RHD allelic identification among D-Brazilian blood donors as a routine test using pools of DNA.

BACKGROUND: RHD alleles leading to a reduced expression of D antigen of the red blood cell (RBC) surface may be erroneously typed as D- by serology and may cause anti-D immunizations when transfused to recipients. METHODS: To determine the occurrence of such alleles among apparent D- blood donors, molecular typing was implemented as a routine test using a pool of DNA. A total of 2,450 pretyped D- samples were tested in pools of 10 for the RHD-specific polymorphism in intron 4 and exon 7. Samples in polymer chain reaction (PCR) positive pools were individually reevaluated by exon-specific PCRs, sequencing, and serologic methods. RESULTS: Among 2,450 serologically D- blood donor samples tested, 101 (4.1%) carried the RHD gene. Nonfunctional RHD (RHDψ, RHD*CE(2-9)-D, and RHD*CE(3-7)-D), different weak D alleles such as RHD*weak D type 1, RHD*weak D type 4.3, RHD*weak D type 5, RHD*weak D type 38, and RHD*DEL were identified. CONCLUSION: We employed a PCR-based assay for RHD as a routine test using pools of ten DNA blood donor samples. The integration of RHD genotyping into the routine screening program using pools of DNA samples was straightforward. As a consequence, 19 (0.8%) blood donors carrying a weak D and Del phenotypes with the potential of causing anti-D immunizations in recipients were reclassified as D+. For each population, it would be necessary to adapt the RHD genotyping strategy to the spectrum of prevalent alleles.

Plasma kinetics of an artificial emulsion resembling chylomicrons in patients with chronic lymphocytic leukemia.

Chylomicrons are the lipoproteins that transport dietary lipids in the blood. Although neoplastic diseases are often accompanied by alterations in lipid metabolism, chylomicrons are scarcely explored in cancer, despite their importance for the body's energy supply. Moreover, no data are available regarding chylomicron metabolism in chronic lymphocytic leukemia (CLL). Chylomicron metabolism in the bloodstream consists of lipolysis by lipoprotein lipase and uptake of remnants by the liver and is difficult to assess in the human body. Among the methods to evaluate this pathway, the determination of the plasma kinetics of triglyceride-rich emulsions that mimic chylomicrons is a practical and straightforward approach. A double-labeled chylomicron-resembling emulsion was injected into 10 patients with CLL and into 11 normolipidemic healthy subjects. The plasma kinetic curves of the emulsion 3H-triglyceride and 14Ccholesteryl ester were determined in plasma samples collected over 30 min. The fractional clearance rate (FCR) of triglycerides in CLL was not changed compared with controls. The FCR of cholesteryl esters was also no different from controls. These results indicate that chylomicron lipolysis and remnant removal are not affected in CLL.