Soluble transferrin receptor in urine,a new biomarker for IgA nephropathy and Henoch–Schönlein purpura nephritis

ObjectivesIgA nephropathy (IgAN) and Henoch–Schönlein purpura nephritis (HSPN) might represent different ends of a continuous spectrum of glomerular disease. In both conditions, upregulated soluble transferrin receptor (sTfR) might be excreted in urine, which could be a potential biomarker to monitor disease activity and therapeutic response.MethodsIn this pilot study, 132 Caucasian patients consulting the Nephrology Department at the Ghent University Hospital because of a glomerulopathy and 50 normal controls were included. Urinary sTfR concentrations were determined in concentrated urine using a newly developed latex-enhanced immunonephelometric assay.ResultsMedian urinary sTfR concentration was higher in patients with a primary glomerulopathy than in healthy subjects (p < 0.0001). More importantly, absolute median levels of urinary sTfR were markedly higher in patients with active IgAN or HSPN [10 μg/L, 95% confidence interval (CI): 6–18 μg/L] in comparison with those with other morphological types of glomerulopathy (2 μg/L, 95%CI: 1–4 μg/L) (p < 0.0001). A statistically significant difference in urinary sTfR concentration was observed between patients with active IgAN or HSPN and patients who had achieved partial or complete remission (p < 0.0001). Multiple regression analysis with urinary sTfR as dependent variable revealed that proteinuria was the main predictor of urinary sTfR concentration (r² = 0.52, p < 0.001).ConclusionDetermination of sTfR in urine is a new and sensitive method for a potential biomarker of IgAN and HSPN.     

Validating urinary measurement of beta-2-microglobulin with a Roche reagent kit designed for serum measurements

ObjectiveTo validate the use of a Roche serum beta-2-microglobulin (B2MG) kit for urinary B2MG measurements, and to establish reference limits for urinary B2MG/creatinine ratio from healthy individuals.Design and methodsThe Roche B2MG Tina-Quant serum kit was used to measure urinary B2MG immunoturbidimetrically.ResultsUsing human urine as a diluent, the B2MG method was linear from 73–2156 μg/L. The imprecision on a commercially available urine QC was 4.4% at a concentration of 380 μg/L. Limit of quantification at < 20% CV was 40 μg/L. Method comparison with Immulite 2000 (Siemens) yielded slope = 1.180 (95% CI 1.14–1.22), intercept = 11.5 (95% CI ? 3.6–26.6), SEE = 27.6 and r = 0.99 (n = 26) by the Deming regression analysis. The upper reference limit of B2MG/creatinine ratio determined from 195 healthy adults was 29 μg/mmol (97.5th centile).ConclusionsThe serum B2MG Tina Quant reagent kit is acceptable to measure urinary B2MG.     

Pro-HEART — A randomized clinical trial to test the effectiveness of a high protein diet targeting obese individuals with heart failure: Rationale,design and baseline characteristics

There is ample research to support the potential benefits of a high protein diet on clinical outcomes in overweight/obese, diabetic subjects. However, nutritional management of overweight/obese individuals with heart failure (HF) and type 2 diabetes mellitus (DM) or metabolic syndrome (MS) is poorly understood and few clinical guidelines related to nutritional approaches exist for this subgroup. This article describes the design, methods, and baseline characteristics of study participants enrolled in Pro-HEART, a randomized clinical trial to determine the short term and long term effects of a high protein diet (30% protein [~ 110 g/day], 40% carbohydrates [150 g/day], 30% fat [~ 50 g/day]) versus a standard protein diet (15% protein [~ 55 g/day], 55% carbohydrates [~ 200 g/day], 30% fat [~ 50 g/day]) on body weight and adiposity, cardiac structure and function, functional status, lipid profile, glycemic control, and quality of life. Between August, 2009 and May, 2013, 61 individuals agreed to participate in the study; 52 (85%) – mean age 58.2 ± 9.8 years; 15.4% Blacks; 57.7% Whites; 19.2% Hispanics; 7.7% Asians; 73.1% male; weight 112.0 ± 22.6 kg – were randomized to a 3-month intensive weight management program of either a high protein or standard protein diet; data were collected at baseline, 3 months, and 15 months. This study has the potential to reveal significant details about the role of macronutrients in weight management of overweight/obese individuals with HF and DM or MS.     

Circulating sTWEAK improves the prediction of coronary artery disease

ObjectivesDecreased concentrations of circulating soluble tumor necrosis factor-like weak inducer of apoptosis (sTWEAK) were recently reported to be associated with atherosclerosis, but there are still no data concerning its predictive performance.Design and methodsThe current cross-sectional study investigates the potential of the novel atherosclerotic biomarker for the prediction of coronary artery disease (CAD). Serum sTWEAK was measured by ELISA in 76 CAD patients and 82 CAD-free subjects.ResultsSerum sTWEAK concentrations were significantly lower in the patients (534.5 ± 110.9 μg/L) than in the controls (688.1 ± 150.0 μg/L, p < 0.001), even after adjusting for different confounders (p < 0.001). The areas under ROC curves (AUC)s calculated for logistic regression models that included different known risk factors were significantly increased when sTWEAK was added to the corresponding model (p = 0.011–0.035).ConclusionsThe measurement of serum sTWEAK concentrations improves the prediction of CAD based on existing biomarkers.     

A novel ultra-sensitive enzyme immunoassay for soluble human insulin receptor ectodomain and its measurement in urine from healthy subjects and patients with diabetes mellitus

ObjectiveFor the early identification of patients at risk of developing diabetes mellitus, and to prevent the onset of diabetes by performing dietary counseling and exercise guidance, we have developed an ultra-sensitive immune complex transfer enzyme immunoassay (ICT-EIA) to measure soluble human insulin receptor ectodomain (sIRα) in urine which is collected non-invasively.Design and methodsWe developed ICT-EIA for sIRα and measured urinary sIRα from 106 healthy volunteers, 35 obese volunteers and 42 patients with diabetes.ResultsThe detection limit of ICT-EIA (0.04 pg/mL), using a urine sample of as little as 100 μL, was a few hundred-fold higher than that of conventional ELISA. Using ICT-EIA, the urinary sIRα level in patients with diabetes (9.7 ± 20.1 pg/mg creatinine) was significantly higher than those in healthy volunteers (1.4 ± 0.9; P < 0.001).ConclusionICT-EIA for sIRα may be useful as a good marker for evaluating diabetes risk.     

Comparison of the effects of lidocaine pre-administration and local warming of the intravenous access site on propofol injection pain: Randomized,double-blind controlled trial

BackgroundLidocaine reduces pain that occurs upon the intravenous injection of propofol. But, there are few non-pharmacological nursing interventions to reduce propofol injection pain.ObjectiveTo compare the effects of lidocaine pre-administration and local warming of the intravenous access site on propofol injection pain.DesignProspective, double-blind, randomized controlled trial.SettingThe 555 bed, non-teaching National Cancer Center in Kyunggido, South Korea.ParticipantsA total of 96 patients who underwent thyroidectomy under total intravenous general anesthesia with propofol were randomly allocated to the control, lidocaine pre-administration (LA) or local warming (LW) group.MethodsAll three groups received 2% propofol with an effect-site target at 3 μg/mL for induction dose. The control group received 2% propofol with no intervention. The lidocaine pre-administration group received 2% propofol 30 s after 1% lidocaine 30 mg. The local warming group received 2% propofol after warming of the intravenous access site for 1 min using 43 °C forced air. Propofol injection pain was assessed by four-point verbal categorial scoring (VCS), numerical rating scale (NRS) and surgical pleth index (SPI).ResultsPain VCS of the LA group (mean ± SD, 1.11 ± 0.45) was significantly reduced (U = −3.92, p < .001) compared to the control group (mean ± SD, 1.71 ± 0.74). Pain VCS of the LW group (mean ± SD, 0.76 ± 0.44) was significantly reduced (U = −5.17, p < .001) compared to the control group (mean ± SD, 1.71 ± 0.74). Pain VCS of the LW group was significantly reduced compared to the LA group (U = −3.33, p = .001]. Pain NRS of the LA group (mean ± SD, 4.31 ± 2.32) was significantly reduced (mean difference, 1.82; 95% CI, 0.63–3.00; p = .003) compared to the control group (mean ± SD, 6.13 ± 2.39). Pain NRS of the LW group (mean ± SD, 3.06 ± 2.37) was significantly reduced (mean difference, 3.07; 95% CI, 1.63–4.51; p < .009) compared to the control group. There were significant differences in pain NRS between the LA group and the LW group (mean difference, 1.25; 95% CI, 0.09–2.42; p = .035). SPI of the LA group (mean ± SD, 64.1 ± 16.3) was significantly reduced (mean difference control versus LA, 8.36; 95% CI, 1.64–15.1; p = .016) compared to the control group (mean ± SD, 72.5 ± 9.56). SPI of the LW group (mean ± SD, 55.0 ± 16.2) was significantly reduced (mean difference control versus LW, 17.4; 95% CI, 10.8–24.0; p < .001) compared to the control group. There was a significant difference in SPI between the LA group and LW group (mean difference, 9.06; 95% CI, 1.02–17.1; p = .028).ConclusionLocal warming of the intravenous access site by 43 °C forced air for 1 min is slightly more effective in reducing propofol injection pain compared to lidocaine pre-administration.     

The Role of a Low-Dose Ketamine-Midazolam Regimen in the Management of Severe Painful Crisis in Patients With Sickle Cell Disease

ContextAcute pain is one of the main causes of hospital admission in sickle cell disease, with variable intensity and unpredictable onset and duration.ObjectivesWe studied the role of a low-dose intravenous (IV) ketamine-midazolam combination in the management of severe painful sickle cell crisis.MethodsA retrospective analysis was performed with data from nine adult patients who were admitted to the intensive care unit with severe painful sickle cell crises not responding to high doses of IV morphine and other adjuvant analgesics. A ketamine-midazolam regimen was added to the ongoing opioids as an initial bolus of ketamine 0.25 mg/kg, followed by infusion of 0.2–0.25 mg/kg/h. A midazolam bolus of 1 mg followed by infusion of 0.5–1 mg/h was added to reduce ketamine emergence reactions. Reduction in morphine daily requirements and improvement in pain scores were the determinants of ketamine-midazolam effect. The t-tests were used for statistical analysis.ResultsNine patients were assessed, with mean age of 27 ± 11 years. Morphine requirement was significantly lower after adding the IV ketamine-midazolam regimen. The mean ± SD IV morphine requirement (milligram/day) in the pre-ketamine day (D0) was 145.6 ± 16.5, and it was 112 ± 12.2 on Day 1 (D1) of ketamine treatment (P = 0.007). The Numeric Rating Scale scores on D0 ranged from eight to ten (mean 9.1), but improved to range from five to seven (mean 5.7) on D1. There was a significant improvement in pain scores after adding ketamine-midazolam regimen (P = 0.01).ConclusionLow-dose ketamine-midazolam IV infusion might be effective in reducing pain and opioid requirements in patients with sickle cell disease with severe painful crisis. Further controlled studies are required to prove this effect.     

Development of an enzymatic assay for sphingomyelin with rapid and automatable performances: Analysis in healthy subjects and coronary heart disease patients

BackgroundSphingomyelin (SM) is an important choline group-containing phospholipid and is considered to be an independent risk factor for coronary heart disease.MethodsWe have developed a specific enzymatic assay for SM measurement with rapid and automatable performances by using two-reagent system involving sphingomyelinase. We performed within-run and between-run precision, linearity test, detection limit, recovery test and interference to validate this assay. Then, we measured the serum SM concentration in 194 healthy subjects and 141 consecutive patients undergoing coronary angiography.ResultsThe within-run and between-run coefficients of variation for SM concentrations were 1.1–1.3% and 1.0–1.2%, respectively. Quantitative measurements to a lower limit of 30 μmol/L were shown to be possible. The recoveries of the exogenously added SM to the control samples were 98.7%–101.5%. No effect was observed after the addition of some interference materials. The mean ± SD of the serum SM concentration in the 194 healthy subjects was 553.3 ± 100.1 μmol/L. We found that the SM concentration was significantly higher among an acute coronary syndrome subjects than among the healthy subjects (P < 0.01) and that the serum SM concentrations were significantly correlated with the serum magnesium concentration.ConclusionsWe have developed a rapid and automatable enzymatic assay for SM that enables the automatic measurement of choline-containing phospholipids. This assay may be useful for various types of biochemical and clinical research.     

Twenty-four-hour profiles of urinary excretion of calcium,magnesium, phosphorus,urea, and creatinine in healthy prepubertal boys

ObjectivesAvailable data on 24-h urinary solute excretion in healthy children are sparse. We thus documented the daily and overnight variations of urinary electrolytes (calcium, magnesium, and phosphorus), urea, and creatinine in prepubertal (Tanner stage I) boys.Design and methodsNine voluntary healthy prepubertal boys aged 10.8 ± 0.11 years participated in this study. Concentrations of variables were quantified in daytime samples (collected between 07:00 h ± 30 min and 21:00 h ± 30 min) and nighttime samples (collected between 21:00 h ± 30 min and 07:00 h ± 30 min) in spring, during a period of 24-h every 3 h.ResultsSignificant differences were found between daytime and nighttime excretion of calcium (p < 0.05), magnesium (p < 0.001), phosphorus (p < 0.01), and urea (p < 0.05), with high concentrations during the night. The 24-h solute/creatinine ratio was 0.072 ± 0.008 mg/mg for calcium, 0.069 ± 0.008 mg/mg for magnesium, 0.698 ± 0.070 mg/mg for phosphorus, and 0.017 ± 0.001 g/mg for urea. Statistically significant correlation analyses showed that urea and creatinine were positively associated with body mass index (BMI) (R = 0.790, p = 0.0113 for urea; R = 0.889, p = <0.0013 for creatinine) and weight (R = 0.717, p = 0.0297 for urea; R = 0.978, p = < 0.001 for creatinine). The other urinary variables were independent of BMI and body mass.ConclusionThese data are of interest for the diagnosis of certain renal disease in prepubertal children.     

Urinary melamine and adult urolithiasis in Taiwan

BackgroundThis study aims to investigate the association between urinary melamine concentration and the risk of urolithiasis in adult.MethodsFrom 2003 to 2007, 11 and 22 patients diagnosed with upper urinary tract uric acid urolithiasis and calcium urolithiasis, respectively, were recruited from Kaohsiung Medical University Hospital. For comparison, we randomly collected 22 sex- and age-matched subjects who come to the same hospital for regular health check-up at the same period of time. Urinary melamine concentration was measured by the method of triple-quadrupole liquid chromatography tandem mass spectrometry (LC-MS/MS). Wilcoxon rank sum test was used to compare the urinary melamine concentrations in uric acid urolithiasis patients with controls as well as in calcium urolithiasis patients with controls. FDR (false discovery rate) was used to correct the p-values for two comparisons.ResultsSubjects with uric acid urolithiasis (median: 0.50 vs 0.06 μg/mmol creatinine, Wilcoxon test: FDR_p = 0.024) and with calcium urolithiasis (median: 0.14 vs 0.06, FDR_p = 0.024) had significantly higher urinary melamine concentration than controls. Based on the ROC curves, subjects whose melamine levels were over 0.262 and 0.037 μg/mmol creatinine, respectively, might have significant risks to have uric acid and calcium urolithiasis.ConclusionThis preliminary study suggests that exposure to even low-dose melamine-related products still have the potential to develop both uric acid and calcium urolithiasis in adults.     

Evaluation of Hemo Techt NS-Plus system for use in a province-wide colorectal cancer screening program

ObjectivesThe NS-Plus automated analyzer and fecal immunochemical testing (FIT) testing system (Alfresa Pharma) was evaluated for use in Newfoundland and Labrador's provincial colorectal cancer (CRC) screening program.Design and methodsVarious method performance characteristics were evaluated including the sample stability. The sensitivity for detecting neoplastic lesions was evaluated in 249 patients scheduled for colonoscopy. Each patient collected up to 2 samples for both guaiac based testing (Hemoccult ® SENSA; gFOBT) and FIT using the NS-plus system (cutoff = 20 μg Hb/g feces or 100 μg Hb/L) over 2 days. Data was analyzed comparing 1- and 2-day testing strategies.ResultsThe analyzer showed acceptable linearity, precision, and accuracy. The collection device maintained acceptable sample stability for at least 7 days at: 37 °C, room temperature (~ 23 °C), 4–8 °C, and ? 20 °C. The 2-day sampling strategy identified 30% (21 of 69) of all neoplastic lesions (low and high grade adenomas and CRC) including 2 of 4 high-grade adenomas and 2 of 2 CRCs. The single day strategy identified the same high-grade adenomas and CRCs but fewer low-grade adenomas (23% of all neoplasia). Reducing the screening cutoff to the estimated 95th percentile of FIT results in the healthy adult population (10 μg Hb/g feces), detected all high-grade adenomas in the 2-day strategy.ConclusionsThe NS Plus automated analyzer system detects clinically significant neoplasms and shows acceptable performance for use in a CRC screening program with the potential for gains in sensitivity by modifying the number of days of screening or through lowering the cutoff.     

Reference range of thyroid function (FT3, FT4 and TSH) among Indian adults

ObjectivesTo generate thyroid hormone reference norms using electro-chemiluminescence technique.Design and methodsCross sectional study on apparently normal 4349 Delhi adults (18–86 years). Predetermined exclusion criteria (goiter, hypoechogenicity or nodularity on ultrasound, elevated anti-thyroid peroxidase antibody, hypo or hyperthyroidism and family history of thyroid dysfunction) excluded 2433 subjects leaving 1916 (916 males and 1000 females) as the reference population.ResultsMean age and BMI of the reference population were 41.2 ± 18.1 years and 24.5 ± 4.4 kg/m² respectively. Median urinary iodine excretion was 233.6 μg/L (79–458;3rd–97th centile). The population was categorized into various age groups (18–30, 31–40, 41–50, 51–60, 61–70 and ≥ 70 years). Overall FT3 and FT4 values in the reference population irrespective of age, ranged from 2.4–8.8 (mean 4.6 ± 0.9) pmol/L and 10.1–24.8 (mean 15.40 ± 2.0) pmol/L, respectively. Mean TSH value in the reference population was 2.2 ± 0.9 mIU/L which was significantly lower than that of total population (3.8 ± 6.1; p < 0.001).ConclusionFT3 values were observed to be significantly higher in men than women (p = 0.001). The centiles (3rd, 5th, 10th, 25th, 50th, 75th, 90th, 95th and 97th) of FT3, FT4 and TSH were derived for reference purposes in Indian adults. This community based study in Indian adults has established mean reference intervals for FT3, FT4 and TSH for different age groups for both sexes separately using strict exclusion criteria. These can be used as reference norms for Indian adults.     

Alteration of HDL functionality and PON1 activities in acute coronary syndrome patients

ObjectiveThe functionality of HDL has been suggested as an important factor in the prevention of cardiovascular and coronary artery diseases. The objective of the present study was to investigate the functionality of HDL and the factors that may affect the anti-atherogenic properties of HDL in ACS patients.Methods and resultsOne hundred healthy subjects and 205 ACS patients were recruited. HDL functionality was evaluated by measuring their capacity to mediate cholesterol efflux from J774 macrophages. Oxidative stress status was determined by measuring plasma malondialdehyde (MDA), protein carbonyl, and vitamin E levels by HPLC. The PON1 Q192R polymorphism status and PON1 paraoxonase and arylesterase activities of the healthy subjects and ACS patients were also determined. The HDL of ACS patients displayed a limited capacity to mediate cholesterol efflux, especially via the ABCA1-pathway. MDA (7.06 ± 0.29 μM) and protein carbonyl (9.29 ± 0.26 μM) levels were significantly higher in ACS patients than in healthy subjects (2.29 ± 0.21 μM and 3.07 ± 0.17 μM, respectively, p < 0.0001), while α- and γ-tocopherol (vitamin E) levels in ACS patients were 8-fold (p < 0.001) and 2-fold (p < 0.05) lower than in healthy subjects. Paraoxonase, arylesterase and HDL-corrected PON1 activities (PON1 activity/HDL ratio) were significantly lower in ACS patients. Logistic regression analyses showed that high PON1 paraoxonase and arylesterase activities had a significant protective effect (OR = 0.413, CI 0.289–0.590, p < 0.001; OR = 0.232 CI 0.107–0.499, p < 0.001, respectively) even when adjusted for HDL level, age, BMI, and PON1 polymorphism.ConclusionThe results of the present study showed that the functionality of HDL is impaired in ACS patients and that the impairment may be due to oxidative stress and an alteration of PON1 activities.     

Serum adipocyte fatty acid binding proteins and adiponectin in patients with coronary artery disease: The significance of A-FABP/adiponectin ratio

BackgroundAdipocyte fatty acid binding protein (A-FABP) and adiponectin have been shown to play important roles in atherosclerosis. We investigated serum A-FABP, adiponectin and A-FABP/adiponectin ratio in patients with coronary artery disease (CAD).MethodsA total of 340 subjects who underwent coronary angiography (CAG) were classified into CAD group (n = 211) and non-CAD group (n = 129) according to the CAG. Serum A-FABP and adiponectin concentrations were determined by enzyme-linked immunosorbent assays.ResultsCAD patients tend to have higher A-FABP concentrations than non-CAD subjects, the difference is significant only between female CAD patients and controls [22.8 (18.6–25.7) ng/ml vs 18.1 (15.6–21.8) ng/ml, P = 0.008]. Serum A-FABP concentration was independently associated with Gensini scores in female subjects (P = 0.018). CAD patients have significant higher serum A-FABP/adiponectin ratio [1.51 ± 0.05 vs 0.89 ± 0.03 ng/μg, P < 0.01] than controls in both genders.ConclusionsSerum A-FABP is associated with CAD more closely in female than in male. The A-FABP/adiponectin ratio may be a more useful indicator for CAD than A-FABP or adiponectin alone.     

Heart-type fatty acid binding protein is an early marker of myocardial damage after radiofrequency catheter ablation

ObjectivesRadiofrequency (RF) ablation of arrhythmias induces myocardial damage and release of biomarkers. This study aimed to assess the kinetics of heart-type fatty acid-binding protein (h-FABP), a cytosolic protein released after myocardial injury incurred by both atrial and ventricular RF ablation, compared to other markers of myocardial injury.Design and methodsh-FABP, cTnI, CK-MB_mass and myoglobin were evaluated in 30 patients with atrial or ventricular tachyarrhythmias before, immediately after and at 3, 6 and 24 h after the procedure.Resultsh-FABP increased immediately after the procedure in all subjects (6.6 ± 1.2 μg/L vs 2.7 ± 0.3, p < 0.001) but increased significantly only in ventricular ablations. The peak of h-FABP significantly correlates with the values of time for mean power of RF application in both the entire patient cohort and in ventricular ablations.Conclusionsh-FABP may be an early parameter for monitoring RF-induced lesions and the site of ablation was relevant for biomarker increase.     

Simultaneous determination of sildenafil citrate and some nitric oxide releasing drugs in human plasma using UPLC MS/MS

ObjectiveThe inadvertent combination of sildenafil (SLD) and nitric oxide releasing compounds (NRC) may cause a life threatening hypotension and conversion of coital angina into an irreversible one. The aim of this study was to develop and validate a UPLC MS/MS method for the simultaneous quantitative analysis of SLD, nicorandil (NRD), and ARG in human plasma to determine the safety margins for drug combinations.Design and methodChromatographic elution was achieved in 4 min using gradient elution and an injection volume of 10 μL. Electro-spray positive ionization (ESI+ve) detection and multiple-reaction monitoring mode (MRM) were used for detection.ResultsThe method was found to be linear (10–900 ng/mL for SLD and NRD while 1–30 μg/mL for ARG), accurate and precise (99.35 ± 1.58, 99.62 ± 1.13, and 100.04 ± 1.22% for SLD, NRD and ARG; respectively) and met all other validation requirements.ConclusionThe developed UPLC MS/MS method is suitable for fast, sensitive, accurate and simultaneous determinations of SLD, NRD, and ARG in plasma.     

Changes in transferrin glycosylation during pregnancy may lead to false-positive carbohydrate-deficient transferrin (CDT) results in testing for riskful alcohol consumption

BackgroundAn alcohol-induced change in serum transferrin glycosylation, termed carbohydrate-deficient transferrin (CDT), is widely used as a biomarker of heavy long-term drinking. This study examined the transferrin glycosylation profile and the risk for false-positive CDT results during pregnancy.MethodsSerum samples were collected from 24 healthy pregnant women starting in gestation week 9–21, throughout pregnancy, and 8 or more weeks after delivery. Altogether 171 sera (5–9 samples/person) were analysed. Transferrin glycoforms were quantified as a percentage of total transferrin, using an HPLC candidate reference method for CDT.ResultsDuring pregnancy, the relative disialo-, pentasialo- and hexasialotransferrin levels increased gradually, whereas trisialo- and tetrasialotransferrin were reduced. This effect was most pronounced in the third trimester. For disialotransferrin, the main target in CDT testing, initial values of 1.07 ± 0.17% (mean ± SD) increased to 1.61 ± 0.23% before delivery (~ 50% increase). Nine (38%) pregnant women reached %disialotransferrin values ≥ 1.7% (97.5th percentile for controls) but all results were < 2.0%. In the postpartum samples, all glycoform levels had returned towards the starting values.ConclusionsThese results suggest that the cutoff for %disialotransferrin and %CDT employed to indicate heavy long-term drinking need to be raised slightly in pregnant women, to minimize the risk for false-positive results on CDT testing.     

Verification of an immunoturbidimetric assay for heart-type fatty acid-binding protein (H-FABP) on a clinical chemistry platform and establishment of the upper reference limit

BackgroundHeart-type fatty acid-binding protein (H-FABP) is an early biomarker of cardiac injury. Randox Laboratories developed an immunoturbidimetric H-FABP assay for non-proprietary automated clinical chemistry analysers that could be useful in the emergency department. We verified the analytical performances claimed by Randox Laboratories on Roche Cobas 6000 clinical chemistry platform in use in our laboratory, and we defined our own 99th percentile upper reference limit for H-FABP.MethodsFor the verification of method performances, we used pools of spared patient samples from routine and two levels of quality control material, while samples for the reference value study were collected from 545 blood donors. Following CLSI guidelines we verified limit of blank (LOB), limit of detection (LOD), limit of quantitation (LOQ), repeatability and within-laboratory precision, trueness, linearity, and the stability of H-FABP in EDTA over 24 h.Results and discussionThe LOQ (3.19 μg/L) was verified with a CV% of 10.4. The precision was verified for the low (mean 5.88 μg/L, CV = 6.7%), the medium (mean 45.28 μg/L, CV = 3.0%), and the high concentration (mean 88.81 μg/L, CV = 4.0%). The trueness was verified as well as the linearity over the indicated measurement interval of 0.747–120 μg/L. The H-FABP in EDTA samples is stable throughout 24 h both at room temperature and at 4 °C. The H-FABP 99th percentile upper reference limit for all subjects (3.60 μg/L, 95% CI 3.51–3.77) is more appropriate than gender-specific ones that are not statistically different.     

The role of hypercytokinemia in the pathophysiology of tumor lysis syndrome (TLS) and the treatment with continuous hemodiafiltration using a polymethylmethacrylate membrane hemofilter (PMMA-CHDF)

ObjectiveTo examine the role of hypercytokinemia in the pathophysiology of tumor lysis syndrome (TLS) and the efficacy of continuous hemodiafiltration in the treatment of TLS.Design and settingRetrospective observational study in a general intensive care unit of a university hospital.PatientsFour patients with hematological disorder developing TLS after the treatment of anti-tumor chemotherapy.InterventionsContinuous hemodiafiltration using a polymethylmethacrylate membrane hemofilter (PMMA-CHDF) was performed at the onset of TLS. Blood samples were collected daily after ICU admission, and clinical parameters and blood levels of cytokines were evaluated.Measurements and resultsAll four patients underwent induction anti-tumor chemotherapy, during which they developed hyperuricemia, hyperkalemia, and acute renal failure. Two of them also developed multiple organ failure. Serum levels of tumor necrosis factor (TNF) -alpha, interleukin-6 (IL-6), and IL-10 prior to the initiation of PMMA-CHDF were 102 ± 85 pg/mL, 1097 ± 546 pg/mL, and 98 ± 83 pg/mL, respectively (mean ± SD). After three days of PMMA-CHDF treatment, corresponding blood levels were 37 ± 55 pg/mL, 326 ± 511 pg/mL, and 9 ± 8 pg/mL, respectively. Thus, all cytokine levels were significantly decreased by three days of PMMA-CHDF treatment (p < 0.05, paired t-test). Following three days of PMMA-CHDF treatment, blood urea nitrogen (BUN) and serum creatinine (Cre.) were significantly decreased (pre/post BUN 42.3 ± 15.4/16.5 ± 8.4 mg/dL, p < 0.05, pre/post Cre. 2.7 ± 1.2/1.2 ± 0.6 mg/dL, mean ± SD, p < 0.05). Furthermore, the clinical condition of each patient was improved after the treatment of PMMA-CHDF, and all of four patients were survived.ConclusionHypercytokinemia plays a pivotal role in the pathophysiology of TLS and PMMA-CHDF may be an effective therapeutic modality for TLS patients not only as renal replacement therapy but also as a cytokine modulator.     

Superiority of postmortem liver fentanyl concentrations over peripheral blood influenced by postmortem interval for determination of fentanyl toxicity

ObjectiveThe current study was undertaken to determine the relationship between postmortem (PM) peripheral blood (PB) and liver fentanyl concentrations and the role of measuring liver fentanyl concentrations in cause of death investigations in medical examiner cases in which fentanyl was identified.Design and methodsFB and liver tissue were routinely collected at autopsy from 4 Minnesota medical examiners' offices in 2010–2011. Samples were analyzed by gas chromatography–mass spectrometry (GC–MS).ResultsPB fentanyl ranged from < 2–15 μg/L in non-drug related deaths (n = 5), < 2–22 μg/L from mixed drug toxicity (n = 26) and 3.7–56 μg/L from fentanyl toxicity (n = 33). Liver fentanyl ranged from 11 to 104 μg/kg, 6 to 235 μg/kg, and 18 to 365 μg/kg, respectively. PB and liver fentanyl showed a modest correlation (r = 0.67). PM interval to the liver/blood ratio showed a decreasing ratio over increasing PM interval in cases from fentanyl and mixed drug toxicity. Liver fentanyl concentrations best define therapeutic use at < 23 μg/kg and fatal toxicity at > 56 μg/kg, without substantial overlap as found in blood fentanyl concentrations.ConclusionDiscriminatory liver fentanyl concentrations suggestive of therapeutic or toxic drug levels may better assist cause of death determination in cases of suspected fentanyl toxicity than postmortem PB concentrations. Peripheral blood fentanyl concentrations appear to undergo postmortem redistribution, associated with an increasing PM interval.