The quest for four-dimensional imaging in plant cell biology: it's just a matter of time

BACKGROUND: Analysis of plant cell dynamics over time, or four-dimensional imaging (4-DI), represents a major goal of plant science. The ability to resolve structures in the third dimension within the cell or tissue during developmental events or in response to environmental or experimental stresses (i.e. 4-DI) is critical to our understanding of gene expression, post-expression modulations of macromolecules and sub-cellular system interactions. SCOPE: Microscopy-based technologies have been profoundly integral to this type of investigation, and new and refined microscopy technologies now allow for the visualization of cell dynamics with unprecedented resolution, contrast and experimental versatility. However, certain realities of light and electron microscopy, choice of specimen and specimen preparation techniques limit the scope of readily attaining 4-DI. Today, the plant microscopist must use a combinatorial strategy whereby multiple microscopy-based investigations are used. Modern fluorescence, confocal laser scanning, transmission electron and scanning electron microscopy provide effective conduits for synthesizing data detailing live cell dynamics and highly resolved snapshots of specific cell structures that will ultimately lead to 4-DI. This review provides a synopsis of such technologies available.     

Hox Genes: it's all a matter of context

Recent studies provide compelling new evidence that Hox gene effects depend on fine-structure spatial and temporal information. Further, in a specific cell type, only one or a few downstream genes may mediate Hox morphogenetic functions. If this is generally true, it will have important implications for how Hox regulatory networks operate and evolve.     

Sizing up community genetics: it's a matter of scale

The burgeoning field of community genetics posits that genetic variation within species affects the structure and dynamics of associated communities and ecosystems. While many experiments support this hypothesis, we argue that the most commonly employed experimental design suffers from a fundamental flaw that might result in overestimating the importance of genetic variation. Specifically, most studies collect genotypes from a wide area but perform experiments in small common gardens. Since environmental and genetic variation typically increase with spatial scale, this mismatch in scale is predicted to artificially inflate estimates of the ecological importance of genetic effects. Furthermore, most existing studies have used study systems with particular ecological characteristics, which might further inflate the inferred importance of genetic variation. To critically examine this potential problem, we reanalyze previous studies in community genetics and show how current methods lead to biased conclusions. More specifically, while a growing body of literature shows that intraspecific genetic variation can have an effect, it does not accurately estimate its effect size. As a remedy to this bias, we propose an experimental design that can accurately quantify the importance of genetic and environmental variation in affecting communities and ecosystems.     

Panmixia in the European eel: a matter of time..

The European eel (Anguilla anguilla L.) has been a prime example of the panmixia paradigm because of its extraordinary adaptation to the North Atlantic gyral system, semelparous spawning in the Sargasso Sea and long trans-oceanic migration. Recently, this view was challenged by the suggestion of a genetic structure characterized by an isolation-by-distance (IBD) pattern. This is only likely if spawning subpopulations are spatially and/or temporally separated, followed by non-random larval dispersal. A limitation of previous genetic work on eels is the lack of replication over time to test for temporal stability of genetic structure. Here, we hypothesize that temporal genetic variation plays a significant role in explaining the spatial structure reported earlier for this species. We tested this by increasing the texture of geographical sampling and by including temporal replicates. Overall genetic differentiation among samples was low, highly significant and comparable with earlier studies (FST = 0.0014; p < 0.01). On the other hand, and in sharp contrast with current understandings, hierarchical analyses revealed no significant inter-location genetic heterogeneity and hence no IBD. Instead, genetic variation among temporal samples within sites clearly exceeded the geographical component. Our results provide support for the panmixia hypothesis and emphasize the importance of temporal replication when assessing population structure of marine fish species.     

Concerted evolution of satellite DNA in Sarcocapnos: a matter of time

SarkOne is a genus-specific satellite-DNA family, isolated from the genomes of the species of the genus Sarcocapnos. This satellite DNA is composed of repeats with a consensus length of 855 bp and a mean G+C content of 52.5%. We have sequenced a total of 189 SarkOne monomeric repeats belonging to a total of seven species of the genus Sarcocapnos. The comparative analysis of these sequences both at the intraspecific and the interspecific levels have revealed divergence patterns between species are proportional to between-species divergence according to the phylogeny of the genus. Our study demonstrates that the molecular drive leading to the concerted-evolution pattern of this satellite DNA is a time-dependent process by which new mutations are spreading through genomes and populations at a gradual pace. However, time is a limiting factor in the observation of concerted evolution in some pairwise comparisons. Thus, pairwise comparisons of species sharing a recent common ancestor did not reveal nucleotide sites in transitional stages higher than stage III according to the Strachan's model. By contrast, there was a gradation in the percentage of upper transition stages (IV, V, VI) the more phylogenetically distant the species were. In addition, closely related species shared a high number of polymorphic sites, but these types of sites were not common when comparing more distant species. All these data are discussed in the light of current life-cycle models of satellite-DNA evolution.     

Abuse of residents: it's time to take action.

The scientific study of the sexual dynamics that come into play during residency training seems to both fascinate and repel trainees and their supervisors. One of the more provocative and shameful dimensions of this area of inquiry, the abuse of residents, causes a good deal of distress. How do we respond to findings of significant psychological abuse, discrimination on the basis of sex or sexual orientation and sexual harassment in medical settings? How can we ignore over a decade of research? How can we not heed the experience of so many young physicians? Given the uncertain times in Canadian medicine and the insecurity in our professional and personal lives, we must work together to improve the culture of our teaching institutions and implement measures nationally and locally to close this dark chapter.     

Why it's time for a national health program in the United States.

The United States lacks a coherent national health program. Current programs leave major gaps in coverage and recently have become more restrictive. Influential policies that have failed to correct crucial problems of the health-care system include competitive strategies, corporate intervention, and public-sector cutbacks with bureaucratic expansion. A national health program that combines elements of national health insurance and a national health service is a policy that would help solve current health-care problems. Previous proposals for national health insurance contained weaknesses that would need correction under a national program. Based on the experiences of other economically advanced countries, a national health program could provide universal entitlement to health care while controlling costs and improving the health-care system through structural reorganization. Current proposals for a national health program contain several basic principles dealing with the scope of services, copayments, financing, cost controls, physician and professional associations, personnel and distribution, prevention, and participation in policy making. Support for a national health program is growing rapidly. Such a program would help protect all people who live in this country from unnecessary illness, suffering, and early death.     

Breakdown in classical biological control of Argentine stem weevil: a matter of time

Earlier study at a national scale has shown that insect pests in agriculture can develop resistance to natural enemies following ongoing expansion and simplification of agricultural systems. Here, we used 25 years of field-sampling data segmented into three distinct ecoregions in New Zealand to show that parasitism rate of a key pasture pest (Listronotus bonariensis, Argentine stem weevil) by the introduced parasitoid Microctonus hyperodae has significantly declined. However, this decline has not happened simultaneously in all three ecoregions but with a one year time-lag. The variation in parasitism rate trends might be attributed to subsets of the weevil populations that became resistant to their biocontrol agent once having been exposed to seven years selection pressure (ca. 14 generations) since the parasitoid releases. This result supports the hypothesis that adaptation leading to resistance might have similarly occurred in different parts of the country indicating that the genetic variation needed for the acquisition of resistance was equally present everywhere.     

Replication time of interspersed repetitive DNA sequences in hamsters

The replication time of 34 hamster genomic DNA segments containing interspersed repeat sequences was determined by probing the cloned segments with nick-translated early- and late-replicating hamster DNA. One-third of these cloned families replicated early, one-third replicated late, and one-third replicated without temporal bias. 19 different inserts from these clones along with the SINE, Alu, and the LINE, A36Fc, were used to probe Southern blots of early- and late-replicating hamster or human DNA. We report long interspersed repeats, LINEs, are selectively partitioned into late-replicating DNA and are often concertedly hypomethylated, while short interspersed repeats, SINEs, are selectively partitioned into early-replicating DNA. For some interspersed repeat families, this partitioning is complete or almost complete. The CCGG frequency is very low in late-replicating DNA. The mammalian chromosome's pattern of early-replicating R-bands and late-replicating G-bands reflects a differential distribution of LINEs and SINEs.