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Biomedical subjects

A Manzoni

Publications and source records attributed to A Manzoni.

At least 19 recordsLinked to original sources

Detection of human apolipoprotein E genotypes by DNA biosensors coupled with PCR.

Apolipoprotein E (apoE) is an important constituent of several plasma lipoproteins and has been associated with the risk of developing cardiovascular diseases and in familiar type III hyperlipoproteinemia. We developed new procedures for the detection of apolipoprotein E polymorphism in human blood based on coupling DNA electrochemical or piezoelectric sensors with polymerase chain reaction (PCR). The electrochemical sensor was obtained by immobilizing single-stranded oligonucleotides onto graphite screen-printed electrodes by adsorption at controlled potential. The hybridization reaction that occurred on the electrode surface was evidenced by chronopotentiometric stripping analysis-using daunomycin as indicator. In the piezoelectric sensor, biotinylated 23-mer probes were immobilized on the streptavidin-coated gold surface of a quartz crystal; streptavidin was covalently bound to the thiol/dextran modified gold surface. The hybridization of the immobilized probes with complementary and mismatched DNA was investigated. With the use of two different probes, it was possible to investigate both positions in which apoE polymorphism takes place and consequently, to distinguish the different genotypes. The procedure was validated with both kinds of biosensor with a reference method based on polyacrilamide gel electrophoresis.

Apolipoproteins E↗

Ion effects in measurement of sodium and ionized calcium in direct potentiometry.

In some instruments that measure sodium directly in whole blood, plasma, or serum using ion selective electrodes (direct potentiometry), the higher the ionic strength of the solution, the lower is the sodium recovery in serum, as predicted by theory. The same could be expected for ionized calcium. When measuring the recovery of serum sodium on indirect potentiometric instrument and by flame photometry, which determine concentration in prediluted samples, and on direct potentiometric instruments, we observed that two out of the three direct potentiometry instruments showed a decreased recovery of sodium, as the ionic strength was increased, while on all the other instruments the recovery was complete. No effect of increased ionic strength was noted on the ionized calcium measurements in serum on all the instruments tested. Analysing pure aqueous solutions of sodium and calcium chloride with increased ionic strength on the same instruments, the sodium recovery was always complete or positive, and the same was true for ionized calcium. We postulate some effect of ionic strength on the salt bridge of the measuring systems, which is different when analysing serum or pure aqueous solutions.

Calcium↗

Incomplete recovery of sodium determined in direct potentiometry in blood samples added with heparin salts.

The recent introduction on the market of apparatus for combined blood gas-electrolyte and ionized calcium analyses has raised the problem of whether the same sample can be submitted to all the analyses without generating problems for some analytes. The problem is basically connected to the common use of heparin as an anti-coagulant. To elucidate the possible effect of heparin on measurements we added pooled sera to tubes containing dried heparin/sodium chloride in increasing quantity. Our data demonstrate that, for one direct potentiometric instrument, the sodium recovery is reduced by a moderately high (70-130 IU ml-1) concentration of heparin or by the concomitant addition of the sodium ion which increases the ionic strength of the sample. The effect of the increased sodium chloride was studied separately, adding pooled sera to tubes containing dried sodium chloride in increasing quantity. The sodium recovery was reduced when the ionic strength of the sample was increased. We were unable to separate the negative effects on the sodium recovery due to heparin and due to the increased ionic strength in the sample.

Heparin↗