Kappa free light chains in cerebrospinal fluid as markers of intrathecal immunoglobulin synthesis.
Abstract Background: Intrathecal immunoglobulin synthesis is observed in several inflammatory disorders of the central nervous system, but its detection by current laboratory tests is either tedious or relatively insensitive. We assessed the diagnostic accuracy of an assay for κ free light chains (κFLC) in cerebrospinal fluid (CSF) and serum, and compared it with traditional tests for intrathecal immunoglobulin synthesis. Methods: κFLCs were measured by nephelometry in CSF/serum pairs from 112 patients. Samples were excluded if blood contamination of CSF as a result of traumatic lumbar puncture (n = 12) or monoclonal bands in both CSF and serum (n = 5) were present. The remaining sample pai…
Following ionic activity by electrochemistry during the polymerase chain reaction
The most commonly used technique for gene detection is the polymerase chain reaction (PCR). PCR is associated with alterations in ionic activity because inorganic pyrophosphate (PPi) and inorganic phosphate (Pi) ions are produced during nucleotide polymerization. To maintain electro-neutrality, magnesium, potassium, and ammonium ions are bound to DNA. Deoxynucleotides are also bound to DNA during PCR. Some authors have described DNA itself as an electrically conducting polymer formed by base stapling with the formation of extensive Pi systems. In the current study, alterations in electrical conductivity determined experimentally during PCR are reported, and a model explaining the observed c…
In vitro model for the activation of CD4 and CD8 T cell receptors.
Previously, most models that sought to explain the misregulation of immune cell function assumed molecular similarities between the disease-causing pathogens and the host's proteins. In recent time several different models have been proposed and in this study, these concepts are compared to a new hypothesis proposing another explanation for this immune dysregulation: the possibility that the mislocalization of proteins may be responsible for autoimmune activity. Based on this hypothesis, proteins are recognized as self or non-self depending on where they appear in sufficiently high concentrations. To examine this new idea, the intracellular human proteins beta-actin, GAPDH, and hemoglobin a…
Quantification of PCR products by phosphate measurement
Various techniques for quantification of PCR are available. Most frequently, the densitometric intensities of ethidium bromide-stained PCR products separated in gels are compared after normalizing to the levels of housekeeping gene products such as beta-actin. More precise, but extremely time consuming, is the technique of competitive PCR. Newer methods, such as tracking amplification in real-time, have high start-up and maintenance costs (e.g., TaqMan, Applied Biosystems; LightCycler, Roche; I-Cycler, Bio-Rad). Here, I describe an alternative, simple technique to quantify PCR products by determining the entire phosphate released during PCR. The method can be performed using common laborato…
Activation of T‐Lymphocytes by LDL‐Cholesterol
Native LDL-cholesterol can be mechanically stressed by strong vortexing. According to one hypothesis, mechanical shear stress within the vessel can lead to an aggregation of LDL-cholesterol and subsequently to activation of CD4 and CD8 T-lymphocytes. The goal of this study was to determine the proportion of activated CD4 and CD8 T-lymphocytes that is induced by adding unstressed and mechanically stressed LDL-cholesterol to whole blood samples. Whole blood was taken from 12 healthy subjects. All probands fasted for at least 12 h before blood withdrawal. In each case, 1 ml of whole blood from each subject was incubated for 16 h at 32 degrees C (89.3 degrees F) with concanavalin A (A), without…