Persistent Cell-Autonomous Circadian Oscillations in Fibroblasts Revealed by Six-Week Single-Cell Imaging of PER2::LUC Bioluminescence

Biological oscillators naturally exhibit stochastic fluctuations in period and amplitude due to the random nature of molecular reactions. Accurately measuring the precision of noisy oscillators and the heterogeneity in period and strength of rhythmicity across a population of cells requires single-cell recordings of sufficient length to fully represent the variability of oscillations. We found persistent, independent circadian oscillations of clock gene expression in 6-week-long bioluminescence recordings of 80 primary fibroblast cells dissociated from PER2::LUC mice and kept in vitro for 6 months. Due to the stochastic nature of rhythmicity, the proportion of cells appearing rhythmic increases with the length of interval examined, with 100% of cells found to be rhythmic when using 3-week windows. Mean period and amplitude are remarkably stable throughout the 6-week recordings, with precision improving over time. For individual cells, precision of period and amplitude are correlated with cell size and rhythm amplitude, but not with period, and period exhibits much less cycle-to-cycle variability (CV 7.3%) than does amplitude (CV 37%). The time series are long enough to distinguish stochastic fluctuations within each cell from differences among cells, and we conclude that the cells do exhibit significant heterogeneity in period and strength of rhythmicity, which we measure using a novel statistical metric. Furthermore, stochastic modeling suggests that these single-cell clocks operate near a Hopf bifurcation, such that intrinsic noise enhances the oscillations by minimizing period variability and sustaining amplitude

Heterogeneity of soil nutrients and subsurface biota in a dryland ecosystem

Dryland ecosystems have long been considered to have a highly heterogeneous distribution of nutrients and soil biota, with greater concentrations of both in soils under plants relative to interspace soils. We examined the distribution of soil resources in two plant communities (dominated by either the shrub Coleogyne ramosissima or the grass Stipa hymenoides) at two locations. Interspace soils were covered either by early successional biological soil crusts (BSCs) or by later successional BSCs (dominated by nitrogen (N)-fixing cyanobacteria and lichens). For each of the 8 plant type×crust type×locations, we sampled the stem, dripline, and 3 interspace distances around each of 3 plants. Soil analyses revealed that only available potassium (Kav) and ammonium concentrations were consistently greater under plants (7 of 8 sites and 6 of 8 sites, respectively). Nitrate and iron (Fe) were greater under plants at 4 sites, while all other nutrients were greater under plants at less than 50% of the sites. In contrast, calcium, copper, clay, phosphorus (P), and zinc were often greater in the interspace than under the plants. Soil microbial biomass was always greater under the plant compared to the interspace. The community composition of N-fixing bacteria was highly variable, with no distinguishable patterns among microsites. Bacterivorous nematodes and rotifers were consistently more abundant under plants (8 and 7 sites, respectively), and fungivorous and omnivorous nematodes were greater under plants at 5 of the 8 sites. Abundance of other soil biota was greater under plants at less than 50% of the sites, but highly correlated with the availability of N, P, Kav, and Fe. Unlike other ecosystems, the soil biota was only infrequently correlated with organic matter. Lack of plant-driven heterogeneity in soils of this ecosystem is likely due to (1) interspace soils covered with BSCs, (2) little incorporation of above-ground plant litter into soils, and/or (3) root deployment patterns

Accelerated Partial Breast Irradiation (APBI): A review of available techniques

Breast conservation therapy (BCT) is the procedure of choice for the management of the early stage breast cancer. However, its utilization has not been maximized because of logistics issues associated with the protracted treatment involved with the radiation treatment. Accelerated Partial Breast Irradiation (APBI) is an approach that treats only the lumpectomy bed plus a 1-2 cm margin, rather than the whole breast. Hence because of the small volume of irradiation a higher dose can be delivered in a shorter period of time. There has been growing interest for APBI and various approaches have been developed under phase I-III clinical studies; these include multicatheter interstitial brachytherapy, balloon catheter brachytherapy, conformal external beam radiation therapy and intra-operative radiation therapy (IORT). Balloon-based brachytherapy approaches include Mammosite, Axxent electronic brachytherapy and Contura, Hybrid brachytherapy devices include SAVI and ClearPath. This paper reviews the different techniques, identifying the weaknesses and strength of each approach and proposes a direction for future research and development. It is evident that APBI will play a role in the management of a selected group of early breast cancer. However, the relative role of the different techniques is yet to be clearly identified

Response and resilience of soil biocrust bacterial communities to chronic physical disturbance in arid shrublands

The impact of 10 years of annual foot trampling on soil biocrusts was examined in replicated field experiments at three cold desert sites of the Colorado Plateau, USA. Trampling detrimentally impacted lichens and mosses, and the keystone cyanobacterium, Microcoleus vaginatus, resulting in increased soil erosion and reduced C and N concentrations in surface soils. Trampled biocrusts contained approximately half as much extractable DNA and 20–52% less chlorophyll a when compared with intact biocrusts at each site. Two of the three sites also showed a decline in scytonemin-containing, diazotrophic cyanobacteria in trampled biocrusts. 16S rRNA gene sequence and terminal restriction fragment length polymorphism (T-RFLP) analyses of soil bacteria from untrampled and trampled biocrusts demonstrated a reduced proportion (23–65% reduction) of M. vaginatus and other Cyanobacteria in trampled plots. In parallel, other soil bacterial species that are natural residents of biocrusts, specifically members of the Actinobacteria, Chloroflexi and Bacteroidetes, became more readily detected in trampled than in untrampled biocrusts. Replicate 16S rRNA T-RFLP profiles from trampled biocrusts at all three sites contained significantly more fragments (n=17) than those of untrampled biocrusts (n6) and exhibited much higher variability among field replicates, indicating transition to an unstable disturbed state. Despite the dramatic negative impacts of trampling on biocrust physical structure and composition, M. vaginatus could still be detected in surface soils after 10 years of annual trampling, suggesting the potential for biocrust re-formation over time. Physical damage of biocrusts, in concert with changing temperature and precipitation patterns, has potential to alter performance of dryland ecosystems for decades