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Macrophages were then treated with 0.04% solution of trypan blue for 2 min to remove the non-internalized latex beads and cells were finally rinsed with PBS prior to observation under fluorescent microscope (Olympus, Center Valley, PA)

A key contributor to the Caribbeans high mortality is the prevalence of late-stage disease at diagnosis, resulting from the factors outlined earlier

These include a wide range of measures: limiting availability, warning labels, nicotine replacement therapy (NRT) and establishing non-smoking areas (Berridge 1998