PubMed · 5637237
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1968-02-23. The unitest system.. https://pubmed.ncbi.nlm.nih.gov/5637237/
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The lipid hydroperoxide content of isolated, native human plasma lipoproteins, was measured, by the luminol-based chemiluminescent reaction, using a highly sensitive single photon counting instrument. The reaction was specific for lipid hydroperoxides since the signal completely disappeared after treatment with the selenoperoxidase specific for lipidic substrates. In this analytical procedure the whole kinetic of photon emission induced by lipid hydroperoxides and hemin in the presence of luminol is integrated, taking advantage of the mono-exponential fitting of the decay of photon emission. The addition of a detergent to the reaction mixture improved the precision of the measurements apparently by preventing oxidative chain reactions affecting the shape of the decay of photon emission. The sensitivity of the instrument allowed measurements on samples containing just a few picomoles of hydroperoxides, small enough to minimize the effect of antioxidants and quenchers possibly present in the sample (as in the case of lipoproteins). Thus, by using an internal calibration with a phospholipid hydroperoxide, the evaluation of the lipid hydroperoxide content in whole, native lipoproteins was possible without previous extraction and chromatographic separation. Data obtained from plasma lipoproteins isolated by different procedures suggest that lipid hydroperoxide content increases during ultracentrifugation.
The efficacy of sedimentation combined with cytocentrifugation of sputa was examined by preparing smears for the detection of mycobacteria. The detection of acid-fast bacilli from sputum sediment by cytocentrifugation yielded 32 smears positive for acid-fast bacilli, out of which in 26 cases better results were obtained than by the conventional method. More bacilli were found in 20 cases and on 6 occasions solely the cytospin method was positive. The differences in the number of bacilli were 100-200-fold in 5 cases and 10-fold in 12 paired cases. The technique applied is suitable for making culture and smear from the same sediment. The sedimentation combined with cytocentrifugation increases the effectiveness of the detection of acid-fast bacilli in smears.