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Industrial melanism in peppered moths has been studied most intensively in Britain. The first melanic phenotype (effectively solid black) was recorded near Manchester in 1848. By 1895 about 98% of the specimens near Manchester were melanic, and this once rare phenotype had spread across regions of the country blackened by industrial soot. In rural, unpolluted regions, well away from industrial centers, the pale phenotype (peppered with white and black scales) remained the predominant form. During the latter half of the 20th century, following legislation designed to improve air quality, melanics began to decline in frequency and are now rare where once they had been common. Similar evolutionary changes have occurred elsewhere, but records from outside Britain are fragmentary. We have extended previous surveys of American peppered moth populations and present a composite picture of the recent decline in melanism in northern industrial states-Michigan and Pennsylvania-where melanic phenotypes decreased from more than 90% in 1959 to 6% by 2001. We contrast these changes to the near absence of melanism in a southern state-Virginia-during that same period. As in Britain, the decline in melanism in American peppered moths followed clean air legislation.

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Differential cell adhesion, a suggested guiding force for tissue rearrangement during embryogenesis, could be affected by desmosome frequency. A model system for studying embryonic tissue-positioning behaviour involves combining different tissues and following their rearrangements. We have previously shown that for one tissue, embryonic chick heart ventricle, direction of tissue positioning can be altered experimentally. Heart tissue precultured for 2.5 days tends to segregate internally, while tissue pre-cultured for just half a day tends to segregate externally. Also, intact fragments of tissue tend to segregate internally, while reaggregates of trypsin-disaggregated tissues tend to segregate externally. We show here that treatments that increase the tendency to internalize also increase the frequency of adherens junctions and treatments that increase the tendency to externalize decrease the frequency of junctions. An identical hierarchical ordering of the 4 experimental tissues occurs with respect to positioning behaviour and desmosome frequency. In the hierarchy, 2.5-day-cultured fragments greater than 2.5-day-cultured reaggregates greater than 0.5-day-cultured fragments 0.5-day-cultured reaggregates, tissues to the left tend to segregate internally and to have more desmosomes. Tissues to the right segregate externally and have fewer desmosome. This is what is expected if desmosome are organelles for adhesion and if differential adhesion is a factor in tissue-positioning behaviour.

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Disaggregation of seven-day embryonic chick neural retina tissue by eleven different agents produces cells which require variable periods of time to become aggregation competent in culture. The time required between cell isolation and the beginning of inter-cellular adhesion in rotation culture is dependent upon the particular isolating agent used and ranges from zero to sixty minutes.

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Human bone marrow obtained from patients with neutropenia contains a cell population which is absent or diminished in normal marrow. The abnormal population is composed of cells of volume 200-300 mum3 which sediment at 5-5 to 8-5 mm/h. Normal marrow contains one cell class giving rise to increased numbers of grnaulocyte colonies after mass culture, while marrow obtained from neutropenic patients, or from patients with marrow regeneration, shows two such populations; one of these cell classes corresponds to the abnormally large cells demonstrated on velocity sedimentation analysis. This population of large cells may represent a group of either self-renewing cells related to the committed granulocyte progenitors or the pluripotent stem cell.

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Previous studies have indicated that cell sorting and tissue spreading are caused by cell combination-specific differences in intercellular adhesive energies, acting in a system of motile cells. We wished to determine whether these adhesive energies could drive cell rearrangements as well as guide them. Accordingly, aggregates of intermixed embryonic cells were cultured in solutions of the drug cytochalasin B (CCB) at a concentration shown to inhibit the locomotion of cells on a solid surface. In addition, spherical aggregates of several kinds were cultured in mutual contact under similar conditions. Both cell sorting and tissue spreading were found to be inhibited. The prompt release of this inhibition upon removal of the CCB showed that the inhibited cells were not merely injured. Moreover, aggregation experiments showed that CCB did not prevent cells of several kinds from initiating mutual adhesions. In fact, heart cell aggregation was enhanced by CCB. We conclude that interfacial forces, originating outside the cell, act together with forces originating inside it in bringing about the morphogenetic movements of cell sorting and tissue spreading. We propose the term "cooperative cell locomotion" to describe translational movements of cells arising from such a combination of intrinsic and extrinsic forces.

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