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Acute intermittent porphyria is one of eight disorders arising from disturbances in heme biosynthesis where the precursors, 5-aminolevulinate and porphobilinogen, are elevated in plasma and urine. Attacks are characterized by severe abdominal pain, vomiting and/or obstipation, neurological manifestations, and psychological disturbances. The mainstay of treatment is hemin infusion to induce the negative feedback of heme synthesis. Hemodialysis is casuistically suggested as an alternative treatment. We present a case report of a 78-year-old male with acute intermittent porphyria and renal failure treated with peritoneal dialysis resulting in complete discontinuance of longstanding painful and disabling porphyria attacks., (© 2021 Wiley Periodicals LLC.)

Introduction: The aim of this study was to establish reference intervals for plasma cystatin C and creatinine in adults using the Gentians cystatin C method traceable to the international calibrator standard ERM-DA471/IFCC and a creatinine method traceable to the IDMS (Isotope Dilution Mass Spectrometry) creatinine reference method., Methods: Blood samples were collected from 304 healthy blood donors (152 men and 152 women between 17 and 66 years old) with 30-31 men and 30-31 women in each ten-year interval. Plasma cystatin C was analyzed using the Gentian Cystatin C assay on a Roche cobas c702 analyzer, and plasma creatinine was analyzed using the CREA Plus assay on the Roche Modular P analyzer., Results: The nonparametric reference intervals for plasma cystatin C were 0.58-1.00 mg/L in women (median 0.78 mg/L, range 0.56-1.06 mg/L) and 0.62-1.04 mg/L in men (median 0.79 mg/L, range 0.61-1.07 mg/L). The Mann-Whitney U test revealed no gender-related difference in plasma cystatin C (P = .21). A common reference interval in women and men was calculated to be 0.61-1.01 mg/L (median 0.79 mg/L, range 0.56-1.07 mg/L). The nonparametric reference interval for plasma creatinine was 52-89 μmol/L in women (median 69 μmol/L, range 52-92 μmol/L) and 61-108 μmol/L in men (median 86 μmol/L, range 56-118 μmol/L). The Mann-Whitney U test revealed a gender-related difference in plasma creatinine (P < .0001)., Conclusion: In conclusion, we have established reference intervals for plasma cystatin C and creatinine in adults using methods traceable to international standards., (© 2018 Wiley Periodicals, Inc.)

Background: Programs for population screening of colorectal cancer (CRC) have been implemented in several countries with fecal immunochemical testing (FIT) as the preferred platform. However, the major obstacle for a feces-based testing method is the limited compliance that reduces the clinical sensitivity for detection of participants with non-symptomatic CRC. Therefore, research approaches have been initiated to develop screening concepts based on biomarkers in blood. Preliminary results show that protein, genetic, epigenetic, and metabolomic components may be valuable in blood-based screening concepts, particularly when combinations of the various components appear to lead to significant improvements., Objectives: The protocol described in this paper focuses on the validation of concepts based on biomarkers in blood in a major population screened by FIT., Methods: In Part 1, participants will be identified and included through the Danish CRC Screening Program comprising initial FIT and subsequent colonoscopy to those with a positive result. Blood samples will be collected from 8000 FIT-positive participants, who are offered subsequent colonoscopy. Findings and interventions at colonoscopy together with personal data including co-morbidity will be recorded. Blood samples and data will also be collected from 6000 arbitrarily chosen participants with negative FIT. In Part 2, blood samples and data will be collected from 30,000 FIT-negative participants three times within 4 years. The blood samples will be analyzed using various in-house and commercially available manual and automated analysis platforms., Results: We anticipate Part 1 to terminate late August 2016 and Part 2 to terminate late September 2022. The results from Parts 1 and 2 will be presented within 12 to 18 months from termination., Conclusions: The purpose of this study is to improve the efficacy of identifying participants with neoplastic bowel lesions, to identify false negative participants, to identify participants at risk of interval neoplastic lesions, to improve the compliance in screening sessions, and to establish guidelines for out-patient follow-up of at-risk participants based on combinations of blood-based biomarkers., Competing Interests: Conflicts of Interest: None declared.

Backgroud: Hyaluronic acid (HA) is proposed as a marker of functional liver capacity. The aim of the present study was to compare a new turbidimetric assay for measuring HA with the current standard method., Methods: HA was measured by a particle-enhanced turbidimetric immunoassay (PETIA) and enzyme-linked immunosorbent assay (ELISA) in a 40-sample dilution series and 39 intensive care unit (ICU) patients. Agreement was assessed with Bland-Altman's method., Results: In the ICU patients, the median HA concentration was 159.0 ng/ml (interquartile range (IQR) 117.5-362.5 ng/ml) with ELISA and 157.5 ng/ml (IQR 92.5-359.6 ng/ml) with PETIA. The mean difference was 12.88 ng/ml (95% CI, -4.3 to 30.1 ng/ml, P = 0.14) and the 95% limits of agreement were -91.17 to 116.9 ng/ml. In the dilution series, the mean difference was -59.26 ng/ml (95% CI, -74.68 to 43.84 ng/ml, P < 0.0001) and the 95% limits of agreement were 35.23 to -153.8 ng/ml., Conclusion: We found random variation between the PETIA and ELISA test that could affect performance in a clinical context, but only to a lesser extent in a research context. The new clinical biochemistry assay for HA determination will allow for large studies of the clinical utility of HA., (© 2015 Wiley Periodicals, Inc.)

Nitrate may lower methane production in ruminants by competing with methanogenesis for available hydrogen in the rumen. This study evaluated the effect of 4 levels of dietary nitrate addition on enteric methane production, hydrogen emission, feed intake, rumen fermentation, nutrient digestibility, microbial protein synthesis, and blood methemoglobin. In a 4×4 Latin square design 4 lactating Danish Holstein dairy cows fitted with rumen, duodenal, and ileal cannulas were assigned to 4 calcium ammonium nitrate addition levels: control, low, medium, and high [0, 5.3, 13.6, and 21.1g of nitrate/kg of dry matter (DM), respectively]. Diets were made isonitrogenous by replacing urea. Cows were fed ad libitum and, after a 6-d period of gradual introduction of nitrate, adapted to the corn-silage-based total mixed ration (forage:concentrate ratio 50:50 on DM basis) for 16d before sampling. Digesta content from duodenum, ileum, and feces, and rumen liquid were collected, after which methane production and hydrogen emissions were measured in respiration chambers. Methane production [L/kg of dry matter intake (DMI)] linearly decreased with increasing nitrate concentrations compared with the control, corresponding to a reduction of 6, 13, and 23% for the low, medium, and high diets, respectively. Methane production was lowered with apparent efficiencies (measured methane reduction relative to potential methane reduction) of 82.3, 71.9, and 79.4% for the low, medium, and high diets, respectively. Addition of nitrate increased hydrogen emissions (L/kg of DMI) quadratically by a factor of 2.5, 3.4, and 3.0 (as L/kg of DMI) for the low, medium, and high diets, respectively, compared with the control. Blood methemoglobin levels and nitrate concentrations in milk and urine increased with increasing nitrate intake, but did not constitute a threat for animal health and human food safety. Microbial crude protein synthesis and efficiency were unaffected. Total volatile fatty acid concentration and molar proportions of acetate, butyrate, and propionate were unaffected, whereas molar proportions of formate increased. Milk yield, milk composition, DMI and digestibility of DM, organic matter, crude protein, and neutral detergent fiber in rumen, small intestine, hindgut, and total tract were unaffected by addition of nitrate. In conclusion, nitrate lowered methane production linearly with minor effects on rumen fermentation and no effects on nutrient digestibility., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Background: Intra-abdominal hypertension (IAH) often leads to abdominal compartment syndrome, which is followed by intestinal ischemia and associated with a high mortality. The diagnosis of abdominal compartment syndrome is difficult, and no valid biochemical markers are available. We conducted an experimental study on pigs to determine if D-lactate could be a useful biochemical marker of intestinal ischemia., Materials and Methods: A total of eight pigs (intervention group) underwent insufflation of carbon dioxide in the abdominal cavity to induce IAH and were compared with that of eight pigs (sham group) without IAH. Blood samples were taken from the portal and jugular veins at 0, 60, 120, 180, and 240 min after insufflation of carbon dioxide, and concentrations of D-lactate and L-lactate in the two groups were compared using an unpaired t-test., Results: The concentrations of D-lactate were increased in portal blood after 180 min of IAH (P = 0.036) and jugular blood after 240 min of IAH (P = 0.028) in the intervention group compared with those in the sham group. A similar tendency was found for L-lactate levels after 180 min of IAH (P = 0.032 and P = 0.017 for portal and jugular blood samples, respectively). Examination of the intestines revealed both macroscopic and microscopic signs of ischemia in all but one animal in the intervention group and only in one sham-pig., Conclusions: Our findings suggest that D-lactate could be a useful biochemical marker of intestinal ischemia induced by IAH., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Objective: The purpose of this study in humans was to examine the influence of the gastrointestinal tract and liver on the serum concentrations of cystatin C., Methods: Eighteen healthy volunteers and 28 patients suspected of having chronic intestinal ischemia underwent catheterization of the abdominal aorta and the central hepatic vein. Blood samples were taken simultaneously from the abdominal aorta and the central hepatic vein 60, 90 and 120 minutes after the start of the investigation. After the first blood sample, a standard liquid meal was ingested. Measurement of splanchnic blood flow was performed using the Fick principle with constant infusion of (99m)Tc-Bridatec. Angiography was performed at the end of the investigation., Results: The splanchnic blood flow increased significantly postprandially in the healthy volunteers and in the patients with normal angiography by 0.613-0.698 L/min and increased non- significantly in the patients with abnormal angiography (n = 5) by 0.135 L/min on average. ANOVA and the Bonferroni's multiple comparison test showed no significant difference between the means of cystatin C, creatinine or urea in the samples taken 60, 90 and 120 minutes after the start of the investigation in the abdominal aorta and the hepatic vein in the healthy volunteers or in the patients suspected of chronic intestinal ischemia with normal angiography., Conclusion: There was no indication of hepatic elimination of cystatin C, creatinine or urea. The serum concentrations of cystatin C, creatinine and urea in the central hepatic vein and the abdominal aorta were independent of the splanchnic blood flow.

Background: Rodent models have been used to evaluate aspects of liver regeneration. The aim of the present study was to investigate the natural history of liver regeneration in healthy rats., Methods: A 70% partial hepatectomy was performed in 64 rats. The animals were randomised into 8 groups and evaluated on postoperative days one to eight. Hepatocyte proliferation was evaluated by immunohistochemistry using unbiased stereological principles., Results: The mean rat body weight was 238 g (211-287). The mean weight of the resected liver was 6.3 g (5.2-7.3) and the estimated mean total liver weight was 8.9 g (7.4-10.4). Both liver weight analysis and regeneration rate showed an ascending curve, with a maximum slope on postoperative days 1-4, reaching a steady state on days 5-8. Hepatocyte proliferation (positive Ki-67 cell profiles pr. mm(2)) was high (250 cell profiles/mm(2)) on postoperative days 1-3 and tapered off on day 5., Conclusion: Seventy percent partial hepatectomy in healthy rats induces a rapid regenerative response and PODs 2, 4 and 8 seems optimal for assessing hepatic growth in future studies., (Copyright © 2013 Surgical Associates Ltd. Published by Elsevier Ltd. All rights reserved.)

Background: The aim of this study was to compare the ability of renal indicators [serum creatinine (SCr), cystatin C (SCysC)] and glomerular filtration rate (GFR)-models to discriminate normal and reduced renal function. As a single cut-off level will always lead to false classifications, we propose using two cut-off levels, dividing renal function into normal or reduced, with an intermediate "gray zone" of indeterminable results., Methods: Glomerular filtration rate was measured by plasma clearance of (51)Cr-EDTA (13.7-147.4 mL/min/1.73 m(2)) in 119 children (age range 2.3-14.9 years). Reduced renal function was defined as a GFR of  <82 mL/min/1.73 m(2). SCr, SCysC, age-normalized creatinine (SCr-ratio), and eight published GFR-models were compared for their ability to correctly classify renal function as normal or reduced. Cut-off levels were determined so as to give 99 % certainty outside the gray zone., Results: The multivariable GFR-models by Schwartz et al. (J Am Soc Nephrol 2009; 20:629-637) and Zappitelli et al. (Am J Kidney Dis 2006; 48:221-230) and two models by Andersen et al. [Am J Kidney Dis 2012; 59(1):50-57: body cell mass (BCM)-model and Weight-model] performed significantly better than all other variables (P < 0.01), with the BCM-model performing the best (P < 0.05). The SCr-based Schwartz formula and SCr-ratio both performed better than SCr and SCysC., Conclusions: Among the 119 children enrolled in this study and the renal indicators tested, the BCM-model had the best diagnostic performance in terms of screening for normal or reduced renal function, and the SCr-ratio was a superior diagnostic tool to both SCr and SCysC.

Study Design: Prospective cohort study., Objectives: To investigate the relationship between (51)chromium-ethylene-diamine-tetra-acetate ((51)Cr-EDTA) clearance, serum cystatin C (CysC), serum creatinine, creatinine clearance and estimated glomerular filtration rate (eGFR(MDRD), MDRD stands for modification of diet in renal disease) based on the serum creatinine in patients with complete or incomplete spinal cord injury (SCI) and to develop and evaluate a GFR-estimating equation using serum CysC., Settings: Spinal Cord Injury Unit, Viborg Regional Hospital, Viborg, Denmark., Methods: Ninety-eight men and 47 women with SCI were included in the study. Serum CysC levels were measured by an automated particle-enhanced nephelometric immunoassay, serum and urine creatinine levels were measured by an enzymatic method traceable to the IDMS creatinine reference method, and (51)Cr-EDTA clearance was measured by a multiple plasma sample method., Results: The area under the curves (AUCs) in the non-parametric receiver operating characteristics (ROC) plots for serum CysC were compared with serum creatinine and to eGFR(MDRD) and revealed a significant difference (P-value < 0.05) for all SCI patients. There was no significant difference between the AUC for serum CysC compared with the AUC for creatinine clearance. GFR (ml min(-1) per 1.73 m(2)) can be calculated from serum CysC values (mg l(-1)) using the equation eGFR(CysC) = 212·exp(0.914·CysC). The model accurately predicted the GFR of 88% of patients within ± 30% of the measured GFR, and it was able to predict the GFR of 50% of patients within ± 10% of the measured GFR., Conclusion: In patients with SCI, GFR can be estimated independent of age, sex and muscle mass by a newly developed equation based on a single serum CysC value.

Objective: The objective of this study was to evaluate whether remote ischemic preconditioning can protect kidney function in children undergoing operation for complex congenital heart disease., Methods: Children (n = 113) aged 0 to 15 years admitted for complex congenital heart disease were randomly allocated according to age to remote ischemic preconditioning and control groups. After exclusion of 8 patients, we conducted the analysis on 105 patients (remote ischemic preconditioning group, n = 54; control group, n = 51). Before surgery, remote ischemic preconditioning was performed as 4 cycles of 5 minutes of ischemia by inflating a cuff around a leg to 40 mm Hg above the systolic pressure. End points were development of acute kidney injury, initiation of dialysis, plasma creatinine, estimated glomerular filtration rate, plasma cystatin C, plasma and urinary neutrophil gelatinase-associated lipocalin, and urinary output. Secondary end points included postoperative blood pressure, inotropic score, and mortality, as well as morbidity reflected by reoperation and stays in the intensive care unit and hospital., Results: Overall, 57 of the children (54%) had acute kidney injury develop, with 27 (50%) in the remote ischemic preconditioning group and 30 (59%) in the control group (P > .2). Remote ischemic preconditioning was not associated with improvement in either any of the renal biomarkers or any of the secondary end points., Conclusions: We found no evidence that remote ischemic preconditioning provided protection of kidney function in children undergoing operation for complex congenital heart disease., (Copyright © 2012 The American Association for Thoracic Surgery. Published by Mosby, Inc. All rights reserved.)

Background: Aiming to develop a more accurate cystatin C-based model for estimation of glomerular filtration rate (GFR) in children, we hypothesized that inclusion of body cell mass (BCM) would increase the accuracy of the GFR estimate in comparison to a well-established GFR reference method., Study Design: Diagnostic test accuracy study., Settings & Participants: 119 children (mean age, 8.8; range, 2.3-14.9 years) referred for GFR measurement by chromium 51 ethylenediaminetetraacetic acid ((51)Cr-EDTA) clearance (mean GFR, 98; range, 13.7-147.4 mL/min/1.73 m(2))., Index Test: GFR estimations by the 2 prediction models resulting from theoretical considerations corroborated by forward stepwise variable selection: GFR (mL/min) = 0.542 × (BCM/SCysC)(0.40) × (height × BSA/SCr)(0.65) and GFR (mL/min) = 0.426 × (weight/SCysC)(0.39) × (height × BSA/SCr)(0.64), where SCysC is serum cystatin C level, BSA is body surface area, and SCr is serum creatinine level. The accuracy and precision of these models were compared with 7 previously published prediction models using random subsampling cross-validation. Local constants and coefficients were calculated for all models. Root mean square error, R(2), and percentage of predictions within ±10% and ±30% of the reference GFR were calculated for all models. Based on 1,000 runs of the cross-validation procedure, median values and 2.5th and 97.5th quantiles of the validation parameters were calculated., Reference Test: GFR measurement by (51)Cr-EDTA clearance., Results: The BCM model predicted 98% within ±30% of reference GFR and 66% within ±10%, which was higher than for any other model. The weight model predicted 97.5% within ±30% of reference GFR and 62% within ±10%. The BCM model had the highest R(2) and the smallest root mean square error., Limitations: Included only 9 children with GFR <60 mL/min/1.73 m(2). Lack of independent validation cohort., Conclusions: The novel BCM model predicts GFR with higher accuracy than previously published models. The weight model is almost as accurate as the BCM model and allows for GFR estimation without knowledge of BCM. However, endogenous methods are still not sufficiently accurate to replace exogenous markers when GFR must be determined with high accuracy., (Copyright © 2011 National Kidney Foundation, Inc. Published by Elsevier Inc. All rights reserved.)

Background: Intestinal ischemia is difficult to diagnose, and search for new biomarkers has led to interest in D-lactate, which arises from bacterial fermentation in the gastrointestinal tract., Methods: The superior mesenteric artery was clamped in eight pigs for 6 h to induce ischemia of the intestine. Eight sham-operated pigs served as controls. Systemic and portal plasma D- and L-lactate were sampled in 1 h intervals. L-LDH was inactivated prior to D-lactate measurement by addition of NaOH., Results: In systemic vein samples, we found a significant mean difference in the change of D-lactate from baseline to 6 h between the sham and intervention group (.007 ± .011 mmol/l vs. .030 ± .013 mmol/l, respectively) (P = .020). Both systemic and portal circulation levels of plasma L-lactate increased significantly between the two groups within an hour. The mean difference for L-lactate were -.020 ± .215 mmol/l and 1.440 ± 1.454 mmol/l in the sham and intervention group, respectively (P = .009)., Conclusion: L-lactate was found to be a marker of arterial-induced intestinal ischemia in both the systemic and portal circulation. There was no significant elevation of D-lactate at either site during the 6 h of ischemia., (Copyright © 2012 Surgical Associates Ltd. Published by Elsevier Ltd. All rights reserved.)

Introduction: Our objective was to compare the bone and renal effects among HIV-infected patients randomized to abacavir or tenofovir-based combination anti-retroviral therapy., Methods: In an open-label randomized trial, HIV-infected patients were randomized to switch from zidovudine/lamivudine (AZT/3TC) to abacavir/lamivudine (ABC/3TC) or tenofovir/emtricitabine (TDF/FTC). We measured bone mass density (BMD) and bone turnover biomarkers (osteocalcin, osteocalcin, procollagen type 1 N-terminal propeptide (P1NP), alkaline phosphatase, type I collagen cross-linked C-telopeptide (CTx), and osteoprotegerin). We assessed renal function by estimated creatinine clearance, plasma cystatin C, and urinary levels of creatinine, albumin, cystatin C, and neutrophil gelatinase-associated lipocalin (NGAL). The changes from baseline in BMD and renal and bone biomarkers were compared across study arms., Results: Of 40 included patients, 35 completed 48 weeks of randomized therapy and follow up. BMD was measured in 33, 26, and 27 patients at baseline, week 24, and week 48, respectively. In TDF/FTC-treated patients we observed significant reductions from baseline in hip and lumbar spine BMD at week 24 (-1.8% and -2.5%) and week 48 (-2.1% and -2.1%), whereas BMD was stable in patients in the ABC/3TC arm. The changes from baseline in BMD were significantly different between study arms. All bone turnover biomarkers except osteoprotegerin increased in the TDF/FTC arm compared with the ABC/3TC arm, but early changes did not predict subsequent loss of BMD. Renal function parameters were similar between study arms although a small increase in NGAL was detected among TDF-treated patients., Conclusion: Switching to TDF/FTC-based therapy led to decreases in BMD and increases in bone turnover markers compared with ABC/3TC-based treatment. No major difference in renal function was observed., Trial Registration: Clinicaltrials.gov NCT00647244.

Background: Intestinal ischemia is difficult to diagnose. The search for biomarkers has led to an increased interest in d-lactate. d-lactate measured in higher concentrations arises from bacterial fermentation in the gastrointestinal tract. Permeable intestinal wall is an early consequence of intestinal ischemia, which allows d-lactate to enter the portal circulation., Methods: The superior mesenteric vein was clamped in eight pigs for two hours to induce ischemia of the intestine. Eight sham-operated pigs served as controls. Systemic and portal plasma d- and l-lactate, l-LDH and leukocytes were measured., Results: Plasma d-lactate increased significantly and nearly simultaneously in the systemic and portal circulation. After 75 min, samples from the jugular vein showed concentrations of .019 ± .008 mmol/L in the sham group and .042 ± .022 mmol/L in the intervention group (P = .023). A similar significant effect was seen in the portal circulation after 90 min. l-lactate increased five minutes after clamping in the portal circulation, with values of 3.396 ± 1.119 mmol/L in the intervention group compared to 1.696 ± .483 mmol/L in the control group (P = .006). l-LDH increased significantly in the intervention group, while leukocytes were unaffected. l-LDH and l-lactate in plasma led to an overestimation of d-lactate if not handled., Conclusion: Both systemic d- and l-lactate were markers of venous-induced intestinal ischemia. We speculate that d-lactate may be a valuable aid to the clinician in search of the anaerobic focus, because it may be more specific for mesenteric ischemia than l-lactate, in the sense that it is of bacterial origin., (Copyright © 2011 Surgical Associates Ltd. Published by Elsevier Ltd. All rights reserved.)

Objective: To evaluate the performance of the Roche Diagnostics Tina-quant Cystatin C particle enhanced immuno turbidimetric assay for the measurement of plasma and serum cystatin C, and to establish reference intervals for cystatin C in healthy blood donors., Methods and Materials: The cystatin C measurements were performed on the Roche Modular Analytics P automated clinical chemistry analyzer., Results: The cystatin C assay was linear in the measuring range 0.40-7.00 mg/L. Within-run CVs < or = 2.0%, between-run CVs < or = 4.2%, and total CVs < or = 5.5% in plasma pools and in commercial cystatin C control materials (range 1.0-4.7 mg/L). Recovery was 99.4-109.3%. No interference was detected from haemoglobin < 0.9 mmol/L, bilirubin < 330 micromol/L and Intralipid < 20 g/L. Measurement of cystatin C in Li-heparin plasma did not differ significantly from cystatin C measured in serum. Forty patient samples run on the Modular Analytics P (y) were compared to the Siemens Cystatin C assay on the BN II (x): y = 0.817x + 0.270, Sy.x = 0.168 (Deming regression). The non-parametric reference interval for cystatin C was calculated to be 0.41-0.91 mg/L in females (n = 86), and 0.43-0.94 mg/L in males (n = 76). The Mann-Whitney U test showed a significant difference between the two genders (p = 0.015), but the difference was without clinical relevance. A common reference interval for both genders (n = 162) was calculated to be 0.41-0.92 mg/L., Conclusion: The performance of the Tina-quant Cystatin C assay was acceptable for clinical use.

Objective: To evaluate the day-to-day biological variation of cystatin C in comparison with creatinine in healthy subjects and in patients with impaired renal function., Material and Methods: Eight weekly morning blood samples were taken from 20 healthy subjects (13 females and 7 males, median age 44 years, range 25-61) and 19 patients with impaired renal function (8 females and 11 males, median age 61 years, range 35-70). Serum cystatin C was measured using Dade Behring N Latex Cystatin C assay and serum creatinine by an enzymatic method (Roche)., Results: In the healthy subjects mean serum cystatin C was 0.70 mg/L (range 0.44-1.09) and mean serum creatinine 77 micromol/L (range 54-100). The analytical variance was 2.0% for cystatin C and 1.6% for creatinine. The intra-individual variance was greater for cystatin C than for creatinine (8.6% vs. 4.7%). The inter-individual variance was similar for both analytes (cystatin C 15.1% vs. creatinine 14.4%). In the patients with impaired renal function mean serum cystatin C was 1.86 mg/L (range 0.45-3.31) and mean serum creatinine 224 micromol/L (range 103-430). The analytical variance was 1.8% for cystatin C and 1.4% for creatinine. The intra-individual variance was greater for cystatin C than for creatinine (16.0% vs. 8.9%)., Conclusion: In the present study, the intra-individual variance was greater for cystatin C than for creatinine in both healthy subjects and in patients with impaired renal function. Accordingly, serum creatinine is the preferred marker for serial monitoring of renal function in individuals with stable muscle mass.

Cystatin C is a low molecular weight protein and the plasma level of cystatin C is mainly determined by glomerular filtration, making cystatin C an endogenous marker of glomerular filtration rate. The aim of the study was to elucidate the applicability of plasma cystatin C as a marker of renal function in patients with liver cirrhosis. Serum cystatin C and creatinine concentrations were compared with creatinine clearance. Thirty-six patients (14 females and 22 males aged between 33 and 81 years) with liver cirrhosis with normal to severely impaired kidney function were included. Plasma cystatin C was measured by an automated particle-enhanced nephelometric immunoassay (Dade Behring Diagnostics) and plasma creatinine by an enzymatic method. Plasma levels of cystatin C and creatinine were found to increase with decreasing values of creatinine clearance. The reciprocal values of cystatin C and creatinine were compared with those for creatinine clearance revealing an r2 of 0.37 and 0.18, respectively. Comparison of the areas under the curves (AUC) of the non-parametric receiver-operating characteristic plots for plasma cystatin C (AUC=0.7364; SE=0.0929) and plasma creatinine (AUC=0.6309: SF=0.1028) revealed a significant difference between plasma cystatin C and plasma levels of creatinine (p-value=0.03). The results demonstrate that the diagnostic accuracy of plasma cystatin C was better than plasma creatinine in identifying liver cirrhotic patients with reduced glomerular filtration rate.

Background: The cysteine protease inhibitor cystatin C may play a role in the development and progression of abdominal aortic aneurysms (AAAs)., Methods: From a mass screening trial of men aged 65-73 years, 151 small AAAs were followed for a mean of 2.9 years. Of these patients, 142 had serum samples taken to determine the levels of cystatin C, creatinine and C-reactive protein (CRP)., Results: Serum cystatin C concentration correlated negatively with AAA size (r = - 0.22 (95 per cent confidence interval (c.i.) - 0.59 to - 0.02)) and annual expansion rate (r = - 0.24 (95 per cent c.i. - 0.75 to - 0.05)), persisting after adjustment for renal function, smoking, diastolic blood pressure, CRP, age and AAA size. Creatinine clearance and CRP did not correlate with size or expansion rate. Thirty-one AAAs had expanded to over 50 mm, when operation was recommended. The serum level of cystatin C was a significant predictor of this occurrence, with a sensitivity and specificity of 61 and 57 per cent respectively. However, initial AAA size had the optimal sensitivity and specificity (both 81 per cent) in this regard., Conclusion: Deficiency of cystatin C was associated with increased aneurysm size and expansion rate, possibly due to lack of inhibition of cysteine proteases.

Unlabelled: Hepatocyte growth factor (HGF) is a growth-promoting peptide that appears to act in a renotropic and nephroprotective manner during acute renal damage. Recent studies suggest that HGF is also of importance in chronic renal diseases. The serum HGF level is correlated with serum creatinine, and it has been suggested that glomerular and tubular diseases affect serum HGF differently. In the present study. levels of serum HGF were determined and correlated to glomerular filtration rate (GFR) in 118 patients with various chronic renal diseases. GFR was determined by 99mTc-DTPA clearance, and the GFR values were evenly distributed in the interval 5-155 mL/min/1.73 m2. Serum HGF levels increased slightly with decreasing GFR: the Pearson correlation coefficient being 0.49 (p<0.0001). In 21 additional patients with end-stage renal disease treated with continuous ambulatory peritoneal dialysis, there was a more marked increase in the serum levels of HGF. The effect of glomerular and tubular diseases on serum HGF was examined by comparing the HGF levels in two groups of patients with similar GFR values: 57 patients with mainly glomerular disorders (diabetic nephropathy with micro- or macroalbuminuria or glomerulonephritis) and 14 patients with mainly tubular disorders (polycystic kidney disease). There was no significant difference between the HGF levels of the two groups (p=0.30)., In Conclusion: Serum HGF levels are correlated with GFR (for GFR > or = 5 mL/min/1.73 m2) in patients with chronic renal diseases, and glomerular and tubular disorders seem to affect the HGF level similarly.

Determination of porphobilinogen deaminase (PBGD; EC 4.3.1.8) activity in erythrocytes can contribute to the identification of patients suspected of acute intermittent porphyria. PBGD catalyses the polymerization of four molecules of porphobilinogen (PBG) to the highly unstable 1-hydroxymethylbilane. The 1-hydroxymethylbilane is transformed into uroporphyrinogen III by uroporphyrinogen III synthase. When this enzyme is inactivated, 1-hydroxymethylbilane cyclizes non-enzymatically to uroporphyrinogen I, which can be oxidized to uroporphyrin I. PBGD activity can be measured by quantitation of uroporphyrin I formed from PBG under conditions where this is the only end product. The purpose of the present study was to define the optimal conditions for quantitating PBGD activity in human erythrocytes. The preanalytical factors examined were: anticoagulants and methods for disruption of the erythrocytes. The analytical factors examined were: duration of preincubation, reaction time, reaction temperature, pH, ionic strength and conditions for the oxidation of uroporphyrinogen I to uroporphyrin I. Based on the results, we propose an optimized method for determination of PBGD activity in erythrocytes.

Porphobilinogen deaminase (PBGD) is an enzyme involved in the synthesis of heme. Acute intermittent porphyria (AIP) is an inherited disease resulting from a reduced activity of PBGD. The symptoms seem to be due to a neurological dysfunction. Attacks of AIP are often provoked by conditions where the PBGD activity becomes insufficient as a result of an increased synthesis of heme in the liver. How this affects the nervous tissue is still unknown. It may well be that a reduced activity of PBGD in other tissues than the liver is of importance too. The aim of the present study was to examine the activity and the immunohistochemical localization of PBGD in the following tissues of wistar female rats: brain, heart, submandibular gland, liver, kidney, pancreas, ovary, stomach, duodenum, jejunum, ileum, colon and musculature. The PBGD activity varied considerably among the tissues. It was highest in the liver, 14 pkat/g, and lowest in the jejunum, 0.7 pkat/g. The immunohistochemical localization of PBGD was studied by antibodies raised against a 40 amino acid synthetic peptide that corresponds to a segment in the C-terminal part of PBGD. The study demonstrated that the PBGD immunoreactivity was not evenly distributed among the various cell types in a given tissue. Immunohistochemical reactions were pronounced in Kupffer cells in the liver, in smooth muscle cells of arteries and arterioles, in distal and collecting tubules in the kidney, in nerve axons in the brain and in ganglionic cells in the intestine. Especially, the immunohistochemical reaction in nerve cells is notable considering the nervous dysfunction in AIP.

Background: Cystatin C is a proteinase inhibitor with a low molecular weight. The serum levels of cystatin C are mainly dependent on glomerular filtration rate (GFR) making cystatin C an endogenous parameter of GFR. The aim of the study was to elucidate the applicability of serum cystatin C as a parameter of GFR in patients with normal to moderately impaired kidney function and to estimate a reference interval for serum cystatin C., Patients and Methods: Forty-six patients (25 males and 21 females) aged 22 to 83 years with various kidney diseases and 250 blood donors (164 males and 86 females) aged 19 to 64 years were included. Cystatin C was measured by an automated particle-enhanced nephelometric immunoassay, serum creatinine by an enzymatic and by Jaffé method, urine creatinine by an enzymatic method, and GFR by 99mTc-DTPA clearance., Results: Serum levels ofcystatin C and creatinine showed increments with decreasing values of 99mTc-DTPA clearance and a linear relationship was found between 99mTc-DTPA clearance and l/serum cystatin C, l/serum creatinine (enzymatic method), and creatinine clearance. Comparison of the non-parametric receiver-operating characteristic (ROC) plots for serum cystatin C (area under the curve (AUC) = 0.996; SE = 0.005), serum creatinine (enzymatic method) (AUC = 0.899; SE = 0.044), serum creatinine (Jaffé method) (AUC = 0.870; SE = 0.051), measured creatinine clearance (AUC = 0.959; SE = 0.025), and estimated creatinine clearance (0.950; SE = 0.029) revealed significant differences for serum cystatin C and serum creatinine (enzymatic and Jaffé method) (p values: 0.03 and 0.01). No significant differences were demonstrated between serum cystatin C and measured and estimated creatinine clearance (p value: 0.14 and 0.12). The non-parametric reference interval for serum cystatin C was calculated to be 0.51-1.02 mg/l (median: 0.79 mg/l; range: 0.33 - 1.07 mg/l)., Conclusion: Serum cystatin C seems to be a better parameter of GFR than serum creatinine in adults with various types of kidney disease with normal to moderately impaired kidney function.

Measurement of the serum concentration of the acute-phase reactant C-reactive protein (CRP) provides an useful objective marker in clinical practice, both in screening for diseases and in monitoring disease activity and response to therapy. The aim of this study was to evaluate the analytical performance of the assay in the concentration range from 0.2 to 15 mg/L, and to determine a reference interval for serum CRP in healthy blood donors using the Dade Behring N Latex CRP mono assay on the Dade Behring Nephelometer II. The assay is a particle-enhanced nephelometric immunoassay with a measuring range of 0.2-1100 mg/L. The sample volume is 40 microL, and the assay time is 6 min. The coefficients of variation for within-run and between-run imprecision studies were between 2.2 and 5.4% in serum pools with CRP concentrations between 1.29 and 11.98 mg/L. Linearity studies showed excellent correlation between the theoretical and the measured values. No interferences were detected from hemoglobin <1.0 mmol/L, bilirubin <512 micromol/L and Intralipid <20 g/L. Blood samples were collected from 268 healthy blood donors (107 females and 161 males) between 20 and 65 years old. The non-parametric reference intervals for serum CRP was calculated to be <0.2-14.7 mg/L in females and <0.2-9.16 mg/L in males. The Mann-Whitney U-test revealed no gender-related difference for serum CRP (p-value=0.72). A common reference interval for serum CRP for both genders was calculated to be <0.2-10.5 mg/L (median 0.98 mg/L, range <0.2-17.3 mg/L).

Since 1985, cystatin C has been suggested to be a marker of the renal function. Cystatin C is a proteinase inhibitor with a low molecular weight (M(r) = 13359). It is produced at a constant rate in all nucleated cells investigated to date, freely filtered in the renal glomeruli and reabsorbed and catabolised in the proximal tubules. The concentration of serum cystatin C is mainly determined by glomerular filtration, which makes cystatin C an endogenous marker of glomerular filtration rate (GFR). There are few data describing the influence of various factors on the production and elimination of cystatin C. Fully automated assays using particle-enhanced turbidimetry or particle-enhanced nephelometry are available and the assays are precise, rapid and usable in clinical routine practice. Reference intervals have been determined for cystatin C in adults and in children older than one year. It has been suggested that the same reference interval can be used in children older than one year and in adults without gender differences, on the assumption that the same method with the same standardisation is used. Several studies including adults and children with different renal diseases with various kidney function have suggested serum cystatin C to be a better marker of GFR than serum creatinine.

The Dade Behring N Latex Cystatin C assay, a particle-enhanced nephelometric immunoassay for measuring serum cystatin C, was evaluated on the Dade Behring Nephelometer II. The assay time was 6 min and the throughput was 75 samples per hour. The sample volume was 40 microL and the measuring range was 0.25-7.90 mg/L. Imprecision studies revealed within-run CVs < 1.8% and between-run CVs < 1.8% in the concentration range 0.87-4.63 mg/L. Recovery was 92.4-101.3%. Linearity studies showed excellent correlation between the theoretical and obtained values. No interferences were detected from haemoglobin < 1.0 mmol/ L, bilirubin <512 micromol/L and Intralipid <20 g/L. Stability of cystatin C in serum was 7 days at temperatures from 20 degrees C to 20 degrees C and 6 months at -80 degrees C. Measurements of cystatin C in heparin-plasma and EDTA-plasma did not differ significantly from cystatin C measured in serum. Fifty patient samples run on the Dade Behring Nephelometer II (y) were compared to the Dako Cystatin C assay (x). The Passing-Bablok regression analysis revealed y = 1.105x - 0.340. In conclusion, the Dade Behring N Latex Cystatin C assay was precise and correlated with the Dako Cystatin C assay.

The protease inhibitor cystatin C is a non-glycosylated low molecular weight protein (Mr=13359) which is produced by all nucleated cells at a constant rate, freely filtered by the renal glomeruli, and catabolized in the tubuli. The aim of the study was to elucidate the applicability of serum cystatin C as a marker of glomerular filtration rate (GFR) in patients with various kidney diseases with a wide range of renal function and in dialysis patients. Seventy-six patients with various kidney diseases (aged 20 to 79 years) and 61 dialysis patients (aged 21 to 82 years) were included. Serum cystatin C was measured by automated particle-enhanced immunoturbidimetry, serum and urine creatinine by an enzymatic method, and GFR by 99mTc-DTPA-clearance using a single plasma sample method. Serum cystatin C in patients with various kidney diseases was 1.90+/-0.98 mg/L (mean+/-SD) and in dialysis patients 7.14+/-1.91 mg/L. In the non-dialysis patients a linear relationship was found between 99mTc-DTPA-clearance and 1/serum cystatin C (r=0.91, p-value<0.0001), 1/serum creatinine (r=0.89, p-value<0.0001), and creatinine-clearance (r=0.88, p-value<0.0001). Comparison of the non-parametric ROC plots for serum cystatin C (area under the curve (AUC)=0.9665; SE=0.0169), serum creatinine (AUC=0.9554; SE=0.0205), and creatinine-clearance (AUC=0.9731; SE=0.0160) revealed no significant differences (p-values: 0.50, 0.78, and 0.49). In conclusion, cystatin C may be a likewise good marker of the GFR as serum creatinine and creatinine-clearance, cystatin C having the advantage being independent of gender and muscle mass.

The aim of this study was to establish reference intervals for cerum cystatin C and serum creatinine in adults. Blood samples were collected from 270 healthy blood donors (135 men and 135 women between 20 and 65 years old with 15 men and 15 women in each five-year-interval). Serum cystatin C was analyzed using an automated particle-enhanced immunoassay (DAKO Cystatin C PET kit) on the Cobas Mira S analyzer. Serum creatinine was analyzed using the Vitros Creatinine Slide, an enzymatic method on the Vitros 950 chemistry analyzer. The calculated reference intervals for serum cystatin C were 0.62-1.15 mg/l in women (median 0.84 mg/l, range 0.56-1.29 mg/l) and 0.51-1.25 mg/l in men (median 0.87 mg/l, range 0.42-1.39 mg/l). The Mann-Whithey U-test revealed no gender-related difference for cystatin C (p = 0.48). A common reference interval in women and men was calculated to be 0.54-1.21 mg/l (median 0.85 mg/l, range 0.42-1.39 mg/l). The non-parametric reference interval for serum creatinine was 57-95 mumol/l in women (median 72 mumol/l, range 44-105 mumol/l) and 69-111 mumol/l in men (median 89 mumol/l, range 58-123 mumol/l).

Imipramine elimination and metabolism was studied in a single-pass rat liver perfusion model with flow and pressure kept constant for a perfusion-period of 120-200 min. Imipramine was eliminated from the liver mainly in three ways: unchanged in the outflow medium, by demethylation forming desipramine, and by 2-hydroxylation and subsequent glucuronide coupling. The formed desipramine was eliminated unchanged (outflow) or by 2-hydroxylation + glucuronide coupling. The conjugated metabolites were mainly eliminated via the bile. By perfusion with constant imipramine concentration in the inflow medium (10-15 microM), steady state could not be obtained within 120 min. perfusion time, neither in terms of a constant outflow concentration, nor in terms of balance between input and output. Perfusion with imipramine for 20 min. (single-dose) followed by perfusion with drug-free medium showed an extensive elimination of imipramine (extraction ratio 0.64-0.97) and rapid disappearance of imipramine from outflow medium (t 1/2 5-10 min). A second single dose imipramine perfusion given 70 to 90 min. after start showed a significant reduction in extraction ratio. At the same time the elimination via bile decreased about 50%, and only a fraction of this decrease was accounted for by a decrease in bile flow. The data thus indicate that a gradual decline in elimination rate may explain the difficulties in reaching a steady state during constant infusion with imipramine. Gradual changes in distribution kinetics could not be excluded though. Results of both types of experiments (continuous or single dose administration) indicated dose dependent elimination compatible with saturation kinetics.