cannulated at 2-8 weeks after the operation. Fistula patency rates were 84%, 73% and 55% at 12, 24 and 36 months. Comparison of two anastomosis types showed differences, 50% and 62.8% at 36 months, yet without statistical significance (p = 0.27, log-rank test). Fistula patency was not influenced by patient's age, sex or comorbidities. CONCLUSIONS: The described procedure provides satisfactory cumulative patency with an acceptable complication rate. It can enhance the number of cephalic veins used with its main advantages of simple surgical technique and low perioperative morbidity.

Effect of unfractionated and low-molecular-weight heparin on OPG, sRANKL, and von Willebrand factor concentrations during hemodialysis.

BACKGROUND: Endothelial dysfunction marker, von Willebrand factor (vWF), is physically connected with osteoprotegerin (OPG) in the Weibel-Palade bodies. We aimed to compare the effect of unfractionated (UFH) and low-molecular-weight (LMWH enoxaparin) heparin used as anticoagulants during hemodialysis (HD) on plasma levels and relationships of OPG, soluble receptor activator of nuclear factor κB Ligand (sRANKL), and vWF. METHODS: Totally 21 clinically stable chronic HD patients were randomly assigned to either enoxaparin (n = 10) or UFH (n = 11) anticoagulation and followed prospectively for 12 weeks before crossing over to the alternate therapy for further 12 weeks. The OPG, RANKL, and vWF levels were measured at T0, T10, and T180 of HD session after each period of evaluation. RESULTS: The baseline sRANKL level was higher under UFH treatment. Its over-HD level does not behave significantly different under enoxaparin and UFH treatment. Plasma OPG levels expressly changed during both enoxaparin (χ(2) analysis of variance [ANOVA] = 31.13, P < .016) and UFH (χ(2) ANOVA = 8.26, P = .016) anticoagulation, and its increment at T10 and T180 was significantly different between both the heparins. The main negative predictor of OPG concentration was the total cholesterol level (ß = -.51, P = .025). von Willebrand factor concentration remained stable during UFH anticoagulation, whereas constant, no significant increments were noticed, under enoxaparin treatment. After 10 minutes of HD, especially under enoxaparin use, a positive correlation between OPG and vWF increase was noticed (P = .03, R = .45). CONCLUSIONS: Impact of heparin on endothelial cells and simultaneously on OPG/RANK/RANKL axis reinforces the presumption of the pathophysiological linkage between bone mineralization and endothelial dysfunction in end-stage renal disease.

Validation of Mini Nutritional Assessment Scale in peritoneal dialysis patients.

INTRODUCTION: Malnutrition is a negative predictive factor for survival in end stage renal disease (ESRD) patients. Coincidence of malnutrition, inflammation and atherosclerosis (MIA syndrome) in the dialysis population is an exceptionally poor outcome event. Due to flexibility, ease of performance and reproducibility, clinical scales are of particular value in assessment of nutritional status in ESRD patients. The aim of the present study was to evaluate the clinical value of Mini Nutritional Assessment (MNA) in peritoneal dialysis (PD) patients. MATERIAL AND METHODS: Nutritional status was assessed in 41 peritoneal dialysis patients by means of the MNA scale and malnutrition inflammation score (MIS). Some other clinical and laboratory parameters associated with nutritional status were analyzed. Patients were followed up for 30 months. RESULTS: In the analyzed group of patients a good nutritional state was diagnosed in 22 patients (54%), risk of malnutrition in 17 (41%) and malnutrition in 2 patients (5%) based on the MNA scale. A strong correlation between MNA based nutritional status and MIS was found (r = -0.85, p < 0.01, ANOVA, p < 0.01). Differences in time on dialysis, body mass index, concentration of albumin, cholesterol and triglycerides were noted between at risk/malnourished and well-nourished (according to MNA) patients. Statistically significant factors determining survival of patients by Cox proportional hazard analysis were age (HR 1.07), being at risk/malnourished according to MNA (HR 5.7), MIS (HR 1.2), and albumin (HR 0.13). CONCLUSIONS: The MNA scale is a valuable, clinically suitable tool for assessment of nutritional status in peritoneal dialysis patients. Risk of malnutrition and malnutrition diagnosed by MNA identifies patients at high mortality risk.

Effect of unfractionated heparin, enoxaparin and sulodexide on the relations between secretion and expression of OPG, RANKL and vWF in HUVEC.

Heparin modulates function of vascular endothelium. We studied the effects of unfractionated heparin (UFH) vs. enoxaparin vs. sulodexide on the levels and gene expression of osteoprotegerin (OPG), Receptor Activator of Nuclear Factor kB Ligand (RANKL) and von Willebrand factor (vWF) in Human Umbilical Vein Endothelial Cells (HUVEC) culture. HUVEC were isolated from human umbilical vein by a standard method. The supernatant concentrations (ELISA) and gene expression (Real Time-PCR) of OPG, RANKL and vWF in HUVEC were determined after incubation with various concentrations of UFH, enoxaparin and sulodexide for up to 16 hours. In control HUVEC strong positive correlation between OPG and vWF levels was observed, whereas sRANKL negatively correlated with OPG and vWF levels. Only in control HUVEC a negative correlation between the supernatant level of vWF and its gene expression was found. Already the lowest concentration of UFH caused 2.5-fold increase in OPG gene expression while higher UFH concentrations substantially increased RANKL mRNA level. A negative correlation between the OPG and sRANKL concentration was noticed in supernatant HUVEC which were incubated with enoxaparine. In conclusion, the observed interrelationships between OPG, RANKL and vWF levels in unstimulated HUVEC support the presumption of the pathophysiological links between these proteins. Of the tested heparin formulas UFH seems to be the most potent in altering the OPG, RANKL and vWF axis.