Single-pair fluorescence resonance energy transfer on freely diffusing molecules: Observation of Förster distance dependence and subpopulations

Photon bursts from single diffusing donor-acceptor labeled macromolecules were used to measure intramolecular distances and identify subpopulations of freely diffusing macromolecules in a heterogeneous ensemble. By using DNA as a rigid spacer, a series of constructs with varying intramolecular donor-acceptor spacings were used to measure the mean and distribution width of fluorescence resonance energy transfer (FRET) efficiencies as a function of distance. The mean single-pair FRET efficiencies qualitatively follow the distance dependence predicted by Förster theory. Possible contributions to the widths of the FRET efficiency distributions are discussed, and potential applications in the study of biopolymer conformational dynamics are suggested. The ability to measure intramolecular (and intermolecular) distances for single molecules implies the ability to distinguish and monitor subpopulations of molecules in a mixture with different distances or conformational states. This is demonstrated by monitoring substrate and product subpopulations before and after a restriction endonuclease cleavage reaction. Distance measurements at single-molecule resolution also should facilitate the study of complex reactions such as biopolymer folding. To this end, the denaturation of a DNA hairpin was examined by using single-pair FRET.

Differential Distribution of Dynamin Isoforms in Mammalian Cells

Dynamins are 100-kDa GTPases that are essential for clathrin-coated vesicle formation during receptor-mediated endocytosis. To date, three different dynamin genes have been identified, with each gene expressing at least four different alternatively spliced forms. Currently, it is unclear whether these different dynamin gene products perform distinct or redundant cellular functions. Therefore, the focus of this study was to identify additional spliced variants of dynamin from rat tissues and to define the distribution of the dynamin family members in a cultured rat epithelial cell model (Clone 9 cells). After long-distance reverse transcription (RT)-PCR of mRNA from different rat tissues, the full-length cDNAs encoding the different dynamin isoforms were sequenced and revealed four additional spliced variants for dynamin I and nine for dynamin III. Thus, in rat tissues there are a total of at least 25 different mRNAs produced from the three dynamin genes. Subsequently, we generated stably transfected Clone 9 cells expressing full-length cDNAs of six different spliced forms tagged with green fluorescent protein. Confocal or fluorescence microscopy of these transfected cells revealed that many of the dynamin proteins associate with distinct membrane compartments, which include clathrin-coated pits at the plasma membrane and the Golgi apparatus, and several undefined vesicle populations. These results indicate that the dynamin family is more extensive than was originally predicted and suggest that the different dynamin proteins are localized to distinct cytoplasmic or membrane compartments.

Nuclear Pore Complex Number and Distribution throughout the Saccharomyces cerevisiae Cell Cycle by Three-Dimensional Reconstruction from Electron Micrographs of Nuclear Envelopes

The number of nuclear pore complexes (NPCs) in individual nuclei of the yeast Saccharomyces cerevisiae was determined by computer-aided reconstruction of entire nuclei from electron micrographs of serially sectioned cells. Nuclei of 32 haploid cells at various points in the cell cycle were modeled and found to contain between 65 and 182 NPCs. Morphological markers, such as cell shape and nuclear shape, were used to determine the cell cycle stage of the cell being examined. NPC number was correlated with cell cycle stage to reveal that the number of NPCs increases steadily, beginning in G1-phase, suggesting that NPC assembly occurs continuously throughout the cell cycle. However, the accumulation of nuclear envelope observed during the cell cycle, indicated by nuclear surface area, is not continuous at the same rate, such that the density of NPCs per unit area of nuclear envelope peaks in apparent S-phase cells. Analysis of the nuclear envelope reconstructions also revealed no preferred NPC-to-NPC distance. However, NPCs were found in large clusters over regions of the nuclear envelope. Interestingly, clusters of NPCs were most pronounced in early mitotic nuclei and were found to be associated with the spindle pole bodies, but the functional significance of this association is unknown.

An adaptive algorithm for clustering cumulative probability distribution functions using the Kolmogorov–Smirnov two-sample test

This paper proposes an adaptive algorithm for clustering cumulative probability distribution functions (c.p.d.f.) of a continuous random variable, observed in different populations, into the minimum homogeneous clusters, making no parametric assumptions about the c.p.d.f.’s. The distance function for clustering c.p.d.f.’s that is proposed is based on the Kolmogorov–Smirnov two sample statistic. This test is able to detect differences in position, dispersion or shape of the c.p.d.f.’s. In our context, this statistic allows us to cluster the recorded data with a homogeneity criterion based on the whole distribution of each data set, and to decide whether it is necessary to add more clusters or not. In this sense, the proposed algorithm is adaptive as it automatically increases the number of clusters only as necessary; therefore, there is no need to fix in advance the number of clusters. The output of the algorithm are the common c.p.d.f. of all observed data in the cluster (the centroid) and, for each cluster, the Kolmogorov–Smirnov statistic between the centroid and the most distant c.p.d.f. The proposed algorithm has been used for a large data set of solar global irradiation spectra distributions. The results obtained enable to reduce all the information of more than 270,000 c.p.d.f.’s in only 6 different clusters that correspond to 6 different c.p.d.f.’s.

Distribution of basalt boulders and ice rafted debris in the region of the Iceland-Faroe-Ridge (Fig. 6-12)

Psephitic particles in the region of the Iceland-Faeroe-Ridge have been transported and deposited by means of a complex interplay of glacier movements and drifting icebergs. The composition of the particle association is controlled by the sedimentation of basaltic rock particles derived from the ridge itself and, in addition to that and in southern parts of the ridge, from the Faeroe Islands, the Faeroe-Bank and the Bill Baileys-Bank. Besides, there are crystalline and sedimentary dropstones showing a very varied petrography and a wide range of particle sizes. Their percentage becomes greater as the distance from the ridge increases. The association of dropstones is relatively homogeneous in the region of the ridge and only at greater distances from the ridge it becomes more differentiated. Owing to their composition and distribution, as well as on the basis of characteristic fossils and rock types, the drop-stones are derived from Scandianvia and Great Britain. During periods of maximum glaciation, the Icland-Faeroe-Ridge, th eFaeroe-Bank and the Bill Baileys-Bank were under ice.

Spatial distribution of key zooplankton species using continuous plankton recorder (CPR) data from the North Atlantic (2000-2009)

The continuous plankton recorder (CPR) survey is an upper layer plankton monitoring program that has regularly collected samples, at monthly intervals, in the North Atlantic and adjacent seas since 1946. Water from approximately 6 m depth enters the CPR through a small aperture at the front of the sampler and travels down a tunnel where it passes through a silk filtering mesh of 270 µm before exiting at the back of the CPR. The plankton filtered on the silk is analyzed in sections corresponding to 10 nautical miles (approx. 3 m**3 of seawater filtered) and the plankton microscopically identified (Richardson et al., 2006 and reference therein). In the present study we used the CPR data to investigate the current basin scale distribution of C. finmarchicus (C5-C6), C. helgolandicus (C5-C6), C. hyperboreus (C5-C6), Pseudocalanus spp. (C6), Oithona spp. (C1-C6), total Euphausiida, total Thecosomata and the presence/absence of Cnidaria and the Phytoplankton Colour Index (PCI). The PCI, which is a visual assessment of the greenness of the silk, is used as an indicator of the distribution of total phytoplankton biomass across the Atlantic basin (Batten et al., 2003). Monthly data collected between 2000 and 2009 were gridded using the inverse-distance interpolation method, in which the interpolated values were the nodes of a 2 degree by 2 degree grid. The resulting twelve monthly matrices were then averaged within the year and in the case of the zooplankton the data were log-transformed (i.e. log10 (x+1).

Ensemble prediction distribution maps of macroalgae for current conditions and four climate change scenarios and high resultion bathymetry for Potter Cove, WAP, Antarctica

Species distribution models (SDM) predict species occurrence based on statistical relationships with environmental conditions. The R-package biomod2 which includes 10 different SDM techniques and 10 different evaluation methods was used in this study. Macroalgae are the main biomass producers in Potter Cove, King George Island (Isla 25 de Mayo), Antarctica, and they are sensitive to climate change factors such as suspended particulate matter (SPM). Macroalgae presence and absence data were used to test SDMs suitability and, simultaneously, to assess the environmental response of macroalgae as well as to model four scenarios of distribution shifts by varying SPM conditions due to climate change. According to the averaged evaluation scores of Relative Operating Characteristics (ROC) and True scale statistics (TSS) by models, those methods based on a multitude of decision trees such as Random Forest and Classification Tree Analysis, reached the highest predictive power followed by generalized boosted models (GBM) and maximum-entropy approaches (Maxent). The final ensemble model used 135 of 200 calculated models (TSS > 0.7) and identified hard substrate and SPM as the most influencing parameters followed by distance to glacier, total organic carbon (TOC), bathymetry and slope. The climate change scenarios show an invasive reaction of the macroalgae in case of less SPM and a retreat of the macroalgae in case of higher assumed SPM values.

(Table 1) Distribution of Cretaceous-Tertiary benthic foraminifers in ODP Hole 129-802A

Reworked shallow-water larger and deep-water calcareous benthic foraminifers were recovered from foraminiferal packstones and nannofossil chalks in Hole 802A. The autochthonous zeolitic pelagic claystone is characterized by late Campanian abyssal agglutinated foraminifers that allow correlation with the North Atlantic and the adjacent Pigafetta Basin. Assemblages of DendrophryalRhizammina in graded beds within the zeolitic claystone indicate reworking through entrainment in the flocculent E layer of turbidites, rather than recolonization following a biosiliceous event. Background sedimentation of the claystone took place below the carbonate compensation depth. The nannofossil chalk contains reworked lower bathyal to abyssal calcareous foraminifers of late Paleocene to early Miocene age. The topmost bed of the nannofossil chalk unit commences with an algal foraminiferal packstone containing Lepidocyclina sumatrensis, Heterostegina borneensis, Amphistegina hauerina, Asterigerina marshallana, and A. tentoria, which indicate that the source area was a shallow-water reef and allow the bed to be dated as early Miocene. The absence of obviously younger planktonic microfossils in the graded bed indicates that the resedimentation event was generally contemporaneous with original deposition and took place during an early Miocene global sea-level highstand. An early Miocene shallow-water assemblage is also seen in the graded beds at the base of a volcaniclastic turbidite sequence overlying the nannofossil chalks. Resedimentation of this unit was associated with volcanic activity some distance away.

Ice rafted debris distribution in sediment core CRP-1

Lonestone abundances in CRP-1 were investigated using three methods: core examination at Cape Roberts Camp, analysis of digital core images and follow-up core examination. For all images of split-core, we determined size and depth of every detectable lonestone larger than 3 mm. Lonestone abundance decreases exponentially with clast size. Although no significant depth-dependent variations in lonestone size distribution were detected, a strong 0.5-0.7 m abundance periodicity, of unknown origin, is evident within diamicts. Lonestone volume percentage was estimated from size distribution: most size classes contribute approximately the same volume to the total. Sizes >16 mm have rare enough lonestones that their counts are nonrepresentative when based on short intervals of split core. This problem does not affect total counts significantly, but the volume analysis needs to be confined to <= 6 mm lonestones to avoid instability induced by rare and nonrepresentative larger lonestones. If lonestone abundance can be used as an indicator of glacial proximity, then our CRP-1 lonestone abundance logs confirm the overall character of previously inferred variations in relative distance to the ice margin. Large-scale changes in lonestone abundance also reflect the CRP-1 sequence stratigraphy, with individual sequences generally characterised by basal lonestone-rich diamict overlain by lonestone-poor sands and muds. The relationship between glacial proximity and lonestone abundance within diamicts and within sand-mud intervals is, however, less certain. For example, two or three gradual lonestone increases may indicate regressions during glacial advances, in contrast to the more common CRP-l pattern of dominantly transgressive sequences.

An investigation into effects of long-distance seed dispersal on organelle population genetic structure and colonization rate: a model analysis

A simulation-based modelling approach is used to examine the effects of stratified seed dispersal (representing the distribution of the majority of dispersal around the maternal parent and also rare long-distance dispersal) on the genetic structure of maternally inherited genomes and the colonization rate of expanding plant populations. The model is parameterized to approximate postglacial oak colonization in the UK, but is relevant to plant populations that exhibit stratified seed dispersal. The modelling approach considers the colonization of individual plants over a large area (three 500 km x 10 km rolled transects are used to approximate a 500 km x 300 km area). Our approach shows how the interaction of plant population dynamics with stratified dispersal can result in a spatially patchy haplotype structure. We show that while both colonization speeds and the resulting genetic structure are influenced by the characteristics of the dispersal kernel, they are robust to changes in the periodicity of long-distance events, provided the average number of long-distance dispersal events remains constant. We also consider the effects of additional physical and environmental mechanisms on plant colonization. Results show significant changes in genetic structure when the initial colonization of different haplotypes is staggered over time and when a barrier to colonization is introduced. Environmental influences on survivorship and fecundity affect both the genetic structure and the speed of colonization. The importance of these mechanisms in relation to the postglacial spread and genetic structure of oak in the UK is discussed.

"mind the gap": The size-distance dissociation in visual neglect is a cueing effect

There is a growing body of evidence that the processes mediating the allocation of spatial attention within objects may be separable from those governing attentional distribution between objects. In the neglect literature, a related proposal has been made regarding the perception of (within-object) sizes and (between-object) distances. This proposal follows observations that, in size-matching and bisection tasks, neglect is more strongly expressed when patients are required to attend to the sizes of discrete objects than to the (unfilled) distances between objects. These findings are consistent with a partial dissociation between size and distance processing, but a simpler alternative must also be considered. Whilst a neglect patient may fail to explore the full extent of a solid stimulus, the estimation of an unfilled distance requires that both endpoints be inspected before the task can be attempted at all. The attentional cueing implicit in distance estimation tasks might thus account for their superior performance by neglect patients. We report two bisection studies that address this issue. The first confirmed, amongst patients with left visual neglect, a reliable reduction of rightward error for unfilled gap stimuli as compared with solid lines. The second study assessed the cause of this reduction, deconfounding the effects of stimulus type (lines vs. gaps) and attentional cueing, by applying an explicit cueing manipulation to line and gap bisection tasks. Under these matched cueing conditions, all patients performed similarly on line and gap bisection tasks, suggesting that the reduction of neglect typically observed for gap stimuli may be attributable entirely to cueing effects. We found no evidence that a spatial extent, once fully attended, is judged any differently according to whether it is filled or unfilled.

Host-dependent Anopheles flavirostris larval distribution reinforces the risk of malaria near water

Malaria control strategies are more likely to be successful if groups at high risk can be accurately predicted. Given that mosquitoes have an obligate aquatic phase we were interested in determining how vector larval abundance relates to the spatial distribution of human malaria infection. We examined the relationship between malaria parasite prevalence and distance from vector larval habitat, and vector larval abundance and distance from human habitation, in separate studies in rural, low-endemic areas of the Philippines. Parasite prevalence among symptomatic patients was significantly higher among those living in proximity ( less than or equal to 50 m) to potential larval habitats of the major vector, Anopheles flavirostris (adjusted odds ratio [AOR] 2.64, P = 0.02 and AOR 3.43, P = 0.04). A larval survey of A. flavirostris revealed a higher density of early and late instars near human habitation (adjusted P < 0.05). The results suggest that larvae are associated with human habitation, thereby reinforcing malaria risk in people living close to larval habitats. This has implications for understanding the interaction between vectors, hosts, and parasites, and the potential for success of localized malaria control measures.

Correlation between anammox activity and microscale distribution of nitrite in a subtropical mangrove sediment

The distribution of anaerobic ammonium oxidation (anammox) in nature has been addressed by only a few environmental studies, and our understanding of how anammox bacteria compete for substrates in natural environments is therefore limited. In this study, we measure the potential anammox rates in sediment from four locations in a subtropical tidal river system. Porewater profiles of NOx- (NO2- plus NO3-) and NO2- were measured with microscale biosensors, and the availability of NO2- was compared with the potential for anammox activity. The potential rate of anammox increased with increasing distance from the mouth of the river and correlated strongly with the production of nitrite in the sediment and with the average concentration or total pool of nitrite in the suboxic sediment layer. Nitrite accumulated both from nitrification and from NOx- reduction, though NOx- reduction was shown to have the greatest impact on the availability of nitrite in the suboxic sediment layer. This finding suggests that denitrification, though using NO2- as a substrate, also provides a substrate for the anammox process, which has been suggested in previous studies where microscale NO2- profiles were not measured.