Analytical performance evaluation of a particle-enhanced turbidimetric cystatin C assay on the Roche COBAS 6000 analyzer

ObjectivesCystatin C is a low molecular protein that has been proposed to estimate the glomerular filtration rate. Here we investigated the performance of the Roche cystatin C assay on the COBAS 6000 analyzer.Design and methodsWe studied the imprecision, recovery, limit of detection and quantification, linearity and interferences. For method comparison, split sample aliquots were assayed using the described method and a Siemens cystatin C assay.ResultsThe assay displayed a low total imprecision and a good linearity over the entire range tested. Bilirubin and triglycerides did not interfere with the assay, and only a haemoglobin concentration higher than 6 g/dl interfered with the assay. The assay agreed well with the Siemens assay.ConclusionThe Roche cystatin C assay is an acceptable method for determining the cystatin C and the glomerular filtration rate estimate. On a COBAS 6000, the assay improves and simplifies the laboratory's workload.     

Developmental changes in nerve growth factor (NGF) binding and NGF receptor proteins trkA and p75 in the facial nerve

This study characterizes the temporal-spatial distribution of nerve growth factor (NGF) low (p75) and high-affinity (trkA) receptors in the facial nerve and geniculate ganglion (GG) of developing quail embryos (E-3 to E-14). We used ¹²⁵I-labeled NGF (¹²⁵I-NGF) to study binding dynamics in a temporal series of isolated primordia and an autoradiographic series of staged specimens to characterize the occurrence and distribution of NGF receptors in this cranial nerve and its ganglion. In addition, expression of trkA and p75 protein-like immunoreactivity in the facial nerve and GG was studied by Western blot, in order to distinguish between high- and low-affinity NGF receptors respectively. The quantitative study of binding show that isolated facial primordia ranging from E-3 to E-14 exhibit different levels of specific binding. High initial binding levels were observed on E-3 specimens, then an initial decrease on day 4 (E-4) followed by a steady increase from days E-4 to E-7. Maximum ¹²⁵I-NGF binding was achieved on E-7, followed by a steady decline in binding on days 8 (E-8) and 9 (E-9), reaching near background levels on day 10 (E-10) of development and until the oldest stage assayed (E-14). Most of the cells bearing NGF receptors appeared to be nonneuronal crest-derived cells, but some placode-derived neurons and motor fibers of the VIIth cranial nerve transiently expressed the ability to bind ¹²⁵I-NGF. The temporal pattern of p75 expression matches the pattern of quantitative binding of NGF, while the trkA expression is restricted to a few stages mainly E7 and E9, implying that most of the binding detected is via low-affinity receptors, except for a proportion of high-affinity receptors present at stages of maximum binding. This temporal pattern of NGF binding sites suggests that cells within the VIIth cranial nerve are responsive to and/or dependent upon NGF in vivo, so NGF may play a biological role during normal development of the facial nerve. In view of the developmental events that parallel the occurrence and type of NGF binding sites, we suggest that this role may be to modulate from earlier chemotaxis and cell proliferation to much later events, such as neuronal differentiation and neuron-glia interactions. The significance of these findings in regeneration during adult life remain to be investigated.     

An evaluation of the mode of action framework for mutagenic carcinogens case study II: Chromium (VI)

In response to the 2005 revised U.S Environmental Protection Agency's (EPA) Cancer Guidelines, a strategy is being developed to include all mutagenicity and other genotoxicity data with additional information to determine whether the initiating step in carcinogenesis is through a mutagenic mode of action (MOA). This information is necessary to decide if age‐dependent adjustment factors (ADAFs) should be applied to the risk assessment. Chromium (VI) [Cr (VI)], a carcinogen in animals and humans via inhalation, was reassessed by the National Toxicology Program (NTP) in 2‐year drinking water studies in rodents. From these data, NTP concluded that the results with Cr (VI) showed clear evidence of carcinogenicity in male and female mice and rats. Cr (VI) is also mutagenic, in numerous in vitro assays, in animals (mice and rats) and in humans. Accordingly, Cr (VI) was processed through the MOA framework; postulated key steps in tumor formation were interaction of DNA with Cr (VI) and reduction to Cr (III), mutagenesis, cell proliferation, and tumor formation. Within the timeframe and tumorigenic dose range for early events, genetic changes in mice (single/double‐stranded DNA breaks) commence within 24 hr. Mechanistic evidence was also found for oxidative damage and DNA adduct formation contributing to the tumor response. The weight of evidence supports the plausibility that Cr (VI) may act through a mutagenic MOA. Therefore, the Cancer Guidelines recommend a linear extrapolation for the oral risk assessment. Cr (VI) also induces germ cell mutagenicity and causes DNA deletions in developing embryos; thus, it is recommended that the ADAFs be applied. Environ. Mol. Mutagen., 2010. Published 2009 Wiley‐Liss, Inc.     

Moderate vagal withdrawal in 3.5‐year‐old children is associated with optimal performance on executive function tasks

Vagal tone (measured via respiratory sinus arrhythmia, RSA) and vagal withdrawal (measured by decreases in RSA) have been identified as physiological measures of self‐regulation, but little is known how they may relate to the regulation of cognitive activity as measured through executive function (EF) tasks. We expected that baseline measures of vagal tone, thought to be an indicator of attention, would correlate with EF performance. We also predicted that vagal withdrawal would allow for the reorientation of attention that is needed to succeed on EF tasks, but too much withdrawal would be detrimental. RSA measured at baseline was indeed related to EF performance in 220 3.5‐year‐old children, and those who exhibited a moderate decrease in RSA during the EF tasks outperformed children whose RSA decreased by too little or too much. These findings implicate vagal tone withdrawal as a psychophysiological measure of higher cognitive processes, most likely substantiated through increases in the levels of focused attention. © 2010 Wiley Periodicals, Inc. Dev Psychobiol 52: 603–608, 2010.