Performance Evaluation of the Siemens SARS-CoV-2 Total Antibody and IgG Antibody Test.

OBJECTIVE: In this study, the performance of 2 commercially available SARS-CoV-2 antibody assays is evaluated. METHODS: The Siemens SARS-CoV-2 Total (COV2T) and IgG (COV2G) antibody tests were evaluated on a Siemens Atellica IM1300 analyzer. Imprecision was assessed with the CLSI EP15 protocol using positive controls. Ninety control group specimens were analyzed for specificity, and 175 specimens from 58 patients with polymerase chain reaction-confirmed SARS-CoV-2 were measured for the sensitivity and kinetics of the antibody response. RESULTS: Within-run and total imprecision were acceptable for both assays. Both tests showed a specificity of 100%. Sensitivity earlier in the disease state was greater for the COV2T assay than for the COV2G assay, but sensitivity >14 days after onset of symptoms approached 100% for both. For all patients, antibody titers remained above the seroconversion cutoff for all follow-up specimens. CONCLUSION: This study shows acceptable performance for both the Siemens COV2T and COV2G test, although seroconversion occurs earlier with the COV2T test.

Pseudonephritis is associated with high urinary osmolality and high specific gravity in adolescent soccer players.

The study aimed to evaluate the effect of exercise on urine sediment in adolescent soccer players. In 25 15-year-old (range 14.4-15.8 yrs) athletes, urinary protein, osmolality and cytology were analyzed by flow cytometry and automated dipstick analysis before (T(0)), during (T(1)), and after a match (T(2)). All athletes had normal urine analysis and blood pressure at rest, tested before the start of the soccer season. Fifty-eight samples were collected (T(0): 20, T(1): 17, T(2): 21). Proteinuria was present in 20 of 38 samples collected after exercise. Proteinuria was associated with increased urinary osmolality (p < .001) and specific gravity (p < .001). Hyaline and granular casts were present in respectively 8 of 38 and 8 of 38 of the urinary samples after exercise. The presence of casts was associated with urine protein concentration, osmolality, and specific gravity. This was also the case for hematuria (25 of 38) and leucocyturia (9 of 38). Squamous epithelial cells were excreted in equal amounts to white and red blood cells. A notable proportion of adolescent athletes developed sediment abnormalities, which were associated with urinary osmolality and specific gravity.

Maximum oxygen uptake at peak exercise in elderly patients with coronary artery disease and preserved left ventricular function: the role of inflammation on top of tissue Doppler-derived systolic and diastolic function.

BACKGROUND: Several studies have shown that longitudinal systolic function and left ventricular filling pressures, as assessed with tissue Doppler imaging, predict exercise capacity. AIM: The aim of this study was to evaluate whether natriuretic peptides and inflammatory parameters can independently predict maximum oxygen uptake at peak exercise (VO2max) on top of tissue Doppler imaging-derived markers. METHODS: We evaluated 142 patients (age 70 +/- 6 years, 77% men) with known or suspected coronary artery disease and a preserved left ventricular ejection fraction (> or = 50%). All patients underwent bicycle spiroergometry, and N-terminal pro-B-type natriuretic peptide levels were determined. Cytokines (IL-6 and soluble tumor necrosis factor receptors 1 and 2) and high-sensitivity C-reactive protein were measured as inflammatory markers. Tissue Doppler imaging was applied to evaluate peak long axis systolic velocities (Sm) and early mitral annulus velocities (E'). Ratio of early transmitral flow (E) to E' was assessed as marker of left ventricular filling. Analysis of variance, comparing VO2max quartiles, was used to determine univariate predictors and linear regression to determine multivariate VO2max predictors. RESULTS: Average VO2max was 18.5 +/- 5.7 mL/kg per minute (range 6-36.6). Compared with the highest quartile, patients with low VO2max were more frequently women (P < .0001). N-terminal pro-B-type natriuretic peptide and cytokine levels were significantly higher in the lower VO2max categories. Longitudinal myocardial velocities increased, and E/E' decreased along with increasing VO2max. In multivariate linear regression analysis, VO2max was independently predicted by sex, glucose, Sm, E/E', and cytokine levels. CONCLUSION: Maximum oxygen uptake at peak exercise in patients with known or suspected coronary artery disease and preserved systolic function was independently predicted by inflammatory makers on top of tissue Doppler-derived systolic and diastolic function.

Quantitative determination of vigabatrin and gabapentin in human serum by gas chromatography-mass spectrometry.

BACKGROUND: Published methods for routine clinical monitoring of vigabatrin and gabapentin are often very laborious. A simple GC-MS method was developed for the simultaneous quantitative determination of vigabatrin and gabapentin in human serum. METHODS: After protein precipitation, the compounds are derivatized by methylation and analysed on a polydimethylsiloxane column using splitless injection. Cyclobarbital is used as the internal standard. To attain maximal sensitivity, detection is performed in selected ion monitoring mode. RESULTS: The method was fully validated and linear calibration curves were obtained in the concentration ranges from 5 to 80 microg/mL for vigabatrin and from 5 to 30 microg/mL for gabapentin. The within-day and day-to-day relative standard deviations at three different concentration levels were <10% and <15%, respectively. The limit of quantitation was 2 mug/mL for both compounds. CONCLUSIONS: The presented method provides high chromatographic resolution, good sensitivity and unequivocal identification potential and can be used for simultaneous analysis of both antiepileptics.

Discrepancy between sperm acrosin activity and sperm morphology: significance for fertilization in vitro.

BACKGROUND: In routine semen analysis, discrepancies may occur between sperm acrosin activity test results and sperm acrosomal morphology. METHODS: Discrepant test results of sperm acrosin activity (spectrophotometric assay) vs. sperm morphology (strict criteria) in the initial diagnostic investigation of 107 infertile couples were evaluated with respect to fertilization rate (% oocytes with 2 pronuclei) further obtained in IVF treatment. RESULTS: Acrosin activity positively correlated with sperm morphology (% normal forms) (r=0.537) and fertilization rate (r=0.526). ROC curves for the prediction of > or =50% fertilization rate were comparable for acrosin activity and sperm morphology, with optimal cutoff values at 25 microIU/10(6) sperm and 10%, respectively. In multiple regression analysis, sperm acrosin activity (P=0.002) predicted fertilization rate independently of sperm morphology (P<0.001) and sperm vitality (eosin-nigrosin stain) (P=0.03). Acrosin activities > or =25 microIU/10(6) sperm were observed in 36% of severe teratozoospermic samples (< or =4% normal spermatozoa) associated with low fertilization rate. Twenty percent of the morphologically normal ejaculates showed a low acrosin activity (<25 microIU/10(6) sperm) and low hypoosmotic swelling test (HOST) scores (31.4+/-7.6%) and were associated with low fertilization rate. CONCLUSION: The sperm acrosin assay can help to predict sperm fertilizing capacity in IVF independently of sperm morphology.

Oxidized low-density lipoprotein, iron stores, and haptoglobin polymorphism.

BACKGROUND: In vitro experimental studies demonstrated that iron promotes free radical-induced low-density lipoprotein (LDL) oxidation. OBJECTIVE: To test the hypothesis that circulating oxidized LDL (oxLDL) levels might be associated with body iron stores (serum ferritin) and iron-related genetic markers (hemochromatosis gene C282Y mutation, haptoglobin polymorphism). METHODS: We investigated 381 (176 males, 205 females, age 45 +/- 6 years) healthy Caucasians. Serum oxLDL, assayed by a mAb-4E6-based enzyme-linked immunosorbent assay (ELISA), was expressed as oxLDL/LDL ratio to adjust for serum LDL-cholesterol concentration. Hemochromatosis gene C282Y mutation analysis was performed by a Taqman-based polymerase chain reaction (PCR) assay. Haptoglobin (Hp) phenotypes (Hp 1-1, Hp 2-1, Hp 2-2) were determined by starch gel electrophoresis. RESULTS: In stepwise multivariate regression analysis, gender (P < 0.0001), current smoking (P < 0.0001), HDL-cholesterol (P = 0.0001), ferritin (P = 0.0051), body mass index (BMI) (P = 0.0063), and Hp phenotype (P = 0.0331) independently predicted oxLDL/LDL ratio in the total group. In men, smoking (P < 0.0001), ferritin (P = 0.0052), Hp phenotype (P = 0.0063), and HDL-cholesterol (P = 0.0127) were independent determinants of oxLDL/LDL ratio. In women, only body mass index (P < 0.0001), HDL-cholesterol (P = 0.0005), and smoking (P = 0.0025) were significantly associated with oxLDL/LDL ratio. The C282Y mutation (wild-type versus C282Y heterozygotes) was not associated with oxLDL/LDL ratio in both sexes. CONCLUSION: Serum ferritin concentration and Hp polymorphism are independently associated with circulating oxLDL levels in males.

Effect of dialyser membrane pore size on plasma homocysteine levels in haemodialysis patients.

BACKGROUND: Hyperhomocysteinaemia is a putative risk factor for atherothrombotic cardiovascular disease in the haemodialysis population. High-dose vitamin B therapy does not entirely normalize elevated plasma total homocysteine (tHcy) levels in haemodialysis patients. Alternative therapies to reduce tHcy further are therefore required. Modifications of the dialysis regimen may result in a better removal of Hcy. We examined the effect of dialyser membrane pore size on tHcy levels in vitamin-replete chronic haemodialysis patients. METHODS: Forty-five haemodialysis patients were dialysed during 4 weeks with a low-flux, a high-flux and a super-flux membrane, in random order. Pre-dialysis tHcy was determined at baseline and every 4 weeks. In 18 patients, plasma tHcy before and after dialysis and dialysate tHcy concentrations were measured. RESULTS: Pre-dialysis tHcy decreased significantly during 4 weeks super-flux dialysis (-14.6 +/- 2.8%), whereas it remained stable during high-flux (+0.5 +/- 2.4%) and low-flux dialysis (+1.7 +/- 3.2%). The homocysteine reduction ratio was not different for the three membranes: 0.39 +/- 0.03 for the super-flux, 0.47 +/- 0.02 for the high-flux and 0.39 +/- 0.02 for the low-flux dialyser. The amount of Hcy recovered in the dialysate during a single dialysis session was also similar: 117.5 +/- 3.6 micro mol during super-flux, 95.3 +/- 11.5 micro mol during high-flux and 116.5 +/- 11.6 micro mol during low-flux dialysis. CONCLUSION: Super-flux dialysis significantly lowers tHcy in chronic haemodialysis patients. Improved removal of middle-molecule uraemic toxins with inhibitory effects on Hcy-metabolizing enzymes, rather than better dialytic clearance of Hcy itself, may explain the beneficial effect of the super-flux membrane.

Serum zinc in healthy Belgian children.

Many reports mention marginal zinc status in childhood. Information on serum zinc (Zn) in Belgian children since the last reports are old and feeding habits are changing. Four hundred fifty-seven healthy children (0-14 yr, 262 boys) had a venipuncture after an overnight fast during a vaccination campaign. Serum Zn, alpha-tocopherol (alpha-T), cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), apolipoprotein B (Apo B), Apo A, and malondialdehyde (MDA) were determinated. The median Zn value is lower in infants than in older children (respectively 11.6 micromol/L vs 12.8 micromol/L). The type of infant feeding does not influence the serum Zn concentrations (breast-feeding, adapted, hypoallergenic, soy, or thickened). No children had increased serum MDA concentrations and the value is not influenced by the Zn concentration. Children presenting higher serum Zn values also have significantly higher serum alpha-T levels. In infants, there is a significant positive correlation between serum Zn and cholesterol, LDL-C, and Apo B. In this apparently healthy population, no signs of abnormal in vivo peroxidation of fatty acids are observed, even in the children with low serum Zn. More sensitive methods for the detection of peroxidation are necessary for determination of in vivo effects of marginal trace element status.

Polyclonal IgM influence oligodendrocyte precursor cells in mixed glial cell cultures: implications for remyelination.

Polyclonal immunoglobulins for intravenous use (IVIg) are a potent immunomodulator and have been shown to be effective in several immune-mediated diseases. This includes inflammatory demyelinating diseases of the central nervous system (CNS) like multiple sclerosis (MS). Besides their immunomodulatory function, IVIg have been proposed to enhance remyelination based on studies in the animal model of Theiler's murine encephalomyelitis virus (TMEV). Disappointingly, recent treatment trials in patients with MS have failed to demonstrate repair of longstanding deficits. Since the clinical trials have used IVIg that contained nearly exclusively IgG, whereas the most pronounced effect in TMEV was seen with IgM, this could be a possible explanation for the negative outcome in the MS trials. Here we have examined the effects of a new polyclonal IgM preparation (IVIgM) on cultured oligodendrocyte precursor cells (OPCs). To achieve successful remyelination, OPCs proliferate, migrate, and differentiate into mature myelinating oligodendrocytes. IVIgM and commercial IVIg preparations had no influence on proliferation and differentiation of either isolated OPCs or OPCs in coculture with microglia. In contrast, IVIgM inhibited the proliferation of OPCs in mixed glial cultures containing astrocytes and microglia. This was not seen in cultures treated with IVIg, albumin, or interferon-gamma (IFN-gamma), suggesting that this is a specific effect of IVIgM. Differentiation was slightly delayed by IVIgM in mixed glial cultures, but this was not statistically significant and interferon-gamma had a similar effect. These results underline the importance of IgM in influencing OPCs and corroborate the in vivo findings that polyclonal IgM are more potent than IgG in their capacity to influence remyelination. The exact mechanism of how this modulation of OPCs is achieved remains unknown, but a complex interaction among all cells present in the CNS has to be postulated.

The effect of supplementation with an antioxidant preparation on LDL-oxidation is determined by haptoglobin polymorphism.

The genetic polymorphism of haptoglobin (Hp) is an independent risk factor in the pathogenesis of atherosclerosis, a condition in which decreased resistance to in vitro oxidation of LDL-cholesterol is observed. We hypothesised that the Hp polymorphism is one of the factors modulating the resistance to Cu(2+)-induced oxidation of LDL during antioxidant supplementation. In this study, 74 middle-aged subjects with increased oxidative stress were allocated to either matched placebo or oral antioxidative treatment (Quatral) once daily for 16 weeks. Study parameters were increase of lag phase (DeltaLAG) and the ratio of lag phase during treatment period versus baseline (relative oxidation resistance, ROR), measured by Cu(2+)-induced oxidation of isolated LDL. Hp phenotypes were determined by starch gel electrophoresis. A significant and persistent increase of DeltaLAG (P < 0.05) and ROR (P < 0.01) were observed after 16 weeks of active treatment versus placebo. Interindividual differences in both parameters were significantly associated with the Hp polymorphism: in the active treatment group, DeltaLAG and ROR were significantly higher in Hp 1-1 subjects (P < 0.01) compared to Hp 2-1 and 2-2. Our data demonstrate that Hp phenotype is one of the modulating factors determining the increased resistance to Cu(2+)-induced oxidation of LDL during antioxidative treatment.