Structural analysis of human neutrophil migration: Centriole, microtubule, and microfilament orientation and function during chemotaxis

Orientation of nucleus, centriole, microtubules, and microfilaments within human neutrophils in a gradient of chemoattractant (5 percent Escherichia coli endotoxin-activated serum) was evaluated by electron microscopy. Purified neutropils (hypaque-Ficoll) were placed in the upper compartment of chemotactic chambers. Use of small pore (0.45 μm) micropore filters permitted pseudopod penetration, but impeded migration. Under conditions of chemotaxis with activated serum beneath the filter, the neutrophil population oriented at the filter surface with nuclei located away from the stimulus, centrioles and associated radial array of microtubules beneath the nuclei, and microfilament-rich pseudopods penetrating the filter pores. Reversal of the direction of the gradient of the stimulus (activated serum above cells) resulted in a reorientation of internal structure which preceded pseudopod formation toward the activated serum and migration off the filter. Coordinated orientation of the entire neutrophil population did not occur in buffer (random migration) or in a uniform concentration of activated serum (activated random migration). Conditions of activated random migration resulted in increased numbers of cells with locomotory morphology, i.e. cellular asymmetry with linear alignment of nucleus, centriole, microtubule array, and pseudopods. Thus, activated serum increased the number of neutrophils exhibiting locomotory morphology, and a gradient of activated serum induced the alignment of neutrophils such that this locomotory morphology was uniform in the observed neutrophil populayion. In related studies, cytochalasin B and colchicines were used to explore the role of microfilaments and microtubules in the neutrophil orientation and migration response to activated serum. Cytochalasin B (3.0 μg/ml) prevented migration and decreased the microfilaments seen, but allowed normal orientation of neutrophil structures. In an activated serum gradient, colchicines, but not lumicolchicine, decreased the orientation of nuclei and centrioles, and caused a decrease in centriole-associated microtubules in concentrations as low as 10(-8) to 10(-7) M. These colchicines effects were associated with the rounding of cells and impairment of pseudopod formation. The impaired pseudopod formation was characterized by an inability to form pseudopods in the absence of a solid substrate, a formation of narrow pseudopods within a substrate, and a defect in pseudopod orientation in an activated serum gradient. Functional studies of migration showed that colchicines, but not lumicolchicine, minimally decreased activated random migration and markedly inhibited directed migration, but had not effect on random migration. These studies show that, although functioning microfilaments are probably necessary for neutrophil migration, intact microtubules are essential for normal pseudopod formation and orientation, and maximal unidirectional migration during chemotaxis.     

Worldwide patterns of mitochondrial DNA differentiation in the harbor seal (Phoca vitulina)

The harbor seal (Phoca vitulina) has one of the broadest geographic distributions of any pinniped, stretching from the east Baltic, west across the Atlantic and Pacific Oceans to southern Japan. Although individuals may travel several hundred kilometers on annual feeding migrations, harbor seals are generally believed to be philopatric, returning to the same areas each year to breed. Consequently, seals from different areas are likely to be genetically differentiated, with levels of genetic divergence increasing with distance. Differentiation may also be caused by long-standing topographic barriers such as the polar sea ice. We analyzed samples of 227 harbor seals from 24 localities and defined 34 genotypes based on 435 bp of control region sequence. Phylogenetic analysis and analysis of molecular variance showed that populations in the Atlantic and Pacific Oceans and east and west coast populations of these oceans are significantly differentiated. Within these four regions, populations that are geographically farthest apart generally are the most differentiated and often do not share genotypes or differ in genotype frequency. The average corrected sequence divergence between populations in the Atlantic and Pacific Oceans is 3.28% +/- 0.38% and those among populations within each of these oceans are 0.75% +/- 0.69% and 1.19% +/- 0.65%, respectively. Our results suggest that harbor seals are regionally philopatric, on the scale of several hundred kilometers. However, genetic discontinuities may exist, even between neighboring populations such as those on the Scottish and east English coasts or the east and west Baltic. The mitochondrial data are consistent with an ancient isolation of populations in both oceans, due to the development of polar sea ice. In the Atlantic and Pacific, populations appear to have been colonized from west to east with the European populations showing the most recent common ancestry. We suggest the recent ancestry of European seal populations may reflect recolonization from Ice Age refugia after the last glaciation.      

Complete intra-annual cycle of leaf area index in a Platanus orientalis L. stand

Leaf area index (LAI) analysis of deciduous forest trees is usually restricted to seasonal monitoring involving the assessment of distinct leaf phenological stages within definite time intervals of the year. However, continuous LAI monitoring that includes entire leaf periods is necessary to define the ecophysiological characteristics of deciduous trees. Therefore, this study investigated the intra-annual cycle of the LAI for a Platanus orientalis L. stand in the Bart?n watershed of Turkey. A complete cycle involves three periods: foliation, stable, and defoliation. The foliation period comprises budburst, leaf emergence and flushing sessions, whereas the defoliation period consists of leaf senescence and leaf fall sessions. The stable period is in between these two periods when LAI values are at a climax around maximum. Eight points were determined in the field for the analysis of LAI by a hemispherical photography technique. Over a relatively frequent schedule, photographs were taken almost weekly during the foliation period. Both weekly and approximate monthly photographs were applied during the stable period. Finally, near-monthly photographs were taken for the defoliation period. The foliation period lasted for about 1.5 months from mid-April to May with the mean LAI reaching from 0.16 up to 2.38. Mean LAI was between 2.38 and 2.47 for a stable period over 2 months (June and July). For the defoliation period, mean LAI dropped from 2.42 down to 0.35 over 5 months from August to December. The total foliated period was more than 8 months, which is relatively long for a temperate forest. In addition, correlations between mean LAI and maximum, mean and minimum temperatures were highly significant (P < 0.01) with coefficients (r) of 0.79, 0.90 and 0.93, respectively. By describing the intra-annual LAI pattern, this study fills a gap in the literature on the phenology of Platanus orientalis L.     

The environmental significance of algae in the mi Llandovery succession of the central Oslo Region

Girvanella , micritization phenomena and the dasycladacean alga Cyclocrinites are found in the middle Llandovery succession of the central Oslo Region. Both Girvanella and micritization phenomena occur in carbonate-dominated sequences where sedimentary structures and associated faunas suggest depositional environments above normal wave base. In contrast, Cyclocrinites occurs in shale-dominated sequences where other evidence suggests quiet, deep-water environments. We suggest that Cyclocrinites would not normally be preserved in situ in its life environment in relatively turbulent shallow waters. Preservation most commonly resulted from post mortem transport into low-energy muddy environments devoid of in situ algal activity. The resultant fossil is a flattened cast of this originally spherical alga.     

Quest for wine yeasts—An old story revisited

Numerous studies have described the yeast biota of grapes, and grape must in order to understand better the succession of yeasts during fermentation of wine. The origin of the wine yeasts has been rather controversial. By using more elaborate isolation methods, classical genetic analysis and electrophoretic karyotyping of monosporic clones, with this study, credible proof now exists that the vineyard is the primary source for the wine yeasts and that strains found on the grapes can be followed through the fermentation process.     

Molecular population genetics of the alcohol dehydrogenase locus in the Hawaiian drosophilid D. mimica

Sequence variation among 10 alleles of the alcohol dehydrogenase (Adh) gene of the Hawaiian drosophilid D. mimica was analyzed with reference to the evolutionary history of the Hawaiian subgroup as well as to levels and patterns of polymorphism of the Adh gene in continental drosophilid species. The Adh gene of D. mimica is less polymorphic than that of other drosophilid species, and no replacement substitutions were found. Statistical analyses of the Adh alleles suggested the action of balancing selection and revealed significant linkage disequilibrium among three of the variable sites. The effective population size was estimated to be only slightly smaller than that of continental species and, surprisingly, on the same order of magnitude as the actual size.