The network of the Italian laboratories: a proficiency test on the quantification of trace elements in serum.

A proficiency test on the quantification of Al, Cu, Mn, Se and Zn in serum was carried out to verify the performance of about 30 regional laboratories of the network of Italian laboratories. The exercise consisted of four runs in which the laboratories were free in choosing analytical methods to determine trace elements in freeze-dried animal serum. Laboratories performances were evaluated by the study of statistical functions as Coefficients of Variation (CV), Youden plot and Z-score value. As for Al, the results were generally characterized by good accuracy and precision, in spite of the low levels of the element (5-7 microg l(-1)). Copper determination had some problems only at low concentration (about 160 microg l(-1)--first run), in which an elevated number of anomalous data were found. Better data were achieved for Zn, for which anomalous values were mainly stragglers than outliers. Due to the low number of data for Mn (concentrations from 0.6 to 60 microg l(-1)) and Se (concentration from 45 to 106 microg l(-1)), a restricted statistical treatment was applied; for these elements high CV values were found (range from 30 to 80%). The results of this trial confirmed that participation in a proficiency test represents a benefit for all analytical laboratories. In fact, with few exceptions, most of the participants improved their performances in terms of Z-score values.

A comparison of methods for the measurement of hemodialysis access recirculation.

BACKGROUND: NKF K-DOQI clinical practice guidelines recommend a regular monitoring of vascular access (VA) through several methods, such as VA recirculation (AR). Assessment of AR is crucial to avoid inefficiency of hemodialysis (HD). A non-urea based method has been developed (ultrasound dilution Transonics Hemodialysis Monitor--USM) which is considered the gold standard together with the two needle revised slow-stop-flow BUN recirculation method (S/SF). Furthermore, some other indicator dilution techniques, utilizing the dilution of serum potassium (K), glucose, hematocrit (Hct) and hemoglobin (Hb) have been recently described. METHODS: Aim of the study was to compare some of these tests with the gold standards (the USM and the revised S/SF methods). One hundred-five adult HD patients were selected to participate; all had autogenous radiocephalic wrist arteriovenous fistulas (AVF). Studies included the assessment of AR by means of: A. non-urea based methods: 1. at the start of HD a blood sample was obtained from the arterial needle at the time of needle insertion for the measurement of serum K, Hb and Hct (respectively K1, Hb1 and Hct1). The blood circuit was connected and the pumping of blood was started at 200 mL/min. After 18 seconds, blood samples were drawn from the arterial line sampling port (K2, Hb2, Hct2). At this time, if AR is present, part of the saline entering the blood stream will dilute K2, Hb2, Hct2.AR (%) is =100 x (1 - Hb2/Hb1) in the case of Hb; 2. during the first 30 min USM was performed in triplicate (the blood pump rate was set to 300 mL/min); B. a urea based method: the revised S/SF method was performed at 30 min. RESULTS: AR was absent when measured by means of the USM; it was absent even when cannulating the patients with the two needles placed as close as possible. AR measured by means of the USM was significantly different from AR measured by means of the other methods (means +/- SD, one-way analysis of variance, p < 0.0001). CONCLUSIONS: AR in autogenous radiocephalic wrist AVFs was zero when measured by means of the USM; the revised S/SF method and the tests employing the dilution of Hb, Hct and serum K can be considered a valuable alternative to the USM.

The impact of relapsing sterile icodextrin-associated peritonitis on peritoneal dialysis outcome.

BACKGROUND: The emphasis in peritoneal dialysis (PD) has shifted from a therapy with short-term goals to one of prolonging the life of the peritoneal membrane. Icodextrin (ICO), a starch-derived glucose polymer that is metabolized to maltose, is a valuable osmotic agent in the treatment of PD patients with defective ultrafiltration. However, ICO can cause sterile peritonitis. The manufacturer has recently withdrawn a series of batches of ICO solutions due to evidence of bacterial contamination (a bacterial cell wall breakdown, peptidoglycan). Some cases have been reported of culture-negative ICO-associated peritonitis which relapse on re-challenge. METHODS: We started to use ICO in chronic uremic PD patients in 1997. Ten patients out of 82 treated in our PD unit were exposed to ICO from 1997-2002. We registered 50 peritonitis episodes in this period: 34 were bacterial and 16 culture-negative. Among the 16 episodes of sterile peritonitis, 6 occurred in patients treated with ICO. Four of the 6 ICO treated patients experienced relapsing culture-negative episodes of peritonitis. We reviewed the records of the four patients. RESULTS: The first episodes of sterile ICO-associated peritonitis occurred between February-May 2002. These were clinically very mild and the only sign was abdominal discomfort and a cloudy dialysate containing a number of WBC/mm(3) ranging from 500-800. Cultures were negative. All the ICO solutions were from the batches withdrawn by the manufacturer. ICO (with new batches of solution) was re-introduced in all patients some weeks later to improve ultrafiltration. A new episode of sterile peritonitis occurred with the same characteristics as described above. Three of the four patients were re-challenged with new batches of ICO solution and again sterile peritonitis occurred. One patient was switched directly to hemodialysis (HD); the others were transferred to a program of automated PD including hypertonic glucose solutions. In 8 months all patients were switched to HD because of the failure of ultra-filtration. It is possible that the first episode of sterile peritonitis, associated with the use of an ICO batch suspected of having high peptidoglycan levels could have induced sensitization to ICO, which in turn could have been the cause of the relapse on re-challenge. CONCLUSIONS: The disappointing result of the relapsing culture-negative ICO-associated peritonitis in our patients was the unavoidable switch to HD, due to the inability of the hypertonic glucose solutions to ensure an adequate ultrafiltration. The moral of the story is that the pharmaceutical industry should market products that are more biocompatible and safer for chronic treatments such as PD.

Evaluation of five elements in lenses and aqueous humour of experimental rabbits after induced opacity.

We evaluated the presence of Ca, Na, K, Cu and Zn in the lenses and aqueous humour of rabbits treated with an Nd:YAG laser to induce opacity of the crystalline. The mean concentrations of the elements found in control lenses were: Ca: 15.8+/-5.2 mg/kg; Na: 1.2+/-0.6 g/kg; K: 10.3+/-3.3 g/kg; Cu: 0.19+/-0.06 mg/kg; Zn: 20.6+/-3.0 mg/kg. With the exception of K and Zn, the values found in the lenses of treated eyes (Ca: 135+/-24 mg/kg; Na: 4.3+/-1.5 g/kg; K: 10.1 +/- 3.2 g/kg; Cu: 0.47+/-0.17 mg/kg; Zn: 21.8+/-4.2 mg/kg) were significantly higher than in the controls. On the other hand, the concentrations found in the aqueous humour of treated eyes (Ca: 21.7+/-4.5 mg/l; Na: 0.66+/-0.21 g/l; K: 0.29+/-0.10 g/l; Cu: 0.035+/-0.009 mg/l; Zn: 0.079+/-0.01 mg/l) were significantly lower than those of the controls. The greatest difference was observed for Na (-68.6%) and Cu (-52.7%), followed by Ca and Zn (-35.0% and -35.2%, respectively). A positive correlation was found between Ca and Na in treated lenses (r2 = 0.9226, p < 0.0001) whereas inverse correlations were found for both Ca (r2 = 0.9788, p<0.0001) and Na (r2 = 0.9491, p<0.0001) between the concentrations found in the lenses and in the aqueous humour of treated eyes.

The natural history of autogenous radio-cephalic wrist arteriovenous fistulas of haemodialysis patients: a prospective observational study.

BACKGROUND: Clinical practice guidelines have supported vascular access surveillance programmes on the premise that the natural history of the vascular access will be altered by radiological or surgical interventions after vascular access dysfunction is detected. The primary objective of this study was to assess the actual risk of thrombosis of autogenous radio-cephalic (RC) wrist arteriovenous fistulas (AVFs) without any pre-emptive interventions. METHODS: We enrolled 52 randomly selected adult Caucasian prevalent haemodialysis (HD) patients, all with autogenous RC wrist AVFs, into this prospective, observational study aimed to follow the natural history of their AVFs for 4 years. The protocol prescribed avoiding any surgical or interventional radiological procedures until access failure (AVF thrombosis or a vascular access not assuring a single-pool Kt/V > or =1.2). The subjects underwent yearly assessments of vascular access blood flow rate by means of a saline ultrasound dilution method. RESULTS: All failures of vascular access were due to AVF thrombosis; none were attributed to an inadequacy of the dialysis dose. AVF thrombosis occurred in nine cases; a rate of 0.043 AVF thrombosis per patient-year at risk. A receiver operating characteristic curve, evaluating the diagnostic accuracy of baseline vascular access blood flow rate values in predicting AVF failure, showed an under-the-curve area of 0.82+/-0.05 SD (P = 0.01). The value of vascular access blood flow rate, identified as a predictor of AVF failure, was <700 ml/min with an 88.9% sensitivity and 68.6% specificity. When subdividing the population of AVFs into two groups according to the baseline vascular access blood flow rates, two out of the nine thromboses occurred among the AVFs that had baseline blood flow rates >700 ml/min (n = 31), whereas seven occurred among the AVFs that had baseline blood flow rates <700 ml/min (n = 21). The 4 year cumulative actuarial survival was 74.36 and 20.80%, respectively (log-rank test, P = 0.04). The 24 AVFs that remained patent at the end of the 4 years maintained a median blood flow rate > or =900 ml/min at all time points studied. Worth noting is that, five of them (20.8%) remained patent throughout the study with a blood flow rate consistently < or =500 ml/min. CONCLUSIONS: This study shows a very low rate of AVF thrombosis per patient-year at risk and a high actuarial survival of autogenous RC wrist AVFs, particularly of those having a blood flow rate >700 ml/min. Thus, a vascular access blood flow rate <700 ml/min appears to be a reliable cut-off point at which to start a closer monitoring of this parameter-which may lead to further investigations and possibly interventions relevant to the function of the AVFs.

Predictors of serum creatinine in haemodialysis patients: a cross-sectional analysis.

BACKGROUND: C-reactive protein (CRP) levels, an acute phase response index, predict cardiovascular outcome and are inversely related to visceral proteins, including albuminaemia in haemodialysis patients. Less definite is the relationship between inflammation and markers of somatic proteins such as serum creatinine in such patients. To explore these questions, a cross-sectional analysis of potential predictors of serum creatinine was performed. METHODS: One hundred and seventy-nine prevalent haemodialysis patients as of June 2001 were included in the cohort. Midweek pre-dialysis blood samples were collected during the months of June, September through to December 2001 inclusive, and determinations of serum urea (urease method), creatinine (alkaline picrate method) and CRP levels by means of a high sensitivity immunonephelometric method were performed. Furthermore, pre- and post-dialysis body weights were recorded and 2 min post-dialysis serum urea levels were determined three times. They were utilized for the calculation of single pool Kt/V and of normalized protein catabolic rate (nPCR). Each of the data represents the mean of three determinations made every 3 months in the study period. RESULTS: The analysis of multivariate linear regression was able to validate our model characterized by a dependent variable, serum creatinine and four independent variables (age, CRP, Kt/V and nPCR) (R(2)=0.60; F=24.10; P<0.00001; SE=1.94). Age (-0.08 mg/dl decrease in serum creatinine per 1-year increase in age), Kt/V (-0.25 mg/dl decrease in serum creatinine per 0.1 increase in Kt/V) and nPCR (0.10 mg/dl increase in serum creatinine per 0.1 g protein/kg/day increase in nPCR) were independently predictive of serum creatinine (P<0.00001). CRP and dialysis vintage did not predict serum creatinine. Stratifying the patients for the effects of CRP, only CRP values 4 mg/l were not. A further insight was given by the stratification of the patients for the effects of the interquartile ranges of CRP: it showed a progressive and statistically significant reduction of beta-coefficient inversely related to the increasing CRP values (P=0.003). Thus, the nature of the correlation between CRP and serum creatinine changes with increasing CRP values: from being a direct one, it shows a trend towards a transformation into an indirect one with beta=0 at a CRP value of approximately 9 mg/l. However, this indirect relationship does not reach statistical significance. CONCLUSIONS: The present cross-sectional study suggests that the activation of acute phase response does not influence creatinine metabolism in haemodialysis patients; in contrast, age, Kt/V and nPCR predict serum creatinine levels. Larger prospective trials are needed to achieve a definitive answer about the relationship between somatic proteins, acute phase response activation and nutrition in dialysis patients.

Renal osteodystrophy in predialysis and hemodialysis patients: comparison of histologic patterns and diagnostic predictivity of intact PTH.

BACKGROUND: Comparison of renal osteodystrophy in predialysis and hemodialysis has been rarely reported. Distinct patterns of renal osteodystrophy could be found in these conditions. In addition the use of parathyroid hormone (PTH) and other markers for noninvasive diagnosis may result in different predictive values in predialysis and hemodialysis patients. METHODS: 79 consecutive patients with conservative chronic renal failure and 107 patients on hemodialysis were studied. All patients were subjected to bone biopsy for histological and histomorphometric evaluation. The patients had no exposure to aluminium before dialysis and relatively low exposure while on hemodialysis. RESULTS: In the predialysis patients, bone biopsies showed 9 cases of adynamic bone disease (ABD) and 8 cases of osteomalacia (OM), 50 patients with mixed osteodystrophy and 2 cases of hyperparathyroidism. Among the hemodialysis patients 12 cases had ABD, 3 cases OM, 30 mixed osteodystrophy, and 61 patients hyperparathyroidism. In the predialysis patients with chronic renal failure, bone aluminium was on average 4.5 mg/kg dry weight, while in dialysis patients the average value was 35.4 mg/kg dry weight. Discriminant analysis of low turnover osteodystrophy (ABD and OM) by intact PTH showed higher accuracy in dialysis than in predialysis patients. Correlation studies of intact PTH versus bone formation rate, osteoblast surface/bone surface and osteoclast surface/bone surface showed significantly steeper slopes in dialysis than in predialysis patients, which indicates that bone resistance to PTH is more marked in predialysis patients. CONCLUSIONS: The prevalence of ABD and OM in the geographic area investigated is lower than in other reports. Aluminium exposure does not seem to be the cause of low turnover osteodystrophy in the present population. The predictive value of intact PTH in the noninvasive diagnosis of renal bone disease is higher in hemodialysis patients than in predialysis patients. Predialysis chronic renal failure, when compared to the dialysis stage, seems to be characterized by resistance of bone tissue to PTH.