Severe rhabdomyolysis due to idiopathic inflammatory myopathies, a wary manifestation of a heterogenous pathology.

Idiopathic inflammatory myopathies (IIM) are historically classified by The Bohan and Peter criteria. The presentation of IIM is versatile and clinical-serological findings can aid in diagnosing the underlying form of IIM. Over the past years, the discovery and the use of myositis-specific autoantibodies (MSA) and myositis-associated autoantibodies (MAA) have led to a more heterogeneous classification by the European League Against Rheumatism and American College of Rheumatology (EULAR/ACR).This paper describes a case of dermatomyositis sine dermatitis. A 70-year -old woman presented with complaints of muscle weakness and was admitted because of severe oliguric renal failure due to rhabdomyolysis. Despite treatment with hemodialysis and initial recovery, her clinic worsened again. The disease course in combination with electromyography findings, PET-scan results, and positive myositis-specific autoantibodies, that is, anti-NXP-2 antibodies, ultimately led to the diagnosis.Today, commercial kits based on line immunoassay and dot blot have mostly replaced the labor-intensive immunoprecipitation of RNA and/or proteins for detecting MSA. Though it makes routine testing of multiple MSA easy to implement in clinical practice, more validation studies are required and clinicians should be aware of its limitations, including false-positive results. When clinical suspicion for IIM is high, a negative screening for antinuclear antibodies (ANA) result does not exclude IIM and the first test of choice remains a multi-specific immunoassay for the whole spectrum of MSA.In this paper, we want to underline that there is no shortcut in diagnosing IIM. Caution is required in interpreting different EMG, PET-scan, histological, and laboratory findings. Especially in the case of rhabdomyolysis, as this is a severe and wary manifestation of myositis.

Humoral Immune Response to SARS-CoV-2.

OBJECTIVES: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) serology tests are clinically useful to document prior SARS-CoV-2 infections. Data are urgently needed to select assays with optimal sensitivity at acceptable specificity for antibody detection. METHODS: A comparative evaluation was performed of 7 commercial SARS-CoV-2 serology assays on 171 sera from 135 subjects with polymerase chain reaction-confirmed SARS-CoV-2 infection (71 hospitalized patients and 64 paucisymptomatic individuals). Kinetics of IgA/IgM/IgG seroconversion to viral N and S protein epitopes were studied from 0 to 54 days after onset of symptoms. Cross-reactivity was verified on 57 prepandemic samples. RESULTS: Wantai SARS-COV-2 Ab ELISA and Orient Gene COVID-19 IgG/IgM Rapid Test showed superior overall sensitivity for detection of SARS-CoV-2 antibodies. Elecsys Anti-SARS-CoV-2 assay and EUROIMMUN Anti-SARS-CoV-2 combined IgG/IgA showed acceptable sensitivity (>95%) vs the consensus result of all assays from 10 days post onset of symptoms. Wantai SARS-COV-2 Ab ELISA, Elecsys Anti-SARS-CoV-2 assay, and Innovita 2019-nCoV Ab rapid test showed least cross-reactivity, resulting in an optimal analytical specificity greater than 98%. CONCLUSIONS: Wantai SARS-COV-2 Ab ELISA and Elecsys Anti-SARS-CoV-2 assays are suitable for sensitive and specific detection of SARS-CoV-2 antibodies from 10 days after onset of symptoms.

Disulfiram inhibition of cyanide formation after acetonitrile poisoning.

CONTEXT: Cyanide poisoning may be caused by acetonitrile, a common industrial organic solvent and laboratory agent. OBJECTIVE: To describe the potential use of disulfiram in treating acetonitrile poisoning in a human clinical case and to further study its effect in human liver microsomes in vitro. CASE DETAILS: A 30-year-old man initially presented with a cholinergic toxic syndrome following ingestion of aldicarb. Toxicological analysis revealed coingestion of ethanol. He subsequently developed severe metabolic acidosis caused by the cyanogenic compound acetonitrile which was erroneously interpreted as acetone in the chromatogram. After three treatments with hydroxocobalamin (5 g i.v.) and sodium thiosulfate (12.5 g i.v.) on days 2, 3, and 5, he had transient improvement but recurrent lactic acidosis. Treatment with disulfiram was associated on day 7 with resolution of metabolic acidosis and slowing of the decrease in acetonitrile concentration. He recovered from acetonitrile toxicity completely. The time course of acetonitrile, thiocyanate, and cyanide concentrations suggested that disulfiram inhibited cyanide formation. RESULTS: In vitro experiments with human liver microsomes showed the cyanide concentration was significantly lower after incubation with acetonitrile and disulfiram than acetonitrile alone (a mean 60% reduction in cyanide level). DISCUSSION: Although disulfiram was given late in the course of the poisoning it is possible that it contributed to the recovery.

Influence of physical properties of cuvette surface on measurement of serum lipase.

BACKGROUND: Despite striking similarities among colorimetric lipase assay recipes, marked intervendor differences are noted in the reported lipase values. In the present study, the effect of physical properties of the cuvette surface on measurement of serum lipase was investigated. METHODS: Lipase activity was measured concomitantly in cuvettes from three different analyzers: Vista (Siemens), Modular (Roche), and Synchron (Beckman Coulter). The surface/volume ratio of the cuvettes and the contact angle of the cuvette polymers were determined. The effects of various characteristics of serum (biochemical parameters, surface tension) were also examined. RESULTS: Serum lipase activities based on the colorimetric methylresorufin assay differed markedly according to the cuvettes used. More specifically, in the lower activity rate, marked differences were reported. The physical properties of the various cuvettes showed remarkable differences, especially in the contact angles. Other biochemical parameters (bilirubin, alkaline phosphatase, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and triglycerides) and serum surface tension did not affect the results. CONCLUSIONS: Serum lipase activity is affected by the physical properties of the cuvette surface.

A sensitive quantitative test strip based point-of-care albuminuria screening assay.

BACKGROUND: Chronic kidney disease is a major health problem and the global guidelines require screening of albuminuria. Therefore, affordable and sensitive albuminuria screening tests are needed. We explored the potential of urine strips, generally reported in the ordinal scale, measured on an automatic strip reader for reporting quantitative and sensitive albumin results. METHODS: We compared reflectance data of Combur-Test® strips obtained from the Cobas U411 reader (Roche) with albuminuria data from a nephelometer BNII (Siemens) and with protein concentrations from the pyrogallol red method (Modular P, Roche) for 389/328 non-pathologic and pathologic urine samples, respectively. RESULTS: Imprecision of the reflectance signal of the Cobas U411 was measured with commercial control material (Bio-Rad). Inter-run coefficients of variations (CVs) for reflectance for levels 1 and 2 were 1.7%/4.9%, respectively, and intra-run CVs were 1.8%/4.2%, respectively. Good agreement was obtained between the albumin concentration of the BNII and the protein strip reflectance data (n=389): Y (10,000/protein reflectance, 1/%)=160+0.132·X (albuminuria BNII, mg/L)-0.0000111·X2 (albuminuria BNII, mg/L); r2=0.921. Lower agreement was found between the protein assay (n=328) and the reflectance (r2=0.831). A calibration curve was made between 11.5 mg/L and 121.5 mg/L. The limit of blank (LOB) was 44.7 mg/L. CONCLUSIONS: The present study demonstrates that reflectance data generated by a test strip reader allows for quantitative analysis of albumin. Although the lower limit of the microalbumin range (30 mg/L) cannot be achieved with the dye-binding method, the results are satisfactory for screening purposes.

Mesothelin levels in urine are affected by glomerular leakage and tubular reabsorption.

BACKGROUND: Mesothelin is a soluble biomarker of malignant mesothelioma. Levels in serum, however, are also influenced by other factors, including age and glomerular filtration rate (GFR). The measurement of mesothelin in urine has recently gained interest, but the renal handling of this protein has not been sufficiently examined. PATIENTS AND METHODS: A total of 75 patients with benign kidney disease were prospectively included in the study. Mesothelin levels were measured in the serum and in the urine of all the participants by using enzyme-linked immunosorbent assay. Urinary albumin and alpha 1-microglobulin (A1M) levels, which are markers of glomerular leakage and of decreased tubular reabsorption, respectively, and the estimated GFR (eGFR) of each participant were obtained. All urine analyte levels were standardized (std) against urinary creatinine levels. RESULTS: Absolute mesothelin levels in urine (median, 0.58 nmol/L; interquartile range [IQR], 0.25-1.03 nmol/L) were significantly lower than those in serum (median, 1.74 nmol/L; IQR, 1.35-2.43 nmol/L; P < .001). Urinary mesothelin(std) levels positively correlated with serum mesothelin (r = 0.35, P < .01), albumin(std) (r = 0.51, P < .001), and A1M(std) levels (r = 0.71, P < .001). Neither age nor eGFR were associated with urinary mesothelin(std) levels. Similarly, multiple linear regression analysis indicated that only albumin(std) and A1M(std) levels were significantly positively associated with the urinary mesothelin(std) levels (adjusted R(2) = 0.49). CONCLUSION: Mesothelin levels in urine are affected by impaired glomerular and tubular function, which can influence the interpretation of mesothelin measurements and might cause false-positive results. These effects need to be accounted for to improve the further validation and possible clinical use of urinary mesothelin.

Performance of the Roche Total Mycophenolic Acid® assay on the Cobas Integra 400®, Cobas 6000® and comparison to LC-MS/MS in liver transplant patients.

BACKGROUND: Mycophenolic acid (MPA) is an immunosuppressant for which therapeutic drug monitoring (TDM) is performed for optimal prophylaxis and avoidance of toxicity in transplant patients. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is ideally suited for TDM of MPA. There have been several method comparisons of the Roche Total MPA assay, but none have been performed with respect to liver transplant patients. METHODS: We validated the Roche Total MPA assay on the Cobas Integra 400 and Cobas 6000 and compared it to a validated LC-MS/MS (API 2000™) method. Fifty-five EDTA plasma samples from liver transplant patients were measured with the Roche assay on these platforms and compared to the LC-MS/MS results. RESULTS: Validation of the LC-MS/MS, Cobas Integra 400 and 6000 was performed with good results. The LC-MS/MS/Integra 400/Cobas 6000 were linear up to 30, 15 and 17 mg/L, respectively. Imprecision was <10% for LC-MS/MS and <7% for the Roche assay on both platforms. The samples showed good agreement with LC-MS/MS. Passing-Bablok regression analysis showed Cobas Integra (mg/L)=1.02 × LC-MS/MS (mg/L)-0.50 and Cobas 6000 (mg/L)=0.98 × LC-MS/MS-0.47. CONCLUSIONS: The Roche Total Mycophenolic Acid-assay is suitable for measuring total MPA in plasma from liver transplant patients and is a good alternative for LC-MS/MS.

Evaluation of a new set of automated chemiluminescense assays for anticardiolipin and anti-beta2-glycoprotein I antibodies in the laboratory diagnosis of the antiphospholipid syndrome.

INTRODUCTION: The laboratory diagnosis of antiphospholipid syndrome (APS) requires the demonstration of antiphospholipid antibodies (aPL): lupus anticoagulant (LAC) measured through coagulation assays, anticardiolipin IgG or IgM antibodies (aCL) and/or anti-ß2glycoprotein I IgG or IgM antibodies (aß2GPI), usually detected by ELISA. MATERIALS AND METHODS: We evaluated the diagnostic value of aCL and aß2GPI measured by a new automated system using the chemiluminescence principle, the immunoanalyzer Zenit RA (Menarini). RESULTS: Results of aCL and aß2GPI were correlated with the clinical background of the patients and with results of ELISA (n=314). Correlated to the clinical background sensitivity/specificity ranged for aCL IgG between 7.5-45.2% / 54.2-98.8%, for aCL IgM 3.4-5.5% / 89.9-94%, for aß2GPI IgG 5.5-25.3% / 75.6-100% and aß2GPI IgM 3.4-4.8% / 89.9-92.3%, depending on the cut-off used. Sensitivity with manufacturer's cut-offs was comparable to ELISA, except for aß2GPI IgG with a significantly lower sensitivity compared to ELISA (5.5% vs 11.6%). In the APS patient population (n=30) sensitivity of aCL IgG and aß2GPI IgG was higher measured by ELISA compared to Zenit RA (46.7% vs 30.0%, and 46.7% vs 26.7%, respectively). Agreement between Zenit RA results and ELISA results for the four parameters was moderate (Kappa-values ranging 0.509-0.565). Sensitivity was 38.5%, 53.3%, 40% and 69.2% for aCL IgG, aCL IgM, aß2GPI IgG and aß2GPI IgM, respectively, applying the highest cut-off value for Zenit RA, raising towards 64.3%, 100%, 57.1%, for aCL IgG, aCL IgM, aß2GPI IgG, respectively, in a APS patient population. CONCLUSIONS: The new technology of chemiluminescense for measuring aPL showed good performance characteristics. Interpretation of results with a cut-off value associated with a good discrimination for disease, resulted in a lower sensitivity for the diagnosis of APS for aß2GPI IgG measured by Zenit RA assays compared to ELISA; sensitivity for aCL IgG was comparable to ELISA. Specificity for all parameters was high and comparable for aCL and aß2GPI.

Increased urinary neutrophil gelatinase associated lipocalin in urinary tract infections and leukocyturia.

BACKGROUND: Neutrophil gelatinase associated lipocalin (NGAL) is a protein present in neutrophils. NGAL is a promising biomarker for acute kidney injury. In urinary tract infections, urinary neutrophils can be a potential source of urinary NGAL. We investigated the effects of urinary tract infection and urinary neutrophil counts on urinary NGAL values. METHODS: NGAL was assayed using an immunoassay (ARCHITECT). Urine flowcytometry was performed with the UF-1000i (Sysmex). RESULTS: A correlation between the urinary white blood cell (WBC) count and NGAL concentrations was observed: log(Y) (NGAL, µg/L)=1.284+0.439 log(X) (urinary WBC, 10(9) cells/L); r=0.518. Similarly, the bacterial count correlated weakly with NGAL: log(Y) (NGAL, µg/L)=1.796+0.124 log(X) (bacterial count, 10(9) cells/L); r=0.243. Albuminuria correlated moderately with NGAL values: log(Y) (NGAL, µg/L)=1.557+0.339 log(X) (albuminuria, mg/L); r=0.368; α(1)-microglobulin (a1M) correlated weakly with NGAL: log(Y) (NGAL, µg/L)=1.631+0.360 log(X) (a1M, mg/L); r=0.381. CONCLUSIONS: Leukocyte contributions to urinary NGAL concentrations can be important. In leukocyturia or tubular damage (e.g., intensive care patients), using a fixed cut-off value for interpreting urinary NGAL data can lead to false positive results. Therefore, we suggest a mathematical correction in cases with pyuria (>100×10(9) cells/L) and urinary NGAL concentration >100 µg/L: corrected NGAL (µg/L)=NGAL-0.12 (urinary WBC, 10(9) cells/L).