ABSTRACT
Accurate prognostic evaluation of patients with multiple myeloma (MM) is required for their stratification for more adequate therapy. Chromosomal G-banding and interphase fluorescence in situ hybridization (FISH) on cell-nonspecific samples and on myeloma cells selected by magnetic-activated cell separation (MACS) were used to study 13 samples from 12 multiple myeloma (MM) patients. Bone marrow (BM) samples were analysed using three approaches. Standard mitotic samples were prepared and analysed after G-banding. Interphase FISH was performed to detect the 13q14 deletion in unselected BM cells. In parallel, myeloma cells were selected from the BM using the CD138-specific antibody. The high-purity myeloma cell suspension was then analysed by interphase FISH for the 13q14 deletion. Magnetic separation yielded enriched myeloma cell suspensions with the mean viability of 98.0% (range: 97.0%-99.0%), and the purity of 97.6% (range: 87.2%-99.2%) as detected morphologically, and 85.2% (range: 44.8%-98.4%) as detected by immunophenotyping for CD138+ cells. Interphase FISH revealed the 13q14.3 deletion in 5 of 13 (38.5%) of cell-nonspecific samples and in 9 of 13 (69.2%) of enriched myeloma cell suspensions. In conclusion, interphase FISH on immunomagnetically selected MM cells increases the detection of the 13q14 deletion in BM samples from the patients with MM.
Subject(s)
Chromosome Aberrations , Chromosomes, Human, Pair 13 , Immunomagnetic Separation , Multiple Myeloma/genetics , Cells, Cultured , Chromosome Banding , Gene Deletion , Humans , Immunophenotyping , In Situ Hybridization, Fluorescence , Magnetics , Membrane Glycoproteins/biosynthesis , Mitosis , Multiple Myeloma/blood , Proteoglycans/biosynthesis , Syndecan-1 , SyndecansABSTRACT
Monoterpenes (alpha- and beta-pinene, delta-3-carene, camphene, alpha-phellandrene and limonene) were determined by Gas Chromatography-Mass Spectrometry in fresh needles of Picea abies (L) Karst. situated in three ecologically different regions of Moravia. Through the use of cryogenic grinding for critical sample homogenisation, solvent extraction with cold n-hexane, followed by GC analysis with mass detection, very low quantities of sample (0.1-0.3 g needles) could be processed, thus permitting a comparison of amounts of monoterpenes in needles of different ages and a determination of changes in concentrations of monoterpenes in needles at different locations on the tree. The amount of alpha-phellandrene decreased with the age of the needles, and the content of delta-3-carene was higher in apical branches compared to lateral ones.
