Electric field exposure triggers and guides formation of pseudopod-like blebs in U937 monocytes.

We describe a new phenomenon of anodotropic pseudopod-like blebbing in U937 cells stimulated by nanosecond pulsed electric field (nsPEF). In contrast to "regular," round-shaped blebs, which are often seen in response to cell damage, pseudopod-like blebs (PLBs) formed as longitudinal membrane protrusions toward anode. PLB length could exceed the cell diameter in 2 min of exposure to 60-ns, 10-kV/cm pulses delivered at 10-20 Hz. Both PLBs and round-shaped nsPEF-induced blebs could be efficiently inhibited by partial isosmotic replacement of bath NaCl for a larger solute (sucrose), thereby pointing to the colloid-osmotic water uptake as the principal driving force for bleb formation. In contrast to round-shaped blebs, PLBs retracted within several minutes after exposure. Cells treated with 1 nM of the actin polymerization blocker cytochalasin D were unable to form PLBs and instead produced stationary, spherical blebs with no elongation or retraction capacity. Live cell fluorescent actin tagging showed that during elongation actin promptly entered the PLB interior, forming bleb cortex and scaffold, which was not seen in stationary blebs. Overall, PLB formation was governed by both passive (physicochemical) effects of membrane permeabilization and active cytoskeleton assembly in the living cell. To a certain extent, PLB mimics the membrane extension in the process of cell migration and can be employed as a nonchemical model for studies of cytomechanics, membrane-cytoskeleton interaction and cell motility.

Significance of transmission electron microscopy in subtyping of monocytic leukemia.

The objective of this paper is to produce an ultrastructural classification of acute monocytic leukemia (AML-M5) in relation to clinical behaviors. The ultrastructural characteristics of blasts of the monocytic series were analyzed in 72 M5 patients by transmission electron microscopy (TEM), in terms of their content of typical monoblasts, atypical monoblasts, atypical promonocytes, and typical promonocytes in bone-marrow aspirates. Four kinds of monocytic blasts were identified by cell size and shape, nuclear profile, nucleocytoplasmic ratio, heterochromatin content, nucleolus, granules, vesicles, and Golgi apparatus. Their characteristics of remission rate, cytochemistry, immunophenotype, and cytogenetics were also investigated. The data obtained permitted M5 patients to be divided into monoblast and promonocyte types. Monoblast-type patients expressed weaker monocytic enzymograms and specific antigen staining for CD14 and CD64, compared with promonocyte-type patients. Monoblast patients had higher CR than promonocyte patients. Therefore, TEM subclassification of patients differs from and improves upon the light microscopical criteria for distinguishing monoblasts and promonocytes and has clinical significance.