A one-tube quantitative HIV-1 RNA NASBA nucleic acid amplification assay using electrochemiluminescent (ECL) labelled probes

Quantification of HIV-1 viral RNA based on co-amplification of an internal standard Q-RNA dilution series requires a number of NASBA nucleic acid amplification reactions. For each internal standard Q-RNA concentration a separate NASBA amplification has to be performed. The development of an electrochemiluminescent (ECL) detection system with a dynamic signal detection range over 5 orders of magnitude enabled simplification of the Q-NASBA protocol. Three distinguishable Q-RNAs (QA, QB and QC) are mixed with the wild-type sample at different amounts (i.e. 10(4) QA, 10(3) QB and 10(2) QC molecules) and co-amplified with the wild-type RNA in one tube. Using ECL-labelled oligonucleotides the wild-type, QA, QB and QC amplificates are separately detected with a semi-automated ECL detection instrument and the ratio of the signals determined. The amount of initial wild-type RNA can be calculated from the ratio of wild-type signal to QA, QB and QC signals. This one-tube Q-NASBA protocol was compared to the earlier described protocol with six amplifications per quantification using model systems and HIV-1 RNA isolated from plasma of HIV-1-infected individuals. In all cases the quantification results of HIV-1 RNA were comparable between the two methods tested. Due to the use of only one amplification per quantification in the one-tube Q-NASBA protocol the QA, QB and QC internal standard RNA molecules can be added to the sample before nucleic acid isolation. The ratio of QA:QB:QC:WT RNAs, from which the initial input of WT-RNA is calculated, will remain constant independent of any loss that might occur during the nucleic acid isolation.     

A portable program to present courseware on microcomputers

In computer assisted instruction, certain main functions which have to be carried out by the computer can he distinguished. The performance of these functions will have different software and hardware requirements in different educational environments and both the requirements and the technology to implement them will change over time. It is therefore advantageous to make CAI systems as adaptable as possible. One approach to this is to divide the systems into independent modules each designed to achieve good portability both for software and for hardware. This paper describes such a module which is part of the Modular CAI System Delft. The program makes it possible to present on different types of microcomputers courseware designed using other modules of the system. The program is implemented in Pascal to yield maximum portability on modern microcomputers. Its future and portability are discussed.     

Acute cardiovascular toxicity of thallium (I) ions

The paper deals with: 1. the protein concentration in the perilymph (PL), the serum and the cerebrospinal fluid (CSF), 2. the protein pattern in the PL and 3. histological findings in the middle and inner ear in unilaterally ear-infected guinea pigs. The studies were performed 6 h to 21 days post infectionem (Fig. 1). The pathological changes in the middle ear, which, in most cases, were limited to the infected ear, were initially evaluated under the operating microscope and divided into 4 stages. The analytical and histological results were presented as functions of these stages. As the inflammation intensity increased, the protein concentration in the PL of the infected ears increased to a level exceeding that of the normal value more than ten times (Fig. 2). However, in the serum and in the CSF this concentration remained unchanged. Likewise, no significant protein increase in the PL of the contralateral ears was detectable in most cases. As the inflammation intensity increased, the number of the precipitation lines detectable immunoelectrophoretically increased in the PL of the infected ears (Fig. 3). An increase in the alpha1- and gamma-globulins and a decrease in Albumin was found by electrophoresis on cellulose acetate strips (Tab. 3). The histological findings correlated with initially established inflammatory stages of the middle ear mucous membrane (Tab. 4). As the inflammation intensity increased, the round window, too, was changed pathologically, so that in some cases of purulent otitis media middle ear secretion could enter the cochlea. The protein increase in the PL immediately after the infection is probably due to an increase in the blood vessel permeability in the inner ear.     

Numerical calculations of the capacitance of linearly graded Si p-n junctions

Exact calculations are made of the capacitance of linearly graded Si p-n junctions, taking the influence of electrons and holes into account. The results agree well with the experimental data for gradients of less than 1022cm?4. The discrepancies for gradients larger than 1022cm?4 are probably due an interface-state mechanism.     

A low noise heterodyne receiver for astronomical observations operating around 0.63 mm wavelength

A heterodyne receiver is described in which an InSb hot electron bolometer is used as a mixer. The local oscillator power is obtained by doubling the frequency of a backward wave oscillator. The receiver operates between 460 and 500 GHz (0.65–0.6 mm). Noise temperatures amount typically to 1000 K.